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Shilajit attenuates behavioral symptoms of chronic fatigue syndrome by modulating the hypothalamic–pituitary–adrenal axis and mitochondrial bioenergetics in rats
Abstract
Shilajit has been used as a rejuvenator for ages in Indian ancient traditional medicine and has been validated for a number of pharmacological activities.
The effect of processed shilajit which was standardized to dibenzo-α-pyrones (DBPs;0.43% w/w), DBP-chromoproteins (DCPs; 20.45% w/w) and fulvic acids (56.75% w/w) was evaluated in a rat model of chronic fatigue syndrome (CFS). The mitochondrial bioenergetics and the activity of hypothalamus-pituitary-adrenal (HPA) axis were evaluated for the plausible mechanism of action of shilajit.
CFS was induced by forcing the rats to swim for 15mins for 21 consecutive days. The rats were treated with shilajit (25, 50 and 100mg/kg) for 21 days before exposure to stress procedure. The behavioral consequence of CFS was measured in terms of immobility and the climbing period. The post-CFS anxiety level was assessed by elevated plus maze (EPM) test. Plasma corticosterone and adrenal gland weight were estimated as indices of HPA axis activity. Analysis of mitochondrial complex chain enzymes (Complex I, II, IV and V) and mitochondrial membrane potential (MMP) in prefrontal cortex (PFC) were performed to evaluate the mitochondrial bioenergetics and integrity respectively.
Shilajit reversed the CFS-induced increase in immobility period and decrease in climbing behavior as well as attenuated anxiety in the EPM test. Shilajit reversed CFS-induced decrease in plasma corticosterone level and loss of adrenal gland weight indicating modulation of HPA axis. Shilajit prevented CFS-induced mitochondrial dysfunction by stabilizing the complex enzyme activities and the loss of MMP. Shilajit reversed CFS-induced mitochondrial oxidative stress in terms of NO concentration and, LPO, SOD and catalase activities.
The results indicate that shilajit mitigates the effects of CFS in this model possibly through the modulation of HPA axis and preservation of mitochondrial function and integrity. The reversal of CFS-induced behavioral symptoms and mitochondrial bioenergetics by shilajit indicates mitochondria as a potential target for treatment of CFS.
... Mitochondrial function can be compromised by a multitude of factors, including reduced availability of necessary metabolites and mitochondrial damage through, for example, oxidative stress (Box 1). The notion that mitochondrial dysfunction can lead to fatigue is indicated by the observation that fatigue is a common symptom of mitochondrial disease (Gorman et al., 2015) and that fatigue-like behavior in animal models is associated with reduced expression of mitochondrial complexes, reductions in metabolic activity, and alterations in mitochondrial morphology in the brain (Surapaneni et al., 2012;Wang et al., 2014;Vichaya et al., 2016). ...
... All groups showed reductions in brain MDA. Surapaneni et al., 2012 Forced swim (15 min/day for 21 days) as a model of CFS in rats Evaluated behavior and measures of mitochondria function in control mice and those supplemented during the 21 days with withania somnifera and shilajit. ...
Chronic or persistent fatigue is a common, debilitating symptom of several diseases. Persistent fatigue has been associated with low-grade inflammation in several models of fatigue, including cancer-related fatigue and chronic fatigue syndrome. However, it is unclear how low-grade inflammation leads to the experience of fatigue. We here propose a model of an imbalance in energy availability and energy expenditure as a consequence of low-grade inflammation. In this narrative review, we discuss how chronic low-grade inflammation can lead to reduced cellular-energy availability. Low-grade inflammation induces a metabolic switch from energy-efficient oxidative phosphorylation to fast-acting, but less efficient, aerobic glycolytic energy production; increases reactive oxygen species; and reduces insulin sensitivity. These effects result in reduced glucose availability and, thereby, reduced cellular energy. In addition, emerging evidence suggests that chronic low-grade inflammation is associated with increased willingness to exert effort under specific circumstances. Circadian-rhythm changes and sleep disturbances might mediate the effects of inflammation on cellular-energy availability and non-adaptive energy expenditure. In the second part of the review, we present evidence for these metabolic pathways in models of persistent fatigue, focusing on chronic fatigue syndrome and cancer-related fatigue. Most evidence for reduced cellular-energy availability in relation to fatigue comes from studies on chronic fatigue syndrome. While the mechanistic evidence from the cancer-related fatigue literature is still limited, the sparse results point to reduced cellular-energy availability as well. There is also mounting evidence that behavioral-energy expenditure exceeds the reduced cellular-energy availability in patients with persistent fatigue. This suggests that an inability to adjust energy expenditure to available resources might be one mechanism underlying persistent fatigue.
... The effects of a processed and standardized shilajit (25, 50, and 100 mg/kg/day for 21 days) on various stress factors in rats forced to swim 15 min per day for 21 days were assessed (Surapaneni et al., 2012). The product contained 0.43% DBPs, 20.45% DPB-chromoproteins, and 56.75% fulvic acids. ...
... Antioxidant status was assessed based on glutathione peroxidase activity, glutathione reductase activity, catalase activity, glutathione content, and lipid peroxidation (thiobarbituric acid reactive substances). Shilajit treatment (25, 50, and 100 mg/kg/day for 21 days) also attenuated swimming-induced oxidative stress as evidenced by decreases in nitric oxide and lipid peroxidation and increases in catalase and superoxide dismutase activities (Surapaneni et al., 2012). ...
Shilajit (mumie, moomiyo, mummiyo, mumijo, salajeet) is a resinous phyto-mineral exudate found in sedimentary rocks that has been used for many years for a wide variety of illnesses and health benefits. In recent years, a growing number of studies have been published involving humans, animal, and in vitro systems in support of its uses and health-related effects. Animal and human studies support its use as a “revitalizer,” promoting physical and mental energy, enhancing physical performance, and relieving fatigue in association with enhanced ATP production. Various published research studies indicate that shilajit exhibits adaptogenic, antioxidant, antiinflammatory, immunomodulatory, antidiabetic, and neurological properties. Studies also show that shilajit enhances spermatogenesis. Based on animal and human studies, the safety of shilajit is well documented. Shilajit is standardized to fulvic acids, and key constituents in shilajit responsible for its effects appear to be fulvic acids comprising oxygenated dibenzo-a-pyrones (DBPs) and their derivatives. The current literature regarding the efficacy and safety of shilajit is reviewed.
... The elevated intracellular reactive oxygen species aggravate exercise induced muscular weakness and fatigue. 2 Further a strenuous exercise may induce mitochondrial dysfunction which cause oxidative damage and injury to tissues. [3][4][5][6] Myostatin a transforming growth factor-β superfamily plays vital role in regulation of skeletal muscle mass. Rigorous exercise modulates Myostatin mRNA expression in skeletal muscles affecting the muscle mass. ...
... Further both MIMBP and PMIMBP prevented the attenuation of mitochondrial SDH activity. The improved activity of M a n u s c r i p t complex I and II suggests increased rate of transfer of electrons into the ETC.5 Cytochrome C catalyzes electron transport from ubiquinone to cytochrome oxidase. ...
Background: The current study was designed to investigate the influence of monofloral Indian mustard bee pollen (MIMBP) and processed monofloral Indian mustard bee pollen (PMIMBP) supplementation on chronic swimming exercise-induced oxidative stress implications in the gastrocnemius muscle of Wistar rats.
Methods: MIMBP was processed with an edible lipid-surfactant mixture (Captex 355:Tween 80) to increase the extraction of polyphenols and flavonoid aglycones as analyzed by UV spectroscopy and high performance liquid chromatography-photo diode array. Wistar rats in different groups were fed with MIMBP or PMIMBP supplements at a dose of 100 mg/kg, 200 mg/kg and 300 mg/kg individually, while being subjected to chronic swimming exercise for 4 weeks (5 d/wk). Various biochemical [superoxide dismutase (SOD), glutathione (GSH), malonaldehyde (MDA), nitric oxide (NO), and total protein content], mitochondrial (Complex I, II, III, and IV enzyme activity), and molecular (myostatin mRNA expression) parameters were monitored in the gastrocnemius muscle of each group.
Results: Administration of both MIMBP (300 mg/kg) and PMIMBP (100 mg/kg, 200 mg/kg, and 300 mg/kg) wielded an antioxidant effect by significantly improving SOD, GSH, MDA, NO, and total protein levels. Further MIMBP (300 mg/kg) and PMIMBP (200 mg/kg and 300 mg/kg) significantly improved impaired mitochondrial Complex I, II, III, and IV enzyme activity. Significant down-regulation of myostatin mRNA expression by MIMBP (300 mg/kg) and PMIMBP (200 mg/kg and 300 mg/kg) indicates a muscle protectant role in oxidative stress conditions.
Conclusion: The study establishes the antioxidant, mitochondrial upregulatory, and myostatin inhibitory effects of both MIMBP and PMIMBP in exercise-induced oxidative stress conditions, suggesting their usefulness in effective management of exercise-induced muscular stress. Further, processing of MIMBP with an edible lipid-surfactant mixture was found to improve the therapeutic efficiency of pollen.
... We are still a long way from understanding the evolutionary advantage of the altered regulation of the HPA axis in patients who have, and those who go on to develop, CFS; perhaps CFS is the cost of the cortisol response to challenge (including social challenge) which may be necessary to adapt to the complex dynamics of human social competition [71]. Another potential cause of disruption to HPA axis function is oxidative stress and a decrease in antioxidant capacity in addition to the presence of histone deacetylase (HDAC) [72]. Increased activity of HDAC 2 and 3 coincides with a decrease in plasma cortisol [14]. ...
... There is an argument for further trials of steroid treatment in patients selected on the basis of adrenal insufficiency, but the potential impact of long-term treatment including Cushing's syndrome, osteoporosis , extreme mood changes, and seizures cautions against this approach [99]. The HPA axis though remains a potential target for novel treatment strategies in CFS and this has been examined in studies utilizing animal models to examine traditional medicines with a putative HPA axis effect; for example, Shilajit, a traditional Indian medicine, reduced immobility and increased climbing behavior whilst increasing adrenal weight and corticosterone levels in the forced swim test in rats [72] and Myelophil, based on compounds used for fatigue in Chinese medicine, increased glucocorticoid receptor expression in the hypothalamus and hippocampus , and altered expression of cytokines such as interleukin (IL-10) and tumour necrosis factor-alpha (TNF-í µí»¼) using the chronic cold stress and restraint model in mice [100]. One of the most interesting proposals is the switch to a new steady state from chronic hypocortisolaemia to a healthy, reactive state using the model-based predictive control (MPC) solution originally proposed by Gupta and colleagues [35]. ...
Hypothalamic-pituitary-adrenal (HPA) axis dysfunction has been found in a high proportion of chronic fatigue syndrome (CFS) patients and includes enhanced corticosteroid-induced negative feedback, basal hypocortisolism, attenuated diurnal variation, and a reduced responsivity to challenge. A putative causal role for genetic profile, childhood trauma, and oxidative stress has been considered. In addition, the impact of gender is demonstrated by the increased frequency of HPA axis dysregulation in females. Despite the temporal relationship, it is not yet established whether the endocrine dysregulation is causal, consequent, or an epiphenomenon of the disorder. Nonetheless, given the interindividual variation in the effectiveness of existing biological and psychological treatments, the need for novel treatment strategies such as those which target the HPA axis is clear.
... COX plays a central role in energy metabolism and is reported to protect the aging nerve cells from damage, but with aging, its low expression becomes the reason for neurodegeneration [68] . Age-related deterioration in the COX function was linked with corresponding variations in mitochondrial membrane potential, respiration and an upsurge July-August 2023 Indian Journal of Pharmaceutical Sciences attenuated oxidative stress in the hypothalamic paraventricular nucleus of spontaneously hypertension rats [82,83] . ...
