bluuebunny/crossref_metadata_2025_split huggingface dataset:
Production of Foamy Virus Vector and Transduction of Hematopoietic Cells
Authors: Josephson Neil C. , Russell David W. 
Published: September 2010

INTRODUCTIONFoamy viruses (FVs), or spumaviruses, are nonpathogenic retroviruses that have been developed as integrating viral vectors. This protocol presents methods for producing high-titer FV vector stocks, free of contaminating replication-competent retrovirus, to be used for transducing hematopoietic stem cells. FV vector stocks are produced by transfecting 293 cells, harvesting and filtering the culture medium, and concentrating vector virions by ultracentrifugation. The resulting stocks are free of replication-competent helper virus, as indicated by a sensitive marker rescue assay. A typical stock made from 23 10-cm dishes has a final volume of 2 mL with a titer of 107 to 108 transducing units/mL. Potential advantages of FV vectors include a lack of pathogenicity of the wild-type virus, a wide host range, stable virions that can be concentrated by centrifugation, a double-stranded DNA genome that is reverse-transcribed in the vector-producing cells, and the largest packaging capacity of any retrovirus. FV vectors are especially useful for transducing hematopoietic cells. Because hematopoietic stem cells have the ability to self-renew, proliferate, and repopulate the bone marrow after transplantation, efficient transduction of these cells offers the promise to cure many inherited and acquired diseases.
Solubilization of Escherichia coli Recombinant Proteins from Inclusion Bodies
Authors: Simpson Richard J. 
Published: September 2010

INTRODUCTIONThe high levels of expression produced by molecular cloning techniques make bacteria particularly useful for producing recombinant proteins. However, these proteins often are difficult to purify, owing to their tendency to aggregate and precipitate within the bacteria as insoluble inclusion bodies. Formation of inclusion bodies is especially common for nonbacterial proteins. Although no single method can be applied to every protein, a number of strategies are available to solubilize inclusion body proteins. This protocol describes one such method.
Focal Recording of Synaptic Currents from Single Boutons at the Drosophila Neuromuscular Junction
Authors: Zhang Bing , Stewart Bryan 
Published: September 2010

INTRODUCTIONThe Drosophila larval neuromuscular junction (NMJ) shares many structural and functional similarities to synapses in other animals, including humans. These include the basic feature of synaptic transmission, as well as the molecular mechanisms regulating the synaptic vesicle cycle. Because of its large size, easy accessibility, and the well-characterized genetics, the fly NMJ remains an excellent model system for dissecting the cellular and molecular mechanisms of synaptic transmission. Focal recording is an extracellular method designed for the study of synaptic activity of one or a few synaptic boutons rather than the ensemble activity of all of the boutons, as occurs with intracellular recording or the two-electrode voltage-clamp (TEVC) method. This is a useful technique for investigating the properties of different motor neurons that innervate the same muscle, applying statistical analysis to discrete synaptic events, and investigating the heterogeneity of synaptic release properties among boutons.
In Vivo Imaging of Cerebral Hemodynamics Using High-Frequency Micro-Ultrasound
Authors: Sun John , Lindvere Liis , van Raaij Martijn E. , Dorr Adrienne , Stefanovic Bojana , Foster F. Stuart 
Published: September 2010

INTRODUCTIONAssessment of cerebral vascular response is important in neuroscience research. Some imaging modalities that are commonly used to detect flow and/or vessel diameter changes in the brain include magnetic resonance imaging, positron emission tomography, and optical intrinsic signal imaging. Ultrasound has not typically been used to assess neurovascular response but recent advances in the technology have led to the development of micro-ultrasound systems with significant potential for this application. The state of the art in high frequency (15-50 MHz) micro-ultrasound is based on linear arrays specifically designed for small animal imaging. These systems can achieve axial resolution ranging from 30 to 200 μm. They are capable of quantifying brain hemodynamics in terms of red blood cell (RBC) velocity, flow, and vascular density in real time, up to 35 mm below the cortical surface, and can achieve temporal resolution of up to 1000 frames per second. This protocol describes imaging of the rat brain using various ultrasound imaging modes (power Doppler, color Doppler, pulsed-wave Doppler, and nonlinear contrast-enhanced imaging) to assess the state of cerebral microcirculation.
Immediate versus delayed quitting and rates of relapse among smokers treated successfully with varenicline, bupropion SR or placebo
Authors: Gonzales David , Jorenby Douglas E. , Brandon Thomas H. , Arteaga Carmen , Lee Theodore C. 
Published: September 2010

