1. INTRODUCTION

Herbal and dietary supplements are widely used. Weight loss supplements are popular and can be purchased easily from online shopping websites, pharmacies and local markets. Unfortunately, 80% of overweight individuals seeking weight loss depend on nonprescription herb products without the help of healthcare professionals (Blanck, et al.,2001).

These extensively marketed weight loss products promise a significant weight loss achievement, and consumers believe that herbal drugs and supplements are very safe and free from adverse effects, but researchers have found inconclusive evidence for the safety and effectiveness of many of the ingredients in such products (Johnston et al.,2016; Haaz et al., 2006 Saper et al.,2004; Allison et al.,2001;) . In addition many studies reported the potential toxicity of different herbal and plant extracts due to their harmful ingredients (Connolly et al., 1997; Eliason et al.,1994). The harmful ingredients can affect the vital organ tissue and induce toxicity to many organs such as the liver (Nigatu et al., 2017), kidney (Yang et al., 2018; Lai et al., 2009), heart (Eliason et al.,2012), spleen (Nigatu et al., 2017), and reproductive organs (Alraei, 2010). In addition, some weight loss supplements can lead to severe toxicity and even death (Roytman et al., 2014; Eliason et al.,2012).

A variety of over-the-counter weight loss supplements are widely available which include caffeine, chromium, ephedra, green tea, fucoxanthin guar gum, chitosan, conjugated linoleic acid, ginseng, glucomannan, hydroxy citric acid, L-carnitine, psyllium, and pyruvate (Saper et al.,2004; Lenz & Hamilton,2004).

Adios Grase, the herbal diet supplements are marketed as weight loss in Iraq, the UK, and some other countries (Relton et al.; 2014, Bello et al.; 2012 Rajashekar., 2009) for many years and are formally licensed as medicines with an authorized NHS numbers (LLRAPC, 2023; Bowdler, 2010; Rajashekar, 2009). Adios capsules contain the following ingredients: Fucus Dry Extract, Boldo Dry Extract, and Dandelion Root (Bowdler, 2010). Clinically, Fucus along with other chemicals is recommended as a nutritional treatment to lose weight among the overweight people (Vodouhè et al., 2022). In addition, Bolo and dandelion are widely used in folk medicine and are recognized as a medicinal herb in pharmacopoeia (Schütz,et al., 2006; Muñoz & Wilkomirsky ,2001).

Due to the lack of studies regarding the efficacy and side effects of these supplements, we conducted a study to explore the anti-obesity effects of adios supplements and also their effect on some of the biochemical, hematological and histological parameters of different organs: liver and kidney.

2. Materials and Methods

2.1 Animals of the study

Twenty-four healthy female albino rats (Rattus norvegicus) were used for the study. Animals were acquired from the animal house of the Biology Department, College of Science, University of Duhok. The animals were randomly divided into 4 groups: Control (T), low dose (LD), moderate dose (MD), and high dose (HD) groups. The animals were placed in ventilated cages made of polypropylene; five rats were kept in each cage under normal standard ambient conditions of temperature between 24-31 °C, humidity between 50%-55%, and a photoperiod of 12 h natural light and 12 h dark was maintained. The rats had access to a standard diet and water under ethical and standard laboratory conditions for 30 days (Maliakal & Wanwimolruk, 2001).

2.2 Weighting the rats

The weighing of rats was done using a weighing balance. Their weights ranged between 180-200 g. Weight recording was done every 5 days throughout the study.

2.3 Dosage, route, and duration of administration of Adios supplements.

Adios coated tablet (Aeropak-manufactured) was dissolved in distilled water and administrated to rats daily at three different concentrations (0.001g/100g), (0.002g/100g) and (0.003g/100g) as low (LD), medium (MD), and high (HD) concentrations, respectively. The control group was given only distilled water; each rat received 1ml of the dissolved concentration orally by gavage. Rats were weighed to adjust the dose administration for weight changes. The dose was administrated according to the dosages recommended by the manufacturer to the consumer (1 tablet 3 or 4 times a day). This experimental process lasted 30 days (Shin et al., 2021& Maeda et al., 2005).

