bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence of human papillomaviruses in urine samples of male patients infected with HIV‐1 in Sao Paulo, Brazil
Authors: Costa Fernando A.M. , da Silva Roberto Carvalho , Arruda Liã B. , Duarte Alberto J.S. , Casseb Jorge
Published: October 2009

Human papillomavirus is a DNA virus that includes 118 genotypes. HPV16 is responsible for 80% of cervical cancer in women. Men are important reservoirs and major transmitters of HPV to their partners. The aim of this study was to detect HPV DNA and to determine the prevalence of HPV types 6, 11, 16, and 18 in urine samples of men infected with HIV‐1. This study included 223 patients infected with HIV‐1 from the Center of Reference on HIV/AIDS (CRT‐SP) and an outpatient clinic of HIV. Urine samples were collected and after DNA extraction real‐time PCR was performed for detection of HPV DNA. Positive samples were then tested by conventional PCR using type‐specific primers for the four HPV types. A total of 223 men infected with HIV‐1 were tested, 81% of whom were on HAART. Four (5.8%) were positive for HPV6, 18 (26.1%) were positive for HPV11, 22 (31.9%) were positive for HPV16 and five (7.2%) were positive for HPV18 by conventional PCR. Twenty (29%) patients had other HPV types and five patients (1.5%) had multiple types. The mean T CD4+cells count was 517 and 441 cells/mm3 (P = 0.30), in HPV negative and positive men, respectively. The HIV viral load was higher in the HPV negative group than for in the men with HPV (P = 0.0002). A 30.9% prevalence of HPV was found in asymptomatic urine samples of men infected with HIV‐1. This study suggests that urine may be a useful specimen for HPV screening. J. Med. Virol. 81:2007–2011, 2009. © 2009 Wiley‐Liss, Inc.
Application of a modified loop‐mediated isothermal amplification kit for detecting Norovirus genogroups I and II
Authors: Yoda Tomoko , Suzuki Yasuhiko , Yamazaki Kenji , Sakon Naomi , Kanki Masashi , Kase Tetsuo , Takahashi Kazuo , Inoue Kiyoshi
Published: October 2009

Norovirus is a major etiologic agent in worldwide outbreaks of gastroenteritis associated with food as well as person‐to‐person transmission. The ubiquitous nature of Norovirus necessitates simple and rapid detection methods with high accuracy and sensitivity. To this end, several investigators have evaluated the usefulness of commercial reverse‐transcription loop‐mediated isothermal amplification (RT‐LAMP) kits for detecting Norovirus genogroups I (GI) and II (GII). In previous studies, the conventional Loopamp kit for Norovirus GII showed a relatively high detection rate, while that for Norovirus GI showed a relatively low detection rate. In the present study, clinical Norovirus specimens were used to compare the detection rate of a modified Loopamp kit for Norovirus GI with the rates of the conventional Loopamp kit for Norovirus GI and an “in‐house” RT‐LAMP GI primer set, methods which had a high detection rate. Results from the present study showed that the modified Loopamp kit for Norovirus GI had a higher detection rate for two viral genotypes (GI.3, GI.11). On comparison with an “in‐house” GII primer set using genotype GII.4 viruses circulating recently, the detection rate by the Loopamp kit for Norovirus GII was found to be higher, with a 98% detection rate. These results indicate the applicability of the modified LAMP kit for GI and the conventional LAMP kit for GII for detection of Noroviruses in clinical samples. J. Med. Virol. 81:2072–2078, 2009. © 2009 Wiley‐Liss, Inc.
Application of an allele‐specific PCR to clinical HIV genotyping samples detects additional K103N mutations in both therapy naïve and experienced patients
Authors: Carr Jillian M. , Green Tara , Shaw David , Daly Lyndal , Hart Wendy , Ratcliff Rodney , Higgins Geoffrey , Burrell Christopher J. , Li Peng , Qiao Ming
Published: October 2009

Low‐level drug resistance is not detected by routine consensus sequence genotype analysis (CSA) but low levels of specific mutations, such as the non‐nucleoside reverse transcriptase inhibitor (NNRTI)‐resistant mutation K103N, can be quantitated by allele‐specific PCR (ASP). This study has applied an ASP to quantitate low‐level K103N in patients presenting for clinical HIV genotyping and assess the correlation with antiretroviral treatment history and outcomes. HIV RNA was extracted from patient plasma and subjected to PCR amplification of the reverse transcriptase (RT) region followed by genotyping by CSA and real‐time ASP for K103N. When applied to samples from patients presenting for genotyping, the ASP detects K103N, not K103 nor K103R, but cross‐reacts with K103S. ASP identified all samples that were K103N by CSA (10.5%) and an additional 14% by ASP only, representing patients who were therapy naïve and with NNRTI treatment history. ASP detected therapy‐acquired K103N at low levels up to 6 years after cessation of NNRTI therapy. In three patients with new HIV diagnosis and K103N detected by ASP only, K103N virus declined rapidly from the circulation but persisted in PBMC DNA at >12 months post‐diagnosis. Efavirenz (EFV) combination therapy in three patients with low‐level K103N suppressed successfully viral load, although one patient developed failure and CSA‐detectable K103N after 15 months of therapy. Thus, analysis of K103N by ASP in conjunction with CSA genotyping provides additional information that reflects K103N transmission and persistence but detection of low‐level K103N does not preclude successful EFV‐containing combination therapy. J. Med. Virol. 81:1983–1990, 2009. © 2009 Wiley‐Liss, Inc.
Rapid changes in rotaviral genotypes in Ecuador
Authors: Hasing Maria Eloisa , Trueba Gabriel , Baquero Maria Ines , Ponce Karina , Cevallos William , Solberg Owen D. , Eisenberg Joseph N.S.
Published: October 2009

