Considering this, it is relevant to study which hypothalamic

magnocellular nucleus mediates the cardiovascular mTOR inhibitor response evoked by carbachol microinjection into the BST. Taking that into consideration, we evaluated the hypothesis that PVN and/or SON neurons are part of the neural pathway related to cardiovascular responses following carbachol microinjection into the BST of unanesthetized rats. For this, we investigated cardiovascular responses evoked by carbachol microinjection into the BST before and after PVN or SON pretreatment, either ipsilateral or contralateral in relation to BST microinjection site, with the nonselective neurotransmission blocker cobalt chloride (CoCl2). Microinjection of aCSF into the BST (n = 5) did not affect either MAP (99 ± 2 vs. 98 ± 3 mm Hg, t = 0.2, P > 0.05) or HR (379 ± 11 vs. 352 ± 9 bpm, t = 1.3, P > 0.05) baseline values. However, microinjection of carbachol into the BST caused significant pressor and bradycardiac responses in unanesthetized rats ( Fig. 1). Photomicrography of a coronal brain section showing a representative microinjection site into the BST is presented in Fig. 2. Diagrammatic representation of the BST indicating microinjection sites into the BST of all animals used in the present

study is also shown in Fig. 2. Microinjection of carbachol (n = 6) Veliparib molecular weight into the BST significantly increased plasma vasopressin content (aCSF: 2.3 ± 0.5 pg/mL vs. carbachol: 21.3 ± 3.6 pg/mL, t = 5, P < 0.005), when compared to the control group that received vehicle (aCSF) injection into the BST (n = 6). Microinjection of aCSF into the ipsilateral SON (n = 7) did not affect either MAP (98 ± 2 vs. 101 ± 3 mm Hg, t = 0.5, P > 0.05) or HR

(352 ± 7 vs. 367 ± 11 bpm, t = 1.5, P > 0.05) baseline values. Pretreatment of the ipsilateral SON with aCSF also did not affect the pressor (43 ± 2 vs. 38 ± 2 mm Hg, t = 2.3, P > 0.05) and bradycardiac (− 67 ± 7 vs. − 64 ± 8 bpm, t = 0.2, P > 0.05) response to carbachol microinjection into the BST ( Fig. 1A). Microinjection of CoCl2 into the ipsilateral SON (n = 7) did not affect either MAP (102 ± 2 vs. 100 ± 2 mm Hg, t = 0.6, P > 0.05) or HR (351 ± 6 vs. 356 ± 8 bpm, t = 0.7, P > 0.05) baseline values. However, ipsilateral SON pretreatment with CoCl2 significantly reduced the pressor (44 ± 2 vs. 6 ± 1 mm Hg, t = 16, P < 0.0001) and bradycardiac (− 74 ± 6 vs. − 12 ± 1 bpm, Gemcitabine in vitro t = 10, P < 0.0001) response to carbachol microinjection into the BST ( Fig. 1A). Time-course analysis indicated a significant effect of SON pretreatment with CoCl2 in carbachol cardiovascular effects (ΔMAP: F(1,456) = 468, P < 0.0001 and ΔHR: F(1,456) = 111, P < 0.0001), a significant effect over time (ΔMAP: F(37,456) = 23, P < 0.0001 and ΔHR: F(37,456) = 11, P < 0.0001), and an interaction between treatment and time (ΔMAP: F(37,456) = 20, P < 0.0001 and ΔHR: F(37,456) = 4, P < 0.0001) ( Fig. 1B). Microinjection of aCSF into the contralateral SON (n = 6) did not affect either MAP (100 ± 3 vs.

The fistulotomy was done in the middle of the duodenal papilla roof, 1 cm above the papillar orifice, to gain access to the bile duct. The fistula was enlarged with a standard papillotomy in order to remove the bile duct stones (Figure 1 and Figure 2). ERCP is the standard treatment

for impacted bile duct stones at duodenal papilla. However the impacted PCI-32765 cell line stone can lead to failure of deep cannulation with standard papillotomy and stone extraction. An endoscopic needle-knife fistulotomy can provide an artificial choledocoduodenal fistula thereby facilitating the removal of the stone.2 and 3 It is important after the fistulotomy a complete and large biliary sphincterotomy to permit total stone clearance and to avoid complications. The authors declare that the procedures followed were in accordance with the regulations