Phytoconstituents epigallocatechin gallate and withaferin A, found in Camellia sinensis (Kangra green tea) and Withania sominifera (Ashwagandha) respectively, were explored for their binding affinity towards various enzymes involved in the skin-aging process. Epigallocatechin gallate and withaferin A were analyzed for their physiochemical properties, drug-likeness and human intestinal absorptivity using Data Warrior, Molsoft and SwissADME (boiled egg model) respectively. Molecular docking analysis for different enzymes involved in aging (collagenase, elastase and hyaluronidase), antioxidant enzymes (superoxide dismutase, glutathione-s-transferase, glutathione peroxidase and catalase) and mitochondrial enzymes (nicotinamide adenine dinucleotide (NAD)+hydrogen (H) dehydrogenase, succinate dehydrogenase, cytochrome c oxidase and adenosine triphosphate synthase) was carried out for epigallocatechin gallate alone (1), withaferin A alone (2), epigallocatechin gallate and withaferin A in combination (3) and a reference molecule. Autodock Vina was employed to carry out individual molecular docking as well as multiple ligand simultaneous docking. The results were analyzed in terms of binding energy and different interacting residues. Interestingly, (3) displayed a higher binding affinity towards all the aging and antioxidant enzymes as compared to (1), (2) and the references. Moreover, the combination of the constituents exhibited better binding for most of the mitochondrial enzymes. Additionally, molecular dynamics simulations were performed to estimate stability and flexibility of best complexes, while collagenase activity colorimetric assay was carried out to study the effects of (1), (2) and (3) on collagenase. The in vitro analysis indicated a 1.5 times increase in collagenase inhibition upon using (3) as compared to ascorbic acid (standard). Overall, the results indicate that epigallocatechin gallate and withaferin A, in combination, may potentially inhibit skin-aging, while enhancing antioxidant effects of various enzymes, and warrant further experimental validation.
... Shilajit enhances CoQ10's mitochondrial benefits and supports levels of the active ubiquinol form. Components of shilajit can serve as electron reservoirs, replenishing electrons lost by CoQ10 and allowing this vital coenzyme to remain active longer (Surapaneni, 2012). ...
... Other anticipated pathways contributing to Antioxidant properties of Shilajatu • Studies have reported that Shilajatu significantly increases the SOD (Superoxide dismutases) and Catalase activity. 31 SODs are a group of metalloenzymes that are found in all kingdoms of life. SODs act as the front line of defence against ROS mediated injury. ...
... In a study on rat model it has been observed that the shilajatu attenuate the behavioural symptoms of chronic fatigue syndrome possibly through the modulation of hypothalamus-pituitary-adrenal (HPA) axis and preservation of mitochondrial function and integrity [55] . In an experimental study on effect of shilajatu administration on experimentally induced osteoarthritis in rats, it has been observed that the group of rats which has been treated with shilajatu aqueous extract, has less cartilage degenerative changes than the group which has not been administered with the same [56] . In a study on dogs, researchers have concluded that Shilajatu has significant anti-arthritic properties including reduction of pain and inflammation [57] . ...
... One of the famous Traditional Chinese Medicines used to treat CFS, Herba Epimedii, can up-regulate the expression of noradrenaline to improve HPA-axis hypoactivity of CFS [119]. A rejuvenator for ages in ancient Indian traditional medicine, shilajit, mitigates the effects of CFS possibly through the modulation of HPA axis and preservation of mitochondrial function and integrity [120]. ...
It is a common phenomenon that people are in a sub-health condition and facing “unexplained fatigue”, which seriously affects their health, work efficiency and quality of life. Meanwhile, fatigue is also a common symptom of many serious diseases such as HIV/AIDS, cancer, and schizophrenia. However, there are still no official recommendations for the treatment of various forms of fatigue. Some traditional natural medicines are often used as health care products, such as ginseng, Cordyceps militaris (L.ex Fr.Link) and Rhodiola rosea L., and these have been reported to have specific anti-fatigue effects with small toxic and side effects and rich pharmacological activities. It may be promising treatment strategy for sub-health. In this review, we first outline the generation of fatigue. Furthermore, we put emphasis on the anti-fatigue mechanism, bioactive components, and clinic trials of natural medicines, which will contribute to the development of potential anti-fatigue agents and open up novel treatments for sub-health.
... Shilajit enhances CoQ10's mitochondrial benefits and supports levels of the active ubiquinol form. Components of shilajit serve as electron reservoirs, replenishing electrons lost by CoQ10 and allowing this vital coenzyme to remain active longer (93). ...
... Shilajit enhances CoQ10's mitochondrial benefits and supports levels of the active ubiquinol form. Components of shilajit serve as electron reservoirs, replenishing electrons lost by CoQ10 and allowing this vital coenzyme to remain active longer (93). ...
... Shilajit enhances CoQ10's mitochondrial benefits and supports levels of the active ubiquinol form. Components of shilajit can serve as electron reservoirs, replenishing electrons lost by CoQ10 and allowing this vital coenzyme to remain active longer (Surapaneni, 2012). ...
Mitochondrial dysfunction has been identified as one of the principal causes of bioenergetic decline. Although there is no single silver bullet or an exact combination of substances or supplements that will unfailingly resuscitate all aspects of failing mitochondria, it has been reported that a number of nutrients, supplements and prescription substances may alleviate or restore many aspects of mitochondrial failure.
Combinations of these, acting on multiple targets, may normalize and/or improve mitochondrial function, increase cellular and systemic energy production, alleviate mitochondrial- related disease, and delay age-related decline in many
organs and systems of the body. The rise in the incidence of cancer and deaths from cancer not only parallels the rise in the development and use of toxic chemicals and materials in the environment, but also toxins in our food and water supplies and pharmaceuticals. The rise in cancer incidence and deaths is thought to be directly caused by such toxic ingestion and the body’s increasing inability to cope with the toxic overload of xenobiotics that profoundly affects the mitochondria. It is conceivable that combinations of various mitochondrial enhancers/resuscitators, acting on various portions of the mitochondrial energy production pathway will have complementary/additive effects and decrease the cancer incidence and death rates. Here we have proposed a combination of diet, exercise and supplements containing a mixture of nutrients mentioned herein to significantly enhance mitochondrial function to help restore oxidative respiration to a level of favoring malignant cell re-differentiation or to at least restore apoptotic mechanisms since the intrinsic apoptotic pathway in cells is regulated largely by functional mitochondria. When restoring mitochondrial function, we may reverse aerobic glycolysis,
inhibit cancer cell growth and possibly, reverse malignancy. Scientific support for the use of vitamin-based and cofactor-based mitochondrial therapies is accumulating. This
Mitochondrial Correction (Mitochondrial Rescue, Mitochondrial repair) approach is intended to promote critical enzymatic reactions, reduce putative sequelae of excess free
radicals, and scavenge toxic metabolic molecules, which tend to accumulate in mitochondrial diseases. Some supplements also may act as alternative energy fuels or may bypass biochemical blocks within the respiratory chain. We believe this concept can have an important repercussion in the treatment of degenerative diseases.
... Further eugenol was evaluated against restraint stress-induced gastrointestinal dysfunction (Garabaduet al., 2015). Shilajit attenuated behavioural symptoms of chronic fatigue syndrome by modulating the hypothalamicpituitary-adrenal axis and mitochondrial bioenergetics in rats (Surapaneni et al., 2012). Asparagus racemosus modulated the hypothalamic-pituitaryadrenal axis and brain monoaminergic systems in rats (Krishnamurthy et al., 2013a). ...
Neurological and neuropsychiatric problems have been recognized as formidable issue worldwide in developed as well as developing countries due to a sizable mortality and debilitating sequelae. A lot of impetus on research in this area over the past two decades has led to a better understanding of these diseases and made available tools in the form of experimental model systems, mechanistic pathways and genetic connections to provide the researchers with an enabling environment for drug discovery, better value from existing drugs and circumventing toxic effects. In India, the past decade has witnessed a broadening of research base of neuropharmacology research and a close look reveals that Indian research is very much contemporary. A kaleidoscopic view of neuropharmacology research in India in over the past five years is presented in this article.
... It contains flavonoids and polyphenols which are natural antioxidants and significantly increases glutathione, catalase activities, and superoxide dismutase, which are the first line defensive enzymes against free radicals. [52][53][54]Lauha bhasma reduces the degree of oxidative stress signaling pathways and by that preventing insulin resistance and β-cells dysfunction and ultimately controlling the blood sugar level. [55] Medohara (hypolipidemic) effect of Lauha bhasma decreases the high lipid level. ...
Objectives: To assess the antidiabetic potential of Shilajatvadi Lauha (SL) processed with Daruharidra (modified SL [MSL]) in streptozotocin (STZ)-nicotinamide (NA)-induced diabetic rats.
Materials and Methods: Animals were divided into diabetic and nondiabetic groups. Type 2 diabetes in rats was induced with a single dose of STZ (65 mg/kg) NA (110 mg/kg) intraperitoneal diabetic rats were treated with formulation MSL (10, 30, and 100 mg/kg) and glibenclamide (10 mg/kg) once daily for 14 days orally. After 14 days treatment, fasting blood glucose and plasma insulin were assayed. Different biochemical parameters such as total cholesterol (TC), triglycerides (TGs), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), and very LDL-C (VLDL-C) were also examined.
Results: MSL significantly lowered the blood glucose and increases insulin level, which was comparable to the standard antidiabetic drug, glibenclamide. Treatment with MSL showed a significantly reduction in the levels of TC, TG, LDL-C, and VLDL-C and increases the level of HDL-C.
Conclusion: MSL possess significant antidiabetic and antihyperlipidemic activity in Type 2 diabetes mellitus rats. The results are encouraging and further studies can be used to evaluate the exact mechanism of action to develop a novel molecule that will benefit the ailing.
... It contains flavonoids and polyphenols which are natural antioxidants and significantly increases glutathione, catalase activities, and superoxide dismutase, which are the first line defensive enzymes against free radicals. [52][53][54] Lauha bhasma reduces the degree of oxidative stress signaling pathways and by that preventing insulin resistance and β-cells dysfunction and ultimately controlling the blood sugar level. [55] Medohara (hypolipidemic) effect of Lauha bhasma decreases the high lipid level. ...
Objectives: To assess the antidiabetic potential of Shilajatvadi Lauha (SL) processed with
Daruharidra (modified SL [MSL]) in streptozotocin (STZ)‑nicotinamide (NA)‑induced
diabetic rats.
Materials and Methods: Animals were divided into diabetic and nondiabetic groups.
Type 2 diabetes in rats was induced with a single dose of STZ (65 mg/kg) NA (110 mg/kg)
intraperitoneal diabetic rats were treated with formulation MSL (10, 30, and 100 mg/kg)
and glibenclamide (10 mg/kg) once daily for 14 days orally. After 14 days treatment, fasting
blood glucose and plasma insulin were assayed. Different biochemical parameters such as
total cholesterol (TC), triglycerides (TGs), low‑density lipoprotein‑cholesterol (LDL‑C),
high‑density lipoprotein‑cholesterol (HDL‑C), and very LDL‑C (VLDL‑C) were also examined.
Results: MSL significantly lowered the blood glucose and increases insulin level, which was
comparable to the standard antidiabetic drug, glibenclamide. Treatment with MSL showed
a significantly reduction in the levels of TC, TG, LDL‑C, and VLDL‑C and increases the
level of HDL‑C.
Conclusion: MSL possess significant antidiabetic and antihyperlipidemic activity in Type 2
diabetes mellitus rats. The results are encouraging and further studies can be used to evaluate
the exact mechanism of action to develop a novel molecule that will benefit the ailing.
... The effectiveness of Shilajit pharmaceutical formulations on wound healing surely has been not proven in clinical studies in animals [11]. May confirmations of some findings have come from animal (mainly rats or mice) and in vitro studies [12]. Vishnevsky ointment contains essential agents such as birch tar, xeroformium and castor oil which are used for management of wounds, burns, skin ulcer and suppurations [13]. ...
Introduction: Renal cell carcinoma (RCC) is known to be chemo resistant but with the introduction of targeted therapies; there has been a “revolution” in its treatment strategies. The only targeted therapy available in Tunisia for the treatment of metastatic and/or locally advanced RCC is sunitinib. Objective of the Study: To evaluate therapeutic results and tolerance of sunitinib in metastatic and/or locally advanced RCC. Subjects and Methods: This was a retrospective study covering a period of six years (from January 2008 to January 2014) conducted in 5 medical oncology departments in Tunisia. The population of the study consisted of 29 patients treated with sunitinib for metastatic and/or locally advanced RCC. Results: The mean age of patients was 51 years. Three patients had tumor recurrence and 26 patients had a metastatic RCC. The prognosis was good for 5 patients, intermediate for 19 patients and poor for 5 patients. The median duration of treatment was 5 months. Because of side effects, treatment was discontinued in 12.5% of cases and the dose was reduced in 10.3% of cases. Side effects consisted of asthenia (95.8%), stomatitis (70.8%), anemia (50%), hand-foot syndrome (55.8%) in addition to nausea and vomiting (54.2%). Objective response was observed in 37.5% of patients after 3 months of treatment and in 50% after 6 months. The median progression-free survival was 14 months (95% CI, 7.9 to 20.6). The median overall survival was 22 months (95% CI, 15.6 to 28.7). Conclusion: The prognosis of RCC in Tunisian patients has clearly improved with the introduction of sunitinib, but other therapies with a proven efficacy as a first and second line therapy should be considered.
