Nagwa Sayed Ahmed

Confirmatory studies on the antioxidant and antidiabetic effect of quercetin in rats

Quercetin (QE), one of natural flavanoid group, was widely distributed as a secondary metabolite in plant kingdom. It has been believed that oxidative stress plays a role in the pathogenesis of diabetes mellitus (DM). The aim of the present study was the evaluation of possible effects of QE on blood glucose and antioxidant enzymes in experimental streptozotocin (STZ)-induced diabetes in rats. STZ was injected intraperitoneally with single dose of 50 mg/kg for diabetes induction. QE (15 mg/kg bw day, intraperitoneal (i.p.) injection) was injected for 3 days prior to STZ administration; these injections were continued to the end of the study (for 25 days). Glucose tolerance test and random plasma glucose were done for all animals. Cellular antioxidant enzymes such as glutathione peroxidase (GSHPx), superoxide dismutase (SOD) and catalase (CAT) activities were measured in pancreatic homogenates. Quercetin had no effect on plasma glucose level of normal animals but its pre- treatment was able to prevent diabetes induced by single intraperitoneal injection of streptozocintreated rats. Antioxidant enzyme activity significantly decreased in STZ induced diabetic group. QE treatment significantly increased the antioxidant enzyme activities. It could be concluded that quercetin, a flavonoid with antioxidant properties, exerting its beneficial antidiabetic effects.

Safety and outcome of treatment of metastatic melanoma using 3-bromopyruvate: a concise literature review and case study

3-Bromopyruvate (3BP) is a new, promising anticancer alkylating agent with several notable functions. In addition to inhibiting key glycolysis enzymes including hexokinase II and lactate dehydrogenase (LDH), 3BP also selectively inhibits mitochondrial oxidative phosphorylation, angiogenesis, and energy production in cancer cells. Moreover, 3BP induces hydrogen peroxide generation in cancer cells (oxidative stress effect) and competes with the LDH substrates pyruvate and lactate. There is only one published human clinical study showing that 3BP was effective in treating fibrolamellar hepatocellular carcinoma. LDH is a good measure for tumor evaluation and predicts the outcome of treatment better than the presence of a residual tumor mass. According to the Warburg effect, LDH is responsible for lactate synthesis, which facilitates cancer cell survival, progression, aggressiveness, metastasis, and angiogenesis. Lactate produced through LDH activity fuels aerobic cell populations inside tumors via metabolic symbiosis. In melanoma, the most deadly skin cancer, 3BP induced necrotic cell death in sensitive cells, whereas high glutathione (GSH) content made other melanoma cells resistant to 3BP. Concurrent use of a GSH depletor with 3BP killed resistant melanoma cells. Survival of melanoma patients was inversely associated with high serum LDH levels, which was reported to be highly predictive of melanoma treatment in randomized clinical trials. Here, we report a 28-year-old man presented with stage IV metastatic melanoma affecting the back, left pleura, and lung. The disease caused total destruction of the left lung and a high serum LDH level (4,283 U/L). After ethics committee approval and written patient consent, the patient received 3BP intravenous infusions (1-2.2 mg/kg), but the anticancer effect was minimal as indicated by a high serum LDH level. This may have been due to high tumor GSH content. On combining oral paracetamol, which depletes tumor GSH, with 3BP treatment, serum LDH level dropped maximally. Although a slow intravenous infusion of 3BP appeared to have minimal cytotoxicity, its anticancer efficacy via this delivery method was low. This was possibly due to high tumor GSH content, which was increased after concurrent use of the GSH depletor paracetamol. If the anticancer effectiveness of 3BP is less than expected, the combination with paracetamol may be needed to sensitize cancer cells to 3BP-induced effects.