of the relevant clinical research ethics committee and with those of the Code of Ethics of the World Medical Association (Declaration of Helsinki). The authors declare that they have followed the protocols of their work center on the publication of patient data and that all the patients included in the study received sufficient information and gave their written informed consent to participate in the study. The authors have obtained the written informed consent of the patients or subjects mentioned in the article. The corresponding author is in possession of this document. The authors have no conflicts of interest to declare. “
“O GE está em mudança. Conforme planeado, a edição passou, desde março de 2012, a ser feita pela prestigiada editora, Elsevier. Parece-nos que conseguimos else assim uma revista de melhor Idelalisib cost qualidade, e também a possibilidade de obter uma forma mais expedita de processar a receção dos artigos, subsequente envio para os revisores, eventual revisão e, finalmente, a publicação. O processo informático de submissão parece inicialmente um pouco complexo, e pedia para isso a vossa compreensão. No entanto, a médio prazo torna-se fácil de utilizar. Procuramos que, desde que o artigo é recebido até à sua publicação,

o tempo não ultrapasse os 4 meses. No momento atual, estamos a recuperar algum atraso, sobretudo no que diz respeito à publicação dos casos clínicos. Temos incentivado a publicação de «guidelines» e normas de atuação em Gastrenterologia por considerarmos ser de grande interesse o seu conhecimento pelos gastrenterologistas. Continuamos a receber um bom número de casos clínicos e de «flashs» endoscópicos. No entanto, gostaríamos de receber mais artigos originais e, nesse sentido, pedimos a vossa colaboração. De facto, tendo como objetivo a indexação da revista, este só será alcançado se aumentarmos a qualidade e o número dos artigos originais. Temos procurado que haja um Editorial por cada artigo publicado, para pôr em perspetiva os achados de investigação publicados, e pensamos que isso tem sido apreciado pelos leitores.

A recent TMS study using intentional

binding as an implicit measure of agency also suggests a contribution of the supplementary motor complex (Moore et al., 2010). But that study was designed Alectinib in vivo to test whether candidate areas were necessary for intentional binding, and could not draw strong anatomical conclusions about the precise location of the neural correlates of implicit agency. Indeed, the repetitive stimulation protocol used in such studies may have rather widespread effects in the stimulated region of cortex (Mochizuki et al., 2005), and can also produce remote effects via neural connections with the stimulated region (Stefan et al., 2008). A recent meta-analysis of studies on the neural correlates of agency as

identified in neuroimaging data has implicated the importance of parietal brain regions such as angular gyrus, TPJ and pre-SMA, but also found an association between agency and activation of the insula, dorsofrontomedian cortex and precuneus (Sperduti et al., 2011). However, this meta-analysis did not focus on low-level implicit markers of sense of agency. We therefore aimed to identify brain regions associated with the implicit sense of agency, taking intentional binding as a proxy for sense of agency. We used an interval estimation task, in which participants judged the time between a button press and a resulting tone. In one condition this tone was elicited by the participant’s active button press, in another condition the tone was U0126 purchase elicited by a passive movement of the same finger (cf. Engbert et al., 2007). In order to extract brain areas associated with the intentional binding effect we used a parametric Phosphatidylinositol diacylglycerol-lyase approach in which we modulated each trial with its respective judgement error. Thus, trials with strong

binding effects would have large and negative values for this regressor, since underestimation of an action–effect interval corresponds to a negative judgement error. The parametric regressor in the passive condition of the interval estimation task is assumed to capture all brain activation responsible for non-specific causes of variation in time estimation, such as arousal, division of attention etc. The parametric regressor for the active condition on the other hand was assumed to identify both these non-specific factors, and additionally the agency-related changes in time perception due to intentional binding. Contrasting these two parametrically modulated conditions – one that shows the attraction of voluntary action and tone, and one that does not – offers the possibility to extract brain regions that are related to intentional binding. We used this technique to investigate the specific contributions of the SMA and the angular gyrus to sense of agency, given that these areas were repeatedly reported in previous studies of agency. Seventeen healthy students (five males; age: mean = 22.