2.4 Hematological, biochemical, and hormonal analyses.

On day 31, animals were anesthetized with diethyl ether. For hematological parameters, blood samples were obtained directly by heart puncture with 2 ml of blood collected in heparinized tubes and analyzed an automated hematological analyzer. The hematological parameters including the total count of red blood cells (RBC), white blood cell count (WBC), hemoglobin concentration (HGB), hematocrit (HCT), red cell distribution width (RDW), platelet count (PLT), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) were calculated. For biochemical and hormonal estimation, 5ml of blood was placed in a sterile gel tube and allowed to clot for 30 minutes before centrifugation at 4000 rpm for 15 minutes. Liver biochemical parameters included total protein, alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Kidney biochemical parameters included Albumin (ALB), Urea (URE), and Creatinine (Cr). All tests were performed by Cobas 600(C501), an automated chemistry analyzer, and measured according to their kit procedures depending on the absorbance photometry (enzymes, substrates, and specific proteins).

2.5 Histopathology Processing

After dissecting the animals, the liver and kidney were fixed in a 10% neutral buffered formalin solution. The samples were processed using routine histological processing method according to (Suvarna et al, 2018), thin sections (5 μM) were prepared for light microscopy studies and stained with hematoxylin (H) and eosin (E).

2.6 Statistical analysis

All statistical tests were performed with the statistical program SPSS 16.0. The results were expressed as mean ± standard errors. The one-way analysis of variance (ANOVA) was calculated using the software GenStat 18th (Lawes Agricultural Trust, VSN International Ltd, Oxford, UK). Tukey’s multiple range test. all groups were compared with the control group (pointed as *) (*P< 0.05), **= (P< 0.01), ***= (P< 0.001).

3. RESULTS

3.1 -The Effect of Adios Supplements on the Body Weight of Rats.

The body weight of each animal was recorded before the start of the adios administration; during the adios administration, animal weights were recorded every 5 days. Oral administration of adios with different concentrations and over 30 days showed no weight changes compared to the control group. Unexpectedly, a little weight gain was observed in the rats’ weights over time (within 1-15 days.) After 15 days, the weight of rats was stable in all the treated groups with no changes. Furthermore, all the treated groups (LD, MD, and HD) showed similar effects on rat body weight loss as shown in Figure (1).

DISCUSSION

Herb extracts and dietary supplements for weight loss or obesity management are attractive alternatives to the traditional therapy due to their accessibility to general publication. Many types of weight loss supplements are available in the markets and online stores, but, unfortunately, there is no or little knowledge about their efficacy, side effects, and toxicity is available. In Iraq and other countries, the use of adios herb extract supplements exists for more than tens of years and is very trendy among women wishing to lose weight. Unfortunately, based on our knowledge, no study has been conducted investigating the efficacy and effectiveness of adios supplements. Accordingly, testing those herb-based capsules and studying their effect was found to be necessary.

In the present study, the oral administration of (LD), (MD) and (HD) of the adios supplement has shown no efficacy in rat body weight loss during the 30 days of the experiment; this is inconsistent with some studies that used well-known fat-burn ingredients such as fucus extract, boldo, and dandelion for weight loss and obesity management (Vodouhè et al.,2023; Olas, 2022).

It is well documented that blood is an indicator of pathological and physiological status in humans and animals (Etim, et al., 2014). Furthermore, in acute toxicological studies, changes in hematological and biochemical parameters are usually used as indicators of toxicities. Adios showed significant decreases in WBC count and lymphocyte during the oral administration of (MD) and (HD) adios supplements. Additionally, our data suggested that adios supplement might cause iron deficiency as significant changes of low MCV and MCHC and high RDW with different concentrations observed. This data contrast with other studies which showed the antioxidant, antioxidant, and anti-inflammatory role of focus, boldo, and dandelion ( Olas, 2022; Shin et al., 2021; Catarino et al., 2018; Modaresi, & Resalatpour, 2012;). Our finding suggests that the adios herbs extract could have the immunosuppression side effect which causes leucopenia; this means that adios consumers could be more likely susceptible to infections and diseases with prolonged use.