Previous studies suggest that the emerging G9P[8] genotype was the most prevalent rotavirus genotype in Ecuador during 2005. This present study provides a temporal analysis of the distribution of rotavirus genotypes in two locations within Ecuador by adding additional years (2006 — early 2008) to the originally reported 2005 data. Data were collected in a rural (northern coastal Ecuador) and urban (Quito) area. In the rural area, a community sample of cases (those presenting diarrhea) and controls (those not presenting diarrhea) were collected between August 2003 and March 2008 resulting in a total of 3,300 stool samples (876 cases and 2,424 controls). Of these samples, 260 were positive for rotavirus by an immunochromatographic test (196 cases and 64 controls). In Quito, 59 fecal samples were collected from children presenting diarrhea and diagnosed with rotavirus. An RT‐PCR analysis of samples collected between 2005 and 2007 suggested that G9 was replaced by G1 and G2 in the rural and urban settings. During this period G9 decreased from 79% to 9% while G2 increased from 0% to 43% in the rural communities, and G9 decreased from 79% to 37% while G2 increased from 3% to 57% in the urban area of Quito. This rapid replacement of G9 by G1 and G2 reinforces the necessity of surveillance to inform vaccination programs. J. Med. Virol. 81:2109–2113, 2009. © 2009 Wiley‐Liss, Inc.
Co‐circulation of three pathogenic hantaviruses: Puumala, Dobrava, and Saaremaa in Hungary
Authors: Plyusnina Angelina , Ferenczi Emöke , Rácz Gábor R. , Nemirov Kirill , Lundkvist Åke , Vaheri Antti , Vapalahti Olli , Plyusnin Alexander
Published: October 2009

Hantaviruses (Bunyaviridae) cause hemorrhagic fever with renal syndrome (HFRS) in Eurasia and hantavirus (cardio)pulmonary syndrome (HCPS) in the Americas. HFRS is caused by Hantaan virus (HTNV), Seoul virus (SEOV), Dobrava virus (DOBV), Saaremaa virus (SAAV), and Puumala virus (PUUV). Of those, only HTNV is not present in Europe. In recent years, hantaviruses, described in other parts of Europe, were also detected at various locations in Hungary. To study the genetic properties of Hungarian hantaviruses in detail, sequences of the viral S and M segments were recovered from bank voles (Myodes glareolus), yellow‐necked mice (Apodemus flavicollis), and striped field mice (Apodemus agrarius) trapped in the Transdanubian region. As expected, the sequences recovered belonged, respectively, to PUUV (two strains), DOBV (one strain), and SAAV (one strain). On phylogenetic trees two new Hungarian PUUV strains located within the well‐ supported Alpe‐Adrian (ALAD) genetic lineage that included also Austrian, Slovenian, and Croatian strains. Analysis of the Hungarian SAAV and DOBV genetic variants showed host‐specific clustering and also geographical clustering within each of these hantavirus species. Hungarian SAAV and DOBV strains were related most closely to strains from Slovenia (Prekmurje region). This study confirms that multiple hantaviruses can co‐circulate in the same locality and can be maintained side‐by‐side in different rodent species. J. Med. Virol. 81:2045–2052, 2009. © 2009 Wiley‐Liss, Inc.
Intrahepatic long‐term persistence of parvovirus B19 and its role in chronic viral hepatitis
Authors: Wang Chun , Heim Albert , Schlaphoff Verena , Suneetha P.V. , Stegmann Kerstin A. , Jiang Hong , Krueger Martin , Fytili Paraskevi , Schulz Thomas , Cornberg Markus , Kandolf Reinhard , Manns Michael P. , Bock C. Thomas , Wedemeyer Heiner
Published: October 2009

Parvovirus B19 (B19V) has been detected in the liver of Asian patients infected with HBV and may contribute to acute and chronic liver disease. This study aimed to investigate the impact of B19V infection in European patients with viral hepatitis. B19V DNA was detected in 1/91 and 0/50 serum samples from patients with chronic hepatitis C and B, respectively. In contrast, B19V DNA was amplified frequently from explanted end‐stage liver tissues (37/50, 74%) and from routine biopsy samples (14/32, 44%) (P < 0.05). However, there was no significant difference in B19V copy number per cell between these two groups. B19V‐specific CD4+ T‐cell responses to two dominant MHC‐class‐restricted epitopes were detected in a similar frequency in healthy anti‐B19V‐positive individuals (3/19; 16%) and patients with chronic hepatitis C (3/13; 23%). These results indicate that B19V can persist in the liver. However, there is no evidence that B19V is a “hepatitis virus” worsening liver disease in European patients with chronic hepatitis C. J. Med. Virol. 81:2079–2088, 2009. © 2009 Wiley‐Liss, Inc.
Prevalence of different HPV types and estimation of prognostic risk factors based on the linear array HPV genotyping test
Authors: Papachristou Eleni , Sypsa Vana , Paraskevis Dimitrios , Gkekas Athanasios , Politi Ekaterini , Nicolaidou Electra , Anifantis Ioannis , Psichogiou Mina , Tsitsika Artemis , Hadjivassiliou Maria , Petrikkos Georgios , Katsambas Andreas , Creatsas Georgios , Hatzakis Angelos
Published: October 2009