Nevertheless, it is clear that present acquisition and processing methodologies are some way off enabling a reliable quantitative assessment of subtle BBB abnormalities and further work is required to improve these. “
“In the above article, the post-doctoral training Compound Library manufacturer grant number listed in the Acknowledgments section is incorrect. The Acknowledgment should have read: This research was supported in part by a post-doctoral training grant in image science (T32 EB001628) and the Vanderbilt

CTSA (UL1 RR024975-01) NCRR/NIH. “
“In the above article, the second author’s name was misspelled “Siuyan Liu”. It is now printed correctly. The authors regret any inconvenience or confusion this error may have caused. “
“Anxiety and mood disorders contribute substantially to the burden of disease and disability in the United States. A recent national study estimates that generalized anxiety disorder (GAD), posttraumatic stress disorder (PTSD), and major depressive disorder affect 5.7%, 6.8%, and 16.6% of adults in their lifetime, respectively (Kessler et al., 2005). Studies have established a genetic contribution to these mental disorders (Hettema et al., 2001, Sullivan et al., 2000 and Xian et al., 2000). Yet, the mapping of direct paths from

gene to mental disorders has been slow and inconsistent, as only a few genome-wide association studies have detected risk genes and many putative gene findings have failed replication (Hamer, 2002). More fundamentally, a large proportion Venetoclax ic50 of variation in mental health remains unexplained by genetic factors. For these reasons, discovery of new risk factors for mental disorders is crucial. CHIR-99021 cost A growing body of epidemiologic literature has implicated infections as novel risk factors for development of mental disorders (Benros et al., 2013 and Dalman et al., 2008). One pathogen of particular interest is the neurotropic parasite Toxoplasma gondii (T. gondii). T. gondii is capable of reproducing asexually within any warm-blooded animal but must return to its definitive host, the cat, to undergo sexual reproduction,

develop into infectious oocysts, and return to the environment through fecal shedding ( Carruthers and Suzuki, 2007). Infection is transmitted to an intermediate host (e.g., a rodent) or a dead-end host (e.g., a human) via ingestion of tissues cysts in undercooked meat or oocysts in cat feces or contaminated soil, whereupon the parasite progresses to form latent cysts in muscle and neural cells, including neurons, glial cells, and astrocytes ( Carruthers and Suzuki, 2007). As T. gondii does not complete its life cycle until passing from its intermediate rodent host to its definitive feline host, the “manipulation hypothesis” posits that the parasite may be under selective pressure to influence rodent behavior to promote predation by and transmission to the definitive feline host ( Lafferty, 1999). Indeed, T.

These absorbance ratios correspond roughly to the range of CR absorbance ratios (R) encountered in oceanic measurements. The absorbance

Y-27632 cost measurements used to determine the ratios were well within the linear-response characteristics of the Cary 400 spectrophotometer. The temperature and salinity ranges were 278.13 ≤ T ≤ 308.27 K and 20 ≤ S ≤ 40. Initial estimates for the e1 term in Eq.  (2) were obtained by determining the e1 molar absorptivity ratio at a pH where the HI− form of the dye is dominant. Iterative calculations are necessary to account for absorbance contributions at 433 nm and 573 nm from the H2I and I2 − forms of the dye. The overlapping absorbance spectra of H2I, HI− and I2 − are shown in Fig. 1. A speciation model for T = 298.15 K and S = 35 was constructed using the K1 determined as described in Section 2.7 and the K2 reported by Byrne and Breland (1989). At a pH of 4.5, HI− is near

99.91% of the total CR concentration; the fractions of H2I and I2 − are 0.045% and 0.046%. Requisite e1 ABT-199 in vivo absorbance data (573A/433A) were determined with a 0.02 m acetate buffer solution at ionic strength of 0.7 m NaCl. No salinity dependence was observed for the very small e1 term. During preparation of the acetate/acetic acid buffer solution, pHf (free scale) was monitored with a ROSS combination electrode that had been calibrated on the free hydrogen ion scale by titrating a 0.7 m NaCl solution with standard HCl. Because the HI− absorbance signal includes contributions from the H2I and I2 − forms of the dye, the following equation was used to account for these contributions (see also derivation of Liu et al., 2011): equation(6) e1=εHI−573εHI−433=AHI−573/sHI−AHI−433/sHI−=AT573−AH2I573−AI2−573AT433−AH2I433−AI2−433where λεHI is the molar absorptivity at a given wavelength (λ) for the HI− form of the indicator, λAx is the absorbance at wavelength λ of total (T) indicator (all forms) or of individual indicator forms (H2I, HI−, or I2 −), s is the cell pathlength, and [HI−] is the concentration of the HI− form.

