Long-Term Outcomes of Pharmacoinvasive Strategy Versus Primary Percutaneous Coronary Intervention in ST-Elevation Myocardial Infarction: A Study from Mexico City.

Although primary percutaneous coronary intervention (pPCI) is the treatment of choice in ST-elevation myocardial infarction (STEMI), challenges may arise in accessing this intervention for certain geodemographic groups. Pharmacoinvasive strategy (PIs) has demonstrated comparable outcomes when delays in pPCI are anticipated, but real-world data on long-term outcomes are limited. The aim of the present study was to compare long-term outcomes among real-world patients with STEMI who underwent either PIs or pPCI. This was a prospective registry including patients with STEMI who received reperfusion during the first 12 hours from symptom onset. The primary objective was cardiovascular mortality at 12 months according to the reperfusion strategy (pPCI vs PIs) and major cardiovascular events (cardiogenic shock, recurrent myocardial infarction, and congestive heart failure), and Bleeding Academic Research Consortium type 3 to 5 bleeding events were also evaluated. A total of 799 patients with STEMI were included; 49.1% underwent pPCI and 50.9% received PIs. Patients in the PIs group presented with more heart failure on admission (Killip-Kimbal >I 48.1 vs 39.7, p = 0.02) and had a lower proportion of pre-existing heart failure (0.2% vs 1.8%, p = 0.02) and atrial fibrillation (0.25% vs 1.2%, p = 0.02). No statistically significant difference was observed in cardiovascular mortality at the 12-month follow-up (hazard ratio for PIs 0.74, 95% confidence interval 0.42 to 1.30, log-rank p = 0.30) according to the reperfusion strategy used. The composite of major cardiovascular events (hazard ratio for PIs 0.98, 95% confidence interval 0.75 to 1.29, p = 0.92) and Bleeding Academic Research Consortium type 3 to 5 bleeding rates were also comparable. A low socioeconomic status, Killip-Kimball >2, age >60 years, and admission creatinine >2.0 mg/100 ml were predictors of the composite end point after multivariate analysis. In conclusion, this prospective real-world registry provides additional support that long-term major cardiovascular outcomes and bleeding are not different between patients who underwent PIs versus primary PCI.

Circulating tumour cell enumeration, biomarker analyses, and kinetics in patients with colorectal cancer and other GI malignancies.

Objective: Most of the work in terms of liquid biopsies in patients with solid tumors is focused on circulating tumor DNA (ctDNA). Our aim was to evaluate the feasibility of using circulating tumor cells (CTCs) in peripheral blood samples from patients with advanced or metastatic gastrointestinal (GI) cancers. Methods: In this prospective study, blood samples were collected from each patient in 2 AccuCyte® blood collection tubes and each tube underwent CTC analysis performed utilizing the RareCyte® platform. The results from both tubes were averaged and a total of 150 draws were done, with 281 unique reported results. The cadence of sampling was based on convenience sampling and piggybacked onto days of actual clinical follow-ups and treatment visits. The CTC results were correlated with patient- and tumor-related variables. Results: Data from a total of 59 unique patients were included in this study. Patients had a median age of 58 years, with males representing 69% of the study population. More than 57% had received treatment prior to taking blood samples. The type of GI malignancy varied, with more than half the patients having colorectal cancer (CRC, 54%) followed by esophageal/gastric cancer (17%). The least common cancer was cholangiocarcinoma (9%). The greatest number of CTCs were found in patients with colorectal cancer (Mean: 15.8 per 7.5 ml; Median: 7.5 per 7.5 ml). In comparison, patients with pancreatic cancer (PC) had considerably fewer CTCs (Mean: 4.2 per 7.5 ml; Median: 3 per 7.5 ml). Additionally, we found that patients receiving treatment had significantly fewer CTCs than patients who were not receiving treatment (Median 2.7 versus 0.7). CTC numbers showed noteworthy disparities between patients with responding/stable disease in comparison to those with untreated/progressive disease (Median of 2.7 versus 0). When CTCs were present, biomarker analyses of the four markers human epidermal growth factor receptor 2 (HER2)/programmed death-ligand 1 (PD-L1)/Kiel 67 (Ki-67)/epidermal growth factor receptor (EGFR) was feasible. Single cell sequencing confirmed the tumor of origin. Conclusion: Our study is one of the first prospective real-time studies evaluating CTCs in patients with GI malignancies. While ctDNA-based analyses are more common in clinical trials and practice, CTC analysis provides complementary information from a liquid biopsy perspective that is of value and worthy of continued research.

