30 facts of Roundup. 30 years of independent research ignored by the assessment agencies.

The most up to date glyphosate review is available here: Sirinathsinghji, 2015.

The two primary ways Roundup/glyphosate enters our daily diet is via: 

  1. Desiccation of our cereals - oats, rye and wheat with Roundup.
  2. Via GMO Roundup Ready food - including, sugar beets, soybeans and canola.

(a) A list of 30 facts of Roundup.  

(b) A list of 20 independent studies not considered for assessment of pesticide safety within the World Health Organisation, the US Environmental Protection Agency or the European Union.

(a) 30 facts of Roundup.  

  1. Maximum residue levels of Roundup / glyphosate are permitted to be higher on our staple cereals (like flour and oats) than on GMO Roundup Ready soybeans & corn.
  2. Pre-harvest pesticide spraying to mature cereal plants & animal feed are the reason glyphosate/ Roundup residues have increased massively on our wheat in the last 6 years.  This is referred to as desiccation or a deathspray.
  3. Residues have increased the same time independent research has been showing damage at increasingly lower levels. Independent research frequently not  included in international toxicity assessments due to the double standards of these assessment agencies.
  4. There are no studies whatsoever held with the major safety agencies of pesticides in their complete form – formula.  Toxicity tests used by the USA EPA, Europe or the WHO never use the stronger formula Roundup, commonly applied to our food. Only the weaker active chemical glyphosate.
  5. In all reproductive & development tests held with the EPA, WHO & EU, rodents arenot dosed the (weaker) glyphosate in the lead up to pregnancy nor in the first 6 days of pregnancy – effectively the first trimester for the rodent.
  6. Independent studies show malformations at the low levels of current human exposure.  These studies are not accepted by the USA EPA, WHO or Europe.
  7. It is rare to find a toxicity study held by the USA EPA, Europe and the WHO for Roundup that is not provided by Monsanto, Cheminova and Syngenta. Or 2,4-D by Dow or the 2,4-D Industry Task Force.
  8. Every unpublished (ie. most of them) Monsanto, Syngenta, Cheminova, Dow (etc,etc,etc) study is unavailable to independent scientists working in the public interest – many problems are dismissed (Eg. due to ‘histopathological data’). These studies are protected by confidentiality agreements between corporation & organisation. These corporate funded studies aren’t published in journals for peer review.
  9. ONLY studies by pesticide corporations end up making the critical studies that declare our safety (the ones that give ‘NOAEL’s’) – Independent studies usually aren’t used.
  10. The 3 major agencies constantly make excuses to keep independent studies out of their glyphosate/Roundup assessments. When independent science declares these same studies credible.
  11. 2 studies with tubular kidney dilation at the same level or lower than the USA permit as residue on wheat are dismissed following a study at a higher glyphosate dose level.  Why?
  12. The glyphosate study that set the Europe ADI dismissed problems at the same level it said was safe.
  13. The study that set the WHO ADI dismissed problems at exactly the same level it said was safe.
  14. In Europe the chemical companies control what studies are put forward for assessing the toxicity of glyphosate/Roundup.  The same companies that have a vested interest in showing low levels of exposure as being safe.
  15. Independent studies indicate glyphosate/Roundup is an endocrine disruptor and potential xenohormone (xenohormones can contribute to breast cancer). References: [a] [b] [c] [d]
  16. Independent studies indicate 2,4-D (the latest pesticide that can be sprayed on GMO products) may contribute towards Parkinsons disease and non-Hodgkins Lymphoma.
  17. In field studies for pesticide related disease in farm workers, results read as ‘inconclusive’ due to the presence of too many pesticides.  However it is almost impossible to finance further work for further testing (including testing for cumulative effects).
  18. GMO products have stacked traits.  You can combine Roundup  2,4-D Ready & glufosinate ‘stacked’ on the same plant. These have never been cumulatively tested for safety.
  19. There is no cumulative testing for Roundup with other pesticides in the USA.  It is excluded because it has a different ‘mechanism of toxicity’ –  effects (Eg. Neurotoxicity) are not considered.
  20. The pesticides used in GMO stacked traits have never been tested together (cumulative testing). There is no study held with the EPA, WHO or EU that has even tried to do this.
  21. The 3 agencies constantly make excuses about problems with the endocrine system by (for example) declaring the dose doesn’t correspond with the response.  The endocrine system is very sensitive and acts differently.
  22. The USA does not test glyphosate/Roundup levels on food  before food is bought at the shops to take home.
  23. Europe is only testing residue levels of glyphosate on cereals. Europe is not testing on other produce.
  24. The studies all held with the EPA, WHO and Europe all show malformations and major problems at the same level that we feed glyphosate (as a residue on stockfeed) to our farm animals.
  25. The USA does not know what its current level of glyphosate exposure is according to the diet of lower socio-economic Americans.
  26. There is no allowance for the more vulnerable needs of developing children.  All the tolerances are based on a 60kg adult.
  27. No study of sperm at current levels of consumption are kept with these organisations.
  28. It was found that the offspring of mothers dosed with glyphosate have lower sperm levels.
  29. No study of sperm is tested using the full formula Roundup.
  30. There has never been further tests with Roundup and the thyroid gland.  All tests (available on the sites) with glyphosate and the thyroid gland showed tumour growth.

