Abstract

The aim of this study was to determine the pancreatic amylase activity of 24 female Wistar rats fed with cassava- cyanide diet and as well as the in-vivo effect of Malus domestica and Moringa-oleifera seed. Photochemical constituent of Moringa-oleifera have proven to have hypotenstive, anticancer and antibacterial activity and in Malus domestica it is reported to have very strong antioxidant and lower cholesterol. The result indicates the assay for pancreatic amylase which were determined to be 109.5± 2. 08 for control group 1a following 110. 76±2.06 µL-1 control 1b others were 122.50 ± 3.0 µL-1, 11 7.75± 2.06 µL-1 and 120.0±2.16 µL-1 for groups 2 to 5 respectively. Group 3 and 5 had the highest enzyme activity mean which show that pancreatic amylase was affected when test groups were fed with diets containing cassava- cyanide but Malus domestica and Moringa-oleifera seed reduced the cyanide in the pancreases in the decreasing order Moringa-oleifera seed is greater than Malus domestica.Pancreatic amylase assay was done as describe by Agape Kit.

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CHAPTER ONE

INTRODUCTION


1.1   Background to the Study

Cyanide is highly toxic to living organisms particularly in inactivating the respiration system by tightly binding to terminal oxidase (Poter, 1983). However, large amounts of cyanide are used in industries involved in the metalplanting pharmaceutical synthetic fibers, plastics. Amylase are a group of hydrolases that degrade complex carbohydrates into fragements. Amylase is produced by the exocrine pancreas and the3 salivary glands to aidin the digestion of starch. It is also produced by the small intestine mucosa, ovaries, placenta, liver and falloplan tubes.(sternby, 1996).

The pancrease is located behind the stomach and have two very important functions first it helps produce digestive enzymes that help in digestion of carbohydrate, protein and fats second, it produces very important hormones called the in such and glucagon. These hormones are involved in regulation of blood sugar levels, without which the sugar levels in the body will go for a toss. When the pancreas become inflamed, it leads to pancreatis (Moss 1999).

Apple (Malus domestica) has been revealed that when consumed is medicinal and can provide antiproliferative, anticarcinogenic, anti-inflammatory and lower incidence of lung cancer, viral diseases and cardiovasauar disorder amongst other health benefits associated with its high anti-oxidant (Sara, 2012) and also Moringa-oleifera has it phytochemical which treats hypotensive anticancer , and antibacterial activity (Jed W. Fahey, 2005).

1.2    Aims and Objectives

This experiment was carried out to determine the effect of cyanide on pancreatic amylase and to determine the efficacy of Malus domestica and moringa oleifera as supplements to reduce the effect of cyanide pancreatic amylase activity.

1.3    Literature Review

1.3.1 Food Cassava: Cassava

Cassava (manihot esculenta crantz) a dicotyledonous plant of the Euphorbiaceae family, is an extensively cultivated plant in many tropical regions. Its drought tolerance and ability to various soil and weather conditions makes it a choice staple root crop for farmers (Madukosiri and Amos Tautua 2010) cassava is originated in the Americas. It is a shrub with an average height of one metre and has a palmate leaf formation. Cassava produces bulky storage roots with a heavy concentration of carbohydrates about 80%. The shoot grow into leaves that constitute a good vegetable rich in proteins, Vitamins and Mineral cassava leaves and roots, if properly processed, can therefore provider a balance diet protecting millions of Africa children against Malnutrition. (Babalolo Temilade, 2010).

1.3.2 Types of Cassava

There are two types of cassava the sweet and the bitter, the sweet varieties contain considerably less linamarin that the bitter type.

1.3.3  Nutritional Profile.

Cassava tuber has high carbohydrate content, but is not a good source of protein it contains vitamin C, potassium and dietary fibre. Cassava leaves have higher protein content, contain vitamin C and Vitamins A and provide some dietary fibre.

