bluuebunny/crossref_metadata_2025 huggingface dataset:
mRNA deep sequencing reveals 75 new genes and a complex transcriptional landscape in Mimivirus
Authors: Legendre Matthieu , Audic Stéphane , Poirot Olivier , Hingamp Pascal , Seltzer Virginie , Byrne Deborah , Lartigue Audrey , Lescot Magali , Bernadac Alain , Poulain Julie , Abergel Chantal , Claverie Jean-Michel
Published: April 2010

Mimivirus, a virus infecting Acanthamoeba, is the prototype of the Mimiviridae, the latest addition to the nucleocytoplasmic large DNA viruses. The Mimivirus genome encodes close to 1000 proteins, many of them never before encountered in a virus, such as four amino-acyl tRNA synthetases. To explore the physiology of this exceptional virus and identify the genes involved in the building of its characteristic intracytoplasmic “virion factory,” we coupled electron microscopy observations with the massively parallel pyrosequencing of the polyadenylated RNA fractions of Acanthamoeba castellanii cells at various time post-infection. We generated 633,346 reads, of which 322,904 correspond to Mimivirus transcripts. This first application of deep mRNA sequencing (454 Life Sciences [Roche] FLX) to a large DNA virus allowed the precise delineation of the 5′ and 3′ extremities of Mimivirus mRNAs and revealed 75 new transcripts including several noncoding RNAs. Mimivirus genes are expressed across a wide dynamic range, in a finely regulated manner broadly described by three main temporal classes: early, intermediate, and late. This RNA-seq study confirmed the AAAATTGA sequence as an early promoter element, as well as the presence of palindromes at most of the polyadenylation sites. It also revealed a new promoter element correlating with late gene expression, which is also prominent in Sputnik, the recently described Mimivirus “virophage.” These results—validated genome-wide by the hybridization of total RNA extracted from infected Acanthamoeba cells on a tiling array (Agilent)—will constitute the foundation on which to build subsequent functional studies of the Mimivirus/Acanthamoeba system.
DNA methylation of interferon regulatory factors in gastric cancer and noncancerous gastric mucosae
Authors: Yamashita Masaki , Toyota Minoru , Suzuki Hiromu , Nojima Masanori , Yamamoto Eiichiro , Kamimae Seiko , Watanabe Yoshiyuki , Kai Masahiro , Akashi Hirofumi , Maruyama Reo , Sasaki Yasushi , Yamano Hiroo , Sugai Tamotsu , Shinomura Yasuhisa , Imai Kohzoh , Tokino Takashi , Itoh Fumio
Published: April 2010

Interferon regulatory factors (IRFs) are transcription factors known to play key roles in innate and adaptive immune responses, cell growth, apoptosis, and development. Their function in tumorigenesis of gastric cancer remains to be determined, however. In the present study, therefore, we examined epigenetic inactivation of IRF1–9 in a panel of gastric cancer cell lines. We found that expression of IRF4, IRF5, and IRF8 was frequently suppressed in gastric cancer cell lines; that methylation of the three genes correlated with their silencing; and that treating the cells with the demethylating agent 5‐aza‐2′‐deoxycytidine (DAC) restored their expression. Expression of IRF5 in cancer cells was enhanced by the combination of DAC treatment and adenoviral vector‐mediated expression of p53, p63, or p73. Interferon‐γ‐induced expression of IRF8 was also enhanced by DAC. Moreover, treating gastric cancer cells with DAC enhanced the suppressive effects of interferon‐α, interferon‐β, and interferon‐γ on cell growth. Among a cohort of 455 gastric cancer and noncancerous gastric tissue samples, methylation of IRF4 was frequently observed in both gastric cancer specimens and noncancerous specimens of gastric mucosa from patients with multiple gastric cancers, which suggests IRF4 methylation could be a useful molecular marker for diagnosing recurrence of gastric cancers. Our findings indicate that epigenetic IRF inactivation plays a key role in tumorigenesis of gastric cancer, and that inhibition of DNA methylation may restore the antitumor activity of interferons through up‐regulation of IRFs. (Cancer Sci 2010)
Role of β1‐adrenoceptor in increased lipolysis in cancer cachexia
Authors: Cao Dong‐xing , Wu Guo‐hao , Yang Zi‐ang , Zhang Bo , Jiang Yi , Han Yu‐song , He Guo‐dong , Zhuang Qiu‐ lin , Wang Yan‐fu , Huang Zhong‐lin , Xi Qiu‐lei
Published: April 2010

Increased production of hormone‐sensitive lipase (HSL) protein has been demonstrated to be the major cause behind enhanced lipolysis in cancer cachexia. The mechanism governing this alteration is unknown and was presently investigated. This study was conducted to detect the expression of relevant receptors in the adipocytes of cancer cachexia patients, and to elucidate their implication in the increased lipolysis. Gene expressions of β1‐adrenoceptor (ADRB1), β2‐adrenoceptor (ADRB2), β3‐adrenoceptor (ADRB3), α2C‐adrenoceptor (ADRA2C), natriuretic peptide receptor A (NPRA), insulin receptor (INSR), and HSL were determined in adipose tissues of 34 patients by real‐time PCR. Protein levels of ADRB1 and HSL were determined by western blot analysis. β1‐Adrenoceptor (ADRB1) was also detected by immunofluorescence staining. mRNA expressions of both ADRB1 and HSL were approximately 50% elevated selectively in the cachexia group, whereas mRNA levels of the other receptors were unchanged. β1‐Adrenoceptor (ADRB1) protein expression was 1.5‐fold increased in cachexia as compared with the cancer controls, and 3‐fold increased as compared with nonmalignant controls, and was confirmed as a membrane protein in adipocytes by immunofluorescence. Hormone‐sensitive lipase (HSL) protein expression was 2–2.5‐fold increased selectively in cachectic patients. There was a positive correlation between the protein expressions of ADRB1 and HSL. As much as approximately 50% of the variations in HSL protein expression could be explained by variations in ADRB1 protein expression. There was a link between ADRB1 protein level and lipolytic rate. Increased ADRB1 expression may account for some of the functional changes of HSL in patients with cancer cachexia. (Cancer Sci 2010)
Rheumatoid arthritis and birth outcomes: a Danish and Swedish nationwide prevalence study
Authors: Nørgaard M. , Larsson H. , Pedersen L. , Granath F. , Askling J. , Kieler H. , Ekbom A. , Sørensen H. T. , Stephansson O.
Published: April 2010