Analysis of immortal-time effect in post-infarction ventricular septal defect.

Introduction: Time-fixed analyses have traditionally been utilized to examine outcomes in post-infarction ventricular septal defect (VSD). The aims of this study were to: (1) analyze the relationship between VSD closure/non-closure and mortality; (2) assess the presence of immortal-time bias. Material and methods: In this retrospective cohort study, patients with ST-elevation myocardial infarction (STEMI) complicated by VSD. Time-fixed and time-dependent Cox regression methodologies were employed. Results: The study included 80 patients: surgical closure (n = 26), transcatheter closure (n = 20), or conservative management alone (n = 34). At presentation, patients without VSD closure exhibited high-risk clinical characteristics, had the shortest median time intervals from STEMI onset to VSD development (4.0, 4.0, and 2.0 days, respectively; P = 0.03) and from STEMI symptom onset to hospital arrival (6.0, 5.0, and 0.8 days, respectively; P < 0.0001). The median time from STEMI onset to closure was 22.0 days (P = 0.14). In-hospital mortality rate was higher among patients who did not undergo defect closure (50%, 35%, and 88.2%, respectively; P < 0.0001). Closure of the defect using a fixed-time method was associated with lower in-hospital mortality (HR = 0.13, 95% CI 0.05-0.31, P < 0.0001, and HR 0.13, 95% CI 0.04-0.36, P < 0.0001, for surgery and transcatheter closure, respectively). However, when employing a time-varying method, this association was not observed (HR = 0.95, 95% CI 0.45-1.98, P = 0.90, and HR 0.88, 95% CI 0.41-1.87, P = 0.74, for surgery and transcatheter closure, respectively). These findings suggest the presence of an immortal-time bias. Conclusions: This study highlights that using a fixed-time analytic approach in post-infarction VSD can result in immortal-time bias. Researchers should consider employing time-dependent methodologies.

Induced Systemic Resistance in the Bacillus spp.-Capsicum chinense Jacq.-PepGMV Interaction, Elicited by Defense-Related Gene Expression.

Induced systemic resistance (ISR) is a mechanism involved in the plant defense response against pathogens. Certain members of the Bacillus genus are able to promote the ISR by maintaining a healthy photosynthetic apparatus, which prepares the plant for future stress situations. The goal of the present study was to analyze the effect of the inoculation of Bacillus on the expression of genes involved in plant responses to pathogens, as a part of the ISR, during the interaction of Capsicum chinense infected with PepGMV. The effects of the inoculation of the Bacillus strains in pepper plants infected with PepGMV were evaluated by observing the accumulation of viral DNA and the visible symptoms of pepper plants during a time-course experiment in greenhouse and in in vitro experiments. The relative expression of the defense genes CcNPR1, CcPR10, and CcCOI1 were also evaluated. The results showed that the plants inoculated with Bacillus subtilis K47, Bacillus cereus K46, and Bacillus sp. M9 had a reduction in the PepGMV viral titer, and the symptoms in these plants were less severe compared to the plants infected with PepGMV and non-inoculated with Bacillus. Additionally, an increase in the transcript levels of CcNPR1, CcPR10, and CcCOI1 was observed in plants inoculated with Bacillus strains. Our results suggest that the inoculation of Bacillus strains interferes with the viral replication, through the increase in the transcription of pathogenesis-related genes, which is reflected in a lowered plant symptomatology and an improved yield in the greenhouse, regardless of PepGMV infection status.

Accurate isolation and detection of circulating tumor cells using enrichment-free multiparametric high resolution imaging.

