Grape Skin Extraction (GSE) increases life span of Parkinson’s disease model of Drosophila by promotion of Autophagy.Abstract: Studies have shown that Red wine has benefits after ingesting to one’s health examples of this include increasing women’s fertility, cancer, age related neurodegenerative diseases such as Parkinson’s disease. These benefits are said to have come from nutrients within grape skin known as resveratrol, which is an antioxidant and has proven to prolong life-span. Grape Skin extraction was added to Drosophila daily food intake to observe the benefits of this new supplement. The impact of grape skin extraction on Drosophila Loss of PINK1 gene was studied by looking at the models life span and the promotion of autophagy. This loss of gene allowed the model to be a representation of Parkinson’s disease. The results present the association between the promotion of autophagy and benefits of Grape skin Extraction. Results also presented the protection of mitochondrial function which helps the prevention of the formation of Parkinson’s disease.1. IntroductionThe oldest known winery was dated in 4100 BC and red wine specifically has been known to have health benefits after consumption. After scientific studies it has been known for Red wine to be advantageous for women’s fertility, people with cardiovascular diseases and neurodegenerative diseases such as Parkinson’s disease and Alzheimer’s disease (Telegraph reporters et al., 2017) The known compound found in wine which contributes to such affects is known as resveratrol, which acts as an antioxidant, defends against the clustering of neurotoxic protein and involved in anti-inflammatory responses. (P J.Skerret et al., 2012)Listed neurodegenerative diseases of which resveratrol is a benefit of are very common, however Alzheimer’s is the most present disease before Parkinson’s disease. There are symptoms that involve stiffness of limbs, Tremor, impaired balance and dementia all of which can result in needing assistance for daily life tasks (Heyn, Patrick Davis, Convad Stoppler et al 2018). There are a recorded number of 10 million people worldwide that have Parkinson’s disease. It is thought that Parkinson’s disease cases increase with age, commonly beginning at the age of over 50; men being more probable to have the disease than women (Clinaero, et al., 2006-2018). To this current day there is no antidote for Parkinson’s disease or Alzheimer’s, however there are medications or diets that are known to supress the symptoms of both diseases. (nhs.uk. 2018) For example adapting to a Mediterranean diet, this involves a fruit and vegetable based diet, low in meat, saturated fats, sugar and moderate intake of dairy products, oily fish and small quantity of red wine during meals. The known benefit from this diet was the attribute of consuming red wine, specifically the content of concentrated Polyphenols obtained from grape skin during the fermentation process (Alzheimer’s Society. 2018). The grape skin that is used in the manufacturing of red wine consist of anti-oxidative components e.g. proanthocyanide, quarcetin. They are all ingredients which are active in providing a shield for the presence of free-radical mediated tissue injury and oxidative stress. (F.Cosme, A. Vilela, T. Pinto (2018) Phenolic compounds and antioxidant activity in grape juices: a chemical and sensory view, beverages 4, 22)The diagnosis of Parkinson’s disease is deciphered by considerations of genetic backgrounds and environmental factors. PINK1 is a gene that defends cells from stress induced mitochondrial dysfunction by inducing autophagy of the mitochondria (Narendra DP et al., 2010) PARKIN is a gene that is involved in the degradation of unnecessary proteins by tagging impaired and surplus proteins with molecules called ubiquitin. (U.S. National Library of Medicine et al 2018). Both PINK1 and PARKIN are connected to autosomal recessive forms of Parkinson’s disease. (Valente et al., 2004; Kitada et al., 1998) With this being said they have visible differences when it comes to what function their genes encrypt. For example PINK1 gene at the N-terminus with a mitochondrial-targeting signal encrypts a serine-threonine kinase. This gene is involved in the monitoring of mitochondrial quality control by presenting the outer membrane of the mitochondria linked with translation, revealing damage in this area; whereas PARKIN gene encrypts an E3 ubiquitin ligase. The presence of this gene when the mitochondria are damaged advances a process that involves degradation of the mitochondria. (Narendra et al., 2008; Narendra et al., 2010; Vives-Bauza et al., 2010; Matsuda et al., 2010; Gehrke et al., 2015) In drosophila Melanogaster a deficit of PINK1 gene leads to the deterioration of neurones that release dopamine and indirect flight muscles along with affecting the duration of the life spam by decreasing it. This known