bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Interlaminar Epidural Steroid Injection in Acute and Subacute Pain Due to Lumbar Disk Herniation: A Randomized Comparison of 2 Different Protocols
Authors: Gelalis I.D. , Arnaoutoglou E. , Pakos E.E. , Politis A.N. , Rapti M. , Xenakis T.A. , Papadopoulos G.
Published: January 2010

In order to assess the efficacy of epidural steroid injections (ESI) in acute and subacute pain due to lumbar spine disk herniation, we conducted a randomized trial, comparing 2 different protocols. Fourty patients with radicular pain due to L4-L5 and L5-S1 disc herniation were assigned to receive either 3 consecutive ESI every 24 hours through a spinal catheter (group A) or 3 consecutive ESI every 10 days with an epidural needle (group B). All patients had improved Oswestry Disabilty Index (ODI) and the Visual Analog Scale (VAS) for pain scores at 1 month of follow-up compared to baseline, while no significant differences were observed between the 2 groups. The scores for group B were statistically significant lower at 2 months of follow-up compared to those of group A. The improvement in the scores of group B was continuous since the mean scores at 2 months of follow up were lower compared to the respective scores at 1 month. Protocol B (3 consecutive ESI every 10 days) was found more effective in the treatment of subacute pain compared to Protocol A (3 consecutive ESI every 24 hours) with statistically significant differences in the ODI and VAS scores at 2 months of follow-up.
Sciatic Neuroma Presenting Forty Years After Above-Knee Amputation
Authors: Kitcat M. , Hunter J.E. , Malata C.M.
Published: January 2010

We report a case of a sciatic neuroma presenting forty years after above knee amputation. Patients developing neuroma following a limb amputation can present with stump pain which is commonly resistant to medical intervention. The length of interval from the initial injury to presentation is widely variable. Diagnosis relies on clinical suspicion and accurate assessment, radiological imaging and, if indicated, surgical exploration. MRI provides a better soft tissue definition than CT and is more accurate in identifying small lesions than ultrasound. The aim of treatment for symptomatic neuroma is pain relief and improvement of function. This is often achieved by surgical excision.
Short and Long-Term Incidence of Tuberculosis and CD4-Cell Count Dynamic on HAART in Senegal
Authors: François Etard Jean- , Assane Diouf , Pierre De Beaudrap , Akoi Koivugui , Fatou Ngom-Guèye Ndèye , Ibrahima Ndiaye , René Ecochard , Salif Sow Papa , Eric Delaporte
Published: January 2010

Objectives: Estimate tuberculosis (TB) incidence among patients receiving HAART. Compare the dynamic of the CD4-cell count and viral load before notification of the TB with the dynamic among patients remaining free of TB.

Design: Prospective cohort with ascertainment of TB cases from medical records.

Methods: The first 404 adults HIV-1 infected patients enrolled in the Senegalese antiretroviral drug access initiative were eligible. CD4-cell and viral load were assessed at baseline and every 6 months. Patients receiving an antituberculosis treatment at HAART initiation were excluded from analysis. Any TB case notified after the first month of HAART was considered as an incident case. Follow-up was censored at death or at the last visit before March 31, 2008. CD4-cell trajectories until TB notification were compared to non-TB developers within two distinct periods: from HAART initiation to 24 months and after.

Results: Over 404 eligible patients, 352 were included in this analysis. Median follow-up reached 73 months and 1821 person-years were accrued. Half of the 42 incident cases were notified before month 19 of HAART yielding to an overall incident rate of 2.3/100 PY [1.7-3.1]. Annual incidence decreased with duration of HAART (trend in incidence: RR=0.26, p<10-4). During the first period, CD4-cell count dynamic of most TB patients was identical to the dynamic among patients remaining free of TB. Most cases of the second period occurred in a context of an immunological failure.

Conclusions: This study provides an estimate of TB incidence among patients on HAART in Senegal and supports two underlying mechanisms.
Pathology and Molecular Biology of Renal Neoplasms: Recent Advances and Impacts on Pathological Classification
Authors: Nagashima Yoji , Kobayashi Naohito , Kagawa Eriko , Aoki Ichiro , Inayama Yoshiaki , Yao Masahiro
Published: January 2010

Although the incidence of renal neoplasms is not particularly high, their clinical presentation is
often interesting. They tend to arise in association with familial neoplastic syndromes, i.e. renal cell
carcinoma with von Hippel-Lindau disease. The cloning of the genes responsible for these hereditary
pathological conditions revealed their involvement in sporadic renal cancers. Based on their molecular
biological features, the classification of renal neoplasms was reassessed by the World Health Organization in
2004. In this chapter we discuss the relationships between the pathology and molecular biology of each
histological type of renal neoplasm and the underlying molecular biological abnormalities.
Chemotherapy in Malignant Gliomas
Authors: Nakamura Mitsutoshi , Shimada Keiji , Nakase Hiroyuki , Konishi Noboru
Published: January 2010