Warburg effect increases steady-state ROS condition in cancer cells through decreasing their antioxidant capacities (Anticancer effects of 3-bromopyruvate through antagonizing Warburg effect)

Cancer cells undergo an increased steady-state ROS condition compared to normal cells. Among the major metabolic differences between cancer cells and normal cells is the dependence of cancer cells on glycolysis as a major source of energy even in the presence of oxygen (Warburg effect). In Warburg effect, glucose is catabolized to lactate that is extruded through monocarboxylate transporters to the microenvironment of cancer cells, while in normal cells, glucose is metabolized into pyruvate that is not extruded. Pyruvate is a potent antioxidant, while lactate has no antioxidant effect. Pyruvate in normal cells may be further metabolized to acetyl CoA and then through Krebs cycle with production of antioxidant intermediates e.g. citrate, malate and oxaloacetate together with the reducing equivalents (NADH.H+). Through activity of mitochondrial transhydrogenase, NADH.H+ replenishes NADPH.H+, coenzyme of glutathione reductase which replenishes reduced form of glutathione (potent antioxidant). This enhances antioxidant capacities of normal cells, while cancer cells exhibiting Warburg effect may be deprived of all that antioxidant capabilities due to loss of extruded lactate (substrate for Krebs cycle). Although intrinsic oxidative stress in cancer cells is high, it may be prevented from reaching progressively increasing levels that are cytotoxic to cancer cells. This may be due to some antioxidant effects exerted by hexokinase II (HK II) and NADPH.H+ produced through HMP shunt. Glycolytic phenotype in cancer cells maintains a high non-toxic oxidative stress in cancer cells and may be responsible for their malignant behavior. Through HK II, glycolysis fuels the energetic arm of malignancy, the mitotic arm of malignancy (DNA synthesis through HMP shunt pathway) and the metastatic arm of malignancy (hyaluronan synthesis through uronic acid pathway) in addition to the role of phosphohexose isomerase (autocrine motility factor). All those critical three arms start with the substrate G6P that is a direct product of HK II. 3-bromopyruvate (3BP, inhibitor of HK II) may prove as a promising anticancer and antimetastatic agent based on antagonizing the Warburg effect and disturbing the malignant behavior in cancer cells.

Curcumin ameliorate DENA-induced HCC via modulating TGF-β, AKT, and caspase-3 expression in experimental rat model

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. In laboratory animal models, diethylnitrosamine (DENA) is a well-known agent that has a potent hepatocarcinogenic effect that is used to induce HCC. As curcumin has a potent anti-inflammatory effect with strong therapeutic potential against a variety of cancers, our present study aims to investigate its curative effects and the possible mechanisms of action against DENA-induced HCC in male rats. Investigation of biochemical and molecular parameters of HCC animal model liver showed an overexpression of TGF-β and Akt proteins accompanied with a significant reduction of the proapoptotic marker caspase-3. DENA-induced hepatic cellular injury resulted also in a significant increase in liver function marker enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lipid peroxides in this group. Curcumin treatment partially reversed DENA-induced damage as it reduced the overexpression of the angiogenic and anti-apoptotic factors TGF-β and Akt and improved caspase-3 expression. Also, it could partially normalize the serum values of liver marker enzymes and lipid peroxidation and improve liver architecture. Curcumin shows a unique chemotherapeutic effect in reversing DENA-induced HCC in rat model. This effect is possibly mediated through its proapoptotic, antioxidant, anti-angiogenic, as well as antimitotic effects. It interferes and modulates cell signaling pathways and hence turns death signals and apoptosis on within tumor cells.

Percutaneous excretion of iron and ferritin (through Al-hijamah) as a novel treatment for iron overload in beta-thalassemia major, hemochromatosis and sideroblastic anemia

