Cutting Boards: An Overlooked Source of Microplastics in Human Food?

Plastic cutting boards are a potentially significant source of microplastics in human food. Thus, we investigated the impact of chopping styles and board materials on microplastics released during chopping. As chopping progressed, the effects of chopping styles on microplastic release became evident. The mass and number of microplastics released from polypropylene chopping boards were greater than polyethylene by 5-60% and 14-71%, respectively. Chopping on polyethylene boards was associated with a greater release of microplastics with a vegetable (i.e., carrots) than chopping without carrots. Microplastics showed a broad, bottom-skewed normal distribution, dominated by <100 µm spherical-shaped microplastics. Based on our assumptions, we estimated a per-person annual exposure of 7.4-50.7 g of microplastics from a polyethylene chopping board and 49.5 g of microplastics from a polypropylene chopping board. We further estimated that a person could be exposed to 14.5 to 71.9 million polyethylene microplastics annually, compared to 79.4 million polypropylene microplastics from chopping boards. The preliminary toxicity study of the polyethylene microplastics did not show adverse effects on the viability of mouse fibroblast cells for 72 h. This study identifies plastic chopping boards as a substantial source of microplastics in human food, which requires careful attention.

Prevalence and Trends of Weakness Among Middle-Aged and Older Adults in the United States.

ABSTRACT: McGrath, R, FitzSimmons, S, Andrew, S, Black, K, Bradley, A, Christensen, BK, Collins, K, Klawitter, L, Kieser, J, Langford, M, Orr, M, and Hackney, KJ. Prevalence and trends of weakness among middle-aged and older adults in the United States. J Strength Cond Res 37(12): 2484-2490, 2023-Muscle weakness, which is often determined with low handgrip strength (HGS), is associated with several adverse health conditions; however, the prevalence and trends of weakness in the United States is not well-understood. We sought to estimate the prevalence and trends of weakness in Americans aged at least 50 years. The total unweighted analytic sample included 22,895 Americans from the 2006-2016 waves of the Health and Retirement Study. Handgrip strength was measured with a handgrip dynamometer. Men with weakness were below at least one of the absolute or normalized (body mass, body mass index) cut points: <35.5 kg, <0.45 kg/kg, <1.05 kg/kg/m 2 . The presence of any weakness in women was also identified as being below one of the absolute or normalized HGS cut points: <20.0 kg, <0.34 kg/kg, or <0.79 kg/kg/m 2 . There was an increasing trend in the prevalence of any weakness over time ( p < 0.001). The prevalence of weakness was 45.1% (95% confidence interval [CI]: 44.0-46.0) in the 2006-2008 waves and 52.6% (CI: 51.5-53.7) in the 2014-2016 waves. Weakness prevalence was higher for older (≥65 years) Americans (64.2%; CI: 62.8-65.5) compared with middle-aged (50-64 years) Americans (42.2%; CI: 40.6-43.8) in the 2014-2016 waves. Moreover, the prevalence of weakness in the 2014-2016 waves was generally higher in women (54.5%; CI: 53.1-55.9) than in men (50.4%; CI: 48.7-52.0). Differences existed in weakness prevalence across races and ethnicities. The findings from our investigation suggest that the prevalence of weakness is overall pronounced and increasing in Americans. Efforts for mitigating and better operationalizing weakness will elevate in importance as our older American population grows.

pH-Sensitive Nanodrug Carriers for Codelivery of ERK Inhibitor and Gemcitabine Enhance the Inhibition of Tumor Growth in Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC), a metabolic disorder, remains one of the leading cancer mortality sources worldwide. An initial response to treatments, such as gemcitabine (GEM), is often followed by emergent resistance reflecting an urgent need for alternate therapies. The PDAC resistance to GEM could be due to ERK1/2 activity. However, successful ERKi therapy is hindered due to low ligand efficiency, poor drug delivery, and toxicity. In this study, to overcome these limitations, we have designed pH-responsive nanoparticles (pHNPs) with a size range of 100-150 nm for the simultaneous delivery of ERKi (SCH 772984) and GEM with tolerable doses. These pHNPs are polyethylene glycol (PEG)-containing amphiphilic polycarbonate block copolymers with tertiary amine side chains. They are systemically stable and capable of improving in vitro and in vivo drug delivery at the cellular environment's acidic pH. The functional analysis indicates that the nanomolar doses of ERKi or GEM significantly decreased the 50% growth inhibition (IC50) of PDAC cells when encapsulated in pHNPs compared to free drugs. The combination of ERKi with GEM displayed a synergistic inhibitory effect. Unexpectedly, we uncover that the minimum effective dose of ERKi significantly promotes GEM activities on PDAC cells. Furthermore, we found that pHNP-encapsulated combination therapy of ERKi with GEM was superior to unencapsulated combination drug therapy. Our findings, thus, reveal a simple, yet efficient, drug delivery approach to overcome the limitations of ERKi for clinical applications and present a new model of sensitization of GEM by ERKi with no or minimal toxicity.