The current practice of relying solely on microscopic examinations for histological classification of gliomas and, consequentially, determination of optimal treatment, appears to be insufficient. As a result of the increasing use of molecular markers in tumor classification, there is an emerging emphasis in the genetic profiles of distinct subtypes of glioma. Glioblastomas classified as WHO grade IV are the most malignant astrocytic tumors and may develop either as de novo primary glioblastomas or as secondary glioblastomas arising from lower-grade astrocytomas. Both primary and secondary glioblastomas show similar histological features, but these subtypes constitute molecularly distinct entities evolving through different genetic pathways and likely to differ in prognosis and in response to therapy. The standard of care involves postoperative radiotherapy with concomitant and sequential administration of temozolomide (TMZ); however, the methylation status of O6-methylguanine-methyltransferase (O6-MGMT) appears to relate to TMZ resistance. Oligodendroglioma (WHO grade II) and anaplastic oligodendroglioma (WHO grade III) are particular glioma subtypes, derived from oligodendrocytes, that show remarkable response to a specific chemotherapy regimen of procarbazine + CCNU + vincristine (PCV), making their correct diagnosis important. Without the availability of typical morphological features, however, histological differentiation of oligodendrogliomas from astrocytoma is highly subjective. Inclusion of such molecular markers – for example, loss of heterozygosity (LOH) on chromosomes 1p and 19q, which is correlated with sensitivity to PCV chemotherapy and with increased survival in patients specifically with oligodendroglial tumors - will help to tailor treatment to the individual patient. This article reviews biological and molecular approaches to glioma classification that have the potential to increase the efficacy of treatments for these tumors.
Thyroid Cancer: Gene Mutation and Morphology of Follicular Cell Carcinomas
Authors: Nakamura Misa , Li Yaqiong , Liu Zhiyan , Kakudo Kennichi
Published: January 2010

Thyroid carcinoma is the most prevalent type of endocrine cancer and its incidence has
reportedly increased significantly over the last few decades in many areas of the world. The traditional
classification of thyroid carcinoma divides it into four common types, including papillary, follicular,
medullary and anaplastic carcinomas. In recent years, much progress has been made regarding the
molecular characterization of these carcinomas, clarifying the genetic alterations underlying the histotype
diversity of these types of cancer. The most common mutations in thyroid cancer are point mutations of
the BRAF, RAS, p53 and β-catenin genes, and RET/PTC and PAX8-PPARγ rearrangements. Many of
these mutations, particularly those leading to activation of the MAPK pathway, are being actively
explored as therapeutic targets for thyroid cancer. In this review we have summarized these specific
genetic changes in the process of development and dedifferentiation in thyroid cancer derived from
thyroid follicular cells.
Consistency of Diagnostic Thresholds in DSM-V
Authors: Andrews Gavin , Sunderland Matthew , Kemp Alice
Published: January 2010

Objective: DSM-IV diagnostic criteria define thresholds on a continuum of symptoms above which the diagnosis is said to be established. Data from the 1997 Australian Survey of Mental Health and Wellbeing were used for six internalizing disorders, and the levels of distress and disability associated with each diagnosis were investigated.

Method: Mean distress (measured by the K-10) and disability (measured by the SF12-MCS) scores were identified for people in the Survey who reported no physical or mental disorders. The distribution of distress and disability showed by people who met criteria for major depressive disorder, dysthymia, generalized anxiety disorder, social phobia, post-traumatic stress disorder and obsessive–compulsive disorder was plotted against the mean for well people, expecting that ≥90% of people with these mental disorders would score as more distressed or disabled than this mean.

Results: More than 90% of people with dysthymia, major depressive disorder, generalized anxiety disorder or with post-traumatic stress disorder scored as more distressed or disabled than the mean for well people. A majority were severely distressed or disabled (>2SD above the mean). This remained the case when the clinical significance criteria were removed. In social phobia and in obsessive–compulsive disorders between 9% and 26% scored below the means for well people, that is, as neither distressed nor disabled, a figure that rose to 16–40% when the clinical significance criteria were removed. In neither case did a majority of cases score in the severe range.

Conclusions: The diagnostic thresholds for social phobia and for obsessive–compulsive disorder are less stringent than that for the other disorders and require revision in DSM-V.
Sex Differences in Neuropsychological Functioning Among Schizophrenia Patients
Authors: Bozikas Vasilis P. , Kosmidis Mary H. , Peltekis Apostolos , Giannakou Maria , Nimatoudis Ioannis , Karavatos Athanasios , Fokas Kostas , Garyfallos George
Published: January 2010

Objectives: Evidence from the literature addressing sex differences in cognition in schizophrenia remains equivocal, with some researchers suggesting that male schizophrenia patients are more impaired than female subjects, while others report no significant sex differences in cognitive functioning. The aim of the present study was to investigate whether the differential pattern of cognitive performance observed in healthy men and women is preserved in male and female schizophrenia patients.

Method: Ninety-six schizophrenia patients (56 men) were compared with 62 age- and gender-ratio matched healthy controls (31 men), on a battery of neuropsychological tests that assessed basic cognitive abilities: attention, working memory, abstraction, inhibition, fluency, verbal learning and memory, visual memory, visuospatial skills, and psychomotor speed.

Results: As a group, schizophrenia patients were significantly impaired in each of the cognitive domains assessed, with the exception of psychomotor speed. The effect of sex was significant for verbal learning and memory, wherein women outperformed men. No significant group × sex interactions were found in any cognitive domains, indicating that the female advantage typically observed in verbal learning and memory remained the same in the schizophrenia patients.

