A Low Power Digital IC Design Inside the Wireless Endoscopy Capsule. Xanya Sofra Weiss

Xiang Xie, GuoLin Li, XinKai Chen, Lu Liu, Chun Zhang and ZhiHua Wang

This paper proposes an analog-digital mix-mode low power IC architecture inside the wireless endoscopic capsule, which assures that the capsule can implement the diagnoses of whole human digestive tract and provides real time endoscopic image monitoring. A new low complexity and low power digital IC design inside the wireless endoscopic capsule is discussed in detail. To decrease the power consumption, a novel architecture of three-stage clock management is applied for such a system. A new image compression algorithm based on Bayer image format and its corresponding hardware architecture are both presented for low power and low communication data rate application. The digital circuits were verified on FPGAs and have been implemented in 0.18μm CMOS process.

Xanya Sofra Weiss

Xanya Sofra Weiss

Microcurrent therapeutic technique for treatment of radiation toxicity. Xanya Sofra Weiss

Arlene Lennox, Sandra Funder; 2000

The present technique provides a method of remediating the toxicities associated with radiation therapy. A conductive gel is applied to the affected bodily area. A sinusoidally pulsed biphasic DC current is then applied to the affected bodily area using at least one electrode. The electrode is manipulated using active tactile manipulation by for a predetermined time and the frequency of the sinusoidally pulsed biphasic DC current is decreased during the course of the treatment. The method also includes applying a spiked pulsed biphasic DC current to the affected bodily area using at least one electrode. This electrode is also manipulated using active tactile manipulation by for a predetermined time and the frequency of the spiked pulsed biphasic DC current is also decreased during the course of the treatment.

Xanya Sofra Weiss

Xanya Sofra Weiss

Studies of principle and method to depress the air-gap effect to the clamp-on micro-current transducer. Xanya Sofra Weiss

Kai Zhou Pan Chen Buxin You Hao Yang Yang Li
Jiangbei Power Supply Bur. of Chongqing Electr. Power Corp., Chongqing; 2008

The amplitude-phase characteristic of the transducer and the effect of the gap to the magnetic resistance, inductance and mutual inductance of the transducer are analyzed by the equivalent model of passing through clamp-on micro-current transducer. Because of the gap, the lower limiting frequency of the transducer increase and the bandwidth turn narrow, as well as the voltage induced by the coil of the transducer is so feebleness and easy to be interfered that it is difficult to be measured accurately, which is a well-known difficult problem in the portable type measurement of the low-frequency feeble current signal. In this paper, a new method with design principle and the simulation result by Saber to reduce the effect of the gap in the transducer is introduced. The academic lower limiting frequency of the transducer is nearly zero and the upper limiting frequency is infinity. The test of the current transducer shows that the low limiting frequency of the transducer is improved, which can be use in most systems to reduce the effect of gap with broad pass-bandwidth, high linearity and high sensitivity.

Xanya Sofra Weiss

Xanya Sofra Weiss

A prospective evaluation of insulin and insulin-like growth factor-I as risk factors for endometrial cancer. Xanya Sofra Weiss

Gunter MJ, Hoover DR, Yu H, Wassertheil-Smoller S, Manson JE, Li J ,Harris TG, Rohan TE, Xue X, Ho GY, Einstein MH, Kaplan RC, Burk RD,Wylie-Rosett J, Pollak MN, Anderson G, Howard BV, Strickler HD;  2008

Obesity is a major risk factor for endometrial cancer, a relationship thought to be largely explained by the prevalence of high estrogen levels in obese women. Obesity is also associated with high levels of insulin, a known mitogen. However, no prospective studies have directly assessed whether insulin and/or insulin-like growth factor-I (IGF-I), a related hormone, are associated with endometrial cancer while accounting for estrogen levels. We therefore conducted a case-cohort study of incident endometrial cancer in the Women's Health Initiative Observational Study, a prospective cohort of 93,676 postmenopausal women. The study involved all 250 incident cases and a random subcohort of 465 subjects for comparison. Insulin, total IGF-I, free IGF-I, IGF-binding protein-3, glucose, and estradiol levels were measured in fasting baseline serum specimens. Cox models were used to estimate associations with endometrial cancer, particularly endometrioid adenocarcinomas, the main histologic type (n = 205). Our data showed that insulin levels were positively associated with endometrioid adenocarcinoma [hazard ratio contrasting highest versus lowest quartile (HR(q4-q1)), 2.33; 95% confidence interval (95% CI), 1.13-4.82] among women not using hormone therapy after adjustment for age and estradiol. Free IGF-I was inversely associated with endometrioid adenocarcinoma (HR(q4-q1), 0.53; 95% CI, 0.31-0.90) after adjustment for age, hormone therapy use, and estradiol. Both of these associations were stronger among overweight/obese women, especially the association between insulin and endometrioid adenocarcinoma (HR(q4-q1), 4.30; 95% CI, 1.62-11.43). These data indicate that hyperinsulinemia may represent a risk factor for endometrioid adenocarcinoma that is independent of estradiol. Free IGF-I levels were inversely associated with endometrioid adenocarcinoma, consistent with prior cross-sectional data.

Xanya Sofra Weiss

Xanya Sofra Weiss

Effect of microcurrent electrical tissue stimulation on equine tenocytes in culture. Xanya Sofra Weiss

Yi-lo Lin, DVM, MSc Hugo Moolenaar, PhD P. René van Weeren, DVM, PhD Chris H. A. van de Lest, PhD; 2006

Objective—To determine effects of microcurrent electrical tissue stimulation (METS) on equine tenocytes cultured from the superficial digital flexor tendon (SDFT).

Sample Population—SDFTs were collected from 20 horses at slaughter.

Procedure—Tenocytes were isolated following outgrowth from explants and grown in 48-well plates. Four methods of delivering current to the tenocytes with a METS device were tested. Once the optimal method was selected, current consisting of 0 (negative control), 0.05, 0.1, 0.5, 1.0, or 1.5 mA was applied to cells (8 wells/current intensity) once daily for 8 minutes. Cells were treated for 1, 2, or 3 days. Cell proliferation, DNA content, protein content, and apoptosis rate were determined.

Results—Application of microcurrent of moderate intensity increased cell proliferation and DNA content, with greater increases with multiple versus single application. Application of microcurrent of moderate intensity once or twice increased protein content, but application 3 times decreased protein content. Application of current a single time did not significantly alter apoptosis rate; however, application twice or 3 times resulted in significant increases in apoptosis rate, and there were significant linear (second order) correlations between current intensity and apoptosis rate when current was applied twice or 3 times.

Conclusions and Clinical Relevance—Results of the present study indicate that microcurrent affects the behavior of equine tenocytes in culture, but that effects may be negative or positive depending on current intensity and number of applications. Therefore, results are far from conclusive with respect to the suitability of using METS to promote tendon healing in horses.

Xanya Sofra Weiss

Xanya Sofra Weiss

Exercise and arthritis. The hematology of inactivity.Xanya Sofra Weiss

Arthritis tends to promote inactivity, and inactivity tends to promote an unhealthful constellation of blood abnormalities that increases the risk of heart attack and stroke. The hematology of inactivity comprises the following: low plasma volume, high hematocrit, high plasma fibrinogen, elevated blood viscosity, increased platelet aggregability, and diminished fibrinolysis. Regular exercise reverses all these adverse blood changes and, thereby, helps prevent heart attack and stroke. Simply put, exercise "improves" the blood, making it flow more easily and clot less readily. This "healthy hematology of exercisers" is one more reason why prudent exercise is as vital for patients with arthritis as it is for the rest of us.

Xanya Sofra Weiss

Xanya Sofra Weiss

nduction of microcurrents in critically ill patients in magnetic resonance systems. Xanya Sofra Weiss Xanya Sofra Weiss

Measurements and Main Results: Voltage generated by saline 0.9% flowing through a magnetic field and distribution of current from a catheter tip within a sheep heart model were measured in a 0.15 Tesla MRI system. Resistance of loops formed by pacing wires, a pacing electrode, and a thermistor wire were measured in saline 0.9%. Effects of rapidly changing magnetic fields and the movement of the beating heart on epicardial pacing wires were calculated theoretically. A flow of 200 mL/min of saline 0.9% induced a current of 0.1 microampere (uA) (at 0.15 Tesla). From magnetic resonance images we derived a current density of -0.004 [mu]A/mm2 (at 0.15 Tesla). Internal resistance of pacing catheters and thermistor wires was >1 megaohm (M[OMEGA]). The maximum currents calculated (for a higher field strength of 1.5 Tesla) in a circuit formed by epicardial pacing wires were 80 [mu]A, induced by the beating heart moving the wires through the magnetic field and 46 [mu]A, induced by the rapidly changing magnetic fields.

Conclusions: Current generated by flow of conducting fluid should be safe. Pacing catheters and thermistor wires should be safe if well insulated and disconnected from external electric connections. However, current induced in epicardial pacing wires may be a hazard, and precautions should be taken. External wire tips must be separated, insulated, and coiled to lie along the axis of the magnetic field. Electrocar-diography is required, and defibrillation equipment should be available.

Xanya Sofra Weiss

Xanya Sofra Weiss

APPARATUS FOR PERFORMING MICROCURRENT ELECTROTHERAPY. Xanya Sofra Weiss

An electrotherapy method and apparatus for healing injuries and tissue diseases to the human or animal body is disclosed. Particularly, the electrotherapy method disclosed herein comprises delivering a current under various conditions, such as, applying microcurrents ranging from 4 miliamperes to 1 femtoamperes, applying an laternating current with a frequency in the range of .00065 Hz to .00085 Hz, utilizing large surface area electrodes to achieve to low current densities of less than 5 microamperes per square inch. The apparatus of the present invention includes a plurality of electrode wraps for applying said electrotherapy method to a body. Each electrode wrap includes a first layer wrap of water absorptive material and a second layer wrap of moderately conductive material. Each electrode wrap is placed on a portion of the body, including, without limitation, arms, legs, hands, feet and torso. The method and apparatus of the present invention are useful in treating wounds, ulcerations, spinal cord injuries, amyotrophic lateral sclerosis, multiple sclerosis, nervous system abnormalities, scar tissue and age lines.