A systematic review of refillable e-liquid nicotine content accuracy.

OBJECTIVES: The use of e-cigarettes is becoming more common in the United States. E-cigarettes are often refilled with nicotine-containing solutions of various concentrations purchased in local shops or on the Internet. There is evidence that the nicotine content in these solutions is often mislabeled; thus, we reviewed the available literature on this topic. DATA SOURCES: We conducted a systematic review of peer-reviewed articles published worldwide on e-liquid nicotine content accuracy using the databases CAB Direct, Cochrane Central Register of Controlled Trials, PubMed, and SPORTDiscus (EBSCO). STUDY SELECTION: Initial screening of titles and abstracts was conducted to determine relevancy for inclusion. Full-article reviews of studies involving the purchase and chemical analysis of nicotine content in refillable e-liquids were conducted for final inclusion. DATA EXTRACTION: Data extraction included e-liquid sample size, whether the samples were labeled to contain nicotine, whether the samples were purchased in retail shops or online, and the number and percentage of samples where the analyzed nicotine content fell outside 10% of the labeled nicotine content. RESULTS: Twenty articles described cross-sectional studies of purchased samples containing nicotine. The number of nicotine-containing e-liquid samples obtained in each study varied from 2 to 71. The percentage of samples with an analyzed nicotine concentration of more than 10% above or below the labeled nicotine concentration ranged from 0% to 100% (277/574 or 48.3%; median 46.85%). A large percentage of the samples deviated by 10% from the labeled nicotine concentrations in both U.S. and non-U.S. samples, with U.S. samples having a higher percentage. CONCLUSION: Our review shows that actual nicotine concentrations in e-liquids may vary considerably from labeled concentrations. Pharmacists should warn patients to be wary of the contents of e-cigarettes, and explain the dangers of using these products.

Electronic Cigarette Refill Liquids: Nicotine Content, Presence of Child-Resistant Packaging, and in-Shop Compounding.

PURPOSE: To expand on our 2015 study of the nicotine content accuracy of e-liquids, including salts, and the presence of child-resistant packaging. We also describe compounding in shop (CIS). DESIGN AND METHODS: We analyzed samples from 35 shops. CIS processing was observed. Descriptive statistics summarized the data, and inference was performed. RESULTS: Actual nicotine content was significantly less than the identified content, on average, with a mean percent deviation 34.0% below the identified content. Only 3.8% of the samples' actual nicotine content was within 10% of the identified content; the maximum deviation was 213.2%. Of eight uniquely packaged samples, including designs resembling pop cans, ice cream cones, etc., the mean percent deviation was -39.6%; none were within 10% of the identified content. Eight shops compounded samples. After removing outlier values, significant differences were found in the percent deviations between the CIS and non-CIS free-base samples. A significantly higher percentage of CIS samples had nicotine content > 10% above the identified content, and none were within 10%. One shop visually estimated the nicotine quantities to add, e-liquids were not always relabeled to reflect new nicotine levels, and protective materials were not always worn during compounding. Child-resistant packaging was not present for one third of the samples. CONCLUSIONS: Labeling of nicotine content in e-liquids remains inaccurate, child-resistant packaging is inconsistent, and CIS is problematic. Effective e-liquid regulation is needed to protect public health. PRACTICE IMPLICATIONS: Nurses should educate families about the serious health risks of e-liquids and advocate for increased e-liquid regulations.

Compliance with North Dakota's smoke-free law among vape and tobacco specialty shops.

