Additionally, all parameters of DNA damage significantly declined by nearly half of the initial value. The DNA damage frequency results together with significant apoptosis results are shown in Figure 1a (MNi), Figure 1b (NBs), Figure 1c (NPBs), and Figure 1d (apoptosis frequency). The BMR difference was a minimum of 28 and maximum of 303 kcal day−1, with an average of 90 kcal day−1. Calorie values measured and calculated with the bioelectrical impedance method reported in Table 2 served for the comparison of the values calculated from FETA software based on the EPIC questionnaires filled out by participants and reported in Table 3. G—gender; f—female; m—male; S—smokers; DMT2—Diabetes mellitus type 2; y—years; tbl—tablets; and CI—calorie daily intake (kcal day−1) from the EPIC food questionnaire calculated with FETA software. Since we recruited patients for this study in 2019 and 2020, after the diet they continued to be included in the program and observed. This study did not find significant correlations between urea levels before and after the VLCD with the level or decrease of permanent DNA damage. For a 2-week 600 kcal day−1 VLCD used as a pre-treatment before bariatric surgery, there was a BMI reduction of 1.60 kg m−2 (range—0.20 to 3.10), lean body mass loss of 2.80 kg, and fat mass loss of 1.70 kg, with the mean loss of 4.5 kg body weight (range—0.30 to 9.50) and 8.80% of excess body weight (range—0.90 to 17.10) after those 2 weeks . Our previous study (Ožvald et al. 2021, under revision) demonstrated the usefulness of the same diet on lowering the level of primary (repairable) DNA damage and primary oxidative DNA damage assessed with alkaline and Fpg alkaline comet assay . This is the first study on the assessment of the level of permanent genomic stability parameters in a 3-week hospital-controlled VLCD. People who lose weight quickly are also more likely to gain back the weight quickly. Losing more than 1 or 2 pounds (0.5 to 1 kg) a week is not safe for most people. You should only follow one of these diets with the help of your provider. 4. Biochemical Parameters The questionnaire was adjusted to Croatian food and portions, with more than 130 FFQ items included (see Supplementary Materials for the original FFQ questionnaire in English and the Croatian version used in the Croatian study; some food types were difficult to translate into English and were thus left in Croatian). The latter approval is mandatory for all experiments involving human exposure at the national level. As similar DNA damage and repair mechanisms are expected in different tissues 2,3, peripheral lymphocytes can serve as an excellent cell target due to their half-life and their presence in all body districts 19,20. The weight loss is a bigger stress for the body, and the hormonal response to the weight loss is much stronger. The types of rapid weight loss diets are described below. These diets are most often chosen by people with obesity who want to lose weight quickly. No difficulty was observed in maintaining weight for 8 weeks on 1500 kcal/day. An itemized receipt will be mailed to each participant weekly.Www.eatright.org/health/wellness/weight-and-body-positivity/4-ways-low-calorie-diets-can-sabotage-your-health.In most cases, an individual may choose New Direction initially and then transition into our second program–the Outlook program–when they are ready to begin utilizing food on a daily basis while losing weight.Some fad diets also severely limit calories to achieve rapid weight loss.The average initial weight of the whole cohort was 99.1 kg (SD 16.6).The study also demonstrated that participants who showed a decrease in GSH concentrations after the diet also had lower DNA damage, while this was not found in participants who did not have a reduction in GSH values.It is not the intent of this program to create an ideal BMI but to help reduce medical risk through weight reduction. This involves 2 days a week of fasting or VLCD and 5 days a week of eating your normal diet. These diets are often used before weight-loss surgery. A VLCD is only recommended for adults who have obesity and need to lose weight for health reasons. These diets are only to be used for a short time and are usually not recommended for more than several weeks. Rapid weight loss may not be safe for some people to do on their own. 6. Antioxidative GSH and Oxidative ROS Damage Measurement A participant’s weight is assessed by zones—Green Safety Zone, Yellow Caution Zone or Red Correction Zone—which then direct how frequently one should attend after the initial 12-week course. Physician review of blood glucose logs and adjustment of medications throughout the weight loss phase is included as medical monitoring. Exclusive use of meal replacement product to create a safer (compared to food-based) VLCD, mild ketosis, consistent weight loss results and avoidance of nutritional deficiencies. If you have a health condition, it is a good idea to talk with your provider before starting this or any diet plan to lose weight. Although a very-low-calorie diet (VLCD) is considered safe and has demonstrated benefits among other types of diets, data are scarce concerning its effects on improving health and weight loss in severely obese patients. Metabolic rate recovered to 93 per cent of prediet values by the end of 8 weeks of weight maintenance on 1500 kcal/day. Our diet program decreased BMR in all study participants, though in varying amounts, demonstrating highly individualized differences and no connections with other measured parameters in this study. The study also demonstrated that participants who showed a decrease in GSH concentrations after the diet also had lower DNA damage, while this was not found in participants who did not have a reduction in GSH values. An itemized receipt will be mailed to each participant weekly. All interested applicants are encouraged to attend an information meeting so that they are full informed about program options and fees. Most often goal