Conclusion: The degree of cognitive impairment is the same for male and female schizophrenia patients. Those sex differences found among the patients were typical of the healthy population as well. Therefore, differential decrements in basic cognitive domains do not appear to account for the favourable course of schizophrenia in women relative to men.
Metadata for Name Disambiguation and Collocation
Authors: Beall Jeffrey
Published: January 2010

Searching names of persons, families, and organizations is often difficult in online databases because different persons or organizations frequently share the same name and because a single person’s or organization’s name may appear in different forms in various online documents. Databases and search engines can use metadata as a tool to solve the problem of name ambiguity and name variation in online databases. This article describes the challenges names pose in information retrieval and some emerging name metadata databases that can help ameliorate the problems. Effective name disambiguation and collocation increase search precision and recall and can improve assessment of scholarly work.
Detection of Multidecadal Changes in UVB and Total Ozone Concentrations over the Continental US with NASA TOMS Data and USDA Ground-Based Measurements
Authors: Gao Zhiqiang , Gao Wei , Chang Ni-Bin
Published: January 2010

Thinning of the atmospheric ozone layer leads to elevated levels of Ultraviolet-B (UVB) at the Earth's surface, resulting in an increase of health risks to living organisms due to DNA damage. This paper examines the multidecadal changes of total column ozone from 1979 to 2005 with the aid of ground-based UVB stations using the ultraviolet multifilter rotating shadow-band radiometer (UV-MFRSR). For the purpose of demonstration, four USDA ground stations, WA01, CO01, MD01, and AZ01, were selected for detailed comparisons against the satellite data. The major finding of this study is that over the course of the time series, on a monthly scale, the UV index (UVI) has increased at the four selected USDA stations while total ozone has decreased in the continental USA over the past three decades and spatial distributions of UVI and total ozone have shown substantial variations from coastal zones to the Midwest Regions of the USA, yet the tendency toward recovery of ozone layer in the continental USA cannot be fully confirmed. This leads to a conclusion that the UVI changes might have been influenced by other factors in addition to the total ozone in the atmospheric environment across at least 76% of the continental USA.
Binary Fingerprints at Fluctuation-Enhanced Sensing
Authors: Chang Hung-Chih , Kish Laszlo  B. , King Maria  D. , Kwan Chiman
Published: January 2010

We have developed a simple way to generate binary patterns based on spectral slopes in different frequency ranges at fluctuation-enhanced sensing. Such patterns can be considered as binary "fingerprints" of odors. The method has experimentally been demonstrated with a commercial semiconducting metal oxide (Taguchi) sensor exposed to bacterial odors (Escherichia coli and Anthrax-surrogate Bacillus subtilis) and processing their stochastic signals. With a single Taguchi sensor, the situations of empty chamber, tryptic soy agar (TSA) medium, or TSA with bacteria could be distinguished with 100% reproducibility. The bacterium numbers were in the range of 2.5 × 104-106. To illustrate the relevance for ultra-low power consumption, we show that this new type of signal processing and pattern recognition task can be implemented by a simple analog circuitry and a few logic gates with total power consumption in the microWatts range.
On Prerequisites for the Application of Sustainable Development Indicators in Urban Water Management
Authors: Palme Ulrika
Published: January 2010

Semi-structured interviews with 47 key actors were conducted in Swedish water utilities on why Sustainable Development Indicators (SDIs) are or are not used. Important influencing aspects identified included organizational inertia, social capital, the national water sector and authorities. Divergent views of SD and indicators appear to hinder SDI initiatives. Possible explanations are that: (a) not all actors look at decision-making as the kind of rational process the focus on indicators implies, and (b), Swedish urban water systems are widely regarded as sustainable. The water sector itself and regulation are identified as the strongest potential drivers for increased use of SDIs.
The False Cocoon of Hyposoter parorgyiae (Vier.) (Hymenoptera: Ichneumonidae)
Authors: Finlayson Thelma
Published: January 2010

Hyposoter parorgyiae (Vier.) (Hymenoptera: Ichneumonidae), a parasite that emerges from the mature larva of Parorgyia plagiata (Wlk.) (Lepidoptera: Liparidae), was found to have a false cocoon. The twelve specimens examined were collected in Ontario and Quebec, and are in the Canadian National Collection, Ottawa, Ontario.
Les Formicides (Hyménoptères) de Provancher
Authors: Francoeur A. , Béique R.
Published: January 2010

Toutes les espèces de Fourmis que Provancher a décrites sont tombées en synonymie. Cependant l'examen des types démontre que trois espèces s’avéraient nouvelles à l'époque. L’une d’entre elles fut désignée par un homonyme primaire. Nous proposons pour les deux autres une synonymie et une combinaison nouvelles.
High Temperature Mortality of the Sugar-pine Cone Beetle, Conophthorus lambertianae Hopkins (Coleoptera: Scolytidae)
Authors: Bedard William D.
Published: January 2010

Conophthorus lambertianae broods within cones in sunny places on the ground can be killed by high temperatures. This conclusion is supported by experimental evidence from four sources: (1) mortality of broods in cones placed in the sun; (2) mortality of cone beetles exposed to high temperatures in the laboratory; (3) measurements of internal cone temperatures occurring in nature; and (4) consideration of some factors that influence internal cone temperatures.
Development, Survival and Reproduction of Camnula pellucida (Scudder) (Orthoptera: Acrididae) in Relation to Climatic Conditions
Authors: Pickford R.
Published: January 2010