Abstract. Nørgaard M, Larsson H, Pedersen L, Granath F, Askling J, Kieler H, Ekbom A, Sørensen HT, Stephansson O (Aarhus University Hospital, Denmark, Karolinska Institutet; Karolinska University Hospital, Solna; and Karolinska Institutet, Solna; Stockholm, Sweden). Rheumatoid arthritis and birth outcomes: a Danish and Swedish nationwide prevalence study.J Intern Med2010;268: 329–337.

Objectives. To examine the prevalence of preterm birth, infants with low Apgar score, small for gestational age (SGA) birth, stillbirth and congenital abnormalities in women with rheumatoid arthritis (RA) compared with women without RA.

Design. Prevalence study.

Setting. Combined Sweden and Denmark nationwide from 1994 to 2006.

Subjects. We included 871 579 women with a first‐time singleton birth identified through population‐based healthcare databases.

Main outcome measures. We compared the prevalence of preterm birth, low Apgar score (<7 at 5 min), SGA birth, stillbirth and congenital abnormalities amongst women with RA compared with women without RA using prevalence odds ratio (OR) with 95% confidence interval (95% CI), whilst controlling for maternal age, smoking, parental cohabitation and year. We stratified analyses by period of birth (1994–1997, 1998–2001 and 2002–2006).

Results. Amongst 1199 women with RA, 7.8% gave birth between 32 and 36 gestational weeks (adjusted OR, 1.44; 95% CI, 1.14–1.82), 1.4% gave birth before gestational week 32 (adjusted OR, 1.55; 95% CI, 0.97–2.47), 1.6% had an infant with a low Apgar score (OR, 0.99; 95% CI, 0.95–1.65), 5.9% had an SGA birth (adjusted OR, 1.56; 95% CI, 1.2–2.01), 0.9% experienced stillbirth (adjusted OR, 2.07; 95% CI, 0.98–4.35) and 4.3% gave birth to an infant with congenital abnormalities (adjusted OR,1.32; 95% CI, 0.98–1.79). The OR for congenital abnormalities decreased from 2.57 (95% CI, 1.59–4.16) in 1994–1997 to 1.00 (95% CI, 0.64–1.56) in 2002–2006.

Conclusions. Women with RA had a high prevalence of most adverse birth outcomes. This could be due to inflammatory activity, medical treatment or other factors not controlled for.
The importance of postpartum glucose tolerance testing after pregnancies complicated by gestational diabetes
Authors: McClean S. , Farrar D. , Kelly C. A. , Tuffnell D. J. , Whitelaw D. C.
Published: April 2010

Diabet. Med. 27, 650–654 (2010)

Aims  To review postpartum glucose tolerance in women with gestational diabetes and evaluate the role of formal 75 g oral glucose tolerance testing vs. fasting plasma glucose in screening for persistent abnormalities.

Methods  Retrospective study of 985 pregnancies over a 10 year period in a mixed ethnic cohort of women who underwent follow‐up glucose tolerance testing at 6 weeks postpartum. Diagnosis obtained by oral glucose tolerance test was tested against that from the fasting plasma glucose value.

Results  There were 272 abnormal postpartum oral glucose tolerance test results (27.6%), with 109 women identified as having frank diabetes. Eleven of these (10%) had fasting plasma glucose ≤6.0 mmol/l, as did 62 of 114 cases of impaired glucose tolerance. A fasting plasma glucose concentration of ≥6.1 mmol/l correctly identified abnormal glucose tolerance in 199 of 272 cases (sensitivity 0.73). South Asian women were much more likely to have persistent abnormalities of glucose tolerance than were Europeans (32 vs. 15%, χ2P < 0.0001).

Conclusions  A postpartum fasting plasma glucose measurement alone is not sensitive enough in our population to classify glucose tolerance status accurately. A formal postpartum oral glucose tolerance test is therefore needed to facilitate early detection and treatment.
Diabetes prevalence, diabetes regimen and co‐morbidity in depressed patients compared with non‐depressed patients in primary care in the Netherlands
Authors: Adriaanse M. C. , Bosmans J. E.
Published: April 2010

Diabet. Med. 27, 718–722 (2010)

Aims  To assess the prevalence of diabetes in depressed patients compared with non‐depressed matched controls and to compare diabetes regimen and co‐morbidity in depressed patients with diabetes vs. non‐depressed patients with diabetes in primary care in the Netherlands.

Methods  A retrospective case–control study over 3 years (2002–2004). Data for depressed patients (n = 7128) and non‐depressed matched controls (n = 23772) were available from an electronic medical record system of 20 general practices organized in one large primary care organization in the Netherlands. Matching was based on year and month of birth, sex and general practitioner. Diabetes, diabetes regimen (i.e. oral glucose medication, insulin or both) and co‐morbidity were defined using Anatomical Therapeutic Chemical classification codes of delivered medication.

Results  The prevalence of diabetes was 5.5% (n = 393) among depressed patients, which was 2.6 times higher than in non‐depressed matched controls where the figure was 2.1% (n = 494; P < 0.001). Diabetes regimen, i.e. the proportions of subjects on oral glucose medication, insulin or both did not differ among depressed patients with diabetes (51, 27 and 22%, respectively) compared with non‐depressed patients with diabetes (51, 30, 19%; P = 0.53). Co‐morbidity was significantly more prevalent among depressed patient with diabetes than in non‐depressed patients with diabetes.

