CHAPTER ONE
1.1 INTRODUCTION
Cola Nitida has been used in folk medicine as an aphrodisiac, an appetite suppressant, to treat morning sickness, migraine headache, and indigestion (Esimone et al., 2007). It has also been applied directly to the skin to treat wounds and inflammation (Newall et al., 1996)on the teeth and gums. There has also been speculations as to its use in the treatment of peptic ulcer, however, in Nigeria there is no record of the incidence of peptic ulcer.
According to Esimone (2007), cola nitida tree is native to West Africa. Cola nuts are obtained from cola trees. Cola nitida belongs to the genus cola and family steriliaceae. They are commonly used to counteract hunger and thirst; in some cases it is used to control vomiting in pregnant woman and also as a principal stimulant to keep awake and withstand fatigue by students, drivers, and other menial workers.
Lipoproteins are found in the liver and cells. The liver is the largest organ inside the body. The liver is dark reddish-brown and
consists of two main lobes. There are over 300 billion specialized cells in the liver that are connected by a well organized system of “bile” ducts and blood vessels called the biliary system.
THE OXIDATIVE STRESS STATUS OF RATS FED ON OIL BEAN SEED
1.0. INTRODUCTION
Ugba also called ukpaka is a popular food delicacy in Nigeria especially among Igbo ethnic group. It is rich in protein and is obtained by a solid state fermentation of the seed of African oil bean tree (Pentaclethra macrophylla Benth). The natural fermentation of the seed which at present is still done at the house-hold level, renders the production nutritious, palatable and non-toxic (Enujiugha, 2002).
Its production, like many African fermented foods depends, entirely on mixed fermentation by microorganism from diverse source.
Pentaclethra macrophylla Benth is a large woody plant abundant in the rain forest areas of west and central Africa. It’s origin in Nigeria is believed to be around 1937 (Ladipo, 1984); where it is found in the South Nigeria, (Mbajunwa et al., 1998).
―Ugba‖ Pentaclethra macrophylla Benth belongs to the Family Leguminosae and sub-family Microsoideae (Keay, 1989 and NFTA, 1995).
Ugba seeds are irregular and oval; they are flat, black and hard pods. It is composed of oil, protein and small amounts of carbohydrate (Obeta, 1982).
1.1 AIM AND OBJECTIVES
(i) To determine the concentration of Malondialdehyde (MDA, which indicates the peroxidation status) and
(ii) The activity of Catalase (a marker of antioxidant status) in the serum of rats fed graded doses of African oil bean seed meal
REDUCTION OF YOGURT FROM SOYMILK AND COW MILK USING PINEAPPLE (ANANASCOMOSUS) AS SWEETENERS
CHAPTER ONE
1.0 INTRODUCTION
Interest in functional foods has recently increased among consumers due to a greater consciousness of health and nutrition; as well as the need to cure diseases and also the increasing scientific evidence of their effectiveness. Fermented products are a significant part of many indigenous diets.
Yoghurt is a Turkish name for a fermented milk product. It is originated by early normadic herdsman, especially in Asia, Southern and Eastern Europe. Yoghurt is made by adding a culture of acid forming bacteria to milk that is usually homogenized, pasteurized and fermented. Yoghurt is defined as a fermented milk product that evolved empirically some centuries ago by allowing naturally contaminated milk to sour at a warm temperature, in the range of 40-50 °C (Kwon et al., 2011). The micro-organisms which are used conventionally in this process are referred to as “Starter Culture”. They includeLactobacillus delbrueckii subsp. Bulgaricus and Streptococcus thermophilus. The average size ofLactobacillus bulgaricus ranges from 0.8 to 1.0μm in diameter (Rakis .,1978 , Sanfu. , 2009).
During the fermentation, hydrolysis of the milk proteins occurs, the pH drops, the viscosity increases and bacterial metabolites are produced that contribute to the taste and possibly to the health promoting properties of yoghurt. The sugars are fermented by the bacteria into lactic acid, which causes the formation of the characteristic curd. The acid lowers the pH of the yoghurt and restricts the growth of food poisoning bacteria (putrefactive or pathogenic).
Not only is yoghurt a wonderful quick, easy and nutritious snack, but also research evidence point to the fact that milk and yoghurt may actually add years to life as found in some countries where fermented dairy products are a dietary (Wolf.,1978) Several health benefits have been reported for traditional yoghurt and this healthy image is enhanced by supplementation with probiotic bacteria ( ; Magenis, et al 2006).
Soymilk is an aqueous extract of soya beans (Glycine max) and is quiet similar in appearance to cow milk (Agure-Dam, 1997). It is commonly characterized as having a beany, grassy or soy flavor, which reportedly can be improved by lactic acid fermentation, as in yoghurt-like products Microorganisms possess endogenous β-glucosidases which can be utilized to hydrolyze predominant isoflavone glucosides in soymilk to improve biological activity.
It has been reported that probiotic organisms including Bifidobacteria and some other lactic acid bacteria hydrolyze isoflavone glycosides into corresponding aglycones (Cheng., 1998).
Recent reports indicate that some probiotic bacteria could better compete with yoghurt cultures in a soy-based substrate. Soy has been examined as a substrate for the Lactobacillus species such as L. Casei, L. helveticus, L. fermenti, L. fermentum and L. reuteri (Garro et al., 1999; Murti et al., 1993b;( Chumchuere and Robinson, 1999; Garro et al., 2004; Tzortzis et al., 2004).
The problems of which is beany flavor can be improved by lactic fermentation, so production of fermented soymilks such as soy yoghurt is important (Nsofor et al., 1992; AbdEl Khair, , 2009).
Due to continuous increase in the population and inadequate supply of animal milk protein leading to malnutrition in Nigeria many research works have been geared finding alternative protein source from legume.
Soymilk can serve as a very good alternative to the expensive cow milk as it contains all the essential amino acid even though some are in a low concentration
It is well known from experiment that diets containing containing casein or other animal protein could induce elevation of plasma total LDL cholesterol concentration but this can be prevented by vegetable protein such as soy protein (caroll and kurowska 1998)
1.2 HISTORY OF THE YOGURT MAKING
Milk fermentation is one of the oldest methods practiced by the human beings to preserve milk with an extended shelf life. The exact origination of milk fermentation is not clear; however, it seems that it is dated back to the dawn of the civilization.
It has been reported that the early civilizations such as the Samarians, Babylonians, Pharoes and Indians were well advanced in agricultural and animal husbandry practice. This can be supported by the findings of Copley et al., 2003 in which the dairy fat residues were found in pottery fragments from Neolithic Bronze-age and Iron-age settlements, which suggests that the practice of dairying had existed in Britain approximately 6500 years ago.
However, it is questionable that the milk fermentation was practiced during this period. Therefore, the origin of the fermented milk products including yogurt remains unsolved. It has been reported that the Anatolian goatherds conserve their milk by thickening as they used to dry it in the sun and transport in animal stomachs. It is generally accepted among the historians that the fermentation of milk is discovered accidentally by the Neolithic people of Central Asia when they stored milk in primitive methods such as in sheep-skin bags in warm climates.
With reference to yogurt, it can be suggested that it has been evolved in Turkey as the term “yoghurt” has been derived from a Turkish verb, “jugurt” that means “to be curdled or coagulated”.(Belleville, 2002)
The earliest writings about yogurt can be found from those of Pliny who lived in the first century A. D. and wrote about ancient barbarous nations that knew how to thicken the milk into a substance with an agreeable acidity. According to the literature,(Douglass et al.,2006) the founder of the Mongol empire, Genghis Khan and his armies lived on yogurt and spreading of this news among the people had made the yogurt consumption to spread throughout the East.
Moreover, according to the Persian tradition, Abraham owed his fecundity and longevity to the regular ingestion of yogurt, and the emperor Francis I of France was said to be cured of severe diarrhea by consuming yogurt made of goat milk leading to introduce the health benefits of yogurt into the western world in 1542(Howell and Caldwell, 1978)
The first industrialized production of yogurt took place in 1919, in Barcelona, Spain at a company named Danone. Yogurt was firstly introduced to the USA in the early 20th century in the form of tablets especially designed for those with digestive intolerance. However, it became popular in the North America when Danone, a small-scale yogurt factory started manufacture of yogurt in New York in 1940(Fennel, 1966)
Even though, yogurt has been evolved for centuries, it was subjected to a significant and dynamic evolution process in the 20th Century to originate a vast array of products. For instance, fruit yogurts, yogurts with fruit on bottom and blended yogurts were introduced in 1937, 1947 and 1963 respectively. It seems that the evolution process of yogurt has taken place in different regions of the world once it had been originated in the Central Asia. This might be the reason of having different types of yogurts and yogurt-like products in different names.
1.3 STATEMENT OF THE PROBLEM
Animal protein (milk) are more expensive in Nigeria and not easily affordable by all for yogurt production necessitates the need to seek for ways of producing yogurts from plant protein (soy beans).Combination animal protein provides more complete amino acid , hence better nutrition .
Improving product nutrient value by fruit addition and increased new product variety. Reduce postharvest losses of soybean due to underutilization in Nigeria.
Therefore , many researches have been carried out on yogurt but none have being established on using pineapple as sweetener, aimed at achieving the aforementioned solution.
1.4 AIM/OBJECTIVES
To produce yogurt from plant protein (soymilk)
To evaluate the sensory attributes of yogurt made from soymilk and cow milk using pineapple as sweetener.
To determine the proximate composition attributes of the yogurt made from soymilk and cow milk using pineapple as sweetener.
To check if there is a significant difference in the texture, appearance, colour, taste and general acceptability of the yogurt produce using ANOVA
To check the proximate contents of the samples produced
THE EFFECT OF TEMPERATURE ON GLUTATHIONE PEROXIDASE FROM FISH
CHAPTER ONE
INTRODUCTION
- INTRODUCTION
1.1.1 FISH: SOURCE OF FOOD FOR MAN.
Food is one of the basic needs of man (Morey, 1940; Pierce, 2010). Since agriculture produces the food that provides the calories and micronutrients essential for a healthy and productive life, it is interlinked in many important ways to human nutrition and health (Michael, 2011). These nutrients include carbohydrates, proteins, fats and oil, minerals, vitamins and water. Of these nutrients, it is the proteins that supply the body with amino acids necessary for growth and repair of damaged tissues. The sources of protein include plants and animals. Animal sources are preferred because of the presence of essential amino acids and higher digestibility. However, the major disadvantage is higher cost. The animal sources include fish, poultry, dairy, pork, snail, and rabbit.
Fish accounts for one fifth of world total supply of animal protein (FAO, 1991; Olagunjuet al., 2007). It is an important source of protein to a large number of Nigerians. It provides 40% of the dietary intake of animal protein of the average Nigerian (FDF, 1997; Sogbesanet al., 2006). According to Adekoya and Miller (2004), fish and fish products constitute more than 60% of the total protein intake in adults especially in rural areas.
Amiengheme (2005) enumerated the importance of fish in Human Nutrition as follows:
- Fish food has a nutrient profile superior to all terrestrial meats (beef, pork and chicken) being an excellent source of high quality animal protein and highly digestible energy;
- Fish is a good source of sulphur and essential amino acids such as lysine, leucine, valine and arginine. It is therefore suitable for supplementing diets of high carbohydrate contents;
- It is an extremely rich source of polysaturated (Omega III) fatty acids, which are important in lowering blood cholesterol level and high blood pressure, reducing the risk of sudden death from heart attacks and reduces rheumatoid arthritis, lowering the risk of age- related muscular degeneration and vision impairment; decreasing the risk of bowel cancer and reducing insulin resistance in skeletal muscles.
