bluuebunny/crossref_metadata_2025_split huggingface dataset:
High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle Virus
Authors: Lombardi Raffaele , Circelli Patrizia , Villani Maria Elena , Buriani Giampaolo , Nardi Luca , Coppola Valentina , Bianco Linda , Benvenuto Eugenio , Donini Marcello , Marusic Carla 
Published: November 2009

Abstract

Background
In recent years, different HIV antigens have been successfully expressed in plants by either stable transformation or transient expression systems. Among HIV proteins, Nef is considered a promising target for the formulation of a multi-component vaccine due to its implication in the first steps of viral infection. Attempts to express Nef as a single protein product (not fused to a stabilizing protein) in transgenic plants resulted in disappointingly low yields (about 0.5% of total soluble protein). In this work we describe a transient expression system based on co-agroinfiltration of plant virus gene silencing suppressor proteins in Nicotiana benthamiana, followed by a two-step affinity purification protocol of plant-derived Nef.

Results
The effect of three gene silencing viral suppressor proteins (P25 of Potato Virus X, P19 of either Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef transient expression yield was evaluated. The P19 protein of Artichoke Mottled Crinckle virus (AMCV-P19) gave the highest expression yield in vacuum co-agroinfiltration experiments reaching 1.3% of total soluble protein, a level almost three times higher than that previously reported in stable transgenic plants. The high yield observed in the co-agroinfiltrated plants was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms by AMCV-P19. Interestingly, we also showed that expression levels in top leaves of vacuum co-agroinfiltrated plants were noticeably reduced compared to bottom leaves. Moreover, purification of Nef from agroinfiltrated tissue was achieved by a two-step immobilized metal ion affinity chromatography protocol with yields of 250 ng/g of fresh tissue.

Conclusion
We demonstrated that expression level of HIV-1 Nef in plant can be improved using a transient expression system enhanced by the AMCV-P19 gene silencing suppressor protein. Moreover, plant-derived Nef was purified, with enhanced yield, exploiting a two-step purification protocol. These results represent a first step towards the development of a plant-derived HIV vaccine.
Cytotoxic and antibacterial activities of endophytic fungi isolated from plants at the National Park, Pahang, Malaysia
Authors: Hazalin Nurul AMN , Ramasamy Kalavathy , Lim Siong Siong Meng , Wahab Ibtisam Abdul , Cole Anthony LJ , Abdul Majeed Abu Bakar 
Published: November 2009

Abstract

Background
Endophytes, microorganisms which reside in plant tissues, have potential in producing novel metabolites for exploitation in medicine. Cytotoxic and antibacterial activities of a total of 300 endophytic fungi were investigated.

Methods
Endophytic fungi were isolated from various parts of 43 plants from the National Park Pahang, Malaysia. Extracts from solid state culture were tested for cytotoxicity against a number of cancer cell lines using the MTT assay. Antibacterial activity was determined using the disc diffusion method.

Results
A total of 300 endophytes were isolated from various parts of plants from the National Park, Pahang. 3.3% of extracts showed potent (IC50 < 0.01 μg/ml) cytotoxic activity against the murine leukemic P388 cell line and 1.7% against a human chronic myeloid leukemic cell line K562. Sporothrix sp. (KK29FL1) isolated from Costus speciosus showed strong cytotoxicity against colorectal carcinoma (HCT116) and human breast adenocarcinoma (MCF7) cell lines with IC50 values of 0.05 μg/ml and 0.02 μg/ml, respectively. Antibacterial activity was demonstrated for 8% of the extracts.

Conclusion
Results indicate the potential for production of bioactive agents from endophytes of the tropical rainforest flora.
Health care utilisation amongst Shenzhen migrant workers: does being insured make a difference?
Authors: Mou Jin , Cheng Jinquan , Zhang Dan , Jiang Hanping , Lin Liangqiang , Griffiths Sian M 
Published: November 2009

Abstract

Background
As one of the most populous metropolitan areas in the Pearl River Delta of South China, Shenzhen attracts millions of migrant workers annually. The objectives of this study were to compare health needs, self-reported health and healthcare utilisation of insured and uninsured migrant workers in Shenzhen, China, where a new health insurance scheme targeting at migrant workers was initiated.

