Proteomics analysis reveals a potential new target protein for the lipid-lowering effect of Berberine8998.

Berberine8998 is a newly synthesized berberine derivative with better lipid-lowering activity and improved absorption. The objective of this study was to investigate the effects of berberine8998 on serum cholesterol and lipid levels in vivo and to examine the mechanisms involved. Hamsters on high-fat diet (HFD) were administered berberine or berberine8998 (50 mg·kg-1·d-1, ig) for 3 weeks. Berberine8998 administration significantly lowered the total cholesterol, triglycerides and LDL-C levels in HFD hamsters. Bioinformatics revealed that berberine and berberine8998 shared similar metabolic pathways and fatty acid metabolism was the predominant pathway. Western blot validation results showed that peroxisomal acyl-coenzyme A oxidase 1 (ACOX1) and long-chain fatty acid-CoA ligase 1 (ACSL1), two proteins involved in fatty acid metabolism, were expressed differently in the berberine8998 group than in the untreated group and the berberine treatment group. Biochemistry results showed that berberine8998 significantly lowered the non-esterified fatty acid (NEFA) levels, which may lead to a reduction in TG levels in the berberine8998 treatment group and the differences observed in proteomics analyses. Pharmacokinetic analysis conducted in rats. After administration of berberine or berberine8998 (50 mg/kg, ig), berberine8998 exhibited a remarkably improved absorption with increasing bioavailability by 6.7 times compared with berberine. These findings suggest that berberine8998 lowers cholesterol and lipid levels via different mechanisms than berberine, and its improved absorption makes it a promising therapeutic candidate for the treatment of hypercholesterolemia and obesity.

Cell-free DNA as a molecular tool for monitoring disease progression and response to therapy in breast cancer patients.

Due to the spatial and temporal genomic heterogeneity of breast cancer, genomic sequencing obtained from a single biopsy may not capture the complete genomic profile of tumors. Thus, we propose that cell-free DNA (cfDNA) in plasma may be an alternate source of genomic information to provide comprehensive data throughout a patient's clinical course. We performed a retrospective chart review of 100 patients with stage 4 or high-risk stage 3 breast cancer. The degree of agreement between genomic alterations found in tumor DNA (tDNA) and cfDNA was determined by Cohen's Kappa. Clinical disease progression was compared to mutant allele frequency using a two-sided Fisher's exact test. The presence of mutations and mutant allele frequency was correlated with progression-free survival (PFS) using a Cox proportional hazards model and a log-rank test. The most commonly found genomic alterations were mutations in TP53 and PIK3CA, and amplification of EGFR and ERBB2. PIK3CA mutation and ERBB2 amplification demonstrated robust agreement between tDNA and cfDNA (Cohen's kappa = 0.64 and 0.77, respectively). TP53 mutation and EGFR amplification demonstrated poor agreement between tDNA and cfDNA (Cohen's kappa = 0.18 and 0.33, respectively). The directional changes of TP53 and PIK3CA mutant allele frequency were closely associated with response to therapy (p = 0.002). The presence of TP53 mutation (p = 0.0004) and PIK3CA mutant allele frequency [p = 0.01, HR 1.074 (95 % CI 1.018-1.134)] was excellent predictors of PFS. Identification of selected cancer-specific genomic alterations from cfDNA may be a noninvasive way to monitor disease progression, predict PFS, and offer targeted therapy.

Heparin-induced thrombocytopenia with very high antibody titer is associated with slower platelet recovery and higher risk of thrombosis.

Heparin-induced thrombocytopenia (HIT) is a prothrombotic disorder mediated by platelet-activating antibodies targeting platelet factor 4 (PF4) and heparin complex. A higher antibody titer is reflected in a higher optical density (OD) by enzyme-linked immunosorbent assay for heparin-PF4 antibodies. This single-institution retrospective study of 116 HIT patients examined the effect of heparin-PF4 OD on time to platelet recovery, vascular thrombosis, and in-hospital mortality. Patients were divided into 3 cohorts based on heparin-PF4 OD: cohort 1 had an OD ≥ 2 and ≤ 2.4, cohort 2 had an OD > 2.4 and ≤ 2.8, and cohort 3 had an OD > 2.8. A higher OD titer was associated with significantly increased time to platelet recovery when compared between cohorts 1 versus 2 (HR = 0.599, p = 0.0221) and 1 versus 3 (HR = 0.515, p = 0.0014), as well as an increased risk of thrombosis (79.4%-cohort 3 vs 53.8%-cohort 2 vs 46.1%-cohort 1, p = 0.04), but had no impact on mortality (2.62-alive vs 2.65-deceased, p = 0.7432). A higher OD titer can inform risk assessment and support decision-making in HIT patients; however, prospective studies are needed to further clarify the impact of heparin-PF4 OD on outcomes.

