H2Oslim®, the water-soluble polysaccharides from Agaricus bisporus, significantly reduces body weight and improves lipid parameters in overweight humans in a randomized double-blind vs placebo study

In vitro tests demonstrated that the water-soluble polysaccharides from Agaricus bisporus is capable of trapping fat through a cationic polymer trapping net in contact with water. The fat trapping capability of this cationic net could favorably impact on blood lipid levels and diminishing cardiovascular risk factors. The aim of the study was to understand the effects of H2Oslim®, on body weight and associated metabolic parameters in overweight human volunteers. The study participants comprised of 100 healthy, overweight volunteers (defined as BMI 25-30 kg/m2) randomly divided into two groups. The groups received on a daily basis, either 400 mg of H2Oslim® or matching placebo in a double blinded. Participants were evaluated at baseline, 4, 8 and 12 weeks of the study. Significant improvements in body weight, waist circumference as well as plasma total cholesterol, LDL cholesterol, were observed in the H2Oslim® group (H-group) compared with the placebo group (P-group). The results here reported revealed that the supplementation of H2Oslim® seems a useful tool in dealing with obesity and hyperlipidemia


Introduction
Obesity is one of the major health concerns worldwide affecting approximately all physiological roles of the body. It increases the risk for multiple chronic conditions, such as cardiovascular disease [1,2], diabetes mellitus, different kinds of cancers [3], some musculoskeletal disorders [4], and poor mental health [5].
Also, studies show that obesity can have negative influences on the quality of life, healthcare costs and work productivity [6,7].
The World Health Organization (WHO) has estimated that obesity affects 500 million people worldwide and it could potentially increase to one billion people globally by 2030 [8,9]. The global obesity epidemic and ageing population are major public health concerns. With excess weight becoming increasingly 'normal', public perceptions of what constitutes a healthy body weight have become more inaccurate over time, with increasing numbers perceiving a body mass index (BMI) in the overweight or obese range (≥25 kg/m2) to be 'about right'. The numbers of older adults with overweight (BMI 25-29.9 kg/m2) and obesity (BMI ≥30 kg/ m2) are rising rapidly, due to concurrent increases in the number of adults who reach older age and the proportion who carry excess weight. Understanding how older adults perceive their own weight status, and the extent to which this is influenced by their age and health status, is important for informing targeted recommendations and interventions to promote healthy weight in later life. Although overweight and obesity result from a combination of causes, overconsumption of high-energy foods is considered as the primary cause of obesity [10].
In addition, evidence showed that factors such as lack of physical activity, lack of sleep, sedentary lifestyle and high level of the risk of obesity conjointly or independently. For example, an obesogenic behavior like lack of physical activity may be influenced by individual and social factors such as genetic, biological, marital, educational and occupational factors [15,16].The use of animalderived ingredients as chitosan and may other botanical extracts to diminish the lipid absorption has been widely reported [17][18][19][20][21][22][23][24][25][26].
The reduction of the adipose tissue seems a clear target to reduce obesity and obesity related diseases. The search of molecules capable to reduce the metabolic changes in plasma lipids, liver triglycerides leading to weight loss is a widely accepted approach to reduce the burden of obesity. Recently, it has been shown in vitro that, the water-soluble polysaccharides from common mushroom exerts a fat trapping activity on blood lipids through the formation of a cationic polymer capable to trap anionic complex substances as fat [27]. For such purpose, a double-blind vs placebo study was conducted to assess the safety and effectiveness of H2Oslim® on reducing cholesterols levels and weight in overweight healthy population.

Study design/intervention
The study was a randomized, double-blind vs placebocontrolled design for 12 weeks. Volunteers (44 men/ 56 women) were randomly divided into two groups -placebo (P-group) or H2OSLim (H-group). The participants consumed one capsule of placebo or one capsule of H2OSLim (containing 400 mg) 5-15 minutes before lunch throughout the study period.

Test materials
H2Oslim®, in high density form, was kindly provided by (mean BMI 28,6) were divided into 2 groups. 50 were randomly assigned to P-Group (placebo) and 50 to H-Group (H2Oslim®).
Of these initial 100 volunteers, 94 completed the 12-week study: 44 from the P-group, and 50 from the H group. Four volunteers dropped out of the study because they did not show a rapid weight loss (n = 4). There was not a reason given by the other two (n = 2).