Field-rearing studies of Camnula pellucida (Scudder) conducted at Saskatoon from 1960 to 1962 indicated that marked differences often occurred in rate of development, survival and fecundity because of differing weather patterns. Grasshoppers usually developed more rapidly as they hatched later in the season when temperatures were rising. In addition, striking differences occurred in growth rates of grasshoppers hatched on similar dates in different years due to sharply contrasting climatic conditions. Although survival of nymphs was generally high, mortality was consistently greatest during early nymphal life. Precipitation or excessively hot weather during this period increased mortality considerably, yet did not affect the older nymphs. Survival of adults was also generally high whether they fledged early or late in the season. They were able to endure very high temperatures during mid-summer or below freezing temperatures during late autumn. The three-year average reproductive capacity of this grasshopper was approximately eight egg-pods or 176 eggs per female. The rate of oviposition varied with temperature, reaching a maximum per female of approximately one pod every four days during mid-summer when temperatures were high and declining thereafter as temperatures dropped. The average egg-pod contained 22.13 eggs which showed a tendency to increase as adults fledged later in the season.

It is suggested that climatic conditions frequently play a decisive role in the short-term modification of grasshopper infestations as well as in the long-term cyclic rise and fall in populations; consistently favourable or unfavourable conditions throughout the grasshopper cycle could result in an explosive outbreak or in almost complete elimination of any threat to the crop.
Ground Beetles (Coleoptera: Carabidae) in Relation to Agricultural Crops
Authors: Rivard I.
Published: January 2010

A three-year study on the seasonal occurrence of ground beetles was conducted in five fields with different crop rotations. More than 10,000 individuals representing 102 species were captured in pitfall traps. The relatively high total number of species collected the first year probably resulted from the particular location of the traps along field margins and close to woodlands. The variation observed between the areas studied was not necessarily related to the kind of crop. A progressive augmentation in beetle activity and population appeared to coincide with an increase in the humidity of the habitats and with the cultures in the following order: pasture grass, forage, cultivated, and cereal crops. This indicates that the nature of the crop influences beetle numbers, and from this it may he inferred that it also influences the extent to which the beetles exert a control influence on pest species. Of the eight species present every year and found in practically every field, Pterostichus melanarius Ill., Harpalus pennsylvanicus DeG., and H. compar Lec. were the most numerous.
Some Effects of Female Parasite Size on Reproduction ofNasonia vitripennis(Walk.) (Hymenoptera: Pteromalidae)
Authors: Wylie H. G.
Published: January 2010

Small females of the parasiteNasonia vitripennis(Walk.) parasitized fewer hosts (housefly,Musca domesticaL.) than larger ones during their lifetime because they had both a shorter life span and a lower rate of parasitization. The latter resulted primarily from a lower egg maturation rate. As females reared from superparasitized hosts are on the average smaller than those reared from non-superparasitized ones, these effects of parasite size reduce the rate of increase ofN.vitripennisat high parasite:host ratios and result in greater stability in the interaction between the two species.
Bumblebees From Northern Ellesmere Island, With Observations on Usurpation by Megabombus hyperboreus (Schönh.) (Hymenoptera: Apidae)
Authors: Milliron H. E. , Oliver D. R.
Published: January 2010

Two species of Apidae occur on Ellesmere Island: Megabombus (Megabombus) polaris (Curtis) and M. (M.) hyperboreus (Schönherr). Some observations on their activity are presented. Three nests built on the surface of the ground are described. The colonies were of small size. Nest No. 1 contained only polaris; Nests Nos. 2 and 3 contained polaris workers and hyperboreus sexuals, as adults or brood. Usurpation is suggested to explain the workerless condition of hyperboreus in Nests Nos. 2 and 3.
Sharing space: can ethnoprimatology contribute to the survival of nonhuman primates in human‐dominated globalized landscapes?
Authors: Lee P.C.
Published: January 2010

The emerging discipline of ethnoprimatology has at its core the construct that humans and nonhuman primates share a planet, an evolutionary history and a “primate perspective” on the world; more simply stated ethnoprimatolgy suggests that humans have perspectives on nonhuman primates which can contribute positively to the primates' enduring survival in our increasingly human‐dominated landscapes. Here, I explore whether humans can or do contribute positively to the conservation of nonhuman primates, or whether humanity's impact on, as well as our perceptions of, primates are generally negative. I examine primate–human interactions at the intersection of agriculture with natural habitats as exemplified in several long‐term studies, and explore the conservation consequences of these interactions. These interactions are then placed into an ecological‐economic perspective assessing the prospects for the survival of primates in a context where humans share their subsistence space and resources with primates. Am. J. Primatol. 72:925–931, 2010. © 2010 Wiley‐Liss, Inc.
Dissolved oxygen and pH profile evolution after cryovial thaw and repeated cell passaging in a T‐75 flask
Authors: Vallejos Jose R. , Brorson Kurt A. , Moreira Antonio R. , Rao Govind
Published: January 2010