Introduction: The reliable and accurate detection of rare circulating tumor cells (CTCs) from cancer patient blood samples promises advantages in both research and clinical applications. Numerous CTC detection methods have been explored that rely on either the physical properties of CTCs such as density, size, charge, and/or their antigen expression profiles. Multiple factors can influence CTC recovery including blood processing method and time to processing. This study aimed to examine the accuracy and sensitivity of an enrichment-free method of isolating leukocytes (AccuCyte® system) followed by immunofluorescence staining and high-resolution imaging (CyteFinder® instrument) to detect CTCs. Method: Healthy human blood samples, spiked with cancer cells from cancer cell lines, as well as blood samples obtained from 4 subjects diagnosed with cancer (2 pancreatic, 1 thyroid, and 1 small cell lung) were processed using the AccuCyte-CyteFinder system to assess recovery rate, accuracy, and reliability over a range of processing times. Results: The AccuCyte-CyteFinder system was highly accurate (95.0%) at identifying cancer cells in spiked-in samples (in 7.5 mL of blood), even at low spiked-in numbers of 5 cells with high sensitivity (90%). The AccuCyte-CyteFinder recovery rate (90.9%) was significantly higher compared to recovery rates obtained by density gradient centrifugation (20.0%) and red blood cell lysis (52.0%). Reliable and comparable recovery was observed in spiked-in samples and in clinical blood samples processed up to 72 hours post-collection. Reviewer analysis of images from spiked-in and clinical samples resulted in high concordance (R-squared value of 0.998 and 0.984 respectively). Discussion: The AccuCyte-CyteFinder system is as an accurate, sensitive, and clinically practical method to detect and enumerate cancer cells. This system addresses some of the practical logistical challenges in incorporating CTCs as part of routine clinical care. This could facilitate the clinical use of CTCs in guiding precision, personalized medicine.

Chagas Heart Disease: Beyond a Single Complication, from Asymptomatic Disease to Heart Failure.

Chagas cardiomyopathy (CC), caused by the protozoan Trypanosoma cruzi, is an important cause of cardiovascular morbidity and mortality in developing countries. It is estimated that 6 to 7 million people worldwide are infected, and it is predicted that it will be responsible for 200,000 deaths by 2025. The World Health Organization (WHO) considers Chagas disease (CD) as a Neglected Tropical Disease (NTD), which must be acknowledged and detected in time, as it remains a clinical and diagnostic challenge in both endemic and non-endemic regions and at different levels of care. The literature on CC was analyzed by searching different databases (Medline, Cochrane Central, EMBASE, PubMed, Google Scholar, EBSCO) from 1968 until October 2022. Multicenter and bioinformatics trials, systematic and bibliographic reviews, international guidelines, and clinical cases were included. The reference lists of the included papers were checked. No linguistic restrictions or study designs were applied. This review is intended to address the current incidence and prevalence of CD and to identify the main pathogenic mechanisms, clinical presentation, and diagnosis of CC.

Liquid Biopsy Assessment of Circulating Tumor Cell PD-L1 and IRF-1 Expression in Patients with Advanced Solid Tumors Receiving Immune Checkpoint Inhibitor.

BACKGROUND: Reliable biomarkers that can be serially monitored to predict treatment response to immune checkpoint inhibitors (ICIs) are still an unmet need. Here, we present a multiplex immunofluorescence (IF) assay that simultaneously detects circulating tumor cells (CTCs) and assesses CTC expression of programmed death ligand-1 (PD-L1) and interferon regulatory factor 1 (IRF-1) as a candidate biomarker related to ICI use. OBJECTIVE: To assess the potential of CTC PD-L1 and IRF-1 expression as candidate biomarkers for patients with advanced epithelial solid tumors receiving ICIs. PATIENTS AND METHODS: We tested the IF CTC assay in a pilot study of 28 patients with advanced solid tumors who were starting ICI. Blood for CTC evaluation was obtained prior to starting ICI, after a single cycle of therapy, and at the time of radiographic assessment or treatment discontinuation. RESULTS: At baseline, patients with 0-1 CTCs had longer progression-free survival (PFS) compared to patients with ≥ 2 CTCs (4.3 vs 1.3 months, p = 0.01). The presence of any PD-L1+ CTCs after a single dose of ICI portended shorter PFS compared to patients with no CTCs or PD-L1- CTCs (1.2 vs 4.2 months, p = 0.02); the presence of any PD-L1+ or IRF-1+ CTCs at time of imaging assessment or treatment discontinuation also was associated with shorter PFS (1.9 vs 5.5 months, p < 0.01; 1.6 vs 4.7 months, p = 0.05). CTC PD-L1 and IRF-1 expression did not correlate with tumor tissue PD-L1 or IRF-1 expression. Strong IRF-1 expression in tumor tissue was associated with durable (≥ 1 year) radiographic response (p = 0.02). CONCLUSIONS: Based on these results, CTC PD-L1 and IRF-1 expression is of interest in identifying ICI resistance and warrants further study.

Single Circulating-Tumor-Cell-Targeted Sequencing to Identify Somatic Variants in Liquid Biopsies in Non-Small-Cell Lung Cancer Patients.