The aim of the study was to evaluate the prevalence and risk factors of HPV in a gynecologic population attending outpatient clinics using two new molecular tests. The Amplicor HPV test and the Linear Array (LA) HPV Genotyping test were used for the detection of HPV DNA in 320 women. Multiple logistic regression was used to identify independent prognostic factors of HPV positivity. The agreement between the two methods in terms of their qualitative results was 89.3% (kappa: 0.63). Based on the LA results, the overall prevalence of HPV DNA was 49.1%, 95% confidence interval (95% CI: 43.5%, 54.7%). The prevalence of high‐risk HPV types was 30.3%. The predominant types were HPV‐6 (24.8%) and HPV‐16 (20.4%). Among women with normal cytology, the prevalence of HPV was much higher in those presenting other findings, such as inflammation, than those without other abnormal findings (49.5% vs. 31.5%). On the basis of multivariate analysis, the risk of HPV infection was higher among women with multiple sexual partners [>3 vs. 1: OR = 3.1, 95% CI: (1.5, 7.2)], Pap smear findings [low/high‐grade lesions vs. negative: OR = 2.8, 95% CI: (1.2, 6.5)], the presence of warts [yes vs. no: OR = 3.0, 95% CI: (1.5, 6.3)] and no history of child birth [no vs. yes: OR = 2.6, 95% CI: (1.0, 6.7)]. Younger age was an additional risk factor for HPV infection with carcinogenic genotypes [OR for 1 year increase = 0.93, 95% CI: (0.89, 0.98)]. J. Med. Virol. 81:2059–2065, 2009. © 2009 Wiley‐Liss, Inc.
Evolving clinical landscape of chronic hepatitis B: A multicenter Italian study
Authors: Stroffolini Tommaso , Almasio Piero L. , Sagnelli Evangelista , Mele Alfonso , Gaeta Giovanni Battista ,   and the Italian Hospitals' Collaborating Group
Published: October 2009

The aim of the study was to evaluate the characteristics of chronic hepatitis B with special reference to the geographical origin of the patients and to the prevalence of HBeAg and viral and non‐viral co‐factors of liver disease. A cross‐sectional multicenter survey was undertaken, which enrolled 1,386 HBsAg chronic carriers observed consecutively in 21 referral centers over a 6‐month period. The prevalence of HBeAg in patients was 11%; the presence of HBeAg was associated independently with a younger age and co‐infection with HIV. Anti‐HDV, anti‐HCV, or anti‐HIV antibodies were detected in 8.1%, 6.5%, and 2%, respectively. However, among the patients first diagnosed during the study period (incident cases), 14.3% were anti‐HDV positive. Seven percent of the patients were immigrants; they were younger than Italian patients and 18% were HBeAg positive; no difference was observed in the prevalence of anti‐HDV, anti‐HCV, or anti‐HIV antibodies. The presence of cirrhosis was associated independently with an age >52 years, the presence of anti‐HDV or anti‐HCV, alcohol use >4 drinks/day, and a high BMI. The clinical epidemiology of chronic hepatitis B virus (HBV) infection shows a dynamic profile, with the potential for re‐emergence of cases with HBeAg or anti‐HDV and an emerging impact of metabolic factors on the evolution of liver disease. J. Med. Virol. 81:1999–2006, 2009. © 2009 Wiley‐Liss, Inc.
Monodisperse Hollow Tricolor Pigment Particles for Electronic Paper
Authors: Meng Xianwei , Tang Fangqiong , Peng Bo , Ren Jun
Published: October 2009

A general approach has been designed to blue, green, and red pigments by metal ions doping hollow TiO 2. The reaction involves initial formation of PS at TiO2 core–shell nanoparticles via a mixed-solvent method, and then mixing with metal ions solution containing PEG, followed calcining in the atmosphere. The as-prepared hollow pigments exhibit uniform size, bright color, and tunable density, which are fit for electronic paper display.
Alternating Magnetic Field Controlled, Multifunctional Nano-Reservoirs: Intracellular Uptake and Improved Biocompatibility
Authors: Ghosh Santaneel , GhoshMitra Somesree , Cai Tong , Diercks David R , Mills Nathaniel C , Hynds DiAnna L
Published: October 2009

Biocompatible magnetic nanoparticles hold great therapeutic potential, but conventional particles can be toxic. Here, we report the synthesis and alternating magnetic field dependent actuation of a remotely controllable, multifunctional nano-scale system and its marked biocompatibility with mammalian cells. Monodisperse, magnetic nanospheres based on thermo-sensitive polymer network poly(ethylene glycol) ethyl ether methacrylate-co-poly(ethylene glycol) methyl ether methacrylate were synthesized using free radical polymerization. Synthesized nanospheres have oscillating magnetic field induced thermo-reversible behavior; exhibiting desirable characteristics comparable to the widely used poly-N-isopropylacrylamide-based systems in shrinkage plus a broader volumetric transition range. Remote heating and model drug release were characterized for different field strengths. Nanospheres containing nanoparticles up to an iron concentration of 6 mM were readily taken up by neuron-like PC12 pheochromocytoma cells and had reduced toxicity compared to other surface modified magnetic nanocarriers. Furthermore, nanosphere exposure did not inhibit the extension of cellular processes (neurite outgrowth) even at high iron concentrations (6 mM), indicating minimal negative effects in cellular systems. Excellent intracellular uptake and enhanced biocompatibility coupled with the lack of deleterious effects on neurite outgrowth and prior Food and Drug Administration (FDA) approval of PEG-based carriers suggest increased therapeutic potential of this system for manipulating axon regeneration following nervous system injury.
Plant Stanol Esters Lower Serum Triacylglycerol Concentrations via a Reduced Hepatic VLDL‐1 Production
Authors: Plat Jogchum , Mensink Ronald P.
Published: October 2009