Routine cell culture is done in small‐scale disposable vessels (typically 0.1–100 mL volumes) in academia and industry. Despite their wide use in bioprocess development (i.e., process optimization and process validation), miniature process scouting devices (PSDs) are considered “black boxes” because they are generally not equipped with sensors. In this study, we show that on‐line monitoring of dissolved oxygen (DO) and pH in a T‐75 flask‐based PSD can be achieved during cell passaging and that this information can be linked to different cellular metabolic states. In this case, on‐line monitoring of DO and pH show three distinctive metabolic regions in passages 1–18, 19–28, 29–54 and in particular, the shift in the pH curve, the specific oxygen uptake rate (qO2), and the lactate production rate to the oxygen consumption rate yield (YLac/ox) confirm the existence of these distinctive metabolic regions. These findings are particularly useful because they show that sensor equipped PSDs can help to monitor cell culture behavior after thaw, in pre‐ and seed culture prior to scale‐up and in development/optimization studies. Such routine monitoring will help to develop more consistent cell culture techniques. Biotechnol. Bioeng. 2010;105: 1040–1047. © 2009 Wiley Periodicals, Inc.
Influence of the hydrostatic pressure and pH on the asymmetric 2‐hydroxyketone formation catalyzed by Pseudomonas putida benzoylformate decarboxylase and variants thereof
Authors: Berheide Marco , Peper Stephanie , Kara Selin , Long Wei Sing , Schenkel Steffen , Pohl Martina , Niemeyer Bernd , Liese Andreas
Published: January 2010

Benzoylformate decarboxylase (BFD) from Pseudomonas putida is a thiamine diphosphate‐dependent (ThDP) enzyme that catalyzes the asymmetric CC bond formation to (S)‐2‐hydroxypropiophenone [(S)‐HPP] starting from benzaldehyde and acetaldehyde. The enantioselectivity of BFD was shown to be a function of temperature and substrate concentration. It can additionally be changed by site‐directed mutagenesis on hot spot positions in the active site. In this article, we present the effect of hydrostatic pressure up to 250 MPa on the enantioselectivity for the recombinant wtBFD as well as for the variants BFD F464I, BFD A460I, and BFD A460I‐F464I. A general tendency toward lower amounts of (S)‐HPP could be observed at increasing pressures. For two of these variants an increase in pressure even caused an inversion in the enantioselectivity and thus increasing enantiomeric excesses, respectively. A pressure‐induced increase in enantioselectivity could therefore be observed for the first time in biocatalysis to the best of our knowledge. Furthermore, the pH is shown to be a parameter that also significantly influences the enantioselectivity of the reaction mentioned above. Biotechnol. Bioeng. 2010; 106: 18–26. © 2009 Wiley Periodicals, Inc.
Disulfide trapping of protein complexes on the yeast surface
Authors: Lim Kok Hong , Madabhushi Sri Ranganayaki , Mann Jasdeep , Neelamegham Sriram , Park Sheldon
Published: January 2010

Protein complexes are common in nature and play important roles in biology, but studying the quaternary structure formation in vitro is challenging since it involves lengthy and expensive biochemical steps. There are frequent technical difficulties as well with the sensitivity and resolution of the assays. In this regard, a technique that can analyze protein–protein interactions in high throughput would be a useful experimental tool. Here, we introduce a combination of yeast display and disulfide trapping that we refer to as stabilization of transient and unstable complexes by engineered disulfide (STUCKED) that can be used to detect the formation of a broad spectrum of protein complexes on the yeast surface using fluorescence labeling. The technique uses an engineered intersubunit disulfide to covalently crosslink the subunits of a complex, so that the disulfide‐trapped complex can be displayed on the yeast surface for detection and analysis. Transient protein complexes are difficult to display on the yeast surface, since they may dissociate before they can be detected due to a long induction period in yeast. To this end, we show that three different quaternary structures with the subunit dissociation constant Kd ∼ 0.5–20 µM, the antibody variable domain (Fv), the IL‐8 dimer, and the p53–MDM2 complex, cannot be displayed on the yeast surface as a noncovalent complex. However, when we introduce an interchain disulfide between the subunits, all three systems are efficiently displayed on the yeast surface, showing that disulfide trapping can help display protein complexes that cannot be displayed otherwise. We also demonstrate that a disulfide forms only between the subunits that interact specifically, the displayed complexes exhibit functional characteristics that are expected of wt proteins, the mutations that decrease the affinity of subunit interaction also reduce the display efficiency, and most of the disulfide stabilized complexes are formed within the secretory pathway during export to the surface. Disulfide crosslinking is therefore a convenient way to study weak protein association in the context of yeast display. Biotechnol. Bioeng. 2010; 106: 27–41. © 2009 Wiley Periodicals, Inc.
High‐level rapid production of full‐size monoclonal antibodies in plants by a single‐vector DNA replicon system
Authors: Huang Zhong , Phoolcharoen Waranyoo , Lai Huafang , Piensook Khanrat , Cardineau Guy , Zeitlin Larry , Whaley Kevin J. , Arntzen Charles J. , Mason Hugh S. , Chen Qiang
Published: January 2010