Perceptions of physically active men with prostate cancer on the role of physical activity in maintaining their quality of life: possible influence of androgen deprivation therapy.

OBJECTIVE: The primary aim of this study was to examine the perceptions of older men with prostate cancer regarding their quality of life and physical activity post-diagnosis, and the potential benefits and risks associated with being physically active. A secondary aim was to gain some preliminary insight into how these perceptions may differ as a function of androgen deprivation therapy (ADT). METHODS: Two focus groups were conducted, consisting of six ADT and eight non-ADT men, respectively. The probe questions used assessed the link between quality of life and physical activity as well as the benefits and risks associated with physical activity. Data were transcribed verbatim and themes identified using a general inductive thematic approach. RESULTS: The primary themes identified were sexual health, 'plumbing' and non-urogenital side-effects, return to and increased levels of physical activity post-diagnosis, physical health/function and psychological benefits of physical activity as well as over-doing it and age-related risks of excessive physical activity. However, not all themes were present in both the ADT and the non-ADT sub-groups. CONCLUSIONS: These results further highlight the link between physical activity and quality of life in prostate cancer survivors and how they use physical activity as a part of their survivorship process. Of particular interest was how several men on ADT used resistance training to counteract ADT-related side-effects affecting their masculinity. As the evidence for physical activity for prostate cancer survivorship is increasing, cancer clinicians and service providers should consider ways to better assist these men, especially those on ADT become more active.

The long-term effects of a primary care physical activity intervention on mental health in low-active, community-dwelling older adults.

OBJECTIVES: To examine the effect that physical activity delivered via two different versions of the Green Prescription (a primary care physical activity scripting program) had on depressive symptomatology and general mental health functioning over a 12-month period in non-depressed, low-active, community-dwelling older adults. METHOD: Two hundred and twenty-five participants from the Healthy Steps study took part in the present study. Healthy Steps participants were randomized to receive either the standard time-based or a modified pedometer-based Green Prescription. Depression, mental health functioning and physical activity were measured at baseline, post-intervention (3 months post-baseline) and at the 9-month follow-up period. RESULTS: At post-intervention, a positive association was found between increases in leisure-time physical activity and total walking physical activity and a decrease in depressive symptomatology (within the non-depressed range of the GDS-15) and an increase in perceived mental health functioning, regardless of intervention allocation. These improvements were also evident at the follow-up period for participants in both intervention allocation groups. CONCLUSION: Our findings suggest that the standard time-based Green Prescription and a modified pedometer-based Green Prescription are both effective in maintaining and improving mental health in non-depressed, previously low-active older adults.

Perceived barriers, benefits, and motives for physical activity: two primary-care physical activity prescription programs.

This study examined whether perceived barriers, benefits, and motives for physical activity differed based on allocation to 2 different types of primary-care activity-prescription programs (pedometer-based vs. time-based Green Prescription). Eighty participants from the Healthy Steps study completed a questionnaire that assessed their perceived barriers, benefits, and motives for physical activity. Factor analysis was carried out to identify common themes of barriers, benefits, and motives for physical activity. Factor scores were then used to explore between-groups differences for perceived barriers, benefits, and motives based on group allocation and demographic variables. No significant differences were found in factor scores based on allocation. Demographic variables relating to the existence of chronic health conditions, weight status, and older age were found to significantly influence perceived barriers, benefits, and motives for physical activity. Findings suggest that the addition of a pedometer to the standard Green Prescription does not appear to increase perceived motives or benefits or decrease perceived barriers for physical activity in low-active older adults.

Creation and validation of a ligation-independent cloning (LIC) retroviral vector for stable gene transduction in mammalian cells.