ABSTRACTAims  We assessed to what degree smokers who fail to quit on the target quit date (TQD) or lapse following TQD eventually achieve success with continued treatment.Design  A secondary analysis of pooled data of successful quitters treated with varenicline (306 of 696), bupropion (199 of 671) and placebo (121 of 685) from two identically‐designed clinical trials of varenicline versus bupropion sustained‐release and placebo.Setting  Multiple research centers in the US.Participants  Adult smokers (n = 2052) randomized to 12 weeks drug treatment plus 40 weeks follow‐up.Measurement  The primary end‐point for the trials was continuous abstinence for weeks 9–12. TQD was day 8. Two patterns of successful quitting were identified. Immediate quitters (IQs) were continuously abstinent for weeks 2–12. Delayed quitters (DQs) smoked during 1 or more weeks for weeks 2–8.Findings  Cumulative continuous abstinence (IQs + DQs) increased for all treatments during weeks 3–8. Overall IQs and DQs for varenicline were (24%; 20%) versus bupropion (18.0%, P = 0.007; 11.6%, P < 0.001) or placebo (10.2%, P < 0.001; 7.5%, P < 0.001). However, DQs as a proportion of successful quitters was similar for all treatments (varenicline 45%; bupropion 39%; placebo 42%) and accounted for approximately one‐third of those remaining continuously abstinent for weeks 9–52. No gender differences were observed by quit pattern. Post‐treatment relapse was similar across groups.Conclusions  Our data support continuing cessation treatments without interruption for smokers motivated to remain in the quitting process despite lack of success early in the treatment.
Smoothelin is a specific and robust marker for distinction of muscularis propria and muscularis mucosae in the gastrointestinal tract
Authors: Montani Matteo , Thiesler Thore , Kristiansen Glen 
Published: September 2010

Montani M, Thiesler T & Kristiansen G (2010) Histopathology 57, 244–249 Smoothelin is a specific and robust marker for distinction of muscularis propria and muscularis mucosae in the gastrointestinal tract Aims:  As tumour specimens and biopsy specimens become smaller, recognition of anatomical structures relevant for staging is increasingly challenging. So far no marker is known that reliably discriminates between muscularis propria (MP) and muscularis mucosae (MM) of the gastrointestinal tract. Recently, smoothelin expression has been shown to differ in MP and MM of the urinary bladder. We aimed to analyse the expression of smoothelin in the gastrointestinal tract in MP and MM in order to define a novel diagnostic tool to identify MM bundles.Methods and results:  The expression of smoothelin was analysed immunohistochemically in comparison with α‐smooth muscle actin (α‐SMA) in specimens from colon, stomach and oesophagus (n = 107). In contrast to α‐SMA, which equally stained MM and MP, absent or significantly weaker smoothelin expression was found in MM was found, which was particularly valid in colon and gastric specimens.Conclusions:  The combination of smoothelin and SMA represents a robust marker to discriminate MM from MP in the gastrointestinal tract.
Histological grading of breast cancer on needle core biopsy: the role of immunohistochemical assessment of proliferation
Authors: Kwok T’ng Chang , Rakha Emad A , Lee Andrew H S , Grainge Matthew , Green Andrew R , Ellis Ian O , Powe Desmond G 
Published: September 2010

