Nukleus Akumbens Bölgesine Uygulanan Topiramatın Morfin Yoksunluk Sendromuna Etkileri

Aim: Nucleus accumbens, one of the nuclei of the basal ganglia, and dopamine, the neurotransmitter play a critical role in opioiddependence and withdrawal. In opioid withdrawal, the importance of neurotransmitters such as glutamate and gamma aminobutyric acid(GABA), as well as dopamine, is known. In this study, we aimed to investigate the effects of local injections of topiramate, anantiepileptic agent affecting GABAergic and glutamatergic pathways, into the nucleus accumbens on withdrawal signs and locomotoractivity during naloxone-induced withdrawal in morphine-dependent rats.Materials and Methods: Twenty male Sprague-Dawley rats were divided in topiramate treatment and control groups. All animalsreceived morphine pellets and guide cannulas were placed bilaterally in the nucleus accumbens regions by stereotaxic surgery. On thelast day of the experiment, following the bilateral topiramate or saline (control group) microinjections, morphine withdrawal was triggeredby naloxone.Results: Topiramate microinjections into the nucleus accumbens region significantly suppressed the signs of naloxone-inducedmorphine withdrawal such as number of jumpings and weight loss. No significant difference was observed in wet dog shakes, one of thewithdrawal signs, after local topiramate treatment. Although topiramate microinjections increased stereotypical activity it did not changelocomotor activity behavior such as vertical and ambulatory activity, and total covered distance.Conclusion: These findings show that local microinjection of topiramate into the nucleus accumbens is effective in preventing opioiddeprivation symptoms without significant effect on locomotor activity.Amaç: Bazal gangliyon çekirdeklerinden biri olan nukleus akumbens ve nörotransmitter olan dopamin opioid bağımlılığı ve yoksunluğunda kritik rol oynamaktadır. Opioid yoksunluğunda dopaminin yanı sıra glutamat ve GABA gibi nörotransmitterlerin de önemi bilinmektedir. Biz bu çalışmada morfin bağımlılığı oluşturulan hayvanlarda GABAerjik ve glutamaterjik yolakları etkileyen antiepileptik ajan olan topiramatın nukleus akumbens bölgesine lokal uygulamasının naloksonla tetiklenen yoksunluk sendromunda yoksunluk bulguları ve lokomotor aktivite üzerine etkilerini araştırmayı amaçladık. Materyal ve Metot: Yirmi adet erkek Sprague-Dawley sıçanları topiramat tedavi grubu ve kontrol grubu olarak ikiye ayrıldı. Hayvanların hepsine morfin peletleri uygulandı, stereotaksik cerrahi işlemle nukleus akumbens bölgelerine kılavuz kanüller bilaretal yerleştirildi. Deneyin son gününde bilateral topiramat veya serum fizyolojik (kontrol grubu) mikroenjeksiyonlarını takiben nalokson uygulanarak morfin yoksunluğu tetiklendi. Bulgular: Nukleus akumbens bölgesine lokal uygulanan topiramat naloksonla tetiklenen morfin yoksunluk bulgularından sıçrama sayısını ve ağırlık kaybını anlamlı düzeyde baskıladı. Lokal topiramat uygulaması yoksunluk bulgularından ıslak köpek silkinmesinde ise anlamlı değişiklik yapmadı. Topiramat mikroenjeksiyonları stereotipik hareketleri artırdığı halde vertikal hareketler, ambulatuvar hareketler ve toplam kat edilen mesafe gibi lokomotor aktivite davranışlarını değiştirmedi. Sonuç: Bu bulgular antikonvülzan ilaç olan topiramatın nukleus akumbens bölgesine lokal uygulanmasının lokomotor aktivitede anlamlı baskılanma yapmadan opioid yoksunluk belirtilerinin önlenmesinde etkili olduğunu göstermektedir

The Effects of Topiramate Applied to the Nucleus Accumbens Region on Morphine Withdrawal Syndrome