Although the kidneys and liver have a cardinal role in metabolizing substances and excreting waste products like blood urea and creatinine, the serum biochemical measurement can also give an insight into the site of cellular tissue damage. However, adios showed some differences in their serum level (ALP, ALT, AST, urea, creatinine, and albumin), but it was not significant.

In this study, hematoxylin and eosin-stained sections of the kidney in the treated groups (LD, MD, and HD) show damage in the glomerulus, renal kidney, and blood vessels; this involved: increasing the periglomerular space, necrosis of glomeruli, hyperplasia of glomerulus, increasing the periglomerular space, hemorrhage among glomeruli, narrowing the lumen of some renal tubules, destruction of some renal tubules, and congestion of the blood vessels with markedly thick wall, and the presence of inflammation cells around the blood vessel. Nevertheless, in disagreement with our findings, Zhang and his colleagues showed a nephroprotective role of fucus in animal studies using models of kidney injury (Zhang et al., 2003; Miłek et al., 2019;). In addition, dandelion has nephroprotective features against kidney disorders (Karakuş et al., 2017; Lim et al., 2015).

Liver histology showed abnormal appearances such as degeneration of the hepatocytes, focal infiltration, branching, congestion, destruction of the central vein, presence of RBC, dilution of the sinusoid, vascular degeneration of hepatocytes, increase in the inflamed area of liver parenchyma, the presence of the fibroblast cells, dilation of the sinusoid, atrophy of hepatocyte cells pyknotic and dead hepatocytes, and steatosis with large lipid droplets are present in hepatocytes which displaced the nucleus to the cell periphery. Our findings disagree with those of Lim, Oles, Fernández, and their colleagues (Olas, 2022; Lim et al., 2015; Fernández et al., 2009), who described the protective changes induced by boldo, dandelion, and focus in liver pretreated with cytotoxic chemicals. However, our results can be supported by the number of reported cases of possible hepatotoxicity is increasing in patients consuming boldo as traditional medicine (Oliveria Sá, et al., 2020; Nortadas, & Barata, 2011 & Piscaglia et al., 2005).

The findings of the current study contradict most of those of the previous research studies. Focus, Dandelion, and Boldo were known to have many biological activities, such as anti-obesity (Vodouhè et al.,2022; Shin et al., 2021; Miłek et al., 2019; Maeda et al., 2005; Zhang et al., 2003) anti-inflammatory, anti-oxidant capacity, anti-diabetics, anti-tumor, chemoprotective and organ protective as well (Olas, 2022; Karakuş et al., 2017; Lim et al., 2015; Fitton et al., 2015; Ramos-Peralonso, 2014; Modaresi & Resalatpour, 2012).

This conflict in our results with other previous research could be due to the chemical composition used, all other studies have used only one component of adios herbs, with different concentrations.

Best of our knowledge, this is the first in vivo study on rats to investigate the physiological and histological effect of adios supplements.

The adverse effect of adios in this study could be a result of the absorption, metabolism, and accumulation of these chemicals in the body (Olas, 2022; Shin et al. 2021; Catarino et al., 2018; Karakuş et al., 2017, Ramos-Peralonso, 2014), or it might be a result of the incompatible herb-herb interaction (Che, 2013), since dandelion and boldo are recommended by scientists not to be mixed with other herbs (Armeller et al.,2014)

Moreover, some of the bioactive ingredients of dandelion and boldo contain toxic chemicals such as lectupirin, ascaridole, and boldine respectively, which could explain the adverse effect of adios in liver and kidney tissues (Gille et al., 2010; Basharat et al., 2009 & Leung, 1980).

Finally, these observations could suggest that adios supplements have low efficacy for weight loss and possibly have risk of toxic effects on the immune system of the experimental rats at different concentrations. Overweight individuals often try to lose weight are doing so without the help of the health care. Therefore, if healthcare professionals become more knowledgeable about the products available for treating obesity or overweight, then obese patients can be managed more safely and appropriately.

Conclusion:

Based on findings in this study, it can be concluded that adios supplements are not safe to use by women; they do cause immunosuppression and have zero efficacy in weight loss even with high concentrations.

Conflict of Interests: The authors declare that there are no competing interests associated with the manuscript.