Iron overload is a big challenge when treating thalassemia (TM), hemochromatosis and sideroblastic anemia. It persists even after cure of TM with bone marrow transplantation. Iron overload results from increased iron absorption and repeated blood transfusions causing increased iron in plasma and interstitial fluids. Iron deposition in tissues e.g. heart, liver, endocrine glands and others leads to tissue damage and organ dysfunction. Iron chelation therapy and phlebotomy for iron overload have treatment difficulties, side effects and contraindications. As mean iron level in skin of TM patients increases by more than 200%, percutaneous iron excretion may be beneficial. Wet cupping therapy (WCT) is a simple, safe and economic treatment. WCT is a familiar treatment modality in some European countries and in Chinese hospitals in treating different diseases. WCT was reported to clear both blood plasma and interstitial spaces from causative pathological substances (CPS). Standard WCT method is Al-hijamah (cupping, puncturing and cupping, CPC) method of WCT that was reported to clear blood and interstitial fluids better than the traditional WCT (puncturing and cupping method, PC method of WCT). In other word, traditional WCT may be described as scarification and suction method (double S technique), while Al-hijamah may be described as suction, scarification and suction method (triple S technique). Al-hijamah is a more comprehensive treatment modality that includes all steps and therapeutic benefits of traditional dry cupping therapy and WCT altogether according to the evidence-based Taibah mechanism (Taibah theory). During the first cupping step of Al-hijamah, a fluid mixture is collected inside skin uplifting due to the effect of negative pressure inside sucking cups. This fluid mixture contains collected interstitial fluids with CPS (iron, ferritin and hemolyzed RBCs in thalassemia), filtered fluids (from blood capillaries) with iron and hemolyzed blood cells (hemolyzed RBCs, WBCs and platelets). That fluid mixture does not contain intact blood cells (having diameters in microns) that are too big to pass through pores of skin capillaries (6-12nm in diameter) and cannot be filtered. Puncturing skin upliftings and applying second cupping step excrete collected fluids. Skin scarifications (shartat mihjam in Arabic) should be small, superficial (0.1mm in depth), short (1-2mm in length), multiple, evenly distributed and confined to skin upliftings. Sucking pressure inside cups (-150 to -420mmHg) applied to skin is transmitted to around skin capillaries to be added to capillary hydrostatic pressure (-33mmHg at arterial end of capillaries and -13mmHg at venous end of capillaries) against capillary osmotic pressure (+20mmHg). This creates a pressure gradient and a traction force across skin and capillaries and increases filtration at arterial end of capillaries at net pressure of -163 to -433mmHg and at venous end of capillaries at net pressure of -143 to -413mmHg resulting in clearance of blood from CPS (iron, ferritin and hemolyzed blood cells). Net filtration pressure at renal glomeruli is 10mmHg i.e. Al-hijamah exerts a more pressure-dependent filtration than renal glomeruli. Al-hijamah may benefit patients through inducing negative iron balance. Interestingly, Al-hijamah was reported to decrease serum ferritin significantly (by about 22%) in healthy subjects while excessive traditional WCT was reported to cause iron deficiency anemia. Al-hijamah is a highly recommended treatment in prophetic medicine. In conclusion, Al-hijamah may be a promising adjuvant treatment for iron overload in TM, hemochromatosis and sideroblastic anemia.

Al-hijamah and oral honey for treating thalassemia, conditions of iron overload, and hyperferremia: toward improving the therapeutic outcomes.