(b) 30 years of independent studies not funded by pesticide corporations.

1985. Scientists show that retinoic acid [1] (essential in early developmental growth) can be teratogenic [2] to humans (like other mammals). [3]

1989. Laboratory studies show teratogenic effects of retinoic acid in the central nervous system of animals. [4]

1992. This study found that, irrespective of the dose, route or frequency of duration 1.4% of the administered dose (of glyphosate) was retained in tissues. The highest concentration of radioactivity was present in bone, with lower concentrations in bone marrow, kidney, liver, lungs and the residual carcass. (This study is considered by the WHO.)  [5]

1995.  Glyphosate on NZ white rabbits. Pesticide treatment resulted in a decline in body weight, libido, ejaculate volume, sperm concentration, semen initial fructose and semen osmolality. This was accompanied with increases in the abnormal and dead sperm and semen methylene blue reduction time. The hazardous effect of these pesticides on semen quality continued during the recovery period, and was dose-dependent.  [6]

1997. A study recommended the possibility that the increasing use of certain plastics and insecticides that degrade into substances that have estrogen-related physiological effects on living things should be investigated in relation to the earlier onset of puberty.  [7]

2000. Study finds Roundup formulation interferes with an enzyme involved in the production of testosterone in mouse cell culture.  While glyphosate did not alter steroidogenesis (progesterone production) or total protein synthesis at any dose tested (0–100g/ml), the glyphosate formulation Roundup decreased progesterone production in a dosage-dependant manner without a parallel decrease in total protein synthesis. [8]

2001. Glyphosate found to induce a variety of abnormalities in foetuses and pregnant rats. [9]

2002.  In the submission to Europe for assessment of glyphosate, many studies found heart malformations. Many of these findings were dismissed.   As with the liver, gonads and kidneys, the heart is a soft organ that fulfils a secondary endocrine function.

2004.  Retinoic acid signaling is confirmed as essential for pancreas development and promotes endocrine at the expense of exocrine cell differentiation in Xenopus. [10]

2005. Glyphosate found to inhibit aromatase activity (important enzyme, and a key step in the biosynthesis of estrogens) at dilutions 100 lower than agricultural rates – and the Roundup formulation produced greater effects than glyphosate alone.  The study authors suggest that the presence of Roundup adjuvants enhances glyphosate bioavailability and/or bioaccumulation [11]

Aromatase enzyme can be found in many tissues including gonads, brain, adipose tissue, placenta, blood vessesls, skin, bone and endometrium.   In particular, aromatase is responsible for the aromatization of androgens into estrogens is an important factor in sexual development.  (Wikipedia)

2007. Roundup confirmed to be an aromatase disruptor (disrupting conversion of steroids to estradiol).  Study reveals that the cytotoxic and potentially endocrine disrupting effects of Roundup are amplified with time.  Study suggests that Roundup exposure may affect human reproduction in case of contamination, and that chemical mixtures in formulations appear to be underestimated regarding their toxic or hormonal impact.  [12]

2007. Dallegrave examined the reproductive effects of Roundup on male and female offspring of Wistar rats treated with 50, 150 or 450 mg/kg of Roundup during pregnancy and lactation. The study found that these doses of Roundup did not induce maternal toxicity but did induce adverse reproductive effects on male offspring. Findings include a decrease in sperm number and daily sperm production during adulthood, an increase in the percentage of abnormal sperms, a dose-related decrease in the serum testosterone level at puberty, and signs of sperm cell degeneration during both periods. The study showed that Roundup is a reproductive toxin at non-maternally toxic doses.  [13]