Xanya Sofra Weiss

Xanya Sofra Weiss

A redox-sensitive peroxiredoxin that is important for longevity has tissue- and stress-specific roles in stress resistance. Xanya Sofra Weiss

Oxidative damage caused by reactive oxygen species (ROS) is implicated in many diseases and in aging. Removal of ROS by antioxidant enzymes plays an important part in limiting this damage. For instance, peroxiredoxins (Prx) are conserved, abundant, thioredoxin peroxidase enzymes that function as tumor suppressors. In addition to detoxifying peroxides, studies in single-cell systems have revealed that Prx act as chaperones and redox sensors. However, it is unknown in what manner the different activities of Prx influence stress resistance or longevity in the context of whole animals. Here, we reveal three distinct roles for the 2-Cys Prx, PRDX-2, in the stress resistance of the nematode worm Caenorhabditis elegans. (i) The thioredoxin peroxidase activity of PRDX-2 protects against hydrogen peroxide. (ii) Consistent with a chaperone activity for hyperoxidized PRDX-2, peroxide-induced oxidation of PRDX-2 increases resistance to heat stress. (iii) Unexpectedly, loss of PRDX-2 increases the resistance of C. elegans to some oxidative stress-causing agents, such as arsenite, apparently through a signaling mechanism that increases the levels of other antioxidants and phase II detoxification enzymes. Despite their increased resistance to some forms of oxidative stress, prdx-2 mutants are short-lived. Moreover, intestinal expression of PRDX-2 accounts for its role in detoxification of exogenous peroxide, but not its influence on either arsenite resistance or longevity, suggesting that PRDX-2 may promote longevity and protect against environmental stress through different mechanisms. Together the data reveal that in metazoans Prx act through multiple biochemical activities, and have tissue-specific functions in stress resistance and longevity.

Xanya Sofra Weiss

Xanya Sofra Weiss

Melatonin in the skin: synthesis, metabolism and functions. Xanya Sofra Weiss

Andrzej Slominski, Desmond J. Tobin, Michal A. Zmijewski, Jacobo Wortsman and Ralf Paus

Melatonin, a ubiquitous methoxyindole, is produced by and metabolized in the skin. Melatonin affects skin functions and structures through actions mediated by cell-surface and putative-nuclear receptors expressed in skin cells. Melatonin has both receptor-dependent and receptor-independent effects that protect against oxidative stress and can attenuate ultraviolet radiation-induced damage. The widespread expression and pleiotropic activity of the cutaneous melatoninergic system provides for a high level of cell-specific selectivity. Moreover, intra-, auto- and para-crine mechanisms equip this system with exquisite functional selectivity. The properties of endogenous melatonin suggest that this molecule is an important effector of stress responses in the skin. In this way, melatonin
actions may counteract or buffer both environmental and endogenous stressors to maintain skin integrity.

Xanya Sofra Weiss

Xanya Sofra Weiss

Acupuncture: neuropeptide release produced by electrical stimulation of different frequencies. Xanya Sofra Weiss

Brain functions are regulated by chemical messengers that include neurotransmitters and neuropeptides. Recent studies have shown that acupuncture or electrical stimulation in specific frequencies applied to certain body sites can facilitate the release of specific neuropeptides in the CNS, eliciting profound physiological effects and even activating self-healing mechanisms. Investigation of the conditions controlling this neurobiological reaction could have theoretical and clinical

Xanya Sofra Weiss

Xanya Sofra Weiss

Ionic Requirements for PCH-Jnduced Pigment Aggregation in the Freshwater Shrimp, Macrobruchium potiuna, Erythrophores. Xanya Sofra Weiss

Ana Lucia M. Britto, Lars Josefsson, Eliuna Scemes,Maria Aparecida Visconti, and Ana Maria de L. Castrucci

The effects of either cation removal or ionic channel blockade were determined on the doseresponse curve (DRC) to PCH (pigment-concentrating hormone) in Macrobmchium potiuna erythrophores. In sodium-, potassium- and calcium-free salines, the pigment-aggregating responses to PCH were depressed; in the former condition, maximal aggregation was not achieved and the slope of the regression curve determined from the DRC was significantly different from control. Tetrodotoxin, verapamil or tetraethylammonium (TEA) treatments also diminished the pigment-aggregating responses to PCH, and the slopes of the regression curves were different from control in the presence of 10m6 M verapamil or 10m6 M TEA. Interestingly, the DRC determined in the absence of both sodium and calcium ions was not significantly different from control. When verapamil was applied in sodium-free conditions, maximal aggregation was prevented. The erythrophore resting membrane potential ranged from -62 mV to - 78 mV and did not vary during PCH-induced pigment aggregation as compared to the control. Our results suggest that transient modifications of potassium equilibrium potential may interfere with PCH signal transduction, revealing a more relevant role of potassium in the process, and that a sodium influx and an intracellular calcium mobilization are necessary to maintain a cytosolic balance between the ions for normality of PCH-induced responses. (COMP BIOCHEM PHYSIOL 113A;4:351-359, 1996

Xanya Sofra Weiss

Xanya Sofra Weiss

Roles of Exercise and Pharmacokinetics in Cerivastatin-Induced Skeletal Muscle Toxicity

Jennifer L. Seachrist, Cho-Ming Loi, Mark G. Evans, Kay A. Criswell, and Charles E. Rothwell; 2005

Three-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are associated with adverse skeletal muscle effects, but the underlying mechanisms remain unclear. To determine whether toxicity involves the level of drug exposure in muscle tissue and to test the effect of exercise on cerivastatin (CVA)-induced skeletal muscle damage, female rats were administered vehicle or CVA at 0.1, 0.5, and 1.0 mg/kg/day by gavage for two weeks and exercised or not on treadmills for 20 min/day. Clinical chemistry and plasma and tissue pharmacokinetics were evaluated; light and transmission electron microscopy (TEM) of Type I and Type II fiber-predominant skeletal muscles were performed. Serum levels of AST, ALT, CK, and plasma lactic acid were significantly elevated dose-dependently. CVA treatment decreased psoas and quadriceps weights. At 1 mg/kg all muscles except soleus demonstrated degeneration. Exercise-exacerbated severity of CVA-induced degeneration was evident in all muscles sampled except soleus and quadriceps. Early mitochondrial involvement in toxicity is suggested by the numerous membranous whorls and degenerate mitochondria observed in muscles at 0.5 mg/kg. No significant differences in CVA concentrations between either EDL and soleus or plasma and muscle were found.We found that CVA had no effect on cleaved caspase 3. In summary, we found that treadmill exercise exacerbated the incidence and severity of CVA-induced damage in Type II fiber-predominant muscles. Tissue exposure is likely not the key factor mediating CVA-induced skeletal muscle toxicity.

Xanya Sofra Weiss

Xanya Sofra Weiss

Abdominal Obesity, Impaired Nonesterified Fatty Acid Suppression, and Insulin-Mediated Glucose Disposal Are Early Metabolic Abnormalities in Families.

British Indian Asian men aged !40 years have a twofold to threefold increased risk of death from coronary heart
disease (CHD) compared with British whites. Epidemiological studies have suggested an association between glucose
intolerance and hyperinsulinemia with premature CHD in Indian Asians. We tested the association of insulin action with myocardial infarction (MI) by using the hyperinsulinemic-euglycemic clamp in 17 MI patients: 8 Punjabi Sikhs (PSMIs), 9 British whites (BWMIs), and 17 control subjects (9 PSCs and 8 BWCs). Metabolic factors associated with insulin resistance were investigated in 51 MI patients (24 PSMIs and 27 BWMIs) and 53 control subjects (28 PSCs and 25 BWCs). Familial aggregation of defective insulin action was examined by studying five pedigrees of Sikh survivors of MI. Sikh survivors of premature MI demonstrated impaired insulin-mediated glucose uptake (P!.001) by use of the clamp technique and nonesterified fatty acid (NEFA) suppression (P!.05) by using both clamp techniques and the oral glucose tolerance test, as compared with Sikh control subjects. White patients had impaired insulin-mediated glucose uptake but normal NEFA suppression. Metabolic factors usually associated with insulin resistance, including increased 2-hour post–oral glucose tolerance test triglycerides, smaller low density lipoprotein particle size, and increased plasminogen activator inhibitor-1, were present in white (all P!.05) but surprisingly absent in Sikh (all P".05) MI patients compared with respective ethnic control subjects. Fasting glucose and total cholesterol levels did not differ between patients and control subjects. Abdominal obesity, impaired NEFA suppression after oral glucose, and fasting hyperinsulinemia were present in Sikh MI patients and their nondiabetic first-degree relatives compared with Sikh control subjects. PS survivors of premature MI demonstrated impaired insulin-mediated glucose disposal and NEFA suppression compared with ethnic control subjects. BWMI patients showed abnormalities of carbohydrate, but not of NEFA, metabolism compared with white control subjects. Defects of insulin action manifested as abdominal obesity, impaired NEFA suppression, and fasting hyperinsulinemia are present in Sikh MI patients and their asymptomatic, nondiabetic, first-degree relatives.We suggest that these defects may be early metabolic markers that
predict risk of premature MI among PSs. (Arterioscler Thromb Vasc Biol. 1998;18:1021-1026.)

Xanya Sofra Weiss

Xanya Sofra Weiss

ELECTRICAL PROPERTIES OF INDIVIDUAL CELLS ISOLATED FROM ADULT RAT VENTRICULAR MYOCARDIUM. Xanya Sofra Weiss

The intricate anatomy of the myocardium, coupled with distinct morphology and function of specialized component cells, often presents acute problems in the inter- pretation of experimental data obtained from the whole organ. One experimental approach to minimize some of the difficulties encountered is the use of single cardiac cells isolated enzymically from adult mammalian myocardium and a technique has been developed in this laboratory whereby tissue dissociation is achieved by in vitro organ perfusion with crude bacterial collagenase (Gould & Powell, 1972;
Powell & Twist, 1976a). Myocytes isolated in this manner from adult rat heart have ntypical superficial and subcellular morphology (Gould & Powell, 1972; Powell, Steen, Twist & Woolf, 1978a), show respiratory coupling (Powell & Twist, 1975, 1976a) and have retained responsiveness to fi-adrenergic stimulation (Powell & Twist, 1976b; Powell, Terrar & Twist, 1978c). To date, however, there has been no systematic study of the electrical activity of these isolated cells, although such data are central to both the evaluation of the preparation as a useful experimental model and also to gain an insight into the electrophysiology of cardiac cells uncoupled physically from their usual syncitial state.
Experiments are presented to show that under suitable incubation conditions membrane potentials which are comparable to those reported for multicellular preparations can be recorded from isolated cells using intracellular micro-electrodes. It is also demonstrated that these cells can generate conventional action potentials, indicating that the relevant ionic mechanisms have been preserved during the iso- lation procedures. Some of these results have appeared previously in abstract form (Powell, Terrar & Twist, 1978b, c).

Xanya Sofra Weiss

Xanya Sofra Weiss

What Live Blood Cell Analysis Can Show You. Xanya Sofra Weiss

Normal Red Blood Cell (RBCs)

The circulatory system is the means by which oxygen, nutrients, antibodies, and hormones are transported to the cells to keep them alive and functioning. This is how your blood looks when you are experiencing optimum health. The Erythrocytes (cells) are round and separated and move through the capillaries very easily. The average size of
healthy RBC is 7.2 microns. The average number of red blood cells in the human body is 25 trillion. The average red blood cell lives 120 days. Your blood makes a complete circuit in your body every 30 seconds.

Protein Linkage

This condition is the first sign of cell stickiness and may progress into rouleau if not corrected. Protein linkage is a sign that excessive protein is being consumed or the protein is not being digested completely. As the cells start sticking together it becomes harder for your heart to push the blood through your veins and arteries.