... The effectiveness of Shilajit pharmaceutical formulations on wound healing surely has been not proven in clinical studies in animals [11]. May confirmations of some findings have come from animal (mainly rats or mice) and in vitro studies [12]. Vishnevsky ointment contains essential agents such as birch tar, xeroformium and castor oil which are used for management of wounds, burns, skin ulcer and suppurations [13]. ...
To compare the healing of purulent wounds with Shilajit dressing vs. Vishnevsky dressing in dogs with purulent wounds of >2 weeks of duration, eight dogs were chosen from same ages in two groups i.e., Shilajit and Vishnevsky dressing group. Dressing was done on every day basis for more than two weeks of follow up period. Main outcome of healing measure was completed at three weeks. Wound healing status was assessed at three days intervals till end of three weeks. Shilajit treated achieved complete decrease in the wound surface area, effect of healing score in Shilajit dressing group in comparison to the Vishnevsky dressing group at p > 0.05 level of significance. Shilajit is highly effective in achieving a characteristic feature of regenerative and granulation healing of purulent wounds as compared to Vishnevsky dressing group in dogs.
... The protective effects of this substance may be due to antioxidant activities in shilajit (especially fulvic acid), protecting the body from cellular damages (22,23). Shilajit functions via improving the activities of mitochondria at cellular level, subsequently increasing the levels of energy (24). The increase and maintenance of cellular energy levels can save the tissues from the deterioration. ...
Asphaltum (Shilajit) is a blackish-brown exudation available in different consistency which deposits on the rocks of different mountain ranges especially Hindukash and Himalayas in Indian subcontinent. It is composed of 220 mineral and metal substances used in traditional Indian medical systems. Shilajit provides beneficial effects for physical strengthening, improving urinary tracks functioning and stabilizes blood sugar. It also has immune-modulation effects, increases brain potency, anti-arthritis, anti-aging and anti-hypertension activity. In oriental medicines of Asian countries, Shilajit has also been ascribed as a potent aphrodisiac and used to treat male sexual dysfunction. It has been reported that Shilajit increases serum testosterone level and sperm number in male rat and human. ÖZET Asphaltum (Shilajit), özellikle Hindistan karaparçasındaki Hindikuş ve Himalaya dağlarındaki kayalarda bulunan, değişik kıvamda olabilen, siyahımsı-kahverengi bir maddedir. Asphaltum, geleneksel Hindistan Tıbbı'nda kullanılan 220 kadar mineral ve madde içerir. Shilajit, fiziksel olarak iyi hissetme, üriner sistem fonksiyonlarında iyileşme ve kan şekerinin stabil hale gelmesi gibi faydalı etkilere neden olur. İlave olarak, immün modülasyon, beyin fonksiyonlarında artış, anti-artritik, yaşlanma karşıtı ve antihipertansif etkileri vardır. Shilajit, Asya ülkelerinin geleneksel tıbbında afrodizyak olarak bilinir ve erkek seksüel bozukluklarının tedavisinde kullanılır. Shilajit'in erkek ratlarda ve insanlarda sperm sayısını ve serum testosterone seviyesini arttırdığı rapor edilmiştir.
... However, several lines of evidence have shown significantly reduced HPA axis function in stress-related CFS (Gold andChrousos, 2002, Fries et al., 2005). We suspect that restraint stress and forced swimming act as stressors that activate the HPA axis response, while the contradictory results of other studies may reflect different animal models involving alternative stressors inducing CFS (Surapaneni et al., 2012). Our RT-PCR and ELISA results suggest that moxibustion modulates the stress response via the HPA axis, which is manifested by reduced hypothalamic CRH mRNA expression and plasma ACTH and CORT levels. ...
In China, moxibustion is reported to be useful and has few side effects for chronic fatigue syndrome, but its mechanisms are largely unknown. More recently, the focus has been on the wealth of information supporting stress as a factor in chronic fatigue syndrome, and largely concerns dysregulation in the stress-related hypothalamic-pituitary-adrenal axis. In the present study, we aimed to determine the effect of moxibustion on behavioral symptoms in chronic fatigue syndrome rats and examine possible mechanisms. Rats were subjected to a combination of chronic restraint stress and forced swimming to induce chronic fatigue syndrome. The acupoints Guanyuan (CV4) and Zusanli (ST36, bilateral) were simultaneously administered moxibustion. Untreated chronic fatigue syndrome rats and normal rats were used as controls. Results from the forced swimming test, open field test, tail suspension test, real-time PCR, enzyme-linked immunosorbent assay, and western blot assay showed that moxibustion treatment decreased mRNA expression of corticotropin-releasing hormone in the hypothalamus, and adrenocorticotropic hormone and corticosterone levels in plasma, and markedly increased progranulin mRNA and protein expression in the hippocampus. These findings suggest that moxibustion may relieve the behavioral symptoms of chronic fatigue syndrome, at least in part, by modulating the hypothalamic-pituitary-adrenal axis and upregulating hippocampal progranulin.
... Ingredients like bioactive dibenzo alpha pyrones, along with humic and fulvic acids present in Shilajit, have been reported to induce physiological actions (12). Shilajit has been found to increase the plasma corticosterone levels and to decrease adrenal gland weight during chronic fatigue syndrome, and to prevent mitochondrial dysfunction by stabilizing the complex mitochondrial enzymes (30), and has been reported to show anxiolytic activity in rodents (19). Clinical research has confirmed that some ingredients of Shilajit are quickly absorbed through the intestinal tract and once in the systemic circulation, it can penetrate the bloodbrain barrier (21). ...
Shilajit, a mineral pitch, has been used in Ayurveda and Siddha system of medicine to treat many human ailments, and is reported to contain at least 85 minerals in ionic form. This study examined the possible mechanism of Shilajit action on preoptic hypothalamic neurons using juvenile mice. The hypothalamic neurons are the key regulator of many hormonal systems. In voltage clamp mode at a holding potential of -60 mV, and under a high chloride pipette solution, Shilajit induced dose-dependent inward current. Shilajit-induced inward currents were reproducible and persisted in the presence of 0.5 μM tetrodotoxin (TTX) suggesting a postsynaptic action of Shilajit on hypothalamic neurons. The currents induced by Shilajit were almost completely blocked by 2 μM strychnine (Stry), a glycine receptor antagonist. In addition, Shilajit-induced inward currents were partially blocked by bicuculline. Under a gramicidin-perforated patch clamp mode, Shilajit induced membrane depolarization on juvenile neurons. These results show that Shilajit affects hypothalamic neuronal activities by activating the strychnine-sensitive glycine receptor with α₂/α₂β subunit. Taken together, these results suggest that Shilajit contains some ingredients with possible glycine mimetic activities and might influence hypothalamic neurophysiology through activation of strychnine-sensitive glycine receptor-mediated responses on hypothalamic neurons postsynaptically.
... It contains flavonoids and polyphenols which are natural antioxidants and significantly increases superoxide dismutase, glutathione, and catalase activities which are first-line defensive enzymes against free radicals. [27][28][29][30] Rasayana effect of Lauha bhasma reduces the degree of oxidative stress signaling pathways and, by that, preventing insulin resistance and β-cells dysfunction and, ultimately, controlling the blood sugar level, and its Medohara (hypolipidemic) effect decreases the high lipid level. [31] Copper (Cu) is one of the major constituents of Swarna makshik bhasma which is responsible for cholesterol and glucose metabolism in the body, [8,32] Trikatu; Shunthi (Zingiber officinale Roscoe), Marich (Piper nigrum L), and Pippali (Piper longum L) increase bioavailability by promoting rapid absorption from the gastrointestinal tract, or preventing metabolism/oxidation during the first passage through the liver after being absorbed, or a combination of these mechanisms, helping improve most drugs therapeutic activity, and it possess anti-glycation and antioxidants properties too which reduces the blood sugar level. ...
Context: Shilajatvadi Lauha (SL) is used in Ayurveda as Indian traditional medicine for treating diabetes mellitus. Aims: To explore the anti-diabetic potential of SL in nicotinamide-streptozotocin induced diabetic rats. Materials and Methods: SL (10, 30, and 100 mg/kg) and glibenclamide (10 mg/kg) were orally administered once daily to diabetic rats for 14 days. Blood glucose, plasma insulin, total cholesterol (TC), triglycerides (TGs), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), and very LDL-C (VLDL-C) were examined. Results: SL significantly lowered the blood glucose without any hypoglycemic effect on their control counterparts, which was comparable to that of the standard anti-diabetic drug, glibenclamide. SL also showed reduction in the levels of TC, TGs, LDL-C, VLDL-C, but it increases the levels of plasma insulin and HDL-C in diabetic rats. Conclusions: SL possesses anti-diabetic and anti-hyperlipidemic activities in Type 2DM rats, which seems to scientifically validate its traditional uses and might be a promising drug in the therapy of diabetes mellitus and its hyperlipidemic complications.
... In another paper, BALBc mice were injected with Brucella abortus and developed decreased running activity that lasted one week [57]. Forced swim of Charles Foster albino rats for twenty-one days also increased immobility time, anxiety as assessed by elevated plus maze test, and brain oxidative stress [58]. These findings maybe due to the differences in the strains and triggers used. ...
Background
Chronic Fatigue Syndrome (CFS) is a neuroimmunoendocrine disease affecting about 1% of the US population, mostly women. It is characterized by debilitating fatigue for six or more months in the absence of cancer or other systemic diseases. Many CFS patients also have fibromyalgia and skin hypersensitivity that worsen with stress. Corticotropin-releasing hormone (CRH) and neurotensin (NT), secreted under stress, activate mast cells (MC) necessary for allergic reactions to release inflammatory mediators that could contribute to CFS symptoms.Objective
To investigate the effect of isoflavones on the action of polyinosinic:polycytidylic acid (poly(I:C)), with or without swim stress, on mouse locomotor activity and inflammatory mediator expression, as well as on human MC activation.Methods
Female C57BL/6 mice were randomly divided into four groups: (a) control/no-swim, (b) control/swim, (c) poly(I:C)/no swim, and (d) poly(I:C)/swim. Mice were provided with chow low or high in isoflavones for 2 weeks prior to ip injection with 20 mg/kg poly(I:C) followed or not by swim stress for 15 minutes. Locomotor activity was monitored overnight and animals were sacrificed the following day. Brain and skin gene expression, as well as serum levels, of inflammatory mediators were measured. Data were analyzed using the non-parametric Mann-Whitney U-test.ResultsPoly(I:C)-treated mice had decreased locomotor activity over 24 hours, and increased serum levels of TNF-¿, IL-6, KC (IL-8/CXCL8 murine homolog), CCL2,3,4,5, CXCL10, as well as brain and skin gene expression of TNF, IL-6, KC (Cxcl1, IL8 murine homolog), CCL2, CCL4, CCL5 and CXCL10. Histidine decarboxylase (HDC) and NT expression were also increased, but only in the skin, over the same period. High isoflavone diet reversed these effects.Conclusion
Poly(I:C) treatment decreased mouse locomotor activity and increased serum levels and brain and skin gene expression of inflammatory mediators. These effects were inhibited by isoflavones that may prove useful in CFS.
... As an ancient traditional drug, it has been used for ages as a rejuvenator [8,9] and for treating a number of pathological conditions including tumors and pimples, bone fractures, diseases of skin, neuralgia, arthritis, inflammation, asthma, angina, gastrointestinal, and genitourinary diseases [9]. Shilajit comprises of 60-80 % humus along with other organic components such as benzoic acid, hippuric acid, fatty acid, ichthyol, ellagic acid, resin, triterpenes, sterol, aromatic carboxylic acid, 3,4-benzocoumarins, amino acids, and phenolic lipids [10,11]. The approximate composition of Asian mumie is 20 % minerals, 15 % protein, 5 % lipids, 5 % steroids, and also some carbohydrates, alkaloids, and amino acids [12]. ...
This study assessed the effects of mumie (shilajit) pre-treatment, a traditional drug which is well known in the ancient medicine of both east and west, on cardiac performance of rats subjected to myocardial injury. Animals were divided into control, M250, and M500 (received mumie at dosages of 250 and 500 mg/kg/day, orally for 7 days, respectively) main groups each consisting of two subgroups-with and without heart injury. On the 6th and 7th days, isoproterenol (ISO) (85 mg/kg i.p.) was injected (s.c.) to half of the animal subgroups to induce myocardial damage. On the 8th day, after hemodynamic parameter recordings, hearts were removed for further evaluation. Mumie pre-treatment had no significant effects on hemodynamic and cardiac indices of normal animals. When the cardiac injury was induced, mumie maintained the ±dp/dt maximum, attenuated the serum cardiac troponin I, and reduced the severity of cardiac lesions. Despite the mild positive effects of mumie on total antioxidant capacity and lipid proxidation index, no significant difference was observed among animal groups. The findings suggest the prominent cardioprotective effect of mumie against destructive effects of ISO. It seems that other mechanisms than reinforcements of antioxidant system are involved in this beneficial effect.