1.3.4   Anti Nutrients of Cassava

Anti- nutrients is are natural or synthetic compounds that interfere with the absorption of nutrients. One common example is phytate which forms insoluble complexes with calcium zinc Inn and copper. Protein can also be anti-nutrients, such as the tyrosine inhibitors and lectins found in legumes (Gilani et al, 2005).

Another particular widespread form of anti-nutrient are flavonoids, which are a group of polphenolic compound that includes tannins. They also inhibit digestive enzymes and precipitate proteins (Beecher, 2003).

1.3.5  Chemical and Physical Information of Cyanide

The term cyanide refers to any compound that contains the cyanide ion (CN-) consisting of a carbon atom. Triple bonded to a nitrogen atom. Hydrogen cyanides (HCN) is colorless or pale blue liquid or gas with faint bit#ter almond –like odor, while sodium cyanide (NaCN) and potassium cyanide (KCN) are white crystalline powders. HCN is a weak acid with a pka of 9.2 therefore, HCN and CN can inter convert based on Ph and temperature. Isolation under physiological conditions, the majority of HCN is present in the undissociated form.

The dissociation constants of metallocyanides vary significantly depending on oxidation states, PH temperature and photo degradation (Beck, 1987). Cyanide is also a component of tobacco smoke and can be present at high concentrations in structural fives (sternumaus, 2007).

1.3.6  Absorption

The available data show that cyanide is rapidly and extensively absorbed via the oral, inhalation and dermal routes, although quantitative data on the percent or extent of absorption are limited oral absorption has been reported as being lower at lethal does.
Some cyanide salts, including potassium cyanide (KCN) and sodium cyanide (NaCN), rapidly dissociate in water.

1.3.7  Distribution

Cyanide distributes rapidly and uniformly throughout the body following absorption. HCN enters the systemic circulation when inhaled or dermally absorbed (Yamamoto 1982). Limited qualitative and quantitative data care available regarding the tissue distribution of cyanide in humans from inhalation exposure to high doses of cyanide Example, cyanide was found in the lung, heart kidney pancreases gland who died following cyanide in halation (Gettler 1938).

1.3.8   Metabolisms

The major metabolism pathway for cyanide is conversion to the less acutely toxic compound, thiocyanate primarily by rhodanese, with some conversion occurring via 3- mercaptopyruvate sulfur transferees conversion to thiocyanate accounts for 60-80% of a cyanide dose.

Minor pathways include incorporation in a 1-carbon metabolic pool 0r conversion to 2-aminothiazoline-4-carboxylic acid (ATSDR, 2006).

Conversion to 2-aminothiazoline-4carboxylic acid via reaction with cystine accounted for approximately 15% of an injected dose of cyanide in rats (Wood and Cooley, 1956).

1.3.9 Metabolism pathway of cyanide

CN major path (80%) Thiocyanate Urinary excretion
Cyanide (SCN)

Note: Details here are fully represented in full material

1.3.9 Elimination

Data in human and animals indicate that cyanide is primarily excreted in the Urine as thiocyanate following both inhalation and oral exposure smaller amounts are excreted as culinary cyanide or as HCN or carbon dioxide in exhaled air. (Chandra et al, 1980).

1.3.11 Effect on Enzyme and other Biochemical parameter

Hydrogen cyanide in a activities the enzyme cytochrome oxidase in the mitochondria of cells by binding to the Fe3+ /fe2+ contained in the enzyme. This causes a decrease in the utilization of oxygen if the tissues cyanide causes an increase in blood glucose and lactic acid levels and a decrease in the ATP/ADP ratio indicating a shift from aerobic to anaerobic metabolism. Cyanide activates glycogenolysis and shunts glucose to the pentose phosphate pathway decreasing the crate of glycolysis and inhibiting the tricarboxylic and cycle (Oke,1980)

1.3.12 Pancrease

The pancreas is located behind the stomach and have two very important functions. First it helps produce digestive enzymes that help in digestion of carbohydrate, protein and fats. Second ,it produces very important hormones called the insulin and glucagon. These hormones are involued in regulation of blood sugar levels, with out which the sugar levels in the body will go for a toss. When the pancreas become inflamed, it leads to pancreatis. Which has 2 types, Acute and Chronic pancreatitis( Puylaert 2011).