Plant stanol esters not only lower low density lipoprotein cholesterol but also have previously been shown to lower serum triacylglycerol (TAG) concentrations, especially in subjects with elevated TAG concentrations. To find a possible explanation, we explored changes in serum lipoprotein profiles, as measured with nuclear magnetic resonance. For this, serum samples from two parallel‐designed controlled studies were evaluated before and 8 weeks after the consumption of plant stanol esters. In the first study, dyslipidemic metabolic syndrome subjects participated and in the second study normolipidemic subjects. In metabolic syndrome subjects, plant stanol esters lowered concentrations of large (>60 nm) and medium (35–60 nm) VLDL particles as compared to controls. In normolipidemic subjects, the serum concentration of large VLDL‐1 particles was also lowered, although less pronounced. Based on these findings, we hypothesize that the effect of plant stanol esters on serum TAG concentrations origins from a lowered hepatic production of large TAG‐rich VLDL‐1 particles.
Orthopedic management of spina bifida. Part I: Hip, knee, and rotational deformities
Authors: Swaroop Vineeta T. , Dias Luciano
Published: October 2009

Children with spina bifida develop a wide variety of congenital and acquired orthopedic deformities. Among these are hip deformities such as contracture, subluxation, or dislocation. Patients may also have problems with the knee joint, such as knee flexion or extension contracture, knee valgus deformity, or late knee instability and pain. In addition, rotational deformities of the lower extremities, either internal or external torsion, are common as well. This paper will review both the overall orthopedic care of a patient with spina bifida and provide a focused review of the diagnosis and management of the above deformities. In addition, this paper will review the incidence, etiology, classification, and prognosis of spina bifida. The use of gait analysis and orthoses will be covered as well. The forthcoming Part II will cover foot and ankle deformities in spina bifida.
Interindividual variability of drug transporters: Impact on opioid treatment in chronic renal failure
Authors: Ranzani Guglielmina Nadia , De Gregori Simona , De Gregori Manuela , Regazzi Mario , Govoni Stefano
Published: October 2009

Some transmembrane transporters influence the absorption, the tissue distribution and the elimination of opioids, and their genetic variants may be an important factor in therapeutic efficacy and toxicity. Uptake and efflux transporters may facilitate or limit access of opioids to blood—brain barrier (BBB), modulating pain relief. As renal function is crucial in the metabolism and pharmacokinetic profile of any drug, renal failure and end‐stage renal disease may alter drug disposition by affecting protein and tissue binding, and reducing systemic clearance of drugs.
Ultrasound and facet blocks: A review
Authors: Bertini Laura , Baciarello M.
Published: October 2009

Facet or zygapophysial joints are considered to be common sources of chronic spinal pain. In addition to causing localized spinal pain, facet joints may refer pain to adjacent structures. Cervical facet‐joint pain may radiate to the head, neck, and shoulders. Thoracic facets may produce paraspinous mid‐back pain with neuralgic characteristics; and lumbar facet joints may refer pain to the back, buttocks, and proximal lower extremities. Because the facet joint is innervated by the medial branches arising from the posterior rami of the spinal nerve at the same level and a level above the joint, LA blocks of these nerves have been advocated for diagnostic and prognostic purposes. Intra‐articular l—z joint injection with LA has also been proposed as a method for diagnosing facet‐joint pain, with both procedures appearing to provide comparable diagnostic value. Because several blocks are often needed to identify the symptomatic joint or to rule outzygapophysial joint pain, the procedure may expose patients and personnel to considerable radiation doses. In contrast, ultrasound is not associated with exposure to radiation. Ultrasound guidance was recently proposed as a possible alternative to fluoroscopy for the conduct of lumbar medial‐branch block. Ultrasound‐guided techniques require training in specific areas: (1) pattern recognition of the ultrasonographic appearances of anatomic structures, (2) probe handling and scanning skills, and (3) manual dexterity in aligning the needle to the ultrasound beam, thereby enabling the needling of the target. Ultrasound could become an attractive alternative to fluoroscopy or CT scanning, helping to increase the practicability of lumbar facet nerve blocks.
Luteolin, a food‐derived flavonoid, suppresses adipocyte‐dependent activation of macrophages by inhibiting JNK activation
Authors: Ando Chieko , Takahashi Nobuyuki , Hirai Shizuka , Nishimura Kanako , Lin Shan , Uemura Taku , Goto Tsuyoshi , Yu Rina , Nakagami Joji , Murakami Shigeru , Kawada Teruo
Published: October 2009

Interaction between adipocytes and macrophages contributes to the development of insulin resistance in obese adipose tissues. In this study, we examined whether luteolin, food‐derived flavonoid, could suppress the production of inflammatory mediators of the interaction between adipocytes and macrophages. Experiments using a coculture system of adipocytes and macrophages showed that luteolin suppressed the production of inflammatory mediators. In addition, activated macrophages were targets for the suppressive effect of luteolin. Luteolin inhibited the phosphorylation of JNK and suppressed the production of inflammatory mediators in the activated macrophages. The findings indicate that luteolin can inhibit the interaction between adipocytes and macrophages to suppress the production of inflammatory mediators, suggesting that luteolin is a valuable food‐derived compound for the treatment of metabolic syndrome.
Evodiamine inhibits adipogenesis via the EGFR–PKCα–ERK signaling pathway
Authors: Wang Ting , Wang Youxue , Yamashita Hitoshi
Published: October 2009