Methods
A cross-sectional survey using multi-staged sampling was conducted to collect data from migrant factory workers. Statistical tests included logistic regression analysis were used.

Results
Among 4634 subjects (96.54%) who responded to the survey, 55.11% were uninsured. Disease patterns were similar irrespective of insurance status. The uninsured were more likely to be female, single, younger and less educated unskilled labourers with a lower monthly income compared with the insured. Out of 1136 who reported illness in the previous two weeks, 62.15% did not visit a doctor. Of the 296 who were referred for inpatient care, 48.65% did not attend because of inability to pay. Amongst those who reported sickness, 548 were insured and 588 were uninsured.
Those that were insured, and had easier access to care were more likely to make doctor visits than those who were uninsured.

Conclusion
Health care utilisation patterns differ between insured and uninsured workers and insurance status appears to be a significant factor. The health insurance system is inequitably distributed amongst migrant workers. Younger less educated women who are paid less are more likely to be uninsured and therefore to pay out of pocket for their care. For greater equity this group need to be included in the insurance schemes as they develop.
Oxaliplatin-Induced Loss of Phosphorylated Heavy Neurofilament Subunit Neuronal Immunoreactivity in Rat Drg Tissue
Authors: Jamieson Stephen MF , Subramaniam Joshuan , Liu Johnson J , Jong Nancy N , Ip Virginia , Connor Bronwen , McKeage Mark J 
Published: November 2009

Background Oxaliplatin and related chemotherapeutic drugs cause painful chronic peripheral neuropathies in cancer patients. We investigated changes in neuronal size profiles and neurofilament immunoreactivity in L5 dorsal root ganglion (DRG) tissue of adult female Wistar rats after multiple-dose treatment with oxaliplatin, cisplatin, carboplatin or paclitaxel. Results After treatment with oxaliplatin, phosphorylated neurofilament heavy subunit (pNF-H) immunoreactivity was reduced in neuronal cell bodies, but unchanged in nerve fibres, of the L5 DRG. Morphometric analysis confirmed significant changes in the number (-75%; P < 0.0002) and size (-45%; P < 0.0001) of pNF-H-immunoreactive neurons after oxaliplatin treatment. pNF-H-immunoreactive neurons had overlapping size profiles and co-localisation with neurons displaying cell body immunoreactivity for parvalbumin, non-phospho-specific neurofilament medium subunit (NF-M) and non-phospho-specific neurofilament heavy subunit (NF-H), in control DRG. However, there were no significant changes in the numbers of neurons with immunoreactivity for parvalbumin (4.6%, P = 0.82), NF-M (-1%, P = 0.96) or NF-H (0%; P = 0.93) after oxaliplatin treatment, although the sizes of parvalbumin (-29%, P = 0.047), NF-M (-11%, P = 0.038) and NF-H (-28%; P = 0.0033) immunoreactive neurons were reduced. In an independent comparison of different chemotherapeutic agents, the number of pNF-H-immunoreactive neurons was significantly altered by oxaliplatin (-77.2%; P < 0.0001) and cisplatin (-35.2%; P = 0.03) but not by carboplatin or paclitaxel, and their mean cell body area was significantly changed by oxaliplatin (-31.1%; P = 0.008) but not by cisplatin, carboplatin or paclitaxel. Conclusion This study has demonstrated a specific pattern of loss of pNF-H immunoreactivity in rat DRG tissue that corresponds with the relative neurotoxicity of oxaliplatin, cisplatin and carboplatin. Loss of pNF-H may be mechanistically linked to oxaliplatin-induced neuronal atrophy, and serves as a readily measureable endpoint of its neurotoxicity in the rat model.
[68Ga]-DOTATOC-PET/CT for meningioma IMRT treatment planning
Authors: Gehler Barbara , Paulsen Frank , Öksüz Mehmet Ö , Hauser Till-Karsten , Eschmann Susanne M , Bares Roland , Pfannenberg Christina , Bamberg Michael , Bartenstein Peter , Belka Claus , Ganswindt Ute 
Published: November 2009

Abstract

Purpose
The observation that human meningioma cells strongly express somatostatin receptor (SSTR 2) was the rationale to analyze retrospectively in how far DOTATOC PET/CT is helpful to improve target volume delineation for intensity modulated radiotherapy (IMRT).