Kwok T C, Rakha E A, Lee A H S, Grainge M, Green A R, Ellis I O & Powe D G (2010) Histopathology57, 212–219 Histological grading of breast cancer on needle core biopsy: the role of immunohistochemical assessment of proliferation Aims:  Histological grade assessed on needle core biopsy (NCB) moderately concurs with the grade in the surgical excision specimen (SES) (κ‐values between 0.35 and 0.65). A major cause of the discrepancy is underestimation of mitoses in the NCB specimen. The aim was to determine the best method of assessing proliferation on NCB.Methods and results:  Proliferative activity of 101 invasive carcinomas of the breast on NCB and SES was assessed using mitotic counts on routine haematoxylin and eosin (H&E) sections and immunohistochemical markers Mib‐1 and phosphorylated histone H3 (PPH3). H&E mitotic count in SES was considered as the gold standard. H&E mitotic count was found to be underestimated on NCB when compared with that in SES (P < 0.001), but no significant difference was detected between NCB and SES regarding Mib‐1 (P = 0.13) or PPH3 (P = 0.073). Using receiver–operating characteristic curve, Mib‐1 on NCB was found to agree with the gold standard significantly better than routine H&E on NCB.Conclusions:  Immunohistochemical markers in NCB showed better concordance with H&E mitotic count in SES (gold standard) than routine H&E mitotic count in NCB. Further refinement of cut‐offs and scoring methods is needed.
Expression of Bcl‐2 family proteins and association with clinicopathological characteristics of oral squamous cell carcinoma
Authors: Coutinho‐Camillo Cláudia Malheiros , Lourenço Silvia Vanessa , Nishimoto Ines Nobuko , Kowalski Luiz Paulo , Soares Fernando Augusto 
Published: September 2010

Coutinho‐Camillo C M, Lourenço S V, Nishimoto I N, Kowalski L P & Soares F A (2010) Histopathology57, 304–316 Expression of Bcl‐2 family proteins and association with clinicopathological characteristics of oral squamous cell carcinoma Aims:  To characterize the expression of proteins that inhibit (Bcl‐2, Bcl‐x, Bcl‐xL, Bcl‐2‐related protein A1, BAG‐1) or promote (Bak, Bax, Bim/Bod, Bim‐Long, Bad, Bid, PUMA) apoptosis and determine possible correlations between the expression of these proteins and clinicopathological features of oral squamous cell carcinoma (OSCC).Methods and results:  Two‐hundred and twenty‐nine cases of OSCC, arranged in a tissue microarray, were immunohistochemically analysed. The results demonstrated that the absence of vascular invasion was associated with increased expression of Bak, Bax, Bcl‐xL, Bcl‐2‐related protein and PUMA. Increased expression of Bim/Bod and BAG‐1 was associated with the presence of perineural infiltration. An increase in Bid and Bim‐Long expression was associated with moderately to well‐differentiated tumours. Increased expression of the Bcl‐2‐related protein and PUMA was associated with tumours occurring in the floor of mouth and increased expression of PUMA was also associated with recurrence of the tumour. Multivariate Cox analysis demonstrated that PUMA and Bim‐Long were independent factors in prognosis of OSCC.Conclusions:  Our results showed the involvement of the Bcl‐2 family of proteins in OSCC tumorigenesis and suggest that the expression of apoptotic molecules might be used as a prognostic indicator for OSCC.
Specificity of DOG1 (K9 clone) and protein kinase C theta (clone 27) as immunohistochemical markers of gastrointestinal stromal tumour
Authors: Wong Newton A C S , Shelley‐Fraser Golda 
Published: September 2010

Wong N A C S & Shelley‐Fraser G (2010) Histopathology 57, 250–258 Specificity of DOG1 (K9 clone) and protein kinase C theta (clone 27) as immunohistochemical markers of gastrointestinal stromal tumour Aims:  DOG1 and protein kinase C (PKC) theta are both sensitive immunohistochemical markers of gastrointestinal stromal tumour (GIST). However, there are conflicting data regarding the specificity of the most commonly used PKC theta antibody (clone 27), and there are no existing data regarding the specificity of the only known commercially available DOG1 antibody (K9 clone) at the time of writing. This study’s aim was to characterize the immunoreactivity patterns of both monoclonal antibodies amongst a wide range of neoplasm types including, in particular, histological mimics of GIST.Methods and results:  Immunohistochemistry for DOG1 and PKC theta was performed on whole tissue sections from 23 different neoplasm types (total of 125 cases). Ten of these neoplasm types showed CD117 immunopositivity. Only three (Ewing’s sarcoma, glomus tumour and synovial sarcoma) of the 23 neoplasm types showed DOG1 immunopositivity, and such positivity was often focal and weak in intensity. In contrast, all but four (ganglioneuromas, leiomyomas, desmoplastic small round cell tumours and PEComa/angiomyolipomas) of the 23 neoplasm types showed PKC theta immunopositivity.Conclusions:  Compared with CD117, DOG1 (using the K9 antibody) is a more specific marker, whereas PKC theta (using the clone 27 antibody) is a considerably less specific immunohistochemical marker for GIST.
DOG1 and CD117 are the antibodies of choice in the diagnosis of gastrointestinal stromal tumours
Authors: Novelli Marco , Rossi Sabrina , Rodriguez‐Justo Manuel , Taniere Philippe , Seddon Beatrice , Toffolatti Luisa , Sartor Chiara , Hogendoorn Pancras C W , Sciot Raf , Van Glabbeke Martine , Verweij Jaap , Blay Jean Yves , Hohenberger Peter , Flanagan Adrienne , Dei Tos Angelo P 
Published: September 2010

