Veterinary & human medicine

We can help you to better understand the metabolic part of various pathologies

VETERINARY MEDICINE

Veterinarians use clinical chemistry and other laboratory tests to diagnose disease, to monitor disease progression or response to therapy, and to screen for the presence of underlying disease in apparently healthy animals. Clinical chemistry tests are offered by clinical pathology laboratories for this purpose INTEGRACELL is keen to help them by in more specialized topics such as cellular metabolism exploration, oxidative stress and mitochondria dysfunction.

Notion of metabolic disorders in animals

They are of clinical significance because they affect energy production or damage tissues important for survival of the animal. Inherited Metabolic Genetic Disorders do exist in many animals resulting in inborn errors in metabolism. These diseases occur due to absence of enzymes critical in intermediary metabolism. These are progressive in nature and usually are fatal. Examples are diseases associated with decreased RBC survival and anemia such as pyruvate kinase deficiency in Basenjis and Beagles, phosphofructokinase deficiency in English Springer Spaniels. Glucose-6-phosphate dehydrogenase deficiency and hyperkalemic periodic paralysis in horses. α-Mannosidosis in cattle and goat.

Acquired metabolic disorders are primarily related to production or management and metabolism is a critical factor in the pathogenesis of each disease. For example hypoglycemia, promoted by management practices which are aimed at greater production, hence they are entitled production diseases. However, they are also metabolic diseases because the demand for greater production is beyond the capacity of the animal’s metabolic reserves to maintain the particular nutrient at physiologic concentrations.

Ketosis in Cattle and other ruminants

Bovine ketosis is a metabolic disease of lactating dairy cows and occurs worldwide whenever dairy cows are selected and fed for high milk production. Ketosis is basically the result of a negative energy balance in the 6 weeks after parturition. The cow is unable to eat or assimilate enough nutrients to meet her energy needs for maintenance and milk production. Therefore, blood glucose levels drop and body fat and limited protein stores are mobilized in the form of triglycerides and amino acids for gluconeogenesis. Ketone bodies (acetoacetate, β-hydroxybutyrate and acetone) are produced during the mobilization process. Ketone bodies are produced primarily in the liver and in smaller quantities in the mammary gland and rumen wall.

Subclinical ketosis: Postcalving dairy cows with increased BHB, but no clinical signs of are considered to be in a state of subclinical ketosis. Associations have been made between subclinical ketosis and an increased incidence of inflammatory (metritis, mastitis) and metabolic (displaced abomasum, clinical ketosis) diseases postcalving. .
Clinical ketosis: Clinical ketosis typically occurs in cows during early lactation and is most frequently seen in dairy cows Cows with clinical ketosis in dairy herds fed concentrate rations are frequently concurrently hypoglycemic. Blood β-hydroxybutyrate values >27 mg/dL are considered compatible with clinical ketosis. Cows with underlying hepatic lipidosis may have concurrent elevations in liver leakage enzymes or cholestatic enzymes

Pregnancy toxemia in small ruminants: Clinical ketosis due to excess energy demands from the fetus (particulary with twins) also occurs in sheep and goats.

Diabetic ketoacidosis in small animals: Clinical ketosis is seen primarily in small animals as a consequence of diabetes mellitus with a stimulation of gluconeogenesis (which decreases oxaloacetate in hepatocyte mitochondria, facilitating ketogenesis). Affected animals usually have a metabolic acidosis (from accumulation of ketones) and ketonuria. β-hydroxybutyrate concentrations are markedly increased. Rarely, dogs in lactation can also suffer from ketosis. Note that horses have poorly developed ketogenic pathways, so ketosis is rare in this species.

Fatty Liver Disease Of Cattle

Fatty liver is most common in periparturient cattle. It occurs during periods when blood concentrations of non-esterified fatty acids (NEFA) are increased. Oxidation of NEFA leads to the formation of CO2 and ketones, primarily acetoacetate and β-hydroxybutyrate. Fatty liver is often associated with obese cows and downer cows.