Patients and Methods
In 26 consecutive patients with preferentially skull base meningioma, diagnostic magnetic resonance imaging (MRI) and planning-computed tomography (CT) was complemented with data from [68Ga]-DOTA-D Phe1-Tyr3-Octreotide (DOTATOC)-PET/CT. Image fusion of PET/CT, diagnostic computed tomography, MRI and radiotherapy planning CT as well as target volume delineation was performed with OTP-Masterplan®. Initial gross tumor volume (GTV) definition was based on MRI data only and was secondarily complemented with DOTATOC-PET information. Irradiation was performed as EUD based IMRT, using the Hyperion Software package.

Results
The integration of the DOTATOC data led to additional information concerning tumor extension in 17 of 26 patients (65%). There were major changes of the clinical target volume (CTV) which modify the PTV in 14 patients, minor changes were realized in 3 patients. Overall the GTV-MRI/CT was larger than the GTV-PET in 10 patients (38%), smaller in 13 patients (50%) and almost the same in 3 patients (12%). Most of the adaptations were performed in close vicinity to bony skull base structures or after complex surgery. Median GTV based on MRI was 18.1 cc, based on PET 25.3 cc and subsequently the CTV was 37.4 cc. Radiation planning and treatment of the DOTATOC-adapted volumes was feasible.

Conclusion
DOTATOC-PET/CT information may strongly complement patho-anatomical data from MRI and CT in cases with complex meningioma and is thus helpful for improved target volume delineation especially for skull base manifestations and recurrent disease after surgery.
Cecum perforation due to tuberculosis in a renal transplant recipient: a case report
Authors: Carkman Sinan , Ozben Volkan , Aytac Erman 
Published: November 2009

Abstract

Introduction
Tuberculosis can present in many varied clinical situations in immunosuppressed patients. It has been reported that the sigmoid colon is the most common site for colonic perforation in renal transplant recipients and diverticulitis is its most common cause. Cecal perforation because of tuberculosis is extremely rare in a renal transplant recipient. We present the case of a renal transplant patient with cecal perforation due to tuberculosis, 10 years after renal transplantation.

Case presentation
A 39-year-old Caucasian man, who was a renal transplant recipient, was admitted to our emergency surgery unit with an acute abdomen. A cecal perforation was found at exploratory laparotomy, and a right hemicolectomy with an end ileostomy and transverse colonic mucous fistula were performed. Necrotizing granulomatous colitis due to tuberculosis was reported in the histopathologic examination.

Conclusion
Colonic perforations in immunosuppressed patients may have unusual presentations and unusual causes. Tuberculosis infection should be considered in the differential diagnosis during the histopathologic evaluation in immunocompromised patients such as renal transplant recipients.
CTCF binding site classes exhibit distinct evolutionary, genomic, epigenomic and transcriptomic features
Authors: Essien Kobby , Vigneau Sebastien , Apreleva Sofia , Singh Larry N , Bartolomei Marisa S , Hannenhalli Sridhar 
Published: November 2009

Abstract
Background
CTCF (CCCTC-binding factor) is an evolutionarily conserved zinc finger protein involved in diverse functions ranging from negative regulation of MYC, to chromatin insulation of the beta-globin gene cluster, to imprinting of the Igf2 locus. The 11 zinc fingers of CTCF are known to differentially contribute to the CTCF-DNA interaction at different binding sites. It is possible that the differences in CTCF-DNA conformation at different binding sites underlie CTCF's functional diversity. If so, the CTCF binding sites may belong to distinct classes, each compatible with a specific functional role.

Results
We have classified approximately 26,000 CTCF binding sites in CD4+ T cells into three classes based on their similarity to the well-characterized CTCF DNA-binding motif. We have comprehensively characterized these three classes of CTCF sites with respect to several evolutionary, genomic, epigenomic, transcriptomic and functional features. We find that the low-occupancy sites tend to be cell type specific. Furthermore, while the high-occupancy sites associate with repressive histone marks and greater gene co-expression within a CTCF-flanked block, the low-occupancy sites associate with active histone marks and higher gene expression. We found that the low-occupancy sites have greater conservation in their flanking regions compared to high-occupancy sites. Interestingly, based on a novel class-conservation metric, we observed that human low-occupancy sites tend to be conserved as low-occupancy sites in mouse (and vice versa) more frequently than expected.

