bluuebunny/crossref_metadata_2025 huggingface dataset:
Judicial Libraries in the Information Era: In the 21st Century, Role-Smart Law Librarians
Authors: Singh Karna , Maharana Bulu
Published: April 2024

The majority of legal studies take place in libraries. This paper discussed the services the judicial library helps legal practitioners render to members of the bar and bench. In the 21st century, information law plays a vital role and is the lifeblood of legal professionals. The "information age," also referred to as the "computer age," is defined by an explosion of information and fast technical innovation. This technological progress is the trend that is currently influencing legal research, teaching, and study. There are many social media platforms available today, but legal librarians in academic contexts face significant hurdles when users of their libraries find and use these technologies. In order to keep the library up-to-date and maintain a positive rapport with patrons, a librarian working in a library needs to possess a variety of skills, including curation, leadership, subject expertise in law, and research and technology abilities. The author outlines the competencies needed for a legal librarian in the digital age in this paper. This paper centers on them. On the issues raised, recommendations were made.
Standardisation of Centrifugation Method for Virgin Coconut Oil Production and Comparison of Quality Parameters of the Recovered Oil
Authors: Thanuja T. T. , Sreekala G. S.
Published: April 2024

Virgin coconut oil is a finest edible oil having immense health benefits. Several methods are adopted for the extraction of virgin coconut oil. This experiment aims to provide a cost-effective and efficient method for small-scale production of high-quality virgin coconut oil. The objective of this study was standardisation of centrifugation method for virgin coconut oil production. Mature coconuts from West Coast Tall variety were collected and coconut milk was taken out by using coconut gratings with equal quantity of water and coconut water and two fold of water.  The coconut milk was then chilled for 10 hours at 5-70C, followed by centrifugation at different revolutions per minute (rpm) and time. The colour, odour, taste, moisture content, free fatty acid content and total phenolic content of the resulting virgin coconut oil were assessed. The study revealed that the optimal conditions for maximum virgin coconut oil yield (28.087 percent) involve extracting coconut milk from coconut gratings with equal quantity of coconut water followed by chilling for 10 hours at 5-7ᵒC, centrifuged at 12000 rpm for 15 minutes and subsequently drying at 50°C.
A Study on the Psychological Impact of Covid-19 on Farmers
Authors: Vaishnavi C. , Kammar Shivanand K. , Goudappa S. B. , Wali Vijaya B. , B Prashanth.
Published: April 2024

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Site‐Specific Protein Labeling via Sortase‐Mediated Transpeptidation
Authors: Popp Maximilian Wei‐Lin , Antos John M. , Ploegh Hidde L.
Published: April 2009

Creation of functional protein bioconjugates demands methods for attaching a diverse array of probes to target proteins with high specificity, under mild conditions. The sortase A transpeptidase enzyme from Staphylococcus aureus catalyzes the cleavage of a short 5‐aa recognition sequence (LPXTG) with the concomitant formation of an amide linkage between an oligoglycine peptide and the target protein. By functionalizing the oligoglycine peptide, it is possible to incorporate reporters into target proteins in a site‐specific fashion. This reaction is applicable to proteins in solution and on the living cell surface. The method described in this unit only requires incubation of the target protein, which has been engineered to contain a sortase recognition site either at the C terminus or within solvent‐accessible loops, with a purified sortase enzyme and a suitably functionalized oligoglycine peptide. Curr. Protoc. Protein Sci. 56:15.3.1‐15.3.9. © 2009 by John Wiley & Sons, Inc.
Fall hazard control observed on residential construction sites
Authors: Kaskutas Vicki , Dale Ann Marie , Nolan James , Patterson Dennis , Lipscomb Hester J. , Evanoff Bradley
Published: April 2009

Background

Falls are a leading cause of mortality and morbidity in the construction industry. This study measured fall hazards at residential construction sites.

Methods

Trained carpenters administered the St. Louis Audit of Fall Risks and interviewed carpenters. The prevalence of fall prevention practices meeting safety criteria was counted and correlations explored.