Funding: This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

REFERENCES

Allison, D. B., Fontaine, K. R., Heshka, S., Mentore, J. L., & Heymsfield, S. B. (2001). Alternative treatments for weight loss: a critical review. Critical reviews in food science and nutrition, 41(1), 1–40. https://doi.org/10.1080/20014091091661.

Alraei, R. (2010). Herbal and Dietary Supplements for Weight Loss. Topics in Clinical Nutrition. 25. 136–150. 10.1097/TIN.0b013e3181dbb85e.

Armelle T. Mbaveng, Qiaoli Zhao, Victor Kuete. (2014). Harmful and Protective Effects of Phenolic Compounds from African Medicinal Plants.Toxicological Survey of African Medicinal Plants.Elsevier. Pages 577-609.ISBN 9780128000182.

Besharat S, Besharat M, Jabbari A Wild lettuce (Lactuca virosa) toxicity Case Reports (2009).bcr0620080134.

Bello, N., Winit-Watjana, W., Baqir, W., & McGarry, K. (2012). Disclosure and adverse effects of complementary and alternative medicine used by hospitalized patients in the North East of England. Pharmacy Practice, 10(3), 125–135.https://doi.org/10.4321/s1886-36552012000300002.

Blanck, H. M., Khan, L. K., & Serdula, M. K. (2001). Use of nonprescription weight loss products: results from a multistate survey. JAMA, 286(8), 930–935. https://doi.org/10.1001/jama.286.8.930.

Bonetti, G., Herbst, K. L., Donato, K., Dhuli, K., Kiani, A. K., Aquilanti, B., Velluti, V., Matera, G., Iaconelli, A., & Bertelli, M. (2022). Dietary supplements for obesity. Journal of preventive medicine and hygiene, 63(2 Suppl 3), E160–E168. https://doi.org/10.15167/2421-4248/jpmh2022.63.2S3.2757.

Catarino, M. D., Silva, A. M. S., & Cardoso, S. M. (2018). Phycochemical Constituents and Biological Activities of Fucus spp. Marine drugs, 16(8), 249. https://doi.org/10.3390/md16080249.

Che, C. T., Wang, Z. J., Chow, M. S., & Lam, C. W. (2013). Herb-herb combination for therapeutic enhancement and advancement: theory, practice and future perspectives. Molecules (Basel, Switzerland), 18(5), 5125–5141. https://doi.org/10.3390/molecules18055125.

Connolly, H. M., Crary, J. L., McGoon, M. D., Hensrud, D. D., Edwards, B. S., Edwards, W. D., & Schaff, H. V. (1997). Valvular heart disease is associated with fenfluramine-phentermine. The New England journal of medicine, 337(9), 581–588. https://doi.org/10.1056/NEJM199708283370901.

Eliason, B. C. , Doenier, A. J., Nuhlicek, D. N.(1994). Desiccated Thyroid in a Nutritional Supplement. J Fam Pract.(38) 3.

Eliason, M. J., Eichner, A., Cancio, A., Bestervelt, L., Adams, B. D., & Deuster, P. A. (2012). Case reports: Death of active-duty soldiers following ingestion of dietary supplements containing 1,3-dimethylamylamine (DMAA). Military medicine, 177(12), 1455–1459. https://doi.org/10.7205/milmed-d-12-00265.

Etim, N.N., Williams, M.E., Akpabio, U., & Offiong, E.E. (2014). Hematological Parameters and Factors Affecting Their Values. Agricultural Science, 2, 37-47.

Fernández, J., Lagos, P., Rivera, P. and Zamorano-Ponce, E. (2009), Effect of boldo (Peumus boldus Molina) infusion on lipoperoxidation induced by cisplatin in mice liver. Phytother.Res.,23:1024-1027. https://doi.org/10.1002/ptr.2746.

Fitton, J.H.; Stringer, D.N.; Karpiniec, S.S. (2015).Therapies from Fucoidan: An Update. Mar. Drugs 2015, 13, 5920-5946. https://doi.org/10.3390/md13095920.