weights fall into a BMI range of or, in some cases, a reduction of 20-30% from initial weight. On-going Support is offered with a free monthly support group meeting and a free weekly weigh-in clinic every Monday. With a growing body of studies on the efficacy and acceptability of VLCDs in overweight and obese patients, VLCDs have been considered and confirmed as effective and safe options for weight loss in the treatment of obese patients . Indeed, 15 study participants had lower GSH levels at the end of the diet, and in general they also had a lower apoptotic frequency (non-significant) and MN frequency (significant) (see Figure 2a,b) when compared to those who did have a reduction in GSH levels after the diet. Since there were different responses to GSH decreases after the diet and higher GSH values can be caused both genetically and by the type of the food eaten before the VLCD, particular attention was given to those participants who showed reduced serum GSH levels after the diet. Necrotic cell frequency slightly decreased, but most importantly, the frequency of cells entering apoptosis (programed cell death) significantly decreased by half after the diet, demonstrating the positive effect of VLCDs on genome stability. Statistically significant differences before and after the diet on the group level are marked in bold. That was the reason for not changing their regular therapy during the diet, and for those for whom it proved a necessity to change it, they were excluded from the study. Since the group included severely obese patients, it was expected that most, if not all, will have health problems, comorbidities, and diseases, etc. The EPIC-Norfolk food questionnaire was analyzed in this study only for daily calorie intake with FETA software to compare the values with the calculated basal metabolism rate before the start of the VLCD. The frequency of apoptotic and necrotic cells in 1000 lymphocytes per subject was also scored, and the scored DNA damage parameter frequency (per 2000 binucleated lymphocytes (BN)) was calculated and also expressed as 1000 BN. Only the study of Donmez-Altuntas et al. showed a significantly elevated frequency of apoptotic and necrotic cells as cytotoxicity markers in peripheral blood lymphocytes of obese and total overweight/obese subjects compared with normal-weight subjects, but that study did not use diet intervention . It has been demonstrated that if weight loss is induced by an increase in energy expenditure or metabolic rate, there may be an elevation in the mitochondrial production of ROS in the cells, thereby leading to higher ROS levels, which does not reflect the increase in DNA damage 60,61. Although our previous study with VLCDs using the comet assay and Fpg comet assay demonstrated a decrease in oxidative DNA damage levels, the increased serum ROS values after the VLCD this time were unexpected . In most cases, these diets are not sustainable for long enough to cause long-term weight loss. Some fad diets also severely limit calories to achieve rapid weight loss. The hormonal response is one of the reasons that weight loss slows down over time and also why weight gain occurs when the diet is stopped or relaxed. Following an average 2 kg weight regain within the first week of maintenance, there was no further weight regain. It has been argued that excessive lean body mass is lost with very restricted energy intake regimens which compromises metabolic rate and sabotages weight maintenance. The challenge to maintain lost weight is particularly relevant for advocates of VLCD, since these induce a high rate of weight loss. Rapid weight loss is more about cutting calories than exercising. For most people, it is safest to choose a diet in which you lose a 1/2 pound to 1 pound (225 grams to 500 grams) a week. Once you stop the diet, you are at risk for regaining the weight if you return to your old eating habits. Diets that incorporate fasting can cause rapid weight loss. Seventy-one per cent of the weight loss was fat. Both groups underwent a pedometer-based walking program; however only RT performed resistance training 3 days/week for 12 weeks. Total weight loss (TWL) encompassed pre- and postoperative WL. We analysed its impact on weight loss and other outcomes post-BS. Data Availability Statement These results will facilitate the further development of the program at the Duga Resa Special Hospital for Extended Treatment so that in the future it will also include 50 g of high-quality proteins and 55 g of carbohydrates per day, and adequate vitamin and mineral supplementation regimens during the VLCD. Further studies will be conducted on a higher number of volunteers, and will include other biomarkers, especially biomarkers of oxidative stress and antioxidants, with continued surveillance of subjects who have already entered the program. A recent review characterized DNA damage in obesity as an initiator, promoter, and predictor of cancer and advocated for early, pre-malignant assessment of genome integrity and stability to develop surveillance strategies and interventions . If you have a health condition, it is a good idea to talk with your provider before starting this or any diet plan to lose weight.The positive correlation of a decrease of GSH and decrease of DNA damage after our VLCD program confirms those findings.All interested applicants are encouraged to attend an information meeting so that they are full informed about program options and fees.The weight loss is a bigger stress for the body, and the hormonal response to the weight loss is much stronger.This is partly because some animal and human studies have shown benefits to fasting for people with diabetes and obesity.The New Direction© Program is a medically-supervised Very Low Calorie Diet (VLCD) which utilizes a meal replacement product exclusively to meet an individual’s nutritional needs; no food is consumed while on this program.Similarly, a non-significant decrease after our diet was observed for the leukocyte