Iron overload causes iron deposition and accumulation in the liver, heart, skin, and other tissues resulting in serious tissue damages. Significant blood clearance from iron and ferritin using wet cupping therapy (WCT) has been reported. WCT is an excretory form of treatment that needs more research efforts. WCT is an available, safe, simple, economic, and time-saving outpatient modality of treatment that has no serious side effects. There are no serious limitations or precautions to discontinue WCT. Interestingly, WCT has solid scientific and medical bases (Taibah mechanism) that explain its effectiveness in treating many disease conditions differing in etiology and pathogenesis. WCT utilizes an excretory physiological principle (pressure-dependent excretion) that resembles excretion through renal glomerular filtration and abscess evacuation. WCT exhibits a percutaneous excretory function that clears blood (through fenestrated skin capillaries) and interstitial fluids from pathological substances without adding a metabolic or detoxification burden on the liver and the kidneys. Interestingly, WCT was reported to decrease serum ferritin (circulating iron stores) significantly by about 22.25% in healthy subjects (in one session) and to decrease serum iron significantly to the level of causing iron deficiency (in multiple sessions). WCT was reported to clear blood significantly of triglycerides, low-density lipoprotein (LDL) cholesterol, total cholesterol, uric acid, inflammatory mediators, and immunoglobulin antibodies (rheumatoid factor). Moreover, WCT was reported to enhance the natural immunity, potentiate pharmacological treatments, and to treat many different disease conditions. There are two distinct methods of WCT: traditional WCT and Al-hijamah (WCT of prophetic medicine). Both start and end with skin sterilization. In traditional WCT, there are two steps, skin scarification followed by suction using plastic cups (double S technique); Al-hijamah is a three-step procedure that includes skin suction using cups, scarification (shartat mihjam in Arabic), and second skin suction (triple S technique). Al-hijamah is a more comprehensive technique and does better than traditional WCT, as Al-hijamah includes two pressure-dependent filtration steps versus one step in traditional WCT. Whenever blood plasma is to be cleared of an excess pathological substance, Al-hijamah is indicated. We will discuss here some reported hematological and therapeutic benefits of Al-hijamah, its medical bases, methodologies, precautions, side effects, contraindications, quantitative evaluation, malpractice, combination with oral honey treatment, and to what extent it may be helpful when treating thalassemia and other conditions of iron overload and hyperferremia.

Immunohistochemical Analysis of GDNF and Its Cognate Receptor GFRα-1 Protein Expression in Vitiliginous Skin Lesions.

Background: Vitiligo is an idiopathic skin disease, characterized by circumscribed white macules or patches on the skin due to loss of the functional melanocytes. Glial cell line–derived neurotrophic factor (GDNF) and its cognate receptor (GFRα-1) are distal members of the transforming growth factor-β superfamily. GDNF, produced by the basal cell keratinocytes, is involved in the migration and differentiation of the melanocytes from the neural crest to the epidermis. This study examines the hypothesis that expression of GDNF protein and its cognate receptor GFRα-1 protein is altered in vitiliginous skin. Patients and Methods: To test our hypothesis, we examined the expression patterns of these proteins in vitiliginous and corresponding healthy (control) skin biopsies (20 specimens each) using immunoperoxidase staining techniques. Results: We found variations between the vitiliginous skin and healthy skin. In healthy skin, the expression of GDNF and GFRα-1 proteins was strong (basal cell keratinocytes and melanocytes), moderate (spinous layer), and weak (granular cell layer). In contrast, weak expression of GDNF protein was observed in all epidermal layers of vitiliginous skin. GFRα-1 protein expression was strong (basal cell keratinocytes and melanocytes), moderate (spinous layer), and weak (granular cell layer). In both healthy skin and vitiliginous skin, the expression of GDNF and GFRα-1 proteins was strong in the adnexal structures. Conclusions: We report, for the first time, decreased expression of GDNF proteins in the epidermal keratinocytes of vitiliginous skin. Our findings suggest possible pathogenetic roles for these proteins in the development of vitiligo. The clinical ramifications of these observations mandate further investigations.

Lesional and circulating levels of interleukin-17 and 25-hydroxycholecalciferol in active acne vulgaris: Correlation to disease severity

The immunological aspects of inflammatory acne are still incompletely understood, so this study aimed to investigate the possible role of IL‐17 and 25 hydroxycholecalciferol (25(OH)D3) in the disease pathogenesis and progression.