1999-2008: Cancers with increasing incidence trends for the USA include cancers of the pancreas, liver, thyroid, and kidney.  [14]

2008. Over 80% of the current GM crop acreage worldwide is planted in 4 herbicide tolerant crops, the vast majority are tolerant to high levels of glyphosate.  The crops are soybean, maize, canola and cotton, grown mainly in USA, Canada, Argentina, Brazil, and Paraguay.  [15]

2008. Profesor Carrasco (Argentina) states glyphosate is not breaking down in cells, but rather, accumulating.  The findings lend weight to the claims that abnormally high levels of cancer, birth defects, neonatal mortality, lupus, kidney disease and skin and respiratory problems in populations near Argentina’s soybean fields may be linked to the aerial spraying of Roundup. [16]

2009.  At dilution level far below agricultural recommendations and corresponding to low levels of pesticide residues in food or feed, independent scientists Benchour and Seralini concluded that Roundup adjuvants like POEA change human cell permeability and amplify toxicity induced already by glyphosate, through apoptosis and necrosis.  Three human cell lines were tested from  10ppm to 2% (ag usage level):the primary cell line HUVEC from umbilical cord vein epithelium, embryonic cell line 293 derived from kidney, and placenta cell line JEG3.  All cells died within 24 hours of exposure to the Roundup formulations. Creating evidence that the mixes available on the market cause cell damage and even death around the residual levels currently expected from Roundup treated crops. [17]

2009.  This analysis highlights that the there was a clear negative impact on the function of kidney and liver organs in rats consuming GM maize varieties for just 90 days. Other effects were also noticed in the heart, adrenal glands, spleen and haematopoietic system. The authors concluded that these data highlight signs of hepatorenal toxicity, possibly due to the new pesticides specific to each GM corn. In addition, unintended direct or indirect metabolic consequences of the genetic modification cannot be excluded. [18]

2009.  At low doses corresponding with accepted residue levels in food today, scientist at the University of Caen confirmed human cell lines find glyphosate based herbicides to be toxic and to be endocrine disruptors.  At 0.5 ppm endocrine cells were disrupted, then from 2 ppm the transcriptional activities on both estrogen receptors were inhibited. Aromatase transcription and activity were disrupted from 10 ppm. Cytotoxic effects started at 10 ppm ,  and DNA damages at 5 ppm. The Caen team suggested  “real cell impact of glyphosate-based herbicides residues in food, feed or in the environment has thus to be considered, and their classifications as carcinogens/mutagens/reprotoxics” discussed. [19]

2010. Carrasco finds that glyphosate increases retinoic activity in frog embryos and reveals the mechanism as to how malformations have occurred.  Frog and chicken embryos fed low doses of Roundup mix (lower than ag spraying) were found to produce teratogenic effects (malformations) by impairing retinoic acid signalling.  The study concludes that glyphosate effects morphogenesis:  the biological process that causes an organism to develop its shape.  [20]

NB: Within months of the release of the Carrasco study EU regulators introduced a delay on new legislation that would have ensured independent research would be included in the EU 2012 toxicological assessment of glyphosate – they delayed the legislation coming in until 2015 which meant glyphosate would be reviewed only according to what the agrichemical companies wanted to put forward in their studies for reassessment.  This would result in old lax standards of assessment.  Glyphosate is only reviewed every 15 years.  For the full story please read the Open Source document Roundup and Birth Defects.

2010.  All the four Roundup formulations in this study provoked liver cell death, with adjuvants having stronger effects than glyphosate alone.  A plant extract, Dig 1, non cytotoxic and not inducing caspases by itself, is able to prevent Roundup-induced cell death in a time-dependant manner. [21]

2010.  This study demonstrated a significant reduction in serum testosterone concentrations and changes in the testicular morphology of male Wistar rats treated with Roundup during puberty, and these effects were demonstrated with even the lowest dose used in the experiment (5 mg/kg).. [22]

2012 No studies to date have experimentally examined the causal relationship between exposure to pesticides directly from conventionally grown foods and adverse neurodevelopmental health outcomes.  [23]