Fig 1.1 Pancreas (Erigi 2014)

Pancreas is the hydrolysis of starch by saliva, due to the presence of an enzyme is saliva “ptyalin” an amylase (puylaert M, 2011).

1.3.13 Function of Pancreas

To make digestive enzymes which help us to digest food. Enzymes are special chemicals which help to speed up your body’s processes. To make hormones which regulate our metabolism.

1.3.14 Metabolic Role of Pancreas Hormones:

Hormones are chemicals which can be released into the blood stream. They act as messengers affecting cells, and tissues in distant parts of your body. About 90% of the pancreas is dedicated to making digestive enzymes cells called acinar cells within the pancreas produce. The enzyme help make proteins, fats and carbohydrates smaller. This helps the intestines to absorb these nutrients. The acinar cells also make a liquid which creates the right condition for pancreatic juice.

1.3.15 Enzymes involved in pancreas Hormones

⦁ Pancreatic proteates (such as trypsin and chymotrypsin). Which help to digest proteins
⦁ Pancreatic amylase- which helps to digest carbohydrates (sugars).
⦁ Pancreatic lipase- which helps to digest fat.

Approximately 5% of the pancreas makes hormones which help to regulate your body’s metabolism. These hormones are made by several different cells which clump together like little islands (islets) within the pancreas. The islets are called islets of langerhans and there are about one million islet dotted about in a n pancreas hormones made by the cells in the islets of langerhans within the pancreas include:

⦁ Insulin- which helps to regulate sugar levels in the blood
⦁ Glucagon- which works with insulin to keep blood sugar levels balanced
⦁ Somatostatin- which helps to control the release of other hormone
⦁ Gastric- which aids digestion in the stomach.
1.5.4 Disorders of the pancreas

Ductal adenocarcinoma of the pancreas is a type of cancer that is develop from a cell which becomes cancerous in the pancreatic duct. This multiples and a tumor then develop in and around the duct. As the tumor enlarges from other types of cells within the pancreases are insulinamas and glucagonomas (Dragovich, 2011).

1.3.16 Causes of Pancreatic Cancer

A cancerous tumor starts from one abnormal cell. What seems to happen is that certain vital genes which control how cells divide and multiply are damaged or altered which makes the cell abnormal. If the abnormal cell survives it may multiply out of control into a malignant tumor. Other causes include Diabetes, aging, smoking, obesity, chemicals; heavy exposure at work to certain pesticides, dyes and and chemicals used in metal refining may increase the risk.

1.3.17 Photochemical Characterization on Apple and Moringa

Recent studies has reveal that dietary intake of selected plant- fruits, vegetables and seeds, especially those containing functional bioactive compounds such as phenolic acids, tannins, flavonoids, vitamins and essential minerals, is linked to reduced prevalence of oxidative stress associated diseases such as cardiovascular diseases, cancer and other health disorders.

1.3.18 Medicinal Plant: Moringa Oleifera

Moringa oleifera Lam (also called ”The Miracle Tree,” “Horseradish-tree,” or “Ben oil tree”) is the best known and most widely distributed species of Moringaceae family, having an impressive range of medicinal uses with high nutritional value throughout the world (Suaib et al, 2011). Analyses of the proximate composition of M. oleifera seeds have showed high levels of vitamins, minerals, lipids and essential amino acids with minor variations depending on climatic conditions, time of the year and different soil types from which the seeds were collected (Abdulkarim et al, 2005). In regard to anti-nutritional factors, the seeds contain an acidic protein with hemagglutinating activity, glucosinolates (65.5µmol/g) and phytates (41g/kg).

