Dr. Onkar S.

Bains BISC 313 SFU Spring 2013

 Metals are probably the oldest toxins known to humans

Lead usage may have begun prior to 2000 BC in the smelting of silver Arsenic was obtained during melting of copper and tin, and an early use was for decoration in Egyptian tombs

Introduction

The five main heavy metalsmercury (Hg), lead (Pb), cadmium (Cd), chromium (Cr), and arsenic (As)present the greatest environmental hazard due to their extensive use, their toxicity, and their widespread distribution

 Iron for instance forms an essential part of hemoglobin, a protein in our blood which transports oxygen from the longs to other tissues. Although necessary, essential metals become toxic at high concentrations.

Non-essential heavy metals include lead (Pb), mercury (Hg), cadmium (Cd) and tin (Sn)
They can be tolerated at low levels, but become toxic as well at higher concentrations

QUALITY OF HEALTH

Heavy metals consist of both essential and non-essential Biological essential heavy metals include copper (Cu), nickel (Ni), iron (Fe) and zinc (Zn)

DOSE OF METAL

 "Heavy metals" are chemical elements with a specific gravity at least 5 times that of water
The specific gravity of water is 1 g/cm3 at 4C (39F) Some well-known toxic metals with a specific gravity five or more times that of water are arsenic (5.8), cadmium (8.7), chromium (7.2), lead (11.3), and mercury (13.5)

In small quantities, certain heavy metals are nutritionally essential for a healthy life
Some of these are trace elements (i.e., iron, copper, manganese, and zinc)
These elements, or some form of them, are commonly found naturally in foodstuffs, fruits and vegetables, and in commercially available multivitamin products

 Heavy metals become toxic when they are not metabolized by body and accumulate in soft tissues

Heavy metals may enter human body via food, water, air, or absorption through the skin in agriculture, manufacturing, pharmaceutical, industrial, or residential settings

Factors affecting toxicity of metals

Alter function of essential metals
Toxic metals may displace essential metals as co-factors for enzymes

Lifestyle factors
Smoking contains toxic metals such as cadmium and arsenic

Formation of metal complexes

Metallothioneins form complexes with cadmium, zinc, copper and other metals by interacting with sulfhydryl groups on cysteine residues Captured metals are transported to kidney for filtration and excretion

 Chemical form/speciation
The free metal ion is considered most bioavailable and hence the most toxic
Example: Metal sorption by soil (organic matter, clay minerals) makes metal ions less bioavailable Example: High pH decreases bioavailability (metals predominantly found an insoluble meta mineral phosphates and carbonates) while low pH increases bioavailability (commonly found as free ionic species or as soluble organometals) Example: Under oxidizing or aerobic conditions, metals found as soluble cationic forms (i.e., Cd2+ or Pb2+) while in areas rich in sulfur and sulfatereducing bacteria, the sulfide that is generated is available to form nontoxic, insoluble sulfide deposits (i.e., CdS or PbS)

Lipid soluble and non-ionized pass membrane easily (i.e., tetraethyl lead and methylmercury) Some metals are more toxic in one speciation state than another (i.e., chromium VI is more toxic than chromium III)

 Age
Young children and elderly more susceptible to metal toxicity Major exposure of many toxic chemicals in children is food (children consume more calories per pound of body weight than adults) Also, children have increased GI tract absorption with respect to metals, particularly lead Newborns have increase exposure of toxic metals from breast milk contaminated with lipophilic metals

Immune status of host

Metals shown to provoke immune reactions (some examples of metals that can do this are mercury, chromium, platinum beryllium and nickel)

General mechanisms of toxicity with heavy metals

Protein inactivation
Can substitute in for essential metals that are involved in the binding of substrate to active site of enzymes Bind to sulfhydryl group (SH) found within proteins (thereby prevent disulfide bridge formation) Example: Microtubule function disruption (affect chromosomal separation during cell division or affect cilia/flagella movement)

Oxidative stress
Reflects an imbalance between systemic manifestation of reactive oxygen species and a biological system's ability to readily detoxify reactive intermediates or to repair resulting damage Heavy metals disturb redox homeostasis by stimulating formation of reactive oxygen species such as superoxide anions and hydroxyl radicals

Interference with DNA transcription, translation and repair

Heavy metals to cover in course

Mercury Cadmium Arsenic Lead will mention at next lecture

 Mercury is found in many rocks including coal

Mercury

When coal is burned, mercury is released into the environment

Mercury that was once in the Earths crust could be released through a volcanic eruption or other geological activity Mercury can also be released to environment by human activities such as metal smelting, iron and steel production, coal-fired electricity generation, industrial boilers, cement kilns, waste incineration, and use of products such as electrical switches and fluorescent lights

Chemical forms of mercury

Elemental/metallic: Hg0
A metallic, silvery liquid (also referred to as quicksilver) that is processed from an ore called cinnabar Readily breaks into droplets and easily vaporizes at room temperature into an odorless, colorless vapor that can easily be inhaled High absorption from lungs (~80%) but poorly absorbed from GI tract (~20%) Readily absorbed due to its lack of charge and is highly lipid soluble It easily crosses cell membranes and barriers (i.e., blood-brain, placental), and becomes trapped in cells when it is reduced to Hg1+ or Hg2+