Xanya Sofra Weiss

Xanya Sofra Weiss

Coupling of Calcium Homeostasis to Axonal Sodium in Axons of Mouse Optic Nerve. Xanya Sofra Weiss

YAKOV VERBNY, CHUAN-LI ZHANG, AND SHING YAN CHIU; 2002

Coupling of calcium homeostasis to axonal sodium in axons of mouse optic nerve. J Neurophysiol 88: 802–816, 2002; 10.1152/jn.00856.2001. Axonal populations in neonatal and mature optic nerves were selectively stained with calcium dyes for analysis of calcium homeostasis and its possible coupling to axonal Na. Repetitive nerve stimulation causes a rise in axonal [Ca2+]i the posttetanus recovery of which is impeded by increasing the number of action potentials in the tetanus. This effect is augmented in 4-aminopyridine (4-AP; 1 mM), which dramatically increases the calcium and presumably sodium load during the tetanus. Increasing axonal [Na]i with the Na-ionophore monensin (4–50 M) and ouabain (30 M) retards posttetanus calcium decline, suggesting that efficient calcium clearance depends on a low level of axonal [Na]i. the posttetanus recovery of which is impeded by increasing the number of action potentials in the tetanus. This effect is augmented in4-aminopyridine (4-AP; 1 mM), which dramatically increases the calcium and presumably sodium load during the tetanus. Increasing axonal [Na]i with the Na-ionophore monensin (4–50 M) and ouabain (30 M) retards posttetanus calcium decline, suggesting that efficient calcium clearance depends on a low level of axonal [Na]i. Posttetanus calcium clearance is not affected by K-mediated depolarization. To further examine coupling between axonal [Na]i and [Ca2+]i the resting axonal [Ca2+]i was monitored as axonal [Na+]i was elevated with ouabain, veratridine, and monensin. In all cases, elevation of axonal [Na+]i evokes a calcium influx into axons. This influx is unrelated to activation of calcium channels but is consistent with calcium influx via reversal of the Na/Ca exchanger expected as a consequence of axonal [Na+]i elevation. In conclusion, this study demonstrates that calcium homeostasis in the axons of the optic nerve is strongly coupled to axonal [Na+]i in a manner consistent with the Na/Ca exchanger playing a major role in extruding calcium following nerve activity.

Xanya Sofra Weiss

Xanya Sofra Weiss

CoQ10 and Statin Drugs. Xanya Sofra Weiss

Statin drugs have become very popular and are being widely prescribed in recent years to lower high blood cholesterol and thus reduce the risk for heart disease. These drugs block cholesterol production in the body by inhibiting the enzyme called HMG-CoA reductase in the early steps of its synthesis in the mevalonate pathway. This same biosynthetic pathway is also shared by CoQ10. Therefore, one unfortunate consequence of statin drugs is the unintentional inhibition of CoQ10 synthesis. Thus, in the long run, statin drugs could predispose the patients to heart disease by lowering their CoQ10 status, the very condition that these drugs are intended to prevent.Dr. Emile Bliznakov, an authority on CoQ10, recently published a scholarly review on the interaction between statin drugs and CoQ10 (Bliznakov and Wilkins, 1998). He wrote the best-selling book "The Miracle Nutrient Coenzyme Q10" several years ago and it is still being hailed as the best reference book on CoQ10 (Bliznakov, 1987).The reduction of CoQ10 levels might be associated with myopathy, a rare adverse effect associated with statin drugs. This metabolic myopathy is related to ubiquinone (CoQ10) deficiency in muscle cell mitochondria, disturbing normal cellular respiration and causing adverse effects such as rhabdomyolysis, exercise intolerance, and recurrent myoglobinuria.It is important to note that Coenzyme Q10 supplementation does not interfere with the very important cholesterol-lowering effect of statin drugs such as Lipitor and Zocor. Therefore, if you are taking a statin drug, (especially for an extended period of time), you may want to consider discussing CoQ10 supplementation with your health care professional. The bottom line is that the popular and widely prescribed cholesterol lowering drugs called "Statins" can block the synthesis of Coenzyme Q10 in the body which may lead to sub-optimal CoQ10 levels. Supplementation with Q-Gel CoQ-10 is a prudent approach when undergoing "statin" therapy.

Xanya Sofra Weiss

Xanya Sofra Weiss

AIDS-Related Dementia and Calcium Homeostasis. Xanya Sofra Weiss

STUART A. LIFTON

Our laboratory has a long-standing interest in the relationship of neuronal viability and outgrowth to intracellular Ca2+ levels (reviewed in ref. 1). Glutamate, or a related excitatory amino acid (EAA), is the major excitatory neurotransmitter that controls the level of intracellular neuronal Ca"Our laboratory has a long-standing interest in the relationship of neuronal viability and outgrowth to intracellular Ca2+ levels (reviewed in ref. 1). Glutamate, or a related excitatory amino acid (EAA), is the major excitatory neurotransmitter that controls the level of intracellular neuronal Ca2+([Ca2+]i). Escalating concentrations of glutamate have been measured in vivo following focal stroke and head injury (reviewed in refs. 2 and 3). As a result, there is a rapid rise in [Ca2+]i CaZtli may not account by itself for the ensuing neuronal injury, several laboratories have now reported that prevention of the increase in [Ca2+]i Jlie ads to amelioration of anticipated neuronal cell death (reviewed in refs. 2 and 3). Excessive intracellular Ca2+ is thought to contribute to the triggering of a series of potentially neurotoxic events leading to cellular necrosis orperhaps apoptosis. Many mechanisms are involved in intracellular calcium homeostasis, and this subject is beyond the scope of this article (but see the review in ref. 4).Here we will consider only two modes of Caz+ entry into neurons during these pathological processes. These two routes of entry of Ca2+o ccur via ion channels that are permeable to Ca2+ and can be summarized as follows: (1) Glutamate or related EAAs trigger voltage-dependent calcium channels by depolarizing the cell membrane; a major voltage-dependent calcium channel subtype that is chronically activated by prolonged depolarizations is the L-type calcium channel (reviewed in ref. 5). (2) Glutamate or related EAAs activate ligand-gated ion channels directly; a predominant glutamate receptor-operated channel that is permeable to Ca2+ under these conditions is the Nmethyl-D-aspartate (NMDA) subtype, but other non-NMDA types may also contribute (reviewed in refs. 2 and 6). We have shown that activation of these channel types can control neuronal plasticity during normal development, but our laboratory and many others have shown that in excessive amounts this stimulation can lead to neuronal death, for example, after a stroke (reviewed in ref. 1). Similar mechanisms may obtain in various neurodegenerative conditions. In fact, although not involved in the primary pathophysiology of a neurologic disorder, this mechanism may represent a final common pathway of neuronal injury. Most importantly, this pathway makes the disease process amenable to pharmacotherapy. This line of reasoning led us to think that this mechanism might be involved in acquired immunodeficiency syndrome (AIDS)-related neuronal injury.

Xanya Sofra Weiss

Xanya Sofra Weiss

Inhibitory effect of green coffee bean extract on fat accumulation and body weight gain in mice. Xanya Sofra Weiss

Hiroshi Shimoda, Emi Seki, and Michio Aitani; 2006

An epidemiological study conducted in Italy indicated that coffee has the greatest antioxidant capacity among the commonly consumed beverages. Green coffee bean is rich in chlorogenic acid and its related compounds. The effect of green coffee bean extract (GCBE) on fat accumulation and body weight in mice was assessed with the objective of investigating the effect of GCBE on mild obesity.
Methods: Male ddy mice were fed a standard diet containing GCBE and its principal constituents, namely, caffeine and chlorogenic acid, for 14 days. Further, hepatic triglyceride (TG) level was also investigated after consecutive administration (13 days) of GCBE and its constituents. To examine the effect of GCBE and its constituents on fat absorption, serum TG changes were evaluated in olive oil-loaded mice. In addition, to investigate the effect on hepatic TG metabolism, carnitine palmitoyltransferase (CPT) activity in mice was evaluated after consecutive ingestion (6 days) of GCBE and its constituents (caffeine, chlorogenic acid, neochlorogenic acid and feruloylquinic acid mixture) Results: It was found that 0.5% and 1% GCBE reduced visceral fat content and body weight. Caffeine and chlorogenic acid showed a tendency to reduce visceral fat and body weight. Oral administration of GCBE (100 and 200 mg/kg· day) for 13 days showed a tendency to reduce hepatic TG in mice. In the same model, chlorogenic acid (60 mg/kg· day) reduced hepatic TG level. In mice loaded with olive oil (5 mL/kg), GCBE (200 and 400 mg/kg) and caffeine (20 and 40 mg/kg) reduced serum TG level. GCBE (1%), neochlorogenic acid (0.028% and 0.055%) and feruloylquinic acid mixture (0.081%) significantly enhanced hepatic CPT activity in mice. However, neither caffeine nor chlorogenic acid alone was found to enhance CPT activity.
Conclusion: These results suggest that GCBE is possibly effective against weight gain and fat accumulation by inhibition of fat absorption and activation of fat metabolism in the liver. Caffeine was found to be a suppressor of fat absorption, while chlorogenic acid was found to be partially involved in the suppressive effect of GCBE that resulted in the reduction of hepatic TG level. Phenolic compounds such as neochlorogenic acid and feruloylquinic acid mixture, except chlorogenic acid, can enhance hepatic CPT activity.

Xanya Sofra Weiss

Xanya Sofra Weiss

About Arasys Perfector. Xanya Sofra Weiss

Arasys Perfector, LLC is the Global distributor of the UK-based Anti-Aging Technology of Ion Magnum, Arasys, and Perfector. These three technologies share one vision: Working with the body’s own resources to prevent aging, enhance biological functions, well-being and longevity. Arasys Perfector, LLC has been conducting its own research and development since 2000. Most of our profit goes into an on-going effort of collaboration with anti-aging experts and doctors to improve the amazing capabilities of our micro/nano/pico current devices. Arasys Perfector, LLC began providing products, training, and solutions to businesses in the USA since 2004 and has begun partnership distributions in other parts of the world such as Asia, Canada, and Dubai starting in 2008.

We’ve developed this weblog to keep our current and prospective customers involved with the on-going research surrounding the Arasys, Perfector, and Ion Magnum devices.