Results

We identified a high prevalence of fall hazards at the 197 residential sites audited. Roof sheathing met safety criteria most consistently (81%) and truss setting least consistently (28%). Use of personal fall arrest and monitoring of unguarded floor openings were rare. Safer performance on several scales was correlated. Construction sites of large‐sized contractors were generally safer than smaller contractors. Apprentice carpenters were less familiar with their employers' fall prevention plan than experienced workers.

Conclusions

Safety could be improved with consistent use of recognized fall prevention practices at residential construction sites. Am. J. Ind. Med. 52:491–499, 2009. © 2009 Wiley‐Liss, Inc.
Mental retardation, facial anomalies, brachydactyly, cerebral angiomas, femoral nucleus necrosis: A new entity or Hall–Riggs syndrome?
Authors: Ficcadenti Anna , Santoro Lucia , Petroni Valeria , Carini Cecilia , Gabrielli Orazio
Published: April 2009

We report on a 4‐year‐old boy with mental retardation, facial and skeletal anomalies, cerebral angiomas, femoral nucleus necrosis, mild biochemical abnormalities. This complex of features resembles the Hall–Riggs syndrome but could represent a novel syndrome. © 2009 Wiley‐Liss, Inc.
Phenotype and 244k array‐CGH characterization of chromosome 13q deletions: An update of the phenotypic map of 13q21.1‐qter
Authors: Kirchhoff Maria , Bisgaard Anne‐Marie , Stoeva Radka , Dimitrov Boyan , Gillessen‐Kaesbach Gabriele , Fryns Jean‐Pierre , Rose Hanne , Grozdanova Liliana , Ivanov Ivan , Keymolen Kathelijn , Fagerberg Christina , Tranebjaerg Lisbeth , Skovby Flemming , Stefanova Margarita
Published: April 2009

Partial deletions of the long arm of chromosome 13 lead to variable phenotypes dependant on the size and position of the deleted region. In order to update the phenotypic map of chromosome 13q21.1‐qter deletions, we applied 244k Agilent oligonucleotide‐based array‐CGH to determine the exact breakpoints in 14 patients with partial deletions of this region. Subsequently, we linked the genotype to the patient's phenotype. Using this approach, we were able to refine the smallest deletion region linked to short stature (13q31.3: 89.5–91.6 Mb), microcephaly (13q33.3–q34), cortical development malformations (13q33.1‐qter), Dandy–Walker malformation (DWM) (13q32.2–q33.1), corpus callosum agenesis (CCA) (13q32.3–q33.1), meningocele/encephalocele (13q31.3‐qter), DWM, CCA, and neural tube defects (NTDs) taken together (13q32.3–q33.1), ano‐/microphthalmia (13q31.3–13qter), cleft lip/palate (13q31.3–13q33.1), lung hypoplasia (13q31.3–13q33.1), and thumb a‐/hypoplasia (13q31.3–q33.1 and 13q33.3–q34). Based on observations of this study and previous reports we suggest a new entity, “distal limb anomalies association,” linked to 13q31.3q33.1 segment. Most of the individuals with deletion of any part of 13q21qter showed surprisingly similar facial dysmorphic features, and thus, a “13q deletion facial appearance” was suggested. Prominent nasal columella was mapped between 13q31.3 and 13q33.3, and micrognathia between 13q21.33 and 13q31.1. The degree of mental delay did not display a particular phenotype–genotype correlation on chromosome 13. In contrast to previous reports of carriers of 13q32 band deletions as the most seriously affected patients, we present two such individuals with long‐term survival, 28 and 2.5 years. © 2009 Wiley‐Liss, Inc.
The genetics of twinning: From splitting eggs to breaking paradigms
Authors: Shur Natasha
Published: April 2009