Gille, L., Monzote, L., Stamberg, W., & Staniek, K. (2010). Toxicity of ascaridole from Chenopodium ambrosioides in mammalian mitochondria. BMC Pharmacology, 10(Suppl 1), A10. https://doi.org/10.1186/1471-2210-10-S1-A10.

Haaz, S., Fontaine, K. R., Cutter, G., Limdi, N., Perumean-Chaney, S., & Allison, D. B. (2006). Citrus aurantium and synephrine alkaloids in the treatment of overweight and obesity: an update. Obesity reviews: an official journal of the International Association for the Study of Obesity, 7(1), 79–88. https://doi.org/10.1111/j.1467-789X.2006.00195.x.

Karakuş, A., Değer, Y., & Yıldırım, S. (2017). Protective effect of Silybum marianum and Taraxacum officinale extracts against oxidative kidney injuries induced by carbon tetrachloride in rats. Renal failure, 39(1), 1–6. https://doi.org/10.1080/0886022X.2016.1244070.

Lai, M. N., Lai, J. N., Chen, P. C., Tseng, W. L., Chen, Y. Y., Hwang, J. S., & Wang, J. D. (2009). Increased risks of chronic kidney disease associated with prescribed Chinese herbal products suspected to contain aristolochic acid. Nephrology (Carlton, Vic.), 14(2), 227–234. https://doi.org/10.1111/j.1440-1797.2008.01061.x.

Leicester, Leicestershire and Rutland Area Prescribing Committee-NHS(n.d).Adios,Lastacess 12/12/2023.Aveilableon https://www.areaprescribingcommitteeleicesterleicestershirerutland.nhs.uk/search/?_search_terms=adios.

Lenz, T. L., & Hamilton, W. R. (2004). Supplemental products used for weight loss. Journal of the American Pharmacists Association: JAPhA, 44(1), 59–68. https://doi.org/10.1331/154434504322713246.

Lim, J.D.; Lee, S.R.; Kim, T.; Jang, S.-A.; Kang, S.C.; Koo, H.J.; Sohn, E.; Bak, J.P.; Namkoong, S.; Kim, H.K.; et al.(2015) Fucoidan from Fucus vesiculosus Protects against Alcohol-Induced Liver Damage by Modulating Inflammatory Mediators in Mice and HepG2 Cells. Mar. Drugs 2015, 13, 1051-1067. https://doi.org/10.3390/md13021051.

Leung, A. Y. (1980). Encyclopedia of Common Natural Ingredients Used In food Drugs and Cosmetics. John Wiley.

Maeda, H., Hosokawa, M., Sashima, T., Funayama, K., & Miyashita, K. (2005). Fucoxanthin from edible seaweed, Undaria pinnatifida, shows an antiobesity effect through UCP1 expression in white adipose tissues. Biochemical and biophysical research communications, 332(2), 392–397. https://doi.org/10.1016/j.bbrc.2005.05.002.

Maliakal, P. P., & Wanwimolruk, S. (2001). Effect of herbal teas on hepatic drug-metabolizing enzymes in rats. The Journal of pharmacy and pharmacology, 53(10), 1323–1329. https://doi.org/10.1211/0022357011777819.

Miłek, M., Marcinčáková, D., Legáth, J. (2019). Polyphenols Content, Antioxidant Activity, and Cytotoxicity Assessment of Taraxacum officinale Extracts Prepared through the Micelle-Mediated Extraction Method. Molecules.24(6):1025.https://doi.org/10.3390/molecules24061025.

Modaresi, M., & Resalatpour, N. (2012). The Effect of Taraxacum officinale Hydroalcoholic Extract on Blood Cells in Mice. Advances in Hematology, 2012.

Muñoz O. Montes M. & Wilkomirsky T. (2001). Plantas medicinales de uso en chile : química y farmacología (1.). Universidad de Chile Vicerrectoría de Asuntos Académicos Comité de Publicaciones Científicas : Editorial Universitaria.

Nair, A. B., & Jacob, S. (2016). A simple practice guide for dose conversion between animals and humans. Journal of basic and clinical pharmacy, 7(2), 27–31. https://doi.org/10.4103/0976-0105.177703.