Conclusions  Compared with non‐depressed matched controls, adults with treated depression have almost three times higher rates of diabetes. Depressed patients with diabetes had more co‐morbidities compared with non‐depressed patients with diabetes, whereas diabetes regimen did not differ.
Novel monogenic diabetes mutations in the P2 promoter of the HNF4A gene are associated with impaired function in vitro
Authors: Wirsing A. , Johnstone K. A. , Harries L. W. , Ellard S. , Ryffel G. U. , Stanik J. , Gasperikova D. , Klimes I. , Murphy R.
Published: April 2010

Diabet. Med. 27, 631–635 (2010)

Aims  Mutations in HNF4A cause a form of monogenic β‐cell diabetes. We aimed to identify mutations in the pancreas‐specific P2 promoter of HNF4A in families with suspected HNF4A diabetes and to show that they impaired the function of the promoter in vitro.

Methods  We screened families with a clinical suspicion of HNF4A monogenic β‐cell diabetes for mutations in the HNF4A P2 promoter. We investigated the function of the previously reported HNF4A P2 promoter mutation −192C>G linked to late‐onset diabetes in several families, along with two new segregating mutations, in vitro using a modified luciferase reporter assay system with enhanced sensitivity.

Results  We identified two novel HNF4A P2 promoter mutations that co‐segregate with diabetes in two families, −136A>G and −169C>T. Both families displayed phenotypes typical of HNF4A monogenic β‐cell diabetes, including at least two affected generations, good response to sulphonylurea treatment and increased birthweight and/or neonatal hypoglycaemia. We show that both of these novel mutations and −192C>G impair the function of the promoter in transient transfection assays.

Conclusions  Two novel mutations identified here and the previously identified late‐onset diabetes mutation, −192C>G, impair the function of the HNF4A P2 promoter in vitro.
Estimating the direct cost of Type 2 diabetes in Greece: the effects of blood glucose regulation on patient cost
Authors: Athanasakis K. , Ollandezos M. , Angeli A. , Gregoriou A. , Geitona M. , Kyriopoulos J.
Published: April 2010

Diabet. Med. 27, 679–684 (2010)

Aims  To estimate the annual cost of treatment for Type 2 diabetic patients in Greece and investigate the effect of blood glucose regulation on patient cost.

Methods  A multipoint data collection procedure based on the patient records of 51 geographically distributed physicians was used in order to obtain the necessary data for the analysis and the construction of the patient cost model. Patients were classified as controlled (i.e. maintaining blood glucose regulation for the 1 year retrospective time frame of the analysis) and non‐controlled (the patients failing to do so in the specified time period). Cost categories included pharmaceutical expenditure, laboratory/diagnostic tests and consultation fees. Costs attributable to hospitalizations due to diabetic complications were not included. Calculations were based on 2007 fees and prices, and costs are expressed in Euros.

Results  The average annual cost of treatment for controlled patients was estimated at 981.72€ (95% confidence interval, 940.66–1023.01€), whereas for non‐controlled patients it was 1566.12€ (95% confidence interval, 1485.42–1650.20€). Non‐controlled patients had 29.7% higher annual pharmaceutical costs (340.50 vs. 441.96€), 70% higher costs for laboratory/diagnostic tests (422.54 vs. 718.49€) and 85.5% higher consultation costs (218.68 vs. 405.67€) compared with their controlled peers. The average cost for a Type 2 diabetic patient in Greece, regardless of blood glucose regulation, was 1297.30€ (95% confidence interval, 1244.42–1349.61€).

Conclusions  Failing to control blood glucose levels within ‘glycaemic goals’, apart from the clinical consequences, can also have a significant financial impact, resulting in a 59.5% increase in the mean annual patient cost.
Is risk of diabetic retinopathy lower in Australia’s Greek‐born migrants?
Authors: Brazionis L. , Rowley K. , Itsiopoulos C. , O’Dea K.
Published: April 2010

Diabet. Med. 27, 660–665 (2010)

Aim  The standardized cardiovascular disease death rate for the Greek population in Crete has increased since the 1960s, unlike the all‐cause and cardiovascular disease death rate for Australia’s Greek migrant population, which has remained paradoxically low. A small window of opportunity remains in which the vascular profile of this interesting atypical migrant population can be characterized. This study assessed whether ethnicity modulates the risk of diabetic retinopathy in Greek‐born migrants to Australia.

Methods  The study design was a community‐based cross‐sectional study of diabetic retinopathy in 107 Greek‐born and Australian‐born men with Type 2 diabetes, aged 44–83 years. Diabetic retinopathy was assessed by mydriatic three‐field retinal photography.

Results  Prevalence of diabetic retinopathy was lower in Greek‐born than in Australian‐born participants (22 and 37%, respectively). Despite having a higher mean systolic blood pressure level (148 vs. 137 mmHg), Greek‐born men had a significantly lower risk of diabetic retinopathy than Australian‐born men, after adjusting for age, duration of diabetes, glycated haemoglobin, systolic blood pressure, diastolic blood pressure, albumin to creatinine ratio, and total cholesterol and triglyceride levels [odds ratio 0.32 (0.10–0.99); r2 = 0.41, P = 0.047].