Inorganic salts: Divalent or mercuric (Hg2+), Monovalent or mercurous (Hg1+)

Includes complexes: examples include HgX2, HgX3-, HgX42- (X=OH-, Cl-, Br-) Low absorption because these positively charged ions do not readily transverse cell membranes

Chemical forms of mercury

Organic: CH3Hg+, CH3CH2Hg+, (CH3)2Hg
Alkyl groups added to mercury can be dimethyl, ethyl, or methyl substituents High absorption from GI tract (~90-95%)...also readily absorbed through skin Absorption due to being highly lipid soluble Can cross blood brain and placental barrier using methionine transport system (molecular mimicry)
When methyl-Hg bind to SH group on cysteine, the molecule looks similar to methionine and takes its place in the membrane transport system which normally moves methionine into the brain or fetus

Able to easily bioaccumulate in organisms and biomagnify up a food chain

+ cysteine

methylmercury methionine

Biomagnification of methylmercury

Environmental cycling of mercury

and biomagnification

Hg2+(major dissolved form in freshwater), HgCl42-(major dissolved form in seawater

 Biomethylation
Process whereby living organisms produce a direct linkage of a methyl group to a metal, thus forming metal-carbon bonds Organisms involved are as follows:
Sulphate-reducing bacteria found in anoxic waters and sediments Some aerobic and facultatively anaerobic bacteria Fungi and lower algae Plants, animals and humans

Demethylation
Process whereby methyl group is removed from a molecule Example: bacterial enzymes efficiently transform the most toxic forms of mercury to less toxic states

Methylmercury lyase (MerB) acts on methylmercury to release less toxic ionic mercury Hg2+ Mercury reductase (MerA) acts on Hg2+ to less toxic volatile metallic Hg0

Mad hatter syndrome

Chronic mercury poisoning among hatmakers whose felting work involved prolonged exposure to mercury Mercury was commonly used in the manufacture of felt hats, especially in the 19th century
Once the fur has been rolled, beaten, and treated with mercury, it is called felt

Victims developed severe and uncontrollable muscular tremors and twitching limbs, called "hatter's shakes"; other symptoms included distorted vision and confused speech Advanced cases developed hallucinations and other psychotic symptoms

 Minamata Bay region on the island of Kyushu, Japan The Chisso Corporation was once a fertilizer company, and gradually advanced to a petrochemical and plastic-maker company From 1932 to 1968, an estimated 27 tons of mercury compounds was dumped into Minamata Bay Mercury accumulated in sea creatures, leading eventually to mercury poisoning in human population

Case: Minamata Bay poisoning

Fish and shellfish consumption resulted in exposure to high levels of methylmercury Microorganisms biotransformed mercury into methylmercury

In 1952, the first incidents of mercury poisoning appear in the population of Minamata Bay in Japan, caused by consumption of fish polluted with mercury, bringing nearly 1000 fatalities

 Hundreds of people exhibited serious neurological problems such as:

Difficulty walking, swallowing, speaking, and hearing Post mortem brain analysis revealed that many had a marked loss of brain weight and volume (brain atrophy) Children born to exposed mothers had a high rate of birth defects, including severe brain damage, mental impairment, and delayed development

 Mass methylmercury poisoning incident that began in late 1971 Iraq was rocked by a severe drought in 1971 which led to ubiquitous food shortages Excess grain in the United States and Mexico, which had been treated with a methylmercury fungicide, was shipped as aid
Grain had distinctive orange-pink color

Case: Iraq poison grain disaster

People suffered from parathesia (numbness of skin), ataxia (lack of coordination of muscle movements) and loss of vision, symptoms similar to those seen when Minamata disease affected Japan The recorded death toll was 650 people, but figures at least ten times greater have been suggested

Cadmium
Occurs in nature primarily in association with lead and zinc ores and is released near mines and smelters processing these ores Industrially cadmium is used as a pigment in paints and plastics, in electroplating, and in making alloys and alkali storage batteries (e.g., nickelcadmium batteries) Environmental exposure to cadmium is mainly from contamination of groundwater from smelting and industrial uses as well as the use of sewage sludge as a food-crop fertilizer

 McDonald's recalls cadmium-tainted Shrek glasses

In June 2010 12 million collectibles recalled after tests reveal cadmium in paint Cadmium is a known carcinogen that research shows also can cause bone softening and severe kidney problems Potential danger would be long-term exposure to low levels of cadmium, which could leach from paint onto child's hand, then enter the body via oral route of administration if hands are left unwashed

 Cadmium is present in circulatory system bound primarily to metallothioneins, produced in the liver
Cadmium + metallothionein = CdMT

Cadmium toxicity

Main organ of damage following longterm exposure is the kidney, with the proximal tubules being primary site of action Following glomerular filtration in kidney, CdMT is re-absorbed efficiently by proximal tubule cells, where it accumulates within lysosomes Subsequent degradation of the CdMT complex releases Cd2+, which inhibits lysosomal function, resulting in tubule cell injury