BACKGROUND: Cloning vectors capable of retroviral transduction have enabled stable gene overexpression in numerous mitotic cell lines. However, the relatively small number of feasible restriction enzyme sequences in their cloning sites can hinder successful generation of overexpression constructs if these sequences are also present in the target cDNA insert. RESULTS: Utilizing ligation-independent cloning (LIC) technology, we have modified the highly efficient retroviral transduction vector, pBABE, to eliminate reliance on restriction enzymes for cloning. Instead, the modified plasmid, pBLIC, utilizes random 12/13-base overhangs generated by T4 DNA polymerase 3' exonuclease activity. PCR-based introduction of the complementary sequence into any cDNA of interest enables universal cloning into pBLIC. Here we describe creation of the pBLIC plasmid, and demonstrate successful cloning and protein overexpression from three different cDNAs, Bax, catalase, and p53 through transduction into the human prostate cancer cell line, LNCaP or the human lung cancer line, H358. CONCLUSIONS: Our results show that pBLIC vector retains the high transduction efficiency of the original pBABE while eliminating the requirement for checking individual cDNA inserts for internal restriction sites. Thus it comprises an effective retroviral cloning system for laboratory-scale stable gene overexpression or for high-throughput applications such as creation of retroviral cDNA libraries. To our knowledge, pBLIC is the first LIC vector for retroviral transduction-mediated stable gene expression in mammalian cells.

Healthy Steps trial: pedometer-based advice and physical activity for low-active older adults.

PURPOSE: We compared the effectiveness of 2 physical activity prescriptions delivered in primary care--the standard time-based Green Prescription and a pedometer step-based Green Prescription--on physical activity, body mass index (BMI), blood pressure, and quality of life in low-active older adults. METHODS: We undertook a randomized controlled trial involving 330 low-active older adults (aged =65 years) recruited through their primary care physicians' patient databases. Participants were randomized to either the pedometer step-based Green Prescription group (n = 165) or the standard Green Prescription group (n = 165). Both groups had a visit with the primary care practitioner and 3 telephone counseling sessions over 12 weeks aimed at increasing physical activity. Outcomes were the changes in physical activity (assessed with the Auckland Heart Study Physical Activity Questionnaire), blood pressure, BMI, quality of life (assessed with the 36-Item Short Form Health Survey), physical function status (assessed with the Short Physical Performance Battery), and falls over a 12-month period. RESULTS: Of the patients invited to participate, 57% responded. At 12 months, leisure walking increased by 49.6 min/wk for the pedometer Green Prescription compared with 28.1 min/wk for the standard Green Prescription (P=.03). For both groups, there were significant increases across all physical activity domains at 3 months (end of intervention) that were largely maintained after 12 months of follow-up. BMI did not change in either group. Significant improvements in blood pressure were observed for both groups without any differences between them. CONCLUSIONS: Pedometer use resulted in a greater increase in leisure walking without any impact on overall activity level. All participants increased physical activity, and on average, their blood pressure decreased over 12 months, although the clinical relevance is unknown.

Cost-effectiveness of pedometer-based versus time-based Green Prescriptions: the Healthy Steps Study.

This paper reports on the cost-effectiveness of pedometer-based versus time-based Green Prescriptions in improving physical activity and health-related quality of life (EQ-5D) in a randomised controlled trial of 330 low-active, community-based adults aged 65 years and over. Costs, measured in $NZ (NZ$1=A$0.83, December 2008), comprised public and private health care costs plus exercise-related personal expenditure. Based on intention-to-treat data at 12-month follow up, the pedometer group showed a greater increase in weekly leisure walking (50.6 versus 28.1min for the time-based group, adjusted means, P=0.03). There were no significant between-group differences in costs. The incremental cost-effectiveness ratios, for the pedometer-based versus time-based Green Prescription, per 30min of weekly leisure walking and per quality-adjusted life year were, (i) when including only community care costs, $115 and $3105, (ii) when including only exercise and community care costs, $130 and $3500, and (iii) for all costs, -$185 and -$4999, respectively. The cost-effectiveness acceptability curves showed that the pedometer-based compared with the time-based Green Prescription was statistically cost-effective, for the above cost categories, at the following quality-adjusted life year thresholds: (i) $30000; (ii) $30500; and (iii) $16500. The additional program cost of converting one sedentary adult to an active state over a 12-month period was $667. The outcomes suggest the pedometer-based Green Prescription may be cost-effective in increasing physical activity and health-related quality of life over 12 months in previously low-active older adults.

General practitioners' views and experiences of counselling for physical activity through the New Zealand Green Prescription program.