Conclusion  Greek ethnicity may confer some protection against diabetic retinopathy to Australia’s Greek‐born migrants, an effect not explained by established risk factors for diabetic retinopathy. A small window of opportunity remains in which to elucidate the ethnicity‐related exposures that modulate vascular risk in this older migrant population.
Frequency of biochemical hypoglycaemia in adults with Type 1 diabetes with and without impaired awareness of hypoglycaemia: no identifiable differences using continuous glucose monitoring
Authors: Choudhary P. , Geddes J. , Freeman J. V. , Emery C. J. , Heller S. R. , Frier B. M.
Published: April 2010

Diabet. Med. 27, 666–672 (2010)

Objective  Impaired awareness of hypoglycaemia (IAH) is a major risk factor for severe hypoglycaemia in Type 1 diabetes. Although biochemical hypoglycaemia is asserted to be more frequent in IAH, this has not been estimated accurately. The aim of this study was to use Continuous Glucose Monitoring (CGM) to quantify hypoglycaemia in IAH and evaluate its use in identifying impaired awareness of hypoglycaemia.

Methods  Ninety‐five patients with Type 1 diabetes were classified as having normal (n = 74) or impaired awareness (n = 21) using an established method of assessing hypoglycaemia awareness. Hypoglycaemia exposure was assessed prospectively over 9–12 months using weekly 4‐point capillary home blood glucose monitoring (HBGM), 5 days of CGM and prospective reporting of severe hypoglycaemia. The frequencies of biochemical and severe hypoglycaemia were compared in patients with normal and impaired awareness of hypoglycaemia.

Results  Patients with impaired awareness had a 3‐fold higher incidence of severe hypoglycaemia than those with normal awareness [incidence rate ratio (IRR) 3.37 (95% CI 1.30–8.7); P = 0.01] and 1.6‐fold higher incidence of hypoglycaemia on weekly HBGM [IRR 1.63 (95% CI 1.09–2.44); P = 0.02]. No significant differences were observed with CGM [IRR for sensor glucose ≤ 3.0 mmol/l 1.47 (95% CI 0.91–2.39); P = 0.12; IRR for sensor glucose ≤ 2.2 mmol/l 1.23 (95% CI 0.76–1.98); P = 0.40].

Conclusions  Patients with Type 1 diabetes with impaired awareness had a 3‐fold higher risk of severe hypoglycaemia and 1.6‐fold higher incidence of biochemical hypoglycaemia measured with weekly glucose monitoring compared with normal awareness, but 5 days of CGM did not differentiate those with impaired from those with normal awareness.
Type 2 diabetes prevalence and incidence among adults in Taiwan during 1999–2004: a national health insurance data set study
Authors: Chang C. H. , Shau W. Y. , Jiang Y. D. , Li H. Y. , Chang T. J. , H.‐H. Sheu W. , Kwok C. F. , Ho L. T. , Chuang L. M.
Published: April 2010

Diabet. Med. 27, 636–643 (2010)

Aim  To evaluate annual prevalence and incidence of Type 2 diabetes and to examine possible trends among adults in Taiwan.

Methods  A retrospective nationwide longitudinal study using the Taiwan National Health Insurance Research Database collected during 1999–2004. Adult patients aged ≥ 20 years old with prevalent and incident Type 2 diabetes were identified using ICD‐9‐CM diagnostic codes. Age‐specific and age‐direct‐standardized annual incidence and prevalence were calculated to describe their trends in different gender and age group and compared using Poisson regression.

Results  During the study years, the age‐standardized prevalence of Type 2 diabetes increased from 4.7 to 6.5% for men and from 5.3 to 6.6% for women. The increasing trends in prevalence were significant and higher among people aged < 40 and ≥ 80 years. The age‐standardized incidence rates of Type 2 diabetes per 1000 person‐years were approximately 7.6 and remain stable for men, but decreasing from 7.7 to 6.9 for women. However, the incidence increased significantly in younger adults aged < 40 years whose relative incidence (RI with 95% confidence interval) was 1.31 (1.20–1.42) for men and 1.04 (1.01–1.08) for women. The incidence trends for people aged ≥ 40 years were decreased for men and women. The differences in incidence trends between age groups and between genders were all statistically significant (all P < 0.001).

Conclusions  This study demonstrated a substantial increasing trend in Type 2 diabetes prevalence during 1999–2004 among adults in Taiwan. Despite the incidence decreased in older people, young men aged 20–40 years were most susceptible to higher incidence of Type 2 diabetes.
Ectopic expression of α‐synuclein affects the migration of neural stem cells in mouse subventricular zone
Authors: Tani Momo , Hayakawa Hideki , Yasuda Toru , Nihira Tomoko , Hattori Nobutaka , Mizuno Yoshikuni , Mochizuki Hideki
Published: April 2010

J. Neurochem. (2010) 115, 854–863.

α‐Synuclein (α‐syn) is a key protein in Parkinson’s disease (PD), and its abnormal accumulation is implicated only not in the loss of dopaminergic neurons in the substantia nigra but also in impairment of olfactory bulb (OB) in PD. Olfactory dysfunction could arise from these OB changes as an early symptom in PD. We reported previously the impairment of neuronal stem cell (NSC) proliferation in the subventricular zone, which is upstream of OB in PD models. Reduction of NSC generation could potentially lead to olfactory dysfunction, which is commonly associated with and precedes the motor symptoms by several years in PD. Here, we investigated neurosphere formation in vitro and migration of NSCs in vivo after transduction of α‐syn‐encoding retroviral vector to characterize the function of α‐syn in NSC. Over‐expression of α‐syn caused less effective formation of neurospheres and induced morphological changes. Fluorescence‐activated cell sorting showed diminished NSC cell cycle progression induced by over‐expression of α‐syn. Intriguingly, suppression of NSC migration along the rostral migratory stream was observed when the α‐syn‐encoding vector was directly injected into the subventricular zone of mice in vivo. These results indicate that α‐syn affects the generation of NSC and suggest that this protein could serve as a tool for the design of potentially useful therapy for PD patients.
Ketones prevent synaptic dysfunction induced by mitochondrial respiratory complex inhibitors
Authors: Kim Do Young , Vallejo Johana , Rho Jong M.
Published: April 2010