Xanya Sofra Weiss

Xanya Sofra Weiss

Aging Progressively Impairs Endothelium-Dependent Vasodilation in Forearm Resistance Vessels of Humans. Xanya Sofra Weiss

Studies in experimental models suggest that endothelium-derived nitric oxide is reduced with aging, and this circumstance may be relevant to atherogenesis. The aim of this study was to determine whether increasing age resulted in altered endothelium-dependent vasodilation in the forearm resistance vessels of healthy humans. Forearm blood flow was measured in 119 healthy subjects, aged 19 to 69 years, by venous occlusion plethysmography. Brachial artery infusions of methacholine chloride (0.03 to 10.0 µg/min) were used to assess endothelium-dependent vasodilation and of sodium nitroprusside (0.03 to 10.0 µg/min) to assess endothelium-independent vasodilation. The slope of the dose–blood flow response relation was calculated in each subject for each drug. Univariate and multiple stepwise regression analyses were used to relate vascular reactivity to selected variables, including age, lipids, and blood pressure. Endothelium-dependent vasodilation was progressively impaired with increasing age, assessed as a reduction in slope from 2.25±0.16 to 0.34±0.11 (mL/100 mL tissue per minute)/(µg/min) (P<.001). The decline in endothelium-dependent vasodilation was already evident by the fourth decade (age 30 to 39 years). Endothelium-independent vasodilation did not change with age. Age, total cholesterol, and low-density lipoprotein cholesterol were univariate predictors of endothelium-dependent vasodilation. Age remained the most significant predictor of endothelium-dependent vasodilator responses by multiple stepwise regression analysis. From these observations, it can be concluded that endothelium-dependent vasodilation declines steadily with increasing age in healthy human subjects. Age is a strong univariate and multivariate predictor of endothelium-dependent vasodilation. This finding may be a marker for more widespread endothelial dysfunction.

Xanya Sofra Weiss

Xanya Sofra Weiss

PILOT STUDY OF IMPEDANCE-CONTROLLED MICROCURRENT THERAPY FOR MANAGING RADIATION-INDUCED FIBROSIS IN HEAD-AND-NECK CANCER PATIENTS. Xanya Sofra Weiss

As aggressive therapy with combination surgery, chemotherapy, and radiotherapy (RT) increases tumor control in head-and-neck neoplasms, posttreatment quality-of-life issues remain problematic (1). One area of concern is progressive fibrosis of soft tissue in the head, neck, and supraclavicular area. For many patients, palpation of the treated areas reveals hard, unyielding tissue that limits range of motion and/or leads to pain associated with movement. The concept of investigating microcurrent therapy to treat radiation-induced fibrosis arose from the observation of a salivary gland patient who was receiving microcurrent therapy for the surgical scar at a family physician’s office while receiving neutron therapy at Fermilab. The patient experienced significantly milder erythema and mucositis than would historically be expected for radical RT in the neck area. This serendipitous observation led to a hypothesis that microcurrent therapy could be beneficial in managing the effects of RT.

Xanya Sofra Weiss

Xanya Sofra Weiss

Effects of weight loss on liver and erythrocyte polyunsaturated fatty acid pattern and oxidative stress status in obese patients with non-alcoholic.

Our aim was to study the influence of weight loss on the fatty acid (FA) composition of liver and erythrocyte phospholipids and oxidative stress status in obese, non-alcoholic, fatty liver disease (NAFLD) patients. Seven obese NAFLD patients who underwent subtotal gastrectomy with a gastro-jejunal anastomosis in roux and Y were studied immediately and 3 months after surgery. Seven non-obese patients who underwent anti-reflux surgery constituted the control group. Serum F2-isoprostane levels were measured by GS/NICI-MS/MS and FA composition was determined by GC. At the time of surgery, controls and obese patients exhibited a hepatic polyunsaturated fatty acid (PUFA) pattern that correlated with that of erythrocytes. Three months after surgery, NAFLD patients lost 21% of initial body weight; serum F2-isoprostane levels decreased by 76%; total PUFA, long-chain PUFA (LCPUFA), n-3 PUFA, and n-3 LCPUFA increased by 22, 29, 81, and 93%, respectively; n-6/n-3 LCPUFA ratio decreased by 51%; docosahexaenoic acid/docosapentaenoic acid ratio increased by 19-fold; and the n-3 product/precursor ratio (20: 5 + 22: 5 + 22: 6)/18: 3 increased by 164% (p<0.05). It is concluded that weight loss improves the n-3 LCPUFA status of obese patients in association with significant amelioration in the biomarkers of oxidative stress, membrane FA insaturation, and n-3 LCPUFA biosynthesis capacity, thus representing a central therapeutic issue in the improvement of obesity-related metabolic alterations involved in the mechanism of hepatic steatosis.

Xanya Sofra Weiss

Xanya Sofra Weiss

A single mutation in the castor 9-18:0-desaturase changes reaction partitioning from desaturation to oxidase chemistry. Xanya Sofra Weiss

A single mutation in the castor 9-18:0-desaturase changes reaction partitioning from desaturation to oxidase chemistry Jodie E. Guy*, Isabel A. Abreu†, Martin Moche‡, Ylva Lindqvist*, Edward Whittle†, and John Shanklin†§ *Department of Medical Biochemistry and Biophysics, Division of Molecular Structural Biology, Karolinska Institutet, Tomtebodava¨gen 6, S-171 77 Stockholm, Sweden; †Department of Biology, Brookhaven National Laboratory, Upton, NY 11973; and ‡Department of Medical Biochemistry and Biophysics and Structural Genomics Consortium, Karolinska Institutet, S-171 77 Stockholm, Sweden Edited by Christopher R. Somerville, Carnegie Institution of Washington, Stanford, CA, and approved September 28, 2006 (received for review August 17, 2006) Sequence analysis of the diiron cluster-containing soluble desaturases suggests they are unrelated to other diiron enzymes; however, structural alignment of the core four-helix bundle of desaturases to other diiron enzymes reveals a conserved iron binding motif with similar spacing in all enzymes of this structural class, implying a common evolutionary ancestry. Detailed structural comparison of the castor desaturase with that of a peroxidase, rubrerythrin, shows remarkable conservation of both identity and geometry of residues surrounding the diiron center, with the exception of residue 199. Position 199 is occupied by a threonine in the castor desaturase, but the equivalent position in rubrerythrin contains a glutamic acid. We previously hypothesized that a carboxylate in this location facilitates oxidase chemistry in rubrerythrin by the close apposition of a residue capable of facilitating proton transfer to the activated oxygen (in a hydrophobic cavity adjacent to the diiron center based on the crystal structure of the oxygen-binding mimic azide). Here we report that desaturase mutant T199D binds substrate but its desaturase activity decreases by 2 103-fold. However, it shows a &gt;31-fold increase in peroxide-dependent oxidase activity with respect toWT desaturase, as monitored by single-turnover stopped-flow spectrometry. A 2.65-Å crystal structure of T199D reveals active-site geometry remarkably similar to that of rubrerythrin, consistent with its enhanced function as an oxidase enzyme. That a single amino acid substitution can switch reactivity from desaturation to oxidation provides experimental support for the hypothesis that the desaturase evolved from an ancestral oxidase enzyme. binuclear diiron enzyme Nonheme diiron-containing four-helix-bundle proteins possess the ability to functionalize unactivated C-H groups and mediate a diversity of chemical reactions including oxidation, hydroxylation, desaturation, and epoxidation (1, 2). A wealth of mechanistic information is available from various diironcontaining proteins including methane monooxygenases, 9 desaturases, ribonucleotide reductases, rubrerythrins, alternate oxidases, ferritins, and bacterioferritins (1–3). The diiron-containing proteins are highly divergent in their amino acid sequences, with identities typically falling below that necessary for conventional phylogenetic analysis. However, when the analysis is restricted to the four helices that coordinate the diiron active site, the amino acid identity rises to 16–31% (4). A shared diiron-binding motif within the conserved four-helix bundle is involved in oxygen chemistry. The reactions have been described as occurring in two phases, an oxygen activation phase followed by reaction phases (1). Oxygen activation likely placed strong evolutionary constraints on the organization of the diiron center, whereas the reaction phases exhibit great diversity of functional outcome. In addition to their individual catalytic reactions, rubrerythrin, methane monooxygenase, ribonucleotide reductase, and the 9 desaturase have also been shown to reduce dioxygen to water (4–6). Based on these similarities, Gomes et al. (4) proposed that the four-helix bundle diiron proteins arose from a common ancestor that bound activated oxygen species and reduced them to water. This hypothetical oxidase enzyme is thought to have appeared at the transition from anaerobic to aerobic environment, 2.5 billion years ago. We previously performed a structural comparison of the active site of the 9 desaturase with that of rubrerythrin, an NAD(P)H peroxidase, which revealed remarkable similarity of the diiron ligands (7). Based on this structural analysis we proposed that residue 199, which occupies a location adjacent to the diiron site and abuts the hydrophobic substrate binding cavity, plays a key role in determining the chemical outcome of the enzyme (7). In the desaturase it is occupied by threonine, and in the rubrerythrin it is occupied by a glutamic acid. In this work we report that the T199D mutant of the 9 desaturase shows greatly reduced desaturation activity but increases its oxidase activity by 31-fold with respect to theWT desaturase. A crystal structure of the T199D mutant is presented that shows very close active-site similarity to rubrerythrin, consistent with its change in functionality. Results and Discussion Structural alignment of the reduced azide complexes of 9 desaturase and rubrerythrin (8) revealed similarities with respect to the position and identity of iron binding ligands and the position of the azide adduct (7) (Fig. 1). The single major difference in the active site is the identity of the residue corresponding to threonine-199 in the desaturase, which is a glutamic acid in rubrerythrin. The side chain of the residue occupying this position faces the bound azide that mimics the binding site of molecular oxygen. Thus, the desaturase contains threonine, a poor proton donor, whereas rubrerythrin contains a glutamic acid, which facilitates proton transfer. We previously hypothesized that the presence or absence of a proton donor in this position might influence the partitioning of chemical reactivity of the diiron site between desaturation and oxidase chemistry (7). Thus, we engineered mutations at position 199 into the desaturase to replace threonine with either glutamic or aspartic acid and compared the desaturase and oxidase activity of these mutants to those of WT 9 desaturase.

Xanya Sofra Weiss

Xanya Sofra Weiss

Systems Biology, Toxins, Obesity, and Functional Medicine. Xanya Sofra Weiss

13th International Symposium of The Institute for Functional Medicine S 134 Managing Biotransformation: The Metabolic, Genomic, and Detoxification Balance Points Hyman Mark Hyman, MD, is editor in chief of Alternative Therapies in Health and Medicine, medical editor of Alternative Medicine, the Art and Science of Healthy Living, and the author of several books. He is also on the Board of Advisors and faculty of Food as Medicine, Center for Mind Body Medicine, Georgetown University School of Medicine, and on the Board of Directors and faculty of the Institute for Functional Medicine, as well as collaborating with the Harvard Medical School’s Division for Research and Education in Complementary and Integrative Medicine. Obesity is not a single clinical disorder. Obesity is a complex chronic illness resulting from the interplay among genetics, environment, and lifestyle. Emerging scientific concepts provide a new basis for understanding the multiple causes of obesity as well as the underlying mechanisms involved in weight dysregulation. While most obesity can be effectively treated for compliant patients, using a focused lifestyle intervention based on a whole-foods, low-glycemicload, phytonutrient-rich diet combined with exercise and stress management, there are patients who do not respond predictably to normally successful interventions. A novel hypothesis linking environmental and internal toxins to disruptions of key mechanisms involved in weight regulation may explain treatment resistance in obesity. The key biological systems involved in obesity (and all diseases) that are altered by toxins are the neuro-endocrine-immune system, and mitochondrial energetics and redox status. Obesity provides an illustrative example of new navigational tools for diagnosis and therapy of chronic illness based on a paradigm that focuses not on disease or symptoms, but on cause and mechanism. This new framework and methodological approach can be applied to any chronic disease and provides an opportunity to integrate fragmentary scientific discoveries into a cohesive whole that creates a new clinical roadmap. This paper will explore a novel hypothesis that links obesity and toxins; we will discuss how one particular disease and the effect of one underlying cause can create a clinically relevant, holographic view of physiology. Alterations in thyroid metabolism and receptor function, central appetite dysregulation, inflammation’s influence on insulin and leptin resistance, impaired mitochondrial oxidative metabolism, and oxidative-stress-mediated effects via nuclear factor kappa B (NFκB) are all mechanisms by which toxins create alterations in metabolism and finely-tuned weight regulatory mechanisms. These systems are not discrete entities but systems in the true sense of the word – interlocking, interactive, dynamic, overlapping networks of biochemical and physiological informational spheres of functional relationships. Multiple patterns of genetic, physiological, and biochemical dysfunction are linked to obesity, including genetic polymorphisms, inflammation, mitochondrial dysfunction, oxidative stress, neuro-endocrine-immune dysfunction, especially autonomic disturbances involving the hypothalamic-pituitary-adrenal axis, nutritional deficiencies or excesses, and toxins. The nature, causes, and remediation of obesity can be seen through the prism of any one of these patterns. The focus here will be on how toxins mediate their influence through all these mechanisms.