- Fish is also a good source of thiamine fat soluble vitamins (A, D and E), water soluble vitamins (B complex) and minerals (calcium, phosphorus, iron, iodine and selenium);
In Nigeria, fisheries occupy a unique position in the agricultural sector of the economy (Kudi et al., 2008). Its contribution to Gross Domestic Product (GDP) rose from 76.76 billion in 2001 to N162.61 billion in 2005 (CBN Report, 2005). Nigerians are large consumers of fish, with an annual average demand estimate at 1.4 million metric tonnes, (Kudiet al., 2008). Domestic fish production of about 0.5 million metric tonnes is supplied by artisan fishermen (85%), and fish farmers (15%) (Adekoya and Miller, 2004; Emokaro, 2010; BusinessDay, 2011). However a demand and supply gap of at least 0.7 million metric tonnes exists nationally with import making up the short fall at a cost of 400 billion United States dollars per year. According to FAO (2007), this figure (0.7 million metric tonnes) makes Nigeria the largest importer of fish in the developing world.
To take advantage of the large market created by this deficit, the immediate past and current Governments has promoted a backward integration plan for increased fish farming and production in Nigeria, Nigerians are also complementing Government efforts by increasing their participation in aquaculture, with many fish farmers focusing on African flathead catfish, Clariasgariepinus as they have been shown to have a potential market value of two to three times that of other cultivable species like Tilapia and Heterobranchus (FAO, 2000; Fafioye and Oluajo, 2005; Emokaroet al., 2010; Businessday, 2011).
A survey by Addo (2005) revealed that Nigerian children below the age of 18 years, who make up about 47% of our total population are still victims of stunting, wasting and under-weight, so with the increased establishment of more aquaculture in Nigeria, it is possible to reverse this trend of malnourishment among Nigerians in this age bracket.
- THE AFRICAN CATFISH
The African catfish – Clariasgariepinus, an omnivorous freshwater fish, is a popular delicacy in Nigeria. It is a prominent culture species because of its fast growth rate and resistance to diseases and stress factors like over-stocking and poor water quality (Olojoet al., 2005). It is distributed mainly in fresh waters of Africa hence the name African catfish, although it is also seen in Asia. It is named ‘catfish’ because they possess prominent barbels which resemble cat’s whiskers. It has a slender body, flat bony head and broad terminal mouth with four barbels. The pectoral fins have spines. Its dendritic organ is an accessory breathing organ and it is a modification of the gill arches (Ahmed et al., 2008). The adult of about 1.5m weighs up to 29kg (Teugel, 1986).
Habits: They are nocturnal omnivorous animals feeding on living as well as dead organic matter. They are capable of swallowing large prey because of large mouth (Teugel, 1986).They can crawl on the ground to escape drying pools but can also survive in shallow mud for long periods of time between rainy and dry seasons. They can produce croaking sound. They spawn mostly at night in the shallow areas of rivers, lakes and streams. Development of the larva is rapid as they are able to hatch 24 hours and swim within 48 – 72 hours after fertilization.
1.3 ENZYMES AND THE GENERAL EFFECTS OF TEMPERATURE
The living cell is the site of tremendous biochemical activity. Catalysis, through enzymes, makes possible biochemical reactions, which are necessary for all life processes. Enzymes are responsible for bringing about almost all of the chemical reactions in living organisms. Without enzymes, these reactions take place at a rate far too slow for the pace of metabolism.
Like most chemical reactions, the rate of an enzyme-catalyzed reaction is affected by temperature. Variations in reaction temperature as small as 1 or 2 degrees may introduce changes of 10 to 20% in the results. In the case of enzymatic reactions, this is complicated by the fact that many enzymes are adversely affected by high temperatures.
The enzyme Glutathione Peroxidase which catalyses the reduction of hydrogen peroxides is the main focus of this study. Understanding the effects of temperature on the activity of this enzyme in the catfish, could help in determining the best storage temperature for this fish such that harvested catfish can be kept fresh for a longwhile for consumption after
1.4. STATEMENT OF PROBLEM
Despite the clear increase in interest for aquaculture and fish consumption in Nigeria, a major problem of storage exists. This is clearly observed in the lack of fresh iced or well dried catfish options in the market. It is also manifest in the ‘point and kill’ marketing phenomenon, which has invariably restricted catfish farming to retail/subsistence levels intended for consumption by members of the immediate locale were the farm is sited, and discourages large scale catfish farming intended for consumers across state and even national borders.
1.5. RESARCH OBJECTIVES
The aim of this study is to isolate the enzyme Glutathione Peroxidase (GPx) from the liver of the African catfish (Clariasgariepinus) and carryout investigations in vitro to determine
- The optimum temperature range wherein the enzyme Glutathione peroxidase best catalyses the reduction of hydroperoxides and other free radicals, thereby protecting the tissues of the Flat Head Catfish from oxidative stress and damage.
- The temperature range wherein rate of the catalytic reduction activity of the enzyme Glutathione peroxidase is lowest.
1.6. SCOPE OF STUDY
This study will be carried on only the African Catfish (Clariasgariepinus), as survey has shown that it is the by far the catfish variety most consumed by inahbitants of Eastern Nigeria.
1.7. SIGNIFICANCE OF THE STUDY
The importances of this study are as follows:
- Results obtained at the end will accurately advise the best storage temperature for the Flat Head Catfish that will ensure optimum protection of its muscles and other edible tissues from oxidative stress and damage,
- Thereby increasing its shelf life,
- Encourage farmers to engage in larger cultivation of this popular species of fish for wider consumption,
- This in turn reduces production and processing costs; and wastage and loss due to deterioration and decay,
- Ensure that fish which is an important but currently expensive source of protein is made cheaper and consumed more frequently by citizens of all income and age brackets;
- And finally help reduce the malnutrition problem earlier mentioned in the last.
EFFECT OF MOMORDICA CHARANTIA LEAF FRACTIONS ON SERUM ELECTROLYTES AND RENAL FUNCTION INDICES IN ALLOXAN-INDUCED DIABETIC ALBINO RATS
CHAPTER ONE
INTRODUCTION
1.1Background of study
Diabetes affects one hundred and thirty five million people in one year worldwide (King et.al., 1999) and this figure is projected to rise to three hundred million in 2025 ((King et. al.,1998). It is obvious that diabetes, a chronic non communicable disease, continues to have a tremendous impact on society in terms of the quality of life and straining health care resources. The costs incurred in managing or preventing it are enormous, both in Kenya and throughout the world. The disease causes substantial morbidity, mortality and long-term complications and remains a risk factor for cardiovascular disease. In Africa, this disease continues to impact on the poverty levels of the people.
Diabetes mellitus is a systemic metabolic disorder characterized by elevated blood glucose due to absolute oxidative stress may cause tissue to be more susceptible to oxidative damage and progression of disease in renal glomerolus (Brownlee, 2001; Yao et al., 2009).Histopathological evaluations on the diabetic kidney show expansion of mesangial matrix and uniform thickening of basement membranes in glomerulus and tubules (Ziyadeh and Wolf, 2008).
Since ancient times, plants have been a worthy source of medicine, which not only control hyperglycemia at low dosages but can also be taken for longer periods in contrast to synthetic hyperglycemic drugs (Grover et al., 2002). One of these plants is Momordica charantia (MC), also known as karalla, or bitter melon, which belongs to the cucurbitaceafamily, grows in tropical areas, including parts of the Amazon, east Africa, Asia, and the Caribbean, and is cultivated throughout South America as a food and medicine (Grover and Yadav, 2004).
The Momordica charantia (bitter melon) is a widely used plant in the traditional medicine for the treatment of diabetes mellitus (DM). It has been shown that Momordica charantia (Mc) has hypoglycemic effects on animals and humans, however, we don´t know if this effect is present in a chronic time and if the plant extract (stem and leaves) participates in the antihyperglycemic effect.
The Momordica charantia (MC) contains anti-hyperglycemic chemicals include glycosides, saponins, alkaloids, fixed oils, triterpenes, proteins and steroids (Murakami et al., 2001; Erden et al., 2010). These chemicals are concentrated in fruits of the Momordica charantia (MC), therefore fruit of the MC has shown more pronounced anti-hyperglycemic activity (Grover and Yadav, 2004). Presence of antioxidants in the fruits and vegetables such as vitamin C, E, carotenoids, lycopenes and flavonoids are also important in prevent free radical injury (Semiz and Sen, 2007). Total flavonoid and phenol contents ofMomordica charantia (MC) extract were analyzed and revealed that MC extract possess potent diphenylpicrylhydrazyl (DPPH) radical scavenging activity (Wu and Ng, 2008). Several studies have reported the anti-diabetic effects of MC on renal functional and histological changes in alloxan albino rats but only limited data is available on the anti-diabetic effects of MC on renal functional and histological changes in rats. 1.1. Aim
The aim of this study is to investigate the effect of Momordica charantia leaf fractions on some serum electrolytes and renal biomarkers in alloxan-induced diabetic rats.
1.1.0 Specific objectives
i. To obtain fractions of Mormodica charantia leaf modified multi solvent serial extraction
ii. To evaluate the effect of Mormodica charantialeaf fractions on serum electrolytes and other renal function indices in alloxan induced diabetic albino rats.
EFFECTS OF AQUEOUS EXTRACT OF BITTER KOLA (GARCINIA KOLA) ON THE PREGNANCY OUTCOME AND EARLY POSTNATAL DEVELOPMENT OF THE OFFSPRINGS OF DIABETIC PREGNANT RATS
CHAPTER ONE
INTRODUCTION
1.1 INTRODUCTION
Compelling evidence suggests that exposure to an adverse fetal environment may enhance susceptibility to a number of chronic diseases in the future life of the offspring(Buzinaro et al, 2008; Simeoni and Barker, 2009).
Diabetes mellitus is a condition that occur during pregnancy that can substantially influence the development of the offspring in utero and postnatally. Diabetes mellitus is now a pandemic, affecting about 10million Nigerians (Ogbera et al, 2005) and about 350 million people worldwide (Ezzati et al, 2011) among who are pregnant women. It is well documented that the combined stress of diabetes mellitus and pregnancy creates a metabolic environment that is often life threatening to both the mother and the fetus (Freinkel, 1980; Metzger, 1991).
Hence, pregnancy among women that have pre-existing diabetes or gestational diabetes is associated with increased rate of adverse outcome for both mother and fetus (Kingsley, 2007; Shefali et al., 2006).
This is primarily due to altered maternal intrauterine environment, creating a situation in which the fetus is exposed to abnormal metabolic substrate (glucose) levels (Van Assche et al, 1991). There is an increased placental transfer of glucose from mother to fetus because of increased availability at the maternal site (Thomas et al, 1990). The compromised metabolic state of the fetus subsequently precipitates a variety of complications associated with ‘‘fuel-mediated teratogenesis’’ (e.g., hyperglycemia, hyperinsulinemia and macrosomia) (Freinkel, 1980; Metzger, 1991). One particularly devastating effect of diabetic pregnancy is that these conditions affect the fetus not only in utero, but also extend throughout the life of the offspring (Padilha et al, 2007; George et al, 2010). Additionally, maternal hyperglycemia stimulates abnormal fetal growth (Aberg et al, 2001) due to the greater availability of glucose in the blood flow (Maayan-Metzger et al, 2009), and this high weight fetus carries a high risk for 17 developing insulin resistance, glucose intolerance, obesity, and type 2 diabetes mellitus in childhood, adolescence and adulthood (Buzinaroet al, 2008; Simeoni and Barker, 2009)
The chances of reducing this poor outcome of pregnancy among diabetics are intricately related with the level of glycemic control (Shefali et al., 2006).
The fact that the economic cost of managing diabetes mellitus is high confers a very important role to medicinal plants in the management of diabetes mellitus especially in developing countries where resources are meager.
Consequently, a number of plants indigenous to Nigeria have been studied, and found to have hypoglycemic effects. These effects were traced to phytochemicals like alkanoids called active principles that can be extracted from plants(Ojewale, 2006; Osadebe et al., 2004). One of such anti-diabetic plants is Garcinia kola, commonly known as Bitter kola. It is an evergreen tree, indigenous to sub-Saharan Africa and belongs to a family of tropical plants called Guttifera (Ofusori et al., 2008). The seed is a masticatory, used for traditional hospitality in cultural and social ceremonies. Every part of the plant has shown to be of medicinal importance and has a wide range of medicinal effects, hence the name ‘wonder plant’ as it is commonly called.