Chronic fatigue syndrome (CFS) has a high incidence due to the increased pressure of daily life and work in modern society. Our previous clinical studies have found the effects of electroacupuncture (EA) on CFS patients, however, the mechanism of EA on CFS is still unknown. In this study, we investigated the effects of EA on cardiac function in a CFS mouse model to explore its underlying mechanism. The mice were randomly divided into three groups: control, CFS, and CFS mice receiving EA (CFS + EA). After behavioral assessments and echocardiographic measurement, blood and heart tissue of the mice were collected for biochemical tests, and then we evaluated the effects of EA on the CFS mouse model when nitric oxide (NO) levels were enhanced by l‐arginine. The results showed that EA ameliorated the injured motor and cardiac function. Meanwhile, EA also inhibited increased expression of inducible nitric oxide synthase (iNOS) at heart tissue and the serum NO levels in mice subjected to sustained forced swimming stress. Furthermore, the NO level in serum increased with l‐arginine administration, which blocked the effects of EA on CFS mice. This study suggested that EA could improve the motor function and cardiac function in CFS mice and its effects may be associated with the down‐regulation of iNOS/NO signaling. 由于现代社会日常生活和工作的压力逐渐增加, 慢性疲劳综合征(CFS)出现了较高的发病率。我们之前的临床研究已经证明了电针(EA)对CFS患者有明显的治疗效果。然而, EA治疗CFS的机制仍不清楚。在这项研究中, 我们考察了EA对CFS小鼠模型心脏功能的作用以探索其潜在机制。将小鼠随机分为三组:空白对照组, CFS组以及CFS小鼠接受EA干预(CFS + EA)组。在行为学评估和心脏超声测量后, 收集小鼠的血液和心脏组织进行生化检测, 随后我们还评估了当L‐精氨酸上调一氧化氮(NO)水平的情况下, EA干预CFS小鼠模型的作用。结果表明, EA改善了运动和心脏功能。同时, EA可抑制持续强迫游泳应激小鼠的心脏诱导型一氧化氮合酶(iNOS)表达和血清NO水平。此外, L‐精氨酸给药可增加血清中的NO水平, 从而阻断了EA对CFS小鼠的作用。本研究表明, 电针可改善CFS小鼠的运动功能和心脏功能, 这可能与对iNOS/NO信号的下调有关。.
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a serious, complex, and highly debilitating long-term illness. People with ME/CFS are typically unable to carry out their routine activities. Key hallmarks of the disease are neurological and gastrointestinal impairments accompanied by pervasive malaise that is exacerbated after physical and/or mental activity. Currently, there is no validated cure of biomarker signature for this illness. Impaired tryptophan (TRYP) metabolism is thought to play significant role in the pathobiology of ME/CFS. TRYP is an important precursor for serotonin and the essential pyridine nucleotide nicotinamide adenine dinucleotide (NAD+). TRYP has been associated with the development of some parts of the brain responsible for behavioural functions. The main catabolic route for TRYP is the kynurenine pathway (KP). The KP produces NAD+ and several neuroactive metabolites with neuroprotective (i.e., kynurenic acid (KYNA)) and neurotoxic (i.e., quinolinic acid (QUIN)) activities. Hyperactivation of the KP, whether compensatory or a driving mechanism of degeneration can limit the availability of NAD+ and exacerbate the symptoms of ME/CFS. This review discusses the potential association of altered KP metabolism in ME/CFS. The review also evaluates the role of the patient's gut microbiota on TRYP availability and KP activation. We propose that strategies aimed at raising the levels of NAD+ (e.g., using nicotinamide mononucleotide and nicotinamide riboside) may be a promising intervention to overcome symptoms of fatigue and to improve the quality of life in patients with ME/CFS. Future clinical trials should further assess the potential benefits of NAD+ supplements for reducing some of the clinical features of ME/CFS.
Bu çalışmanın amacı, bal arısı (Apis melifera) sperma sulandırıcısına katılan farklı dozlardaki (%0, 5, 10, 15, 20) shilajitin (S) motilite, plazma membran bütünlüğü ve akrozom bütünlüğü parametreleri üzerine etkilerini belirlemektir. Çalışmada, 5 grup ve her bir grupta 5’er adet koloni olmak üzere toplam 25 adet koloni kullanıldı. Kontrol grubu (S-0), bal arısı sulandırıcısına shilajit ilavesi yapılmayan grubu oluşturmaktadır. S-1, S-2, S-3 ve S-4 grupları ise bal arısı sulandırıcısına sırasıyla %5, 10, 15 ve 20 shilajit ilaveli gruplar olarak oluşturuldu. Gün aşırı beslenen yaklaşık 25 adet koloni içerisinde 14-21 günlük yaşta bulunan yaklaşık 400 erkek arıdan sperma toplandı ve sıvı azot buharında dondurularak, -196°C sıvı azot içerisinde yaklaşık 7 ay süre ile saklandı. Bu süre sonunda dondurulan numuneler 37°C’ de çözdürülerek ilgili parametreler yönünden incelendi. Diğer gruplar ile karşılaştırıldığında, S-3 ve S-4 gruplarının sperma motilite değerini arttırdığı (p<0.05); S-2, S-3 ve S-4 gruplarının plazma membran bütünlüğünü arttırdığı tespit edildi. Sulandırıcıya farklı dozlarda shilajit ilave edilen gruplarda, kontrol grubuna kıyasla akrozom bütünlüğününün önemli derecede korunduğu belirlendi (p<0.01). Sonuç olarak, bal arısı sulandırıcısına farklı dozlardaki shilajitin spermatolojik parametreler üzerine olumlu etkileri olduğu saptandı.
Shilajit is used commonly as Ayurvedic medicine worldwide which is Rasayana herbo-mineral substance and consumed to restore the energetic balance and to prevent diseases like cognitive disorders and Alzheimer. Locally, Shilajit is applied for patients diagnosed with bone fractures. For safety of the patients, the elemental analysis of Shilajit is imperative to evaluate its nutritional quality as well as contamination from heavy metals. The elemental composition of Shilajit was conducted using three advanced analytical techniques (LIBS, ICP, and EDX). For the comparative studies, the two Shilajit kinds mostly sold globally produced in India and Pakistan were collected. Our main focus is to highlight nutritional eminence and contamination of heavy metals to hinge on Shilajit therapeutic potential. In this work, laser-induced breakdown spectroscopy (LIBS) was applied for qualitative and quantitative analysis of the Shilajit. Our LIBS analysis revealed that Shilajit samples composed of several elements like Ca, S, K, Mg, Al, Na, Sr, Fe, P, Si, Mn, Ba, Zn, Ni, B, Cr, Co, Pb, Cu, As, Hg, Se, and Ti. Indian and Pakistani Shilajits were highly enriched with Ca, S, and K nutrients and contained Al, Sr, Mn, Ba, Zn, Ni, B, Cr, Pb, As, and Hg toxins in amounts that exceeded the standard permissible limit. Even though the content of most elements was comparable among both Shilajits, nutrients, and toxins, in general, were accentuated more in Indian Shilajit with the sole detection of Hg and Ti. The elemental quantification was done using self-developed calibration-free laser-induced breakdown spectroscopy (CF-LIBS) method, and LIBS results are in well agreement with the concentrations determined by standard ICP-OES/MS method. To verify our results by LIBS and ICP-OES/MS techniques, EDX spectroscopy was also conducted which confirmed the presence above mentioned elements. This work is highly significant for creating awareness among people suffering due to overdose of this product and save many human lives.
Background:
Mummy (Iranian pure shilajit) is a remedy with possessing anti-inflammatory, antioxidant and anticancer activities. This study aimed to examine mummy effects on epithelial-mesenchymal transition (EMT) and invasiveness of MCF-7 and MDA-MB-231 breast cancer (BC) cell lines with underlying its mechanism.
Materials and methods:
The dose-dependent inhibitory effect of the mummy on cell proliferation in vitro was determined using the MTT assay. Flow cytometry and 4',6-diamidino-2-phenylindole dihydrochloride staining were respectively used for quantitative and qualitative analysis of cellular apoptosis, and gene expression analysis was conducted using real-time PCR.
Results:
MDA-MB-231 showed more sensitivity than the MCF-7 cell line to the anticancer activity of mummy, while mummy did not exhibit significant cell cytotoxicity against human normal cells (MCF-10A). The gene expression profile demonstrated a significant decrease in TGF-β1, TGF-βR1, TWIST1, NOTCH1, CTNNB1, SRC along with an increase in E-cadherin mRNA levels in mummy treated cells compared to the untreated control group (P≤0.05).
Conclusion:
Mummy triggers inhibition of EMT and metastasis in breast cancer cells mainly through the downregulation of TGFβ1 activity, and more studies required to find its specific anticancer activity with details.
Chronic fatigue syndrome (CFS) is a disorder characterized by persistent and relapsing fatigue along with long-lasting and debilitating fatigue, myalgia, cognitive impairment, and many other common symptoms. The present study was conducted to explore the protective effect of hemin on CFS in experimental mice. Male albino mice were subjected to stress-induced CFS in a forced swimming test apparatus for 21 days. After animals had been subjected to the forced swimming test, hemin (5 and 10 mg/kg; i.p.) and hemin (10 mg/kg) + tin(IV) protoporphyrin (SnPP), a hemeoxygenase-1 (HO-1) enzyme inhibitor, were administered daily for 21 days. Various behavioral tests (immobility period, locomotor activity, grip strength, and anxiety) and estimations of biochemical parameters (lipid peroxidation, nitrite, and GSH), mitochondrial complex dysfunctions (complexes I and II), and neurotransmitters (dopamine, serotonin, and norepinephrine and their metabolites) were subsequently assessed. Animals exposed to 10 min of forced swimming session for 21 days showed a fatigue-like behavior (as increase in immobility period, decreased grip strength, and anxiety) and biochemical alteration observed by increased oxidative stress, mitochondrial dysfunction, and neurotransmitter level alteration. Treatment with hemin (5 and 10 mg/kg) for 21 days significantly improved the decreased immobility period, increased locomotor activity, and improved anxiety-like behavior, oxidative defense, mitochondrial complex dysfunction, and neurotransmitter level in the brain. Further, these observations were reversed by SnPP, suggesting that the antifatigue effect of hemin is HO-1 dependent. The present study highlights the protective role of hemin against experimental CFS-induced behavioral, biochemical, and neurotransmitter alterations.
Background
Shilajit (mumie), a natural multi-component herbomineral ethnomedicinal food, is used as a traditional medicine for enhancing the quality of life and for management of health ailments in many countries of the world. Use of Shilajit as an adaptogen, aphrodisiac, rejuvenator and anti-aging substance is mentioned in many ancient texts. This review aims to provide comprehensive insights into its biochemical aspects, microbial role in biosynthesis, bioactivities and to establish correlation between traditional uses and scientifically validated research findings.
Methods
Scientific literature and ethnopharmacological information were compiled from the published peer-reviewed articles, unpublished materials, thesis, books, patent databases, clinical trial registries and from the websites of research councils of traditional medicine. The scientific databases, thesis repositories and books databases were searched with keywords Shilajit, mumie, mumijo, salajeet, asphaltum, fulvic acid, dibenzo-alpha-pyrones etc.
Results
Scientifically validated research and ancient texts suggest multifaceted benefits of Shilajit. It is endowed with anti-stress, memory and energy enhancing, antioxidant, anti-inflammatory, antidiabetic, spermatogenic, neuroprotective, antiulcer and wound healing activities. These pharmacological effects are mainly attributed to the presence of humic acid, fulvic acid, dibenzo-α-pyrones, dibenzo- α-pyrones chromoproteins and trace elements.
Conclusion
This review summarizes the traditional importance of Shilajit for the treatment and prevention of several acute and chronic diseases and health ailments. Despite numerous health claims, there are still major gaps in our understanding of its mechanism of action, variability in efficacy and toxicity profile. Therefore, a coordinated interdisciplinary approach is needed to establish the underlying mechanisms of action, comprehensive toxicological profile, pharmacokinetics parameters and effects on different organ systems. Regulatory and governmental impetus to basic and clinical research, safety testing and formulations quality control is warranted.