Xanya Sofra Weiss

Xanya Sofra Weiss

Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase. Xanya Sofra Weiss

Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase Qiming Suna, Weiliang Fana, Keling Chena, Xiaojun Dingb, She Chenb, and Qing Zhonga,1 aDivision of Biochemistry and Molecular Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720; and bNational Institute of Biological Sciences, Beijing 102206, China Communicated by Xiaodong Wang, University of Texas Southwestern Medical Center, Dallas, TX, October 17, 2008 (received for review October 10, 2008) Autophagy mediates the cellular response to nutrient deprivation, protein aggregation, and pathogen invasion in human. Dysfunction of autophagy has been implicated in multiple human diseases including cancer. The identification of novel autophagy factors in mammalian cells will provide critical mechanistic insights into how this complicated cellular pathway responds to a broad range of challenges. Here, we report the cloning of an autophagy-specific protein that we called Barkor (Beclin 1-associated autophagy-related key regulator) through direct interaction with Beclin 1 in the human phosphatidylinositol 3-kinase class III complex. Barkor shares 18% sequence identity and 32% sequence similarity with yeast Atg14. Elimination of Barkor expression by RNA interference compromises starvation- and rapamycin-induced LC3 lipidation and autophagosome formation. Overexpression of Barkor leads to autophagy activation and increased number and enlarged volume of autophagosomes. Tellingly, Barkor is also required for suppression of the autophagy-mediated intracellular survival of Salmonella typhimurium in mammalian cells. Mechanistically, Barkor competes with UV radiation resistance associated gene product (UVRAG) for interaction with Beclin 1, and the complex formation of Barkor and Beclin1 is required for their localizations to autophagosomes. Therefore, we define a regulatory signaling pathway mediated by Barkor that positively controls autophagy through Beclin 1 and represents a potential target for drug development in the treatment of human diseases implicated in autophagic dysfunction. Atg14 autophagosome LC3 Salmonella UVRAG One of the central regulators of autophagy in mammalian cells is Beclin 1 (1–3). Beclin 1 is a component of the class III phosphatidylinositol 3-kinase (PI3KC3) complex, which also contains a PI3K catalytic subunit and a regulatory subunit (p150) (4). Beclin 1 was identified as a haploid insufficient tumor suppressor gene (3). It is monoallelically deleted in ovarian, breast, and prostate cancers. Heterozygous Beclin 1 / mice have reduced autophagy activity and increased incidence of spontaneous tumors (5, 6). Allelic loss of Beclin 1 leads to genome instability upon metabolic stress (7, 8). All of this evidence illustrates a role for Beclin 1 and autophagy in cancer development. Notably, Beclin 1 and PI3KC3 have pleiotropic functions in multiple cellular processes. PI3KC3 is not only required for autophagy, but also has broad functions in endocytic protein sorting (9). Functional equivalents of Beclin 1/PI3KC3/p150 in yeast, Vps30/ Atg6-Vps15-Vps34, are known to play a critical role in autophagy and in vacuolar protein sorting (VPS) (1, 10). The specificity of PI3KC3 in yeast is determined by different complex compositions. Two regulatory proteins, Atg14 and Vps38, direct the core PI3K complex to either the phagophore assembly site (PAS) for autophagy or the endosome for VPS (10, 11), respectively, to execute their functions in autophagy or VPS. Atg14 is required for mediating the localization of the core PI3KC3 complex to PAS and is also important in recruiting downstream Atg proteins such as Atg2, Atg8, Atg16, and the Atg12-Atg5 conjugate to the PAS for membrane elongation and vesicle completion (12, 13). In contrast, Vps38 is responsible for the endosomal localization of the PI3K complex (11). Surprisingly, such regulatory mechanisms directing PI3KC3 specificity have not been identified in mammals. How the function of Beclin 1 is specifically directed toward autophagosomes in mammalian cells has remained elusive. We speculate that there are autophagy-specific factors mediating Beclin 1 activity in autophagy. We used a biochemical approach to purify and proteomic methods to characterize the Beclin 1 complex. Here, we report the identification of a Beclin 1- associated protein that promotes autophagy specifically through the interaction with Beclin 1. Results Identification of Barkor as a Beclin 1-Interacting Protein. To search for Beclin 1 regulatory proteins, we generated a cell line from human osteosarcoma U2OS cells that is stably transfected with ZZ-Beclin 1-FLAG under the control of doxycycline [supporting information (SI) Fig. S1A]. The expression of Beclin 1 was adjusted by the titration of doxycycline, and a dose (20 ng/mL) that induces expression of tagged Beclin 1 close to the endogenous level was selected (Fig. S1B). The tagged Beclin 1 was purified from cell extracts by sequential affinity chromatography steps, and the final FLAGpeptide eluate was subjected to 4–12% gradient SDS/PAGE and visualized by silver staining (Fig. 1A). The indicated bands were excised and analyzed by mass spectrometry. In addition to the known components of the Beclin 1 complex, namely the PI3K catalytic subunit, p150 regulatory subunit, and UVRAG, we also identified a 68-kDa protein by mass spectrometry, KIAA0831 (Fig. 1A), which we called Barkor (Beclin 1-associated autophagy related key regulator). We were able to purify the same complex from human embryonic kidney 293T cells expressing tagged Beclin 1, indicating that the formation of this complex is not cell type-specific (Fig. 1B). Bioinformatic analysis revealed that Barkor contains an N-terminal zinc finger motif and a central coiled-coil domain (CCD) (Fig. S2) and a domain organization similar to Atg14 in yeast. Barkor also shares 18% sequence identity and 32% sequence similarity with yeast Atg14 (Fig. S3). The identities of these interacting proteins were further confirmed by immunoblotting analysis (Fig. S4). Although another Beclin 1-interacting protein, Bcl-2 (14), could not be visualized by silver staining, its presence in the final eluate was validated by immunoblotting (Fig. S4). The interaction of Barkor and Beclin 1 was further confirmed by the Author contributions: Q.Z. designed research; Q.S., W.F., and K.C. performed research; X.D. and S.C. contributed new reagents/analytic tools; Q.S., W.F., and Q.Z. analyzed data; and Q.S. and Q.Z. wrote the paper. The authors declare no conflict of interest. Freely available online through the PNAS open access option. 1Towhomcorrespondence should be addressed at: Department of Molecular and Cell Biology, University of California, 316 Barker Hall, Berkeley, CA.

Xanya Sofra Weiss

Xanya Sofra Weiss

The prosurvival role of autophagy in Resveratrol-induced cytotoxicity in human U251 glioma cells. Xanya Sofra Weiss

Previous study reported that resveratrol has anti-tumor activity. In this study, we investigated the involvement of autophagy in the resveratrol-induced apoptotic death of human U251 glioma cells. METHODS: The growth inhibition of U251 cells induced by resveratrol was assessed with methyl thiazolyl tetrazolium (MTT). The activation of autophagy and proapoptotic effect were characterized by monodansylcadaverine labeling and Hoechst stain, respectively. Mitochondrialtransmembrane potential (DeltaPsim) was measured as a function of drug treatment using 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). The role of autophagy and apoptosis in the resveratrol-induced death of U251 cells was assessed using autophagic and caspase inhibitors. Immunofluorescence, flow cytometry, and Western blot analysis were used to study the apoptotic and autophagic mechanisms. RESULTS: Methyl thiazolyl tetrazolium (MTT) assays indicated that resveratrol decreased the viability of U251 cells in a dose- and time-dependent manner. Flow cytometry analysis indicated that resveratrol increased cell population at sub-G1 phase, an index of apoptosis. Furthermore, resveratrol-induced cell death was associated with a collapse of the mitochondrial membrane potential. The pan-caspase inhibitor Z-VAD-fmk suppressed resveratrol-induced U251 cell death. Resveratrol stimulated autophagy was evidenced by punctuate monodansylcadaverine(MDC) staining and microtubule-associated protein light chain 3 (LC3) immunoreactivty. Resveratrol also increased protein levels of beclin 1 and membrane form LC3 (LC3-II). Autophagy inhibitors 3-methylademine (3-MA) and bafilomycin A1 sensitized the cytotoxicity of resveratrol. CONCLUSION: Together, these findings indicate that resveratrol induces autophagy in human U251 glioma cells and autophagy suppressed resveratrol-induced apoptosis. This study thus suggests that autophagy inhibitors can increase the cytotoxicity of resveratrol to glioma cells. BMC Cancer. 2009 Jun 30;9:215

Xanya Sofra Weiss

Xanya Sofra Weiss

Circulating free T3 in pregnancy, liver diseases, diabetes mellitus and thyroid diseases. Xanya Sofra Weiss

Nippon Naibunpi Gakkai Zasshi ; 1984 Measurement of serum concentrations of free triiodothyronine (FT3) is considered to be an accurate index of thyroid function in the patient. In this study, we measured serum concentrations of FT3, free thyroxine (FT4) and reverse triiodothyronine (rT3) by radioimmunoassay in blood samples taken from the navel cord of 20 newborns as well as 20 nonpregnant women, 20 pregnant women, 10 patients with liver diseases, 25 patients with diabetes mellitus, 65 patients with hyperthyroidism, 30 patients with primary hypothyroidism and 29 normal subjects. In pregnant women, serum FT3 and FT4 levels gradually decreased as the pregnancy progressed. In cord blood, FT3 levels were less than a quarter of the values found during the first trimester of pregnancy or that of non-pregnant women, whereas serum rT3 levels were drastically increased. In chronic hepatitis, liver cirrhosis and diabetes mellitus, serum FT3 and FT4 levels were significantly lower than that in the controls. In thyroid diseases, serum FT3 levels varied parallel to other thyroid hormone levels. In primary hypothyroidism, however, serum FT3 levels were still lower than these in the controls after treatment with 1-thyroxine, whereas other thyroid hormone levels and TSH levels returned to control levels.