The molecular mechanism of the anti‐adipogenic effect of evodiamine (which has several capsaicin‐like pharmacological actions) was investigated. The evodiamine effect was not blocked by the specific TRPV1 antagonist capsazepine in 3T3‐L1 preadipocytes, whereas its effect was greatly curtailed by inhibitors of protein kinase C (PKC) and epidermal growth factor receptor (EGFR). Signal analyses showed that evodiamine stimulated the phosphorylation of EGFR, PKCα, and ERK, all of which were reduced by an EGFR inhibitor. Silencing experiments of EGFR mRNA supported the involvement of these signaling molecules in the inhibitory effect of evodiamine. An unidentified mechanism whereby evodiamine inhibits adipogenesis via the EGFR–PKCα–ERK signaling pathway was revealed.
The crystal structure of a hyperthermoactive exopolygalacturonase from Thermotoga maritima reveals a unique tetramer
Authors: Pijning Tjaard , van Pouderoyen Gertie , Kluskens Leon , van der Oost John , Dijkstra Bauke W.
Published: October 2009

The exopolygalacturonase from Thermotoga maritima is the most thermoactive and thermostable pectinase known to date. Here we present its crystal structure at 2.05 Å resolution. High structural homology around the active site allowed us to propose a model for substrate binding, explaining the exo‐cleavage activity and specificity for non‐methylated saturated galacturonate at the non‐reducing end. Furthermore, the structure reveals unique features that contribute to the formation of stable tetramers in solution. Such an oligomerization has not been observed before for polygalacturonases.
Clathrin and AP1B: Key roles in basolateral trafficking through trans‐endosomal routes
Authors: Gonzalez Alfonso , Rodriguez-Boulan Enrique
Published: October 2009

Research following introduction of the MDCK model system to study epithelial polarity (1978) led to an initial paradigm that posited independent roles of the trans Golgi network (TGN) and recycling endosomes (RE) in the generation of, respectively, biosynthetic and recycling routes of plasma membrane (PM) proteins to apical and basolateral PM domains. This model dominated the field for 20 years. However, studies over the past decade and the discovery of the involvement of clathrin and clathrin adaptors in protein trafficking to the basolateral PM has led to a new paradigm. TGN and RE are now believed to cooperate closely in both biosynthetic and recycling trafficking routes. Here, we critically review these recent advances and the questions that remain unanswered.
Formation and regulation of lipid microdomains in cell membranes: Theory, modeling, and speculation
Authors: Fan Jun , Sammalkorpi Maria , Haataja Mikko
Published: October 2009

Compositional lipid microdomains (“lipid rafts”) in plasma membranes are believed to be important components of many cellular processes. The biophysical mechanisms by which cells regulate the size, lifetime, and spatial localization of these domains are rather poorly understood at the moment. Over the years, experimental studies of raft formation have inspired several phenomenological theories and speculations incorporating a wide variety of thermodynamic assumptions regarding lipid–lipid and lipid–protein interactions, and the potential role of active cellular processes on membrane structure. Here we critically review and discuss these theories, models, and speculations, and present our view on future directions.
The retinal structure of channelrhodopsin‐2 assessed by resonance Raman spectroscopy
Authors: Nack Melanie , Radu Ionela , Bamann Christian , Bamberg Ernst , Heberle Joachim
Published: October 2009

Channelrhodopsin‐2 mediates phototaxis in green algae by acting as a light‐gated cation channel. As a result of this property, it is used as a novel optogenetic tool in neurophysiological applications. Structural information is still scant and we present here the first resonance Raman spectra of channelrhodopsin‐2. Spectra of detergent solubilized and lipid‐reconstituted protein were recorded under pre‐resonant conditions to exclusively probe retinal in its electronic ground state. All‐trans retinal was identified to be the favoured configuration of the chromophore but significant contributions of 13‐cis were detected. Pre‐illumination hardly changed the isomeric composition but small amounts of presumably 9‐cis retinal were found in the light‐adapted state. Spectral analysis suggested that the Schiff base proton is strongly hydrogen‐bonded to a nearby water molecule.
The KDEL receptor: New functions for an old protein
Authors: Capitani Mirco , Sallese Michele
Published: October 2009

The KDEL receptor is a seven‐transmembrane‐domain protein that was first described about 20 years ago. Its well‐known function is to retrotransport chaperones from the Golgi complex to the endoplasmic reticulum. Recent studies, however, have suggested that the KDEL receptor has additional functions. Indeed, we have demonstrated that chaperone‐bound KDEL receptor triggers the activation of Src family kinases on the Golgi complex. This activity is essential in the regulation of Golgi‐to‐plasma membrane transport. However, the identification of different KDEL receptor interactors that are inconsistent with these established functions opens the possibility of further receptor activities.
Systems biology of lipid metabolism: From yeast to human
Authors: Nielsen Jens
Published: October 2009

Lipid metabolism is highly relevant as it plays a central role in a number of human diseases. Due to the highly interactive structure of lipid metabolism and its regulation, it is necessary to apply a holistic approach, and systems biology is therefore well suited for integrated analysis of lipid metabolism. In this paper it is demonstrated that the yeast Saccharomyces cerevisiae serves as an excellent model organism for studying the regulation of lipid metabolism in eukaryotes as most of the regulatory structures in this part of the metabolism are conserved between yeast and mammals. Hereby yeast systems biology can assist to improve our understanding of how lipid metabolism is regulated.
A conserved sequence in caveolin‐1 is both necessary and sufficient for caveolin polarity and cell directional migration
Authors: Sun Xing-Hui , Liu Zi-Yang , Chen Hong , Beardsley Andrew R. , Qi Qi , Liu Jun
Published: October 2009