Novelli M, Rossi S, Rodriguez‐Justo M, Taniere P, Seddon B, Toffolatti L, Sartor C, Hogendoorn P C W, Sciot R, Van Glabbeke M, Verweij J, Blay J Y, Hohenberger P, Flanagan A & Dei Tos A P (2010) Histopathology57, 259–270 DOG1 and CD117 are the antibodies of choice in the diagnosis of gastrointestinal stromal tumoursAims:  The histopathological diagnosis of gastrointestinal stromal tumours (GIST) is typically made based on a combination of clinical and morphological features supported by immunohistochemistry studies. The aim of this study was to examine the staining quality, sensitivity, specificity and utility of antibodies used commonly in GIST diagnosis.Methods and results:  Immunohistochemistry with a panel of antibodies [CD117, DOG1, protein kinase C (PKC)‐theta, nestin, CD34, smooth muscle actin (SMA), desmin, S100 and CD171] was performed on whole sections from 187 GIST and 29 gastrointestinal mesenchymal tumours, and on several microarrays including 355 GISTs and 120 soft tissue sarcomas. Results showed that DOG1 and CD117 were the most sensitive and specific antibodies used in GIST diagnosis. PKC‐theta and nestin were sensitive, but less specific, also staining other spindle cell tumours commonly considered in the differential diagnosis of GIST. CD34 staining was less sensitive than many of the other antibodies and of limited aid in diagnosis. The smooth muscle markers SMA and desmin, together with the neural marker S100, were unhelpful in confirming a diagnosis of GIST, but were particularly useful in the exclusion/diagnosis of other gastrointestinal mesenchymal tumour types.Conclusions:  In the majority of histologically suspected GISTs a combination of CD117 and DOG1 immunostaining is sufficient to confirm the histological diagnosis.
OsRAN2, essential for mitosis, enhances cold tolerance in rice by promoting export of intranuclear tubulin and maintaining cell division under cold stress
Authors: CHEN NA , XU YUNYUAN , WANG XIN , DU CHENG , DU JIZHOU , YUAN MING , XU ZHIHONG , CHONG KANG 
Published: September 2010

ABSTRACTWith global climate change, abnormally low temperatures have affected the world's rice production. Many genes have been shown to be essential for molecular improvement of rice cold‐tolerance traits. However, less is known about the molecular cellular mechanism of their response to cold stress. Here, we investigated OsRAN2 involved in regulation of cell division during cold stress in rice. Expression of OsRAN2 was increased under cold treatment, but not during salt and drought stress. The mean root mitotic index was closely related to the expression level of OsRAN2. Knockdown transgenic rice lines showed an aberrant organization of spindles during mitosis and stunted growth during development. Overexpression of OsRAN2 enhanced cold tolerance in rice. The transgenic rice overexpressing OsRAN2 showed maintained cell division, decreased proportion of cells with intranuclear tubulin and formation of a normal nuclear envelope under the cold condition. Our study suggests a mechanism for OsRAN2 in regulating cold resistance in rice by maintaining cell division through promoting the normal export of intranuclear tubulin at the end of mitosis. This insight could help improve the cold‐tolerance trait in rice.
Living donor liver transplantation using a graft from a donor with Dubin–Johnson syndrome
Authors: Liu Chinsu , Niu Dau‐Ming , Hsia Cheng‐Yuan , Loong Che‐Chuan , Lin Niang‐Cheng , Tsai Hsin‐Lin , Tsou Mei‐Yung , Chin Taiwai 
Published: September 2010