Aim:Nucleus accumbens, one of the nuclei of the basal ganglia, and dopamine, the neurotransmitter play a critical role in opioid dependence and withdrawal. In opioid withdrawal, the importance of neurotransmitters such as glutamate and gamma aminobutyric acid (GABA), as well as dopamine, is known. In this study, we aimed to investigate the effects of local injections of topiramate, an antiepileptic agent affecting GABAergic and glutamatergic pathways, into the nucleus accumbens on withdrawal signs and locomotor activity during naloxone-induced withdrawal in morphine-dependent rats.Materials and Methods:Twenty male Sprague-Dawley rats were divided in topiramate treatment and control groups. All animals received morphine pellets and guide cannulas were placed bilaterally in the nucleus accumbens regions by stereotaxic surgery. On the last day of the experiment, following the bilateral topiramate or saline (control group) microinjections, morphine withdrawal was triggered by naloxone.Results:Topiramate microinjections into the nucleus accumbens region significantly suppressed the signs of naloxone-induced morphine withdrawal such as number of jumpings and weight loss. No significant difference was observed in wet dog shakes, one of the withdrawal signs, after local topiramate treatment. Although topiramate microinjections increased stereotypical activity it did not change locomotor activity behavior such as vertical and ambulatory activity, and total covered distance.Conclusion:These findings show that local microinjection of topiramate into the nucleus accumbens is effective in preventing opioid deprivation symptoms without significant effect on locomotor activity

Evaluation of alcohol and psychoactive substances in blood samples of drivers involved in traffic accidents

Objectives: Traffic accidents resulting from driving under the influence of drugs (DUID) are a serious problem throughout the world. This study evaluated the type of the psychoactive substances (with or without alcohol) detected in blood samples of drivers involved in traffic accidents in Istanbul and surrounding area. Materials and methods: The samples from individuals involved in traffic accidents were sent for analysis to the Council of Forensic Medicine-Istanbul Chemistry Department by police or judicial authorities between July 2010 and June 2011. Alcohol analysis was carried out by headspace HS/GC Perkin Elmer Clarus 500 GC TMHS 40. Psychoactive substances were detected using an immunoassay screening (Microgenics’ CEDIA Assay kits including Amphetamine/Ecstasy, Benzodiazepines, Barbiturates, Multilevel THC, Cocaine, Methadone, Opiate, TCA Antidepressants on Hitachi 911) followed by LC/MS/MS. Blood samples were subjected to solid phase extraction using Oaisis HLB(60 mg) cartridges. Cartridges were conditioned with 1 ml methanol and water respectively. Then 1.5ml blood was passed. Cartridge rinsed by 5% Methanol in water (v:v) and dried. And finally eluted 0.5 ml methanol, 0.5 ml 2% ammonia in methanol (v:v) and 0.5 ml 2% Acetic Acid in methanol(v:v) respectively. Analyses were performed on a Shimadzu LC-20A series system interfaced to a tandem mass spectrometry API 4000 with an electrospray Turbo V™ ion source in negative and positive mode. For positive mode the ESI source settings were ion spray voltage 5500 V, source temperature 500 °C, nebulation and heating gas (N2), 50 and 50 respectively. For negative mode the ESI source settings were ion spray voltage -4500 V, source temperature 500 °C, nebulation and heating gas (N2), 50 and 50 respectively. For chromatographic separation polar full endcapped phenylpropyl column was used at 40 °C. A mobile phase of water containing 0.2% formic acid and 2 mM Ammonium Formate (Solvent A) and acetonitrile containing 0.2% formic acid and 2 mM Ammonium Formate (Solvent B) was delivered with the following gradient: starting 10% B at 1ml/min, reaching 90% B in 10 min at 1ml/min, for 5 min 90% B at 1ml/min, back to 10% B in 2.5 min at 0.5 ml/min. Detection of the ions was performed in multiple reaction monitoring (MRM) mode. Analysis of the collected data was carried out by means of Analyst software (Applied Biosystems/Sciex). Results: Alcohol analysis was requested in 4274 traffic accident cases by legal authorities during this 1 yr period. However, the analysis of psychoactive substances along with alcohol was requested for only 92 cases. Blood analysis in 32 (35.1%) of these 92 suspected cases showed only alcohol; however, in 14 (15.2%) of cases at least one psychoactive substance was identified. There was neither alcohol nor psychoactive substances detected in 45 (49.4%) cases. Out of these 14 cases, in 8 (57.1%) both alcohol and psychoactive substances were identified and in 6 (42.8%) cases there was at least one psychoactive substance without alcohol. THC was the most frequent substance detected (n=8; 57.1%). Benzodiazepines ((n=4; midazolam (n=3) and diazepam (n=1)), pentobarbital (n=2), MDMA (n=1), and cocaine (n=1) were the other identified psychoactive substances in these blood samples. Conclusions: Analysis for both psychoactive substances and alcohol was requested in only 1.7% of traffic accident cases during the 1 yr investigation period, and fifteen percent of them contained at least one psychoactive substance with or without alcohol. These results indicate that although there are regulations, DUID is not well recognized and not inspected by police and legal authorities who are responsible in routine controls in Istanbul, Turkey. Psychoactive substances should be checked in all traffic accident cases along with alcohol