Plant viral vectors have great potential in rapid production of important pharmaceutical proteins. However, high‐yield production of hetero‐oligomeric proteins that require the expression and assembly of two or more protein subunits often suffers problems due to the “competing” nature of viral vectors derived from the same virus. Previously we reported that a bean yellow dwarf virus (BeYDV)‐derived, three‐component DNA replicon system allows rapid production of single recombinant proteins in plants (Huang et al., 2009. Biotechnol Bioeng 103: 706–714). In this article, we report further development of this expression system for its application in high‐yield production of oligomeric protein complexes including monoclonal antibodies (mAbs) in plants. We showed that the BeYDV replicon system permits simultaneous efficient replication of two DNA replicons and thus, high‐level accumulation of two recombinant proteins in the same plant cell. We also demonstrated that a single vector that contains multiple replicon cassettes was as efficient as the three‐component system in driving the expression of two distinct proteins. Using either the non‐competing, three‐vector system or the multi‐replicon single vector, we produced both the heavy and light chain subunits of a protective IgG mAb 6D8 against Ebola virus GP1 (Wilson et al., 2000. Science 287: 1664–1666) at 0.5 mg of mAb per gram leaf fresh weight within 4 days post‐infiltration of Nicotiana benthamiana leaves. We further demonstrated that full‐size tetrameric IgG complex containing two heavy and two light chains was efficiently assembled and readily purified, and retained its functionality in specific binding to inactivated Ebola virus. Thus, our single‐vector replicon system provides high‐yield production capacity for hetero‐oligomeric proteins, yet eliminates the difficult task of identifying non‐competing virus and the need for co‐infection of multiple expression modules. The multi‐replicon vector represents a significant advance in transient expression technology for antibody production in plants. Biotechnol. Bioeng. 2010; 106: 9–17. © 2009 Wiley Periodicals, Inc.
Covalent dimerization of camelidae anti‐human TNF‐alpha single domain antibodies by the constant kappa light chain domain improves neutralizing activity
Authors: Giersberg Martin , Floss Doreen M. , Kipriyanov Sergey , Conrad Udo , Scheller Jürgen
Published: January 2010

The tumor necrosis factor‐alpha (TNFα) plays an important role in a number of chronic inflammatory disorders. Monoclonal camelidae variable heavy chain domain‐only antibodies (VHH) have been developed to antagonize the action of human TNFα (anti‐TNF‐VHH). Here we describe a strategy to obtain functional dimeric anti‐TNF‐VHH molecules, based on the C‐terminal fusion of a κ light chain domain to the anti‐TNF‐VHH. The resulting fusion protein was transiently expressed by use of viral vectors in Nicotiana benthamiana(Nb) leaves and purified. Competitive ELISA and cell cytotoxicity assays revealed that the dimerized anti‐NbTNF‐VHHCκ proteins blocked TNFα‐activity more effectively than either the monomeric Escherichia coli(Ec) produced anti‐EcTNF‐VHH or the monomeric anti‐NbTNF‐VHHCκ. We suggest that enhanced inhibition shown by dimeric anti‐NbTNF‐VHHCκ proteins is achieved by an increase in avidity. Biotechnol. Bioeng. 2010; 106: 161–166. © 2009 Wiley Periodicals, Inc.
Generation of a triple‐gene knockout mammalian cell line using engineered zinc‐finger nucleases
Authors: Liu Pei‐Qi , Chan Edmond M. , Cost Gregory J. , Zhang Lin , Wang Jianbin , Miller Jeffrey C. , Guschin Dmitry Y. , Reik Andreas , Holmes Michael C. , Mott John E. , Collingwood Trevor N. , Gregory Philip D.
Published: January 2010

Mammalian cells with multi‐gene knockouts could be of considerable utility in research, drug discovery, and cell‐based therapeutics. However, existing methods for targeted gene deletion require sequential rounds of homologous recombination and drug selection to isolate rare desired events—a process sufficiently laborious to limit application to individual loci. Here we present a solution to this problem. Firstly, we report the development of zinc‐finger nucleases (ZFNs) targeted to cleave three independent genes with known null phenotypes. Mammalian cells exposed to each ZFN pair in turn resulted in the generation of cell lines harboring single, double, and triple gene knockouts, that is, the successful disruption of two, four, and six alleles. All three biallelic knockout events were obtained at frequencies of >1% without the use of selection, displayed the expected knockout phenotype(s), and harbored DNA mutations centered at the ZFN binding sites. These data demonstrate the utility of ZFNs in multi‐locus genome engineering. Biotechnol. Bioeng. 2010; 106: 97–105. © 2009 Wiley Periodicals, Inc.
Multiple testing using the posterior probabilities of directional alternatives, with application to genomic studies
Authors: Labbe Aurélie , Thompson Mary E.
Published: January 2010

This paper aims to connect Bayesian analysis and frequentist theory in the context of multiple comparisons. The authors show that when testing the equality of two sample means, the posterior probability of the one‐sided alternative hypothesis, defined as a half‐space, shares with the frequentist P‐value the property of uniformity under the null hypothesis. Ultimately, the posterior probability may thus be used in the same spirit as a P‐value in the Benjamini‐Hochberg procedure, or in any of its extensions.
Bootstrapping data with multiple levels of variation
Authors: Field Christopher A. , Pang Zhen , Welsh Alan H.
Published: January 2010

The authors consider general estimators for the mean and variance parameters in the random effect model and in the transformation model for data with multiple levels of variation. They show that these estimators have different distributions under the two models unless all the variables have Gaussian distributions. They investigate the asymptotic properties of bootstrap procedures designed for the two models. They also report simulation results and illustrate the bootstraps using data on red spruce trees.
Generational risks for cancers not related to tobacco, screening, or treatment in the United States
Authors: Han Yueh‐Ying , Davis Devra L. , Weissfeld Joel L. , Dinse Gregg E.
Published: January 2010

BACKGROUND:

To assess trends in cancer, the authors evaluated the risk of 1 generation compared with that 25 years earlier (generational risk) for 3 groupings of cancers: those related to tobacco; those that reflect advances in screening or treatment; and a residual category of all other cancers.