count (another inflammatory biomarker). This need was recognized by the Duga Resa Special Hospital for Extended Treatment, Croatia, which developed a local program that is also open to accepting obese persons from other parts of Croatia and other regions. Results of CBMN parameter frequency before (B) and after (A) the 3-week 567… Genomic instability parameters were lowered by half, reaching reference values usually found in the healthy population. Further, those clients who would prefer to attend for a longer period of time or at a frequency of their own choosing will be able to do so at a reduced cost. Adapting or Transition Phase is a series of four weeks of guided, structured meal plans which accomplishes a gradual transition from meal replacements to a full-food plan. A long-term weight management plan will include whole grains, fruits, vegetables, low-fat protein sources and regular exercise, in addition to the use of meal replacement products IF helpful specific to an individual’s needs. Most of the other patients’ medications were changed after the study at the individual level, based on the analysis of data created during the study. The impacts of the urea concentrations on multiple inner-organ DNA damage, and with oxidative DNA damage levels, have been demonstrated in other studies . As for the non-significant decrease in CRPs, Lips et al. demonstrated that a significant decrease in patient obesity can occur only after 3 months on a VLCD . We found a decrease in lipid parameters (TG, TC, HDL-C, and LDL-C), though with a maintained HDL-C/LDL-C ratio before and after the diet, similarly as reported elsewhere . This study aimed to evaluate permanent DNA damage during the VLCD using CBMN assay parameters and to assess possible correlations with changes in other anthropometric and biochemical parameters during the diet. The values for vitamin B12 and folic acid (measured to check for possible vitamin and mineral deficiency as an exclusion factor—results not shown) did not differ before and after the diet. The proliferation rate (NDI and M2 cells frequency) increased slightly but not significantly. Final statistics were taken or calculated from 1000 cells (Table 5) or 1000 BN cells (DNA damage frequency, Figure 1). Similar results in the decrease of glucose, insulin, and HOMA-IR levels in our VLCD were found in another study with obese patients and rapid weight loss with VLCDs . To the best of our knowledge, this is the first study involving human patients undergoing a 3-week 567 kcal hospital-controlled VLCD assessing genomic (in)stability through the CBMN assay together with anthropometric and biochemical parameters, and which compares these parameters before the start and on the last day of the diet period at both group and individual levels. As part of the personalized weight loss program developed at the Duga Resa Special Hospital for Extended Treatment, Croatia, we evaluated anthropometric, biochemical, and permanent DNA damage parameters (assessed with the cytochalasin B-blocked micronucleus cytome assay—CBMN) in severely obese patients (BMI ≥ 35 kg m−2) after 3-weeks on a 567 kcal, hospital-controlled VLCD. As part of the personalized weight loss program developed at the Duga Resa Special Hospital for Extended Treatment, Croatia, we evaluated anthropometric, biochemical, and permanent DNA damage parameters (assessed with the cytochalasin B-blocked micronucleus cytome assay-CBMN) in severely obese patients (BMI ≥ 35 kg m-2) after 3-weeks on a 567 kcal, hospital-controlled VLCD. Although the group was highly heterogeneous, consisting of patients with class II and III obesity, which can explain the lack of a significant difference in post-diet BMI (although there was a decrease), the results on DNA damage and selected biochemical parameters are relevant for the development of healthy weight loss strategies. This is the first study on the permanent genomic (in)stability in such VLCD patients. This process usually takes one to two weeks before a “start” appointment is set. If you decide New Direction is the right program for you, please call our office to request application forms to complete. On a VLCD, you may have as few as 800 calories a day and may lose up to 3 to 5 pounds (1.5 to 2 kg) week. People on these diets should be followed closely by a provider. These diets are less commonly recommended by health care providers. Very Low-Calorie Diet (VLCD) All participants except one outlier had a body fat mass decrease ranging from 2.20 to 8.30. The percentage loss of excessive body weight was highly individualized (one person lost almost 35%, one 27%, one 22%, three 20%, two 18%, four between 13 and 14%, two 11%, eight around 10%, one 8%, and one 6%). Individual values and differences in anthropometric measurements before and after the diet can be seen in Table 4. This remains controversial and contradicts current recommendations for slower weight loss. Emerging evidence suggests that rapid initial weight loss results in better long-term weight loss maintenance. Special attention will be also given to informing patients about the inflammatory potential of their habitual diet and the Dietary Inflammatory Index of specific foods, and to analyze the possible connections of these foods with the results . Thus, the CBMN assay can be used at both the group level and individual level, for an individualized or personalized medical approach in treating obesity. Fenech et al. in their review from 2020 demonstrated that the lymphocyte CBMN assay may prove to be a useful tool for the screening of obesity and the metabolic syndrome and its progression to diabetes and CVD in adults . We suggest all interested applicants attend a free monthly information session which offers a discussion and review of the New Direction program in greater detail. In most cases, if bariatric surgery is a covered benefit, then the physician’s appointments and labs on the VLCD program may be covered, too. Due to increasing awareness of