Trafik kazası geçirmiş sürücülerde kan alkol ve psikoaktif madde analizlerinin değerlendirilmesi

Her yıl dünyada gittikçe artan sayıda trafik kazası meydana gelmektedir. Kaza sebeplerinin en önemlilerinden birini sürücülerin güvenli sürüşünü etkileyen madde kullanımları oluşturmaktadır. Çalışmamızın amacı; trafik kazaları akabinde laboratuvarımıza gönderilen sürücü kanlarında hangi psikoaktif maddelerin (uyuşturucu/uyarıcı maddeler, psikoaktif ilaç etkin maddeleri) bulunduğunu, bulunma sıklıklarını ve alkolle birliktelik oranlarını tespit etmektir. Yöntem: Adli Tıp Kurumu Kimya İhtisas Dairesi'ne Temmuz 2010-Temmuz 2011 süreci içerisinde gönderilen ve alkolle birlikte psikoaktif madde taraması istenen trafik kazası geçirmiş sürücü kanlarındaki analiz sonuçları değerlendirmeye alınmıştır. Alkol analizleri, Headspace GC-FID cihazında çalışılmış, psikoaktif madde taramaları ise immunoassay yöntemlerle analiz edildikten sonra LC/MS/MS ile teyid edilmiştir. Bulgular: 1 yıllık süre zarfında ilgili konuda yapılan araştırmada 4274 vakada trafik kazası nedeni ile alkol analizi istenmiştir. Bu vakalar içerisinde sadece 91'inde alkol ile birlikte psikoaktif madde analizi talep edilmiştir. Doksanbir vakadan 32 (%35,1) tanesinde sadece alkol, 6 (%6,6) tanesinde psikoaktif madde, 8 (%8,8) tanesinde alkol ve psikoaktif maddelerinin birlikte bulunduğu, 45 (%49,4) numunede ise aranılan maddelerin bulunmadığı tespit edilmiştir. Bulunan uyuşturucu/uyarıcı maddeleri THC, kokain ve MDMA oluştururken, psikoaktif ilaç etkin maddeleri benzodiazepin ve barbitürat türevleridir. Sonuç: Trafik kazası yapmış olan sürücülerin yaklaşık %15'inin kanlarında alkolle birlikte veya tek başına psikoaktif madde tespit edilmiştir. Ülkemizde trafik kazaları sonucu ya da rutin trafik denetimleri sırasında sürücülerin madde etkisi altında olup olmadıkları hususunda genellikle sadece alkol düzeyleri baz alınmakta, bunun için alkolmetre cihazıyla solunum havasından veya olay sonrası alınan kanlarda alkol analizleri yapılmaktadır. Halbuki, alkolden başka diğer birçok psikoaktif maddenin de sürüş emniyetine olumsuz yönde katkısı olduğu bilinmekte, trafikteki herhangi bir ölçüm sırasında diğer maddelerin tespit edilmeyeceğini bilen sürücülerin de tercihi diğer maddeler olabilmektedir. Bu nedenle rutin trafik kontrolleri ve kazalar sonucu sadece alkol analizi ile yetinilmemesi gerektiği kanaatindeyiz