Over the last decades, there has been a twin boom in terms of live births. Meanwhile, new insights into the genetics of twinning have changed major paradigms. The first major paradigm is that monozygotic (MZ) twins are identical. The second is that twins are either MZ or dizygotic (DZ). The third is that MZ twins happen by chance alone. We have discovered striking epigenetic and other differences between seemingly alike individuals; the presence of fascinating intermediate twin forms; and the potential for familial MZ twinning and even a twinning gene. As this article discusses, the aforementioned paradigm shifts have influenced novel research directions and improved clinical approaches to twin‐management. © 2009 Wiley‐Liss, Inc.
Familial monozygotic twinning: A report of seven pedigrees
Authors: Machin Geoffrey
Published: April 2009

Seven families contained 19 MZ twin pairs (2.7 pairs/family), diagnosed by low‐stringency variable number tandem repeats in DNA from placental tissue or blood. Chorion status was known in 10 pairs, 6 dichorionic, 4 monochorionic. Sex ratio was equal. Autosomal dominant inheritance is apparent. © 2009 Wiley‐Liss, Inc.
Non‐identical monozygotic twins, intermediate twin types, zygosity testing, and the non‐random nature of monozygotic twinning: A review
Authors: Machin Geoffrey
Published: April 2009

Monozygotic twins (MZ) are rarely absolutely “identical.” This review discusses the types of genetic/epigenetic and prenatal environmental post‐zygotic mechanisms that cause discordance within such twin pairs. Some of these mechanisms—ranging from heterokaryotypia to skewed X‐chromosome inactivation—may cause extreme discordance, but these extremes are merely the more emphatic examples of discordance that, to some degree, underlies the majority of MZ twin pairs. Because of the entrenched misconception that MZ twins are necessarily identical, many MZ twin pairs are mistakenly designated as dizygotic (DZ). Clinical benefits to accurate zygosity determination include correct solid organ transplantation matching, if one twin requires donation for a non‐genetically mediated disease; the opportunity of preventive management for disorders that do not manifest synchronously; and better counseling to parents regarding their individually unique, and often psychologically puzzling, twin offspring. In twin pairs with complex and confusing biological origins, more detailed zygosity testing may be required. For example, intermediate trigametic and tetragametic chimeric dizygotic twins are reviewed, some of whom are, nevertheless, monochorionic (MC). Because of inter‐fetal vascular anastomoses in MC twins, genetic results from blood samples may not accurately reflect discordance in solid organs. Previously, it was thought that MZ twinning was some sort of embryological fluke. However, familial monozygotic twinning is more common than suggested by the literature. Seven new families are presented in an accompanying paper. Despite the difficulties and dangers of twin pregnancy (especially so for MC twins), human twinning persists, and continues to both challenge and fascinate parents, clinicians and geneticists. © 2009 Wiley‐Liss, Inc.
A new dynamic 3D virtual methodology for teaching the mechanics of atrial septation as seen in the human heart
Authors: Schleich Jean‐Marc , Dillenseger Jean‐Louis , Houyel Lucile , Almange Claude , Anderson Robert H.
Published: April 2009

Learning embryology remains difficult, since it requires understanding of many complex phenomena. The temporal evolution of developmental events has classically been illustrated using cartoons, which create difficulty in linking spatial and temporal aspects, such correlation being the keystone of descriptive embryology. We synthesized the bibliographic data from recent studies of atrial septal development. On the basis of this synthesis, consensus on the stages of atrial septation as seen in the human heart has been reached by a group of experts in cardiac embryology and pediatric cardiology. This has permitted the preparation of three‐dimensional (3D) computer graphic objects for the anatomical components involved in the different stages of normal human atrial septation. We have provided a virtual guide to the process of normal atrial septation, the animation providing an appreciation of the temporal and morphologic events necessary to separate the systemic and pulmonary venous returns. We have shown that our animations of normal human atrial septation increase significantly the teaching of the complex developmental processes involved, and provide a new dynamic for the process of learning. Anat Sci Ed 2:69–77, 2009. © 2009 American Association of Anatomists.
Laparoscopy: Learning a new surgical anatomy?
Authors: Jiménez Angel Martin , Aguilar Jose‐Francisco Noguera
Published: April 2009