BACKGROUND: Regular physical activity is beneficial in both the prevention and management of chronic health conditions. A large proportion of adult New Zealanders, however, are insufficiently active. To help increase population levels of physical activity in New Zealand the Green Prescription, a primary care physical activity scripting program, was developed. The primary aim of this study was to identify why general practitioners (GPs) counsel for physical activity and administer Green Prescriptions. A secondary aim was to examine GPs' views and experiences of Green Prescription counselling for the management of depression. METHODS: Individual face-to-face interviews were conducted with 15 GPs. All interviews were audio-taped and transcribed. Data were analysed using an inductive thematic approach. RESULTS: Several themes and sub-themes emerged from the data. Notably, GPs counselled for physical activity and prescribed Green Prescriptions for both primary preventive (e.g., weight control) and secondary management (e.g., diabetes management) purposes. GPs reported the benefits of the Green Prescription centred around two main themes: (i) a non-medication approach to a healthier lifestyle and (ii) the support benefits of physical activity. Time constraints within the consultation was the only main theme that emerged regarding the barriers GPs perceived to Green Prescription use. Physical activity in general, and physical activity prescribed through the Green Prescription, were also viewed by GPs as beneficial for the management of depression. CONCLUSIONS: The results of this study suggest that New Zealand GPs view the Green Prescription program as beneficial for their patients with pre-existing conditions and/or weight problems. While this is encouraging, the Green Prescription may also be used to promote physical activity in currently healthy but low-active and sedentary individuals. Such individuals are currently disease free, but are at risk for future health-related problems because of their inactive lifestyle. It is recommended that time constraints of the consultation in regard to administering Green Prescriptions could be dealt with by delegating the more time consuming tasks to the patient support counsellors that support the Green Prescription program, and having practice nurses assist in the administration of Green Prescriptions. Green Prescription counselling in conjunction with antidepressant medication may be beneficial for the management of depression and warrants further research.

Stable molluscicide formulation of an aqueous extract of Euphorbia myrsinites.

Aqueous extracts of Euphorbia myrsinites L. (Euphorbiaceae) were tested for molluscicidal activity against Biomphalaria glabrata; LC50 values of 15.1 and 8.9 ppm were obtained for the stem and leaf extracts, respectively, which are within the WHO limit for an effective molluscicide. However, the extracts were found to be unstable at room temperature and the level of activity fell rapidly (about 50% after 7 days at 20 °C). Moreover, the extracts were shown to be cytotoxic and would thus, if used as a molluscicide, be potentially hazardous to the user. As a result, attempts were made to produce a simple formulation that was stable and easy to handle, thereby reducing the danger to the user, but which would decompose rapidly in aqueous solution after application, thus reducing any lasting ecological damage. A product based on spray drying the aqueous extract of the leaves and stems led to a preparation that, although slightly lower in activity than the original extract, was stable at room temperature for at least a year, but which decomposed in aqueous solution at a similar rate to the original extract.

Practicing in Both Worlds: Why two General Practitioners Decided to Become Traditional Chinese Medicine Practitioners.

In New Zealand, little is known about Western medicine practitioners, such as general practitioners (GPs), who have also trained in traditional Chinese medicine (TCM). The aim was to identify the reasons why two GPs decided to study TCM and become integrated health care practitioners. Data from two Auckland-based integrated (combined GP and TCM) health care practitioners were analyzed for the present study. Individual face-to-face interviews were conducted. An inductive thematic approach was used to analyze the data. One main theme emerged. Both participants discussed how they perceived certain limitations in relation to their practice within the Western medicine model and that TCM allowed them to provide a holistic form of treatment. Being an integrated health care practitioner was perceived by participants as allowing them to treat the whole person rather than just the symptoms of a condition. TCM was also perceived to be beneficial as it allowed participants to provide alternative methods in treating their patients.

Barriers to physical activity in prostate cancer survivors.

AIMS: Despite the benefits of regular physical activity (PA), many prostate cancer (PCa) survivors are not engaging in sufficient PA to achieve health-related gain. This qualitative study sought to gain further insight regarding barriers to PA in older-aged PCa survivors. METHODS: Sixteen participants were individually interviewed, and data were analysed using an inductive thematic approach. RESULTS: Six main themes affecting perceived barriers for PA post diagnosis were identified: the effects of the PCa and PCa treatments on PA, urinary incontinence and bowel control, pre-existing comorbid conditions, increased age, time constraints and lack of proximity to PA or exercise venues. CONCLUSIONS: Only two of the six barriers identified directly related to having had PCa. With an increase in PCa survivorship, an active focus needs to be placed on the role that PA can have in helping maintain and improve both the physical and psychological health-related outcomes of PCa survivors.