Among the litany of its medicinal effects are; antidiabetic effects (Iwu et al, 1990), weight reducing effets (Koshy et al., 2001), leptin like action (Hayamizu et al., 2003), antihepatotoxic effects (Akintonwa and Essien, 1990), antioxidative stress and anti DNA 18 damage (Farombi, et al., 2004), detoxification of the toxic effects of other chemicals (Esimoneet al., 2002; Nwokocha et al, 2011), etc.
1.2 JUSTIFICATION FOR THE STUDY
Diabetes mellitus is fast becoming the most common type of disease in school children (Pontiroli, 2004). This may be as a result of the reported more than doubling in therate of diabetes among expectant mothers between 2002 and 2008 (Lawrence et al., 2008).
The economic cost of managing diabetes is high. As a result, in the developing countries where resources are meager, there is a shift from contemporary to orthodox medicine since medicinal plants are relatively easier to find and less expensive alternative.
Coincidentally, some of the commonly consumed plant materials in Nigeria have been found to poses anti-diabetic properties. One of such anti-diabetic plantmaterials Bitter kola seed is used as a stimulant. It is also taken by pregnant women to stop nausea.
Although Bitter kolaseed is a known anti-diabetic, there is paucity of data on;
(1) Its effects on pregnancies complicated by diabetes mellitus
(2) Early postnatal development of the offspring’s of such pregnancies.
Hence, the present study was therefore designed to investigate these.
1.3 AIM
The aim of this study is to determine the effect of consumption of Garcinia kola extract by diabetic pregnant rats on the pregnancy outcome and early postnatal development of their offspring.
1.4 OBJECTIVES
To determine the effect of consumption of Bitter kolaextract by diabetic pregnant rats on;
i. The litter size.
ii. The early postnatal growth of the offsprings (from birth to weaning).
iii. The glucose profile of the offsprings at weanin
1.5 OPERATIONAL DEFINITION OF TERMS
Alloxan diabetes: A type of diabetes induced in rodens by the injection of alloxan hydrate. 20 Pregnancy weight gain:The weight gain during pregnancy.
Programming: The process whereby a stimulus or stress at a critical period of development of the rats results in a lasting or lifelong effect.
Teratogenic: Capable of causing developmental abnormalities in the fetus
Diabetic pregnancy:Pregnancy complicated by diabetes mellitus irrespective of the type.
Early Postnatal development: This is the developmental processes that occur in the offsprings from the time of birth to the weaning day.
Gestation period: The period (in days) between the time when spermatozoa were first seen in vaginal smear and the time of delivery.
Glucose drain: The transfer of glucose from the maternal to fetal blood.
In-utero: Events occurring inside the uterus.
Litter Size: The number of offspring delivered by a pregnant rat.
Litter weight: The weight of the offsprings at birth (g).
Perinatal: Events that occur around the time of birth.
Pregestational (Pre-existing) diabetes: Diabetes existing before pregnancy, irrespective of the type.
Pregnancy weight gain: The weight gain during pregnancy.
Programming: The process whereby a stimulus or stress at a critical period of development of the rats results in a lasting or lifelong effect.
Teratogenic: Capable of causing developmental abnormalities in the fetus..
HYPOLIPIDEMIC AND ANTIOXIDANT CAPACITY OF METHANOL LEAF EXTRACT OF KIGELIA AFRICANA IN ALLOXAN-INDUCED DIABETIC RATS
CHAPTER ONE
INTRODUCTION
1.1 Background of study
Diabetes mellitus is a metabolic disorder resulting from a defect of insulin secretion, which is insulin action or both. Insulin deficiency in turn leads to chronic hyperglycaemia with disturbances of carbohydrate fat and protein metabolism (Kumar et al., 2011).
Globally the estimated incidence of diabetes and project for year 2030, as given by international diabetes federation is 350million (Ananda et al., 2012). Currently available pharmotherapies for the treatment of diabetes mellitus include oral hypoglycaemic agent and insulin. However these current drugs do not restore normal glucose homeostasis and they are free from side effects (Bandawane et al., 2011).
In view of the adverse effect associated with the synthetic drugs and as plants are safer, cheaper, and as much effective. Conventional and anti-diabetic plants can be explored (Kumar et al., 2010). Over 400 traditional plants have been reported for the treatment of diabetes (Ramachandran et al., 2011).
Furthermore after world Health Organisation recommended investigation of hypoglycaemic agents from medicinal plants has become more important (Kumar et al., 2010). Also diabetes has been treated orally with several medicinal plants or their extract based on folklore medicine since ancient times.
Kigelia africana (Lam) Benth (Family: Bignoniaceae) is widely distributed in south central and West Africa. It is known as the cucumber or sausage tree because of its huge fruits (average 0.6cm in length and 44kg in weight) which hang from fibrous stalks. It is also known as balm Khene in Hindi and it is distributed all over India but found in abundance in West Bengal. It is found mostly in water areas and spreads abundantly across wet savannah and riverine areas (Sofowaora et al., 1980).
Experimentally, the plant has shown antibacterial, antifungal, antineoplastic, analgesic, anti-inflammatory and antioxidant properties (Saini et al., 2009). The roots, the wood and leaves have been found to contain kigelinone, vernolic acids, kigelin, iridoids, luteolin and 6-hydroxyluteolin (Picerno et al., 2005). Crude extract of herbs and species and other materials rich in phenolic are of increasing interest in the food industry because they retard oxidative degradation of lipids and thereby improving the quality and nutritional value of food (Frankel, 1995).
The baked fruits of Kigelia africana are used for fermentation of beer. It also has internal application including treatment of dysentery, ringworm, tapeworm, malaria, diabetes, pneumonia, haemorrhage and tooth care (Gills, 1992).
In West Africa, the roots and unripe fruit are used as vermifuge and as treatment for haemorrhoids and rheumatism. The bark is traditionally used as remedy for syphilis and gonorrhoea. The fruits and bark ground and boiled in water are taken orally or used as an enema in treating children’s stomach ailment usually tapeworm (Walt et al., 1962)
1.2 Statement of the problem
As impressive improvement has occurred in global health status in the past century which has become a cause for celebration. Therefore, public health professionals can feel proud of their contribution to these achievements even as they appreciate the complexity of the underlying driving force, many of which lie outside traditional public health work. But this satisfaction must be tempered by emerging concerns (Sen and Bonita, 2000) against the recent evidence suggesting that based current trends many low income countries are unlikely to achieve desired health target by 2015 due to devastating disease and overwhelming failing health system (Travis et al., 2004).
The literature review survey revealed that there is no experimental evidence of antidiabetic and hypolipidemic effect of the plant. Therefore the present work was undertaken to explore the antidiabetic and hypolipidemic potential of Kigelia africana methanol leaf extract of the plant in alloxan induced diabetic rats.
1.3. Aim of the study
The research is aimed at investigating the hypolipidemic and antioxidant capacity of methanol leaf extract of Kigelia africana in alloxan induced diabetic rats.
1.4 Objective of the study
Specifically the study sort to:
- Determine the effects of Kigelia africana methanol leaf extract on antioxidant enzyme.
- Determine the effects of Kigelia africana methanol leaf extract on lipid profile of diabetic rats.
- Determine the effects of Kigelia africana methanol leaf extract on oxidative parameters of alloxan-induced diabetic rats.
INVESTIGATION OF THE PANCREATIC EFFECT OF BUCCHOLZIA CORICACEA FORMULATED DIET IN SUCROSE FED PREGNANT RATS AND THEIR OFFSPRING
CHAPTER ONE
- INTRODUCTION
- BACKGROUND OF THE STUDY
Herbal medicine is the oldest form of health care known to mankind. The use of medicinal plant in the treatment of diseases has been in practice since ancient time in different parts of the world especially in Africa. Plants have always been the most vital source of drugs mainly because most plants are autotrophs, and are able to synthesize a large variety of basic biochemical and organic substances such as carbohydrates, protein, terpenes, steroids, alkaloid and glycosides (N’guessanetal.,2009)
The plant kingdom provides a tremendous reservoir of various chemical substances with potential therapeutic properties. Generally, plants which produce constituents having medical values are designated as medicinal plants (Lawrence et al.,2008). In addition, all plants that taste bitter are used as medicine (Barrett, 2009).
Diabetes mellitus is characterize by insufficient blood levels of the hormone insulin. If the blood concentration of insulin is too low, muscle and liver cells do not absorb glucose from the blood which in turn leads to increase levels of blood glucose (hyperglycemia), impaired metabolism of fats and proteins, ketosis and possible diabetic coma. (willam, et.al 2009)During the past 12 years, the world health organization has been collecting information on the prevalence of diabetes mellitus in adult communities worldwide. Within the age range of 30-64 years, diabetes was found to be absent or rare in some traditional communities in Melanesia, East Africa and South America. In communities of Europeanorigin, the prevalence of diabetes were in the range of 3-10% but migrant indian, Chinese and Hispanic American groups were at higher risk (15-20%). (Adetokunbo, et.al.,2003)
A 2008 study completed in U.S. found the number of America women entering pregnancy with pre-exisiting diabetes is increasing. In fact, the rate of diabetes in expectant mothers has more than doubled in the past six years. (Lawrence,et.al.,2008). This is particularly problematic as diabetes raises the risk of complications during pregnancy, as well as increasing the potential for the children of diabetic mothers to become diabetic in the future
- STATEMENT OF THE PROBLEM
Malnutrition is define as the lack of sufficient nutrients, which are essential for the body’s normal functioning. Over time it affects the bodily organs and results in mild to severe medical problems. One of the malnutrition facts is that the number of hungry people is more in the developing countries. If a pregnant woman is malnourished, it is understandable that the baby in the mother’s womb is not receiving enough nutrient.
Pregnant women who have been through malnutrition, deliver babies with low birth weight. Such children are prone to retarded growth, less coordination, poor vision, learning difficulty, and many other diseases. Anemia is one of the malnutrition dises that affects several pregnant women worldwide.
- AIM OF THE STUDY
To investigate the effect of high sucrose fed pregnant rats in their offsprings and know their pancreatic effect. And also the effect of Buccholozia Coriacea
1.4 OBJECTIVES OF THE STUDY
Induce diabetes by administering sucrose
Administering plants for lowering blood sugar level
Measure the blood and leptin level
OXIDATIVE AND BIOCHEMICAL PARAMETER ASSESSMENT FOR ALLOXAN INDUCED DIABETIC RATS TREATED WITH METHANOL LEAF EXTRACTS OF OCIMUM GRATISSIMUM
CHAPTER ONE
INTRODUCTION
- BACKGROUND OF STUDY
The use of Ocimum gratissimum leaf extract to treat Alloxan induced diabetic rat and its oxidative and biochemical parameter assessments.
In Nigeria, especially in the southern part there is a consumption of plants extracts people consume. They also consume a lot of vegetable in their native diet and some of these plants are believed to cure some aliments. Ocimum gratissimum is one of the many found to lower glycaemia in Type-1 diabetes.
Many components of food such as secondary plants metabolites have been seen to alter biological processes which may reduce the risk of some chronic diseases in human such as diabetes.
Diabetes mellitus has its complications especially diabetes conditions among infected people, Ocimum gratissimum uses against various aliments has gained widespread acceptance in developing as well as developed nations (kolewale et al,. 2011). Diabetes mellitus is an important chronic metabolic disorder of public health concern, it occurs either as a result of pancreatic defects in insulin secretion or the failure of the effectively utilize secreted insulin or both. Hyperglycemia is a common consequence of uncontrolled diabetes, which may over time lead to serious damage to vascular tissue, heart, eye nerve and kidney.
Plants with known and suspected therapeutic potencies have been longed used in the alternative and complementary medicine. Numerous scientific reports exist, describing the relatively low toxicity and effectiveness in selected plants in management of diabetes.
Numerous scientific report exists, descending the relatively low toxicity and effectiveness of selected plants in the management of diabetes. It is unfortunate that a large number of those studies were conducted using either normoglycermeia or animal models.
Inducing of diabetes using either >100mg/kg body weight of Alloxan into these animal models, with recent review showing that Alloxan was one of the most frequently used diabetogenic agent globally.
High dose of the chemical are cytotoxic to the pancreatic beta cells giving rise to insulin deficiency.
