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The Effects of Madol on the Physical Activity - Research Paper Example

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The paper describes the knowledge of the effects of Madol on the physical activity that was forthcoming, the present studies were designed to study the psychopharmacological effects of Madol on the activity of hamsters using a locomotor activity assay based on photocell technology…
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The Effects of Madol on the Physical Activity
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Effect of Madol, a designer steroid, on the activity of hamsters. Rationale: Anabolic androgenic steroids are man made drugs resembling the male sex hormone function and structure. These drugs are muscle building (anabolic) and promoters of masculine characteristics (androgenic) and are frequently abused by athletes to enhance their performance. Madol, chemically known as Desoxymethyl-Testosterone (DMT), is a latest designer steroid, developed to avoid detection in the urine sample analysis of the athletes. Madol has been characterized chemically and structurally and assays are now available to detect the presence of Madol in urine samples (Sekera, 2005). Since, the knowledge of the effects of Madol on the physical activity was forthcoming, the present studies were designed to study the psychopharmacological effects of Madol on the activity of hamsters using a locomotor activity assay based on photocell technology. Hypothesis: Based on the aforementioned reasoning, it is hypothesized that "Madol (DMT) induces the activity levels of hamsters as assessed by locomoter activity." Experimental design: One of the most basic experimental designs used in psychopharmacology is to study the effect of drugs on the locomotor activity of the animals. This experimental design is often used as an initial screen for pharmacological effects of a drug class in humans on psychological/neuronal function. In the study, measurement of the activity of hamsters will be assessed using the locomotor activity with photocell-based automated monitoring system. Photocell is a solid-state device that has a light sensitive cathode and releases electrons to anode upon illumination. This current (flow of electrons) is measured and is proportional to the intensity of the illumination from infrared rays (Pierce, 2007). Animals and Housing Conditions: For the purpose of this study 3-month-old adult Hamsters (Mesocricetus auratus) will be reared in one climate controlled room in standard plastic cages. Standard food pellets and water will be provided ad libitum. The light/dark conditions will be 12:12 with the light intensity varying between 200 to 300 Lx (Unit of luminance) during day and 5 Lx (Unit of luminance) during the dark period. Temperature will be regulated to 25 + 2 Celsius and the relative humidity will be maintained between 60 to 65 %. The protocols for using the hamsters for the experiments will be reviewed and approved by an Institutional Animal Care and Use Committee (IACUC) and all National Institute of Health (NIH) guidelines and procedures for care and use of laboratory animals will be followed. Drug treatment: Hamsters (n = 80) will be divided into four groups. Group I (n = 8) will receive an intraperitoneal injection of equivalent volume of ethanol used in other groups for drug delivery. Group II (n = 24, 8 per each dose) will be injected subcutaneously with testosterone (1,2 or 5 mg/Kg body weight). Group III (n = 24, 8 per each dose) will be injected intraperitoneally with dihydrotestosterone (DHT, 1,2, or 5mg/Kg body weight). Group IV (n = 24, 8 per each dose) will be treated with madol (DMT, 1,2, or 5 mg/Kg body weight) intraperitoneally. Testosterone, DHT (derived from testosterone by enzyme 5-alpha reductase) and DMT will be dissolved in ethanol. Testosterone and DHT are natural anabolic androgenic steroids present in human body. Dose levels are derived from extensive literature review for testosterone and DHT. All these drugs will be procured from a chemical company. Locomotor activity quantification by photocell-based automated monitoring system: Several photocell-based behavioral apparatus are commercially available. These apparatus have an approximately 40 40 cubic cm arena. Horizontal activity can be measured by placing the photocells (photo sensors) at a distance of 2.5 cm along the x- and y-axis. These photocells measure the intensity of the invisible grid (of infrared light) placed at 4 cm (lower level) or 12.5 cm (upper level) above the floor of the activity arena. Any disruption of the intensity measurement represents the activity of hamsters. Placement of these photocells at two different levels could be used to measure different activities described in experiment section below. The whole arena is placed in a box, which is sound, attenuating and ventilated (Pierce, 2007). Preparation of the activity apparatus and subject: Activity arena will be cleaned with soap water followed by alcohol after each experiment. Before using the apparatus, the alcohol is allowed to evaporate. Computers and the interfaces to monitor the activity will be set up according the manufacturer's instructions. The animal subject will be allowed to habituate for 60 minutes hours to minimize new environment induced freezing of exploratory behavior and injected with vehicle (ethanol). The animal will be allowed to return to arena for further habituation for 2 hours and returned to its regular cage. Experiment: After 24 hours of habituation and injections, the subject will be placed in a clean apparatus in the activity arena. Computer will be allowed to record the locomotor activity of the subject for 60 minutes. The animals will be injected with vehicle (ethanol) or the drugs depending on their group and dosage. The computer will record locomotor activity for two hours. The time course may be adjusted depending on the dosage of the drugs used in this study. The data generated from this experiment, including motor activity (all interruptions of the photo beams), peripheral (limb) motor activity as detected by interruptions in lower beam provided beams are 25 mm apart, rearing (rise on hind limbs) represented by interruptions in upper beam, peripheral rearing as seen by interruption of upper beams spaced 25 mm apart, locomotion shown by successive disruptions of photo beams in the lower level, speed demonstrated by time between consecutive photo beam interruptions during locomotion will be analyzed (Pierce, 2007). All these activities will be subsequently monitored for a week, followed by measurements at 2 weeks and 1 month to study the long-term effects. Statistical Analysis: Beam crossings as detected by photocell will be recorded every minute and the average crossings for each group of hamsters summed into 20-min intervals for graphical display in each session. Average activity for each of the activity assessed (motor activity, peripheral activity, rearing, peripheral rearing, locomotion and speed) by beam crossing per session will be analyzed over a period of time to assess the long-term effects of Madol on activity. Statistical analysis of the individual sessions will be performed by means of a one-way ANOVA, followed by Dunnett's t-test for comparison with the vehicle control or comparisons between different groups. Statistical evaluation between different treatment groups (variable 1) over a period of time (variable 2) will be carried out using a two-way ANOVA for repeated measurements (Winer, 1971). Grand mean for each group will be calculated and the difference considered statistically significant if the p value is less that 0.05. Advantages of photocell-based systems: Although locomotor activity can be assessed by using a variety of experimental designs including direct observation using scales, video-based systems, rotometers, and running wheels, photocell-based system provides distinct advantages, as it allows complex locomotor behavior described above, to be analyzed. Only drawback of these systems is their expense. Total package including the computers, data controllers, and behavioral arenas (eight) range in price from $30,000 to $50,000. Expected results and alternatives: Using the automated photocell-based system we expect the activity levels of hamsters treated with madol to be significantly higher compared to that of vehicle, testosterone and DHT. Since this drug of abuse is not characterized in any functional activity studies, the adverse reaction it may cause on psychopharmacological profile as assessed by locomoter activity could be documented. References Pierce, R.J., and Kalivas, P.W. (2007) Locomotor behavior. Wiley-Interscience. Sekera, M.H., Ahrens, B.D., Chang, Y.C., Starcevic, B., Georgakopoulos, C., and Catlin, D.H. (2005). Another designer steroid: discovery, synthesis, and detection of 'madol' in urine. Rapid Communication in Mass Spectrometry, 19, 781-784. Winer, B.J. (1971) Statistical principles in experimental design. McGraw-Hill Companies. Read More
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