obesity, though, some employers groups or insurers may begin to consider and include this type of benefit. Maximum weight loss was 14.40 kg, and the minimum weight loss was 3.80 kg, with an average loss of 9 kg at the group level. Other anthropometric factors did not significantly differ after the diet, although all demonstrated lower values (Table 3). The study revealed several possible biomarkers that could be used to track improvements of VLCDs, or even the patients that could be chosen for the program. VLCDs caused significant decreases in weight (loss), parameters of the lipid profile, urea, insulin resistance, and reduced glutathione (GSH). There is increasing evidence that CBMN parameters are linked to the pathogenesis of metabolic and cardiac diseases 15,16 and the severity of coronary artery disease 68,69. Considering that we analyzed the blood (lymphocytes) that circulated through different organs, the observed DNA damage with CBMN may reflect the overall condition of the body. Our future research will repeat these measurements and use more specific parameters (biomarkers) of oxidative damage and antioxidants to address these observed facts. In addition, this diet regime significantly decreased weight as an anthropometric parameter, blood glucose levels, HOMA-IR, TC, HDL-C, LDL-C, TG, and urea as biochemical parameters, and serum GSH levels, not only as an antioxidant but also as a parameter connected with higher metabolism rates, and almost significantly lowered BMI and insulin levels. Although the study recruited only 26 individuals, it revealed important results; a short 3-week 567 kcal VLCD in severely obese patients diminished the types of permanent DNA damage values by half. As oxidative stress response may affect DNA damage markers, insulin resistance and overall health status during body weight loss, serum ROS and GSH levels were determined and correlated with DNA damage parameters. The program includes personalized 24 h medical surveillance while severely obese patients are performing a 3-week 567 kcal hospital-controlled very-low-calorie diet (VLCD). In a highly heterogeneous group (class II and III in obesity, differences in weight, BMI, and other categories) consisting of 26 obese patients, the approach demonstrated its usefulness and benefits in health improvement, enabling an individual approach to further monitoring, diagnosis, treatment, and risk assessment based on changing anthropometric/biochemical VLCD parameters, and CBMN results. To lose weight this quickly you eat very few calories. VLCD did not produce losses of RMR beyond that expected from the loss of weight. It was observed that the metabolic rate dropped to 86 per cent of original by the end of the 8 weeks of VLCD. Body composition, resting energy expenditure (REE), neuromuscular function, and serum biomarkers were measured at weeks 0, 6, and 13. Preoperative WL compared weight before and after VLCD; postoperative WL compared post-VLCD weight and follow-up weight. Maintenance of weight loss with recovery of resting metabolic rate following 8 weeks of very low calorie dieting Finally, the most important findings of the study were the decreases in DNA damage values. The extent to which these molecular mechanisms of GSH and ROS contribute to obesity in humans and may be regulated through dietary means needs further research, though it has been demonstrated that elevated GSH reduces insulin sensitivity in adipocytes 62,63. Furthermore, VLCDs proved to be a useful approach in the weight treatment of obese patients with different comorbidities such as DMT2, and despite DMT2 being regarded as progressive and incurable, it appeared to be reversible by means of VLCD regimens, as claimed by other authors . Although obesity can lead to increased estrogen production, causing DNA damage via increased mitotic activity and/or directly via formation of DNA reactive metabolites, studies with similar restriction diets in rats demonstrated that sex hormones do not play a critical role in obesity-related DNA damage, oxidative base damage, or nucleotide excision repair (NER) 23,58. Although all patients in this study had similar conditions during the VLCD, the final results were highly individualized for each patient, with an individualized approach for each patient determining what they should do and accomplish further after the completion of the diet. We did not find any correlation with the change in other measured parameters in our study, and a similar trend was also observed in our previous study with VLCD and comet assay (Ožvald et al. 2021, under revision) . Increased fT4 but not fT3 levels suggests the presence of undefined factors stopping the conversion of elevated fT4 to fT3 levels in calorie restricted conditions or fasting . The slight fT4 increase and TSH and fT3 decreases observed here were also found in other short fasting diets . Both groups underwent a pedometer-based walking program; however only RT performed resistance training 3 days/week for 12 weeks.VLCD did not produce losses of RMR beyond that expected from the loss of weight.In addition, changes in medications were made for several participants at the end of the study due to an improvement in their health status, with dose reductions of antihypertensives and antidiabetics.Emerging evidence suggests that rapid initial weight loss results in better long-term weight loss maintenance.Although all patients in this study had similar conditions during the VLCD, the final results were highly individualized for each patient, with an individualized approach for each patient determining what they should do and accomplish further after the completion of the diet.The extent to which these molecular mechanisms of GSH and ROS contribute to obesity in humans and may be regulated through dietary means needs further research, though it has been demonstrated that elevated GSH reduces insulin sensitivity in adipocytes 62,63.Calorie values measured and calculated with the bioelectrical impedance method