- STATEMENT OF THE PROBLEM
The statement of the problem in this research work is that the efficacy of Ocimum gratissimum leaf extract through assaying of biochemical parameters of alloxan-induced diabetic rats.
VISION: To determine the effect Ocimum gratissimum methanol leaf extract has alloxan-induced diabetic rats through assaying of biochemical parameters.
ISSUE STATEMENT: Diabetes mellitus has become one of the prevailing health cases people face today, with no or little knowledge on how it can be controlled and managed, it could worsen beyond control and as far as oxidative damage is concerned, it would multiple the generation of free radicals.
METHODS: Apart from modern drugs that can be used to control and manage diabetes, the use of Ocimum gratissimum menthol leaf extract was employed in other to determine the effect on alloxan-induced diabetic rats through assaying of biochemical parameter.
1.3 AIMS AND OBJECTIVE
1.3.1 AIM OF THE STUDY
The aim of this study is to determine the effects of the methanol leaf extract of Ocimum gratissimum on the oxidative and biochemical parameter of alloxan –induced diabetic rats.
OBJECTIVES OF THE STUDY
Specifically, the study sort to:
- Determine the effects of the extract on MDA level
- Determine the effects of the extract on lipid profile
- Determine the effects of the extract on antioxidant enzyme activity.
THE EFFECT OF BUCCHOLZIA CORIACEA ON RENAL FUNCTION INDICES AND OXIDATIVE STRESS IN SUCROSE FED PREGNANT RATS AND THEIR OFFSPRINGS
CHAPTER ONE
INTRODUCTION
1.1 Background of Study
Malnutrition during pregnancy creates health risks for both the pregnant woman her developing fetus. Not eating enough or eating too much can create health risks, for both the pregnant woman and her developing fetus over nutrition during pregnancy is common in developed countries.
Many woman experience increased appetite during pregnancy. This occurs partly because their hormonal balance is altered, but also because the growing fetus removes food by products from their blood. It is also possible for woman to consume too much during pregnancy and gain excessive weight.
Woman who over consume during pregnancy increase their risk of obesity i.e. woman who gain excessive weight during pregnancy often fail to lose weight after childbirth and risk becoming overweight or obese, pre-exclaims psiai.e a condition which occurs in late pregnancy and s characterized by high levels of protein in the urine, hypertension and excessive fluid in tissues, gestational diabetes. Macros Omnia (fetus over- growth) can occur because a pregnant woman over consumers for example, excessive transfer of glucose and other nutrients can occur in pregnant woman with diabetes. As the nutrition a fetus receives in the womb pregnancies the metabolic system to function later in life over nourished fetuses have an increased risk of obesity and associated metabolic condition 5 such as type 2 diabetes later in life.
There is also evidence of an increased risk of the infant experiencing polycythemias the abnormally high red blood cell concentration seizures.
Eating too much sugar during pregnancy could affect your child’s intelligence and memory (American Journal of preventative medicine) consuming too much sugar acids gain weight. This added to me naturally increasing pregnancy weight, can lead to obesity, which can complicate the delivery. It also leads to containing sugars can lead to the fatty liver syndrome. It can also affect the fetal metabolism in life ( B.Sc Pharm MD and Erick Yoshida, MD MHSc FRCPC). It increases the risk of preeclampsia, high sugar, high sugar intake influences the risk of preeclampsia in pregnant women (EUR JCLIN NUTR. 2012) Aug. 66(8) 920-5 doi:10.103/ESCN 2012.61. EPUB 2012 Jun 20).
Wonderful kola also known as (buchholzia seeds is quickly gaining popularity as it seems to be a cure to almost all ailments. It’s look stands out from the normal kola as it look like a root and not a fruit like the others.
This plant grows in different states including Congo, Nigeria, and others. This plant is known as evergreen, and its seeds are widely used in medicine. However, more frequently it is used in herbal medicine rather than traditional.
1.2 Statement of the problem
The consumption of added sugars (sucrose) over the last 200 years has increased exponentially and parallels the creased prevalence of chronic kidney disease. Data for animals and humans suggest that the consumption of added sugars leads to kidney damage and related metabolic derangements that increase cardiovascular risk. Importantly, the consumption of added sugars has been sound to induce insulin resistance and increase uric acid in humans, both of which increase the conversion of glucose and fructose via the payola pathway. The payola pathway has recently been implicated in the contrition and progression of kidney damage.
To mitigate this serious complications and a negative outcome of kidney disease, the plant Buccholzia coriacea can be used as therapeutic option readily available to cure the damage caused by high sucrose.
1.3 Objective of the Study
The general of the, this study was to evaluate the biochemical/metabolic effect of sucrose on a sucrose fed pregnant rat and their offspring’s. The specific objectives are to
- to induce the rats with sucrose
- Carryout biochemical assays including, serum urea nitrogen and lipid per oxidation, serum creatinine.
1.4 Significant of the study
This study will increase our understanding on the renal effect of buccholzia coriacea on the kidney function indices of a sucrose fed pregnant rats. It is hope that constituent compounds present in buchholziacoriacea would aid further scientific investigation while contributing to the drug discovery and development.
THE RELATIONSHIP BETWEEN ANAEMIA, MALARIA AND ANTIOXIDANT STATUS IN PATIENT VISITING ENUGU STATE UNIVERSITY TEACHING HOSPITAL “ESUTH”
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background of the Study
Malaria is a common and life threatening disease in many tropical and subtropical areas and the term malaria originates from medieval Italian malaran: mala-aria “bad air”; the disease was formerly called ague or marsh fever due to its association with swamps and marshland. The term first appeared in the English literature about 1829. (Breeveld et al., 2012) There are currently over 100 countries and territories where there is high risk of malaria transmission.
Malaria is one of the most important tropical infectious diseases. The annual worldwide incidence is estimated to be 300–500 each year with a mortality of between one and three million people. (Asante et al., 2004).
According to the World Health Organization (WHO), malaria is a significant public health problem in more than 100 countries and causes an estimated 200 million infections each year, with more than 500 thousand deaths annually. Over 90% of these deaths occur in sub-Saharan Africa, where the disease is estimated to kill one child every 30 seconds (WHO, 2011), and Malaria is the 3rd leading cause of death for children under five years worldwide.
Malaria in Nigeria Malaria is a major public health problem in Nigeria where it accounts for more cases and deaths than any other country in the world. Malaria is at risk to 97% of Nigeria’s population. The remaining 3% of the population live in the malaria free highlands. (Kremsner et al., 200). There are an estimated 100 million malaria cases with over 300,000 deaths per year in Nigeria. This compares with 215,000 deaths per year in Nigeria from HIV/AIDS. Malaria contributes to an estimated 11% of maternal mortality. (WHO, 2012).
Malaria accounts for 60% of outpatient visits and 30% of hospitalizations among children under five years of age in Nigeria. Malaria has the greatest prevalence of 27.6 percent, in children age 6 to 59 months in the South East region. (Abubakar et al., 2002).
Malaria is a significant global problem. In 2015, there were 214 million cases of the disease worldwide, killing about 438,000 people. In other areas of the world, malaria causes substantial morbidity, especially in the rural areas of some countries in Asia and South America , and these countries are visited by more than 125 million international travelers every year and malaria is most often cause by travel to and from endemic areas. (Abubakar et al., 2002).
Malaria is a mosquito-borne infectious disease affecting humans and other animals caused by parasitic protozoan’s (a group of single-celled microorganisms) belonging to the Plasmodium types like Plasmodium, falciparum, Plasmodium, vivax, Plasmodium, ovale, Plasmodium, malariae and transmitted to humans/ animals by the bite of infected mosquitoes (WHO, 2014). The malaria parasite, entering the blood after an infective mosquito bite, which will infects/invades the red bloods cells. At the end of that infection cycle, red blood cell ruptures. The process lowers the amount of red blood cells and can cause anaemia which is low haemoglobin levels, frequently leading to anaemia. Plasmodium facliparumcauses the most severe and profound anemia Malaria infection in human by plasmodium species is associated with a reduction with a significant risk of death, this cannot be explained simply by the direct destruction of parasitized red blood cells.
Malaria infection induces the generation of hydroxyl radicals (OH•) in the liver, which most probably is the main reason for the induction of oxidative stress. (Becker K et al., 2004). It was observed that erythrocytes infected withPlasmodium. falciparum produced OH• radicals and H2O2 about twice as much compared to normal erythrocytes. Higher level of these free radicals can lead to oxidative stress. (Becker K et al., 2004).
Oxidative stress, termed as an imbalance between production and elimination of reactive oxygen species (ROS) leading to plural oxidative modifications of basic and regulatory processes, can be caused in different ways. Increased steady-state ROS levels can be promoted by drug metabolism, over expression of ROS-producing enzymes, or ionizing radiation, as well as due to deficiency of antioxidant enzymes as well as by malaria parasite. The consequence of oxidative stress once it is high, it can attack cellular membrane lipids, causing damage to cells and tissues such as the brain, metabolic disorders or inherited disease affecting electron transport chain.
Oxidative stress during malaria is considered useful to the patient in the fight against the intra-erythrocytic parasite (Gilbert D.L. 1981). Studies have been described in which induction of oxidative stress by treatment with pro-oxidants proved to be effective against the infection. On the other hand ROS play a role in the pathology of malaria (Jayshree et al., 1993). Excessive oxidative stress particularly at unprescribed sited (e.g. vascular lining, blood brain barrier) can damage the defense system. This is however, controlled by intra- and extracellular anti-oxidants systems, which may fail during disease. Treatment with anti-oxidants reinforces these systems and protects the patient, especially during the life threatening phase of the disease (Rice-Evans et al., 1992).
In malaria infections, the most probable target of free radical generated by malaria parasite is the red blood cell since it is where the parasite resides. Thus, though malaria parasite infection also damage the red blood cell during its asexual stage multiplication, free radicals produced by the parasite may also contribute. Thus, it may be suggested that anaeamia which is defined by the reduction of haemoglobin in blood below normal may result from the parasite damaging the red blood cell as well as free radical generation Anemia is a condition that occurs when the amount of haemoglobin in a person’s drops below normal .and a decrease in haemoglobin is often associated with a decrease in the number of red blood cells and haematocrit. Haemoglobin is contained with RBCs and it is necessary to transport and delivery of oxygen from the lungs to the rest of the body. Without a sufficient supply of oxygen, many tissues and organs throughout the body can be adversely affected. People with anemia may experience faigue weakness / tiredness, and may lack energy, breathlessness. groups , However, certain people have increase risk of developing anemia Anaemia fairly common condition affecting both men and women of all ages, race and ethnic. These include people with diets poor in irons and vitamins, chronic diseases such as kidney disease and inflammatory bowel disease. Therefore, increased level of anaemia in malaria may be due to the influence of free radical induced oxidative stress.
Antioxidants are molecules that inhibit or quench free radical reactions and delay or inhibit cellular damage (Adam-Vizi, V. (2005). Though the antioxidant defenses are different from species to species, the presence of the antioxidant defense is universal. Antioxidants exists both in enzymatic and non-enzymatic forms in the intracellular and extracellular environment. Enzymatic antioxidants work by breaking down and removing free radicals. The antioxidant enzymes convert dangerous oxidative products to hydrogen peroxide (H2O2) and then to water, in a multi-step process in presence of cofactors such as copper, zinc, manganese, and iron. Non-enzymatic antioxidants work by interrupting free radical chain reactions (Adam-Vizi, V. (2005). The major enzymatic antioxidants are superoxide dismutase, catalyze and glutathione peroxidase. Superoxide dismutase exists as a copper, zinc-enzyme (SOD1) that is found in the cytoplasm or a manganese containing enzyme that is located in mitochondria (SOD2). These enzymes catalyze the one-electron dismutation of superoxide (O2·) to hydrogen peroxide (2O2 + 2H+ → H2O2 + O2). Catalyse is an iron-dependent enzyme that directly decomposes hydrogen peroxide to water (2H2O2 → 2H2O + O2). Furthermore, glutathione peroxidases (GPXs) are a family of enzymes that reduce a variety of organic and inorganic hydroperoxides to the corresponding hydroxyl derivatives in the presence of glutathione (GSH). In this process, GSH is converted to an oxidized disulfide (2GSH + H2O2 → GS-SG + 2H2O).