Non-small-cell lung cancer (NSCLC) accounts for most cancer-related deaths worldwide. Liquid biopsy by a blood draw to detect circulating tumor cells (CTCs) is a tool for molecular profiling of cancer using single-cell and next-generation sequencing (NGS) technologies. The aim of the study was to identify somatic variants in single CTCs isolated from NSCLC patients by targeted NGS. Thirty-one subjects (20 NSCLC patients, 11 smokers without cancer) were enrolled for blood draws (7.5 mL). CTCs were identified by immunofluorescence, individually retrieved, and DNA-extracted. Targeted NGS was performed to detect somatic variants (single-nucleotide variants (SNVs) and insertions/deletions (Indels)) across 65 oncogenes and tumor suppressor genes. Cancer-associated variants were classified using OncoKB database. NSCLC patients had significantly higher CTC counts than control smokers (p = 0.0132; Mann-Whitney test). Analyzing 23 CTCs and 13 white blood cells across seven patients revealed a total of 644 somatic variants that occurred in all CTCs within the same subject, ranging from 1 to 137 per patient. The highest number of variants detected in ≥1 CTC within a patient was 441. A total of 18/65 (27.7%) genes were highly mutated. Mutations with oncogenic impact were identified in functional domains of seven oncogenes/tumor suppressor genes (NF1, PTCH1, TP53, SMARCB1, SMAD4, KRAS, and ERBB2). Single CTC-targeted NGS detects heterogeneous and shared mutational signatures within and between NSCLC patients. CTC single-cell genomics have potential for integration in NSCLC precision oncology.

Clinical Validation of a Circulating Tumor Cell Assay Using Density Centrifugation and Automated Immunofluorescence Microscopy.

OBJECTIVES: The US Food and Drug Administration (FDA)-approved CELLSEARCH assay (Menarini Silicon Biosystems) for circulating tumor cells (CTCs) relies on expression of an epithelial cell adhesion molecule to enrich for CTCs. We sought to validate a CTC assay (RareCyte) for clinical use that instead collects a buffy coat preparation enriched for CTCs. METHODS: Normal peripheral blood specimens spiked with cultured breast and prostate cancer cells and 47 clinical samples were used to validate assay performance. Specimens were enriched for buffy coat cells and applied onto 8 glass slides. The slides were immunofluorescently stained and imaged by automated microscopy and computer-aided image analysis. RESULTS: The assay was 100% specific for detecting spiked tumor cells. For samples spiked with 25, 50, and 125 cells, the percentage coefficients of variation were 42%, 21%, and 3.7%, respectively. Linearity studies demonstrated a slope of 0.99, an intercept of 1.6, and R2 of 0.96. Recoveries at the 25-, 50-, and 125-cell levels were 92%, 111%, and 100%, respectively. Clinical samples run on both CELLSEARCH and RareCyte correlated with an R2 of 0.8 after log-transformation and demonstrated 87.5% concordance using the CELLSEARCH criteria for predicting adverse outcomes. CONCLUSIONS: The RareCyte CTC assay has comparable performance to the FDA-cleared method and is ready for further clinical validation studies.

Beyond Circulating Tumor Cell Enumeration: Cell-Based Liquid Biopsy to Assess Protein Biomarkers and Cancer Genomics Using the RareCyte® Platform.

The practice of medicine has steadily employed less invasive methods to obtain information derived from the tumor to guide clinical management of patients. Liquid biopsy-the sampling of blood-is a non-invasive method for generating information previously only available from tissue biopsies of the tumor mass. Analysis of fragmented circulating tumor DNA in the plasma is clinically used to identify actionable mutations and detect residual or recurrent disease. Plasma analysis cannot, however, assess cancer phenotypes, including the expression of drug targets and protein biomarkers. Circulating tumor cells (CTCs) are intact cancer cells that have entered the blood that have the potential for distant metastasis. While enumeration of CTCs is prognostic of outcome, recently developed technology allows for the interrogation of protein biomarkers on CTCs that could be predictive of response. Furthermore, since CTCs contain intact whole cancer genomes, isolating viable CTCs detected during therapy could provide a rational approach to assessing mutational profiles of resistance. Identification, characterization and molecular analysis of CTCs together will advance the capacity of liquid biopsy to meet the requirements of twenty-first century medicine.

Safety, Feasibility, and Merits of Longitudinal Molecular Testing of Multiple Metastatic Sites to Inform mTNBC Patient Treatment in the Intensive Trial of Omics in Cancer.