Liu C, Niu D‐M, Hsia C‐Y, Loong C‐C, Lin N‐C, Tsai H‐L, Tsou M‐Y, Chin T. Living donor liver transplantation using a graft from a donor with Dubin–Johnson syndrome.  Pediatr Transplantation 2012: 16: E25–E29. © 2010 John Wiley & Sons A/S.Abstract:  DJS is an autosomal recessive disorder that causes an increase in conjugated bilirubin without elevation of liver enzymes. Most patients are asymptomatic and have normal life spans, but to the best of our knowledge, their livers have never been reported to be grafts in liver transplantation. Herein, we report an infant patient with MMA that received a partial liver graft from his mother, who had DJS. A biliary anastomosis stricture was noted five months after transplantation and was successfully treated with radiological interventions. Otherwise, the patient’s liver functions were normal, and a liver biopsy showed a pathognomonic picture of DJS nine months after the transplantation. The patient was followed for one yr, and the results were satisfactory for an increase in oral intake and protein uptake, no recurrence of metabolic stroke and there was a gradual catch‐up with regard to physical development despite having a persistently abnormal profile of amino acid analysis. From the experience of our case, we suggest that a liver from a donor with DJS can be used as a graft for liver transplantation, although long‐term follow‐up is mandatory to examine the grafted liver under the use of immunosuppressive medications.
Derivation and Validation of a MEDLINE Search Strategy for Research Studies That Use Administrative Data
Authors: Van Walraven Carl , Bennett Carol , Forster Alan J. 
Published: September 2010

Objective. To derive and validate a search strategy that identifies administrative database research (ADR) in the MEDLINE database.Design. Analytical survey.Methods. We downloaded all articles published between January 1, 2008 and October 7, 2009 in 20 top journals in internal medicine, cardiovascular medicine, public health, and health services research. These were reviewed to determine whether they were ADR (in which the study cohort, exposure, or outcome was defined using electronic data created for or during the processing of patients through their health care). We used chi‐squared recursive partitioning to create a search strategy that maximized sensitivity based on publication type, MeSH headings, and text words.Main Outcome Measures. Sensitivity and positive predictive value of the search strategy for true ADR in three samples: derivation (n=5,513); internal validation (n=2,710); and external validation (n=1,500).Results. The prevalence of ADR in the derivation, internal validation, and external validation samples was 2.6, 2.9, and 2.2 percent, respectively. The sensitivity of our search strategy in these samples was 90.9 percent (95 percent confidence interval [CI] 85.0–95.1), 88.5 percent (79.2–94.6), and 100 percent (99.3–100), respectively. The positive predictive value in these samples was 10.7 percent (9.0–12.6), 11.5 percent (9.1–14.4), and 3.3 percent (2.3–4.6), respectively.Conclusion. We derived and validated a search strategy that is highly sensitive for ADR in MEDLINE.
Cancer-Associated Fibroblasts Enhance the Gland-Forming Capability of Prostate Cancer Stem Cells
Authors: Liao Chun-Peng , Adisetiyo Helty , Liang Mengmeng , Roy-Burman Pradip 
Published: September 2010

Abstract
Signals originating from cancer-associated fibroblasts (CAF) may positively regulate proliferation and tumorigenicity in prostate cancer. In this study, we investigated whether CAFs may regulate the biology of prostate cancer stem cells (CSC) by using a conditional Pten deletion mouse model of prostate adenocarcinoma to isolate both CAF cultures and CSC-enriched cell fractions from the tumors. CSCs that were isolated possessed self-renewal, spheroid-forming, and multipotential differentiation activities in tissue culture, segregating with a cell fraction exhibiting a signature expression phenotype, including SCA-1 (high), CD49f (high), CK5 (high), p63 (high), Survivin (high), RUNX2 (high), CD44 (low), CD133 (low), CK18 (low), and Androgen Receptor (low). CSC spheroid–forming efficiency was differentially influenced by the nature of fibroblasts in a coculture system: Compared with mouse urogenital sinus mesenchyme or normal prostate fibroblasts, CAFs enhanced spheroid formation, with the spheroids displaying generally larger sizes and more complex histology. Graft experiments showed that CSCs admixed with CAFs produced prostatic glandular structures with more numerous lesions, high proliferative index, and tumor-like histopathologies, compared with those formed in the presence of normal prostate fibroblasts. Together, our findings underscore a significant role of CAFs in CSC biology. Cancer Res; 70(18); 7294–303. ©2010 AACR.