2012.  This study only dosed glyphosate to the mothers/dams – but the offspring females had increased estradiol serum concentrations and earlier puberty; the males also had increased estradiol serum concentrations but sexual arousal, which estradiol is meant to stimulate, but this didn’t happen. [24]

Estradiol has a critical impact on reproductive and sexual functioning.   Additionally, if testes are exposed to either too high or too low levels of estrogens (such as estradiol) spermatogenesis can be disrupted to such an extent that the animals become infertile. (Wikipedia)

2012.    The  lifetime feeding study that reveals tumours in rats.  The results of the study presented here clearly demonstrate that lower levels of complete agricultural glyphosate herbicide formulations, at concentrations well below officially set safety limits, induce severe hormone-dependent mammary, hepatic and kidney disturbances.  The study also questions the contribution of GMO organisms [25]  More reading on the Seralini site and on the terrific, science backed GMwatch site.

Monsanto, Cheminova and Syngenta etc. have never been interested in conducting lifetime feeding studies at low levels of the glyphosate formulation (including Roundup) that we are exposed to on a daily basis.  Go on we dare you – find one that is used in critical assessments with the WHO, EU or US EPA - and email RITE.  Find 3.  And throw in a generational study for good measure. But you need to show us the full study.  Because right now, their studies are the only studies held with the world agencies that decide our pesticide exposure levels. And they don’t look at the glyphosate mix at these very low levels.  That is why the Seralini study is so important.  

2012  A study examining the teratogenic effects of glyphosate based herbicides concluded [26]:

A substantial body of evidence demonstrates that glyphosate and Roundup cause teratogenic effects and other toxic effects on reproduction, as well as genotoxic effects. From an objective scientific standpoint, attempts by industry and government regulatory bodies to dismiss this research are unconvincing and work against the principle that it is the responsibility of industry to prove that its products are safe and not the responsibility of the public to prove that they are unsafe. The precautionary principle would suggest that glyphosate and its commercial formulations should undergo a new risk assessment, taking full account of the entirety of the peer-reviewed scientific literature as well as the industry-sponsored studies. Experience to date suggests that the new risk assessment should be conducted with full public transparency by scientists who are independent of industry.

2013  This study demonstrated that at low and environmentally relevant concentrations (Ie. much lower than the all studies held with the US EPA, WHO & EU) of glyphosate possessed estrogenic activity – it had the potential to be a xenohormone and endocrine disruptor. In this study glyphosate exerted proliferative effects (via the estrogen receptor) in human hormone-dependent breast cancer, T47D cells [27]

An indepth review of the toxic effects of the herbicide, glyphosate, the active ingredient in Roundup, in humans, and demonstrated how glyphosate’s adverse effects on the gut microbiota, in conjunction with its established ability to inhibit the activity of cytochrome P450 enzymes, and its likely impairment of sulfate transport, can explain a great number of the diseases and conditions that are prevalent in the modern industrialized world.   [28]

2014  A study making a comparison of the genotoxicity of glyphosate demonstrated that genotoxic in the cells and organisms studied at concentrations of 0.7–7 μM. [29]

Despite its relatively benign reputation, Roundup was among the most toxic herbicides and insecticides tested in another study. Most importantly, 8 formulations out of 9 were up to one thousand times more toxic than their active principles. Green Med Info reported that this revealed Roundup to be 125 times more toxic than regulators advised. [30]

In a recent study researchers conducted a systematic review and a series of meta-analyses of nearly three decades worth of epidemiologic research on the relationship between non-Hodgkin lymphoma (NHL) and occupational exposure to agricultural pesticide active ingredients and chemical groups. Random effects meta-analyses showed that phenoxy herbicides, carbamate insecticides, organophosphorus insecticides and the active ingredient lindane, an organochlorine insecticide, were positively associated with NHL.  This review indicated the need for investigations of a larger variety of pesticides in more geographic areas, especially in low- and middle-income countries, which, despite producing a large portion of the world’s agriculture, were missing in the literature that were reviewed. [31]

 

Return to top

References:

[1] Retinoic acid is a metabolite of vitamin A (retinol) that mediates the functions of vitamin A required for growth and development. Retinoic acid is required in chordate animals, which includes all higher animals from fish to humans. Wikipedia.