Effect of brain acetylcholine depletion on bicuculline-induced cardiovascular and locomotor responses

Both GABAergic and cholinergic systems are involved in central cardiovascular regulation. Previous studies have shown that GABA(A) receptor antagonists cause increases in blood pressure, heart rate and locomotor activity. In this study, we examined the role of the depletion of brain acetylcholine on the cardiovascular responses and locomotor activity induced by bicuculline methiodide in conscious Sprague-Dawley rats. The doses of 0.3 and 0.5 nmol of intracerebroventricular bicuculline methiodide produced increases in blood pressure, heart rate and locomotor activity. The dose of 18 nmol of hemicholinium-3 to deplete brain acetylcholine was given intracerebroventricularly one hour prior to bicuculline methiodide. The presser responses to bicuculline methiodide in animals pretreated with the hemicholinium-3 were higher than those seen in saline-pretreated groups, bur locomotor activity and heart rate responses to bicuculline methiodide remained unchanged in hemicholinium-3 pretreatment group. On the other hand, high dose of bicuculline methiodide (0.5 nmol) caused convulsions in some animals pretreated with hemicholinium-3 whereas bicuculline methiodide, alone, did nor cause any seizure activity. In conclusion, it seems likely that endogenous brain acetylcholine could be a modulator of GABA(A) receptor-mediated blood pressure control

A review of suspected cases of driving under the influence of drugs (DUID) involved in traffic accidents in Istanbul (Turkey)

Nowadays traffic accidents, which have high mortality and morbidity, are an important public health problem. The association between the use of alcohol and/or drugs by drivers and the increased risk of traffic accidents with a high risk of death and injury has been well described in the literature. This study aimed to review the incidence of cases of driving under the influence of drugs (DUID) among all cases of driving under the influence (DUI) of alcohol and/or other drugs involved in traffic accidents and to evaluate the type of the psychoactive drugs (with or without alcohol) detected in blood samples in Istanbul and its surrounding area. This study is the first investigation on the subject of DUID cases in Turkey. The reports of the Istanbul Toxicology Department of the Council of Forensic Medicine (Turkey) on suspected DUID cases involved in traffic accidents between 1 July 2010 and 30 June 2011 were retrospectively reviewed for alcohol and/or drug use. Alcohol analysis was requested in 4274 suspected DUI cases, whereas drug along with alcohol analysis was requested in only 91. The rate of suspected DUID cases (n = 91) among the suspected DUI cases (n = 4274) was only 2.1% and in this study, we evaluated only the DUID cases in detail. Alcohol was present in 44% of suspected DUID cases. Psychoactive drugs were present in 15.4% of cases. The incidence among 46 confirmed DUID cases was found to be 17.4% for cannabis, 8.7% for benzodiazepines, 4.3% for barbiturates, 4.3% for antidepressants, 2.2% for cocaine and 2.2% for amphetamines. Although there is a zero-tolerance approach for DUID in the Turkish regulations, it is not well recognised and not inspected by police and legal authorities who are responsible for taking measures in traffic accidents and for routine traffic controls in Turkey. It is concluded that psychoactive drugs should be checked as well as alcohol in all traffic accident cases and roadside controls. (c) 2013 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved

Review of detection frequency and type of synthetic cannabinoids in herbal compounds analyzed by Istanbul Narcotic Department of the Council of Forensic Medicine, Turkey

