Impact of JuicePlus+ on the Health Status of an Overweight Stressed Population

An anticipated eighty critical care, female health care professionals will be recruited. This population has been shown to have a high occupational stress load and have increased risk for post-traumatic stress disorder (PTSD). The subjects will be premenopausal, overweight/obese (Body Mass Index -BMI : 25 - 40 kg/m2) and between the ages of 25-50 years.

Exclusion criteria include smoking of any sort, smokeless tobacco use, chronic or excessive alcohol consumption (>1 drink per day), pregnancy or lactation, recent surgery or illness, diabetes, heart disease, use of antibiotics or medications (other than medication deemed not to impact outcome measures in this protocol-medication use will be reviewed on a case by case basis by the principal investigator).

Health history, medication and dietary supplement questionnaires will be completed by all subjects to determine eligibility. A health and perceived stress questionnaire will also be completed at each sampling period. Characteristics of stool will also be recorded for the three days prior to sampling using the Bristol criteria.

All food and drink intake will be recorded for the 7-day period prior to providing the first sample (baseline). Subjects will repeat this procedure at each sampling period. During the initial recording period, subjects will be asked to consume foods that represent their usual diet, with no more than a combined 4 fruit and vegetable servings each day, and be instructed to attempt to mimic the same diet during the 7 day periods each sampling period.

Diet records will be analyzed using nutrition software and the data will be used for analysis purposes as needed. Subjects will be encouraged to consume their usual diet throughout the study period.

Subjects will be weighed and total body composition determined using Dual-energy X-ray absorptiometry (DXA) for baseline subject characterization. Prior to the DXA scan a pregnancy test will be performed to confirm that the subject is not pregnant.

Subjects will be randomly assigned to supplement or the placebo capsules. The intervention will start between days 1 and 5 of the menstrual cycle to avoid confounding factors due to menstrual cycle hormone variation. The capsules are to be taken with a meal in the morning and in the evening with 3 capsules at each time, for a total of 6 capsules/day for a 20 week period. At the beginning of the 17th week, subjects consuming the supplement will be randomized with one group adding the "Complete" meal replacement component that will replace the first meal of the day and the second group following their normal diet.

Initial Laboratory Visit: screening visit During the initial visit to the laboratory, subjects will complete the informed consent form, health history, medication and dietary supplement usage and perceived stress questionnaires. Subjects' heart rate and blood pressure, height, weight, waist, and hip circumference will be measured. A pregnancy test will be performed to confirm that the subject is not pregnant. Body composition will be determined via DXA to determine body composition, as increased visceral fat is causative in obesity-induced inflammation. Upon completion of this screening visit, subjects will be scheduled for their initial testing visit.

Test Visit Procedures For each condition (supplement and placebo), subjects will report to the lab a total of three times. Subjects that continue on with the "Complete" meal-replacement phase of the study will report to the lab one additional time. Subjects will collect their stool during the 24 hours prior to reporting to the lab on each test occasion (or as close as possible to that time point). Subjects will report to the lab in a 10 hour fasted state, without having performed strenuous activity within the prior 48 hours. Subjects will have been instructed to obtain at least 7 hours of sleep during the night prior to testing. No supplement or placebo will be taken prior to blood collection, as described below.

Stool Collection Stool samples will be collected by subjects during the 24 hours immediately before reporting to the lab on each test occasion (or as close as possible to this time point). Subjects will be instructed to line the pail with the biohazard bag, complete a bowel movement in the bag, seal the bag with the zip tie, and place the bag in the insulated cooler bag with two freezer packs. The pail, biohazard bag, zip tie, insulated cooler and freezer packs will be provided by the investigator. Verbal and written instructions for collection, storage, and delivery of the stool samples will be available to each participant. Upon receipt by investigators, these samples will be stored at -70 degrees Celsius until processing. All sample handling will occur in the fume hood and universal precautions will be observed to minimize the risk of infection. Following publication of the study results, remaining stool samples will be disposed of properly as biohazard waste.

Blood Collection Blood samples (approximately 20 ml; 4 teaspoons) will be taken from subjects via venipuncture (by someone skilled in collecting blood). After the first blood draw, subjects will consume a cocktail of glucose, lactulose and mannitol. Another blood sample (3 ml) will be collected 90 minutes after ingestion of this cocktail. The purpose of consuming this cocktail and collecting a second blood sample is to determine the impact of the treatment on glucose tolerance and also its effect on intestinal permeability.

The collected blood samples will be processed and stored in multiple aliquots at -70 degrees Celsius until analyzed. Following publication of the study results, remaining blood samples will be disposed of properly as biohazard waste. Universal precautions will be observed to minimize the risk of infection or injury to either the subject or the phlebotomist. Potential risks for this procedure include soreness around the sampling site, bruising, and infection.

White Blood Cell Harvest White blood cells will be harvested from the collected whole blood using a density gradient (Ficoll) and stored in multiple aliquots at -70 degrees Celsius until analyzed. Following publication of the study results, remaining cell samples will be disposed of properly as biohazard waste. Universal precautions will be observed to minimize the risk of infection or contamination.

Dietary Intake and Records All subjects will be asked to consume their usual diet throughout the study period and be instructed to attempt to mimic the same diet during the 7 days prior to sampling. Subjects will record all food and drink consumed during the 7 days prior to each sampling day.