Aim of the study:
To assess the effectiveness of TRPV1 modulators in animal model of Chronic fatigue syndrome (CFS). To assess central and peripheral behavioral activity of TRPV1 modulators.
Material and methods:
CFS was induced by forcing the rats to swim for 10min for 21 consecutive days. The rats were treated with capsaicin (TRPV1 agonist, 2.5mg/kg) and n-tert-butylcyclohexanol (TRPV1 antagonist, 10mg/kg) for 21days 30min before the exposure to stress procedure. The behavioral consequence of CFS was measured in terms of immobility time, grip strength, locomotor activity, and anxiety level using Rota rod, Actophotometer, and Elevated plus maze model respectively. The other parameters include Plasma corticosterone, adrenal gland and spleen weight, complete blood count, blood urea niterogen (BUN), Lactate dehydrogenase (LDH), Lipid peroxidation, catalase and reduced glutathione (GSH).
Results and discussion:
TRPV1 modulators reversed (p<0.05) the increase in immobility period, anxiety, spleen weight, BUN and LDH levels, and MDA levels along with decrease in grip strength, locomotor activity, plasma corticosterone, adrenal gland weight, catalase, and GSH. There was also significant increase in total WBC count when compared with the disease control group. The reversal was attributed to modulation of HPA axis, oxidative stress, anaerobic respiration product, muscle degradation product.
Conclusion:
The present study reveals the effectiveness of n-tert-butylcyclohexanol and capsaicin against chronic fatigue syndrome. The mechanism of action can be attributed to inhibition of TRPV1 channel and thereby modulating pain perception, neuroendocrine function, oxidative stress and immune function.
Shilajit is a widely used natural herbo-mineral in Ayurveda, the traditional Indian system of medicine. It is a sticky brown to blackish physiologically active organic matter exuded from steep rocks in many mountain ranges of the world, especially the Himalayas of the Indian subcontinent. Shilajit is composed of humus and organic plant material, and its active constituents include dibenzo-alpha-pyrones, dibenzo-alpha-pyrone-chromoproteins, and fulvic acids. It exhibits antioxidant, immunomodulatory, anti-inflammatory, adaptogenic, and antidyslipidemic activities. It has cholinergic and parasympathomimetic effects. Studies conducted in animal models tend to support its use as a “revitalizer,” enhancing physical performance and relief from fatigue with enhanced ATP production. Further systematic research is needed to elucidate the exact mechanism of action of shilajit as a rejuvenator. Studies involving animals and humans indicate that the use of shilajit may be safe and free from adverse effects. Additional well-controlled studies on standardized products are needed in humans and animals.
Shilajit (mumie; moomiyo, mummiyo) has been used for a wide variety of illnesses and conditions for many years. However, relatively few well-controlled human studies have been conducted on the effects of shiliajit, although a growing number of studies have been published in recent years involving animal and in vitro systems. The safety of shilajit is well documented based on animal and human studies. Various research studies indicate that shilajit exhibits antioxidant, anti-inflammatory, adaptogenic, immunomodulatory, and anti-dyslipidemic properties. Animal and human studies indicate that shilajit enhances spermatogenesis. Furthermore, animal and human data support its use as a 'revitalizer', enhancing physical performance and relieving fatigue with enhanced production of ATP. Key constituents in shilajit responsible for these effects appear to be dibenzo-α-pyrones and fulvic acid and their derivatives. Various mechanistic studies provide support for the above observed effects. Additional well-controlled human and animal studies involving the use of standardized products are needed. Copyright © 2013 John Wiley & Sons, Ltd.
Chronic fatigue syndrome (CFS) is an illness characterized by persistent and relapsing fatigue, often accompanied by numerous symptoms involving various body systems. The etiology of CFS remains unclear; however, a number of studies have shown that oxidative stress may be involved in its pathogenesis. In the present study, a mouse model of CFS was used in which mice were forced to swim for one 6-minute session on each day for 15 days and the immobility period was recorded. There was a significant increase in immobility period in saline-treated mice on successive days. Intraperitoneal treatment with the potent antioxidants carvedilol (5 mg/kg) and melatonin (5 mg/kg) produced a significant reduction in immobility period. Similar results were observed with herbal preparations administered orally: Withania somnifera (100 mg/kg), quercetin (50 mg/kg), and St. John's wort (Hypericum perforatum L., 10 mg/kg). Biochemical analysis revealed that chronic swimming significantly induced lipid peroxidation and decreased glutathione (GSH) levels in the brains of mice. The rats also showed decreased levels of antioxidant defense enzymes, superoxide dismutase (SOD), and catalase. Co-administration of antioxidants carvedilol, melatonin, W. somnifera, quercetin or St. John's wort significantly reduced lipid peroxidation and restored the GSH levels decreased by chronic swimming in mice. Further, the treatment increased levels of SOD in the forebrain and of catalase. The findings strongly suggest that oxidative stress plays a significant role in the pathophysiology of CFS and that antioxidants could be useful in the treatment of CFS.
The effect of Shilajit was investigated for putative nootropic and anxiolytic activity, and its
effect on rat brain monoamines using Charles Foster strain albino rats. Nootropic activity
was assessed by passive avoidance learning and active avoidance learning acquisition and
retention. Anxiolytic activity was evaluated by the elevated plus-maze technique. Rat brain
monoamines and monoamine metaboliteswere estimated bya HPLC technique. The results
indicated that Shilajit had significant nootropic and anxiolytic activity. The biochemical
studies indicated that acute treatment with Shilajit had insignificant effects on rat brain
monoamine and monoamine metabolite levels. However, following subacute (5days) treatment,
there was decrease in 5-hydroxytryptamine and 5-hydroxyindole acetic acid concentrations
and an increase in the levels of dopamine, homovanillic acid and
3.4-dihydroxyphenyl-acetic acid concentrations, with insignificant effects on noradrenaline
and 3-methoxy-4- hydroxyphenylethylene glycol levels. The observed neurochemical effects
induced by Shilajit, indicating a decrease in rat brain 5-hydroxytryptamine turnover, associated
with an increase in dopaminergic activity, helps to explain the observed nootropic and
anxiolytic effects of the drug.
A simplified procedure has been developed for the extraction and estimation of corticosterone from rat serum, tissues and mitochondria. The suitably diluted samples were treated with freshly prepared chloroform: methanol mixture (2:1, v/v) and then extracted directly with the chloroform. Almost quantitative recoveries (90% and above) were obtained with the present method, compared to poor recoveries (65-81%) and variable results obtained by earlier procedure. Quantification of corticosterone content in tissues, such as liver, brain and heart, and in the mitochondria indicated significant concentration of corticosterone in tissues and mitochondria, as compared to the serum. The presence of corticosterone in the mitochondria suggests that the hormone may play a role in regulation of mitochondrial gene expression and/or their turnover.
• The complexities of the chronic fatigue syndrome and the methodologic problems associated with its study indicate the need for a comprehensive, system atic, and integrated approach to the evaluation, classi fication, and study of persons with this condition and other fatiguing illnesses. We propose a conceptual framework and a set of guidelines that provide such an approach. Our guidelines include recommendations for the clinical evaluation of fatigued persons, a revised case definition of the chronic fatigue syndrome, and a strategy for subgrouping fatigued persons in formal investigations.
Major and biologically most explored components of natural organic matter (NOM) are humic acid (HA) and fulvic acid (FA). We have explored rock shilajit as a source of NOM. On the other hand carbamazepine (CBZ) is a well known anticonvulsant drug and has a limited accessibility to brain. Bioavailability and pharmacokinetic profiles of CBZ have been improved by complexation and different techniques also.Present study has assessed the comparative abilities of FA and HA as complexing agent for CBZ in order to enhance pharmacokinetic profile of CBZ and accessibility to the brain. These two complexing agents have been compared on various indices such as their abilities to cause complexation and enhance solubility, permeability and dissolution. The present study also compared pharmacodynamic and biochemical profiles after oral administration of complexes. With the help of various pharmaceutical techniques such as freeze drying, physical mixture, kneading and solvent evaporation, two molar ratios (1:1 and 1:2) were selected for complexation and evaluated for conformational analysis (molecular modeling). Complex formed was further characterized by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), mass spectroscopy and X-ray diffraction (XRD).Preclinical study on rodents with CBZ–HA and CBZ–FA has yielded appreciable results in terms of their anticonvulsant and antioxidants activities. However, CBZ–HA (1:2) demonstrated better result than any other complex.
We have investigated the effect of shilajit on lipid peroxidation and glutathione content in rat liver homogenate. It inhibited lipid peroxidation induced by cumene hydroperoxide and ADP/Fe++ complex in a dose dependent manner. It also reduced the rate of oxidation of reduced glutathione content and inhibited ongoing lipid peroxidation, induced by these agents immediately after its addition to the incubation system.
This paper describes the problems of measuring the allosteric ATP-inhibition of cytochrome c oxidase (CcO) in isolated mitochondria. Only by using the ATP-regenerating system phosphoenolpyruvate and pyruvate kinase full ATP-inhibition of CcO could be demonstrated by kinetic measurements. The mechanism was proposed to keep the mitochondrial membrane potential (DeltaPsi(m)) in living cells and tissues at low values (100-140 mV), when the matrix ATP/ADP ratios are high. In contrast, high DeltaPsi(m) values (180-220 mV) are generally measured in isolated mitochondria. By using a tetraphenyl phosphonium electrode we observed in isolated rat liver mitochondria with glutamate plus malate as substrates a reversible decrease of DeltaPsi(m) from 233 to 123 mV after addition of phosphoenolpyruvate and pyruvate kinase. The decrease of DeltaPsi(m) is explained by reversal of the gluconeogenetic enzymes pyruvate carboxylase and phosphoenolpyruvate carboxykinase yielding ATP and GTP, thus increasing the matrix ATP/ADP ratio. With rat heart mitochondria, which lack these enzymes, no decrease of DeltaPsi(m) was found. From the data we conclude that high matrix ATP/ADP ratios keep DeltaPsi(m) at low values by the allosteric ATP-inhibition of CcO, thus preventing the generation of reactive oxygen species which could generate degenerative diseases. It is proposed that respiration in living eukaryotic organisms is normally controlled by the DeltaPsi(m)-independent "allosteric ATP-inhibition of CcO." Only when the allosteric ATP-inhibition is switched off under stress, respiration is regulated by "respiratory control," based on DeltaPsi(m) according to the Mitchell Theory.
The safety and spermatogenic activity of processed Shilajit (PS) were evaluated in oligospermic patients. Initially, 60 infertile male patients were assessed and those having total sperm counts below 20 million ml(-1) semen were considered oligospermic and enrolled in the study (n = 35). PS capsule (100 mg) was administered twice daily after major meals for 90 days. Total semenogram and serum testosterone, luteinising hormone and follicle-stimulating hormone were estimated before and at the end of the treatment. Malondialdehyde (MDA), a marker for oxidative stress, content of semen and biochemical parameters for safety were also evaluated. Twenty-eight patients who completed the treatment showed significant (P < 0.001) improvement in spermia (+37.6%), total sperm count (+61.4%), motility (12.4-17.4% after different time intervals), normal sperm count (+18.9%) with concomitant decrease in pus and epithelial cell count compared with baseline value. Significant decrease of semen MDA content (-18.7%) was observed. Moreover, serum testosterone (+23.5%; P < 0.001) and FSH (+9.4%; P < 0.05) levels significantly increased. HPLC chromatogram revealed inclusion of PS constituents in semen. Unaltered hepatic and renal profiles of patients indicated that PS was safe at the given dose. The present findings provide further evidence of the spermatogenic nature of Shilajit, as attributed in Ayurvedic medicine, particularly when administered as PS.
This study aims to improve the health of patients suffering from chronic fatigue syndrome (CFS) by interventions based on the biochemistry of the illness, specifically the function of mitochondria in producing ATP (adenosine triphosphate), the energy currency for all body functions, and recycling ADP (adenosine diphosphate) to replenish the ATP supply as needed. Patients attending a private medical practice specializing in CFS were diagnosed using the Centers for Disease Control criteria. In consultation with each patient, an integer on the Bell Ability Scale was assigned, and a blood sample was taken for the "ATP profile" test, designed for CFS and other fatigue conditions. Each test produced 5 numerical factors which describe the availability of ATP in neutrophils, the fraction complexed with magnesium, the efficiency of oxidative phosphorylation, and the transfer efficiencies of ADP into the mitochondria and ATP into the cytosol where the energy is used. With the consent of each of 71 patients and 53 normal, healthy controls the 5 factors have been collated and compared with the Bell Ability Scale. The individual numerical factors show that patients have different combinations of biochemical lesions. When the factors are combined, a remarkable correlation is observed between the degree of mitochondrial dysfunction and the severity of illness (P<0.001). Only 1 of the 71 patients overlaps the normal region. The "ATP profile" test is a powerful diagnostic tool and can differentiate patients who have fatigue and other symptoms as a result of energy wastage by stress and psychological factors from those who have insufficient energy due to cellular respiration dysfunction. The individual factors indicate which remedial actions, in the form of dietary supplements, drugs and detoxification, are most likely to be of benefit, and what further tests should be carried out.