In recent years, synthetic cannabinoids have been frequently observed in seized materials all over the world. This new generation of designer drugs, mixed with herbal substances, is also known as Herbal Highs or Legal Highs. There are many articles about the history, type and pharmaco-chemical properties of synthetic cannabinoids in the literature; however the number of articles about the frequency of their detection is limited. In this study, we evaluated the type and detection frequency of synthetic cannabinoids in Istanbul and its surrounding area. The reports of the Council of Forensic Medicine-Istanbul Narcotic Department were retrospectively reviewed for the presence of synthetic cannabinoids in herbal compounds sent by the judicial authorities between August 01, 2010 and March 31, 2012. Among 1200 herbal compounds, 1179 of them (98.3%) contained synthetic cannabinoids. Twenty-one samples (1.7%) had other psychoactive substances. The analysis of 1179 samples showed that JWH-018 was present in 1172 (99.4%) of the samples. JWH-081 was found in 777 samples (65.9%) together with JWH-018. Samples had different package names. Bonzai Aromatic Potpourri (n = 755; 64.0%) and Bonzai Plant Growth Regulator (n = 316; 26.8%) were the most common product names amongst the herbal products in this study. It is clear from the present study and previous studies that brand name of synthetic cannabinoids that dominate the market exhibit regional differences as to the type and detection frequency of synthetic cannabinoids and the content of herbal highs packages. The number and diversity of synthetic cannabinoid compounds have increased dramatically in the drug market in recent years. New, different, potent derivatives appear on the market, almost every clay and this presents important problems that need to be solved by scientists and judicial authorities working to prevent their harm. These problems include the limited knowledge about their frequency, the lack of analytical data and reference standards for analysis of these new derivates, the lack of information on their toxic effects, and information about the metabolism and metabolites for toxicological analysis in human subjects. (c) 2013 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved

The relationship between age-related development of spike-and-wave discharges and the resistance to amygdaloid kindling in rats with genetic absence epilepsy

Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are resistant to amygdaloid kindling. Since in GAERS the characteristics of spike-and-wave discharges (SWDs) change with age, we have studied the relation between SWD maturation and the development of kindling resistance. Non-epileptic Wistar rats and GAERS were stimulated in basolateral amygdala with 400 mu A at 20 min intervals until they reached stage 5 seizures or for a maximum of 36 stimulations. All of the Wistar rats, the postnatal (PN) day 20 GAERS and the (kindling-prone) subgroups of GAERS at PN30 and PN60 reached stage 5 seizures; at PN20, PN30 and PN60 kindling rates were significantly slower in GAERS compared to Wistar rats. At PN30 and PN60, 41% and 69% of GAERS, respectively, showed no stage 3, 4 or 5 seizures after 36 stimulations (kindling-resistant subgroups). The SWD maturation involves changes in spectral patterns and correlate with age-related increases in kindling resistance in GAERS. (C) 2008 Elsevier Inc. All rights reserved

30 ve 60 günlük yavru Wistar sıçanlarda kindling

AMAÇ: Kindling; düşük şiddette tekrarlayan elektriksel uyarılar ile ortaya çıkan nöbetlerle karakterize, sekonder jeneralize temporal lob epilepsi modelidir.Yetişkin hayvanlarda art arda, sık aralıklarla verilen elektriksel uyarılar (örn:15–30 dakika), kindling gelişimini yavaşlatmakta ya da durdurmaktadır. Yavru sıçanlarda ise sık aralıklarla verilen elektriksel uyarıların kindling gelişimini sağladığı bildirilmektedir. Bu çalışmada kindling gelişim sürecine yaşın ve uyarı aralığının etkisinin gösterilmesi amaçlanmıştır. YÖNTEM VE GEREÇLER: Çalışmada 30 ve 60 günlük erkek Wistar sıçanlar kullanılmıştır. Stereotaksik cerrahi yöntemle hayvanların bazolateral amigdalasına tek taraflı bipolar uyarı ve kayıt, kortekse ise kayıt elektrodu yerleştirilmiştir. İki günlük iyileşme döneminden sonra hayvanlar iki gruba ayrılmıştır; birinci grup 90 dakika, ikinci grup ise 20 dakika aralıklarla 2 saniye süre ile 80 Hz,1 ms,400 mikroamper şiddetinde bir akımla uyarılmıştır. Uyarı öncesi ve sonrasında korteks ve amigdaladan elektroanselografi kaydı alınmıştır. Nöbet evresi Racine’in skalasına göre değerlendirilmiştir. Üç kez 5.evre nöbet geçiren hayvanlar “kindled” kabul edilmişlerdir. BULGULAR: : Tekrarlayan elektriksel uyarılar sonrasında 20 ve 90 dakika aralıklarla uyarılan gruplardaki bütün hayvanlar 5.evre nöbet geçirmiş ve “kindled” olmuşlardır. 20 dakika grubunda sıçanların 5.evre nöbet geçirmesi için gerekli olan ortalama uyarı sayısı 15,3 ± 3, 90 dakika grubunda ise 15,2 ± 1,3 olarak saptanmıştır. Gruplar arasında “kindling” gelişim sürecinde istatistiksel olarak farklılık bulunmamıştır. Sıçanların 5.evre nöbet geçirdiklerinde, amigdaladan kaydedilen art-deşarj süreleri ise 20 dakika grubunda 27.07 ± 3,7 sn, 90 dakika grubunda 34,52 ± 1,5 sn olarak saptanmıştır. Bu fark istatistiksel olarak anlamlı bulunmuştur. SONUÇLAR: : Yavru sıçanlarda 20 ve 90 dakika aralıklarla verilen uyarılar “kindling” oluşturabilmekte, “kindling” evreleme süreci birbirine benzer olmakla beraber art-deşarj süreleri farklılık göstermektedir