PURPOSE: Patients with metastatic triple-negative breast cancer (mTNBC) have poor outcomes. The Intensive Trial of Omics in Cancer (ITOMIC) sought to determine the feasibility and potential efficacy of informing treatment decisions through multiple biopsies of mTNBC deposits longitudinally over time, accompanied by analysis using a distributed network of experts. METHODS: Thirty-one subjects were enrolled and 432 postenrollment biopsies performed (clinical and study-directed) of which 332 were study-directed. Molecular profiling included whole-genome sequencing or whole-exome sequencing, cancer-associated gene panel sequencing, RNA-sequencing, and immunohistochemistry. To afford time for analysis, subjects were initially treated with cisplatin (19 subjects), or another treatment they had not received previously. The results were discussed at a multi-institutional ITOMIC Tumor Board, and a report transmitted to the subject's oncologist who arrived at the final treatment decision in conjunction with the subject. Assistance was provided to access treatments that were predicted to be effective. RESULTS: Multiple biopsies in single settings and over time were safe, and comprehensive analysis was feasible. Two subjects were found to have lung cancer, one had carcinoma of unknown primary site, tumor samples from three subjects were estrogen receptor-positive and from two others, human epidermal growth factor receptor 2-positive. Two subjects withdrew. Thirty-four of 112 recommended treatments were accessed using approved drugs, clinical trials, and single-patient investigational new drugs. After excluding the three subjects with nonbreast cancers and the two subjects who withdrew, 22 of 26 subjects (84.6%) received at least one ITOMIC Tumor Board-recommended treatment. CONCLUSION: Further exploration of this approach in patients with mTNBC is merited.

Healthy Photosynthetic Mechanism Suggests ISR Elicited by Bacillus spp. in Capsicum chinense Plants Infected with PepGMV.

The aim of this study was to evaluate the effect of inoculation with Bacillus spp. isolates on the photosynthetic apparatus of Capsicum chinense plants infected with PepGMV. In vitro and greenhouse experiments were performed to evaluate whether the inoculation improved plants' performance through the increase in photosynthetic efficiency to control PepGMV. The results showed that despite PepGMV infection, the plants inoculated with some isolates of Bacillus spp. had a healthy photosynthetic mechanism, as the photochemical parameters and gas exchange increased. The maximum photochemical quantum yield of PSII (Fv/Fm) of plants with PepGMV and inoculated with Bacillus isolates (M9, K46, and K47) increased (7.85, 7.09, and 7.77%, respectively) with respect to uninoculated controls. In inoculated plants, the CO2 assimilation rate increased and the transpiration rate decreased, therefore indicating an increased water use efficiency. This effect was reflected by the less severe symptoms caused by PepGMV in the plants obtained from seeds inoculated with different Bacillus spp. Plants inoculated with K47 isolates showed an increase in fruit yield and quality. This study suggests that it is possible to protect, at the greenhouse level, C. chinense plants from PepGMV through selected rhizobacteria inoculation.

Circulating tumor cell investigation in breast cancer patient-derived xenograft models by automated immunofluorescence staining, image acquisition, and single cell retrieval and analysis.

BACKGROUND: Breast cancer patient-derived xenograft (BC-PDX) models represent a continuous and reproducible source of circulating tumor cells (CTCs) for studying their role in tumor biology and metastasis. We have previously shown the utility of BC-PDX models in the study of CTCs by immunohistochemistry (IHC) on serial paraffin sections and manual microscopic identification of cytokeratin-positive cells, a method that is both low-throughput and labor-intensive. We therefore aimed to identify and characterize CTCs from small volume mouse blood samples and examined its practical workflow in a study of BC-PDX mice treated with chemotherapy using an automated imaging platform, the AccuCyte®-CyteFinder® system. METHODS: CTC analysis was conducted using blood from non-tumor bearing SCID/Beige mice spiked with human breast cancer cells, BC-PDX-bearing mice, and BC-PDX mice treated with vehicle or chemotherapeutic agent(s). After red blood cell lysis, nucleated cells were mixed with transfer solution, processed onto microscope slides, and stained by immunofluorescence. The CyteFinder automated scanning microscope was used to identify CTCs, defined as nucleated cells that were human cytokeratin-positive, and mouse CD45-negative. Disaggregated primary BC-PDX tumors and lung metastatic nodules were processed using the same immunostaining protocol. Collective expression of breast cancer cell surface markers (EpCAM, EGFR, and HER2) using a cocktail of target-specific antibodies was assessed. CTCs and disaggregated tumor cells were individually retrieved from slides using the CytePicker® module for sequence analysis of a BC-PDX tumor-specific PIK3CA mutation. RESULTS: The recovery rate of human cancer cells spiked into murine blood was 83 ± 12%. CTC detection was not significantly different from the IHC method. One-third of CTCs did not stain positive for cell surface markers. A PIK3CA T1035A mutation present in a BC-PDX tumor was confirmed in isolated single CTCs and cells from dissociated metastatic nodules after whole genome amplification and sequencing. CTC evaluation could be simply implemented into a preclinical PDX therapeutic study setting with substantial improvements in workflow over the IHC method. CONCLUSIONS: Analysis of small volume blood samples from BC-PDX-bearing mice using the AccuCyte-CyteFinder system allows investigation of the role of CTCs in tumor biology and metastasis independent of surface marker expression.