Expressing the absorbance terms in Eq. (6) in terms of molar absorptivities and total CR concentrations (IT) via K1 and K2, e1 can be written as follows: isothipendyl equation(7) e1=AT573−εH2I573ITsH+2K1K21+H+K2+H+2K1K2−1−εI2−573ITs1+H+K2+H+2K1K2−1AT433−εH2I433ITsH+2K1K21+H+K2+H+2K1K2−1−εI2−433ITs1+H+K2+H+2K1K2−1 To obtain the K2 value required in this calculation, initial e1 estimates were used to obtain initial K2T estimates by solving Eq.  (2) for − log (K2Te2). The e2 term, required to calculate K2T from − log(K2Te2), was calculated as a function of temperature by using the HI− absorbance at λ = 433 nm in the solution used to determine e1 (i.e., acetate buffer of pH = 4.5 and 0.7 m ionic strength) and the absorbance at λ = 573 nm in the solution used to determine e3/e2 (i.e., modified synthetic seawater of pH = 12 and 0.7 m ionic strength).

If the stenosis affects subclavian artery, changed hemodynamic spectra suggesting subclavian steal syndrome are recorded (Fig. S1 supplementary file). When occlusion of the subclavian artery sets in, in ipsilateral vertebral artery hemodynamic spectra are completely inverse (Fig. S2 supplementary file), and in the contralateral one it is accelerated. Transcranial Doppler of the Willis circle and vertebrobasilar system shows redistribution of the hemodynamics. GCA, is also known as temporal arteritis or cranial arteritis, is the most see more common form of vasculitis that occurs in adults [8]. Almost all patients who develop GCA are over

the age of 50. It is a granulomas arteritis affecting large or medium-sized artery, usually Selleck Galunisertib temporal or ophthalmic artery. It has an acute or subacute start. Symptoms are headache, jaw pain, blurred or double vision. If the disease is undiagnosed complications like blindness and, less often, stroke may occur. Standard test for diagnosing GCA is biopsy of the temporal artery. More samples are needed because the inflammation may not occur in all parts of the artery. Prompt treatment with corticosteroids relieves symptoms and prevents loss of vision. Ultrasound finding will show swelling of the arterial wall presenting as a hypoechoic dark halo around the color coded flow in the temporal, ophthalmic artery or external carotid artery [7] and [9]. The disease is segmental, therefore, its

visualization is suitable for Cell press localization of the biopsy. Due to noninvasiveness it is suitable

for monitoring the disease. During healing regression of the dark halo will be visible parallel with the restitution of the color coded flow. Fibromuscular dysplasia (FMD) is a fibrous thickening of the arterial wall, causing segmental narrowing of arteries in the kidneys (in 75% of patients), carotid or vertebral arteries and the arteries of the abdomen [10]. It is an autosomal dominant disorder, affecting up to 5% of the population, in 2/3 the internal carotid artery (ICA), usually the C2 segment. It is usually asymptomatic, but if dissection occurs, it causes aneurysm and occlusion and becomes symptomatic. There are three types of fibromuscular dysplasia: intimal, medial, and subadventitial (perimedial) of the arterial wall. These three types of FMD are not easily differentiated by findings on angiography. The medial type of FMD is by far the most common (about 80–85%) and it is classically diagnosed on the basis of a “string of beads” appearance on angiography. This appearance is explained by the presence of luminal stenosis alternating with aneurysmal dilatation. Classically, the intimal form of FMD is associated with smooth focal stenoses on angiography. Type 1 is the most common form. In 6–12% of patients with arterial fibroplasia, a long tubular stenosis may be seen. This form is most commonly seen with the intimal form.

Calixto and Siqueira Jr. (2008) have indicated several difficulties in relation to the development of R&D by the Brazilian pharmaceutical industry: high costs and risks associated with the development of new traditional drugs, high financial costs (interest rates) and a low supply of risk capital, the long maturation time of R&D projects, a lack of formal R&D divisions in the industry, a reduction in the number of domestic companies due to mergers with or acquisitions by multinational/transnational corporations, a lack

of experience in technological innovation, the absence of researchers in companies, and a lack of programmes that include the participation of the national government and its agencies. By understanding the role of the Brazilian Ministry of Health in Neglected Diseases R&D, the Department learn more of Science and Technology (DECIT) has supported several projects in this area, through the Secretariat of Science, Technology and Strategic Inputs (SCTIE). Thus, our fibrin sealant has obtained the necessary R&D funding. This scenario was only possible due to the advanced-stage development and translational capacity

of the fibrin selleck screening library sealant and because the Brazilian government is committed to investing in technology and the development of new drugs targeting public health. At the website http://www.clinicaltrials.gov, a total of 119,470 clinical studies were registered between 01/01/1990 Bumetanide and 31/12/2011. Over the same period, Brazil was responsible for only 2720 records on this platform. Regarding