J. Neurochem. (2010) 114, 130–141.

Ketones have previously shown beneficial effects in models of neurodegenerative disorders, particularly against associated mitochondrial dysfunction and cognitive impairment. However, evidence of a synaptic protective effect of ketones remains lacking. We tested the effects of ketones on synaptic impairment induced by mitochondrial respiratory complex (MRC) inhibitors using electrophysiological, reactive oxygen species (ROS) imaging and biochemical techniques. MRC inhibitors dose‐dependently suppressed both population spike (PS) and field potential amplitudes in the CA1 hippocampus. Pre‐treatment with ketones strongly prevented changes in the PS, whereas partial protection was seen in the field potential. Rotenone (Rot; 100 nmol/L), a MRC I inhibitor, suppressed synaptic function without altering ROS levels and PS depression by Rot was unaffected by antioxidants. In contrast, antioxidant‐induced PS recovery against the MRC II inhibitor 3‐nitropropionic acid (3‐NP; 1 mmol/L) was similar to the synaptic protective effects of ketones. Ketones also suppressed ROS generation induced by 3‐NP. Finally, ketones reversed the decreases in ATP levels caused by Rot and 3‐NP. In summary, our data demonstrate that ketones can preserve synaptic function in CA1 hippocampus induced by MRC dysfunction, likely through an antioxidant action and enhanced ATP generation.
P2X1 receptor mobility and trafficking; regulation by receptor insertion and activation
Authors: Lalo Ulyana , Allsopp Rebecca C. , Mahaut‐Smith Martyn P. , Evans Richard J.
Published: April 2010

J. Neurochem. (2010) 113, 1177–1187.

P2X1 receptors for ATP contribute to signalling in a variety of cell types and following stimulation undergo rapid desensitisation (within 1 s), and require ∼5 min to recover. In HEK293 cells P2X1 receptors C‐terminally tagged with enhanced green fluorescent protein (P2X1‐eGFP) were predominantly expressed at the cell surface. Following > 90% photo‐bleaching of P2X1‐eGFP within a 6 μm2 circle at the cell surface fluorescence recovery after photo‐bleaching (FRAP) was fit with a time constant of ∼60 s and recovered to ∼75% of pre‐bleach levels. Following activation of the P2X1 receptor with α,β‐methylene ATP the associated calcium influx doubled the FRAP recovery rate. The protein synthesis inhibitor cycloheximide had only a small effect on repeated FRAP and indicated a limited contribution of new P2X1 receptors to the FRAP. Inhibition of trafficking with brefeldin A reduced recovery and this effect could be reversed following receptor activation. In contrast, the dynamin inhibitor dynasore had no effect on FRAP under unstimulated conditions but reduced the level of recovery following agonist stimulation. In functional studies both brefeldin A and dynasore increased the recovery time from desensitisation. Taken together these studies demonstrate for the first time an important role of receptor recycling on P2X1 receptor responsiveness.
Enhancing the GLP‐1 receptor signaling pathway leads to proliferation and neuroprotection in human neuroblastoma cells
Authors: Li Yazhou , Tweedie David , Mattson Mark P. , Holloway Harold W. , Greig Nigel H.
Published: April 2010

J. Neurochem. (2010) 113, 1621–1631.

Increasing evidence suggests that glucagon‐like peptide‐1 (GLP‐1), an incretin hormone of current interest in type 2 diabetes, is neuroprotective in both cell culture and animal models. To characterize the neuroprotective properties of GLP‐1 and associated underlying mechanisms, we over‐expressed the GLP‐1 receptor (GLP‐1R) on human neuroblastoma SH‐SY5Y cells to generate a neuronal culture system featuring enhanced GLP‐1R signaling. In GLP‐1R over‐expressing SH‐SY5Y (SH‐hGLP‐1R#9) cells, GLP‐1 and the long‐acting agonist exendin‐4 stimulated cell proliferation and increased cell viability by 2‐fold at 24 h at physiologically relevant concentrations. This GLP‐1R‐dependent action was mediated via the protein kinase A and phosphoinositide 3‐kinase signaling pathways, with the MAPK pathway playing a minor role. GLP‐1 and exendin‐4 pretreatment dose‐dependently protected SH‐hGLP‐1R#9 cells from hydrogen peroxide (H2O2)‐ and 6‐hydroxydopamine‐induced cell death. This involved amelioration of elevated caspase 3 activity, down‐regulation of pro‐apoptotic Bax and up‐regulation of anti‐apoptotic Bcl‐2 protein. In the presence of 6‐hydroxydopamine, GLP‐1’s ability to lower caspse‐3 activity was abolished with the phosphoinositide 3‐kinase inhibitor, LY2940002, and partly reduced with the protein kinase A inhibitor, H89. Hence, GLP‐1R mediated neurotrophic and anti‐apoptotic actions co‐contribute to the neuroprotective property of GLP‐1 in neuronal cell cultures, and reinforce the potential therapeutic value of GLP‐1R agonists in neurodegenerative disorders involving oxidative stress.
Cannabis Use Correlates of Syringe Sharing among Injection Drug Users
Authors: Jutras‐Aswad Didier , Zang Geng , Bruneau Julie
Published: April 2010

This study examines whether the relation between acute cannabis use and syringe sharing during single injection days is similar among regular and nonregular users, participating in a cohort study of injection drug users in Montréal, Canada. 236 (36.6%) subjects were classified as regular cannabis users (RCUs), 227 (35.2%) as nonregular cannabis users (NRCUs), and 181 (28.1%) were abstinent. Cannabis use during a single injection day was associated with a fivefold increased risk of sharing (OR 4.92; 1.83–13.22) in NRCUs compared to RCUs. Our results indicate that cannabis use history should be considered when evaluating its potential effect on risk‐taking behaviors. (Am J Addict 2010;19:231–237)
Co‐occurring Patterns of Smoking and Alcohol Consumption among Korean Adolescents
Authors: Chung Ick‐Joong , Chun JongSerl
Published: April 2010