Bovine liver diseases

Increase in glutathione peroxidase (GPX) and glucose 6-phosphate dehydrogenase (G6PD) in liver tissue from cattle suffering from liver disease, indicating increased oxidative stress in the liver tissues but not in the blood.

Hypoglycemia

Decreased production of glucose by the liver can occur secondary to inherited defects such as deficiencies of α 1-4 glucosidase (Pompe disease) and glucose-6-phosphatase (von Gierke’s disease). This results in hypoglycemia from defective glycogenolysis.

Juvenile hypoglycemia (usually affects toy and small breed dogs) is due to hepatic immaturity, low liver stores of glycogen and insufficient gluconeogenesis to meet demands. In horses, glucose notably decreases if they are fed a high grain diet, with little roughage.

Hypoglycemia of hunting dogs and endurance horses: A hypoglycemic state is reached from an imbalance in which glucose consumption (glycolysis) occurs at a much faster rate than glucose replenishment (gluconeogenesis and glycogenolysis).

Diabetes Mellitus

In dogs, females are affected twice as often as males. Diabetes mellitus has been associated with bovine virus diarrhea infection in cattle and paramyxovirus infection in llamas, through destruction of pancreatic islets.

Acidosis in Cattle

Acute and chronic acidosis, conditions that follow ingestion of excessive amounts of readily fermented carbohydrate, are prominent production problems for ruminants fed diets rich in concentrate .

Glycolysis: Anaerobic microbes typically thrive when free glucose is available. Rate of glycolysis can be limited by inhibitinghexokinase,, and pyruvate kinase; lack of NAD⁺ also can limit glycolysis.
Volatile Fatty Acid Production and Lactate Production: Bacteria in the rumen often are classified as “lactate producers” or “lactate users. Under anaerobic conditions, pyruvate is converted to lactate to regenerate the NAD used in glycolysis. Conversion of pyruvate to VFA involves multiple steps and generates approximately half the ATP for microbial growth in the rumen; the other half is derived from conversion of glucose to pyruvate.
During sub-acute ruminal acidosis, it has been observed increased values on the concentrations of some acute phase proteins in dairy cows, such as SAA and Haptoglobin

Oxidative stress in animals

There is growing evidence that oxidative stress (OS) significantly impairs organic function and plays a major role in the etiology and pathogenesis of several metabolic diseases in veterinary medicine.

The measurement of hepatic oxidative status in liver biopsy, help in diagnosis of hepatic dysfunction and reflect the degree of deterioration in the liver tissues. On the other hand, an increasing body of evidence suggests that OS is involved in the pathogenesis of a wide range of cardiovascular diseases. The establishment of the specific role of OS in cardiovascular diseases will help to choose the antioxidant therapy that will prove beneficial in combating these problems.

In cardiac diseases in dog, supplementation can play a protective role, avoiding cell disorganization and cellular damages.

Proper anti-oxidant supplementation (coenzyme-Q10, polyphenols, or omega-3 fatty acids) increase the concentration of anti-oxidants in heart cells and make them less sensitive to free radicals.

A number of vitamins and trace minerals are involved in the anti-oxidant defense system and a deficiency of any of these nutrients may depress immunity. Some vitamins (such as Vitamin E or Vitamin C) are important anti-oxidants that have been shown to play an important role in immune-responsiveness and health. A number of trace minerals are required for the functioning of enzymes involved in the anti-oxidant defense system, and certain trace minerals may also affect immune cells via mechanisms distinct from antioxidant properties. Two reports analyze the protective effects of Zinc or Vitamin C in different species (chickens and mice) in different diseases (parasitic infections and haematological disturbances). Finally, OS has been implicated in the pathogenic mechanism of some heavy metals (such as lead or cadmium), causing many disease conditions and toxicities in animals. (Rodriguez et al., 2011)

Our understanding of the role of oxidants and antioxidants in physiological and pathological conditions is continuously increasing and some oxidant-associated or oxidant-mediated processes are now considered as future therapeutic targets. Interestingly, an important part of animal research in this field has been performed in horses, in particular with regard to exercise physiology.