An experimental model of chronic fatigue syndrome (CFS) is utilized for evaluation of antidepressant, anti-stress effects, wherein the rat is forced to swim in water for 15 min/day on 21 consecutive days. Rats were divided into stressed control, stressed plus standard drug (Panax ginseng) and stressed plus 200 and 500 mg/kg of test drug, i.e., Nardostachys jatamansi extract (NJE) given orally. The immobility during each 5 min periods of 0-5, 5-10 and 10-15 min of stress were noted. Similarly the climbing (struggling) behaviour was noted in the above four groups of rats in intervals of 5 min. The locomotor activity and also the anxiety state in animals were evaluated in an elevated plus maze after CFS in all the four groups. There was a significant increase in despair behaviour and anxiety in stressed control animals on successive days of CFS. Locomotor activity gradually decreased in stressed control group. Treatment with NJE (200 and 500 mg/kg) significantly reversed both paradigms. Biochemical analysis showed that CFS significantly increased lipid peroxidation, nitrite and superoxide dismutase levels and decreased catalase level in rat brain. Administration of NJE (200 and 500 mg/kg) tended to normalize both augmented lipid peroxidation, nitrite, superoxide dismutase activities and catalase level significantly. NJE per se has an antioxidant effect. The results indicate that CFS may lead to oxidative stress, which is mitigated by NJE and so its antioxidant property may be responsible for anti-stress effect of NJE.
A new automated system for the analysis of nitrate via reduction with a high-pressure cadmium column is described. Samples of urine, saliva, deproteinized plasma, gastric juice, and milk can be analyzed for nitrate, nitrite, or both with a lower limit of detection of 1.0 nmol NO3− or NO2−/ml. The system allows quantitative reduction of nitrate and automatically eliminates interference from other compounds normally present in urine and other biological fluids. Analysis rate is 30 samples per hour, with preparation for most samples limited to simple dilution with distilled water. The application of gas chromatography/mass spectrometry for the analysis of 15NO3− in urine after derivatization to 15NO2-benzene is also described.
This study aimed to determine symptom patterns in patients with chronic fatigue syndrome (CFS), in summer and winter. Comparison data for patients with seasonal affective disorder (SAD) were used to evaluate seasonal variation in mood and behavior, atypical neurovegetative symptoms characteristic of SAD, and somatic symptoms characteristic of CFS. Rating scale questionnaires were mailed to patients previously diagnosed with CFS. Instruments included the Personal Inventory for Depression and SAD (PIDS) and the Systematic Assessment for Treatment Emergent Effects (SAFTEE), which catalogs the current severity of a wide range of somatic, behavioral, and affective symptoms. Data sets from 110 CFS patients matched across seasons were entered into the analysis. Symptoms that conform with the Centers for Disease Control and Prevention (CDC) case definition of CFS were rated as moderate to very severe during the winter months by varying proportions of patients (from 43% for lymph node pain or enlargement, to 79% for muscle, joint, or bone pain). Fatigue was reported by 92%. Prominent affective symptoms included irritability (55%), depressed mood (52%), and anxiety (51%). Retrospective monthly ratings of mood, social activity, energy, sleep duration, amount eaten, and weight change showed a coherent pattern of winter worsening. Of patients with consistent summer and winter ratings (n = 73), 37% showed high global seasonality scores (GSS) > or = 10. About half this group reported symptoms indicative of major depressive disorder, which was strongly associated with high seasonality. Hierarchical cluster analysis of wintertime symptoms revealed 2 distinct clinical profiles among CFS patients: (a) those with high seasonality, for whom depressed mood clustered with atypical neurovegetative symptoms of hypersomnia and hyperphagia, as is seen in SAD; and (b) those with low seasonality, who showed a primary clustering of classic CFS symptoms (fatigue, aches, cognitive disturbance), with depressed mood most closely associated with irritability, insomnia, and anxiety. It appears that a subgroup of patients with CFS shows seasonal variation in symptoms resembling those of SAD, with winter exacerbation. Light therapy may provide patients with CFS an effective treatment alternative or adjunct to antidepressant drugs.
Preparations of rat-liver mitochondria catalyze the oxidation of exogenous NADH by added cytochrome c or ferricyanide by a reaction that is insensitive to the respiratory chain inhibitors, antimycin A, amytal, and rotenone, and is not coupled to phosphorylation. Experiments with tritiated NADH are described which demonstrate that this "external" pathway of NADH oxidation resembles stereochemically the NADH-cytochrome c reductase system of liver microsomes, and differs from the respiratory chain-linked NADH dehydrogenase. Enzyme distributation data are presented which substantiate the conclusion that microsomal contamination cannot account for the rotenone-insensitive NADH-cytochrome c reductase activity observed with the mitochondria. A procedure is developed, based on swelling and shrinking of the mitochondria followed by sonication and density gradient centrifugation, which permits the separation of two particulate subfractions, one containing the bulk of the respiratory chain components, and the other the bulk of the rotenone-insensitive NADH-cytochrome c reductase system. Morphological evidence supports the conclusion that the former subfraction consists of mitochondria devoid of outer membrane, and that the latter represents derivatives of the outer membrane. The data indicate that the electron-transport system associated with the mitochondrial outer membrane involves catalytic components similar to, or identical with, the microsomal NADH-cytochrome b(5) reductase and cytochrome b(5).
Kainic acid (KA), a potent central excitotoxin, may elicit neuronal death via generation of reactive oxygen species (ROS). The present study was undertaken to further characterize KA neurotoxicity and its relationship to ROS production and mitochondrial dysfunction. Exposure of rat cerebellar granule neurons at 14 days in vitro to 0.5 mM KA for 30 min resulted in the death of 53% of cells 24 h later. ROS production, evaluated by 2',7'-dichlorofluorescein diacetate, increased in KA-treated granule neurons. Resolution of mitochondrial oxidative phosphorylation enzymes by blue native polyacrylamide gel electrophoresis, followed by histochemical staining, showed that KA induced a strong decrease (-40%, P
The mitochondrion plays a pivotal role in energy metabolism in eukaryotic cells. The electrochemical potential across the mitochondrial inner membrane is regulated to cope with cellular energy needs and thus reflects the bioenergetic state of the cell. Traditional assays for mitochondrial membrane potential are not amenable to high-throughput drug screening. In this paper, I describe a high-throughput assay that measures the mitochondrial membrane potential of living cells in 96- or 384-well plates. Cells were first treated with test compounds and then with a fluorescent potentiometric probe, the cationic-lipophilic dye tetramethylrhodamine methyl ester (TMRM). The cells were then washed to remove free compounds and probe. The amount of TMRM retained in the mitochondria, which is proportional to the mitochondrial membrane potential, was measured on an LJL Analyst fluorescence reader. Under optimal conditions, the assay measured only the mitochondrial membrane potential. The chemical uncouplers carbonylcyanide m-chlorophenyl hydrazone and dinitrophenol decreased fluorescence intensity, with IC(50) values (concentration at 50% inhibition) similar to those reported in the literature. A Z' factor of greater than 0.5 suggests that this cell-based assay can be adapted for high-throughput screening of chemical libraries. This assay may be used in screens for drugs to treat metabolic disorders such as obesity and diabetes, as well as cancer and neurodegenerative diseases.
Withania somnifera and Panax ginseng are well-known for their energy augmenting effect. In this study, Withania somnifera and Panax ginseng standardized extracts were evaluated for their energy augmenting activity in an experimental model using the forced swimming test (FST) in Swiss albino mice. Withania somnifera standardized extract (WSE) dose dependently attenuated ATP-depletion and other energy related indices during both short term (7 days) and long term (30 days) treatment periods. The treatment doses were 50 mg/kg and 100 mg/kg. Panax ginseng (PGE), on the other hand, did not elicit similar energy-restoring effect when compared with that of WSE at the lower dose level and on prolonged administration (50 mg/kg for 30days). This latter finding would seem to project WSE as a better adaptogen. Also, WSE may be considered as a better agent for stress management in view of the well documented adverse effects of Ginseng (known as the Ginseng abuse syndrome). To achieve this objective, the need for standardization of Withania somnifera extract is emphasized.
The decrease in efficiency of mitochondria in generating energy currency (ATP) in animals and in humans is associated with aging (geriatric problems) and oxidative stress. This deficiency has a link with the systemic deficiencies of coenzyme Q10 (CoQ10) concentration and of two of its endogenous functional associates, namely, 3-hydroxydibenzo-α-pyrone (3-OH-DBP) and 3,8- dihydroxydibenzo-α-pyrone [3,8-(OH)2-DBP]. Mitochondrial targeting of the two DBPs, isolated from shilajit (the supervitalizer of Ayurveda), and of CoQ10 could be formidable strategies to augment antioxidant defense and energy generating elements for restoring normal functions of mitochondria. DBPs, as also their fatty-acyl and amino-acyl conjugates, occur in animal mitochondria and in blood where they act in tandem with CoQ10 in the electron transport chain. Administration of CoQ10 alone, in mitochondrial deficiency states, therefore, could not restore normal mitochondrial functions. The concomitant targeting of DBPs and CoQ10 to mitochondria would augment energy (ATP) synthesis and protect redox states of CoQ10 from oxidative degradation. The present findings adduce evidence of augmentation of the concentrations of DBPs and CoQ10 in mitochondria when administered, from exogenous sources, through intra-peritoneal/oral route. Their probable mechanism of action would involve the three redox states of DBPs (reduced form, semiquinone radical and quinone form) and similar redox states of CoQ10 as a measure to restore normal energy synthesizing ability of mitochondria.
The present chapter discusses metal-containing flavoprotein dehydrogenases. Flavoproteins are involved in a large variety of key metabolic reactions in all forms of life. They catalyze over a potential span of several hundred millivolts oxidation–reduction reactions involving alkanes, alkenes, alcohols, aldehydes, ketones, inorganic and organic acids, amines, thiols, disulfides, quinones, nicotinamide-adenine dinucleotides, purines, pyrimidines, pteridines, and transition metal complexes. They can also catalyze one- and two-electron reduction of molecular oxygen. Many flavoproteins contain metal such as iron, molybdenum, and zinc. The combination of flavin and metal often serves to adjust electron transfer between single-electron and double-electron donors and acceptors. Multiple-electron reduction of an acceptor without detectable loss of intermediates is achieved by the device of having multiple flavins and metals in the same enzyme molecule. The chapter discusses the enzymes that are respiratory chain-linked NADH dehydrogenases, succinate dehydrogenases, L-glycerol-3-phosphate dehydrogenase, choline dehydrogenase, and L (+)-lactate.
Shilajit is an asphalt-like substance found embedded in rocky sediments in the Himalayas in western Nepal at altitudes between 2500-5000 m. It is popularly used in Nepal as a tonic. Chemical analysis of shilajit revealed that two-thirds by weight of this medicinal material was extractable by warm 50% alcohol. Repeated crystallization of the hydroalcoholic extract has led to the isolation of crystals, which were subsequently identified as calcium benzoate. The antiseptic properties of benzoates may account for the antiseptic effects of shilajit in places where hygiene remains at a low level.
The chemical polemics in the reported literature on shilajit are resolved. This study shows that humification of latex and resin-bearing plants is responsible for the major organic mass (80-85%) of shilajit. The low mol. wt. chemical markers (&lo%), viz. aucuparins, oxygenated dibenzo-K -pyrones and triterpenic acids of the tirucallane type (free and conjugated), occurring in the core structure of shilajit humus, are the major active constituents of Himalayan shilajit. The therapeutic effects of shilajit are the consequences of hormonal control and regulation of immunity.
Patients with functional somatic syndromes such as fibromyalgia (FM) and chronic fatigue syndrome (CFS) are frequently seen in primary care as well as in various medical specialties. Despite a controversy between `lumpers' and `splitters' regarding these syndromes, many authors assume that FM and CFS patients show more communalities than differences. Stress system dysfunctioning and associated abnormal pain processing seem to link these syndromes from a pathophysiological point of view. Consequently, we propose to rename FM and CFS as `stress intolerance and pain hypersensitivity (SIPH) syndromes'. Furthermore, we make a plea for developing specific treatment settings for SIPH patients. Finally, we outline future research perspectives on the interaction between life stress, personality/lifestyle factors, and stress system/pain processing disturbances in the aetio-pathogenesis of SIPH and other functional somatic syndromes.