physiology process helps guide the role of the impaired mitochondrial in model with developing Parkinson’s disease (Park et al., 2006; Clark et al., 2006; Yang et al., 2006).The grape skin extraction is investigated using Drosophila PINK1 model for the security against the damaging effects of impaired mitochondria. This was done by simply adding Grape seed extraction powder into their daily dose of food that was given to the Drosophila. Observations were made and benefiting developments were seen in their health, such as the flight muscle function. (Z.Wu, A.Wu, Dong, Sigears and Lu et al., 2018)2. DiscussionThere are current developments which look to prevent Parkinson’s disease, such treatment involves trying to increase the secretion of dopamine from neurotransmitter and stop the degradation of dopamine (Kim SW et al 2005). To this present day there is no antidote for the initiation of such neurodegenerative diseases. There are reports in which state that there are health benefits from the consumption of red wine, green tea, however it is still unclear as to why they present such benefits (Morgan and Grundmann, 2017). The data presented clearly states that Grape Skin Extraction demonstrates good benefits in an animal model with Parkinson’s disease. It is later displayed that grape skin extraction could assist in sustaining mitochondrial function by induction of Autophagy which eliminates dysfunctional or impaired mitochondria in PINK1 loss-of function Parkinson’s disease flies. There are some differences of results with regards the benefits of Grape skin extraction as other reporters have stated that it defends mitochondria from oxidative damage in Parkinson’s disease model using Drosophila (Long et al., 2009)Such reports state that whole grapes contain a particular extract known as botanical extract this substance consist of several micro-nutrients such as flavans, anthocyanins, resveratrol and emodin. All of which possess a function that is anti-oxidative.  They protect the activity of respiration in the mitochondria, including both the electron transport chain (complex I and II) and pyruvate dehydrogenase.  Such extract was given to a Drosophila of which had a gene transferred (human ?-synuclein). This was then a model Parkinson’s disease with malfunctioning movement, loss of neurotransmitters that release dopamine. This was presented by the inability to climb using geotaxis (the response of a free moving organism to gravity procedures (Miller-Keane et al., 2003)). The Drosophila presented noticeable advancements in both lifespan and ability to climb (Long, Gao, Sun, Lui and Zhao-Wilson et al., 2009).Figure 1. Analysis ofPINK1 mutant fly health-span after feeding GSE or resveratrol-supplemented food. Parameters measured are jump/flight ability (C) ATP Production (D) (Z.Wu, A.Wu, Dong, Sigears and Lu et al., 2018)During the life-span study shown in figure 1 it was presented by PINK1 mutants of which showed a shorter life span than wild type flies, this is the same result as previous reports (Yang et al., 2006). There was an increase in the median life span of PINK1 mutant flies when such flies consumed 4% and 8% concentrations to normal fly food. However no such observations were seen with the wild type flies under the same conditions. The higher dosage of 16% concentration of GSE showed no significant change in the median life span of wild type of PINK1 mutant, however displayed an improved effect on the pathological and biochemical processes. It is obvious that lifespan is a very complicated phenotype and is related to many environmental and genetic factors, which makes the overall observation and finding very complex task compared to other phenotypes. When it comes to providing a high concentration of resveratrol proved only to change the lifespan of the PINK1 mutant and not the wild type fly. Demonstrating that PINK1 mutant flies is responsive to GSE supplement. It has been reported through other research that resveratrol has beneficial effects on other organisms such as its lifetime durations for example Nematodes and Yeast (Wood et al., 2004). But this benefit is proven debateable upon other findings due to the non-existing advantageous effects. (Bass et al., 2007). Another controversy was shown when it was presented that red and white wine without any resveratrol substances still had beneficial effects on the life span by stimulating cell survival. (Mukherjee et al., 2009) This shows that resveratrol within grape skin extraction alone, may not be the factor which is giving these effects on the life-span.Figure 2 A supplement of GSE suppressed the abnormal wing posture phenotype and improved Health-span in mutant flies. (A) Known volumes of dyed food consumption by PINK1 mutant flies with indicated intervals of dosages. (B) Usual abnormal wing positioning of PINK1 mutant flies, known volumes of GSE supplement was raised with