[2] Teratogens are substances or environmental agents which cause the development of abnormal cell masses during fetal growth, resulting in physical defects in the foetus. Wikipedia

[3]   Edward J. Lammer, M.D., Diane T. Chen, M.D., M.P.H., Richard M. Hoar, Ph.D., Narsingh D. Agnish, Ph.D., Paul J. Benke, M.D., Ph.D., John T. Braun, M.D., Cynthia J. Curry, M.D., Paul M. Fernhoff, M.D., Art W. Grix, Jr., M.D., Ira T. Lott, M.D., James M. Richard, M.D., and Shyan C. Sun, M.D. Retinoic Acid Embryopathy.    N Engl J Med 1985; 313:837-841October 3, 1985DOI: 10.1056/NEJM198510033131401

[4]  Durston, A. J.; Timmermans, J. P. et al. (13). “Retinoic acid causes an anteroposterior transformation in the developing central nervous system”Nature 340, 1989 (6229): 140–144.doi:10.1038/340140a0

[5] Powles, P. (1992b) 14C-glyphosate: absorption, distribution, metabolism, and excretion in the rat. Unpublished report No. 7006-676/2, dated 30 June 1992, from Hazleton UK, Harrogate, England. Submitted to WHO by Cheminova A/S, Lemvig, Denmark.

[6]  Yousef MI, Salem MH, Ibrahim HZ, Helmi S, Seehy MA, Bertheussen K. J..  Toxic effects of carbofuran and glyphosate on semen characteristics in rabbits.   Environ Sci Health B. 1995 Jul;30(4):513-34.

[7]  Herman-Giddens, et al. 1997. “Secondary sexual characteristics and menses in young girls seen in office practice: a study from the pediatric research in office settings network.” Pediatrics 99(4):505-512.

[8]  Walsh LP, McCormick C, Martin C, Stocco DM. “Roundup inhibits steroidogenesis by disrupting steroidogenic acute regulatory (StAR) protein expression.” Environ Health Perspect. 2000 108: 769–776.

[9] Daruich J, Zirulnik F, Gimenez MS (March 2001). “Effect of the herbicide glyphosate on enzymatic activity in pregnant rats and their fetuses”Environmental Research 85 (3): 226–31.

[10] Chen Y, Pan FC, Brandes N, Afelik S, Solter M, et al. (2004) Retinoic acid signaling is essential for pancreas development and promotes endocrine at the expense of exocrine cell differentiation in Xenopus. Dev Biol 271: 144–160.

[11]  Richard, Sophie; Moslemi, Safa; Sipahutar, Herbert; Benachour, Nora; Seralini, Gilles-Eric (2005). “Differential Effects of Glyphosate and Roundup on Human Placental Cells and Aromatase”.  Environmental Health Perspectives 113 (6): 716–20. doi:10.1289/ehp.7728.

[12]  Benachour, N., Sipahutar, H., Moslemi, S., Gasnier, C., Travert, C., Séralini, G-E. 2007. Time- and dose-dependent effects of roundup on human embryonic and placental cells. Archives of Environmental  Contamination and Toxicology 53, 126–33.

[13]  Dallegrave, E., Mantese, F. D. et al. 2007. Pre- and postnatal toxicity of the commercial glyphosate formulation in Wistar rats. Arch Toxicol 81: 665–673.

[14] Edgar P. Simard PhD, MPH1,*, Elizabeth M. Ward PhD2, Rebecca Siegel MPH3, Ahmedin Jemal DVM, PhD4   Cancers with increasing incidence trends in the United States: 1999 through 2008 first published online: 4 JAN 2012 DOI: 10.3322/caac.20141

[15] Villar JL, Freese B. 2008.  Who Benefits From GM Crops?  The rise in Pesticide Use. Friends of the Earth International, Amsterdam. .

[16]  Paganelli, A., Gnazzo, V. et al. 2010. Glyphosate-based herbicides produce teratogenic effects on vertebrates by impairing retinoic acid signaling. Chem Res Toxicol 23(10): 1586–1595.

[17]   Benachour, N., Séralini, G-E. 2009. Glyphosate formulations induce apoptosis and necrosis in human umbilical, embryonic, and placental cells.  Chem. Res. Toxicol. 22, 97–105.