ABSTRACT Cobalamin (vitamin B12) in the form of hydroxocobalamin or cyanocobalamin injections has been widely used to treat chronic fatigue syndrome (CFS). Hydroxocobalamin is a nitric oxide scavenger and is proposed here to act as such a scavenger in CFS treatment. Its possible efficacy in CFS treatment, if further substantiated, may provide confirmation of a prediction of the elevated nitric oxide/peroxynitrite theory of CFS etiology. This interpretation of the possible role of cobalamin in CFS treatment suggests a useful perspective for confirming and optimizing this treatment.
The radicophilicity (antiradical–antioxidant effects) of processed shilajit (SJP) to oxygen-derived free radicals and nitric oxide (NO), and the attendant H2O2 cleaving effect were evaluated. SJP provided complete protection to methyl methacrylate (MMA) against hydroxyl radical-induced polymerization and acted as a reversible NO-captodative agent. SJP (20 and 50 mg/kg/day, i.p., for 21 days) induced a dose-related increase in superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities in frontal cortex and striatum of rats. The data were comparable to those of (−)-deprenyl (2 mg/kg/day, i.p., for 21 days) in respect of SOD and CAT activities. These findings are consistent with the therapeutic uses of shilajit as an Ayurvedic rasayan (rejuvenator) against oxidative stress and geriatric complaints.
We examined brain region-specific changes in monoamines and metabolites, and their ratios, after short-term administration of antidepressants to rats. Serotonin noradrenaline reuptake inhibitors (SNRIs; duloxetine, venlafaxine, milnacipran) and a serotonin-selective reuptake inhibitor (SSRI; sertraline) elevated serotonin (5-HT) levels in the midbrain (MB). Duloxetine and venlafaxine increased 5-HT levels in the brainstem and 5-HT terminal areas, whereas milnacipran and sertraline increased levels in the brainstem only. Significant reductions in 5-HT turnover were observed in various forebrain regions, including the hippocampus and hypothalamus, after treatment with all of the tested drugs except for milnacipran. In addition, there was reduced 5-HT turnover in the dorsolateral frontal cortex (dlFC), the medial prefrontal cortex (mPFC), and both the dlFC and the mPFC after treatment with duloxetine, sertraline, and venlafaxine, respectively. Venlafaxine significantly increased dopamine (DA) levels in the nucleus accumbens (NAc) and the substantia nigra and decreased DA turnover in the NAc. Similar changes were observed after treatment with duloxetine and sertraline in the NAc, whereas milnacipran increased DA levels in the mPFC. Limited increases in noradrenaline levels were detected after treatment with duloxetine, venlafaxine, or sertraline, but not after treatment with milnacipran. These results show that SNRIs and SSRIs induced region-specific monoaminergic changes after short-term treatment.
We have shown that skin fibroblast from patients with cystic fibrosis (CF) and from carriers for CF [heterozygotes (HZ)] consume more O2 than do their controls. When the mitochondrial electron transport inhibitor rotenone was added to the cells, the relative inhibition of O2 consumption was CF greater than HZ greater than controls (P less than 0.005 in both comparisons). Because rotenone specifically inhibits NADH dehydrogenase, [NADH: (acceptor) oxidoreductase, EC 1.6.99.3], which is the enzyme of energy-conserving site 1 of the mitochondrial electron transport system, activity and kinetics of this enzyme system were studied in fibroblast homogenates. NADH dehydrogenase activity was equal in cells from the three genotypes. At pH 8.0, affinity of the enzyme for its substrate was CF greater than HZ = controls; at pH 8.6, affinity was CF greater than HZ = controls (P less than 0.005 for the differences). pH optima for the genotypes were without exception 8.6 (CF), 8.3 (HZ), and 8.0 (control). HZ and control lines were distinguished unequivocally in a blind test on the basis of differences in pH optima. Purified mitochondrial preparations revealed pH optima identical to those found in whole cell homogenates. These data suggest that the mutant gene responsible for CF is expressed in the complex mitochondrial NADH dehydrogenase system.
Mind and body remain stubbornly one. The distinction between primary and secondary disorders respects this unity. The distinction between "reactive" and "induced" carry causal implications and suggest the former is psychogenic and the latter organic--both of which are probably premature conclusions. The diagnostician, free of the demands on the pathologist, can pursue the correct nosology committed to demonstrating, not the pathophysiology, but the presence of adequate diagnostic criteria. Whenever a secondary disorder meets full criteria it may warrant the same treatment accorded to the primary disorder. Whether the disease is major or minor may also be of clinical significance. Only further application of psychiatric nosology to medically ill patients can resolve these issues. Karajgi et al recently found that the lifetime prevalence of panic disorder in a sample of patients with chronic obstructive pulmonary disease was 8%. The only respectable offspring of neurotic depression in DSM-III-R is dysthymia. As with neurotic depression, dysthymia is not a condition thought appropriate for or responsive to antidepressant drugs. Clinicians dealing with depression in the medically ill think of depression itself as "serious," that is, major.
Microphotometric assay media for the measurement of succinate dehydrogenase (SDH) and cytochrome oxidase activities in sections of human skeletal muscle have been developed. The optimal constitution of these media was determined experimentally. Factors investigated include the effects of substrate concentration, pH, use of different electron acceptors and electron donors, influence of intermediate electron carriers and tissue-stabilizing agents, effects of inhibitors, the extent of endogenous and non-specific reactions and the linearity of the reactions during the time course of the assays. Optimal assay media (SDH) contained 130 mM succinate, 1.5 mM Nitro Blue tetrazolium, 0.2 mM phenazine methosulphate and 1.0 mM sodium azide in 0.1 m phosphate buffer, pH7.0. Cytochrome oxidase was optimally assayed in media containing 4 mM diaminobenzidine and 100 microns cytochrome c. Reactions in individual muscle fibers were found to be linear for incubation times up to 10 min in SDH assays and for more than 15 min in cytochrome oxidase determinations. Some potential uses of these microphotometric assays in the investigation of human metabolic muscle disorders are discussed.
Defects in Complex I of the mitochondrial respiratory chain have been identified in 38 patients. The clinical and laboratory features are reviewed and the results of recently devised strategies aimed at characterizing the primary molecular and genetic abnormalities are presented. Although not exhaustive, these studies have provided a molecular basis for the contention that defects in Complex I may have their origin in nuclear or in mitochondrial genes.
Parenteral administration of nickel chloride (NiCl2) to rats enhanced lipid peroxidation in liver, kidney, and lung (but not in brain, heart, spleen, or testis), as measured by the thiobarbituric acid reaction for malondialdehyde (MDA) and related chromogens in fresh tissue homogenates. After sc injection of NiCl2 (0.75 mmol per kg body wt), MDA concentrations in liver and kidney became significantly increased by nine h and reached peak values at 48 h. For example, in nine rats killed 48 h after the NiCl2 injection, hepatic MDA concentrations averaged 2.5 +/- 1.0 mumol per g dry wt (P less than 0.001 versus 0.5 +/- 0.3 mumol per g in 30 controls). Dose-effect relationships for lipid peroxidation in liver and kidney were observed with NiCl2 dosages ranging from 0.12 to 0.75 mmol per kg, sc. Intrarenal administration of a carcinogenic nickel compound, nickel subsulfide (Ni3S2, 0.36 mmol per kg body wt), did not affect MDA concentrations in the injected kidneys of rats killed one to 20 days post-injection. The results of this study implicate lipid peroxidation as a molecular mechanism for cell injury in acute NiCl2 poisoning, but they do not furnish any evidence that lipid peroxidation is involved in the initiation of nickel carcinogenesis.
Oligomycin resistance is expressed at the mitochondrial and submitochondrial levels in oligomycin‐resistant mutants of Saccharomyces cerevisiae. Studies of the mitochondrial ATPase and P i ‐ATP exchange reaction indicate that a modification of the mitochondrial ATPase (ATP‐synthetase) complex is involved. The specific resistance at the mitochondrial level to oligomycin and related inhibitors such as ossamycin, rutamycin and peliomycin and the lack of resistance to other inhibitors of mitochondrial ATP synthetase (triethyltin sulphate, venturicidin, aurovertin, Dio‐9) correlate with whole cell studies and provide further evidence for a specific modification of an inhibitor site on the mitochondrial inner membrane.
Mitochondrial mutants which map at the OLI and OLII loci can be differentiated biochemically by their sensitivity to oligomycin of mitochondrial ATPase and the pH 9.5/pH 6.5 ATPase activity ratios.
Studies utilising F 1 ‐ATPase from wild type and oligomycin‐resistant mutants indicate that oligomycin resistance is due to a modification of a membrane‐bound component of the mitochondrial ATP‐synthetase complex and not to a modification of F 1 ‐ATPase or the oligomycin‐sensitivity‐conferring protein.
Partially purified oligomycin‐sensitive ATPase preparations (Triton X‐100 extracts) retain the sensitivity (resistance) and specificity of the original membrane‐bound system in mitochondria. Purified oligomycin‐sensitive ATPase preparations also retain the specificity of the original membrane preparation but the sensitivity to inhibitors is partially modified. The results support the conclusion that oligomycin resistance is due to modification of mitochondrially coded and mitochondrially synthesised subunits of the oligomycin‐sensitive ATPase (ATP synthetase) complex. There is no correlation of oligomycin resistance with the ergosterol concentration of yeast cells, yeast mitochondria or mitochondrial ATPase.
Although the mechanism of neuronal death in neurodegenerative diseases remains unknown, it has been hypothesized that relatively minor metabolic defects may predispose neurons to N-methyl-D-aspartate (NMDA) receptor-mediated excitotoxic damage in these disorders. To further investigate this possibility, we have characterized the excitotoxic potential of the reversible succinate dehydrogenase (SDH) inhibitor malonate. After its intrastriatal stereotaxic injection into male Sprague-Dawley rats, malonate produced a dose-dependent lesion when assessed 3 days after surgery using cytochrome oxidase histochemistry. This lesion was attenuated by coadministration of excess succinate, indicating that it was caused by specific inhibition of SDH. The lesion was also prevented by administration of the noncompetitive NMDA antagonist MK-801. MK-801 did not induce hypothermia, and hypothermia itself was not neuroprotective, suggesting that the neuroprotective effect of MK-801 was due to blockade of the NMDA receptor ion channel and not to any nonspecific effect. The competitive NMDA antagonist LY274614 and the glycine site antagonist 7-chlorokynurenate also profoundly attenuated malonate neurotoxicity, further indicating an NMDA receptor-mediated event. Finally, the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) antagonist NBQX (2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)-quinoxaline) was ineffective at preventing malonate toxicity at a dose that effectively reduced S-AMPA toxicity, indicating that non-NMDA receptors are involved minimally, if at all, in the production of the malonate lesion. We conclude that inhibition of SDH by malonate results in NMDA receptor-mediated excitotoxic neuronal death. If this mechanism of "secondary" or "weak" excitotoxicity plays a role in neurodegenerative disease, NMDA antagonists and other "antiexcitotoxic" strategies may have therapeutic potential for these diseases.
Nitric oxide (NO) reversibly inhibited oxygen consumption of brain synaptosomes. Inhibition was reversible, occurred at the level of cytochrome oxidase, and was apparently competitive with oxygen, with half-inhibition by 270 nM NO at oxygen concentrations around 145 microM and by 60 nM at around 30 microM O2. Isolated cytochrome oxidase was inhibited by similar levels of NO. These levels of NO are within the measured physiological and pathological range for a number of tissues and conditions, suggesting that NO inhibition of cytochrome oxidase and the competition with oxygen may occur in vivo.
The complexities of the chronic fatigue syndrome and the methodologic problems associated with its study indicate the need for a comprehensive, systematic, and integrated approach to the evaluation, classification, and study of persons with this condition and other fatiguing illnesses. We propose a conceptual framework and a set of guidelines that provide such an approach. Our guidelines include recommendations for the clinical evaluation of fatigued persons, a revised case definition of the chronic fatigue syndrome, and a strategy for subgrouping fatigued persons in formal investigations.