National Health Services (NHS)- Adios tablets (Dendron Brands Ltd). Last access on 12/12/2023.Avialble on https://dmd-browser.nhsbsa.nhs.uk/amp/view/40210?ref=YW.

Neil Bowdler-BC News (2010). Slimming pills: Do the claims add up? Last access on 12/12/2014.Available on https://www.bbc.com/news/10241981.

Nigatu, T. A., Afework, M., Urga, K., Ergete, W., & Makonnen, E. (2017). Toxicological investigation of acute and chronic treatment with Gnidia stenophylla Gilg root extract on some blood parameters and histopathology of spleen, liver, and kidney in mice. BMC Research Notes, 10(1), 625. https://doi.org/10.1186/s13104-017-2964-3.

Nortadas, R., & Barata, J. (2011). Herbal therapy-induced toxic hepatitis–a case report. Revista da Sociedade Portuguesa de Medicina Interna, 18(2), 89-92.

Olas B. (2022). New Perspectives on the Effect of Dandelion, Its Food Products and Other Preparations on the Cardiovascular System and Its Diseases. Nutrients, 14(7), 1350. https://doi.org/10.3390/nu14071350.

Oliveira Sá, A., Pimentel, T., & Oliveira, N. (2020). Boldo-Induced Hepatotoxicity: A Case of Unexplained Jaundice. European journal of case reports in internal medicine, 7(12), 002116. https://doi.org/10.12890/2020_002116.

Piscaglia, F., Leoni, S., Venturi, A., Graziella, F., Donati, G., & Bolondi, L. (2005). Caution in the use of boldo in herbal laxatives: a case of hepatotoxicity. Scandinavian journal of gastroenterology, 40(2), 236-239.

Pozharitskaya, O. N., Shikov, A. N., Faustova, N. M., Obluchinskaya, E. D., Kosman, V. M., Vuorela, H., & Makarov, V. G. (2018). Pharmacokinetic and Tissue Distribution of Fucoidan from Fucus vesiculosus after Oral Administration to Rats. Marine drugs, 16(4), 132. https://doi.org/10.3390/md16040132.

Rajashekar, V. (2009). Novel nutritional interventions in the manipulation of energy expenditure and blood glucose regulation in humans. PhD thesis University of Westminster School of Life Sciences https://doi.org/10.34737/90z1y.

Ramos-Peralonso, M. J. (2014). European Food Safety Authority (EFSA) Scientific Opinion on Dietary Reference Values for iodine. EFSA J, 12, 1-57. EFSA J, 12, 1–57.https://doi:10.2903/j.efsa.2014.3660.

Relton, C., Li, J., Strong, M. (2014) Deprivation, clubs, and drugs: results of a UK regional population-based cross-sectional study of weight management strategies. BMC Public Health 14, (444). https://doi.org/10.1186/1471-2458-14-444.

Roytman, M. M., Pörzgen, P., Lee, C. L., Huddleston, L., Kuo, T. T., Bryant-Greenwood, P., Wong, L. L., & Tsai, N. (2014). The outbreak of severe hepatitis is linked to the weight-loss supplement OxyELITE Pro. The American journal of gastroenterology, 109(8), 1296–1298. https://doi.org/10.1038/ajg.2014.159.

s, D. I., Chang, A., Viray, M., Chatham-Stephens, K., He, H., Taylor, E., Wong, L. L., Schier, J., Martin, C., Fabricant, D., Salter, M., Lewis, L., & Park, S. Y. (2016). Hepatotoxicity associated with the dietary supplement OxyELITE Pro™ - Hawaii, 2013. Drug testing and analysis,8(3-4),319–327. https://doi.org/10.1002/dta.1894.

Saper, R. B., Eisenberg, D. M., & Phillips, R. S. (2004). Common dietary supplements for weight loss. American family physician, 70(9), 1731–1738.

Schütz, K., Carle,R., & Schieber,A. (2006). Taraxacum—A review on its phytochemical and pharmacological profile. Journal of Ethnopharmacology,107(3), 313-323. https://doi.org/10.1016/j.jep.2006.07.021.