OBJECTIVE: To determine compliance with North Dakota's smoke-free law in vape shops and other tobacco specialty shops selling electronic nicotine delivery systems (ENDS) or e-liquids. DESIGN: In this 2019 descriptive study, shops (n = 35) were assessed for compliance with the smoke-free law by observation of indoor and outdoor areas for smoking or vaping, or evidence of such activity in prohibited areas, and the presence of required no-smoking signs. RESULTS: Only two shops (5.7%) were fully compliant with the smoke-free law. Full compliance for indoor and outdoor environments was 8.5% and 42.8%, respectively. Vaping occurred inside five shops (14.3%), and smoking occurred outdoors within required smoke-free areas in two (5.7%) shops. Four (11.4%) and 17 (48.6%) shops complied with indoor and outdoor signage requirements, respectively. CONCLUSIONS: Overall compliance remained low, although much of the noncompliance was related to signage. Use or evidence of ENDS use occurred both indoors and outdoors where prohibited by law. Classifying ENDS as tobacco products would require tobacco licensure of shops selling ENDS and e-liquids, aiding in identification of the shops for education and enforcement efforts to ensure compliance with the law and to improve public health protection.

Modifying SAMseq to account for asymmetry in the distribution of effect sizes when identifying differentially expressed genes.

RNA-Seq is a developing technology for generating gene expression data by directly sequencing mRNA molecules in a sample. RNA-Seq data consist of counts of reads recorded to a particular gene that are often used to identify differentially expressed (DE) genes. A common statistical method used to analyze RNA-Seq data is Significance Analysis of Microarray with emphasis on RNA-Seq data (SAMseq). SAMseq is a nonparametric method that uses a resampling technique to account for differences in sequencing depths when identifying DE genes. We propose a modification of this method that takes into account asymmetry in the distribution of the effect sizes by taking into account the sign of the test statistics. Through simulation studies, we showthat the proposed method, comparedwith the traditional SAMseqmethod and other existing methods provides better power for identifying truly DE genes or more sufficiently controls FDR in most settings where asymmetry is present. We illustrate the use of the proposed method by analyzing an RNA-Seq data set containing C57BL/6J (B6) and DBA/2J (D2) mouse strains samples.

Differences in Child Passenger Safety Counseling Frequency and Attitudes by Health Care Provider Specialty.

Many children are not being properly restrained in motor vehicles, resulting in unnecessary injury and fatalities. Health care provider (HCP) education is effective at increasing proper child restraint within vehicles. However, differences exist by HCP specialty in regards to frequency of child passenger safety (CPS) counseling. This study of a sample of 255 HCPs examined differences in CPS counseling by HCP specialty (pediatric vs. non-pediatric). HCPs from several upper Midwest states were surveyed about how frequently they provide CPS counseling in their practice by patient age and their attitudes toward CPS-related issues. Pediatric HCPs were twice as likely as non-pediatric HCPs to always provide CPS counseling to parents/guardians of children aged 5 or older. Non-pediatric HCPs were more likely than pediatric HCPs to feel that counseling is ineffective at increasing child seat/booster (p = 0.001) or seat belt use (p = 0.006). Non-pediatric HCPs were more likely than pediatric HCPs to feel there is inadequate time to provide CPS counseling in their practice setting (p = 0.001), and were less likely to know where to refer patients if they have questions regarding CPS issues (0.0291). The differences in HCP attitudes toward CPS counseling provision and the resulting differences in counseling frequency by patient age may contribute to disparities for patients who have limited or no access to pediatric HCPs. Additional research is needed to investigate the rationale for counseling differences seen by HCP specialty and patient age, and the potential effect on child motor vehicle injuries and fatalities.

An improved method for computing q-values when the distribution of effect sizes is asymmetric.

MOTIVATION: Asymmetry is frequently observed in the empirical distribution of test statistics that results from the analysis of gene expression experiments. This asymmetry indicates an asymmetry in the distribution of effect sizes. A common method for identifying differentially expressed (DE) genes in a gene expression experiment while controlling false discovery rate (FDR) is Storey's q-value method. This method ranks genes based solely on the P-values from each gene in the experiment. RESULTS: We propose a method that alters and improves upon the q-value method by taking the sign of the test statistics, in addition to the P-values, into account. Through two simulation studies (one involving independent normal data and one involving microarray data), we show that the proposed method, when compared with the traditional q-value method, generally provides a better ranking for genes as well as a higher number of truly DE genes declared to be DE, while still adequately controlling FDR. We illustrate the proposed method by analyzing two microarray datasets, one from an experiment of thale cress seedlings and the other from an experiment of maize leaves. AVAILABILITY AND IMPLEMENTATION: The R code and data files for the proposed method and examples are available at Bioinformatics online.