In the page dedicated to “Oxidative Stress in Horses”, authors provide insight into the concept of the oxidant/anti-oxidant equilibrium in horses, by describing how the oxidant/anti-oxidant equilibrium or oxidative stress might be evaluated in the equine species and by presenting current knowledge about oxidative stress in equine medicine.

HUMAN MEDICINE

Cellular metabolism describes a network of biochemical reactions that convert nutrients taken up from the environment into small molecules called metabolites. These metabolites serve as energy equivalents, RedOx co-factors, biomass building blocks, and substrates for DNA/RNA and protein modifications. The metabolism is involved in virtually any cellular process: proliferation and growth signaling, maintenance of ion gradients across membranes, epigenetic remodeling via DNA/protein modifications… Hence, metabolism is highly tissue specific, because it is optimized to the function and cellular processes of the different organs. Moreover, metabolism is tightly interconnected with the upstream signaling network to directly link it with the regulation of dependent cellular processes.

Any of the diseases or disorders that disrupt normal metabolism, the process of converting food to energy on a cellular level, may be considered as a metabolic disease. Metabolic diseases will affect cell ability to perform critical biochemical reactions that involve the processing or transport of proteins (amino acids), carbohydrates (sugars and starches), or lipids (fatty acids). Thousands of enzymes participating in numerous interdependent metabolic pathways carry out this process.

In humans, disruption of normal metabolism is involved in various pathologies, from relatively benign to mostly severe ones.

“Diabetes Mellitus”

It describes a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolisms resulting from defects in insulin secretion, insulin action, or both. The effects of diabetes mellitus include long-term damage, dysfunction and failure of various organs and is a major cause of heart disease and premature death. The number of people with diabetes is rising worldwide. Between 35-40% of people in Europe will develop diabetes over their lifetime.

Two main types of diabetes are described

Type 1 diabetes (TDM1) usually develops in childhood and adolescence and patients require lifelong insulin injections for survival as their pancreas are unable to correctly synthetize then secrete insulin.
Type 2 diabetes (TDM2) usually develops in adulthood and is related to obesity, lack of physical activity, and unhealthy diets. This is the more common type of diabetes (representing 90% of diabetic cases worldwide) and treatment may involve lifestyle changes and weight loss alone, or oral medications or even insulin injections.

In the short term, hyperglycemia causes symptoms of increased thirst, increased urination, increased hunger, and weight loss. However, in the long-term, it causes damage to eyes (leading to blindness), kidneys (leading to renal failure), and nerves (leading to impotence and foot disorders/ possibly amputation). As well, it increases the risk of heart disease, stroke, and insufficiency in blood flow to legs. Many long-term studies have shown that a good metabolic control prevents or delays these complications.

Insuline-resistance and diabetes

Insulin-mediated glucose disposal varies widely in apparently healthy humans, and the more insulin resistant an individual, the more insulin he must secrete in order to prevent the development of type 2 diabetes. However, the combination of insulin resistance and compensatory hyperinsulinemia increases the likelihood that an individual will be hypertensive, and have a dyslipidemia characterized by a high plasma triglyceride (TG) and low high-density lipoprotein cholesterol (HDL-C) concentration. These changes largely increase the risks of cardiovascular disease (CVD).

Metabolic syndrome (MetS)

Metabolic syndrome is a disorder of energy utilization and storage, diagnosed by a co-occurrence of three out of five of the following medical conditions: abdominal (central) obesity, elevated blood pressure, elevated fasting plasma glucose, high serum triglycerides, and low HDL levels. Its pathophysiology appears very complex and has been only partially elucidated. Most patients are elder, obese, sedentary, and have a high degree of insulin resistance. Stress can also be a contributing factor. The most important factors are genetics, aging, diet (particularly sugar-sweetened beverage consumption), sedentary behavior, low physical activity, disrupted chronobiology/sleep, mood disorders/psychotropic medication use, and excessive alcohol use

Its prevalence in elderly population varied from 11% to 43% (median 21%) according to the WHO. Obesity and hypertension are the most prevalent individual components.