Suppression of thioredoxin-1 induces premature senescence in normal human fibroblasts.

Thioredoxin (TRX) is a ubiquitous multifunctional thiol protein that is critically involved in maintaining cellular redox homeostasis. Levels of thioredoxin-1 (TRX1), the major isoform of TRX, have been shown to correlate with organismal lifespan and age-associated tissue deterioration. Accordingly, we investigated the direct functional effects of suppressing TRX1 levels on cellular senescence, a phenomenon intimately linked with tissue degeneration and aging. Here we find that suppression of TRX1 expression via shRNA rapidly induces premature senescence in young human skin fibroblasts through upregulation of the p53/p21(Cip1/Waf1) and p16(INK4a) tumor suppressor pathways. Moreover, inhibition of these pathways by introduction of SV40 Large T Antigen prevents TRX1 suppression-induced premature senescence but not susceptibility to oxidative stressors. Thus our results suggest that TRX1 has a role in suppressing senescence in normal cells in addition to its function as a redox-protective protein.

The Integrated Treatment for Chronic Pain and Anxiety in a Patient with Breast Implant Illness Symptoms: A Case Report.

INTRODUCTION: Women with breast implants may experience breast implant illness (BII). BII is comprised of a collection of systemic symptoms, such as fatigue and pain, that are postulated to be related to having breast implants. CASE PRESENTATION: This case report documents the naturopathic treatment of a 35-year-old female patient diagnosed with fibromyalgia, tension headaches, and chronic anxiety, perceived to be linked to and exacerbated by her textured cohesive gel implants. The patient sought naturopathic treatment for support during the explant process of her breast implants.Naturopathic treatment was provided over a 3-month period in conjunction with prescribed medication for her anxiety and tension headaches. CONCLUSION: At 3 months post-naturopathic treatment, the patient experienced a reduction in anxiety, fatigue, and self-reported pain. The integration of alternative therapies with prescribed pharmaceutical treatment appeared to positively impact the health of this patient.

Older adults' evaluations of the standard and modified pedometer-based Green Prescription.

INTRODUCTION The Green Prescription is a primary care programme designed to increase physical activity in individuals with low activity levels. Older adults tend to engage in insufficient physical activity to obtain health-related gain. AIM To examine participants' ratings of the Healthy Steps intervention and to assess how participants rated the use of a pedometer-based Green Prescription in aiding their physical activity. METHODS In total, 330 community-dwelling older adults who have low levels of activity were randomised to receive either a standard time-based Green Prescription or a modified pedometer-based Green Prescription. Post-intervention, 259 participants completed the participant evaluation questionnaire via postal survey. Data were analysed using descriptive statistics and Chi-squared analyses. RESULTS The standard components of the Green Prescription (general practitioner consultations and telephone counselling) received similar and higher ratings across both allocation groups than the use of print materials. A pedometer-based Green Prescription was rated as being helpful in aiding physical activity. DISCUSSION This study supports the importance of general practitioners' initial role in prescribing physical activity for older adults and of ongoing telephone support for longer-term adherence. Incorporating a pedometer can be effective in helping low-active older adults initiate and maintain regular physical activity.

The healthy steps study: a randomized controlled trial of a pedometer-based green prescription for older adults. Trial protocol.

BACKGROUND: Graded health benefits of physical activity have been demonstrated for the reduction of coronary heart disease, some cancers, and type-2 diabetes, and for injury reduction and improvements in mental health. Older adults are particularly at risk of physical inactivity, and would greatly benefit from successful targeted physical activity interventions. METHODS/DESIGN: The Healthy Steps study is a 12-month randomized controlled trial comparing the efficacy of a pedometer-based Green Prescription with the conventional time-based Green Prescription in increasing and maintaining physical activity levels in low-active adults over 65 years of age. The Green Prescription interventions involve a primary care physical activity prescription with 3 follow-up telephone counselling sessions delivered by trained physical activity counsellors over 3 months. Those in the pedometer group received a pedometer and counselling based around increasing steps that can be monitored on the pedometer, while those in the standard Green Prescription group received counselling using time-based goals. Baseline, 3 month (end of intervention), and 12 month measures were assessed in face-to-face home visits with outcomes measures being physical activity (Auckland Heart Study Physical Activity Questionnaire), quality of life (SF-36 and EQ-5D), depressive symptoms (Geriatric Depression Scale), blood pressure, weight status, functional status (gait speed, chair stands, and tandem balance test) and falls and adverse events (self-report). Utilisation of health services was assessed for the economic evaluation carried out alongside this trial. As well, a process evaluation of the interventions and an examination of barriers and motives for physical activity in the sample were conducted. The perceptions of primary care physicians in relation to delivering physical activity counselling were also assessed. DISCUSSION: The findings from the Healthy Steps trial are due in late 2009. If successful in improving physical activity in older adults, the pedometer-based Green Prescription could assist in reducing utilisation of health services and improve cardiovascular health and reduction of risk for a range of non-communicable lifestyles diseases. TRIAL REGISTRATION: Australian and New Zealand Clinical Trials Registry ACTRN012606000023550.