Caveolin‐1 (Cav‐1) plays an important role in the organization of signaling molecules involved in a variety of signaling pathways, including those mediating cell motility. Here we show that amino acids K47–K57 of Cav‐1 are a highly conserved sequence in Cav‐1 and Cav‐3 proteins, and that expression of either K47–K57 deletion Cav‐1 mutant or wild‐type Cav‐2 that lacks this sequence exhibits a non‐polarized distribution pattern. Expression of K47–K57 in Cav‐2 leads to Cav‐2 polarity, suggesting that expression of K47–K57 is sufficient to direct caveolin polarity. Importantly, we show that expression of this sequence is both necessary and sufficient to promote cell directional migration. Thus, our results support the conclusion that Cav‐1 polarity is critical for cell directional migration.
Mechanisms of COPI vesicle formation
Authors: Hsu Victor W. , Yang Jia-Shu
Published: October 2009

Coat Protein I (COPI) is one of the most intensely investigated coat complexes. Numerous studies have contributed to a general understanding of how coat proteins act to initiate intracellular vesicular transport. This review highlights key recent findings that have shaped our current understanding of how COPI vesicles are formed.
Diffusion of a membrane protein, Tat subunit Hcf106, is highly restricted within the chloroplast thylakoid network
Authors: Vladimirou Elina , Li Michael , Aldridge Cassie P. , Frigerio Lorenzo , Kirkilionis Markus , Robinson Colin
Published: October 2009

The thylakoid membrane forms stacked thylakoids interconnected by ‘stromal’ lamellae. Little is known about the mobility of proteins within this system. We studied a stromal lamellae protein, Hcf106, by targeting an Hcf106‐GFP fusion protein to the thylakoids and photobleaching. We find that even small regions fail to recover Hcf106‐GFP fluorescence over periods of up to 3 min after photobleaching. The protein is thus either immobile within the thylakoid membrane, or its diffusion is tightly restricted within distinct regions. Autofluorescence from the photosystem II light‐harvesting complex in the granal stacks likewise fails to recover. Integral membrane proteins within both the stromal and granal membranes are therefore highly constrained, possibly forming ‘microdomains’ that are sharply separated.
Sphingomyelin metabolism at the plasma membrane: Implications for bioactive sphingolipids
Authors: Milhas Delphine , Clarke Christopher J. , Hannun Yusuf A.
Published: October 2009

The plasma membrane (PM) is a major resource for production of bioactive lipids and contains a large proportion of the cellular sphingomyelin (SM) content. Consequently, the regulation of SM levels at the PM by enzymes such as sphingomyelinase (SMase) and SM synthase 2 (SMS2) can have profound effects – both on biophysical properties of the membrane, but also on cellular signaling. Over the past 20 years, there has been considerable research into the physiological and cellular functions associated with regulation of SM levels, notably with regards to the production of ceramide. In this review, we will summarize this research with particular focus on the SMases and SMS2. We will outline what biological functions are associated with SM metabolism/production at the PM, and discuss what we believe are major challenges that need to be addressed in future studies.
Role of p53/FAK association and p53Ser46 phosphorylation in staurosporine‐mediated apoptosis: Wild type versus mutant p53‐R175H
Authors: Fanucchi Stephanie , Veale Robin B.
Published: October 2009

MINT‐7294020: FAK (uniprotkb:Q05397) physically interacts (MI:0915) with p53 (uniprotkb:P04637) by anti‐bait coimmunoprecipitation (MI:0006)
Regulatory role of monoamine neurotransmitters in astrocytic NT‐3 synthesis
Authors: Mele Tina , Čarman‐Kržan Marija , Jurič Damijana Mojca
Published: October 2009

Astrocytes actively control neuronal activity and synaptic transmission and by producing various neurotrophic factors represent an important local cellular source of trophic support in the normal and diseased brain. Our present study showed the ability of astrocytes to synthesize neurotrophin‐3 (NT‐3) and the active involvement of the monoamine neurotransmitters noradrenaline, adrenaline, dopamine, and serotonin, as well as basic intracellular second messenger systems, in the regulation of NT‐3 production in neonatal rat cortical astrocytes.

Using a new NT‐3 specific enzyme‐immunoassay, we showed that neonatal rat cortical and, for comparison, cerebellar astrocytes in primary culture can synthesize NT‐3; the basal cellular content of NT‐3 protein was 23.2 ± 0.4 pg NT‐3/mg cell protein and 23.6 ± 0.9 pg NT‐3/mg cell protein, respectively. The examined neurotransmitters, with the exception of serotonin, were able to potently and transiently increase NT‐3 mRNA and NT‐3 protein content; their maximal effects were dose‐ and time‐dependent. Noradrenaline (1 μM), adrenaline (1 μM), and dopamine (100 μM) showed a maximal increase in NT‐3 cellular content after 6 h treatment causing a 1.9‐, 1.8‐ and 2.7‐fold elevation, respectively. Prior to the observed increase in NT‐3 protein levels, the examined catecholamines increased NT‐3 mRNA levels with maximal effects observed after 1 h (noradrenaline) and 2 h (adrenaline and dopamine) of incubation causing 2.4‐, 2.6‐ and 3‐fold elevation, respectively. Screening different activators of basic intracellular second messenger systems for their influence on NT‐3 synthesis revealed that forskolin (20 μM), dibutyryl cAMP (dBcAMP) (100 μM), as well as calcimycin (1 μM) (Ca2+ ionophore A23187) and phorbol 12‐myristate 13‐acetate (TPA) (100 nM), markedly increased the cellular level of NT‐3 protein. Neurotransmitter‐induced NT‐3 levels were susceptible (to varying degrees) to inhibition by H‐89 (protein kinase A inhibitor) or staurosporin (protein kinase C inhibitor), which led us to conclude that downstream signaling responsible for the stimulation of NT‐3 synthesis by monoamines in astrocytes consists of multiple, complex intracellular mechanisms involving the cAMP/protein kinase A pathway, activation of protein kinase C, as well as mobilization of Ca2+ ions.