reported in Table 2 served for the comparison of the values calculated from FETA software based on the EPIC questionnaires filled out by participants and reported in Table 3. However, considering they walked usually once per day (if at all), both patients lowered the number of smoked cigarettes per day. Demographic information of the study participants including information on gender, age, medications, lifestyle factors, and calorie intake from the EPIC questionnaire are shown in Table 2. Spearman correlation was used for data correlation, with a significance level set at p ≤ 0.05. Anthropometric, biochemical, DNA damage, and oxidative damage parameters after descriptive statistics were analyzed using Mann–Whitney U-test. The cytokinesis-block proliferation index or nuclear division index (NDI) was calculated according to the following formula, with M1–M4 representing the number of cells with 1–4 nuclei, and N the total number of cells scored (1000) . After 72 h incubation, cells with the medium were transferred into glass centrifugation tubes, which were run at 450× g for 10 min in a centrifuge (Rotofix 32a, Hettich, Tuttlingen, Germany) using a swing bucket rotor. For each participant, the study started on the first day of hospitalization, when they signed the informed consent, filled out the questionnaires, anthropometric measurements were taken, and blood was drawn for biochemical and DNA damage analysis. Participants were recruited over several months due to limited hospital facilities, but all followed an identical diet regime. This study included severely obese patients with highly individualized ratios of different fat types and muscle loss, and in this short 3-week period nearly all of the assessed anthropometric parameters were observed to decline at the group level, although the difference was not statistically significant.A recent review characterized DNA damage in obesity as an initiator, promoter, and predictor of cancer and advocated for early, pre-malignant assessment of genome integrity and stability to develop surveillance strategies and interventions .As part of the personalized weight loss program developed at the Duga Resa Special Hospital for Extended Treatment, Croatia, we evaluated anthropometric, biochemical, and permanent DNA damage parameters (assessed with the cytochalasin B-blocked micronucleus cytome assay-CBMN) in severely obese patients (BMI ≥ 35 kg m-2) after 3-weeks on a 567 kcal, hospital-controlled VLCD.Our previous study (Ožvald et al. 2021, under revision) demonstrated the usefulness of the same diet on lowering the level of primary (repairable) DNA damage and primary oxidative DNA damage assessed with alkaline and Fpg alkaline comet assay .In the development of the most common NCDs and other chronic diseases and comorbidities, a key role is played by inflammation, oxidative and DNA damage accumulation in cells and organs, while several major NCDs (diabetes mellitus type 2 (DMT2), coronary heart disease, stroke, asthma, and several cancers) have been demonstrated to have an association with obesity and an unhealthy diet 2,3,4,5,6,7.You will not lose weight as fast with an LCD, but you can lose just as much weight with a VLCD.The DNA damage frequency results together with significant apoptosis results are shown in Figure 1a (MNi), Figure 1b (NBs), Figure 1c (NPBs), and Figure 1d (apoptosis frequency).Differences in MN, NB, NPB, and apoptotic frequency in different groups before the diet by age category.The study revealed several possible biomarkers that could be used to track improvements of VLCDs, or even the patients that could be chosen for the program. VLCDs also demonstrated other superior beneficial outcomes, such as more effective glycemic control, insulin resistance improvement, and improved lipid metabolism 25,26. Oxidative stress and inflammation, usually present in obesity, induce DNA damage, inhibit DNA damage repair, and cause disturbances in cell metabolism, promoting cancer growth (cancer cell proliferation and migration) and resistance to apoptosis . Pathophysiological mechanisms underlying obesity are explained with the increase of free fatty acids released from adipose tissue, lipid intermediates, insulin resistance with excess total and intra-abdominal adipose tissue, and inflammation 32,33,34,35,36,37. Micronucleus frequency can be affected by smoking status, gender, age, and vitamin concentration, especially vitamin B12 and folic acid. MN—micronucleus; MNi—micronuclei; BN—binucleated cell; MN1—BN with one MN; NBs—nuclear buds; 1NB—one NB; NPBs—nucleoplasmic bridges; NPB1—NB with one NPB; M1—mononuclear cells; M2—binuclear (BN) cells; M3—three-nuclear cells; M4—tetranuclear cells; and NDI—nuclear division index. This is partly because some animal and human studies have shown benefits to fasting for people with diabetes and obesity. For this diet, you have to eat all of your meals during an 8 hour period, for example, 10 am to 6 pm. This diet strategy is becoming more popular. You will not lose weight as fast with an LCD, but you can lose just as much weight with a VLCD. The S.T.A.R.