Hypothalamic-pituitary-adrenal (HPA) axis and central 5-HT function were compared in chronic fatigue syndrome (CFS), depression and healthy states. 10 patients with CFS and 15 patients with major depression were matched for age, weight, sex and menstrual cycle with 25 healthy controls. Baseline-circulating cortisol levels were highest in the depressed, lowest in the CFS and intermediate between the two in the control group (P = 0.01). Prolactin responses to the selective 5-HT-releasing agent d-fenfluramine were lowest in the depressed, highest in the CFS and intermediate between both in the healthy group (P = 0.01). Matched pair analysis confirmed higher prolactin responses in CFS patients than controls (P = 0.05) and lower responses in depressed patients than controls (P = 0.003). There were strong inverse correlations between prolactin and cortisol responses and baseline cortisol values. These data confirm that depression is associated with hypercotisolaemia and reduced central 5-HT neurotransmission and suggest that CFS may be associated with hypocortisolaemia and increased 5-HT function. The opposing responses in CFS and depression may be related to reversed patterns of behavioural dysfunction seen in these conditions. These findings attest to biological distinctions between these disorders.
Recent research employing the elevated plus-maze to assess anxiety in rodents has incorporated a variety of behavioral elements in addition to the standard parameters of entries onto and time spent in the aversive open arms. In the present study, we have used a large database comprising the behavioral profiles of 90 undrugged mice to examine the relationship between the standard spatiotemporal measures and a range of specific behaviors related to the defensive repertoire of the mouse. A factor analysis applied to the standard measures revealed two factors related to anxiety and locomotor activity. The simple addition of center time (an infrequently recorded measure) to the analysis yielded a third factor, most probably related to decision making. A large-scale factor analysis applied to all measures further confirmed the existence of factors related to anxiety, locomotor activity, and decision making, and revealed three further factors thought to represent risk assessment, vertical activity, and exploratory behavior. Thus, the inclusion of ethological measures not only confirmed prior knowledge based on a very limited range of measures, but also demonstrated the existence of additional behavioral dimensions. The potential applications of this knowledge are discussed.
Hypercortisolism in depression seems to preferentially reflect activation of hypothalamic CRH secretion. Although it has been postulated that this hypercortisolism is an epiphenomenon of the pain and stress of major depression, our data showing preferential participation of AVP in the hypercortisolism of chronic inflammatory disease suggest specificity for the pathophysiology of hypercortisolism in depression. Our findings that imipramine causes a down-regulation of the HPA axis in experimental animals and healthy controls support an intrinsic role for CRH in the pathophysiology of melancholia and in the mechanism of action of psychotropic agents. Our data suggest that hypercortisolism is not the only form of HPA dysregulation in major depression. In a series of studies, commencing in patients with Cushing's disease, and extending to hyperimmune fatigue states such as chronic fatigue syndrome and examples of atypical depression such as seasonal affective disorder, we have advanced data suggesting hypofunction of hypothalamic CRH neurons. These data raise the question that the hyperphagia, hypersomnia, and fatigue associated with syndromes of atypical depression could reflect a central deficiency of a potent arousal-producing anorexogenic neuropeptide. In the light of data presented elsewhere in this symposium regarding the role of a hypofunctioning hypothalamic CRH neuron in susceptibility to inflammatory disease, these data also raise the question of a common pathophysiological mechanism in syndromes associated both with inflammatory manifestations and atypical depressive symptoms. This concept of hypofunctioning of hypothalamic CRH neurons in these disorders also raises the question of novel forms of neuropharmacological intervention in both inflammatory diseases and atypical depressive syndromes.
In Experiment 1, rats were tested in a plus-maze, with or without small ledges on the open arms, after injection with vehicle or chlordiazepoxide (7.5 mg/kg). They were scored either on their first or second exposure to the maze; those scored on trail 2 had received a 5-min undrugged exposure to the maze 24 h earlier. This dose of chlordiazepoxide had a significant anxiolytic effect on trial 1 only in the maze without ledges, and on trial 2 only in the maze with ledges; thus, the presence of ledges differentially affected anxiolytic sensitivity on trials 1 and 2. The results of a factor analysis study (Experiment 2) confirmed that ledges had a differential effect when rats were repeatedly exposed to the maze. Thus, in the maze without ledges, the scores reflecting anxiolytic activity on trial 1 loaded on one factor, whereas the scores from trials 2 and 3 loaded on another independent factor. In the maze with ledges, the scores reflecting anxiolytic activity on trials 1, 2, and 3 loaded on three independent factors. Considering the published evidence and the results of the present study, we suggest that both types of plus-maze may be measuring the same type of anxiety with different sensitivities on trial 1 (e.g., generalised anxiety or fear of open spaces); different types of anxiety on trial 2 (without ledges--phobia/fear of heights; with ledges--not known), and trial 3 in the maze with ledges, yet another type of anxiety. The factor analysis results are also presented for ethological measures on the plus-maze, and for activity and exploration in the holeboard. Based on the factor loadings, a composite measure of anxiety on trial 1 is presented which will increase the sensitivity of the plus-maze to anxiolytic treatments. The measures of motor activity in the plus-maze load on a different factor from those derived from the holeboard, thus cautioning against considering all measures of motor activity as interchangeable.
Within the CNS and under normal conditions, nitric oxide (.NO) appears to be an important physiological signalling molecule. Its ability to increase cyclic GMP concentration suggests that .NO is implicated in the regulation of important metabolic pathways in the brain. Under certain circumstances .NO synthesis may be excessive and .NO may become neurotoxic. Excessive glutamate-receptor stimulation may lead to neuronal death through a mechanism implicating synthesis of both .NO and superoxide (O2.-) and hence peroxynitrite (ONOO-) formation. In response to lipopolysaccharide and cytokines, glial cells may also be induced to synthesize large amounts of .NO, which may be deleterious to the neighbouring neurones and oligodendrocytes. The precise mechanism of .NO neurotoxicity is not fully understood. One possibility is that it may involve neuronal energy deficiency. This may occur by ONOO- interfering with key enzymes of the tricarboxylic acid cycle, the mitochondrial respiratory chain, mitochondrial calcium metabolism, or DNA damage with subsequent activation of the energy-consuming pathway involving poly(ADP-ribose) synthetase. Possible mechanisms whereby ONOO- impairs the mitochondrial respiratory chain and the relevance for neurotoxicity are discussed. The intracellular content of reduced glutathione also appears important in determining the sensitivity of cells to ONOO- production. It is concluded that neurotoxicity elicited by excessive .NO production may be mediated by mitochondrial dysfunction leading to an energy deficiency state.
The purpose of this study was to describe the sequence of psychosocial events and infections preceding the onset of chronic fatigue syndrome (CFS). This information was related to the temporal development of crucial symptoms in relation to the onset of, namely, fatigue, sadness, irritability, pain, and feeling of fever.
A personal interview was conducted in 46 patients (mean age, 39.5 years; SD, 9 years) who fulfilled international CFS criteria. These patients were matched with regard to age and gender to 46 carefully matched control subjects. Twenty-three percent of the study subjects were men, and 77% were women. The patient at first identified the month that coincided with the onset of CFS. Similarly, each control subject was asked to identify a "very difficult period" within approximately the same period as the patient with whom the control subject was matched. A list of 14 different life events was perused. Participants were asked to identify for each month whether each of the listed events had occurred. Furthermore, they were asked to rate the importance of the events they had experienced. In addition, for each of the cardinal symptoms (fatigue, sadness, irritability, pain, and feeling of fever) and for each month, the subjects were asked to rate, on a visual analogue scale, the symptom intensity. Also, the number of infections was noted.
A statistically significant group difference in fatigue intensity existed during the period 4 to 10 months before the onset of CFS. During the 3 months preceding the diagnosis for the CFS patients or the peak of the crisis for the control group, there was a dramatic rise in fatigue in both groups. The CFS group reached a much higher fatigue level, which leveled off somewhat during the first year of follow-up but still remained very high in comparison with the control group, which reached precrisis levels 4 months after the peak. Similar patterns were observed for fever and pain. With regard to sadness and irritability, no group difference was observed during the period preceding the crisis. In the patient group, the level stayed high throughout the whole first year of follow-up, whereas a slow return started in the control group; precrisis levels were reached after 1 year in this group. The prevalence ratio (CFS patients/control subjects) for negative events was around 1.0 for the periods 4 to 12 months preceding CFS but 1.9 during the quarter year preceding the onset. For infections, the prevalence ratio increased successively during the four quarters preceding CFS (from 1.4 to 2.3).
According to the retrospective self-reports, there were differences between the groups in fatigue, pain, and feeling of fever during the months preceding the crisis. With regard to depressive and irritable feelings, no preillness differences were reported between the groups. There was a reported excess prevalence of both infections and negative life events during the quarter year preceding the onset of CFS or crisis. Potential sources of error are discussed. These findings must be replicated in longitudinal studies.
The etiology of chronic fatigue syndrome (CFS) has been both obscure and highly contentious, leading to substantial barriers to both clear diagnosis and effective treatment. I propose here a novel hypothesis of CFS in which either viral or bacterial infection induces one or more cytokines, IL-1beta IL-6, TNF-alpha and IFN-gamma. These induce nitric oxide synthase (iNOS), leading to increased nitric oxide levels. Nitric oxide, in turn, reacts with superoxide radical to generate the potent oxidant peroxynitrite. Multiple amplification and positive feedback mechanisms are proposed by which once peroxynitrite levels are elevated, they tend to be sustained at a high level. This proposed mechanism may lower the HPA axis activity and be maintained by consequent lowered glucocorticoid levels. Similarities are discussed among CFS and autoimmune and other diseases previously shown to be associated with elevated peroxynitrite. Multiple pharmacological approaches to the treatment of CFS are suggested by this hypothesis.
The metabolic control of oxidative phosphorylation (OXPHOS) has attracted increasing attention in recent years, especially due to its importance for understanding the role of mitochondrial DNA mutations in human diseases and aging. Experiments on isolated mitochondria have indicated that a relatively small fraction of each of several components of the electron transport chain is sufficient to sustain a normal respiration rate. These experiments, however, may have not reflected the in vivo situation, due to the possible loss of essential metabolites during organelle isolation and the disruption of the normal interactions of mitochondria with the cytoskeleton, which may be important for the channeling of respiratory substrate to the organelles. To obtain direct evidence on this question, in particular, as concerns the in vivo control of respiration by cytochrome c oxidase (COX), we have developed an approach for measuring COX activity in intact cells, by means of cyanide titration, either as an isolated step or as a respiratory chain-integrated step. The method has been applied to a variety of human cell types, including wild-type and mtDNA mutation-carrying cells, several tumor-derived semidifferentiated cell lines, as well as specialized cells removed from the organism. The results obtained strongly support the following conclusions: (i) the in vivo control of respiration by COX is much tighter than has been generally assumed on the basis of experiments carried out on isolated mitochondria; (ii) COX thresholds depend on the respiratory fluxes under which they are measured; and (iii) measurements of relative enzyme capacities are needed for understanding the role of mitochondrial respiratory complexes in human physiopathology.
Mitochondrial damage may play a key role in the development of necrotic and apoptotic hypoxic-ischemic (HI) brain damage. It has previously been shown that mitochondrial respiration is depressed in the cerebral cortex after HI in neonatal animals. The aim of the present study was to further characterize the time course of the mitochondrial impairment during reperfusion and the correlation between the respiratory control ratio and brain injury and activation of caspase-3. Rat pups were subjected to unilateral carotid artery ligation and exposed to hypoxia (7.7% oxygen). Mitochondrial respiration was measured 0-72 h after HI in a mitochondrial fraction isolated from cerebral cortex. Microtubule associated protein-2 (MAP2) and caspase-3 were analyzed with immunoblotting in cerebral cortex homogenates. In addition, the time course of caspase-3 activation was measured as DEVD cleavage. The mitochondrial respiratory control ratio in cerebral cortex decreased immediately after HI followed by a partial recovery at 3-8 h. Thereafter, a secondary drop occurred with a minimum reached at 24 h of reperfusion. The secondary loss of respiratory function was accompanied by depletion of MAP2, cleavage of caspase-3 and an increased caspase-3 -like activity at 3-24 h after the insult. In conclusion, the primary phase of mitochondrial dysfunction was paralleled by a moderate decrease of MAP2 and a limited activation of caspase-3. The secondary mitochondrial impairment was associated with neuronal injury and pronounced activation of caspase-3.
Generalized anxiety disorder (GAD) is a common and serious disorder. Despite this fact, there is no clear understanding of the exact neurobiological changes underlying the condition. To date, there are few studies of neurobiological function in patients with GAD, and only limited comparative data with depression are available. Advances in neuroanatomical imaging techniques are beginning to allow detailed study of regional blood flow and metabolism and may offer insights into the specific regions of the brain involved in GAD. Investigations into neurotransmitter dysfunction have implicated the gamma-aminobutyric acid/benzodiazepine, serotonergic, and noradrenergic systems in this disorder. Variations in sleep patterns have also been assessed and indicate a biological separation from depression.