MetS and CVD

MetS in elderly is a proven risk factor for cardiovascular morbidity, especially stroke and coronary heart disease (CHD), and mortality. Preventing and treating MetS would be useful in preventing disability and promoting normal aging.

MetS and neurodegenerative disorders

A growing body of epidemiological evidence suggested that MetS and Mets components (impaired glucose tolerance, abdominal or central obesity, hypertension, hypertriglyceridemia, and reduced high-density lipoprotein cholesterol) may be important in the development of age-related cognitive decline (ARCD), mild cognitive impairment (MCI), vascular dementia, and Alzheimer’s disease (AD). It may influence amyloid-beta (Abeta) peptide metabolism and tau protein hyperphosphorylation, the principal neuropathological hallmarks of AD. In AD, an age-related desynchronization of biological systems results, involving stress components, cortisol and noradrenaline, reactive oxygen species, and membrane damage as major candidates that precipitates an insulin resistant brain state (IRBS) with decreased glucose/energy metabolism and the increased formation of hyperphosphorylated tau protein and Abeta.

Cardiovascular diseases

Advanced age is associated with a disproportionate prevalence of CVD. Intrinsic alterations in the heart and the vasculature occurring over the life course render the cardiovascular system more vulnerable to various stressors in late life, ultimately favoring the development of CVD.

The biology of aging has not been fully clarified, but the free radical theory of aging is one of the strongest aging theories proposed to date. The free radical theory has been expanded to the oxidative stress theory, in which mitochondria play a central role in the development of the aging process because of their critical roles in bioenergetics, oxidant production, and regulation of cell death.

Mitochondrial dysfunction is nowadays considered as a major contributor to cardiovascular senescence. Besides being less bioenergetically efficient, damaged mitochondria also produce increased amounts of reactive oxygen species, with detrimental structural and functional consequences for the cardiovascular system. The age-related accumulation of dysfunctional mitochondrial likely results from the combination of impaired clearance of damaged organelles by autophagy and inadequate replenishment of the cellular mitochondrial pool by mitochondriogenesis. A decline in cardiac mitochondrial function associated with the accumulation of oxidative damage might be responsible, at least in part, for the decline in cardiac performance with age.

Lifelong caloric restriction can attenuate functional decline with age, delay the onset of morbidity, and extend lifespan in various species. The effect of caloric restriction appears to be related to a reduction in cellular damage induced by reactive oxygen species.

Cancer

Clinical and epidemiological studies have linked cancer and other chronic medical conditions. For example, patients diagnosed with metS, inflammatory diseases, and autoimmune conditions show increased incidence and aggressiveness of tumor formation. Conversely, diabetics treated with metformin to lower insulin levels have reduced levels of cancer in comparison to untreated individuals. Smoking is linked not only to lung cancer, but also to cardiovascular and other diseases. In general, the molecular bases of these links among diseases are poorly understood.

Inflammation is commonly associated with cancer formation and progression, and it is estimated that 15%–20% of all cancer related deaths can be attributed to inflammation and underlying infections. Inflammatory molecules are elevated in many forms of cancer, and they provide growth signals that promote the proliferation of malignant cells. Constitutively active NF-kB, the key transcription factor that mediates the inflammatory response, occurs in many types of cancer, and mouse models provide evidence for a causative role of NF-kB in malignant conversion and progression.

Metabolism generates oxygen radicals, which contribute to oncogenic mutations. Activated oncogenes and loss of tumor suppressors in turn alter metabolism and induce aerobic glycolysis. Aerobic glycolysis or the Warburg effect links the high rate of glucose fermentation to cancer. Together with glutamine, glucose via glycolysis provides the carbon skeletons, NADPH, and ATP to build new cancer cells, which persist in hypoxia that in turn rewires metabolic pathways for cell growth and survival.