The other way of categorizing the antioxidants is based on their solubility in the water or lipids. The antioxidants can be categorized as water-soluble antioxidants. The water-soluble antioxidants (e.g. vitamin C) are present in the cellular fluids such as cytosol, or cytoplasmic matrix.
1.2 Statement of the problem
Malaria remains one of the leading causes of morbidity and mortality worldwide and in sub- Saharan Africa. (WHO, 2012). Mortality from malaria is due to complication arising as a result of severe infections usually caused byPlasmoduim Falciparum. Studies on mortality have shown that deaths occur predominantly among young children/ some adult and mortality rates among patients with an illness severe enough to warrant hospitalization are consistently high with case fatality rates varying from 5% to 30% in Nigeria, malaria is hyper endemic and presents a serious health problem in the country. It is also a leading cause of deaths in the country and accounts for over 40% of out-patient attendance with annual reported cases and children less than five years are the most affected. A study conducted by Ministry of Health in 2006 showed that more than 17 million of Nigerian’s over twenty million people are infected with malaria/anaemia every year, with cost of $95 million for treatment. Despite the importance of Plasmoduim Falciparum. As a human pathogen, the patho-phyrsiologic basis of its infection is not well understood. Parasitic infections such as malaria in host organisms often lead to anaemia and oxidative stress condition which is a disturbance in the balance between the production of ROS and antioxidant defenses .generation of free radicals as a result of oxidative stress and other reactive species in vivo leads to extensive damage in parasite bio- molecules such as DNA, lipids and proteins. It has also been shown that the parasites are vulnerable to oxidative stress during their erythrocytic life stages. In erythrocytes, Plasmoduim Falciparum. Encounters enhanced oxidative stress, resulting largely from its digestion of haemoglobin and thus, its redox balance becomes fragile. Superoxide (O2-) is normally produced when oxidized haemoglobin is exposed to the acid environment of the food vacuole, and can therefore be considered as the major source of ROS. Inside the parasite, regardless of its origin, O2– is dismutated by superoxide dismutase (SOD) to H2O2(Makani, J. et al., 2010). Even though studies have been carried out on Plasmoduim Falciparum and ROS, the focus has mainly been on the pathological effect of these radicals.
1.3 Hypothesis
Malaria may promote oxidative stress and anaemia and reduce antioxidants.
1.4 Aim of the study
The study was undertaken to evaluate the relationship between anaemia, malaria infection, and antioxidant status of patients visiting Enugu State University Teaching Hospital (ESUTH) in Enugu Nigeria.
1.5 Objective of the study
- To determine the presence of malaria parasite infection in patients
- Determine haemoglobin concentration and haemoglobin to assess anaemia in infected patients.
- Determine antioxidant indices (SOD, and Vitamin C) among malaria patients.
RELATIONSHIP BETWEEN ANAEMIA, GLUCOSE-6-PHOSPHATE DEHYDROGENASE DEFICIENCY AND OXIDATIVE STRESS AMONG MALARIA PATIENTS VISITING ESUT TEACHING HOSPTIAL ENUGU NIGERIA
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background of study
Malaria is a mosquito-borne infectious disease affecting humans and other animals caused by parasitic protozoans (a group of single-celled microorganisms) belonging to the Plasmodium type (WHO, 2014). According to the World Health Organization (WHO), malaria is a significant public health problem in more than 100 countries and causes an estimated 200 million infections each year, with more than 500 thousand deaths annually. Over 90% of these deaths occur in sub-Saharan Africa, where the disease is estimated to kill one child every 30 seconds (WHO, 2011). In other areas of the world, malaria causes substantial morbidity, especially in the rural areas of some countries in Asia and South America. Malaria causes symptoms that typically include fever, tiredness, vomiting, and headaches. In severe cases it can cause yellow skin, seizures, coma, or death (Caraballo, 2014). Symptoms usually begin ten to fifteen days after being bitten, If not properly treated, people may have recurrences of the disease months later. The disease is most commonly transmitted by an infected female Anopheles mosquito. The mosquito bite introduces the parasites from the mosquito’s saliva into a person’s blood (WHO, 2014). The parasites travel to the liver where they mature and reproduce. Five species of Plasmodium can infect and be spread by humans. (Caraballo, 2014). Most deaths are caused by Plasmodium falciparum.
The role of oxidative stress during malaria infection is still unclear. Some authors suggest a protective role, whereas others claim a relation to the physiopathology of the disease (Sohail et al., 2007). However, recent studies suggest that the generation of reactive oxygen and nitrogen species (ROS and RNS) associated with oxidative stress, plays a crucial role in the development of systemic complications caused by malaria. Malaria infection induces the generation of hydroxyl radicals (OH•) in the liver, which most probably is the main reason for the induction of oxidative stress and apoptosis (Guha et al., 2006). Additionally, Atamna et al. (1993) observed that erythrocytes infected with P. falciparumproduced OH• radicals and H2O2 about twice as much compared to normal erythrocytes. Higher level of this free radicals can lead to oxidative stress.
Oxidative stress, termed as an imbalance between production and elimination of reactive oxygen species (ROS) leading to plural oxidative modifications of basic and regulatory processes, can be caused in different ways. Increased steady-state ROS levels can be promoted by drug metabolism, over-expression of ROS-producing enzymes, or ionizing radiation, as well as due to deficiency of antioxidant enzymes. The consequence of oxidative stress once it is high, it can cause damage to the brain, metabolic disorders affecting electron transport chain. Reactive oxygen species (ROS), generated by endogenous and exogenous sources, cause significant damage to macromolecules, including DNA (Salmonet al., 2004).
Furthermore, Spermatozoa are highly vulnerable to oxidative attack because they lack significant antioxidant protection due to the limited volume and restricted distribution of cytoplasmic space in which to house an appropriate armoury of defensive enzymes. In particular, sperm membrane lipids are susceptible to oxidative stress because they abound in significant amounts of polyunsaturated fatty acids. Susceptibility to oxidative attack is further exacerbated by the fact that these cells actively generate reactive oxygen species (ROS) in order to drive the increase in tyrosine phosphorylation associated with sperm capacitation. However, this positive role for ROS is reversed when spermatozoa are stressed. Under these conditions, they default to an intrinsic apoptotic pathway characterised by mitochondrial ROS generation, loss of mitochondrial membrane potential, caspase activation, phosphatidylserine exposure and oxidative DNA damage. In responding to oxidative stress, spermatozoa only possess the first enzyme in the base excision repair pathway, 8-oxoguanine DNA glycosylase. This enzyme catalyses the formation of abasic sites, thereby destabilising the DNA backbone and generating strand breaks. Because oxidative damage to sperm DNA is associated with both miscarriage and developmental abnormalities in the offspring, strategies for the amelioration of such stress, including the development of effective antioxidant formulations, are becoming increasingly urgent (Aitken et at., 2016).
The process of lipid peroxidation involves a complex chain reaction utilizing the interaction of oxygen-derived species with polyunsaturated fatty acids (e.g. docosahexaenoic acid, linoleic acid and arachidonic acid), resulting in highly reactive electrophilic aldehydes and free radicals (Esterbauer et al., 1991). This process is extremely detrimental to cellular functions as it disrupts membrane integrity, fluidity and function (Esterbauer et al., 1991). Lipid peroxidation is a self-propagating process involving initiation and propagation steps which continue through an ongoing free radical chain reaction until termination occurs. The retina is particularly prone to lipid peroxidation since it is highly enriched in polyunsaturated fatty acids (PUFAs) (Catalase). The predominant PUFA in photoreceptor outer segments is docosahexanoic acid which is the most unsaturated fatty acid in the body. Lifelong accumulation of chronic oxidative damage will lead to dysfunction in retinal cells and increase their susceptibility to exogenous and endogenous insults eventually culminating in loss of visual function and cell death (Esterbauer et al.,1991). Malaria infection has been found to be associated with lipid peroxidation accompanying reduction in antioxidant capacity of the infected patients especially Plasmodium falciparum infection. Instantaneous reduction in antioxidant potency in tandem with increased lipid peroxidation is also observed to be equally accountable for development of oxidative stress in malaria patients (Das and Nanda, 1999; Upadhyay et al., 2011; Egwunyenga et al., 2004). Any infection, including malaria, activates the immune system of body thereby causing release of reactive oxygen species as an antimicrobial action (Kulkarni et al., 2003). In addition to host’s immune system, malaria parasite also stimulates certain cells in production of reactive oxygen species thereby resulting in hemoglobin degradation (Loria et al., 1999; Pradines et al., 2005). One of the major reasons for development of malarial anemia seems to be oxidative stress (Das and Nanda, 1999; Kremsner et al., 2000) while changes in micronutrient metabolism alter disease progression and severity (Singotamu et al., 2006).
Proteins are the largest constituent of the cellular milieu and are frequent targets of oxidative damage (Stadtman, 2004). Protein oxidation can involve direct reaction with amino acids, cleavage of the polypeptide chain, and conversion of the protein to derivatives that are highly sensitive to proteolytic degradation. It has also been established that all of these protein modifications can be mediated by metal-catalyzed oxidation systems. All amino acid residues of proteins are potential targets for oxidation by HO· or by H2O2 in the presence of metal ions. For example, oxidation of tyrosine residues is damaging to the red blood cells, as this amino acid is converted to a 3,4-dihydroxyphenylanine derivative, which itself can undergo redox cycling to generate further ROS (Sugiura and Ichinose, 2011).
Antioxidants are molecules that inhibit or quench free radical reactions and delay or inhibit cellular damage (Young et al., 2001). Though the antioxidant defenses are different from species to species, the presence of the antioxidant defense is universal. Antioxidants exists both in enzymatic and non-enzymatic forms in the intracellular and extracellular environment. . Enzymatic antioxidants work by breaking down and removing free radicals. The antioxidant enzymes convert dangerous oxidative products to hydrogen peroxide (H2O2) and then to water, in a multi-step process in presence of cofactors such as copper, zinc, manganese, and iron. Non-enzymatic antioxidants work by interrupting free radical chain reactions (Young et al.,2001).
The antioxidants can also be categorized according to their size, the small-molecule antioxidants and large-molecule antioxidants. The small-molecule antioxidants neutralize the ROS in a process called radical scavenging and carry them away.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathological disease in humans. This disease is described as a widespread, heritable, X-chromosome linked abnormality (Reclose et al., 2000). It is estimated that it affects approximately 400 million people worldwide (Daloii et al., 2004). This disease is seen most frequently in approximately all of Africa, Asia, and the countries near the mediterranean Sea (Frank, 2005). G6PD enzyme was demonstrated to play an active role in survival of erythrocytes. It is known that in the pentose phosphate pathway of erythrocytes, glucose-6 phosphate dehydrogenase (G6PD) enzyme provides the production of NADPH and Glutathione (GSH). GSH, produced by pentose phosphate pathway can react with H2O2 and reduce it to H2O. This prevents the generation of oxidative stress within red blood cells; oxidative stress can be induced in erythrocytes whose G6PD enzymes are deficient. In this situation, GSH is not produced and H2O2 is not reduced to H2O, leading to oxidative stress and hemolysis.
1.2 Justification / Need for study
The need for this study is to carefully ascertain whether G6pD deficiency has an impact on oxidative stress and malaria infection. To known if G6pD can promote malaria infection in patient. This study is equally designed to give information about the prevalence of G6PD and malaria infection among malaria patient in Enugu metropolis.
1.3 Statement of the problem
So many reasons have been attributed to susceptibility of G-6-PD deficient patients to malaria infection. Recall that the pentose phosphate pathway is essential in producing enough NADPH capable of reducing oxidized glutathione but when there is a default in the production of enough NADPH from PPP as the case may be in G-6-PD deficient patients, leading to a decrease in the level of reduced glutathione which leads to an increased in oxidative stress – a possible risk factor in the development of malaria infection will be complicated.
1.4 Aim of the study
This study was undertaken to evaluate the relationship between oxidative stress among malaria patients visiting Enugu State University Teaching Hospital, Parklane, Enugu Nigeria.