Phytates present to an extent of 1% to 6% reduce mineral bioavailability in monogastric animals, particularly, Zn2+ and Ca2+. Lectins, on the other hand, are usually responsible for agglutinating cells, interacting with intestinal epithelium, interfering with nutrient digestion and absorption and reducing food efficiency (Paulo et al, 2008). Specific phytochemical in Moringa that have been reported to have hypotensive, anticancer, and antibacterial activity includes: 4-(4′-O-acetyl-α-Lrhamnopyranosyloxy) benzylisothiocyanate,4-(-L rhamnopyranosyloxy) benzylisothiocyanate, niazimicin, pterygospermin, benzyl isothiocyanate, and 4-(α-L-rhamnopyranosyloxy)benzyl glucosinolate (Jed, 2005).

1.3.19 Apple and its Health Benefits

Apple (Malus domestica Borkh.), a member of the family Rosaceae, is the fourth most frequently consumed fruits in many regions across the World. Apple fruit has been identified as an excellent potential source of carbohydrates, vitamins, minerals, dietary fiber and phytonutrients. Recent studies revealed that consumption of apple fruit is medicinal and can provide antiproliferative, anti-carcinogenic, anti-inflammatory effects and lower incidence of lung cancer, viral diseases, and cardiovascular disorders amongst other health benefits associated with its high antioxidant. The antioxidant capacity of apple is mostly attributed to presence of vitamins C, caroteniods, phenolic compounds and other phytonutrients. (Pugalenti et al, 2012).

However total antioxidant activity of apples with the peel as determined by Vaibhav et al, (2012) was approximately 83μmol vitamin-C equivalents, which means that the antioxidant activity of 100g whole apples is equivalent to about 1500 mg of vitamin-C but the amount of vitamin-C in 100 g of apples is only about 5.7 mg (less than 4% of total apple antioxidant activity) implying that all of the antioxidant activity from apples comes from a variety of other phytochemicals as asserted by (veronica et al, 2002). The prominent phenolic compounds in apple fruit includes: Quercetin, Catechin, Phloretin, gallic acid and Chlorogenic acid (Vaibhav et al, 2012).

1.3.20 Overview on Some Phytochemicals

1.7.1 Flavonoids:

Flavonoids are one class of secondary plant metabolites that are also known as Vitamin P or citrin. These metabolites are mostly used in plants to produce yellow and other pigments which play a big role in colouring the plants. In addition, Flavonoids are readily ingested by humans and they seem to display important anti-inflammatory, anti-allergic and anti-cancer activities. Flavonoids are also found to be powerful anti-oxidants and researchers are looking into their ability to prevent cancer and cardiovascular diseases. Flavonoids are synthesized by the phenylpropanoid metabolic pathway where the amino acid phenylalanine is used to produce 4-coumaryol-CoA, and this is then combined with malonyl-CoA to produce chalcones which are backbones of Flavonoids. Chalcones are aromatic ketones with two phenyl rings that are important in many biological compounds (Crozier et al, 2006).

1.7.2 Saponins

Saponins are glucosides with foaming characteristics. Saponins consist of a polycyclic aglycones attached to one or more sugar side chains. The aglycone part, which is also called sapogenin, is either steroid (C27) or a triterpene (C30). The foaming ability of saponins is caused by the combination of a hydrophobic (fat-soluble) sapogenin and a hydrophilic (water-soluble) sugar part. Saponins have a bitter taste. Some saponins are toxic and are known as sapotoxin. The non-sugar part of saponins has also a direct antioxidant activity. Saponins are used widely for their effects on ammonia emissions in animal feeding (Thakur et al, 2011).

1.7.3 Tannins:

Tannins are common in fruits (grapes, persimmon, blueberry, etc.), in tea, in chocolate, in legume forages (trefoil, etc.), in legume trees. Tannins are astringent, bitter plant polyphenolic compound of high molecular weight ranging from 500 to over 3000, containing sufficient hydroxyls and other suitable groups (i.e. carboxyls) to form effectively strong complexes with protein and other macromolecules including carbohydrates. They are biosynthesized via the shikimic acid or phenylpropanoid pathway and stored in the vacuoles or surface wax of plants especially the Gymnosperms and Angiosperm (Peter and van, 2013).

 

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