Conclusions
Our work reveals several key differences among CTCF occupancy-based classes and suggests a critical, yet distinct functional role played by low-occupancy sites.
Investigation of force-controlled friction stir welding for manufacturing and automation
Authors: Longhurst W R , Strauss A M , Cook G E , Cox C D , Hendricks C E , Gibson B T , Dawant Y S 
Published: November 2009

Friction stir welding (FSW) is a solid-state joining process for materials with low melting points. The process uses a rotating tool that consists of a shoulder and a pin. The tool plastically deforms the material with its pin and then forges together the parent materials underneath the shoulder. Past research has established that the axial force on the tool that creates the forging pressure is a function of plunge depth, traverse speed, and rotation speed. Historically, force control of FSW has been accomplished by varying the plunge depth of the tool. The research presented in this paper examines the force control of FSW by varying each of the process parameters separately. A force controller was implemented on a retrofitted milling machine. The closed-loop proportional—integral—derivative (PID) control architecture was tuned using the Ziegler—Nichols method. Welding experiments were conducted by butt welding ¼ inch (6.35 mm) × 1½ inch (38.1 mm) × 8 inch (203.2 mm) samples of aluminium 6061-T6511 with a ¼ inch (6.35 mm) FSW tool. The results indicate that force control via traverse speed is the most accurate and, as a by-product, heat distribution control along the weld seam occurs. Force control via plunge depth is the least accurate but it compensates for machine and robot deflection. Tensile test data show that greater strength can be obtained through force control via rotation speed. It is concluded that force is maintained by keeping the amount of tool surface area in contract with the workpiece constant throughout the welding process when plunge depth is used as the controlling variable. Force is maintained by varying the rate of heat generation when rotation speed is used as the controlling variable. Lastly, force is maintained by changing the amount of heat deposited per unit length along the weld seam when traverse speed is used as the controlling variable. Successful robotic FSW requires to be selected the appropriate controlling variable and the sensitivity of the interaction between the tool and the workpiece to be reduced.
Multigenic combination of estrogen-related genes is associated with age at natural menopause in a Spanish population
Authors: Mendoza Nicolás , Sánchez-Borrego Rafael , Galiano Daniela , Salamanca Alberto , Mozas Juan , Quereda Francisco , Vázquez Francisco , Martínez-Astorquiza Txantón , Moron Francisco 
Published: November 2009

Objective Age at natural menopause (ANM) can be considered a complex parameter that depends on the interaction of multiple factors. In the present study, the role of interaction between genetic variants within estrogen synthesis and signalling pathways in the ANM in Spanish women is studied. Material and methods Nine single nucleotide polymorphisms (SNPs) located at different candidate genes related to the estrogen signalling pathway were analysed in 1980 Spanish postmenopausal women. Results Independently, none of the nine markers were significantly associated with early ANM. Only heterozygosis at the NRIP rs2229741 locus could be associated with early menopause; however, this marker does not maintain statistical significance. In contrast, linear regression analysis suggests several epistatic interactions including these markers in relation to ANM, especially between ESR2, NRIP1 and BMP15. The genetic variant that appears most in these interactions is that of the BMP15 rs3897937. It was observed that AA-TC combined genotype for NRIP-BMP15 (rs3897937), respectively, appears to be associated with a lower ANM than other possible combinations of these SNP (46.1±5.9 versus 50.4±3.3; P = 0.002). In the multilocus analysis, the multigenic interaction formed by ESR2 (AA), BMP15 rs3897937 (TC) and NRIP1 (AA) has the lower ANM (45.37±6.8 versus 48.69±5; P = 0.038). Conclusions The results suggest that epistatic interactions of estrogen-related alleles may contribute to variance in ANM in Spanish women. Moreover, BMP15 and NRIP1 also appear as attractive candidate genes for premature menopause but require further investigation to confirm them.
Pilot Study of Linking Web-Based Supplemental Interpretive Information to Laboratory Test Reports
Authors: Shirts Brian H. , Gundlapalli Adi V. , Jackson Brian 
Published: November 2009