This study seeks to assess the transition probabilities between smoking and alcohol consumption trajectories for ages 13–17 using data from the Korea Youth Panel Survey (KYPS). Four smoking trajectories were identified—noninitiator, late‐onsetter, experimenter, and escalator. Similarly, four alcohol consumption trajectories were identified—noninitiator, late‐onsetter, experimenter, and chronic user. Those in the chronic group of alcohol consumption were most likely to be smokers. Those who fell into a particular group for use of one substance were most likely to fall into the corresponding group for use of the other substance. Implications for smoking and alcohol prevention are discussed. (Am J Addict 2010;19:252–256)
Prevalence and Correlates of Withdrawal‐Related Insomnia among Adults with Alcohol Dependence: Results from a National Survey
Authors: Brower Kirk J. , Perron Brian E.
Published: April 2010

Insomnia during acute alcohol withdrawal (AWD) as well as persisting insomnia during postacute withdrawal is associated with relapse. Rates of insomnia in clinical samples of alcohol‐dependent patients range from 36% to 91%, but the prevalence of AWD‐related insomnia in the general population is unknown. The purpose of this study was to describe the prevalence of insomnia as a symptom of acute AWD and its correlates in a general population of alcohol‐dependent individuals. Data were analyzed from the 2001 to 2002 National Epidemiologic Survey on Alcohol and Related Conditions, which sampled 43,093 adults. The prevalence of AWD‐related insomnia among individuals with a lifetime diagnosis of alcohol dependence was 31.7%, which ranked fourth among the eight listed DSM‐IV withdrawal symptoms. Among individuals who met lifetime criteria for both alcohol dependence and AWD, the prevalence of insomnia was approximately 50%. Lifetime diagnoses of major depression and drug use disorders were significant correlates of AWD‐related insomnia in multivariate analyses. A less than 1‐year duration of the heaviest drinking period as well as the onset of alcohol dependence between ages 18 and 27 were negatively associated with AWD‐related insomnia. AWD‐related insomnia is a common symptom among alcohol‐dependent adults in the general population and is related to lifetime co‐occurring diagnoses, age at onset of alcohol dependence, and duration of heaviest drinking period. Its prevalence in the general population provides a representative base rate against which to compare the widely varying rates reported in clinical populations. Because of its relatively frequent prevalence and association with relapse, assessment and treatment of AWD‐related insomnia should be routinely considered in clinical settings. (Am J Addict 2010;19:238–244)
OCDS Craving Scores Predict 24‐Month Outcome in Alcoholic Outpatients
Authors: Soyka Michael , Helten Claudia , Schmidt Peggy
Published: April 2010

This study was conducted to replicate previous findings on the predictive value of a German version of the Obsessive Compulsive Drinking Scale (OCDS) by investigating 24‐month treatment outcome in an outpatient setting. This was a prospective, observational study with 92 alcohol‐dependent patients. The OCDS was used to assess craving at the end of treatment, and at the 6‐, 12‐, and 24‐month follow‐ups. Univariate and logistic regression analyses were performed. Of the 67 patients interviewed at the 24‐month follow‐up, 58% were abstinent and 79% improved. OCDS scores were higher in patients with a less favorable outcome. In line with previous findings, our results showed that the intensity of craving as measured by the OCDS may predict outcome in outpatient alcoholics. (Am J Addict 2010;19:264–269)
Survey of Eating Disorder Symptoms among Women in Treatment for Substance Abuse
Authors: Cohen Lisa R. , Greenfield Shelly F. , Gordon Susan , Killeen Therese , Jiang Huiping , Zhang Yulei , Hien Denise
Published: April 2010

A strong association between substance use disorders (SUDs) and eating disorders (EDs) in women has been established. Yet, little is known about the rates and impact of ED symptoms in women presenting to addiction treatment. The current investigation assessed the prevalence of ED symptoms and their effect on treatment outcomes in a sample of substance abusing women with co‐occurring posttraumatic stress disorder (PTSD) enrolled in outpatient substance use programs. Participants were 122 women who participated in a multisite clinical trial comparing two behavioral treatments for co‐occurring SUD and PTSD. The Eating Disorder Examination‐self report, and measures of PTSD and SUD symptoms were administered at baseline, during treatment and at four follow‐up points. Two subgroups emerged; those reporting binge eating in the 28 days prior to baseline (Binge group; n = 35) and those who reported no binge eating episodes (No Binge group; n = 87). Women in the Binge group endorsed significantly higher ED, PTSD, and depression symptoms at baseline than those in the No Binge group. Although all participants showed significant reductions in PTSD symptoms and improvements in abstinence rates during the study period, the improvements for the Binge group were significantly lower. These findings suggest that a subgroup of women with co‐occurring PTSD and SUDs, who endorsed binge ED symptoms, responded differently to SUD/PTSD group treatment. Identification of ED symptoms among treatment‐seeking women with SUDs may be an important element in tailoring interventions and enhancing treatment outcomes. (Am J Addict 2010;19:245–251)
Exploring the Mediational Role of Coping Motives for Marijuana Use in Terms of the Relation between Anxiety Sensitivity and Marijuana Dependence
Authors: Johnson Kirsten , Mullin Jennifer L. , Marshall Erin C. , Bonn‐Miller Marcel O. , Zvolensky Michael
Published: April 2010

This study evaluated the prediction that coping motives for marijuana use would mediate the relation between anxiety sensitivity and a marijuana dependence diagnosis after controlling for other co‐occurring marijuana use motives. Participants were 136 current marijuana users (47.1% women; Mage= 21.9, SD = 7.2). Results were consistent with a mediational effect, with the relation between anxiety sensitivity and marijuana dependence being explained by the addition of coping motives into the model. These results provide novel information related to the putative explanatory role of coping motives for marijuana use in the relation between anxiety sensitivity and marijuana dependence. (Am J Addict 2010;19:277–282)
Susceptibility of brainstem to kindling and transfer to the forebrain
Authors: Lam Ann , Whelan Naomi , Corcoran Michael E.
Published: April 2010

Purpose:  The kindling of seizures with stimulation of brainstem sites has been reported inconsistently in the literature. The characteristics of the kindling observed, involving high intensities of stimulation and immediate onset of generalized tonic–clonic convulsions, raise questions regarding the nature of kindling from these sites.