Development of a Flavor Ingredient Wheel Linking E-Liquid Additives to the Labeled Flavor of Vaping Products.

E-liquids contain combinations of chemicals, with many enhancing the sensory attractiveness of the product. Studies are needed to understand and characterize e-liquid ingredients, particularly flavorings, to inform future research and regulations of these products. We identified common flavor ingredients in a convenience sample of commercial e-liquids using gas chromatography-mass spectrometry. E-liquid flavors were categorized by flavor descriptors provided on the product packaging. A Flavor Ingredient Wheel was developed to link e-liquid flavor ingredients with flavor categories. An analysis of 109 samples identified 48 flavor ingredients. Consistency between the labeled flavor and ingredients used to produce such flavor was found. Our novel Flavor Ingredient Wheel organizes e-liquids by flavor and ingredients, enabling efficient analysis of the link between ingredients and their flavor profiles and allowing for quick assessment of an e-liquid ingredient's flavor profile. Investigating ingredient profiles and identifying and classifying commonly used chemicals in e-liquids may assist with future studies and improve the ability to regulate these products.

Bracketology in Pharmacy Education: The Impact of March Medication Madness on Student Engagement and Knowledge.

Description of the Problem. Gamification is used in pharmacy education as an innovative learning strategy to engage learners with educational content. The March Medication Madness activity used bracketology, a type of gamification not previously described in pharmacy education literature, to increase student engagement and knowledge of key disease states. The Innovation. The activity was developed for use in a capstone course during the final semester of the didactic pharmacy curriculum. Students created medication-related pearls that were placed in a tournament-style bracket. Students then completed brackets to predict the winning pearls and voted biweekly to determine the most clinically significant pearl. Student knowledge was assessed pre- and post-activity along with a post-activity perception assessment. Critical Analysis. Of the 52 student participant responses, most agreed or strongly agreed that the activity increased understanding and stimulated interest in course material, while adding a fun element to the course. There was a statistically significant increase (P = .002) in the average percentage of multiple-choice questions students answered correctly from the pre-test (57.7% ± 1.5%) to the posttest (63.1% ± 1.9%). Pearls that received the most votes were no more likely to be associated with an increase in knowledge than pearls receiving fewer votes. Next Steps. Implementation of a bracketology activity was perceived by students as fun, engaging, and beneficial in understanding course material. However, increase in knowledge was limited. This shows the importance of structuring gamification in a way that provides educational value and underscores the need to modify the activity to promote student learning.

Recombinant inbred line panels inform the genetic architecture and interactions of adaptive traits in Drosophila melanogaster.

The distribution of allelic effects on traits, along with their gene-by-gene and gene-by-environment interactions, contributes to the phenotypes available for selection and the trajectories of adaptive variants. Nonetheless, uncertainty persists regarding the effect sizes underlying adaptations and the importance of genetic interactions. Herein, we aimed to investigate the genetic architecture and the epistatic and environmental interactions involving loci that contribute to multiple adaptive traits using two new panels of Drosophila melanogaster recombinant inbred lines (RILs). To better fit our data, we re-implemented functions from R/qtl (Broman et al. 2003) using additive genetic models. We found 14 quantitative trait loci (QTL) underlying melanism, wing size, song pattern, and ethanol resistance. By combining our mapping results with population genetic statistics, we identified potential new genes related to these traits. None of the detected QTLs showed clear evidence of epistasis, and our power analysis indicated that we should have seen at least one significant interaction if sign epistasis or strong positive epistasis played a pervasive role in trait evolution. In contrast, we did find roles for gene-by-environment interactions involving pigmentation traits. Overall, our data suggest that the genetic architecture of adaptive traits often involves alleles of detectable effect, that strong epistasis does not always play a role in adaptation, and that environmental interactions can modulate the effect size of adaptive alleles.