Inhibitory effects of quinoid redox mediators on a denitrifying culture.

Denitrification with p-cresol as the electron source was studied in the presence of three quinones at different molar concentrations (0-2 mM): menadione (MEN), alizarine (ALZ) and anthraquinone-2,6-disulfonate (AQDS). Results showed that denitrifying yields were not altered (0.9), but the substrates' consumption efficiency was mainly affected when adding MEN. In the presence of ALZ and MEN, specific consumption rates decreased 40% for p-cresol and 90% for nitrate. The sludge previously exposed to quinones was submitted to recovering denitrifying studies using acetate and p-cresol. After exposing to AQDS and ALZ, the metabolic activity of denitrifying sludge was completely recovered. The previous exposition to any MEN concentration resulted in a very low metabolic recuperation. The results show that MEN and ALZ have a marked inhibitory effect on substrates' consumption and the AQDS does not affect at all. The evidence suggests that MEN modifies the transport system of substrates and ALZ forms a complex with molybdenum. A model based on oxido-reduction potentials of the enzymes involved points out that the influence of quinones tested appears to be more associated with quinone moiety structural properties and hydrophobicity.

Implications of electric potentials applied on a denitrifying process.

The effect of three electric potentials (EP) (+104, -187 and -279 mV) applied to the denitrifying process was explored. It was observed that the denitrifying sludge was able to support the oxidation of p-cresol with the application of the EP in the absence of nitrate, but it was unable to drive the denitrification without an organic electron donor. On denitrification, the applied EP uncoupled the oxidative from the reductive process, favoring the p-cresol oxidation over the production of N2. Additionally, biochemical level effects were observed. At +104 and -279 mV potentials, the nitrate and nitrite consumption was affected as well as the p-hydroxybenzoate transformation. However, at -187 mV, effects seemed to occur only on the transport of substrates. This paper presents evidence that denitrification has very characteristic and different physiological behaviors for each EP assayed.

Presence of Multidrug-Resistant Shiga Toxin-Producing Escherichia coli, Enteropathogenic Escherichia coli, and Enterotoxigenic Escherichia coli on Fresh Cheeses from Local Retail Markets in Mexico.

Cheesemaking is one of the most important industries in Mexico. Among all the Mexican cheeses, fresh cheeses are the most popular and most consumed cheese in Mexico and Latin America. However, in Mexico fresh cheese is frequently made with unpasteurized milk and sold in public markets. This may increase the risk for contamination of dairy products with pathogenic bacteria. The presence of multidrug-resistant pathogenic bacteria in food is an important public health concern. Diarrheagenic Escherichia coli pathotypes (DEPs) are foodborne bacteria. This study investigated the presence of indicator bacteria and multidrug-resistant DEPs in fresh cheeses. A total of 120 fresh cheese samples were collected from public markets in the city of Pachuca, Mexico. The samples were analyzed for presence of fecal coliforms (FC), E. coli, and antibiotic resistant DEPs. FC and E. coli were analyzed using the most-probable-number technique. DEPs were identified using two multiplex PCR methods. Susceptibility to 16 antibiotics was tested for the isolated DEPs strains by the standard assay. The frequency of FC, E. coli, and DEPs in the cheese samples was 50, 40, and 19%, respectively. The identified DEPs included Shiga toxin-producing E. coli (STEC; 8%), enteropathogenic E. coli (EPEC; 6%), and enterotoxigenic E. coli (ETEC; 5%). All isolated strains exhibited resistance to at least five antibiotics. One, one, two, and three STEC strains were resistant to 14, 12, 11, and 10 antibiotics, respectively. One strain of EPEC was resistant to 11 antibiotics, three EPEC strains to 9, and one strain to 7. One, one, and two strains of ETEC were resistant to 10, 8, and 7 antibiotics, respectively. The results of the present study indicate that fresh cheeses made with unpasteurized milk could be a risk for consumers, both for native people and visitors to Mexico.