Methods:  We implanted chronic electrodes in either the nucleus reticularis pontis oralis (RPO), mesencephalic reticular formation (MRF), dorsal periaqueductal gray (dPAG), or ventrolateral periaqueductal gray (vlPAG) in male Long‐Evans rats, with a recording electrode in the amygdala. Rats received conventional high‐frequency kindling stimulation once daily for 30 days. To test for transfer, we kindled the amygdala beginning 7 weeks after the last brainstem kindling trial.

Results:  Tonic–clonic seizures were evoked by stimulation from all brainstem sites. Seizures were brief and were associated with characteristic low‐amplitude high‐frequency afterdischarge (AD). Kindling of the dPAG resulted in the development of classic AD and increased AD duration. Prior kindling of the dPAG facilitated subsequent kindling of the amygdala; however, no transfer was observed with prekindling of other brainstem sites.

Discussion:  The variability in the response to kindling stimulation suggests that certain brainstem sites are resistant to kindling, whereas other sites are more susceptible to kindling but are still relatively resistant in comparison to sites in the forebrain. The development of classic AD in later trials of dPAG stimulation suggests that epileptogenesis can occur even in the initial absence of classic AD when low‐amplitude high‐frequency AD is present.
The topography and significance of extratemporal hypometabolism in refractory mesial temporal lobe epilepsy examined by FDG‐PET
Authors: Wong Chong H. , Bleasel Andrew , Wen Lingfeng , Eberl Stefan , Byth Karen , Fulham Michael , Somerville Ernest , Mohamed Armin
Published: April 2010

Purpose:  This study aims to map the temporal and extratemporal 18‐fluorodeoxyglucose positron emission tomography (FDG‐PET)–defined hypometabolism in mesial temporal lobe epilepsy (MTLE). We hypothesize that quantitative analysis will reveal extensive extratemporal glucose hypometabolism (EH), that the EH is related to seizure propagation beyond the temporal lobe, hypometabolism restricted to one temporal lobe predicts a good outcome following surgery, and EH predicts a poor outcome.

Methods:  Sixty‐four patients were studied who had undergone temporal lobectomy for intractable MTLE and had at least 2 years of postoperative follow‐up. Spatial preprocessing and statistical analysis on preoperative interictal FDG‐PET using statistical parametric mapping (SPM 2) identified significant regions of hypometabolism compared to normal controls. The predictors of outcome were determined by univariable and multiple logistic regression analyses.

Results:  EH was common and widespread, occurring most frequently in the ipsilateral insula and frontal lobe. The extent of EH was not significantly associated with age of onset or the duration of epilepsy. Presence of secondarily generalized tonic‐–clonic seizures (SGTCS) was associated with a larger extent of remote hypometabolism (RH, p < 0.005). Multiple logistic regression analysis identified the extent of RH and the age at surgery as independent predictors of seizure outcome.

Discussion:  Our results indicate that RH in MTLE is associated with a poorer surgical outcome, especially if seen in the contralateral hemisphere. The extent of RH relates to SGTCS but not to duration of epilepsy.
When is a postoperative seizure equivalent to “epilepsy recurrence” after epilepsy surgery?
Authors: Jehi Lara , Sarkis Rani , Bingaman William , Kotagal Prakash , Najm Imad
Published: April 2010

Purpose:  Up to one‐half of epilepsy surgery patients will have at least one seizure after surgery. We aim to characterize the prognosis following a first postoperative seizure, and provide criteria allowing early identification of recurrent refractory epilepsy.

Methods:  Analyzing 915 epilepsy surgery patients operated on between 1990 and 2007, we studied 276 who had ≥1 seizure beyond the immediate postoperative period. The probability of subsequent seizures was calculated using survival analysis. Patients were divided into seizure‐free (no seizures for ≥1 year) and refractory (persistent seizures) and analyzed using multivariate regression analysis.

Results:  After a first seizure, 50% had a recurrence within 1 month and 77% within a year before the risk slowed down to additional 2–3% increments every two subsequent years. After a second seizure, 50% had a recurrence within 2 weeks, 78% within 2 months, and 83% within 6 months. Having both the first and second seizures within six postoperative months [odds ratio (OR) 4.04; 95% confidence interval (CI) 2.05–8.40; p = 0.0001], an unprovoked initial recurrence (OR 3.92; 95% CI 2.13–7.30; p < 0.0001), and ipsilateral spikes on a 6‐months postoperative electroencephalography (EEG) (OR 2.05; 95% CI 1.10–3.88; p = 0.025) predicted a poorer outcome, with 95% of patients who had all three risk factors becoming refractory. All patients with cryptogenic epilepsy and recurrent seizures developed refractoriness.

Discussion:  Seizures will recur in most patients who present with their first postoperative event, with one‐third eventually regaining seizure‐freedom. Etiology and early and unprovoked postoperative seizures with epileptiform activity on EEG at six postoperative months may predict recurrent medical refractoriness.
Can changes in cortical excitability distinguish progressive from juvenile myoclonic epilepsy?
Authors: Badawy Radwa A.B. , Macdonell Richard A.L. , Jackson Graeme D , Berkovic Samuel F.
Published: April 2010

Purpose:  We used transcranial magnetic stimulation (TMS) to investigate whether there were any characteristic cortical excitability changes in progressive myoclonic epilepsy (PME) compared to juvenile myoclonic epilepsy (JME).