Shin, M. R., Kim, M. J., Park, H. J., Han, J., & Roh, S. S. (2021). Beneficial Effect of Taraxacum coreanum Nakai via the Activation of LKB1-AMPK Signaling Pathway on Obesity. Evidence-based complementary and alternative medicine: eCAM,2021,6655599.

https://doi.org/10.1155/2021/6655599.

Speisky, H., & Cassels, B. K. (1994). Boldo and boldine: an emerging case of natural drug development. PharmacologicalResearch,29(1),1–12. https://doi.org/10.1016/1043-6618(94)80093-6.

Suvarna, K. S., Layton C., and Bancroft J. D.( 2018). Bancroft’s Theory and Practice of Histological Techniques. 8th ed. China: Elsevier Health Sciences. (Chapters 4, 6, 9, 10).

Vodouhè, M.; Marois, J.; Guay, V.; Leblanc, N.; Weisnagel, S.J.; Bilodeau, J.-F.; Jacques, H. (2022) Marginal Impact of Brown Seaweed Ascophyllum nodosum and Fucus vesiculosus Extract on Metabolic and Inflammatory Response in Overweight and Obese Prediabetic Subjects. Mar.Drugs,20,174. https://doi.org/10.3390/md20030174.

Woo, M. N., Jeon, S. M., Shin, Y. C., Lee, M. K., Kang, M. A., & Choi, M. S. (2009). Anti-obese property of fucoxanthin is partly mediated by altering lipid-regulating enzymes and uncoupling proteins of visceral adipose tissue in mice. Molecular nutrition & food research,53(12),1603–1611. https://doi.org/10.1002/mnfr.200900079.

Yang, B., Xie, Y., Guo, M., Rosner, M. H., Yang, H., & Ronco, C. (2018). Nephrotoxicity and Chinese Herbal Medicine. Clinical Journal of the American Society of Nephrology: CJASN,13(10),1605–1611. https://doi.org/10.2215/CJN.11571017.

Zhang, Q., Li, Z., Xu, Z., Niu, X., & Zhang, H. (2003). Effects of fucoidan on chronic renal failure in rats. Planta Medica, 69(6), 537–541. https://doi.org/10.1055/s-2003-40634.

Groups Parameters	Control (T)	Low Dose (LD)	Medium Dose (MD)	High Dose (HD)
WBC(x10³/mm³)	11.55±0.54	11.07±0.89	7.47±0.33	6.60±1.14***
RBC(x10³/mm³)	6.36±1.16	6.04±0.99	6.38±1.86	5.92±1.01
HGB(g/Dl)	13.07±1.74	13.25±0.42	13.30±3.31	13.03±3.16
MCV(Fl)	61.62±0.52	65.30±0.08*	65.10±0.09**	65.17±0.07**
MCHC (g/Dl)	33.40±0.66	33.33±0.44	34.67±0.38*	32.00±0.26
LYM	9.22±0.67	9.00±0.77	6.28±0.93*	5.88±1.01*
RDW-SD(Fl)	30.88±0.41	36.30±0.63*	35.15±0.55*	41.85±0.72***
MPV(Fl)	7.55±3.4	7.88±0.24	7.93±0.01	7.28±1.15
PCT%	0.49±0.01	0.53±0.25	0.57±0.07	0.45±0.22

Groups Parameters	Control (T)	Low Dose (LD)	Medium Dose (MD)	High Dose (HD)
ALB (g/dl)	4.16±0.62	4.14±0.09	4.15±0.12	4.09±0.9
ALP (U/L)	198.20±2.43	169.60±3.8	169.00±3.2	188.40±2.71
ALT (U/L)	55.80±1.34	56.40±1.27	42.00±2.78	46.80±0.93
AST (U/L)	180.60±7.33	168.80±9.28	163.80±4.3	159.00±9.09

Cr (mg/dl)	0.37±0.16	0.39±0.02	0.37±0.13	0.42±0.18
URE (mg/dl)	40.00±2.22	41.80±1.39	43.00±0.03	38.00±0.08

BIOCHEMICAL AND HISTOLOGICAL STUDIES ON ADVERSE EFFECTS OF DIETARY WEIGHT LOSS SUPPLEMENT ON FEMALE RATS: ADIOS