® program (Steps To Avoid weight Regain) will be offered in 12-week sessions, resulting in a lower overall cost than the previous six-month (24-week) maintenance program. Average weight loss rate of 3-5 pounds weekly; women typically in the lower range and men in the upper range. If you are diabetic, please see the link below which describes the particular benefits of rapid weight loss, medication reductions and cost savings with a VLCD program. Www.eatright.org/health/wellness/weight-and-body-positivity/4-ways-low-calorie-diets-can-sabotage-your-health. In general, a rapid weight loss diet is not safe for children. In most cases, an individual may choose New Direction initially and then transition into our second program–the Outlook program–when they are ready to begin utilizing food on a daily basis while losing weight. In most cases, people will spend less on the VLCD program compared to normal weekly grocery and restaurant purchases! Program Costs will vary depending on individual calorie needs, the amount of weight to lose and the number of weeks on New Direction. It is not the intent of this program to create an ideal BMI but to help reduce medical risk through weight reduction. There was at least one patient who managed to regulate glycemic values after the diet, and the patient’s metformin therapy was cancelled.Similar results in the decrease of glucose, insulin, and HOMA-IR levels in our VLCD were found in another study with obese patients and rapid weight loss with VLCDs .Program Costs will vary depending on individual calorie needs, the amount of weight to lose and the number of weeks on New Direction.Average weight loss rate of 3-5 pounds weekly; women typically in the lower range and men in the upper range.None of these parameters demonstrated a correlation with BMI, BMR, HOMA-IR, or DNA damage parameters, before or after the diet.A total of 1000 lymphocytes per donor were scored to evaluate the frequency of cells with 1–4 nuclei (M1, M2, M3, and M4) within the same cytoplasm.After 72 h incubation, cells with the medium were transferred into glass centrifugation tubes, which were run at 450× g for 10 min in a centrifuge (Rotofix 32a, Hettich, Tuttlingen, Germany) using a swing bucket rotor.Patient consent was obtained from all subjects involved in the study and written informed consent has been obtained from the patient(s) to publish the results of this research. When study participants were divided into two equal-sized groups based on pre-diet GSH, the group with higher GSH levels had slightly lower DNA damage levels and higher apoptotic cell frequencies (Table 7). After the diet, the glucose levels were significantly lower and fell into the range of normal values. At the group level (Table 3), lipid profile parameters (TC, HDL-C, LDL-C, and TG) were all significantly lowered after the diet, with TC, LDL-C, and TG levels entering the normal interval reference range. Differences for other anthropometric parameters demonstrated once more that weight loss was highly individualized, with differing muscle and fat percentages, again highlighting the strong individual differences. In the development of the most common NCDs and other chronic diseases and comorbidities, a key role is played by inflammation, oxidative and DNA damage accumulation in cells and organs, while several major NCDs (diabetes mellitus type 2 (DMT2), coronary heart disease, stroke, asthma, and several cancers) have been demonstrated to have an association with obesity and an unhealthy diet 2,3,4,5,6,7. A total of 1000 lymphocytes per donor were scored to evaluate the frequency of cells with 1–4 nuclei (M1, M2, M3, and M4) within the same cytoplasm. An immunoassay analyzer ARCHITECT i1000sr (Abbott, Chicago, IL, USA) was used to measure insulin, TSH, fT3, and fT4 levels. Serum biochemical parameters at the beginning and end of the diet were selected as those expected to be possibly affected during this restricted diet and also connected with DNA (in)stability 23,24,25,26,27. The 3-week 567 kcal VLCD consisted of three daily meals freshly prepared by the hospital nutritionist, with special attention given to hydration conditions (mineral water was readily available during the hospital stay). The study excluded pregnant women, minors, legally incapacitated persons, patients with currently tumorous diseases, or in a diagnostic procedure with ionizing radiation. Possible Health Concerns Positive GSH change (decrease, contrary to a negative GSH change as an increase… The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results. A correlation was found between GSH decrease and reduced DNA damage. Similarly, a non-significant decrease after our diet was observed for the leukocyte count (another inflammatory biomarker). Our VLCD program did not cause any deaths or the need for any participants to withdraw from the study. Our results on anthropometric parameters are comparable, or preferably more beneficial than the results of other diets. However, due to rapid weight loss, VLCDs can cause complications including hypokalemia, cardiac arrhythmia, hyperuricemia, cholelithiasis, and mood and behavior alterations, and therefore should be performed only under strict and even 24 h medical surveillance 40,41. Considering CRP, HOMA-IR, or ROS values, the study group was too heterogeneous to draw any clear conclusions. An LCD is a better choice than a VLCD for most people who want to lose weight quickly. These diets usually allow about 1,000 to 1,200 calories a day for women and 1,200 to 1,600 calories a day for men. Most experts do not recommend using a VLCD for more than 12 weeks. The CBMN assay showed its usefulness as a genomic stability biomarker and demonstrated that a short 3-week 567 kcal VLCD can cut the types of permanent damage values by half. It seems that GSH is not only a cellular antioxidant that can decrease the level of oxidative stress and be protective against many diseases when present in elevated levels, but a decrease in GSH levels can increase energy expenditure, prevent obesity, and reduce insulin resistance . Due to NDI increase and changes/increases in cell counts after the VLCD (leukocytes, erythrocytes—results not shown in this study), it cannot be excluded that elevated ROS levels are not indicative of oxidative stress, but of higher metabolic activation in the cells connected with cellular repair, etc. We found that before the diet, 19 individuals had MN values above the median value of seven, while after the diet only four individuals had MN values above the median value. Differences in MN, NB, NPB, and apoptotic frequency in different groups before the diet by age category. Age did not show an effect on DNA damage parameters after the diet, however, differences in DNA damage were observed before the diet (Table 6). Importantly, the difference in DNA damage parameters before and after the diet was also confirmed