varied concentration after 7 days of aging at 250C * or # , p<0.05 compared to NF in 7 days or 15 days records. (Z.Wu, A.Wu, Dong, Sigears and Lu et al., 2018)As shown in Figure 2 it is apparent that mutant flies with PINK1 presented the breaking down of indirect muscle post and present to the change of appearance by the visible abnormality in the positioning of the wing. Wild type flies physical traits show the wings lying in line with the angle of their body, whereas PINK1 mutant male and female flies show a wilted or upright wing posture.Activation of PINK1 gene is suggested to result in increases request for energy in order to carry out the function of mitochondria and sustain the condition of the mitochondria. However the non-activated PINK1 presents aggregation of mitochondria with unusual anatomy. (Liu and Lu, 2010: Wu et al., 2013). When considering the fluorescent imaging as presented in Figure 3 the PINK1 mutant flies presented a mass cluster of mitochondria, and Wild type showed the standard anatomy of mitochondria with similar sizes. High measures of Grape extraction being 8% and 16% resulted in low quantities of clustering mitochondria thus showing the removal of high levels of aggregation.  Figure 3. The supplement of GSE partially removed the accumulation of mitochondria in in the flight muscle of PINK1 RNAi flies. Mitochondria are observed and measured using Mitochondrial- targeted GFP (mito-GFP). The areas with dense quantities of green indicate accumulation of mitochondria in flight muscle. Scale bar, 50??m. (B) Quantification of data shown in A. Mitochondrial aggregates larger than 5??m and 10??m were scored in a 200??m?×?200??m region from 3–4 independent areas. * or #, p?5??m or 10??m diameters. (Z.Wu, A.Wu, Dong, Sigears and Lu et al., 2018)Autophagy is a process that involves the self-breakdown of damaged organelles within a cell, playing a vital role in response to the stress of inadequate nutrition of total nutrients and for stabilising foundations of energy at important times. (Glick and Barth and Macleod et al., 2010) Faults in autophagy are linked with neurodegeneration, cancer and aging (Mizushima et al., 2008) PINK1 and PARKIN have been associated with the activation of the mitochondrial autophagy pathway and are involved in the regulation of the quality control in mitochondria (Yang and Lu, 2009; Durcan and Fon, 2015). This particular data makes certain of the decrease of aberrant mitochondria in PINK1 mutant flies after consumption of Grape Skin extraction compared to the wild type Drosophila. (Z.Wu, A.Wu, Dong, Sigears and Lu et al., 2018)3. ConclusionWith this all taken into consideration this study implies the effect of resveratrol in autophagy induction is not as compatible as gene transfer extraction, however altering the chemical preparation of mitochondrial pathways when promoting autophagy process by using grape skin extraction supplement may suggest a positive advancement to battle against Parkinson’s disease. When using high percentage concentration of resveratrol the life-span in PINK1 mutant flies was noticed and had the anticipated reaction. The overall implications from this study suggest that the supplement of resveratrol of life span on PINK1 mutant flies originate from the effect on the other visible features of mitochondrial function not specifically autophagy. Grape skin’s nutrients include quercetin, myricetin, catechins, tannis and resveratrol, therefore being possible that the non-resveratrol components may enhance Autophagy. A similar study showed that Amurensin G, compound from wild grape could induce autophagy and attenuate cellular toxicities in a rotenone model of Parkinson’s disease in human cells (Ryu et al., 2013).Results from this study correlate with the results of previous reporters in that grape skin extractions contains nutrients which are beneficial to those with Parkinson’s disease. Furthermore Grape skin extraction applied its protective effects by enhancing Autophagy. So in the future a combination of nutrients from Grape skin extraction and medication could be used as a therapeutic potential for the neurodegenerative disease. Though, before testing on humans it would be essential to continue studies on models of Parkinson’s disease in other species.ReferencesMedicineNet. 2018. Parkinson’s Disease Early and Later Symptoms, 5 Stages, and Prognosis. [ONLINE] Available at: https://www.medicinenet.com/parkinsons_disease/article.htm#what_are_the_early_signs_and_symptoms_of_parkinsons_disease. [Accessed 28 October 2018].The Telegraph. 2018. Red wine: the unexpected health benefits. [ONLINE] Available at: https://www.telegraph.co.uk/health-fitness/nutrition/red-wine-unexpected-health-benefits/. [Accessed 28 October 2018].Patrick J.Skerret (2012) Harvard Publishing, Harvard medical schoolSietske N.Heyne, C.Patrick Davis, M. 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