Lesional levels of superoxide dismutase and malondialdehyde in paucibacillary and multibacillary leprosy patients

Background Leprosy or Hansen’s disease (HD) is a chronic infectious disease caused by Mycobacterium leprae. The ratio of malondialdehyde (MDA)/superoxide dismutase (SOD) may be considered as an index of oxidative stress (OS). Objective To assess the OS by estimating the levels of antioxidant enzyme (SOD), lipid peroxidation products (MDA) and the ratio of MDA/SOD in the skin of paucibacillary (PB) and multibacillary (MB) leprosy patients. Patients and methods This case-control study was conducted on 41 untreated leprosy patients (18 patients with PB type and 23 patients with MB type) attending Sohag’s Dermatology and Leprosy Clinic, in addition to 18 healthy controls. Colorimetric assays of MDA and SOD levels in the tissue homogenates of the skin biopsies taken from the patient and control groups were performed. Results There were statistically significantly higher levels of tissue MDA and statistically significantly lower levels of tissue SOD in leprosy patients when compared with the control group (P<0.001) and in MB leprosy patients when compared with PB leprosy patients (P<0.001). There was statistically significantly higher tissue OS in leprosy patients when compared with the control group (P<0.01) and in MB leprosy patients when compared with PB leprosy patients (P<0.001). There were significant negative correlations between SOD with both MDA and OS with a significant positive correlation between MDA and OS in both patients versus controls and in MB leprosy versus PB leprosy patients (P<0.01 for all). Conclusion OS may play an important role in the pathogenesis of leprosy. The imbalance between oxidant/antioxidant mechanisms may be the contributing factor in the severity of the disease.

Role of Aminoguanidine and Nicorandil in Oxidative Stress in Streptozotocin-Induced Diabetes Mellitus in Rats

Abstract: The effect of aminoguanidine, nicorandil and their combination against oxidative stress in streptozotocin (STZ) induced diabetes mellitus was assessed in rats by determining changes in blood glucose level, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) in healthy and experimentally induced diabetic rats. Besides, histhopathological examination of kidney and liver tissues was performed. Diabetic rats were randomized into groups of six rats and received 50mg/kg, intraperitoneally of aminoguanidine (an antiadvanced glycation end products (AGE) which prevents the formation of reactive oxygen species and lipid peroxidation in cells), 0.1mg/kg, orally of nicorandil (nicotinamide derivative which is efficacious in the treatment of hypertension and angina pectoris and a potassium channel opener) and their combination once daily for one month. Blood glucose level was significantly elevated in plasma of diabetic rats. SOD, CAT and GSH levels were significantly reduced, while MDA and NO levels were significantly elevated in plasma of diabetic rats. Abnormalities in both kidney and liver structures of diabetic rats were observed. Treatments of the diabetic rats with aminoguanidine, nicorandil and their combination led to improvement of the abnormalities in SOD, CAT, GSH, MDA, NO and also the histhopathological abnormalities of kidney and liver. From these results it can be concluded that aminoguanidine, nicorandil and their combination have the ability to attenuate oxidative stress induced by streptozotocin. This effect is positively correlated with their anti-oxidant activities. The effect of aminoguanidine, nicorandil and their combination against oxidative stress in streptozotocin (STZ) induced diabetes mellitus was assessed in rats by determining changes in blood glucose level, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) in healthy and experimentally induced diabetic rats. Besides, histhopathological examination of kidney and liver tissues was performed. Diabetic rats were randomized into groups of six rats and received 50mg/kg, intraperitoneally of aminoguanidine (an antiadvanced glycation end products (AGE) which prevents the formation of reactive oxygen species and lipid peroxidation in cells), 0.1mg/kg, orally of nicorandil (nicotinamide derivative which is efficacious in the treatment of hypertension and angina pectoris and a potassium channel opener) and their combination once daily for one month. Blood glucose level was significantly elevated in plasma of diabetic rats. SOD, CAT and GSH levels were significantly reduced, while MDA and NO levels were significantly elevated in plasma of diabetic rats. Abnormalities in both kidney and liver structures of diabetic rats were observed. Treatments of the diabetic rats with aminoguanidine, nicorandil and their combination led to improvement of the abnormalities in SOD, CAT, GSH, MDA, NO and also the histhopathological abnormalities of kidney and liver. From these results it can be concluded that aminoguanidine, nicorandil and their combination have the ability to attenuate oxidative stress induced by streptozotocin. This effect is positively correlated with their anti-oxidant activities.