the ability to conduct clinical trials in Brazil, it is observed that only 19.9% of trials were recorded as phase 0, phase I, phase II or phase I + II, while 62.1% of the trials were recorded as phase II + III, phase III or phase IV (ClinicalTrials.gov, 2012). This finding demonstrates that most of the clinical trials conducted in Brazil, representing a small proportion of the studies performed worldwide, involve protocols that reflect the priorities of foreign laboratories. The participation of Brazilian researchers in these studies has been limited to executing protocols developed in other countries. Furthermore, both the analysis and ownership of the data are entirely within the scope of the contracting companies. In this context, there is a great disincentive for the academic community to participate in clinical research. Without financial incentive, physicians often feel undervalued or indifferent to the benefits of performing clinical research for their patients (Kahn et al., 2011). According to Morgan et al. (2011), researchers describe translational research as “high risk” and are seldom viewed by their peers as contributing “authentic” knowledge that would bestow symbolic capital in their field.

, 2008). With respect to smoking as a risk factor, it has long been acknowledged that the use of combustible tobacco products elevates the likelihood of an individual developing cardiovascular disease (Rosamond et al., 2007). This may be linked to exposure to one (or a combination) of a number of cigarette smoke toxicants which modify the activity and function of cells click here within the cardiovascular

system and initiate pathogenic processes. Cigarette smoke is a complex mixture composed of more than 5,600 chemicals (Perfetti and Rodgman, 2011). Within this unique matrix, several chemicals have been identified as toxicants and are thought to drive disease processes (Hoffman and Hecht, 1990). While attempts have been made to identify

those compounds that have the greatest risk of inducing disease, no single toxicant or group of toxicants has been identified as the inducer of cardiovascular disease processes. Specific smoke constituents have been administered to animal models of cardiovascular disease in order to assess their effects on atherosclerotic lesion development (O’Toole et al., 2009 and Srivastava et al., 2011). However, a single compound behaves much differently in a simple state learn more than when it is combined with >5,600 unique compounds with unique properties (e.g., free radicals, antioxidants, toxicants). Moreover, it is further likely that direct interactions with compounds in the complex smoke mixture may have mitigating effects. Since the identity of the compound(s) in cigarette smoke that drive lesion progression remains elusive, an approach that has received considerable attention of late has been the development of potentially reduced-exposure products (PREP). In 2001, the US Institute of Medicine reported that, since smoking-related diseases were dose related, and because epidemiological studies show reduction in the risk of smoking-related diseases following

cessation, it might be possible to reduce smoking-related risks by developing PREPs (Stratton et al., 2001). In this report a framework was proposed for the assessment of the biological effects of cigarettes with modified Thiamet G yields of smoke toxicants. An important component of this approach to product evaluation is the use of in vitro models of smoking-related diseases, including cardiovascular disease. Alongside data from other studies (smoke chemistry evaluation, clinical studies examining biomarkers of both exposure and of biological effect, in vitro and in vivo toxicological studies, in vivo models of disease and epidemiological studies), findings made using in vitro disease models would form part of a weight-of-evidence approach to evaluate and support any proposed change in biological effect. What is lacking from this framework is a detailed insight into not only which models to use but how they would form a part of the overall evaluation framework.

However during acute illness or surgery patients may still be exposed to blood products, although specifically transfusing patients for immunological benefit is no longer routine [18], [19] and [20]. Leucodepletion of blood products has also been shown not to prevent the risk of allosensitisation associated with RBCT [14], [21], [22] and [23]. The majority of studies on the role of blood transfusion was performed in the period before the use of sensitive and specific solid phase antibody detection assays were available and cell-dependent cytotoxicity assays were utilised.

Although it is established that DSA detected at the time of transplant is associated with an increased risk of AMR why some patients with DSA develop AMR and others do not is unclear and may relate to variability in the antibody sub-type, complement SGI-1776 chemical structure binding ability, or the amount or breadth of antibody [1], find more [24], [25] and [26]. Transfusion in the peri-operative and early post-transplant period depends on individualised patient management factors and is commonly thought not to be an immunological stimulus because it is assumed that the concomitant use of immunosuppression mitigates this risk. We hypothesised

that post-transplant transfusion in patients with preformed HLA antibody may provide additional allostimulation or immunological recall and increase the risk of AMR. We therefore investigated the relationship of pre-transplant and peri-operative transfusion in renal transplant