Xanya Sofra Weiss

Xanya Sofra Weiss

Free Radicals in Biology. Xanya Sofra Weiss

Free radicals play an important role in a number of biological processes, some of which are necessary for life, such as the intracellular killing of bacteria by neutrophil granulocytes. Free radicals have also been implicated in certain cell signalling processes ^[5]. This is dubbed redox signaling. The two most important oxygen-centered free radicals are superoxide and hydroxyl radical. They are derived from molecular oxygen under reducing conditions. However, because of their reactivity, these same free radicals can participate in unwanted side reactions resulting in cell damage. Many forms of cancer are thought to be the result of reactions between free radicals and DNA, resulting in mutations that can adversely affect the cell cycle and potentially lead to malignancy. Some of the symptoms of aging such as atherosclerosis are also attributed to free-radical induced oxidation of many of the chemicals making up the body. In addition free radicals contribute to alcohol-induced liver damage, perhaps more than alcohol itself. Radicals in cigarette smoke have been implicated in inactivation of alpha 1-antitrypsin in the lung. This process promotes the development of emphysema. Free radicals may also be involved in Parkinson's disease, senile and drug-induced deafness, schizophrenia, and Alzheimer's. The classic free-radical syndrome, the iron-storage disease hemochromatosis, is typically associated with a constellation of free-radical-related symptoms including movement disorder, psychosis, skin pigmentary melanin abnormalities, deafness, arthritis, and diabetes mellitus. The free radical theory of aging proposes that free radicals underlie the aging process itself, whereas the process of mitohormesis suggests that repeated exposure to free radicals may extend life span. Because free radicals are necessary for life, the body has a number of mechanisms to minimize free radical induced damage and to repair damage which does occur, such as the enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. In addition, antioxidants play a key role in these defense mechanisms. These are often the three vitamins, vitamin A, vitamin C and vitamin E and polyphenol antioxidants. Further, there is good evidence bilirubin and uric acid can act as antioxidants to help neutralize certain free radicals. Bilirubin comes from the breakdown of red blood cells' contents, while uric acid is a breakdown product of purines. Too much bilirubin, though, can lead to jaundice, which could eventually damage the central nervous system, while too much uric acid causes gout ^[6].

Xanya Sofra Weiss

Xanya Sofra Weiss

Glutathione: Health Info. Xanya Sofra Weiss

Nevertheless, people have tried glutathione for the treatment of a whole host of conditions, including cancer, high blood pressure, Parkinson's disease, Alzheimer's disease, cataracts, and male infertility.

The best studies have been conducted in cancer. One study involved women with ovarian cancer who were being treated with chemotherapy. Some of the women were also treated with intravenous glutathione. Those given the glutathione not only had fewer side effects from the chemotherapy but also had better overall survival rates.

Myriam Abalain of Montreal, Canada, is one of the many people who have taken Bounous's Immunocal to combat cancer. In 1996, at age 33, a routine PAP smear revealed she had precancerous cells on her cervix, which is one step away from having cervical cancer. The three specialists she visited all told her that a hysterectomy was her only option, but she hesitated to have such major, life-altering surgery.

Instead, she waited. For more than two years, her condition remained stable. Then a friend suggested she try Immunocal. After eight months of taking the supplement, her physician could no longer detect any precancerous cells. Does this mean Immunocal cured her? It's hard to say based on just one case like hers. It is possible her body went into remission naturally.

Even Bounous acknowledges there's no real proof his product cured her cancer, but he's working on conducting good clinical research, comparing individuals with cancer taking glutathione to those who are not.

What Are the Risks?

Overall, taking glutathione or its precursors in reasonable amounts appears to be quite safe, although it should be avoided in people with milk protein allergies and in those who have received an organ transplant. There is also some concern, however, about the safety of taking glutathione for the one condition for which there is the greatest evidence of its usefulness: cancer.

"People don't get concerned about these health-promoting [supplements] until they're in their 50s and 60s," says Emory's Dean Jones. At that point, they may already have the initial precancerous [cells]. Therefore, the supplements, just like they promote health in normal tissues, might promote health in the precancerous tissue."

Appleton recognizes this possibility but says "there's no evidence that supplementing with glutathione, even intravenously, is in any way going to make any cancer worse. In fact, the evidence we have suggests the opposite. It suggests that glutathione and other antioxidants, far from interfering with the activity of chemotherapy, appear to reduce side effects without decreasing efficacy and may, in fact, improve the efficacy of the chemotherapy in fighting cancer."

Bounous says his research has demonstrated that taking Immunocal actually lowers glutathione in cancer cells while increasing it in normal cells. As a result, the cancer cells are more vulnerable to chemotherapy, and the normal cells are protected.

The upshot? The experts disagree on who should take glutathione or its precursors. Bounous says everyone should take it in order to optimize overall health. Appleton would reserve it for people with cancer. Jones says it might only prove beneficial for those who eat poorly and are thus unlikely to be getting much glutathione or its precursors in their diet.

They all acknowledge that people with severe diseases known to be associated with low glutathione levels, such as AIDS, may well benefit from the supplement, although there is no proof to this effect.

For her part, Myriam Abalain is still taking Immunocal and feeling fine. "I'm doing pretty good now," she says. "I'm in better shape than ever!"

Xanya Sofra Weiss

Xanya Sofra Weiss

Resveratrol and Cancer. Xanya Sofra Weiss

Resveratrol (3,5,4-trihydroxystilbene), a natural phytoalexin present in grapes, nuts, and red wine, has antineoplastic activities. Several molecular mechanisms have been described to underlie its effects on cells in vitro and in vivo. In the present study, the response of ovarian cancer cells to resveratrol is explored. Resveratrol inhibited growth and induced death in a panel of five human ovarian carcinoma cell lines. The response was associated with mitochondrial release of cytochrome c, formation of the apoptosome complex, and caspase activation. Surprisingly, even with these molecular features of apoptosis, analysis of resveratrol-treated cells by light and electron microscopy revealed morphology and ultrastructural changes indicative of autophagocytic, rather than apoptotic, death. This suggests that resveratrol can induce cell death through two distinct pathways. Consistent with resveratrol’s ability to kill cells via nonapoptotic processes, cells transfected to express high levels of the antiapoptotic proteins Bcl-xL and Bcl-2 are equally sensitive as control cells to resveratrol. Together, these findings show that resveratrol induces cell death in ovarian cancer cells through a mechanism distinct from apoptosis, therefore suggesting that it may provide leverage to treat ovarian cancer that is chemoresistant on the basis of ineffective apoptosis.