 Kidneys lose their function to remove acids from the blood in proximal renal tubular dysfunction
Decreased H+ secretion into renal filtrate

This dysfunction causes gout, a form of arthritis due to the accumulation of uric acid crystals in joints because of high acidity of blood (hyperuricemia)

The proximal renal tubular dysfunction also creates low phosphate levels in the blood (hypophosphatemia), causing muscle weakness and sometimes coma
Decreased phophate reabsorption Low phosphate levels also linked to poor bone mineralization

 The proximal renal tubular dysfunction also will affect conversion of 25(OH) vitamin D to 1,25-(OH)2 vitamin D this will in turn decrease calcium reabsorption at small intestine and decrease bone mineralization due to low serum calcium levels

Case: Itai-itai disease in Toyama Prefecture

Located in the Hokuriku region on Honshu island of Japan Mining was prevalent in the Toyama Prefecture of Japan starting around the year 710 After World War I, new mining technology arriving from Europe made the Kamioka Mines in Toyama among the most productive in the world Starting all the way back in 1910 cadmium was being released in significant quantities into the Jinzu River in Toyama

 This was a major problem because cadmium in the water killed all fish, not to mention it was the major source for irrigation for the surrounding paddy fields, as well as drinking water In 1912, first documented case of disease emerged Itai-Itai resulted primarily from consumption of cadmium contaminated rice From 1939 to 1954, ~200 people affected Itai-itai disease literally translates to ouch-ouch disease, named for the painful screams of its victims Major symptoms of this disease:
Osteomalacia (softening of the bones) Osteoporosis (loss of bone mass and weakness) Kidney failure

Arsenic
Arsenic is a naturally occurring element widely distributed in the earth's crust Occurs naturally in soil and minerals and may enter the air, water, and land from wind-blown dust and may get into water from runoff and leaching Man-made sources:
Smelting of gold, silver, copper, lead and zinc ores Combustion of fossil fuels Agricultural uses as herbicides and fungicides Cigarette smoke Occupational: largest source is manufacture of pesticides and herbicides

 Elemental/metallic: As0

Chemical forms of arsenic

Element is a steel gray, very brittle, crystalline, semi-metallic solid

Readily absorbed due to its lack of charge and is lipid soluble It easily crosses cell membranes and barriers

Inorganic salts: primarily in two oxidation states (+3 to +5)

Arsenic acid: AsO(OH)3 Arsenous acid: As(OH)3 Arsine, AsH3, an extremely poisonous gas
Rapidly absorbed through the lungs

Arsenous acid

Organic:
Arsenic can be methylated by bacteria, algae, fungi, vascular plants, and animals Methylated arsenic (V) compounds include: Monomethylarsonic acid, CH3AsO(OH)2; Dimethylarsinic acid, (CH3)2AsOOH; Trimethylarsine oxide, (CH3)3AsO Methylated arsenic (III) compounds include: Monomethylarsine, CH3AsH2; Dimethylarsine, (CH3)2AsH; Trimethylarsine, (CH3)3As Absorption due to being highly lipid soluble

Arsenic toxicity
Ranking of toxicity: organic arsenics > inorganic arsenics > elemental Trivalent forms more toxic than pentavalent forms Acute exposure: severe abdominal pain, fever, cardiac arrhythmia Chronic exposure: muscle weakness and pain, gross edema, gastrointestinal disturbances, liver and kidney damage, swelling of peripheral nerves (neuritis), paralysis
Liver injury jaundice Peripheral vascular disease blackfoot disease Cancer (skin, lung, kidney, bladder)

 Sporadic cases of BFD occurred as early as in the early 20th century, and peak incidence was noted between 1956 and 1960 Human exposure primarily from arsenic contaminated drinking water (seen in southwest coast of Taiwan and Bangladesh) Severe form of peripheral vascular disease in which blood vessels in lower limbs are severely damaged, resulting eventually in progressive gangrene Skin disease:
keratosis of hands and feet, and hyperpigmentation

Blackfoot disease

Decreasing heavy metal toxicity

Minimize/eliminate exposure to sources containing heavy metals
Metallotioneins Phase III efflux transporters Chelating agents

Chelating agents
The word chelation comes from Greek work chele meaning claw Chelators bind directly with metal ions to form stable complexes that remove the metal from competition with the body's cells Because a chelated metal is water soluble, it can be excreted readily by the kidney
For reduction of body burdens associated with toxic metals
DMPS 2,3-dimercapto-1-propanesulfonic acid DMSA dimercaptosuccinic acid D-Penicillamine EDTA ethylenediaminetetraacetic acid

Examples of chelating agents:

1. 2. 3. 4.

 Chelating agents have variable denticity

Denticity refers to the number of atoms in a single ligand (chelator) that bind to a central atom (metal) in a coordination complex Monodentates (1 atom binds to metal) and polydentates (2 or more atoms binds to metal) Most are polydentates (ranging from 2 to 6 atoms) For example, EDTA is a hexadentate ligand while DMPS is a bidentate ligand

DMPS

Desirable attributes for chelating agents

Gain access to the metals Tightly bind and control the metals Not injure recipient Accelerate mobilization and/or removal of the metals Cheap Easy to administer