Operative laparoscopy has progressed rapidly in recent years, and this alternative to the conventional approach for abdominal surgery has allowed the description of new planes, spaces, and anatomic references as a result of the artificial rupture of the “anatomical continuum.” Magnified laparoscopic views and the ability to deeply explore anatomic features better demonstrate the basic anatomy. Therefore, even as laparoscopy requires a more profound knowledge of basic anatomy, it enhances our understanding of this anatomy. Current technology for recording and editing video‐taped sequences facilitates presentation on screen and can detail all aspects of interest, making such videos high value educational material for learners. Likewise, the experimental surgical laboratory is an indispensable setting for the development of these and other new technologies, transmission of their knowledge, and surgical progress. Anat Sci Ed 2:81–83, 2009. © 2009 American Association of Anatomists.
Web‐based interactive 3D visualization as a tool for improved anatomy learning
Authors: Petersson Helge , Sinkvist David , Wang Chunliang , Smedby Örjan
Published: April 2009

Despite a long tradition, conventional anatomy education based on dissection is declining. This study tested a new virtual reality (VR) technique for anatomy learning based on virtual contrast injection. The aim was to assess whether students value this new three‐dimensional (3D) visualization method as a learning tool and what value they gain from its use in reaching their anatomical learning objectives. Several 3D vascular VR models were created using an interactive segmentation tool based on the “virtual contrast injection” method. This method allows users, with relative ease, to convert computer tomography or magnetic resonance images into vivid 3D VR movies using the OsiriX software equipped with the CMIV CTA plug‐in. Once created using the segmentation tool, the image series were exported in Quick Time Virtual Reality (QTVR) format and integrated within a web framework of the Educational Virtual Anatomy (EVA) program. A total of nine QTVR movies were produced encompassing most of the major arteries of the body. These movies were supplemented with associated information, color keys, and notes. The results indicate that, in general, students' attitudes towards the EVA‐program were positive when compared with anatomy textbooks, but results were not the same with dissections. Additionally, knowledge tests suggest a potentially beneficial effect on learning. Anat Sci Ed 2:61–68, 2009. © 2009 American Association of Anatomists.
Online supportive interactions: Using a network approach to examine communication patterns within a psychosis social support group in Taiwan
Authors: Chang Hui‐Jung
Published: April 2009

A network approach was used to determine the overall supportive communication patterns constructed within the PTT psychosis support group in Taiwan, the largest bulletin board system in the Chinese‐speaking world. The full sequences of supportive interactions were observed over a $2{\textstyle{1 \over 2}}$‐year period from February 2004 to July 2006. The results indicated that the most exchanged support types were information and network links. All types of supportive communication networks were relatively sparse, yet small groups of cliques with different provision of support types formed within the psychosis group. Most of the online supportive interactions exchanged at dyadic and triadic levels. The overall supportive network was highly centralized. The overall findings with implications for future studies were discussed.
Highly avid magnetic bead capture: An efficient selection method for de novo protein engineering utilizing yeast surface display
Authors: Ackerman Margaret , Levary David , Tobon Gabriel , Hackel Benjamin , Orcutt Kelly Davis , Wittrup K. Dane
Published: April 2009

Protein engineering relies on the selective capture of members of a protein library with desired properties. Yeast surface display technology routinely enables as much as million‐fold improvements in binding affinity by alternating rounds of diversification and flow cytometry‐based selection. However, flow cytometry is not well suited for isolating de novo binding clones from naïve libraries due to limitations in the size of the population that can be analyzed, the minimum binding affinity of clones that can be reliably captured, the amount of target antigen required, and the likelihood of capturing artifactual binders to the reagents. Here, we demonstrate a method for capturing rare clones that maintains the advantages of yeast as the expression host, while avoiding the disadvantages of FACS in isolating de novo binders from naïve libraries. The multivalency of yeast surface display is intentionally coupled with multivalent target presentation on magnetic beads—allowing isolation of extremely weak binders from billions of non‐binding clones, and requiring far less target antigen for each selection, while minimizing the likelihood of isolating undesirable alternative solutions to the selective pressure. Multivalent surface selection allows 30,000‐fold enrichment and almost quantitative capture of micromolar binders in a single pass using less than one microgram of target antigen. We further validate the robust nature of this selection method by isolation of de novo binders against lysozyme as well as its utility in negative selections by isolating binders to streptavidin‐biotin that do not cross‐react to streptavidin alone. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009
Expression of antibodies using single‐open reading frame vector design and polyprotein processing from mammalian cells
Authors: Kunes Yune Z. , Gion Wendy R. , Fung Emma , Salfeld Jochen G. , Zhu Rong‐Rong , Sakorafas Paul , Carson Gerald R.
Published: April 2009