Our results indicate for the first time that monoaminergic neurotransmitters play an important role in the regulation of NT‐3 synthesis in cultured rat astrocytes.
Brachycephalic Feline Noses: CT and Anatomical Study of the Relationship between Head Conformation and the Nasolacrimal Drainage System
Authors: Schlueter Claudia , Budras Klaus D , Ludewig Eberhard , Mayrhofer Elisabeth , Koenig Horst E , Walter Alice , Oechtering Gerhard U
Published: October 2009

Aims A study was designed to evaluate the influence of head conformation on the course of the nasolacrimal drainage system (NDS) in 31 brachycephalic and 15 mesocephalic cats using computed tomography (CT), CT-dacryocystography and anatomical methods.

Findings The higher the degree of brachycephalia, the more the facial bones and upper canine teeth are displaced dorsally (ie, the more pronounced the dorsorotation). Dorsorotation leads to abnormal dislocation of the ventral nasal concha and to almost horizontally rotated upper canine teeth, and thus a steeply oriented NDS. In severe brachycephalia the NDS is forced to pass below the canine tooth (adopt a V-shaped course) and the drainage function seems to be inefficient.

Practical relevance The rotation of the upper canine teeth appears to provide a basis for classification of brachycephalia in cats. The authors recommend that breeders avoid breeding from individuals affected by this condition and to give preference to cats with longer facial bones.
Environmental Enrichment: Practical Strategies for Improving Feline Welfare
Authors: Ellis Sarah LH
Published: October 2009

Practical relevance The clinical application of evidence-based enrichment strategies for the domestic cat housed in a variety of confined environments, ranging from the veterinary cage to the domestic home, is of particular importance - both in relation to providing opportunity for appropriate feline behaviour, and in the prevention and treatment of behavioural and associated health problems (eg, feline lower urinary tract disease associated with negative emotional states such as generalised anxiety). Environmental enrichment has gained particular relevance in the light of current animal welfare legislation. For example, in the UK, the Animal Welfare Act 2006 stipulates that owners/keepers have a duty of care to their animal(s) that includes allowing the animal to exhibit normal behaviour patterns.

Evidence base Research into environmental enrichment as a means of improving animal welfare is still very much in its infancy, particularly in relation to the domestic cat. Thus, evidence-based studies are somewhat sparse and more are needed to validate current recommended enrichment practices.

Audience This article aims to assist general veterinary practitioners to recognise how cats respond to confinement, and to understand what constitutes environmental enrichment, to help them implement or advise on appropriate enrichment strategies for cats confined in a hospital cage, home environment (particularly an indoor-only home), or cattery or rescue shelter, based on published evidence to date. [Box: see text]
Treatment Options for Hyperphosphatemia in Feline CKD: What's Out there?
Authors: Kidder Aimee C , Chew Dennis
Published: October 2009

Practical relevance Phosphorus is retained in chronic kidney disease (CKD), promoting renal secondary hyperparathyroidism and eventually resulting in hyperphosphatemia. Most agree that phosphate retention is a major contributor to the progression of CKD in many species and it is well known that hyperphosphatemia is associated with a significant mortality risk in humans with end-stage renal disease.

Patient group Chronic kidney disease is a common ailment of geriatric cats.

Evidence base There is evidence in cats suggesting that the use of a phosphate-restricted diet in IRIS stage 2–3 disease has a beneficial effect on clinical outcome. However, despite the fact that intestinal phosphate binders are commonly used in veterinary practice for patients with CKD, there have been few published reports focusing on the safety and efficacy of these products in veterinary medicine. No phosphorus binders are licensed as medications for dogs or cats. This article draws on data from clinical trials in humans and studies in cats to discuss treatment goals and options for phosphate retention and hyperphosphatemia in feline CKD.

Clinical significance With careful monitoring of serum phosphate and parathyroid hormone, and implementation of phosphate-restricted dietary management and intestinal phosphate binders, progression of CKD and the degree of hyperparathyroidism in cats may be reduced.

Audience Companion animal and feline practitioners are at the forefront in the management of CKD in cats.
The Inappetent Hospitalised Cat: Clinical Approach to Maximising Nutritional Support
Authors: Chan Daniel L
Published: October 2009

Practical relevance Inappetence is one of the most common presenting complaints in clinically ill cats requiring hospitalisation. When prolonged, poor food intake can lead to malnutrition and may be associated with impaired metabolic function, immunosuppression, compromised wound healing, and increased morbidity and mortality. It is important to recognise that inappetence or anorexia is always secondary to another condition, and that treatment goals should be targeted at the primary condition. The current emphasis in the nutritional support of hospitalised inappetent cats is to provide more effective means of increasing nutritional intake -for example, by initiating enteral nutrition via feeding tubes - rather than rely solely on traditional approaches such as increasing palatability of foods or using appetite-stimulating drugs.

Clinical challenges Cats that are ill enough to require hospitalisation are at increased risk of becoming malnourished because of the combined catabolic effects of their disease and poor nutritional intake. This article highlights some of the problems encountered in treating inappetent cats and discusses a clinical approach to providing better nutritional support.

Patient group Inappetence and anorexia are associated with a myriad of clinical conditions in cats and can be seen in individuals of any age or breed.

Equipment Provision of nutritional support to cats may involve the use of feeding tubes such as nasooesophageal or oesophagostomy tubes. In cases where enteral nutrition is not feasible (eg, cats with gastrointestinal failure), parenteral nutrition should be considered.