Methods:  Six patients with PME were studied. Motor threshold (MT) at rest and recovery curve analysis using paired‐pulse stimulation at a number of interstimulus intervals (ISIs) was determined. Results were compared to those of 9 patients with chronic refractory JME and 10 with chronic well‐controlled JME.

Results:  PME showed a marked increase in cortical excitability at all the long ISIs (p < 0.01), compared to refractory JME (effect sizes ranging from 1.4 to 1.9) and well‐controlled JME (effect sizes ranging from 2.0 to 2.4). Significant differences at the short ISIs 2–5 ms were seen only on comparison with the well‐controlled group (p < 0.05, effect size 0.6, 0.7). There were no significant differences in MTs of PME compared to either JME groups.

Conclusion:  Our findings demonstrate specific differences in cortical excitability using TMS between PME and those with JME, particularly at long latencies in the paired‐pulse paradigm, implicating a role for γ‐aminobutyric acid (GABA)B‐mediated networks.
Frameshift mutations of the ARX gene in familial Ohtahara syndrome
Authors: Kato Mitushiro , Koyama Norihisa , Ohta Masayasu , Miura Kiyokuni , Hayasaka Kiyoshi
Published: April 2010

Purpose:  Ohtahara syndrome is one of the most severe and earliest forms of epilepsy and is frequently associated with brain malformations, such as hemimegalencephaly. Recently, longer expansion of the first polyalanine tract of ARX was found to be causative for Ohtahara syndrome without brain malformation, whereas premature termination mutations of ARX were found to cause severe brain malformations, such as lissencephaly or hydranencephaly. Both are designated as ARX‐related interneuronopathies.

Methods:  We investigated the molecular basis of Ohtahara syndrome in two families, comprising six male patients in two generations demonstrating X‐linked inheritance.

Results:  Novel frameshift mutations in the terminal exon of the ARX gene (Ala524fsX534 and E536fsX672) were identified in two patients (2 and 13 years, each) from both families. Two patients developed West syndrome, and one of these later developed Lennox‐Gastaut syndrome. Brain magnetic resonance imaging (MRI) of all patients showed no brain malformations in contrast to the patients with a premature termination mutation in other exons of ARX.

Discussion:  The etiology of Ohtahara syndrome is heterogeneous; however, the molecular analysis of ARX should be considered in sporadic or familial male patients with Ohtahara syndrome.
Alteration of NMDA receptor–mediated synaptic interactions in the lateral amygdala associated with seizure activity in a mouse model of chronic temporal lobe epilepsy
Authors: Graebenitz Stéphanie , Lesting Jörg , Sosulina Ludmila , Seidenbecher Thomas , Pape Hans‐Christian
Published: April 2010

Purpose:  Because results from both animal models and human temporal lobe epilepsy (TLE) have pointed to synaptic network alterations in the amygdala, we have tested the hypothesis that glutamatergic transmission in the lateral amygdala (LA) is critically involved.

Methods:  Using the pilocarpine mouse model, LA slices were prepared ex vivo in the recurrent phase of TLE (Pilo group), and LA projection neurons (PNs) were recorded using patch‐clamp techniques. Intrinsic and synaptic properties of LA PNs were analyzed and compared with those in age‐matched saline‐injected controls.

Results:  Only mild changes were observed in intrinsic properties of LA PNs, whereas both spontaneous excitatory postsynaptic currents (sEPSCs) and miniature EPSCs (mEPSCs) were significantly increased in Pilo as compared to saline controls. This difference was sensitive to AP5, but persisted during action of NBQX, indicating mediation by N‐methyl‐d‐aspartate (NMDA) receptors. Moreover, these changes were associated with an increase in frequency but not amplitude of mEPSCs, indicative of a contribution of presynaptic mechanisms.

Discussion:  In conclusion, dynamic changes seem to occur in glutamatergic transmission within the amygdala during TLE, to which a functional upregulation of presynaptic NMDA receptors in LA PNs makes a significant contribution.
Brain development in children with new onset epilepsy: A prospective controlled cohort investigation
Authors: Hermann Bruce P. , Dabbs Kevin , Becker Tara , Jones Jana E. , Myers y Gutierrez Adan , Wendt Gary , Koehn Monica A. , Sheth Raj , Seidenberg Michael
Published: April 2010

Purpose:  To characterize prospective neurodevelopmental changes in brain structure in children with new and recent‐onset epilepsy compared to healthy controls.

Methods:  Thirty‐four healthy controls (mean age 12.9 years) and 38 children with new/recent‐onset idiopathic epilepsy (mean age 12.9 years) underwent 1.5 T magnetic resonance imaging (MRI) at baseline and 2 years later. Prospective changes in total cerebral and lobar gray and white matter volumes were compared within and between groups.

Results:  Prospective changes in gray matter volume were comparable for the epilepsy and control groups, with significant (p < 0.0001) reduction in total cerebral gray matter, due primarily to significant (p < 0.001) reductions in frontal and parietal gray matter. Prospective white matter volume changes differed between groups. Controls exhibited a significant (p = 0.0012) increase in total cerebral white matter volume due to significant (p < 0.001) volume increases in the frontal, parietal, and temporal lobes. In contrast, the epilepsy group exhibited nonsignificant white matter volume change in the total cerebrum (p = 0.51) as well as across all lobes (all p’s > 0.06). The group by white matter volume change interactions were significant for total cerebrum (p = 0.04) and frontal lobe (p = 0.04).

Discussion:  Children with new and recent‐onset epilepsy exhibit an altered pattern of brain development characterized by delayed age‐appropriate increase in white matter volume. These findings may affect cognitive development through reduced brain connectivity and may also be related to the impairments in executive function commonly reported in this population.