We describe a novel polyprotein precursor‐based approach to express antibodies from mammalian cells. Rather than expressing heavy and light chain proteins from separate expression units, the antibody heavy and light chains are contained in one single‐open reading frame (sORF) separated by an intein gene fused in frame. Inside mammalian cells this ORF is transcribed into a single mRNA, and translated into one polypeptide. The antibody heavy and light chains are separated posttranslationally, assembled into the functional antibody molecule, and secreted into culture medium. It is demonstrated that Pol I intein from P. horikoshii mediates protein splicing and cleavage reactions in mammalian cells, in the context of antibody heavy and light chain amino acid sequences. To allow the separation of antibody heavy chain, light chain, and the intein, we investigated a number of intein mutations designed to inhibit intein‐mediated splicing but preserve cleavage reactions. We have also designed constructs in which the signal peptide downstream from intein has altered hydrophobicity. The use of some of these mutant constructs resulted in more efficient antibody secretion, highlighting areas that can be further explored in improving such an expression system. An antibody secreted using one of the sORF constructs was characterized. This antibody has correct N‐terminal sequences for both of its heavy and light chains, correct heavy and light chain MW as well as intact MW as measured by mass spectrometry. Its affinity to antigen, as measured by surface plasmon resonance (SPR), is indistinguishable from that of the same antibody produced using conventional method. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009
Retrograde versus antegrade percutaneous aortic balloon valvuloplasty: Immediate, short‐ and long‐term outcome at 2 years
Authors: Cubeddu Roberto J. , Jneid Hani , Don Creighton W. , Witzke Christian F. , Cruz‐Gonzalez Ignacio , Gupta Rakesh , Rengifo‐Moreno Pablo , Maree Andrew O. , Inglessis Ignacio , Palacios Igor F.
Published: April 2009

Background:

The short‐ and long‐term vascular risks and hemodynamic benefits of antegrade versus retrograde percutaneous aortic balloon valvuloplasty (PAV) have not been clearly established. With the advent of percutaneous aortic valve replacement strategies, more valvuloplasties are being performed. The antegrade approach may reduce vascular complications, particularly in patients with peripheral vascular disease (PVD). Comparing the clinical efficacy and complications of each technique is warranted.

Methods:

A cohort of 157 consecutive patients undergoing PAV between 2000 and 2006 were included in the study. Of these, 46 (29%) patients underwent antegrade PAV and 111 (71%) retrograde PAV. Choice of vascular approach (antegrade or retrograde) were determined by operator preference. The rate of death, nonfatal vascular complications, and 2‐year survival was explored.

Results:

The mean age of the study population was 79 years. Patients undergoing antegrade PAV were more likely hypertensive (56% vs. 39%, P = 0.001) with PVD (41% vs. 18%, P = 0.004). Nevertheless, logistic Euroscores were no different between the groups (antegrade 18% vs. retrograde 14%; P = 0.30). Baseline and postprocedural valve areas were also similar. However, patients undergoing antegrade PAV had significantly fewer vascular complications (2% vs. 19%; P = 0.005). Two‐year follow‐up revealed no significant difference in death (antegrade 81% vs. retrograde 69%; P = 0.16), stroke, congestive heart failure, and surgical aortic valve replacement.