The RareCyte® platform for next-generation analysis of circulating tumor cells.

Circulating tumor cells (CTCs) can reliably be identified in cancer patients and are associated with clinical outcome. Next-generation "liquid biopsy" technologies will expand CTC diagnostic investigation to include phenotypic characterization and single-cell molecular analysis. We describe here a rare cell analysis platform designed to comprehensively collect and identify CTCs, enable multi-parameter assessment of individual CTCs, and retrieve single cells for molecular analysis. The platform has the following four integrated components: 1) density-based separation of the CTC-containing blood fraction and sample deposition onto microscope slides; 2) automated multiparameter fluorescence staining; 3) image scanning, analysis, and review; and 4) mechanical CTC retrieval. The open platform utilizes six fluorescence channels, of which four channels are used to identify CTC and two channels are available for investigational biomarkers; a prototype assay that allows three investigational biomarker channels has been developed. Single-cell retrieval from fixed slides is compatible with whole genome amplification methods for genomic analysis. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

Behavior of 11 Foodborne Bacteria on Whole and Cut Mangoes var. Ataulfo and Kent and Antibacterial Activities of Hibiscus sabdariffa Extracts and Chemical Sanitizers Directly onto Mangoes Contaminated with Foodborne Bacteria.

The behavior of foodborne bacteria on whole and cut mangoes and the antibacterial effect of Hibiscus sabdariffa calyx extracts and chemical sanitizers against foodborne bacteria on contaminated mangoes were investigated. Mangoes var. Ataulfo and Kent were used in the study. Mangoes were inoculated with Listeria monocytogenes, Shigella flexneri, Salmonella Typhimurium, Salmonella Typhi, Salmonella Montevideo, Escherichia coli strains (O157:H7, non-O157:H7 Shiga toxin-producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative). The antibacterial effect of five roselle calyx extracts (water, ethanol, methanol, acetone, and ethyl acetate), sodium hypochlorite, colloidal silver, and acetic acid against foodborne bacteria were evaluated on contaminated mangoes. The dry extracts obtained with ethanol, methanol, acetone, and ethyl acetate were analyzed by nuclear magnetic resonance spectroscopy to determine solvent residues. Separately, contaminated whole mangoes were immersed in five hibiscus extracts and in sanitizers for 5 min. All foodborne bacteria attached to mangoes. After 20 days at 25 ± 2°C, all foodborne bacterial strains on whole Ataulfo mangoes had decreased by approximately 2.5 log, and on Kent mangoes by approximately 2 log; at 3 ± 2°C, they had decreased to approximately 1.9 and 1.5 log, respectively, on Ataulfo and Kent. All foodborne bacterial strains grew on cut mangoes at 25 ± 2°C; however, at 3 ± 2°C, bacterial growth was inhibited. Residual solvents were not detected in any of the dry extracts by nuclear magnetic resonance. Acetonic, ethanolic, and methanolic roselle calyx extracts caused a greater reduction in concentration (2 to 2.6 log CFU/g) of all foodborne bacteria on contaminated whole mangoes than the sodium hypochlorite, colloidal silver, and acetic acid. Dry roselle calyx extracts may be a potentially useful addition to disinfection procedures of mangoes.

Population fluctuation of Schistocerca piceifrons piceifrons (Orthoptera: Acrididae) in the Yucatán Península and its relation with the environmental conditions / Fluctuación poblacional de Schistocerca piceifrons piceifrons (Orthoptera: Acrididae) en la Península de Yucatán y su relación con las condiciones climáticas