recipients with and without pre-transplant HLA antibody determined by Luminex single antigen bead (SAB) assay. We studied 258 transplant recipients of which 246 patients Resveratrol received a kidney transplant and 12 patients received a simultaneous pancreas–kidney transplant between June 2003 and October 2007. Patients were transplanted at 3 tertiary centres and peri-operative care and decision for transfusions was individualised, clinically indicated and not mandated by protocol. No donor-specific transfusions occurred. Leucocyte depleted packed red cells were used. All patients received a calcineurin inhibitor (CNI) (tacrolimus or cyclosporine) at the time of transplantation in combination with mycophenolate mofetil or mycophenolate sodium and corticosteroids and the Interleukin-2 receptor antibody basiliximab was commonly used for induction. The need for biopsy, medication adjustments and transfusion was determined by the caring clinical teams and was not protocol driven. Transfusion history was obtained from the West Australian Red Cross Blood Bank, the Westmead Hospital Transfusion Laboratory, patient medical records and direct patient interrogation. Patient follow-up was a median of 67 months (IQR 54–77). Patients provided written consent for participation in this study. These are reported in detail elsewhere however stored donor DNA was typed by sequence based typing at HLA-A, -B, -C, -DRB1, DQB1, DPB1 loci and DRB3, 4, 5 and DQA1 where required [27].

1%, triton X-100 0.1% and propidium iodide 50 μg/ml) (Nicoletti et al., 1991), and the cell fluorescence was determined by flow cytometry, as described above. The mitochondrial transmembrane potential was determined by the retention of rhodamine 123 dye (Gorman et al., 1997 and Sureda et al., 1997). The cells were washed this website with PBS, incubated with rhodamine 123 (5 μg/ml, Sigma Chemical Co. St Louis, MO, USA) at 37 °C for 15 min in the dark and washed twice. The cells were then incubated again in PBS at 37 °C for 30 min in the dark and their fluorescence was measured

by flow cytometry, as described above. Phosphatidylserine externalisation was analysed by flow cytometry (Vermes et al., 1995). A Guava® Nexin Assay Kit (Guava Technologies, Hayward, CA) determined www.selleckchem.com/products/PLX-4032.html which cells were apoptotic (early apoptotic + late apoptotic). The cells were washed twice with cold PBS and then re-suspended in 135 μl of PBS with

5 μl of 7-amino-actinomycin D (7-AAD) and 10 μl of Annexin V–PE. The cells were gently vortexed and incubated for 20 min at room temperature (20–25 °C) in the dark. Afterwards, the cells were analysed by flow cytometry, as described above. Caspase 3/7 activity was analysed by flow cytometry using the Guava® EasyCyte Caspase 3/7 Kit (Guava Technologies, Hayward, CA). The cells were incubated with Fluorescent Labelled Inhibitor of Caspases (FLICATM) and maintained for 1 h at 37 °C in a CO2 incubator. After incubation, 80 μl of wash buffer was added and the cells were centrifuged at 2000 rpm for 5 min. The resulting pellet was resuspended in 200 μl of wash buffer and centrifuged. The cells were then re-suspended in the working solution (propidium iodide and wash buffer) and analysed immediately using flow cytometry, as described above. The drop test assay determined the relative sensitivity of different S. cerevisiae strains to ATZD treatment. The following S. cerevisiae strains were used: BY-4741, Top1Δ and Top3Δ. Cells were treated with ATZD Thiamet G at concentrations of 50 and 100 μg/ml and more, 4

dilutions 1:10 were performed. A suspension of 2 × 105 cells/ml of S. cerevisiae in the exponential phase was used. An aliquot of 3 μl of each dilution was added to plates containing YEPD medium (YEL + agar). After 3–4 days of growth at 28 °C, the plates were photographed. m-AMSA served as the positive control. The inhibitory effects of ATZD on human DNA topoisomerase I were measured using a Topo I Drug Screening Kit (TopoGEN, Inc.). Supercoiled (Form I) plasmid DNA (250 ng) was incubated with human Topo I (4 units) at 37 °C for 30 min in relaxation buffer (10 mM Tris buffer pH 7.9, 1 mM EDTA, 0.15 M NaCl, 0.1% BSA, 0.1 mM spermidine and 5% glycerol) in the presence or absence of ATZD (50 and 100 μg/ml, final 20 μl). The concentrations used were based on the positive control indicated in this Kit. CPT (100 μM) served as the positive control.