METHODS:

In individuals ages 20 years to 84 years, age‐period‐cohort models were used to summarize time trends in terms of generational risk and average annual percentage change for US cancer incidence (1975‐2004) and mortality (1970‐2004) rates associated with these 3 cancer groupings.

RESULTS:

Adult white men today developed 16% fewer tobacco‐related cancers and had 21% fewer deaths because of those cancers than their fathers' generation, whereas adult white women experienced increases of 28% and 19%, respectively, relative to their mothers. The incidence of commonly screened cancers rose 74% in men and 10% in women, whereas mortality fell 25% in men and 31% in women. For cancers that have not been linked chiefly to tobacco or screening, the incidence was 34% and 23% higher in white men and white women, respectively, than in their parents' generation 25 years earlier. Mortality in this residual category decreased 14% in men and 18% in women. Results among blacks were qualitatively similar to those among whites.

CONCLUSIONS:

Despite declining overall cancer death rates, adults are experiencing increased incidence of cancers that are not associated with tobacco or screening relative to their parents. Future research should examine whether similar patterns are exhibited in other modern nations and should identify population‐wide avoidable risks that could account for unexplained increases in these residual cancers. Cancer 2010. © 2010 American Cancer Society.
Inducing Spin Crossover in Amphiphilic Iron(III) Complexes
Authors: Martinho Paulo N. , Harding Charles J. , Müller‐Bunz Helge , Albrecht Martin , Morgan Grace G.
Published: January 2010

Thermal spin crossover (SCO) was induced in a high‐spin FeIII complex by alkylation of the polyamino ligand backbone. The SCO profile was responsive to chain length with partial crossover observed with C6 alkylation and full transition upon lengthening to C12. No crossover was observed when the backbone was unfunctionalised, and all three complexes were obtained as unsolvated tetrafluoroborate salts.
Synthesis and Structural Characterization of Planar Chiral Cr(CO)3‐Complexed Aromatic Nitrones – Valuable Substrates for Asymmetric SmI2‐Induced Coupling Reactions
Authors: Chavarot‐Kerlidou Murielle , Rivard Michaël , Chamiot Bénédicte , Hahn Franck , Rose‐Munch Françoise , Rose Eric , Py Sandrine , Herson Patrick
Published: January 2010

The synthesis of Cr(CO)3‐complexed aromatic nitrones 2 is reported. These new planar‐chiral complexes were fully characterized in solution by 1H and 13C NMR, IR and cyclic voltammetry. Moreover, structural data were obtained from X‐ray structures of nitrones 2b, 2d, 2e and 2f. These analyses converge to give evidence of a favoured anti conformation of ortho‐substituted nitrones, with an unusual N–O···Haryl intramolecular interaction forming a six‐membered ring. The reactivity of these chiral nitrones in SmI2‐induced pinacol‐type reactions was investigated. The reductive cross‐coupling of nitrones 2a–d with carbonyl compounds proved to be highly chemo‐ and diastereoselective and afforded precursors of enantioenriched β‐amino alcohols in excellent yields.
Synthesis of 5‐Heteroaryl‐ and 5,7‐Bis(heteroaryl)azulenes by Electrophilic Substitution of 1,3‐Di‐tert‐Butylazulene with Triflates of N‐Containing Heterocycles
Authors: Shoji Taku , Ito Shunji , Toyota Kozo , Morita Noboru
Published: January 2010

An efficient synthesis of 5‐heteroaryl‐ and 5,7‐bis(heteroaryl)azulene derivatives was established for first time. The reaction proceeded through electrophilic substitution of 1,3‐di‐tert‐butylazulene (1) with the triflates of N‐containing heterocycles, in the presence of excess heterocycles, in good yields. The presumed intermediates, 5‐(dihydroheteroaryl)azulene derivatives were also available by the reaction with several N‐containing heterocycles in the presence of Tf2O under milder reaction conditions. Treatment of the 5‐(dihydroheteroaryl)azulene derivatives with KOH readily gave the desired 5‐(heteroaryl)azulene derivatives. Unexpectedly, N‐(5‐azulenyl)pyridinium triflate 17 was also obtained by the reaction of 1 with (trifluoromethylsulfonyl)pyridinium trifluoromethanesulfonate (TPT). 5‐Heteroaryl‐ and 5,7‐bis(heteroaryl)azulene derivatives exhibited a significant color change in acetic acid compared to those in dichloromethane due to the development of intramolecular charge‐transfer (CT) absorption bands. The redox behavior of these new azulene derivatives was examined by cyclic voltammetry (CV) and differential pulse voltammetry (DPV).
MVPACK: A package to calculate energy levels and magnetic properties of high nuclearity mixed valence clusters
Authors: Borrás‐Almenar J. J. , Cardona‐Serra S. , Clemente‐Juan J. M. , Coronado E. , Palii A. V. , Tsukerblat B.S.
Published: December 2009

We present a FORTRAN code based on a new powerful and efficient computational approach to solve the double exchange problem for high‐nuclearity MV clusters containing arbitrary number of localized spins and itinerant electrons. We also report some examples in order to show the possibilities of the program. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Report of a national conference on liver allocation in patients with hepatocellular carcinoma in the United States
Authors: Pomfret Elizabeth A. , Washburn Kenneth , Wald Christoph , Nalesnik Michael A. , Douglas David , Russo Mark , Roberts John , Reich David J. , Schwartz Myron E. , Mieles Luis , Lee Fred T. , Florman Sander , Yao Francis , Harper Ann , Edwards Erick , Freeman Richard , Lake John
Published: December 2009