1.5 Objective of the study
1. To screen patients for malaria infection.
2. To assess anaemia in malaria patients.
3. To assess G6PD deficiency in malaria patients.
4. To assess oxidative stress indices (lipid peroxidation and protein oxidation) in malaria patients.
EFFECTS OF SELENIUM TOXICITY AND DEFICIENCY ON HUMANS
CHAPTER ONE
INTRODUCTION
1.1. BACKGROUND INFORMATION
Selenium (Se) is an essential trace element having biological functions of utmost importance for human health. Different from the other (semi) metals, it is incorporated into proteins by a co-translational mechanism as part of the amino acid selenocysteine (SeCys), the 21st amino acid used for protein synthesis in humans, whereas only a few of them have been functionally characterized. Most Se-proteins participate in antioxidant defence and redox state regulation, particularly the families of more specific essential roles, such as iodothyronine deiodinases (DIOs) which are involved in thyroid hormones metabolism, GPx4 which is essential for spermatogenesis, and selenophospathe synthetases 2 (SPS2) participating in Se-protein biosynthesis.
Other Se-proteins may be involved in important biological processes, but their exact mechanism of action is still yet to be fully understood. Despite the scarce knowledge of the precise biochemical functions, a very large number of studies have been carried out in the last two decades showing that insufficient Se levels, and particularly Se-proteins, are associated with several human diseases including cancer, diabetes, cardiovascular and immune system disorders. In most cases, the link lies in the contrast to the oxidative stress that may be booth causing or caused by the disease. In this context, it is important to decipher whether and adequate Se status may contrast the risk factors for health disorders, or Se supplementation may improve the therapy when Se metabolism is altered.
Despite many studies that have suggested a beneficial effect from Se supplementation to general health protection, most of them have remarked that it is limited to general health protection, most of them have remarked that it is limited to the initially inadequate Se status. Conversely, care should be taken when using supplements because excessive Se intake leads to toxic effects, and recent studies have shown that even sub-toxic doses may be negatively impacting, for example by increasing the risk of type 2 diabetes.
EVALUATION OF NUTRITIVE AND NON-NUTRITIVE COMPONENTS AND ANTI-DIABETIC PROPERTIES OF EDIBLE PORTIONS OF Chrysophyllum albidum FRUIT IN STREPTOZOTOCIN-INDUCED DIABETIC MALE ALBINO RATS
CHAPTER ONE
INTRODUCTION
1.1 Background to the Study
Dietary control is vital in the management of diabetes. Reports from FAO (1998); WHO and FAO (2003) have shown that diets with low saturated fat, cholesterol and glycemic index as well as high contents of soluble fiber, vitamins and minerals are effective in the management of diabetes. Low glycemic foods contain sugars that digest and absorbed slowly into the blood and thus control blood sugar levels. The fiber-like substances such as gums and pectin reduced blood postprandial glucose levels (Jenkins et al., 1978; Ou et al., 2001) while diabetic subjects fed xanthan gum have lower fasting, postprandial serum glucose levels and total plasma cholesterol (Osilesi et al., 1985). Jenkins et al. (1978) reported that daily intake of 5–10 g of soluble fiber from different sources reduced serum cholesterol by 5–10%. Fruits serve as one of the best sources of dietary fiber, minerals, Vitamins A, C and E and frequent intake of vegetables and fruits have demonstrated a lowered risk of diabetes, heart disease, hypertension, stroke and cancer (Southon, 2000; Wargovich, 2000). Fruits supply carbohydrates in the form of soluble sugars, cellulose and starch (Nahar et al., 1998) and serve as source of nutrient, appetizer and food supplement in a world faced with problem of food scarcity.
Diabetes mellitus (DM) is a worldwide endemic disease in terms of occurrence, cost of medical care, and general complications (King et al., 1998). The metabolism of protein, carbohydrate and fat are affected in diabetic conditions, resulting in hyperglycemia. DM complication is mainly associated with a high risk of coronary heart disease (Giugliano et al., 1996), atherosclerosis, stroke and peripheral vascular disease. The incidence of DM world wide, is projected to increase from 4% in 1995 to 5.4% by the year 2025 (Mohamed et al., 2006), with the utmost increases set to occur in the developing countries of Africa, Asia and South America (WHO, 2008).
According to WHO (1994) and American Diabetes Association (2008), diabetes mellitus can be classified into insulin-dependent diabetes mellitus, IDDM (type 1 diabetes mellitus) and non- insulin- dependent diabetes mellitus, NIDDM (type 2 diabetes mellitus). Insulin-dependent diabetes mellitus is caused by cellular-mediated autoimmune damage to beta cells of the pancreas, accounts for about 5% to 15% of diabetic cases and occurs mostly in children or adolescents (Ranjan and Ramanujam, 2002). Genetics and environmental factors are implicated in the formation of IDDM. Administration of exogenous insulin is thus required to avert ketosis and preserve life (Lokesh and Amit, 2006). Non- insulin- dependent diabetes mellitus starts as insulin resistance, accounts for 85-95% of cases globally and occurs usually in adults of 40 years and above (WHO Regional Office for the South-East Asia, 2009). It is associated with hyperglycemia and glycosuria. The risk factors increases with age, lack of physical activity, obesity and impaired glucose tolerance.
Insulin resistance occurs when glucose is not properly utilized by the cells leading to high blood glucose in circulation. To maintain blood glucose level, the kidney excretes exess blood glucose through the urine and glucosuria occurs with increased excretion of water and sodium when blood glucose level exceeds the renal threshold (160 – 180 mg/L). The failure to use glucose by the body cells, results to increase appetite (polyphagia) (Robinson et al., 1986). The summary of the symptoms of diabetes is shown in Figure 1.
Figure 1: Overview of the most Significant Symptoms of Diabetes
Source: Cooke and Plotnick (2008)
Insulin resistance is associated with decreased glucose uptake and stimulation of muscle glycogen synthesis (Cline et al., 1999). In addition, alteration of enzymatic activities like increased phosphatase activity and/or seryl phosphorylation of the insulin receptor substrate by glycogen synthesis kinase 3 (GSK- 3), have also been reported in some cases of type 2 diabetes mellitus (Begum et al., 1991; Nadiv et al.,1994; Eldar-Finkelman and Krebs, 1997). Insulin resistance plays an important role in the etiology of many disorders including obesity, NIDDM, glucose intolerance, hypertension and other related disorders. It has been reported that autophosphorylation of insulin receptor kinase and subsequent phosphorylation of its principal substrate, IRS-1, are significantly lowered in insulin-responsive tissues of patient with severe obesity or NIDDM (Nadiv et al., 1992). Increased lipolysis and decreased lipogenesis occurred when there is a fall in circulated insulin leading to fatty acids release from adipose tissues and subsequently oxidized to ketone bodies in the liver. The rapid release of fatty acids into the blood leads to increase level of blood cholesterol and the formation of atherosclerosis (Khan and Ahmad, 1993). In diabetics, there is increase in excreted nitrogen through deamination, which is accompanied by cellular potassium excretion in urine when the muscle protein is broken down to support gluconeogenesis in the liver.
Of the several approaches applied, to lower and control the occurrence of diabetes, drug and diet therapies form the most popular approaches. The most common approach are the drug therapy with four distinct classes of oral hypoglycemic agents (biguanides, sulfonylureas, thiazolidinediones and alpha-glucosidase inhibitors) currently being recommended for use to treat NIDDM. In dietary therapy, dietary modifications with adequate exercise are used to prevent excessive weight gain and obesity (Derek, 2001). Intake of diets with low total and saturated fat, limited protein with replacement by complex carbohydrate and/or mono unsaturated fatty acids are the recommended diets for type 2 diabetes patients. Controlled diets will improve the metabolic control in diabetic subject and lower the risk of diabetes complications (Griver and Henry, 1994).
A large number of plants with hypoglycemic activity have been reported in different animal models. Aloe vera, Acacia arabica, Allium sativum L., Bombax ceiba L., Allium cepa, Brassicajuncea (L.) Cassia auriculata L., Caesalpinia bonducella (L.) and Musa sapientum L. are some of the scientifically validated antidiabetic plants (Modak et al., 2007).
Plant of Study
Chrysophyllum albidum (Linn), commonly called African star apple is a forest tree species of Sapotaceae family (Figure 2). It is widely distributed in Nigeria, Niger Republic and Uganda (Bada, 1997). C. albidum has various ethno-medicinal uses (Dalziel, 1937; Amusa et al., 2003) and across Nigeria, it is locally called ‘‘agbalumo’’ in South Western Nigeria and “udara” in South Eastern Nigeria.
Figure 2: Chrysophyllum albidum Tree
Source: Orwa et al. (2009)
The fleshy pulp of C. albidum fruit is taken as snack, the seeds serve as a source of oil for various uses and the fruit is a good source of ascorbic acid (Adisa, 2000; Adepoju and Adeniji, 2012). C. albidum plants are rich in natural antioxidants and can thus support health by preventing oxidative stress related disease such as diabetics, cancer and coronary heart diseases (Burits and Bucar, 2002). The antioxidants content in vegetables and fruits has been associated with the diminished risk to chronic diseases by scavenging free radicals and prevent cells damage (Halliwell, 1994).
The antimicrobial and phytochemical screening of C. albidum seed cotyledon (Idowu et al., 2003; Okoli and Okere, 2010), leaves (Duyilemi and Lawal, 2009; Okoli and Okere, 2010; Kamba and Hassan 2011), root (Okoli and Okere, 2010), and stem bark (Adewoye et al., 2010; Kamba and Hassan 2011) have been investigated. In addition, the anti-hyperglycemic and hypolipidemic effects of C. albidum seed cotyledon ethanolic extract (Olorunnisola et al., 2008) and leaf ethanolic extract (Adebayo et al., 2010) have been reported. Adebayo et al., 2010, 2011a and 2011b, have reported the antiplatelet, antioxidant and hepatoprotective effects of C. albidum leaf while Onyeka et al. (2012) and Omotosho et al. (2013) reported the antifertility and antioxidant effects of C. albidum root bark and fruit juice.
Nwadinigwe (1982); Edem et al. (1984); Adisa (2000); Ige and Gbadamosi (2007); Ureigho (2010); Christopher and Dosunmu (2011); Oyebade et al. (2011); Adepoju and Adeniji (2012), have independently analyzed the nutritional contents of C. albidum pulp. Similarly, Ige and Gbadamosi (2007) analyzed the nutrient compositions of C. albidum fruit-peel (skin) and fruit juice. Ewansiha et al. (2011), analyzed C. albidum seed shell pericarp for its nutritional compositions while Ajewole and Adeyeye (1990), studied the physico-chemical characteristics and fatty acid composition of the seed. However, information on the nutrient contents of seed shell pericarp, fruit skin (peel) and fruit pulp of C. albidum are scanty in available literature. In addition, there is dearth of information on the efficacy of either of these edible portions of C. albidum fruit as remedy for the management of DM. Therefore, this study was design to investigate the nutrtitive and non-nutritive components and the antidiabetic potentials of the edible portions of C. albidum fruit.
1.2 Statement of the Problem
Diet has a vital role in the causes and control of several obesity-associated chronic diseases, such as diabetes and cardiovascular diseases. Current research has increased on studying individual foods to understand their specific role(s) and the mechanisms of action in the diminished risk to diseases in humans. Diabetes has emerged into a global epidemic, inspite of the recent search in new drugs to manage and prevent the condition; its prevalence continues to soar with increased risks and diagnosis in both adult and children (Ludwig and Ebbeling, 2001). In addition, many synthetic hypoglycemic agents such as biguanides, sulfonylureas, α-glucosidase inhibitors and insulin, commonly used for the treatment of diabetes are expensive and associated with serious side effects (Gupta et al., 2010). Sulfonylureas (e.g., glibenclamide) cause severe hypoglycemia, biguanides (e.g., metformins) are unsafe for patients with kidney problem, while α-glucosidase inhibitors cause dose-related malabsorption, flatulence and abdominal bloating (Codario, 2005). In addition, these hypoglycemic agents are not effective in the control of hyperlipidemia condition, which usually accompanies the incidence of diabetes (Derek, 2001). These associated problems with the synthetic oral anti-diabetic agents in terms of inefficacy, non-safety coupled with the emergence of the disease into a global epidemy have necessitate the search for more efficient alternatives with little or no side effect (Ranjan and Ramanujam, 2002). The plant kingdom, thus become a target for the search to develop indigenous, inexpensive botanical sources by multinational drug and biologically active lead compounds (Evans, 1996).