An intravenous pancreatic cancer therapeutic: Characterization of CRISPR/Cas9n-modified Clostridium novyi-Non Toxic.

Clostridium novyi has demonstrated selective efficacy against solid tumors largely due to the microenvironment contained within dense tumor cores. The core of a solid tumor is typically hypoxic, acidic, and necrotic-impeding the penetration of current therapeutics. C. novyi is attracted to the tumor microenvironment and once there, can both lyse and proliferate while simultaneously re-activating the suppressed immune system. C. novyi systemic toxicity is easily mitigated by knocking out the phage DNA plasmid encoded alpha toxin resulting in C. novyi-NT; but, after intravenous injection spores are quickly cleared by phagocytosis before accomplishing significant tumor localization. C. novyi-NT could be designed to accomplish intravenous delivery with the potential to target all solid tumors and their metastases in a single dose. This study characterizes CRISPR/Cas9 modified C. novyi-NT to insert the gene for RGD, a tumor targeting peptide, expressed within the promoter region of a spore coat protein. Expression of the RGD peptide on the outer spore coat of C. novyi-NT indicates an increased capacity for tumor localization of C. novyi upon intravenous introduction based on the natural binding of RGD with the αvß3 integrin commonly overexpressed on the epithelial tissue surrounding a tumor, and lead to immune stimulation.

Neutralizing and protective murine monoclonal antibodies to the hemagglutinin of influenza H5 clades 2.3.2.1 and 2.3.4.4.

BACKGROUND: Highly pathogenic avian H5 influenza viruses have spread and diversified genetically and antigenically into multiple clades and subclades. Most isolates of currently circulating H5 viruses are in clade 2.3.2.1 or 2.3.4.4. METHODS: Panels of murine monoclonal antibodies (mAbs) were generated to the influenza hemagglutinin (HA) of H5 viruses from the clade 2.3.2.1 H5N1 vaccine virus A/duck/Bangladesh/19097/2013 and the clade 2.3.4.4 H5N8 vaccine virus A/gyrfalcon/Washington/41088-6/2014. Antibodies were selected and characterized for binding, neutralization, epitope recognition, cross-reactivity with other H5 viruses, and the ability to provide protection in passive transfer experiments. RESULTS: All mAbs bound homologous HA in an ELISA format; mAbs 5C2 and 6H6 were broadly binding for other H5 HAs. Potently neutralizing mAbs were identified in each panel, and all neutralizing mAbs provided protection in passive transfer experiments in mice challenged with a homologous clade influenza virus. Cross-reacting mAb 5C2 neutralized a wide variety of clade 2.3.2.1 viruses, as well as H5 viruses from other clades, and also provided protection against heterologous H5 clade influenza virus challenge. Epitope analysis indicated that the majority of mAbs recognized epitopes in the globular head of the HA. The mAb 5C2 appeared to recognize an epitope below the globular head but above the stalk region of HA. CONCLUSIONS: The results suggested that these H5 mAbs would be useful for virus and vaccine characterization. The results confirmed the functional cross-reactivity of mAb 5C2, which appears to bind a novel epitope, and suggest the therapeutic potential for H5 infections in humans with further development.

Strong concordance between transcriptomic patterns of spleen and peripheral blood leukocytes in response to avian pathogenic Escherichia coli infection.

Avian pathogenic Escherichia coli (APEC) causes morbidity in chickens and exhibits zoonotic potential. Understanding host transcriptional responses to infection aids the understanding of protective mechanisms and serves to inform future colibacillosis control strategies. Transcriptomes of spleen and peripheral blood leukocytes (PBLs) of the same individual birds in response to APEC infection were compared to identify common response patterns and connecting pathways. More than 100 genes in three contrasts examining pathology and infection status were significantly differentially expressed in both tissues and similarly regulated. Tissue-specific differences in catalytic activity, however, appear between birds with mild and severe pathology responses. Early expression differences, between birds with severe pathology and uninfected controls, in the mitogen-activated protein kinase pathway in PBLs precede spleen responses in the p53 and cytokine-cytokine receptor pathways. Tissue bianalysis is useful in identifying genes and pathways important to the response to APEC, whose role might otherwise be underestimated in importance.

Zebrafish embryos hatch early in response to chemical and mechanical indicators of predation risk, resulting in underdeveloped swimming ability of hatchling larvae.