Excessive caloric intake is associated with an increased risk for cancers, while caloric restriction is protective, perhaps through clearance of mitochondria or mitophagy, thereby reducing oxidative stress. Hence, the links between metabolism and cancer are multifaceted, spanning from the low incidence of cancer in large mammals with low specific metabolic rates to altered cancer cell metabolism resulting from mutated enzymes or cancer genes.

Increased cancer risk is associated with obesity, TDM2, high cholesterol, and atherosclerosis, which are components of MetS. Mechanistically, the link between metabolic diseases and cancer is less understood than the connection to inflammation. However, a pathway consisting of AMP-activated protein kinase, an fundamental energy sensor, Akt, and PI3 kinase that are crucial for tumor development plays a critical role in diabetes and other metabolic diseases. In addition, fatty acid synthase also plays an important role in cancer pathogenesis, and inhibitors against this enzyme are being tested as anti-cancer drugs.

Neurodegeneration

Neurodegenerative diseases (NDDs) are traditionally defined as disorders with selective loss of neurons and distinct involvement of functional systems defining clinical presentation.

Most genetic causes of neurodegenerative disorders in childhood are due to neurometabolic disease. Actually, over 200 disorders are described, including aminoacidopathies, creatine disorders, mitochondrial cytopathies, peroxisomal disorders and lysosomal storage disorders. Nevertheless, metabolism dysfunction can not only affect childs but also be a starter for neurodegenerescence in adults that becomes a primary health problem. In 2013, 5.2 million Americans are estimated to be living with Alzheimer’s disease. By 2050, the number of people age 65 and older with the disease is projected to be nearly 14 million. An estimated 1 million Americans currently live with Parkinson’s disease, and the prevalence of the disease is expected to increase substantially in the next 20 years due to the aging of the population.

The human brain has the highest energy demands of any organ in the body, consuming more than 20% of the body’s glucose and oxygen, despite comprising only 2% of total body mass. The majority of this energy is used to support functional processes. Numerous studies have reported that metabolic rates of glucose and oxygen decline with age, and in an accelerated manner in Alzheimer senile dementia (AD). The metabolic rate reductions have been considered as major contributors to brain structural alteration (gray matter and white matter atrophy) and cognitive impairment later in life.

CNS functions strongly depend on efficient mitochondrial function, because brain tissue has a high energy demand. Mutations in the mitochondrial genome, defects in mitochondrial dynamics, generation and presence of ROS, protein aggregate-associated dysfunctions and environmental factors may alter energy metabolism and in many cases are associated with neurodegenerative diseases.

Aside direct involvement of mitochondrial dysfunction, accumulating evidence indicates that insulin plays an important role in the regulation of brain glucose homeostasis in the central nervous system and has trophic effects on neurons. Thus, insulin metabolism dysfunction in diabetes may contribute directly to AD, likely via the accelerated formation of advanced glycation end products (AGEs) in the brain. Defective insulin secretion and insulin resistance in the tissues means too much glucose in the blood and also in the brain. This presence can lead to an increased oxidative stress. AGEs, along with other signs of oxidative damage, have been found lurking inside the pathology of many other adult-onset neurodegenerative disorders, including Pick’s disease, Parkinson’s, Progressive Supranuclear Palsy, Lewy Body Dementia, and ALS. Each of these involves a progressive decline in neurological function with deficits that may include dementia, movement disorders, seizures, muscle weakness…, that can be tightly associated with pathological expression of protein aggregates that can be seen microscopically in the nervous tissue. Brain accumulation of senile Aβ plaques and hyperphosphorylated tau (neurofibrillary tangles) in the medial temporal lobe (MTL) and cortical areas of the brain of a patient with mild cognitive impairment is associated with a high risk of developing AD. This precipitation of modified protein forming aggregates can be related to the protein side-chains modifications either directly by reactive oxygen species (ROS) or reactive nitrogen species (RNS), or indirectly, by the products of lipid peroxidation.