with sex grouping. A word about meal replacements—while “tools” such as meal replacements, have been shown to be helpful in weight management, they are likely to be ineffective as a “single solution” without additional education and training. Contraindications for this program include liver or kidney disease, Type 1 diabetes, active ulcers or inflammatory bowel disease, bulimia, psychiatric disorders, pregnant or breastfeeding women and children or adolescents. Patients have access to our physicians by pager, if necessary, after hours and on weekends. Weekly medical monitoring is required and provided by our nursing staff who monitor blood pressure, weight changes, and potential side effects.which may include dizziness, fatigue, constipation, diarrhea, gallstones and hair loss. Preoperative Weight Loss via Very Low Caloric Diet (VLCD) and Its Effect on Outcomes After Bariatric Surgery Group BMI was significantly lower after the diet, while there was a mean decrease in body weight of 8 kg at the group level, or of 10 kg when medians were compared. At the group level, mean values for BMR and calculated calorie intake (CI) at the beginning of the study were similar, though the range of values demonstrated that several individuals did not report true portions and values of what they consumed in the dietary questionnaire, so the questionnaire details were not further analyzed in this study. Table 3 provides the group values of the anthropometric and biochemical parameters before and after the 3-week 567 kcal VLCD obtained from all 26 study participants. Necrotic cell frequency slightly decreased, but most importantly, the frequency of cells entering apoptosis (programed cell death) significantly decreased by half after the diet, demonstrating the positive effect of VLCDs on genome stability.Furthermore, VLCDs proved to be a useful approach in the weight treatment of obese patients with different comorbidities such as DMT2, and despite DMT2 being regarded as progressive and incurable, it appeared to be reversible by means of VLCD regimens, as claimed by other authors .Considering CRP, HOMA-IR, or ROS values, the study group was too heterogeneous to draw any clear conclusions.A decrease in DNA damage is usually followed by a decrease in ROS production, which was not the case in this study.The impacts of the urea concentrations on multiple inner-organ DNA damage, and with oxidative DNA damage levels, have been demonstrated in other studies .Obesity in Croatia is increasing rapidly, placing the country among those with the highest obesity prevalence in Europe 10,11,12,13.As part of the personalized weight loss program developed at the Duga Resa Special Hospital for Extended Treatment, Croatia, we evaluated anthropometric, biochemical, and permanent DNA damage parameters (assessed with the cytochalasin B-blocked micronucleus cytome assay—CBMN) in severely obese patients (BMI ≥ 35 kg m−2) after 3-weeks on a 567 kcal, hospital-controlled VLCD.The values for vitamin B12 and folic acid (measured to check for possible vitamin and mineral deficiency as an exclusion factor—results not shown) did not differ before and after the diet.If you are diabetic, please see the link below which describes the particular benefits of rapid weight loss, medication reductions and cost savings with a VLCD program. The New Direction© Program is a medically-supervised Very Low Calorie Diet (VLCD) which utilizes a meal replacement product exclusively to meet an individual’s nutritional needs; no food is consumed while on this program. Www.eatright.org/health/wellness/diet-trends/staying-away-from-fad-diets. 4 ways low-calorie diets can sabotage your health. This study aimed to evaluate the effects of the VLCD on markers of lipid profile, insulin action resistance, oxidative stress response by means of reactive oxygen species (ROS) and glutathione (GSH) levels, permanent DNA damage/stability, and anthropometric parameters in order to identify early biomarker(s) of patient status improvement during the 3-week VLCD. With the lack of serious systemic monitoring of overweight/obese people, there is a need to develop a special program for monitoring the health, diet, and lifestyle habits of the Croatian (obese) population that would also generate a personalized motivation towards weight loss lifestyle changes. Please check your Benefit Summary handbook for language referring to obesity treatment or weight loss programs; if such a benefit IS included, please provide notification at time of application. Rapid weight loss diet is usually for people who have health problems because of obesity. Rapid weight loss diet is a type of diet in which you lose more than 2 pounds (1 kilogram, kg) a week over several weeks. Epidemiological evidence reporting an increased incidence of all cancers for subjects with high CBMN parameters confirm that elevated levels of CBMN parameters play a causal role in cancer development 66,67. The value of this study is that other parameters such as NBs and NPBs, and the frequencies of apoptotic and necrotic cells, have not typically been measured in other studies. Changes in DNA damage levels (if any) were shown only after 6 months (slight) or after a year of diet or bariatric surgery (for details see reviews 14,15,23). It was observed that the metabolic rate dropped to 86 per cent of original by the end of the 8 weeks of VLCD.Differences in MN, NB, NPB, and apoptotic frequency in different groups before the diet by GSH level.Since the group included severely obese patients, it was expected that most, if not all, will have health problems, comorbidities, and diseases, etc.However, considering they walked usually once per day (if at all), both patients lowered the number of smoked cigarettes per day.The hormonal response is one of the reasons that weight loss slows down over time and also why weight gain occurs when the diet is stopped or relaxed.Serum biochemical parameters at the beginning and end of the diet were selected as those expected