Abstract
Electronic medical records have the ability to link to reference material, providing clinicians with immediate access to information relevant to patient care. Adding relevant links to laboratory test results could add value while minimizing the volume of ancillary text presented.
We provided Web-based universal resource locator (URL) links with all results of 7 laboratory tests ordered at ARUP Laboratories (Salt Lake City, UT). URL links provided were modified 7 months later, and use between initial and subsequent URLs was tracked to establish frequency and duration of access to supplemental Web information.
Monthly Web-site hit rates for individual tests varied from 0.00% to 3.00% (median, 0.12%). Rare and specialty tests averaged higher hit rates. There was no decay in hit rate 9 months after URLs were removed from test reports.
We conclude that links to reference material are accessed by clinicians. The use of Web links months after links were no longer published raises an important issue of long-term maintenance and the resources required to support these features.
Progress
Authors: Walshe John 
Published: November 2009

On returning to University College Hospital in London I needed to find patients with this rare disease and supplies of penicillamine to assess its ability to mobilise copper from their abnormal body stores. Three patients were found by my father, Sir Francis Walshe, a distinguished neurologist, and with the small amount of penicillamine available to me I was able to show that in each case penicillamine promoted the excretion of more copper than did BAL. Two patients were returned to their referring physicians but the third stayed with me for over 50 years and made an excellent recovery eventually having three children of her own.
Supplies of penicillamine were secured when I was able to convince the medical director of the Distillers Company, the principal makers of penicillin at the time, to make penicillamine for me.

By the early 1960s reports from my own work and from other centres appeared showing that patients treated with penicillin showed a remarkable improvement in all symptoms.
However Propfessor Denny Brown and Dr Uzman in Boston still believed that Wilson's disease was not due to copper deposition but due to abnormal protein metabolism with abnormal peptides being excreted in the urine. Working with Professor Milne and Dr Asatoor at the Hammersmith Hospital, it was possible to disprove this theory using newer techniques to analyse urine.
In 1957, after failing to get appointed to two different appointments in Oxford I was accepted to a post in the Department of Experimental Medicine in Cambridge.
A New Beginning
Authors: Walshe John 
Published: November 2009

Having been appointed Assistant Director of Research to Professor McCance in Cambridge I needed to find somewhere to live and to decide upon a line of research in keeping with the facilities available.. Shortly after taking up this new post I was invited, by the British Council, to undertake a lecture tour in South America. On returning from this I succeeded in establishing the methods I needed to estimate abnormalities of copper metabolism and to recruit patients for my studies. I also continued my work with Sydney Osborn, a medical physicist, and we improved our methods of determining the movement of copper in the body using a short half life radioactive isotope of copper.
Whilst the results of treatment with penicillamine were very encouraging I encountered the problem of a patient developing a severe toxic reaction to the drug. A University biochemist, Dr Hal Dixon, suggested I use triethylenetetramine, a known copper binding compound and this proved safe and effective. For several years we produced this in my laboratory to treat a small number of penicillamine intolerant patients. We also found that an impurity in this compound could induced a disastrous fall in blood pressure. During the course of these studies we moved premises twice, first temporary war time huts in Canhams yard and then the old Low Temperature Research building in the Downing Street site. During this time I also had trouble with the Ministry of Health over the question of prescription charges for my patients.
The Department of Experimental Medicine
Authors: Walshe John 
Published: November 2009

The story resumes with yet another move, the third in all, to new laboratories in the University wing at the New Addenbrooke's Hospital. This had the advantage of being near my ward, the service departments, biochemistry, haematology, radiology and audiovisual aids. It had the disadvantage of removing me from the basic science departments of the University with whom I had had such successful cooperation. In addition my new quarters were much smaller than before and there was no convenient place to see patients. The major problem was the roof. Every time it rained heavily the roof leaked and I had to take unusual action to get this remedied.
However the main problem was the realisation that the impurity in our trientine preparation was not only potentially toxic but, perhaps actually so. This lead to a search for and, eventually thanks again to Hal Dixon, the finding of a solution to the problem. Other activities were a search for a common ancestor to a cluster of Wilson patients centred around the Wash; the search through church records was fascinating but, unfortunately, non productive; the cluster must have been coincidental There was also some work for an ice cream firm who allowed a batch of iced lollies with excess copper onto the market. Finally there was continued work on basic brain chemistry with Sir Rudolph Peters in the University Department of Biochemistry in which we showed how copper might damage the transport of ions through the neuronal membrane........