Abstract The Central American Locust Schistocerca piceifrons piceifrons is one of the most damaging plant pest in Mexico and Central America. The present work was carried out to evaluate the seasonal population fluctuation of S. p. piceifrons and vegetation diversity and their association with weather factors and edaphic conditions in the gregarious zone of the Yucatán Península. The study was performed in seven sites during three seasons: North-wind (December 2013), rainy (June 2014) and dry (April 2014). The locust density was sampled in transect of 100 m2, as well as the vegetation in 16 m2: plant species richness (PSR) and relative species density (RSD), and analyzed by generalized linear models. Additionally, soil samples were obtained at 10 cm depth into a 4 × 6 m quadrat, land use in the sites was classified and temperature, precipitation and evaporation of each site were obtained from the database and they were analyzed with multiple factor analysis. The population density of S. p. piceifrons was higher in the sites Panaba, Tizimin, Tunkas and Cenotillo (F= 74.3, P < 0.0001). Characterization of vegetation showed that PSR and RSD were higher during the rainy season relative to those in the dry season (F= 50.4, P < 0.0001). RSD was identified as the most important group associated with locust density (0.86), followed by isotherm/isohyets (0.63), maximum precipitation and temperature (0.60), as well as the land use (0.65); no correlation between locust density and soil characteristics was found. Locust density was positively correlated with the abundance of the grass Panicum maximum (Sr2= 0.85, PC5= 0.87). This work shows that the population of S. p. piceifrons is high in the rainy season and influenced primarily by the abundance of the grass P. maximum and the precipitation. The results indicate that surveys for early detection and control of the locust on the Yucatán Península can focus on areas with the grass P. maximum to predict risk areas and target survey efforts. Rev. Biol. Trop. 66(1): 403-414. Epub 2018 March 01.

Resumen La langosta Centroamericana Schistocerca piceifrons piceifrons es una de las plagas más dañinas en México y Centroamérica. El presente trabajo se realizó para evaluar la fluctuación estacional de poblaciones de S. p. piceifrons y su asociación con la diversidad de vegetación, factores climáticos y edáficos en la zona gregarígena de la Península de Yucatán. Se seleccionaron siete sitios y se muestrearon a lo largo de tres estaciones: nortes (diciembre 2013), lluvias (junio 2014) y sequías (abril 2014). La densidad poblacional de la langosta fue muestreada en transectos de 100 m2, así como la vegetación en 16 m2 obteniendo la riqueza de especies vegetales (REV) y la densidad relativa de especies (DRE), y fueron analizadas por modelos lineales generalizados. Adicionalmente se obtuvieron muestras de suelo de 10 cm de profundidad en un área de 4 × 6 m, se clasificó el uso del suelo de cada sitio y se obtuvieron en una base de datos las condiciones de temperatura, precipitación y evaporación para cada sitio, estos datos se analizaron con análisis de factores múltiples. La densidad poblacional de S. p. piceifrons fue mayor en los sitios de Panaba, Tizimin, Tunkas y Cenotillo (F= 74.3, P < 0.0001). La caracterización de la vegetación mostró que la REV y la DRE fueron mayores en la estación de lluvias que en la de sequías (F= 50.4, P < 0.0001). La DRE fue el grupo más importante asociado a la densidad de la langosta (0.86), seguido por las isotermas/isoyetas (0.63) y la precipitación-temperatura máxima (0.60), así como el uso del suelo (0.65); no hubo correlación con las características del suelo. La densidad de la langosta fue correlacionada positivamente con la abundancia del pasto Panicum maximum (Sr2= 0.85, PC5= 0.87). El estudio mostró que las poblaciones de S. p. piceifrons fueron mayores en la estación lluviosa e influenciadas principalmente por la abundancia del pasto P. maximum y la precipitación. Los resultados indican que las exploraciones, la detección temprana y el control de la langosta en la Península de Yucatán puede enfocarse sobre áreas cultivadas con P. maximum para predecir áreas con riesgo y eficientar los recursos.

RareCyte® CTC Analysis Step 1: AccuCyte® Sample Preparation for the Comprehensive Recovery of Nucleated Cells from Whole Blood.

The RareCyte platform addresses important technology limitations of current circulating tumor cell (CTC) collection methods, and expands CTC interrogation to include advanced phenotypic characterization and single-cell molecular analysis. In this respect, it represents the "next generation" of cell-based liquid biopsy technologies. In order to identify and analyze CTCs, RareCyte has developed an integrated sample preparation, imaging and individual cell retrieval process. The first step in the process, AccuCyte®, allows the separation, collection, and transfer to a slide the nucleated cell fraction of the blood that contains CTCs. Separation and collection are based on cell density-rather than size or surface molecular expression-and are performed within a closed system, without wash or lysis steps, enabling high CTC recovery. Here, we describe our technique for nucleated cell collection from a blood sample, and the spreading of these nucleated cells onto glass slides permitting immunofluorescent staining, cell identification, and individual cell picking described in subsequent chapters. In addition to collection of rare cells such as CTCs, AccuCyte also collects cells of the circulating immune system onto archivable slides as well as plasma from the same sample.