to be possibly affected during this restricted diet and also connected with DNA (in)stability 23,24,25,26,27.The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.Special attention will be also given to informing patients about the inflammatory potential of their habitual diet and the Dietary Inflammatory Index of specific foods, and to analyze the possible connections of these foods with the results . Even now, 2 years later, this patient continues to control DMT2 without any further medication, only with regular exercise, a prescribed diet, and regular check-ups. Weight loss results in a loss of metabolically active tissue, and therefore decreases BMR 50,51. In addition, changes in medications were made for several participants at the end of the study due to an improvement in their health status, with dose reductions of antihypertensives and antidiabetics. VLCDs are generally defined as a very low total energy intake of less than 800 kcal day−1 or 3.350 KJ day−1 for details see reviews 25, 26. Your provider may suggest waiting until you are on a more long-term diet to start exercising. Talk with your provider about what type of exercise you should do while you are on this type of diet. In some cases, these diets are not safe. The positive correlation of a decrease of GSH and decrease of DNA damage after our VLCD program confirms those findings. A decrease in DNA damage is usually followed by a decrease in ROS production, which was not the case in this study. Recent reviews demonstrated an improvement of subjective depressive symptoms following a VLCD program (for details see ). There was at least one patient who managed to regulate glycemic values after the diet, and the patient’s metformin therapy was cancelled. In a previous study on patients with DMT2, improved insulin sensitivity was the factor that improved glycemic control 56,57. This means that not only damaged cells with the loss of whole/part of the chromosomes were eliminated (decrease in MNi values and the number of BN cells with MNi), but also the number of cells with nuclear buds (amplification of excessive DNA) and nucleoplasmic bridges (cells with dicentric chromosomes) were diminished. The total values of the DNA damage parameters obtained by the CBMN assay are shown in Table 5 for 2000 binucleated cells (BN) per sample, including necrosis on 1000 counted cells and the results of the NDI for each category (M1–M4) on 1000 counted cells. HOMA-IR was also significantly lower after the diet, whereas insulin levels were on the borderline of significance. The maximum percentage of body weight loss was 9.60%, and the minimum was 3%, with an average of 6%. The average initial weight of the whole cohort was 99.1 kg (SD 16.6). Patient consent was obtained from all subjects involved in the study and written informed consent has been obtained from the patient(s) to publish the results of this research. The latter approval is mandatory at the national level for all the experiments involving human exposure. This research received no external funding except from the institutional funding of the Duga Resa Special Hospital for Extended Treatment, the Institute for Medical Research and Occupational Health, the University of Zagreb, and anonymous donations for the study. The CBMN assay measures changes in apoptotic, necrotic, and mitotic index frequency and shows different types of DNA damage through the frequencies of the micronuclei (MNi)- (chromosome breaks/loss during cell division), nucleoplasmic bridges (NPBs)- (chromosome rearrangement markers), and nuclear buds (NBs)- (DNA amplification markers) 17,18. All patients’ meals are prepared fresh in the hospital kitchen, supervised by the hospital nutritionist. Obesity in Croatia is increasing rapidly, placing the country among those with the highest obesity prevalence in Europe 10,11,12,13. In our previous study with the alkaline comet assay, obese subjects with the highest level of primary DNA damage had also the highest weight and BFM values before the diet and the highest change in the pre- and post-VLCD urea concentrations . This study included severely obese patients with highly individualized ratios of different fat types and muscle loss, and in this short 3-week period nearly all of the assessed anthropometric parameters were observed to decline at the group level, although the difference was not statistically significant. A longer VLCD also demonstrated a significant decrease in the BMI, VFL, and PBF parameters 48,49, which could also be projected for this VLCD program if the diet was to be applied for a longer time (more than 3 weeks). Regarding visceral abdominal fat decreases in obese subjects with DMT2, some VLCD studies have demonstrated a decrease after only 3 days together with body weight reduction 48,49, with further decreases of 25% or 55% after 6 or 16 weeks, respectively, along with a 16% or 45% decrease of abdominal subcutaneous fat, respectively 48,49. Mild restriction diets applied over longer periods (1750–2100 kcal daily, only a 500 kcal daily deficit more than the regular calorie intake in patients) resulted in a 3.5% loss of body weight (2 kg) in obese people, but only after 3 months, in addition to 10% after 6 months or 1 year, and a 17% body weight-loss only after 2 years of the diet 24,43. Differences in MN, NB, NPB, and apoptotic frequency in different groups before the diet by GSH level. We also compared obtained MN values before and after the diet with the established MN values in the healthy Croatian population (mean ± SD, median, range, and upper highest normal value 12.5 MN per 1000 BN cells). None of these parameters demonstrated a correlation with BMI, BMR, HOMA-IR, or DNA damage parameters, before or after the diet. TSH, fT3, and fT4 values were in the reference range both before and after the diet, though TSH and fT3 slightly decreased and fT4 slightly increased after the diet. There are studies demonstrating that lowering GSH values improves basal metabolism and weight loss.