Xanya Sofra Weiss

Xanya Sofra Weiss

Direct-Current Bactericidal Effect on Intact Skin. Xanya Sofra Weiss

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, JUlY 1980, P. 137-141 Vol. 18, No. 1 0066-4804/80/07-0137/05$02.00/0 Direct-Current Bactericidal Effect on Intact Skin L. BOLTON,`* B. FOLENO,' B. MEANS, and S. PETRUCELLI2 Johnson & Johnson Products, Inc., New Brunswick, New Jersey 08903,1 and ESB, Inc., Division of The International Nickel Company of Canada, Limited, Madison, Wisconsin 537142 Positive carbon-containing electrodes conveying 5 or more ,uA of constant direct current per cm2 showed bactericidal activity on intact back skin of 13 human subjects. This effect increased with the duration of stimulation up to a total surface bacterial kill at 20 h. When total current and current density were varied independently on 16 sites on the backs of eight subjects, the effect was dependent on current density, not on total current. Electrodes driven by similar voltages but which removed the electrochemical reaction from inoculated sites on the backs of three subjects failed to reduce the numbers of colony-forming units as compared with those sampled from control sites. This showed the bactericidal effect to be electrochemical in origin, probably mediated by local acidity generated at the surface of the positive carbon-containing electrodes. With an adhesive tape stripping technique on three sites on each of six subjects, it was determined that the effect extended into the epidermis of the human back. No effect was observed beneath negative or control electrodes under the same conditions. Direct-current stimulation of a biological subject may produce effects resulting from ion flow within the subject or from electrochemical events at the tissue-electrode interface or both. Metallic ions released electrochemically at positive electrodes have been reported to exhibit local antimicrobial activity (1, 2, 4-7). However, little is known about the effects of direct current delivered through nonmetallic carbon electrodes. The present study investigated the effects of constant direct-current stimulation through carbon- filled electrodes on microorganisms in intact human skin. MATERIALS AND METHODS Electrical apparatus. Two types of stimulating electrodes were used. In the first type of electrode, Condulon film (Bemis Co., Inc., Minneapolis, Minn.), a polyvinyl chloride film containing carbon, served as the electrode surface on a 6-V flat-pack dry cell. This battery-electrode complex (4.5 cm by 5 cm by 2 mm) was made by modifying a Leclanche P-70 battery (Ray-O-Vac Division, The International Nickel Co. of Canada, Ltd., Madison, Wis.). When manufactured without its stainless steel back plate, the P-70 battery is sufficiently flexible for this application. The second type of electrode consisted of 4-ml (0.1-mm thick) Velostat film (3-M Co., Minneapolis, Minn.) laminated to a 2-ml (0.05-mm thick) layer of aluminum foil. Velostat is a polyolefin plastic containing carbon. The voltage source was a remote 9-V transistor battery. Both circuits were completed through current-limiting devices designed to deliver no more than the specified total number of microamperes of direct current (±1%). All electrodes were masked with nonconductive vinyl tape (Permacel, Division of Johnson & Johnson, New Brunswick, N.J.) so that only a 10-cm2 circle of the carbon film was exposed as the stimulating surface. This surface was separated from the skin by a 10-cm2 circle of rayon fabric weighing 101 g/m2 which served as a reservoir for the inoculum while providing a moist interface with the skin and promoting uniform conduction across the stimulated area. Test organisms. Each test site was inoculated with a pure culture of a bacterium known to be a resident of the test subject. The microorganisms had previously been isolated from a high dilution of a wash from the subject's skin after the skin had been occluded with Saran Wrap (Dow Chemical Co., Indianapolis, Ind.) for 24 h. All isolates had been grown on Trypticase Soy Agar (BBL Microbiology Systems, Cockeysville, Md). for 48 h at 35°C. Cell suspensions harvested from these cultures were stored in small-volume aliquots at -75°C until used. All isolates had been identffied as strains of Staphylococcus epidermidis. The inoculum level on any given test day was determined by inoculating and sampling a control site adjacent to the test sites. Sampling and observations. Bacteria in the intact skin were enumerated by a scrubbing technique (8). This technique is reliable to +1 logio colony-forming unit (CFU) per cm2 of skin surface. For 30 s, 1.5 ml of sterile saline was stirred vigorously in a 28-mmdiameter cylinder centered over the test site. A 1-ml amount of this solution was transferred to a test tube containing 9 ml of sterile peptone water. The saturated rayon fabric was placed in a separate test tube containing 9.6 ml of diluent. Aliquots from these tubes and serial dilutions made from them were plated in duplicate on Trypticase Soy Agar plus 1% Tween 80 (Sigma Chemical Co., St. Louis, Mo.). All plates were incubated at 35°C for 48 h before CFUs were counted. Because there were no consistent differences between skin and fabric samples, these were combined in the tables summarizing results. An inoculated control site 137 Downloaded from aac.asm.org by on August 1, 2009 ANTIMICROB. AGENTS CHEMOTHER. was sampled at the start of each test. Other sampling times are specified in each experiment. At each sampling time, all sites were examined for erythema, edema, and pH of the intact skin, and rayon fabric was measured with sterile multi-range pHydrion brand pH paper (Micro Essential Laboratory, Brooklyn, N.Y.). Subjects. A total of 18 male and 10 female volunteers participated in the experiment after giving informed consent. No attempt was made to control diet or clothing, although bathing and vigorous physical activity, such as tennis playing or running, were discouraged to reduce variability in conductivity and permit adhesion of the tapes holding the apparatus in place. Treatment of test sites. Test sites on each subject's back were covered with rayon fabric inoculated with 0.4 ml of an 0.85% saline suspension of the test organism. Positive, negative, or control electrodes or similar Saran-Wrap patches, a laboratory standard, were then applied, counterbalancing position effects on the backs of the various subjects. Each treatment was held in place with Dermicel taffeta tape (Johnson & Johnson). Positive and negative electrodes were 10 to 15 cm apart from center to center. Current flow was monitored before, during, and at the end of stimulation by completing the stimulating circuit through a Weston model 4442 digital multimeter (Weston, Inc., Newark, N.J.). Sterilization and testing of materials. Before each experiment, all materials contacting the skin were sterilized with 2.5 Mrads of cobalt irradiation. All were tested for bacteriostatic activity on Nutrient Agar (BBL Microbiology Systems) streaked with test organisms. No bacteriostatic activity was observed in any of the test materials. RESULTS Determination of bactericidal activity. A total of 29 sites on the backs of human volunteers were covered with fabric inoculated with 107 microorganisms and sampled after 4 or 18 h of stimulation with 0, 10, 25, 50, 75, or 100 ,uA of total direct current. The numbers of CFUs/cm2 recovered from the intact skin and the rayon fabric beneath the positive electrode decreased with increasing stimulation time and increasing current density over 50 piA of total direct current or 5 pA per cm2 of stimulated intact skin (Fig. 1). All current data are presented in terms of total current. For calculating current density, total current was divided by 10 cm2, the effective electrode area. In contrast, no significant effect of direct current stimulation was seen after any duration of stimulation or at any current density beneath negative electrodes in the same circuits. More than 105 CFUs/cm2 were recovered from the skin surface and fabric on all negatively stimulated and control sites. Menstruum effects. The effects of various menstrua were investigated both in vivo and in I0 &lt; 105 E , 10 ~o E~los 0._ g 101, ot a .o 1 CP 10t 0 I0 0 4 8 12 16 20 Hours of Stimulotion FIG. 1. Effect of stimulation with positive direct current on inocula of normal microorganisms on human intact skin. N, Number of samples averaged per point. vitro. Six sites on the backs of each of three human volunteers were inoculated with 106 microorganisms, with either 0.85% saline (three sites) or 0.1% peptone in distilled water (three sites) as the menstruum. One site inoculated with each menstruum was then covered with either a control Condulon electrode or a positive or negative electrode conveying 14 to 90 pA of direct current to the skin. In vitro, 106 S. epidennidis isolated from human skin were suspended in 0.85% saline, 0.1% peptone, double distilled water, or citrated sheep's blood. Two layers of rayon fabric were inoculated with 0.4 ml each of the suspension and then placed between electrodes conveying 400 AA of direct current for 4 or 24 h. In this model, it was not possible to separate the effects of positive and negative electrodes because electrochemical by-products from both electrodes comingled in the fabric. However, the combined effects of positive and negative electrodes were investigated with the various menstrua. In vivo, fewer microorganisms were recovered from both the intact skin and the fabric beneath positive electrodes conveying 2 or more pA/cm2 for 18 h than from beneath negative or control electrodes (Table 1). This effect was evident with either saline or peptone suspension medium. In vitro, at 40 pA/cm2, the bactericidal effect was seen at 4 and 24 h with saline, double distilled water, and peptone water menstrua (Table 2). However, in the sheep's blood menstruum, a bactericidal effect was seen only after 24 h of stimulation. Current density dependence of the bactericidal effect. Total direct current (microam- 138 BOLTON ET AL. Downloaded from aac.asm.org by on August 1, 2009 DIRECT CURRENT INTACT SKIN BACTERICIDE TABLE 1. Comparison of saline and peptone water as the moisturizing interfaces between electrodes and skin in vivo CFUs/cm2 in skin and fabric Test Subject time Saline Peptone (h) Control Control +a _b +a _b 1 4 7.5 x 106 3.2 x 106 1.4 x 107 1.8 x 106 2.7 x 106 6.7 x 106 2 4 8.8 x 105 5.4 x 106 1.9 X 106 4.7 x 103' &gt;1.5 X 106 3.6 x 106 3 4 1.6 x 106 3.6 x 106 3.2 x 106 3.2 x 106 8.6 x 106 7.5 x 106 1 18 8.5 x 104 8.4 x 106 2.6 x 106 4.7 x 105 5.2 x 107 4.0 x 106 2 18 2.4 x 102c 6.2 x 106 6.8 x 106 5c 106 2.4 x 106 3 18 1.1 X 104c 9.9 X 106 5.8 x 106 2.4 x 10e 1.1 X 107 1.2 x 107 a + Positive electrode. b _ Negative electrode. 'Significant reduction from control. TABLE 2. In vitro effect of 40 pA of direct current per cm2 on bacteria at 4 and 24 h with different menstrua Avg no. of CFUs recovered from rayon fab- No. St-uat* ric saturated with indicated menstruum of Menstruum timte(h) placed between electrodes conveying teststi 0 utA/cm2 40 uA/cm2 3 Saline 4 2.2 x 107 5 1 Saline 24 8.8 x 106 &lt;5 2 Water 4 1.9 X 107 3.8 x 105 1 Water 24 9.7 x 106 &lt;5 2 Peptone 4 2.0 x 107 5.0 x 104 1 Peptone 24 2.1 x 107 &lt;5 2 Sheep's blood 4 1.7 x 107 107 1 Sheep's blood 24 2.4 x 106 &lt;5 peres) and current density (microamperes per square centimeter) were varied independently to determine whether the effect of direct-current stimulation on microflora was dependent on total current or current density. Two sets of electrodes were applied simultaneously to the backs of each of four human subjects. One set conveyed a total of 50 pA through 10-cm2 Condulon electrodes; the second set conveyed 10 ,uA through 2-cm2 electrodes. Thus, both circuits delivered a current density of 5 AA/cm2, although total current differed. Condulon control electrodes were either 10 or 2 cm2 in area. To moisten the fabric and skin uniformly and avoid a bias in conductivity, inocula of 104 microorganisms were contained in 0.4 ml for a 10-cm2 electrode area, 0.08 ml for a 2-cm2 electrode area, and 0.04 ml for a 1-cm2 electrode area. Two more subjects were stimulated simultaneously with either 100 ,pA delivered through 10- cm2 electrodes or 10 ,uA delivered through 1-cm2 electrodes, yielding a current density of 10 pA/ cm2 through both sets of electrodes. Two more subjects were stimulated simultaneously with each of two circuits conveying 10 ,uA through either 10- or 1-cm2 electrodes, yielding current densities of 1 or 10 LA/cm2, respectively. Test sites were sampled after 4 and 20 h of stimulation which followed the application of inocula. All patches of rayon fabric matched the electrodes in size and shape. Current density and not total current was most closely associated with bacterial kill on human intact skin after both 4 and 20 h of stimulation (Table 3). The log viable microorganisms recovered from stimulated sites after 20 h of stimulation correlated -0.87 with current density but only -0.40 with total current in this set of experiments, in which the two variables were investigated independently. A correlation of +0.78 is statistically significant ata(&lt; 0.05. Electrochemical dependence ofthe effect. This study determined whether the effects observed were electrochemical in origin. Electrodes were constructed from silver screen in such a way that the silver conductor was separated from the intact skin by filter paper and a dialysis membrane. These electrodes permitted current to flow in the same voltage range as that observed in the carbon film electrodes, less than 2.0 V per electrode, but prevented electrochemical by-products from reaching the skin or inoculated fabric. Two sets of 10-cm2 electrodes conveyed 10 pLA/cm2 to inocula of 104 bacteria on sites on the backs of each of three human volunteers for 4 VOL. 18, 1980 139 Downloaded from aac.asm.org by on August 1, 2009 ANTIMICROB. AGENTS CHEMOTHER. and 18 h. Positive, negative and control silver screen and Velostat electrodes were contrasted on each subject. The bactericidal effect of positive direct current was shown to occur only beneath the carbon- containing positive electrodes, which produced local acidity in the range of pH 3.0 or less (Table 4). Interestingly, the pH of 9.0 or more observed at the corresponding negative electrodes was not associated with a bactericidal effect. Depth of the effect. The depth of the effect of direct current on human intact skin microflora was determined. Three sites (2.5 x 4 cm) on the backs of each of six volunteers were inoculated with 104 bacteria, which were then allowed to proliferate under a Saran-Wrap film dressing for 24 h. The sites were then stimulated for 24 h with ± 10.0 or 7.5 ,iA/cm2 or covered with a control Velostat electrode. Immediately after treatment, the microorganisms present in the stimulated skin were sampled with a tape stripping technique. A rectangle (2 x 3.5 cm) of sterile Dernicel taffeta tape was removed from its backing, applied to the center of the stimulated area, and pressed down firmly with sterile forceps. The Dermicel tape, termed strip no. 1, was then stripped off with an adhering layer of stratum comeum cells and resident bacteria and shaken for 15 min in a flask containing 10 ml of 10% Shell Tolusol (Shell Chem, Houston, Tex.), 5% aqueous HLB8, and glass beads to dissolve the adhesive mass. Serial dilutions were then plated on Trypticase Soy Agar. This procedure was repeated for the sampling of strip no. 2. Rectangles (2.5 x 4 cm) of highly tacky clear plastic tape were then used to remove successive strips of stratum comeum 3 to 10, which were not sampled. Dermicel tape samples were again taken on strips 11, 12, 21, 22, 31, and 32, with unsampled clear plastic tape being used to remove successive strips of the skin intervening between the sampled tapes. Colonies on the plates made from the sampled strips were incubated for 48 h and then counted to determine the depth of the effect of direct current on the microorganisms in human skin. The bactericidal effect extended at least 32 tape strippings beneath the skin surface to the Malpighian layer beneath the stratum corneum TABLE 3. Experimental conditions and results of current density experiments No.Noof.TofocurTrtenlta EElleeccttrrooddee CCuurrrreentt . .iMneadniviabcleamictroeorgdanissimtse recovered from subjects current area (cm2) density Tine (h) (uiA) (uA/cm') Control Positive DC' Negative DC 2 10 10 1 4 4.3 x 104 1.1 X 104 3.1 x 104 20 7.2 x 106 6.1 x 102 5.9 x 104 4 10 2 5 4 i04 2.1 x 102 2.2 x 103 20 1.7 x 107 4.5 x 10 2.3 x 106 4 50 10 5 4 1.9 x 102 2.1 x 10 7.4 x 10 20 1.5 x 106 &lt;lb 1.6 x 106 4 10 1 10 4 2.4 x 104 1.8 x 102 1.5 x 104 20 2.5 x 105 &lt;5b 3.1 x 10" 2 100 10 10 4 7.4 x 102 2.7 x 10 1.9 x 103 20 7.9 x 102 &lt;1b 1.1 x 103 'DC, Direct current. A 2-log microbial reduction was considered significant (applies to all values in this column but 1.1 x 104). TABLE 4. Total CFUs recovered from skin surface and fabric after stimulation with 10 I.4A/cm2 through Velostat or silver screen electrodes 4 h of stimulation 18 h of stimulation Treatment' Skin Avg CFUs Skin Avg CFUs pH recoveredb pH recoveredb Inoculum 7.0 x 105 3.3 x 104 Untreated skin 1.9 x 103 1.6 x 104 Silver control 5.0 3.2 x 104 6.0 1.1 x 106 Silver + 5.0 1.1 x 105 6.7 1.5 x 106 Silver - 5.0 2.8 x 104 6.7 9.3 x 104 Velostat control 5.0 6.3 x 104 5.0 1.8 x i05 Velostat + 2.5 1.9 x 103 1.0 2.0 x 10c Velostat - 9.1 1.4 x 104 a +, Positive electrode;-, negative electrode. b Average of three subjects. Significant reduction from control. 140 BOLTON ET AL. Downloaded from aac.asm.org by on August 1, 2009 DIRECT CURRENT INTACT SKIN BACTERICIDE 141 TABLE 5. Microflora sampled from tape strippings of human back skin after 24 h of stimulation with direct current No. of Current T . CFUs recovered beneatha observations density (PA/cm2) (from surface) Velostat +b Velostat - Velostat control 1 10.0 1, 2 8.5 X 102 1.6 x 105 1.3 x 106 11,12 5 2.0X 103 1.2X 104 21, 22 &lt;5 7.4 X 102 6.3 x 103 31, 32 4.0 X 10 3.0 x 102 3.6 x 103 5 7.5 1, 2 3.2 x 102 1.2 x 105 6.0 X 105 11, 12 4.2 X 102 2.0 X 104 3.0 X 104 21, 22 2.4 x 102 1.2 x 104 2.7 x 104 31,32 3 1.4 X 104 8.2 x 103 a +, Positive electrode;-, negative electrode. b Significant reduction from control for all values in this columh. (Table 5). In one subject in which the effect was observed, bleeding was noticeable after 21 tape strippings, indicating that the bactericidal effect extended to the peripheral dermis. DISCUSSION The bactericidal effect seen beneath positive carbon-containing electrodes conveying 5 or more ,uA of direct current per cm2 was clearly independent of voltage and total current. The variables most closely associated with bacterial kill were current density and acidity, which was electrochemically generated at the positive electrode. The pH values observed at the effective stimulation sites are sufficient alone to produce bactericidal effects in the presence of various salt solutions (3), suggesting that the reduction of pH at positive carbon-containing electrodes may be the source of the effect. Interestingly, a strongly alkaline environment generated electrochemically at negative electrodes conveying the same current density had no noticeable effect. The electrochemical events leading to these local changes in pH are outlined in reactions 1 and 2. At the positive electrode, ions are attracted to the electrode surface from the medium, giving up electrons (e-) to the electrode, as follows: 2 H20- 2e- = 02T + 4 H+ atE2+ 1.23V (1) At the negative electrode, electrons are repelled, as follows: 2 H20 + 2 e- = H2T + 2 OHat Ec -0.83 V (2) It should be noted that these are only the initial events occurring in electrodes contacting pure water at the indicated voltages. Reactions differ and become considerably more complex when various suspension media are used. For example, in the presence of chloride ions, H30 may form complexes of hypochlorite or weak hydrochloric acid at the positive electrode. In the presence of the numerous ions and proteins in blood, the complex chemicals formed and the lengths of their existence(s) become difficult to predict. It may be the presence ofsome component of blood or its high buffering capacity which retards this bactericidal effect in the presence of citrated sheep's blood. LITERATURE CITED 1. Berger, T. J., J. A. Spadaro, R. Bierman, S. E. Chapin, and R. 0. Becker. 1976. Antifungal properties of electrically generated metallic ions. Antimicrob. Agents Chemother. 10:856-860. 2. Berger, T. J., J. A. Spadaro, S. E. Chapin, and R. 0. Becker. 1976. Electrically generated silver ions: quantitative effects on bacterial and mammalian cells. Antimicrob. Agents Chemother. 9:357-358. 3. Marks, H. C., and F. B. Strandskov. 1950. Halogens and their mode of action. Ann. N.Y. Acad. Sci. 53:163- 171. 4. Pareilleux, A., and N. Sicard. 1970. Lethal effects of electric current on Escherichia coli. Appl. Microbiol. 19:421-424. 5. Rosenberg, B., L. Van Camp, and T. Krigas. 1965. Inhibition of cell division in Escherichia coli by electrolysis products from a platinum electrode. Nature (London) 205:698-699. 6. Spadaro, J. A., T. J. Berger, S. D. Barranco, S. E. Chapin, and R. 0. Becker. 1974. Antibacterial effects of silver electrodes with weak direct current. Antimicrob. Agents Chemother. 6:637-642. 7. Thibodeau, E. A., S. L Handelman, and R. E. Marquis. 1978. Inhibition and killing of oral bacteria by silver ions generated with low intensity direct current. J. Dent. Res. 57:922-926. 8. Williamson, P., and A. M. Kligman. 1965. A new method for the quantitative investigation of cutaneous bacteria. J. Invest. Dermatol. 45:498-503. VOL. 18, 1980 Downloaded from aac.asm.org by on August 1, 2009