Since ancient times, medicinal plants with various active principles and properties have been used by laymen and physicians to cure a variety of human diseases such as coronary heart disease, diabetes and cancer (Havsteen, 1984; Middleton et al., 2000). Medicinal plants offer exciting opportunity to develop them into novel therapeutics due to their multiple beneficial effects as manipulating carbohydrate metabolism by various
COMPARATIVE QUANTITATIVE PHYTOCHEMICAL ANALYSIS OF ETHANOL EXTRACT OF Gongronema latifolium AND Allium sativum
CHAPTER ONE
INTRODUCTION
1.1 Background of the study
The tropical rainforest is the most biologically varying ecosystem on earth and it is enriched with enormous natural plant resources with rich dietary and medicinal properties utilized locally in folkloric medicine (Dalziel, 1937). Although modern medicine may be available in developing countries, the use of herbs in treatment of diseases has often gained popularity for historical and cultural reasons (Nwangwu et al., 2009), making traditional medicine an unavoidable global discuss. The use of medicinal plants is one of the primary complementary advances to medicine in many parts of the world (American Society for Testing and Materials, 2001). The presence of wide range of bioactive phytochemicals and secondary metabolites has made plants promising source of modern synthetic drugs for management of several diseases. One of the basic principles of medicinal herbs is that all constituents in the whole plant extract work collectively to achieve therapeutic efficacy (WHO, 1996). The use of plant extracts in treatment and prevention of diseases have shown a comprehensive framework of healing by stimulating the body’s own innate healing capacities in a manner that is safe and effective (Tietz, 1976).
Gongronema latifolium is commonly called Utazi in the south eastern and Arokeke in the south western parts of Nigeria. It is used as vegetable in the preparation of many African dishes. In traditional folk medicine, the leaf is used for treatment of diabetes and hypertension as well as for treatment of typhoid fever (Okafor, 1987). It is also used to dispel stomach upset and pains and to enhance the return of menstrual cycle. Gongronema latifolium is primarily used as a staple vegetable/spice by some African cultures to help support the pancreas (Morebise et al., 2002). The hypoglycaemic and antihyperglycaemic properties of the ethanolic stem extract of Gongronema latifolium have been articulated in a review by (Farombi, 2003).
Garlic, Allium sativum L. is a member of the Alliaceae family, has been widely recognized as a valuable spice and a popular remedy for various ailments and physiological disorders. Cultivated practically throughout the world, garlic appears to have originated in central Asia and then spread to China, the Near East, and the Mediterranean region before moving west to Central and Southern Europe, Northern Africa (Egypt) and Mexico (Lutomski, 1987). Chinese strongly believe that garlic prolongs longevity (Srivastava et al., 1995) and is useful in treating most human diseases, including infections, cancer and heart diseases.
1.2 Statement of the Problem
This research investigate the comparative quantitative phytochemical analysis of ethanolic extracts of Gongronema latifolium and Allium sativum
1.3 Aim and Objectives
The aim of this research work is to determine the comparative quantitative phytochemical analysis of ethanol extracts of Gongronema latifolium and Allium sativum.
The objectives of this study include:
To investigate the phytochemical properties of G. latifolium only
To investigate the phytochemical properties of A. sativum only
To compare the quantitative phtytochemical of G. latifolium and A. sativum so as to justify its African ethnomedicinal use.
1.4 Significance of the Study
Gongronema latifollium plant is by herbalist in Nigerian folk medicine for the treatment of diabetes mellitus (Sakihama et al., 2002; Ugochwukwu et al., 2003 Edet etal., 2011). Also, a number of researchers have been carried out on the biological activity of the leaf extract of Gongronema latifolium. Ugochukwu et al. (2003) reported that the leaf extract of Gongronema latifolium is used for the treatment of non-insulin dependent diabetes mellitus (NNDM), investigated in the NIDDM-induced rats. Morebise et al. (2002); sakihama et al., (2002). Recent studies indicate that garlic extract has antimicrobial activity against many genera of bacteria, fungi and viruses. The role of garlic in preventing cardiovascular disease has been acclaimed by several authors. Chemical constituents of garlic have been investigated for treatment of hyperlipidemia, hypertension, platelet aggregation and blood fibrinolytic activity. Thus, will support it continued usage in nutrition, treatment and management of diseases.
1.5 Justification of the Study
Evaluation of plant products to treat diabetes mellitus is of growing interest as they contain many bioactive substances with therapeutic potential. In recent years, several authors evaluated and identified the ant-diabetic potential of Gongronema latifolium. A claim by herbalist proves that that Gongronema latifolium have been used to reduce blood sugar levels of diabetic mellitus patients. Also the study b Edet et al., (2011), which showed treatment of diabetes mellitus justify this investigation. Therefore, this study will identify the fractions of the plant that has more phytochemical properties.
1.6 Scope of the Study
This research work was specifically limited to the following;
Ø The preparation of the ethanolic extract of Gongronema latifolium
Ø The preparation of the ethanolic extract of Allium sativum
Ø Analysis of the phytochemical property of Allium sativum
Ø Analysis of the phytochemical property of Gongronema latifolium
1.7 Research Hypothesis
Null Hypothesis: There is no significant effect in the comparative quantitative phytochemical analysis of ethanolic extracts of Gongronema latifolium and Allium sativum
Alternative Hypothesis: there is a significant effect effect in the comparative quantitative phytochemical analysis of ethanolic extracts of Gongronema latifolium and Allium sativum
NUTRIGENOMICS
CHAPTER ONE
INTRODUCTION
The etiology of complex chronic diseases involves both environmental and genetic factors, with environmental influences such as diet exerting a greater effect among individuals with certain genetic profiles (David, 2005). Nutrition is clearly one of the most important determinants of health. Too much or too little of a nutrient can result in metabolic disturbances that predispose individuals to various diseases such as osteoporosis, diabetes, rheumatoid arthritis, cardiovascular disease (CVD) and certain types of cancer. Non-nutritive food bioactive can also affect the risk of developing various chronic diseases. Functional foods that are enriched with certain food bioactive have been suggested to play an important role in combating CVD and other chronic ill- nesses (David, 2005).
However, inconsistencies among epidemiological studies have yielded conflicting advice on the optimal level of intake for nutrients and specific food bioactive. These inconsistencies may be due, in part, to genetic difference between populations that are studied.
Nutrigenomics is the science that uses genomic information along with high-throughput ‘omics’ technologies to address issues important to nutrition and health (David, 2005).
Nutrigenomics is sometimes called nutritional genomics, which is increasingly being used as an umbrella term to refer to both the study of how diet affects genes and how genes affect diet (Kaptur et al., 2004).
One approach used to explore how dietary and genetic factors interact to influence various health outcomes is to examine how diet alters the function of genes or their protein products such as enzymes, receptors, transporters and ion channels that are known to regulate important biochemical pathways and cellular processes.
Another approach is to examine how variations in genes affect responsiveness to Specific dietary factors, an area that is sometimes referred to as nutrigenetics (David et al., 2005).
Candidate genes that are studied tend be those that are the targets of a nutrient or food bioactive, or those that impact the metabolism of the bioactive compound, including its absorption, biotransformation, distribution or elimination. For example, how efficiently we absorb fat, how rapidly we digest starch, or how slowly we eliminate caffeine from our circulation all determine the levels of a food bioactive that a target cell would be exposed to. Knowledge of the genetic basis for the variability in response to food bioactive should result in a more accurate measure of exposure of target tissues of interest to these compounds and their metabolites (David et al., 2005).
It has been demonstrated that numerous genetic polymorphisms can influence protein structure function. The Nutritional genomic area includes two parts: first Nutrigenomics that is the study of interaction between dietary components and the genome, and the regulating changes in proteins and other metabolism; second Nutrigenetics that identify the response to dietary components with regard to genetic differences (Subbiah, 2007). Nutrients are as environmental factors can interact with genetic material. It has been clearly demonstrated that DNA metabolism and repair depend on a wide range of dietary factors that act as cofactors or substrates in metabolic pathway, but much less is known about the impact of cofactors and/or micronutrients deficiency or excess on the fidelity of DNA replication and repair (Bull, 2008). Although the nutrients can influence the development of a particular phenotype, the response to a specific nutrient that determined by the individual genotype has also to be considered. The central role of genetic code in determining genome stability and related health outcomes such as developmental defects, degenerative diseases, and cancer is well-established (Fenech, 2008). The etiology of complex chronic diseases obviously relates to both environmental and genetic factors (El-Sohemy, 2007). Specifically, the “fetal basis of adult disease” or “early origins hypothesis” postulates that nutrition and other environmental factors during prenatal and early postnatal development influence gene expression and cellular plasticity, which can alter susceptibility to adult diseases (cardiovascular diseases, diabetes, obesity. etc) (El-Sohemy, 2007). The concept of nutrients effects on DNA stability, repair and on the different gene expression processes, recently became more prominent in nutritional science (Paoloni-Giacobino, 2003). Numerous dietary components can alter genetic and epigenetic events and therefore influence health (Trujillo, 2006). SNPs (single nucleotide polymorphisms) are the most common genetic variation, occur at about 500-2000 bp throughout the human genome, and normally found in at least 1% of the population (Ferguson, 2006). Many human studies have demonstrated the evidence for interaction between SNPs in various genes and the metabolic response to the diet. Moreover, SNPs analysis provides a potential molecular tool for investigating the role of nutrition in human health, diseases and identification of optimal diets (Ferguson, 2006). Nutrients and genome interact at two levels:
· Nutrients can induce or repress gene expression thereby altering individual phenotype.
· Conversely, single nucleotide polymorphisms can alter the bioactivity of important metabolic pathways and mediators and influence the ability of nutrients to interact with them
EFFECTS OF ETHANOLIC ROOT EXTRACT OF UGU OYIBO (Jatropha tanjorensis) ON HAEMATOLOGICAL INDICES OF FEMALE ALBINO WISTAR RAT
ABSTRACT
Jatropha tanjorensis is a natural medicinal herb which has been used over the years in the treatment of many health disorders. This study was carried out to determine the effects of ethanolic root extract of Jatropha tanjorensis on haematological indices of female albino wistar rats. Twenty (20) albino wistar rats weighing between 151-225 g were divided into 4 groups of 5 animals per group based on body weight. Group 1 served as the control and was fed with distilled water and normal rat feed. Group 2 was treated with 100mg of extract per kg body weight. Group 3 was administered with 250mg of extract per kg body weight while group 4 received 500mg of extract per kg body weight for 14 days. The haematological indices analysed include Red blood cells (RBC), White blood cells (WBC), Haemoglobin (HGB), Haematocrit (HCT), Mean cell volume (MCV), Mean cell Haemoglobin (MCH), Mean cell haemoglobin concentration (MCHC), Platelets count (PLT), Lymphocytes (LYM) and Neutrophil count. Result obtained showed that the extract caused an increase in White blood cell (WBC) count (12.28 1.32, 14.79 4.20, 16.80 1.20 for groups 2, 3 and 4 respectively), Red blood cell (RBC) count (7.65 0.22, 8.76 0.47, 9.29 0.48 for groups 2, 3 and 4 respectively) and Haemoglobin (HGB) count (15.34 0.54, 16.01 1.27, 17.01 1.49 for groups 2, 3 and 4 respectively). There were also significant (P 0.05) increase in the values of Haematocrit, Mean cell volume, Platelet count and Lymphocyte. The values of MCH and MCHC in group 2 were not significantly (P 0.05) different from the control while that of groups 3 and 4 were significantly (P 0.05) higher. For NEUT, groups 2 and 3 were not significantly (P 0.05) different from the control while group 4 was significantly (P 0.05) higher. The extract is said to boost the immune system of the treated animals and could prevent other opportunistic diseases associated with decreased immunity. Based on these findings, roots of J. tanjorensis may be important in the treatment and management of anaemia since they have positive effect on red blood cells (RBC) and haematocrit counts.