[18]  de Vendômois JS, Roullier F, Cellier D, Séralini GE.  A comparison of the effects of three GM corn varieties on mammalian health.  Int J Biol Sci. 2009 Dec 10;5(7):706-26.

[19]   Gasnier C, Dumont C, Benachour N, Clair E, Chagnon MC, Séralini GE.  Glyphosate-based herbicides are toxic and endocrine disruptors in human cell lines. Toxicology. 2009 Aug 21;262(3):184-91. Epub 2009 Jun 17.

[20]   Paganelli, Alejandra; et. al. “Glyphosate-Based Herbicides Produce Teratogenic Effects on Vertebrates by Impairing Retinoic Acid Signaling”.   Chem. Res. Toxicol. 23 (10): 1586 1595. doi:10.1021/tx1001749. PMID 20695457.    http://pubs.acs.org/doi/abs/10.1021/tx1001749

[21]   Gasnier C, Benachour N, Clair E, Travert C, Langlois F, Laurant C, Decroix-Laporte C, Séralini GE. Dig1 protects against cell death provoked by glyphosate-based herbicides in human liver cell lines.  Source: Laboratory of Biochemistry EA2608, Institute of Biology, University of Caen, France. criigen@unicaen.fr.

[22]    R.M. Romano, M.A. Romano, M.M. Bernardi, P.V. Furtado, C.A. Oliveira.  Prepubertal exposure to commercial formulation of the herbicide glyphosate alters testosterone levels and testicular morphology. Arch. Toxicol., 84 (2010), pp. 309–317

[23] American Academy of Paediatrics,   Organic Foods: Health and Environmental Advantages and Disadvantages. doi: 10.1542/peds.2012-2579

[24]  Romano MA, et al.  Glyphosate impairs male offspring reproductive development by disrupting gonadotropin expression.   Arch Toxicol. 2012 Apr;86(4):663-73. doi: 10.1007/s00204-011-0788-9. Epub 2011 Nov 26.

[25]  Séralini, GE, Clair E, Mesnage R, Gress S, Defarge N, Malatesta M, Hennequin D and de Vendômois JS.  Republished study: long-term toxicity of a Roundup herbicide and a Roundup-tolerant genetically modified maize. RESEARCH Open Access Springer. Environmental Sciences Europe 2014, 26:14.

[26]  Antoniou M, Habib MEM, Howard CV, Jennings RC, Leifert C, et al. (2012)Teratogenic Effects of Glyphosate-Based Herbicides: Divergence of Regulatory Decisions from Scientific Evidence. J Environ Anal Toxicol S4:006. doi:10.4172/2161-0525.S4-006

[27]  Glyphosate induces human breast cancer cells growth via estrogen receptors. Thongprakaisang S1, Thiantanawat A, Rangkadilok N, Suriyo T, Satayavivad J. Food Chem Toxicol. 2013 Sep;59:129-36. doi: 10.1016/j.fct.2013.05.057. Epub 2013 Jun 10.

[28]  Glyphosate’s Suppression of Cytochrome P450 Enzymes and Amino Acid Biosynthesis by the Gut Microbiome: Pathways to Modern Diseases. Anthony Samsel and Stephanie Seneff.  Entropy 2013, 15(4), 1416-1463; doi:10.3390/e15041416

[29]  Comparison of the in vivo and in vitro genotoxicity of glyphosate isopropylamine salt in three different organisms.  Carlos Alvarez-Moya, Mónica Reynoso Silva, Carlos Valdez Ramírez, David Gómez Gallardo, Rafael León Sánchez,Alejandro Canales Aguirre, and Alfredo Feria Velasco. Genet Mol Biol. Mar 2014; 37(1): 105–110.  Published online Feb 28, 2013.  PMCID: PMC3958316

[30] Major Pesticides Are More Toxic to Human Cells Than Their Declared Active Principles.  Mesnage, R.; Defarge, N.; Spiroux de Vendômois, J.; Séralini, G. E.  BioMed Research International, 2014 :179691. doi: 10.1155/2014/179691. Epub 2014 Feb 26.

[31]  Non-hodgkin lymphoma and occupational exposure to agricultural pesticide chemical groups and active ingredients: a systematic review and meta-analysis. Schinasi, L.; Leon, M,E.  International Journal of Environmental Research and Public Health, 2014, 23;11(4):4449-527. doi: 10.3390/ijerph110404449.