Genetik absans epilepsili sıçanlarda kortikal odağa etosüksimid uygulamasının kindling üzerine etkisi

Genetik absans epilepsi modellerinde fokal limbik nöbetlerin sekonder jenerelizasyonuna direnç olduğu ve diken-dalga deşarj aktivitesinin kindling uyarılarından etkilendiği gösterilmiştir. Diken-dalga deşarj aktivitesinin somato-sensoriyal korteksin perioral bölgesinde bir odaktan köken aldığı ve kortikal odağa etosüksimid enjeksiyonları ile bu deşarjların baskılandığı bilinmektedir. Bu çalışmada 30 günlük Strasbourg kökenli genetik absans epilepsili sıçanlarda (GAERS), diken-dalga deşarj odağının etosüksimid ile baskılanmasının, limbik epilepsi modeli olan kindling gelişimine etkisi incelenmiştir. Bu amaçla GAERS’lere stereotaksik cerrahi ile sağ bazolateral amigdalaya uyarı elektrodu, kortikal odağa bilateral enjeksiyon kanülü ve kortekse kayıt elektrodları yerleştirilmiştir. Cerrahi sonrasında bir günlük dinlenme süresinin ardından bazal EEG kaydı alınmış ve kindling deneyleri için artdeşarj eşiği belirlenmiştir. Bu işlemi takiben hayvanların sağ ve sol kortekslerine bilateral etosüksimid enjeksiyonu yapılmış ve daha sonra 20 dk’da bir 400 mikroA şiddetinde olacak şekilde elektriksel olarak uyarılmışlardır. Elektriksel uyarılar sıçanlar Racine’in skalasına göre 5. evre nöbet geçirene kadar tekrarlanmış ya da maksimum 36. uyarıya ulaşılması hedeflenmiştir. Diken-dalga deşarjları ve uyarı sonrası artdeşarj süreleri hesaplanmış, oluşan davranış değişiklikleri değerlendirilmiştir. Etosüksimid uygulanan 30 günlük GAERS’lerde diken-dalga deşarj sayı ve süresi enjeksiyonu takip eden 20. dakikadan itibaren anlamlı derecede baskılanmış ve hayvanların hepsi 5. evre nöbet geçirmişlerdir. Bununla birlikte GAERS grubunda ilk defa 4. ve 5. evre nöbet geçirmek için verilmesi gereken uyarı sayısı kontrol grubuna göre anlamlı derecede fazla bulunmuştur. Bu sonuçlar ile diken-dalga aktivitesinin kaynağı olan kortikal odak baskılandığında, fokal limbik nöbetlerin sekonder jenerelizasyonuna direncin ortadan kalktığı gösterilmiştir. Bu durum absans epilepside limbik epilepsiye direnç mekanizmalarının diken-dalga aktivitesi ile ilişkili olduğunu göstermektedir