Evidence base Various studies have documented the high prevalence of inappetence or anorexia in clinically ill cats. Additional studies have linked poor food intake in cats with serious sequelae such as immunosuppression and hepatic lipidosis. More recently, techniques for providing more effective nutritional support, such as oesophagostomy tubes, have been clinically evaluated and shown to be associated with minimal complications.
Direct drug application to the round window: A comparative study of ototoxicity in rats
Authors: Murillo‐Cuesta Silvia , García‐Alcántara Fernando , Vacas Elena , Sistiaga Jon Alexander , Camarero Guadalupe , Varela‐Nieto Isabel , Rivera Teresa
Published: October 2009

Objective

To assess the validity of inducing ototoxicity in rats by applying a sponge soaked in kanamycin and furosemide on the round window.

Study Design

Basic, randomized, nonblind experimental study.

Setting

Animal models of cochlear damage and reliable methods of local drug delivery are fundamental to study hearing loss and to design new therapies.

Subjects and Methods

Four experimental groups of six Wistar rats with different methods of drug administration were used: (1) injection of subcutaneous kanamycin (400 mg/kg) and intravenous furosemide (100 mg/kg); (2) local application of a sponge soaked in saline close to the round window; (3) animals for which the sponge was soaked in a solution containing kanamycin (200 mg/mL) and furosemide (50 mg/mL); and (4) sham‐operated rats. The tympanic bulla was exposed using a ventral approach, and a bullostomy was performed to visualize the round window membrane. Cochlear function was assessed by measuring the auditory brainstem response, and hearing thresholds in response to click and tone burst stimuli were determined as peak and interpeak latencies. At the end of the study, cochlear histology was analyzed.

Results

Systemic administration of kanamycin and furosemide induced profound hearing loss and severe hair cell damage. Local delivery of these ototoxic drugs caused comparable damage but avoided the systemic side effects of the drug. Sham‐operated and saline control animals did not experience functional alterations.

Conclusion

Situating a sponge soaked in kanamycin and furosemide on the round window membrane through the ventral approach is a reliable method to provoke local ototoxicity in rats.
Additional diagnostic value of 18F‐FDG PET‐CT in detecting retropharyngeal nodal metastases
Authors: Chu Hyung Ro , Kim Jin Hwan , Yoon Dae Young , Hwang Hee Sung , Rho Young‐Soo
Published: October 2009

Objective

This study investigated whether preoperative 18fluorine‐fluorodeoxyglucose positron emission tomography‐computed tomography (PET‐CT) scanning improved the diagnosis of retropharyngeal lymph node (RPLN) metastasis in patients with head and neck squamous cell carcinoma (HNSCC) relative to conventional imaging alone.

Study Design

Case series with chart review.

Setting

University hospital cancer center.

Subjects and Methods

Sixty‐three patients with HNSCC underwent RPLN dissection in accordance with our indications and were subsequently divided into two groups: CT/magnetic resonance imaging (MRI) only (group A, n = 33) and CT/MRI with PET‐CT (group B, n = 30). The preoperative radiological findings of each group were compared with the RPLN histopathologic findings, which served as the standard of reference.

Results

RPLN metastasis was confirmed histopathologically in 17 of 63 patients (27.0%): eight from group A and nine from group B. With the additional use of PET‐CT in group B, the sensitivity (88.9%), specificity (85.7%), and accuracy (86.7%) were higher than the respective values in group A (62.5%, 60.0%, and 60.6%). The positive and negative predictive values for group B (72.7% and 94.7%, respectively) were also higher than those for group A (33.3% and 83.3%, respectively). False‐positive results were obtained in 10 patients from group A and three patients from B; false‐negative findings occurred in three patients from group A and one patient from group B. The predictive power of the radiological findings was statistically significant in group B (P = 0.0017), with an odds ratio of 47.987 (95% confidence interval, 4.3‐535.0).

Conclusion

18F‐FDG PET‐CT, when used in combination with CT/MRI, increases diagnostic efficacy in the detection of RPLN metastases and may therefore be useful in screening high‐risk patients.
Hearing outcomes of daily intratympanic dexamethasone alone as a primary treatment modality for ISSHL
Authors: Hong Seok Min , Park Chan Hum , Lee Jun Ho
Published: October 2009

Objective

Intratympanic (IT) dexamethasone offers a good method for avoiding systemic side effects in patients with sudden idiopathic sensorineural hearing loss (ISSHL). However, in most studies, IT was used as a salvage treatment for patients whose hearing failed to improve with the initial systemic treatment, or as an addition to conventional oral steroid protocols. Therefore, we investigated the outcomes of IT steroid administration as a primary treatment modality for ISSHL.

Study Design

Randomized controlled study.

Setting

Tertiary referral center.

Subjects and Methods

Two treatment methods, IT dexamethasone or oral prednisolone, were randomly assigned to 63 participants. A group of 32 patients (IT group) with ISSHL received IT dexamethasone once a day for eight days. A separate group of 31 patients (oral group), received oral prednisolone, also for eight days. We compared the hearing outcomes in the two groups.

Results

We have found that the outcomes for patients treated with IT dexamethasone as a primary treatment modality for the management of ISSHL presented no difference in pure‐tone averages or hearing recovery rate compared with patients treated with oral steroids. However, differences were noted for hearing thresholds according to frequency: the threshold improvement at low frequencies (250, 500, and 1000 Hz) was not statistically significant between groups; at high frequencies, particularly 4000 and 8000 Hz, the threshold improvement was statistically higher in the oral group than in the IT group.

Conclusion

IT dexamethasone might be a good primary treatment method for ISSHL; but, for the management of ISSHL, we should keep in mind the differences in hearing outcomes with regard to frequency.