Plasticity in hatching time allows embryos to maximize fitness by balancing the benefits and costs of remaining bound within the chorion against the benefits and costs of emerging as a free-swimming larva. Here, in the first experiment, we exposed zebrafish (Danio rerio) embryos to either chemical cues from crushed embryos (simulating egg predation) or to blank water control. Embryos exposed to alarm cues hatched sooner, and had shorter body lengths and underdeveloped fins, relative to larvae from the water treatment. Burst swimming speed was significantly slower for larvae that hatched from the alarm cue treatment than for larvae from the water treatment. In a second 2×2 experiment, we exposed zebrafish embryos to either chemical alarm cues from conspecific embryos, mechanical disturbance (magnetic stir bar) to simulate a predator probing the substrate for developing embryos, both chemical and mechanical indicators of risk, or neither (control). We found similar effects in terms of earlier time to hatch at an earlier stage of development and poorer swimming performance of hatchling larvae. In the second experiment, these effects occurred in response to mechanical disturbance with or without the presence of chemical alarm cues. Alarm cues alone produced no effects in the second experiment. Taken together, these data indicate that zebrafish embryos demonstrate a facultative trade-off between risk of predation acting on two stages of their life history.

A Pilot Study of Blood-Based Methylation Markers Associated With Pancreatic Cancer.

Over the past several decades in the United States, incidence of pancreatic cancer (PCa) has increased, with the 5-year survival rate remaining extremely low at 10.8%. Typically, PCa is diagnosed at an advanced stage, with the consequence that there is more tumor heterogeneity and increased probability that more cells are resistant to treatments. Risk factors for PCa can serve as a way to select a high-risk population and develop biomarkers to improve early detection and treatment. We focus on blood-based methylation as an approach to identify a marker set that can be obtained in a minimally invasive way (through peripheral blood) and could be applied to a high-risk subpopulation [those with recent onset type 2 diabetes (DM)]. Blood samples were collected from 30 patients, 15 had been diagnosed with PCa and 15 had been diagnosed with recent onset DM. HumanMethylationEPIC Beadchip (Illumina, CA, United States) was used to quantify methylation of approximately 850,000 methylation sites across the genome and to analyze methylation markers associated with PCa or DM or both. Exploratory analysis conducted to propose importance of top CpG (5'-C-phosphate-G-3') methylation site associated genes and visualized using boxplots. A methylation-based age predictor was also investigated for ability to distinguish disease groups from controls. No methylation markers were observed to be significantly associated with PCa or new onset diabetes compared with control the respective control groups. In our exploratory analysis, one methylation marker, CpG04969764, found in the Laminin Subunit Alpha 5 (LAMA5) gene region was observed in both PCa and DM Top 100 methylation marker sets. Modification of LAMA5 methylation or LAMA5 gene function may be a way to distinguish those recent DM cases with and without PCa, however, additional studies with larger sample sizes and different study types (e.g., cohort) will be needed to test this hypothesis.

Spleen transcriptome response to infection with avian pathogenic Escherichia coli in broiler chickens.

BACKGROUND: Avian pathogenic Escherichia coli (APEC) is detrimental to poultry health and its zoonotic potential is a food safety concern. Regulation of antimicrobials in food-production animals has put greater focus on enhancing host resistance to bacterial infections through genetics. To better define effective mechanism of host resistance, global gene expression in the spleen of chickens, harvested at two times post-infection (PI) with APEC, was measured using microarray technology, in a design that will enable investigation of effects of vaccination, challenge, and pathology level. RESULTS: There were 1,101 genes significantly differentially expressed between severely infected and non-infected groups on day 1 PI and 1,723 on day 5 PI. Very little difference was seen between mildly infected and non-infected groups on either time point. Between birds exhibiting mild and severe pathology, there were 2 significantly differentially expressed genes on day 1 PI and 799 on day 5 PI. Groups with greater pathology had more genes with increased expression than decreased expression levels. Several predominate immune pathways, Toll-like receptor, Jak-STAT, and cytokine signaling, were represented between challenged and non-challenged groups. Vaccination had, surprisingly, no detectible effect on gene expression, although it significantly protected the birds from observable gross lesions. Functional characterization of significantly expressed genes revealed unique gene ontology classifications during each time point, with many unique to a particular treatment or class contrast. CONCLUSIONS: More severe pathology caused by APEC infection was associated with a high level of gene expression differences and increase in gene expression levels. Many of the significantly differentially expressed genes were unique to a particular treatment, pathology level or time point. The present study not only investigates the transcriptomic regulations of APEC infection, but also the degree of pathology associated with that infection. This study will allow for greater discovery into host mechanisms for disease resistance, providing targets for marker assisted selection and advanced drug development.