Food Intake
The daily energy intake in both groups during the study was 2200-2700 kcal. Seven-day dietary and activity assessment of the subjects at baseline showed similar food intake habits and energy levels. No major dietary intervention or formal physical activity program was instituted during the course of the study; participants were instructed to maintain the current level of physical activity prior to study randomization during the course of the study

Sample collection
Fasting blood samples (5 ml of blood) were collected at baseline, and at 4, 8, and 10 weeks. Serum obtained from each blood sample was split into multiple 500 μl aliquots and stored at -20°C until needed for the measurement of total cholesterol and LDL cholesterol.

Primary outcome
Body Mass Index (BMI) and waist circumference was registered at the beginning, midpoint, and end of each intervention period by qualified staff. Systolic and diastolic blood pressures was also determined three times at 5-min intervals while volunteers are seated using a standardized mercury sphygmomanometer, to assess the ability of changes in blood pressure

Statistical analysis
Data analyses were collected at the end of each intervention; therefore, a replacement or substitution of a withdrawal subject was not planned; therefore, the missing data was considered unavailable data. The normality of variables as blood pressure were assessed. Variables not following a normal distribution will be transformed. Normal variables will be expressed as the mean and standard error of the mean. An intention-to-treat analysis will be performed for the analysis of efficacy. In order to determine differences in basal characteristics, a paired t-test will be used.
Carryover effects will be assessed by three-way ANOVA. A general lineal model for repeated measurements will be performed to evaluate differences between variables at the beginning, mid-point, and end of each intervention period. Two-way ANOVA was used to determine the influence of treatment (active or placebo) and time on the continuous dependent variables. Statistical analyses was performed using IBM SPSS statistics v [22]. The body weight and waist circumferences of the two groups shown no significant differences at baseline (Table 1). Baseline levels of serum lipids were similar in the two groups (Table 1).

Adverse events
The supplementation of H2Oslim® was well-tolerated.
Intestinal flatulence was reported in five participants, 2 in the P-Group (n=2) and 3 in the H-Group (n=3).

Results
The 100  in the H-group.

Discussion
The present study revealed that water soluble polysaccharides from Agaricus bisporus, Is a safe strategy in reducing body weight in overweight and/or obese subjects. The current clinical study shows that H2Oslim ® administration is associated with a decrease in the blood lipid levels in comparison with the placebo group.
These observations support the idea that the weight reduction effect of H2Oslim® is mediated by its direct effect on lipid trapping [27].Chitosan, the non-water soluble linear polysaccharide Although early studies conducted with hypocaloric diets suggested that chitosan could a significant effect on body weight, later and larger clinical trials (n=250) showed no effect at all when compared to a placebo in obese population [29]. Since then, the efficacy of chitosan has been investigated at different doses and also when administered concomitantly with a calorie-restricted diet and physical activity in various short-term and mediumterm studies, and therefore the effectivity of non-soluble linear polysaccharides has been disputed. By contrast, the results here presented show specifically that the water-soluble polysaccharides isolated from the common mushroom, Agaricus bisporus, do have a direct impact on weight loss and waist circumference.
The 12 weeks double blind vs placebo study revealed that a solely 400 mg dosage is capable of reducing significantly both variables with a deeper impact on waist circumference. This data together with the total cholesterol and ldl-cholesterol data analysis seems to reveal that the mechanism previously suggested about the fat trapping properties of the water-soluble polysaccharides net [27] is linked to the weight loss effect. According to previous studies [30], adipose tissue plays a prominent role in the clinical expression of metabolic syndrome, most likely mediated by the increased release and peripheral tissue action of non-esterified fatty acids and by the dysregulated production of adipocytesecreted proteins, including leptin, adiponectin, resistin, TNF-α, and IL-6. The observation of a significant weight loss, as well as a decrease in cholesterol levels is consistent with early reports in the literature observing the association [31]. The current results suggest that H2Oslim® may be a helpful adjunct in the management of overweight. Further studies need to be processed in order to understand the direct impact of the H2Oslim® intake together with a healthy lifestyle on overweight people. Additional studies are needed to corroborate these initial first results.