A national conference was held to better characterize the long-term outcomes of liver transplantation (LT) for patients with hepatocellular carcinoma (HCC) and to assess whether it is justified to continue the policy of assigning increased priority for candidates with early-stage HCC on the transplant waiting list in the United States. The objectives of the conference were to address specific HCC issues as they relate to liver allocation, develop a standardized pathology report form for the assessment of the explanted liver, develop more specific imaging criteria for HCC designed to qualify LT candidates for automatic Model for End-Stage Liver Disease (MELD) exception points without the need for biopsy, and develop a standardized pretransplant imaging report form for the assessment of patients with liver lesions. At the completion of the meeting, there was agreement that the allocation policy should result in similar risks of removal from the waiting list and similar transplant rates for HCC and non-HCC candidates. In addition, the allocation policy should select HCC candidates so that there are similar posttransplant outcomes for HCC and non-HCC recipients. There was a general consensus for the development of a calculated continuous HCC priority score for ranking HCC candidates on the list that would incorporate the calculated MELD score, alpha-fetoprotein, tumor size, and rate of tumor growth. Only candidates with at least stage T2 tumors would receive additional HCC priority points. Liver Transpl 16:262–278, 2010. © 2009 AASLD.
Transcriptional regulation of pro‐apoptotic Par‐4 by NF‐κB/p65 and its function in controlling cell kinetics during early events in endometrial tumourigenesis
Authors: Saegusa Makoto , Hashimura Miki , Kuwata Takeshi , Okayasu Isao
Published: December 2009

Prostatic apoptosis response‐4 (Par‐4) was first identified in prostatic cancer cells that were induced to undergo apoptosis. Recently, Par‐4 has been suggested to be a tumour suppressor gene that plays a role in the development of endometrial carcinomas (ECs), but the exact mechanism remains to be clarified. Here we examined gene activation signalling cascades and influence on cell kinetics during endometrial tumourigenesis. In normal endometrium, constitutively high levels of Par‐4 expression were observed in epithelial cells through the menstrual cycle, in contrast to the transient up‐regulation in stromal components in the menstrual stage, correlated positively with the phospho‐p65 (pp65) status and apoptosis. In contrast, most ECs exhibited significant down‐regulation as compared to normal endometrium, with positive links only to pp65 expression. In EC cell lines, transfection of the NF‐κB subunit p65 led to transactivation of Par‐4 through specific binding to its promoter region, in contrast to the suppression by active Akt, suggesting that the balance between the two signals may be important to determine Par‐4 expression levels. In addition, transient overexpression of Par‐4 resulted in the induction of not only apoptosis but also senescence, through changes in the expression of bcl‐2 and p21$^{{\rm WAF1}}$, respectively. Together, these findings suggest that a signalling cascade involving sequential activation of NF‐κB/p65 and Par‐4 may participate in relatively early events of endometrial tumourigenesis, leading to modulation of cell kinetics including apoptosis and cell cycle progression. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Plasmin induces apoptosis of aortic valvular myofibroblasts
Authors: Kochtebane Najlah , Choqueux Christine , Passefort Stéphanie , Nataf Patrick , Messika‐Zeitoun David , Bartagi Aghleb , Michel Jean‐Baptiste , Anglés‐Cano Eduardo , Jacob Marie‐Paule
Published: December 2009

Previous studies have described remodelling of the extracellular substratum by matrix metalloproteinases (MMPs) in aortic valves. However, involvement of the fibrinolytic system has not yet been analysed. We hypothesized that plasminogen and plasminogen activator(s) are present in aortic valves and that plasminogen activation could induce the degradation of adhesive proteins and apoptosis of the valvular myofibroblasts. We employed ELISA, western blotting, fibrin‐agar zymography, and immunochemistry to detect components of the plasminogen activation system, in samples of aortic valves and valvular myofibroblasts in primary culture. Using myofibroblast cultures, real‐time measurement of plasminogen activation was performed in the absence and presence of inhibitors (amiloride, ε‐aminocaproic acid, and an MMP inhibitor); the degradation of fibronectin was visualized on western blots; and the apoptotic process was assessed by detection of phosphatidylserine exposure (binding of FITC‐annexin V) and DNA fragmentation (TUNEL and ELISA). We demonstrate that a time‐ and plasminogen concentration‐dependent generation of plasmin occurs on the surface of cultured valvular myofibroblasts expressing both u‐PA and t‐PA. Only u‐PA appears to activate plasminogen as t‐PA is essentially found in complex with PAI‐1. Plasmin‐dependent degradation of pericellular proteins, such as fibronectin, leads to cell detachment and apoptosis. In conclusion, various proteins of the fibrinolytic system are synthesized in vitro by cultured myofibroblasts from aortic valves, leading to plasmin‐dependent cell detachment‐induced apoptosis, a biological process named anoikis. The presence of plasminogen in aortic valves suggests that this process may be operating in vivo and may participate in valvular tissue remodelling, as also suggested by the finding of apoptotic cells in valvular tissue. This is the first demonstration of the presence and potential role of enzymes of the fibrinolytic system in aortic valves. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.