Too Much of a Good Thing? Cerebral Sinovenous Thrombosis Due to Excessive Milk Intake Associated Anemia.

INTRODUCTION: In young children, excessive cow's milk intake causes iron-deficiency anemia, which is associated with hypercoagulable states. We present a case series of 4 toddlers with excessive milk intake iron-deficiency anemia and cerebral sinovenous thrombosis. METHODS: Retrospective chart review of 4 patients was performed for patients with cerebral sinovenous thrombosis and iron-deficiency anemia secondary to excessive milk intake. Iron-deficiency anemia was defined as hemoglobin <11 mg/dL, mean corpuscular volume <70 fL, and serum ferritin <12 µg/L. Excessive milk intake was defined as consumption of >24 oz daily. Clinical, laboratory, and radiographic features were reviewed. RESULTS: Age ranged from 12 to 24 months. Average hemoglobin, hematocrit, mean corpuscular volume, and ferritin levels were 6.1 g/dL, 22.7 g/dL, 52.7 fL, and 3.2 ng/mL, respectively. Daily milk consumption ranged from 40 to 60 oz. All patients presented with focal neurologic deficits, including seizures in 3. The location of cerebral sinovenous thrombosis varied, and 3 patients had venous infarcts, one of them hemorrhagic. All patients had a limited diet and were described as "picky eaters" by their parents, and only 1 had transitioned of a bottle. All patients were treated with anticoagulation, iron supplementation, and extensive dietary counseling to reduce cow's milk intake. CONCLUSION: Iron-deficiency anemia due to excessive milk intake is an important and preventable etiology of pediatric cerebral sinovenous thrombosis. Focused anticipatory guidance is necessary for at-risk groups to prevent this neurologic emergency.

Glutathione S-Transferase pi-1 Knockdown Reduces Pancreatic Ductal Adenocarcinoma Growth by Activating Oxidative Stress Response Pathways.

Glutathione S-transferase pi-1 (GSTP1) plays an important role in regulating oxidative stress by conjugating glutathione to electrophiles. GSTP1 is overexpressed in breast, colon, lung, and prostate tumors, where it contributes to tumor progression and drug resistance; however, the role of GSTP1 in pancreatic ductal adenocarcinoma (PDAC) is not well understood. Using shRNA, we knocked down GSTP1 expression in three different PDAC cell lines and determined the effect on cell proliferation, cell cycle progression, and reactive oxygen species (ROS) levels. Our results show GSTP1 knockdown reduces PDAC cell growth, prolongs the G0/G1 phase, and elevates ROS in PDAC cells. Furthermore, GSTP1 knockdown results in the increased phosphorylation of c-Jun N-terminal kinase (JNK) and c-Jun and the decreased phosphorylation of extracellular signal-regulated kinase (ERK), p65, the reduced expression of specificity protein 1 (Sp1), and the increased expression of apoptosis-promoting genes. The addition of the antioxidant glutathione restored cell viability and returned protein expression levels to those found in control cells. Collectively, these data support the working hypothesis that the loss of GSTP1 elevates oxidative stress, which alters mitogen-activated protein (MAP) kinases and NF-κB signaling, and induces apoptosis. In support of these in vitro data, nude mice bearing orthotopically implanted GSTP1-knockdown PDAC cells showed an impressive reduction in the size and weight of tumors compared to the controls. Additionally, we observed reduced levels of Ki-67 and increased expression of cleaved caspase-3 in GSTP1-knockdown tumors, suggesting GSTP1 knockdown impedes proliferation and upregulates apoptosis in PDAC cells. Together, these results indicate that GSTP1 plays a significant role in PDAC cell growth and provides support for the pursuit of GSTP1 inhibitors as therapeutic agents for PDAC.