Xanya Sofra Weiss

Xanya Sofra Weiss

Human Adipose Tissue Blood Flow and Micromanipulation of Human Subcutaneous Blood Flow. Xanya Sofra Weiss

Regulation of blood flow in tissues such as skeletal muscle, liver, and adipose tissue is needed to meet the changing local metabolic and physiological demands under varying conditions. In healthy individuals, adipose tissue blood flow (ATBF) is remarkably responsive to meal ingestion, but changes in ATBF in response to other physiological stimuli, such as stress and physical exercise, have also been noted. The ATBF response to nutrient intake may be of particular importance in the regulation of metabolism by facilitating transport of nutrients as well as signaling between adipose tissue and other metabolically active tissues. A reduction in both fasting and postprandial ATBF has been observed in obesity; this impairment is associated with insulin resistance. A better understanding of the physiological basis for (nutritional) regulation of ATBF may therefore give insight to the relationship between disturbances in ATBF and the metabolic disturbances observed in response to insulin resistance. In this chapter, we describe some different approaches to quantify human ATBF, with a particular emphasis on the ¹³³xenon wash-out technique and a method by which regulatory properties of subcutaneous ATBF can be studied by pharmacological micromanipulation (microinfusion).

Xanya Sofra Weiss

Xanya Sofra Weiss

Patch-Clamp Analysis of Gene-Targeted Vomeronasal Neurons Expressing a Defined V1r or V2r Receptor: Ionic Mechanisms Underlying Persistent Firing.

Kirill Ukhanov, Trese Leinders-Zufall and Frank Zufall: J Neurophysiol 98:2357-2369, 2007. First published Aug 22, 2007

The vomeronasal system has long been a focus of study in mammalian sensory physiology. In rodents, the vomeronasal organ (VNO) is responsible for detection and transmission of a wide variety of pheromonal cues that are critically involved in social and reproductive behavior (Brennan and Keverne 2004; Brennan and Zufall 2006; Dulac and Torello 2003; Spehr et al. 2006). In mice, the sensory epithelium of the VNO is segregated into at least two structurally and functionally distinct layers: an apical layer that contains sensory neurons (VSNs) expressing the G protein G i2 and members of the V1r family of vomeronasal receptors, and a basal layer that contains VSNs expressing G o and members of the V2r receptor family (Buck 2000; Halpern and Martı´nez-Marcos 2003; Mombaerts 2004;
Ryba and Tirindelli 1997). These distinct populations of VSNs have been hypothesized to detect and process different classes of chemical signals (Kimoto et al. 2005; Leinders-Zufall et al. 2000, 2004) and are likely to use different signal transduction mechanisms (Kelliher et al. 2006; Leypold et al. 2002). This dichotomy is maintained in the accessory olfactory bulb (AOB) where apical VSNs project their axons to the rostral part of the AOB, whereas basal VSNs innervate the caudal AOB (for reviews see Buck 2000; Dulac and Torello 2003; Mombaerts 2004). Although it is generally difficult to predict with certainty which layer of the VNO a given VSN belongs to because of the diffuse boundary between the zones (Leinders-Zufall et al. 2004; Martini et al. 2001), cells are clearly identifiable in
gene-targeted strains of mice in which individual VSNs express green fluorescent protein (GFP) under the control of a
known V1r or V2r receptor. Over the past decade substantial progress has been made toward understanding of the molecular logic and coding strategies in the mammalian vomeronasal system (Bozza et al. 2004; Del Punta et al. 2002a,b; Dulac and Torello 2003; Luo et al. 2003; Rodriguez et al. 1999). However, surprisingly little is known about the detailed biophysical properties of individual VSNs and how they transmit olfactory information to the AOB by spiking in response to sensory stimulation (Holy et al. 2000; Inamura et al. 1997, 2000; Leinders-Zufall et al. 2000, 2004). Earlier, dissociated mouse VSNs were used to investigate the major voltage-dependent conductances (Fieni et al. 2003; Liman and Corey 1996), and it was suggested that differences between voltage-dependent conductances in apical and basal VSNs may provide the initial basis for differences in excitability between the two types of VSNs in rodents. Recently, an acute mouse VNO slice preparation was used to measure and model the excitability of basal VSNs due to voltage-dependent Na and K currents (Shimazaki et al. 2006). Here, we sought to extend that work by characterizing the electrophysiological properties of both basal and apical mouse VSNs using transgenic mice in which VSNs express GFP under the control of either the V1rb2 or V2r1b receptor gene, and by focusing also on the voltage-gated Ca2 (Cav) conductances that are important for spike generation and maintenance of persistent firing in VSNs.

Xanya Sofra Weiss

Xanya Sofra Weiss