Patients' reasons for seeking traditional Chinese medicine: a qualitative study.

INTRODUCTION Acupuncture is one of the most well-known and utilised forms of traditional Chinese medicine (TCM). Very little is known about the factors that influence individuals to seek this type of treatment. The aim of this study was to identify and examine patients' reasons for seeking TCM treatment, which was mainly in the form of acupuncture. METHODS Participants were 15 Auckland-based patients who attended a student acupuncture clinic. Participants were individually interviewed. An inductive thematic approach was used to analyse data. RESULTS Three main themes were identified regarding factors that influenced individuals to seek TCM: the perceived limitations of pharmaceutical treatment for certain conditions; a previous positive experience of acupuncture; and word-of-mouth regarding the perceived effectiveness of acupuncture in the treatment of certain conditions. DISCUSSION Users perceive TCM-based acupuncture to be an effective form of treatment for their condition, especially after having previously received conventional medical treatment. Past and present users of TCM appear to have an advocate-based role with family and friends regarding the promotion of TCM-based acupuncture.

Influences on health-care practitioners' promotion of physical activity to their patients with prostate cancer: a qualitative study.

INTRODUCTION Physical activity is beneficial for the physical and psychological health of patients with prostate cancer (PCa). Health-care practitioners are ideally positioned to promote physical activity to their patients. AIM To identify factors that influenced health care practitioners to either promote or not promote physical activity to their patients with PCa. METHODS Individual interviews were conducted with 16 Auckland-based health-care practitioners, including specialists (oncologists and urologists), physiotherapists and complementary and alternative (acupuncturists) health-care practitioners. Data were analysed using an inductive thematic approach. RESULTS Treatment-related factors (ie counteracting side-effects of hormone suppression treatment), longer life expectancy and risk factors for other conditions appeared to influence the promotion of physical activity to patients. Time constraints of consultations and complex medical issues were barriers to the promotion of physical activity. CONCLUSIONS This study found that a variety of health-care practitioners are providing some degree of physical activity advice to their patients with PCa. Collaborative practice among health-care practitioners to verbally reinforce the benefits of physical activity, coupled with referral to experts in physical activity promotion/rehabilitation (such as physiotherapists), should be encouraged for best practice care.

Validation of RAPID E. coli 2 for enumeration and differentiation of Escherichia coli and other coliform bacteria in selected foods--Performance-Tested Method 050601.

RAPID'E. coli 2 agar (Bio-Rad Laboratories, Hercules, CA) is a chromogenic medium for differentiation and enumeration of E. coli and non-E. coli coliform bacteria in food. The principle of RAPID'E. coli 2 medium relies on simultaneous detection of 2 enzymatic activities, P-D-glucuronidase (GLUC) and beta-D-galactosidase (GAL). Coliforms, other than E. coli (GAL+/GLUC-), form blue to green colonies, whereas, specifically, E. coli (GAL+/GLUC+) form violet colonies. Eleven foods (raw ground beef, raw boneless pork, fermented sausage, processed ham, processed turkey, frozen turkey breast, raw ground chicken, cottage cheese, ricotta cheese, raw milk, and dry infant formula) were validated, comparing the performance of RAPID'E. coli 2 agar to the reference method AOAC 966.24. Two sample incubation temperatures were evaluated, 37 and 44 degrees C, testing a mixture of naturally and artificially contaminated foods. If naturally contaminated food was not available, the matrix was artificially inoculated with one E. coli strain and one non-E. coli coliform strain. Method comparison studies demonstrated some statistical differences between the 2 methods, which are expected when a plating method is compared to a most probable number method. Inclusivity and exclusivity rates of the medium were 99 and 94%, respectively. The RAPID'E. coli 2 method was shown to be stable when minor variations were introduced.