CHRONIC DISEASES AND THEIR PREVENTIVE MEASURES
CHAPTER ONE
INTRODUCTION
1.1 OVERVIEW
A chronic disease is a human health condition or disease that is persistent or otherwise long-lasting in its effects or a disease that comes with time. The term chronic is often applied when the course of the disease lasts for more than three months.
In Epidemiology, a risk factor is a variable associated with an increased risk of disease or infection for example tobacco smoking is a risk factor for lung cancer.
Preventive Measures consists of measures taken for disease prevention, as opposed to disease treatment.
Common chronic diseases include arthritis, asthma, cancer, COPD, diabetes and some viral diseases such as hepatitis C and HIV/AIDS. These diseases are generally preventable when risk factors are known and preventive measures are taken into proper consideration.
Chronic diseases constitute a major cause of mortality, and the World Health Organization (WHO) attributes 38 million deaths a year to chronic diseases (Ward and Black, 2016).
1.2 Types of Chronic Diseases
Chronic Disease have often been used to describe the various health related states of the human body such as syndromes, physical impairments, disabilities as well as diseases. Epidemiologists have found interest in chronic conditions due to the fact they contribute to disease, disability, and diminished physical and/or mental capacity. For example, high blood pressure or hypertension is considered to be not only a chronic condition itself but also correlated to diseases such as heart attack or stroke.
Additionally, some socioeconomic factors may be considered as a chronic condition as they lead to disability in daily life. An important one that public health officials in the social science setting have begun highlighting is chronic poverty (Hulme and Shepherd, 2003).
There are four major types of chronic disease classified by the World Health Organisation in 2015 and they are:
- Cancers
- Cardiovascular diseases, including cerebrovascular disease, heart failure, and ischemic cardiopathy
- Chronic respiratory diseases, such as asthma and chronic obstructive pulmonary disease (COPD)
- Diabetes mellitus
Other examples of chronic diseases and health conditions include:
· Addiction
· Alzheimer’s disease
· Atrial fibrillation
· Attention deficit hyperactivity disorder
· Autoimmune diseases, such as ulcerative colitis, lupus erythematosus, Crohn’s disease, coeliac disease, Hashimoto’s thyroiditis, and relapsing polychondritis
· Bipolar disorder
MODULATORY EFFECT OF ETHANOLIC LEAF EXTRACT OF SPHENOCENTRUM JOLLYANUM PIERRE (SPJ) ON LIVER FUNCTION AND HAEMATOLOGICAL INDICES OF CADMIUM CHLORIDE-INDUCED TOXICITY IN MALE ALBINO WISTAR RATS
ABSTRACT
The effect of ethanolic leaf extract of Sphenocentrum jollyanum pierre (SPJ) on liver function and hematological indices of cadmium chloride-induced toxicity in male albino wistar rats was investigated after 21 days of administration of the plant extract. Thirty (30) matured male wistar rats of albino strain were weighed and grouped into six (6) groups of five (5) animals per group according to their relative weight. Group 1 and 2 served as the negative and positive control group respectively. The animals in group 1 were not given the extract. Group 2 were given cadmium chloride at the concentration of 10mg/kg body weight. Group 3, 4, 5, and 6 served as the treatment groups receiving 200, 400, 200, and 400mg/kg of the plant extract for 21 days respectively. Group 3 and 4 were given 10mg/kg of cadmium chloride on the 22nd day. Administration of cadmium chloride (Cdcl2) at the concentration of 10mg/kgbw in group 2 (positive control) resulted in an increase in the mean serum AST value (243.75 ± 72.89) when compared to negative control (group 1) (161.00 ± 33.97). This is also visible in the mean serum ALT value (88.25 ± 29.69) when compared with the negative control (52.20 ± 12.01). There is a decrease in the mean serum AST value in group 3 and 4 (136.80 ± 33.00 and 77.50 ± 31.08) when compared to the positive control group having an increase mean serum AST value (243.75 ± 72.89). This is also same for mean serum ALT mean value (60.20 ± 23.62 and 44.25 ± 11.98) in group 3 and 4 when compared to the positive control group (88.25 ± 29.69). In haematological evaluation, white blood cell was not significantly altered. The effect of the extract in the final body weight and absolute liver weight of the experimental animals were also observed in the treatment groups when compared to the negative control group
Keywords: Shenocentrum jollyanum pierre, cadmium chloride, wistar rats, liver function.
CHAPTER ONE
INTRODUCTION
1.1 Background of the Study
The use of medicinal plants as a substitute for orthodox drugs in the management of various diseases has been increasing globally due to the unavailability of modern health facilities, relative availability of medicinal herbs and recent revelations that they possess active compounds that may be responsible for different biological and pharmacological actions (Tiwari and Mehta, 2013).
Herbs and herbal formulations for the treatment of ailments have continued to receive increased attention because of the strong belief that these products are safe (Farnsworth and Soejarto, 1985; Said et al., 2002). This assumption to a large extent may have influenced the indiscriminate use of these formulations by many, particularly amongst the rural populace. The incidence of adverse effects and sometimes life-threatening conditions allegedly emanating from these herbal medicines has been reported among various ethnic groups (Elvin-Lewis, 2001; Chan, 2003). Consequently, it has become imperative to ascertain the toxicity profile of these medicinal herbs.
According to a World Health Organization (WHO) report, it is estimated that close to 80% of the people living in the third world nations of the world depend on traditional and complementary medicines for their basic health care.
Sphenocentrum jollyanum Pierre (Menispermaceae) is a perennial plant that grows naturally along the west coast sub-region of Africa with expanse from Cameroon across Nigeria to Sierra Leone (Mbaka et al, 2010). It occurs in undergrowth of dense forest, which thrives in deep shade from sea-level up to 400m altitude (Nia et al, 2004). It is mostly found in regions with mean annual rainfall of 1800 mm or more, mean minimum temperature of 20◦C, and mean maximum of 29◦C.
Sphenocentrum jollyanum pierre is a medicinal plant which has been shown to display a wide spectrum of biological and pharmacological activities. The plant has various parts which contain wide array of phytochemicals that has medicinal values. From the existing knowledge, there is no literature on its toxicity profile. Its medicinal importance was first reported by Dalziel (1955) in which it was noted that the leaves decoctions were used as vermifuge.
Many scientific work has been carried out on this plant in relation to his antioxidant property (Nia et al, 2004), antidiabetic role (Mbaka et al, 2011), antiviral and anti-inflammatory activity (Moody et al, 2006), wound healing property, hepatoprotective effect (Olorunnisola et al, 2011), sexual stimulant activity (Owiredu et al, 2007) etc.
Sphenocentrum jollyanum pierre contains phytochemicals which include saponins, flavonoids, alkaloids, terpenoids, anthraquinones, tannins (Amidu et al, 2008; Woode et al, 2009; Mbaka et al, 2011).
1.2 Statement of Problem
Sphenocentrum jollyanum pierre extracts has a broad spectrum of biologic and pharmacological activities. The extract obtained from the various parts of the plants are used in the management of various health conditions owing to the phytochemicals present which confer its therapeutic effect. For instance, normal levels of biochemical parameters (such as alkaline phosphatase {ALP}, aspartate aminotransferase {AST}, alanine aminotransferase {ALT}, bilirubin) in serum may reflect the normal functioning status of a body’s organ such as the liver but changes in the levels of these biochemical parameters greater than or less than the normal reference range is a useful indicator of the diseased state of the liver. The liver biochemical parameters measured are being released into the blood when there is a damage to the cell membrane of the liver . Measurement of these biochemical parameters are of value in the diagnosis and management of wide variety of diseases of the liver. Hence, this study was designed to ascertain the effects of Sphenocentrum jollyanum pierre leaves extract on liver biochemical parameters and also on haematological indices of male albino wistar rats after administration of cadmium chloride as a known toxicant.
1.3 Aims and Objectives
To ascertain the effect of Sphenocentrum jollyanum pierre (Menispermaceae) leaf extract on liver function and haematological indices of cadmium chloride induced toxicity in male albino wistar rats.
1.4 Significance of the Study
Humans on daily basis however are exposed to xenobiotics and thus try to eliminate them from the body system by means of metabolism. The liver and kidney are responsible to metabolized these xenobiotics and can itself be damaged or its integrity compromised. This can pose a serious health threat or complications including liver failure, kidney failure and in a more severe case cause death.
Sphenocentrum jollyanum pierre is primarily used in Nigeria and other tropical regions of Africa for medicinal purposes to treat several diseases and has also been suggested to exert protective medicinal effect on the liver and the kidney.
This study “effects of Sphenocentrum jollyanum pierre leaves extract in cadmium chloride-induced toxicity in male albino wistar rats” was designed to investigate and justify that the plant extract has a protective medicinal effects on the functions of the liver and also on haematological parameters. It also helps to know the proper doses and treatment for certain diseases.
1.5 Scope of Study
The scope of work covered the ascertainment of the following:
1. Extraction of crude extract of the leaf of Sphenocentrum jollyanum pierre with 80% ethanol.
2. Determination of LD50 of the plant extract.
3. Preparation of the stock solution of the plant extract
4. Preparation of the administered dose of the plant extract per body weight of the animals.
5. Preparation of cadmium chloride solution.
6. Dissection of the rats at the end of the administration to obtain blood sample for biochemical analysis of the liver, haematological analysis, and collection of organs (liver) for tissue histology.
7. Determination of the effect of ethanol extract of the leaves of the plant on liver biochemical parameters, haematological indices, and the organs.
8. Data analysis and calculation.
9. Statistical analysis.
1.6 Hypothesis
The following hypothesis serves as a guide to this study.
Null Hypothesis: There is no significant diferrence (p>0.05) on the parameters analysed after administration of ethanol extract of the leaves of Sphenocentrum jollyanum pierre against cadmium chloride-induced in adult male wistar rats of albino strain.
Alternate Hypothesis: There is a significant difference (p<0.05) on the parameters analysed after administration of ethanol extract of the leaves of Sphenocentrum jollyanum pierre against cadmium chloride-induced toxicity in adult male wistar rats of albino strain.
ETHANOL LEAF EXTRACT OF Irvingia gabonensis O’Rorke Baill MITIGATES SODIUM ARSENITE-INDUCED LIPID PROFILE DERANGEMENTS IN WISTAR RATS
ABSTRACT
The effect of ethanol leaf extract of Irvingia gabonensis O’Rorke Baill on lipid profile in sodium arsenite-exposed Wistar albino rats was investigated. Forty (40) female wistar rats of weights between 100g-161g were randomized into 8 groups of 5 animals each. Group 1 served as control and was given normal feed and water ad libitum. Group 2 was administered 4.1mg/kg body weight (mg/Kgbw) sodium arsenite (SA) for 14days. Groups 3, 4, and 5 were post-treated with 100mg/Kgbw, 200mg/Kgbw, and 400mg/Kgbw of extract respectively for 14 days after SA intoxication for 14 days. Groups 6, 7, and 8 were administered 100mg/Kgbw, 200mg/Kgbw, and 400mg/Kgbw of extract respectively for 14days. Treatments were by oral administration and lasted for 28days. Sodium arsenite toxicity was shown by significant (p<0.05) increases in serum Total Cholesterol (TC), Triacylglycerol (TAG), High density lipoprotein (HDL), Low density lipoprotein (LDL), Very low density lipoprotein (VLDL), and total lipid levels in group 2 as compared with the control. However, treatment with extract at different doses (groups 3, 4, 5) led to significant (p<0.05) decreases in all assayed parameters compared with group 2. Furthermore, treatment with extract only at different doses showed significant (p<0.05) and non-significant (p˃0.05) increases in assayed parameters compared with control except LDL levels which decreased significantly. Ethanol leaf extract of Irvingia gabonensis O’Rorke Baill may therefore be very useful in mitigating sodium arsenite-induced lipid metabolism derangements in Wistar rats