Conclusions:

The hemodynamic benefit of PAV occurs regardless of the selected vascular approach. The antegrade technique results in significantly fewer vascular complications and similar long‐term outcomes. Antegrade PAV is feasible and safe, particularly in patients with PVD. © 2009 Wiley‐Liss, Inc.
Microtubule involvement in the deposition of radial fibrillar callose arrays in stomata of the fern Asplenium nidus L.
Authors: Apostolakos Panagiotis , Livanos Pantelis , Galatis Basil
Published: April 2009

Aniline blue staining and callose immunolabeling revealed the deposition of significant callose quantities, in the form of fibrils, in the periclinal walls of guard cells (GCs) of stomata of the fern Asplenium nidus. The stomata that were at an early stage of differentiation displayed short callose fibrils at the junctions of the periclinal walls with the dorsal ones, which converged on the site of the future stomatal pore. In stomata being at an advanced stage of differentiation, callose fibrils were radially arranged around the stomatal pore, while in mature closed ones they were focused on the margins of the wall thickenings lining the stomatal pore. The pattern of the callose fibril organization resembled that of cellulose microfibrils in the same walls. Like the cellulose microfibrils, callose fibrils appeared coaligned with the underlying radial arrays of cortical microtubules (MTs). Moreover, the stomata treated with cellulose synthesis inhibitors (coumarin or dichlobenil) and those recovering from treatments with callose synthesis inhibitors (2‐deoxy‐D‐glucose or tunicamycin) exhibited distinct radial callose fibril arrays. Cytochalasin B did not affect the organization of the radial callose fibril arrays. In contrast, oryzalin completely disturbed the pattern of callose deposition in the affected GCs. Therefore, the fibrillar callose orientation in the periclinal GC walls is probably controlled by MTs but not by actin filaments. The MTs seem to orient callose synthases in the plasmalemma, thus determining the fibrillar nature of callose deposits and their radial mode of arrangement. The cellulose microfibrils are not involved in the callose fibril alignment. Cell Motil. Cytoskeleton 2009. © 2009 Wiley‐Liss, Inc.
The effects of extracellular calcium on motility, pseudopod and uropod formation, chemotaxis, and the cortical localization of myosin II in Dictyostelium discoideum
Authors: Lusche Daniel F. , Wessels Deborah , Soll David R.
Published: April 2009

Extracellular Ca++, a ubiquitous cation in the soluble environment of cells both free living and within the human body, regulates most aspects of amoeboid cell motility, including shape, uropod formation, pseudopod formation, velocity, and turning in Dictyostelium discoideum. Hence it affects the efficiency of both basic motile behavior and chemotaxis. Extracellular Ca++ is optimal at 10 mM. A gradient of the chemoattractant cAMP generated in the absence of added Ca++ only affects turning, but in combination with extracellular Ca++, enhances the effects of extracellular Ca++. Potassium, at 40 mM, can partially substitute for Ca++. Mg++, Mn++, Zn++, and Na+ cannot. Extracellular Ca++, or K+, also induce the cortical localization of myosin II in a polar fashion. The effects of Ca++, K+ or a cAMP gradient do not appear to be similarly mediated by an increase in the general pool of free cytosolic Ca++. These results suggest a model, in which each agent functioning through different signaling systems, converge to affect the cortical localization of myosin II, which in turn effects the behavioral changes leading to efficient cell motility and chemotaxis. Cell Motil. Cytoskeleton 2009. © 2009 Wiley‐Liss, Inc.
Distribution of P2X3‐immunoreactive fibers in hairy and glabrous skin of the rat
Authors: Taylor Anna M.W. , Peleshok Jennifer C. , Ribeiro‐da‐Silva Alfredo
Published: April 2009

The skin is innervated by two populations of unmyelinated sensory fibers, the peptidergic and nonpeptidergic, which transmit nociceptive information to the central nervous system. The peptidergic population expresses neuropeptides such as substance P (SP) and calcitonin gene‐related peptide (CGRP) and has both cutaneous and visceral targets. The nonpeptidergic population expresses the purinergic receptor P2X3, binds the isolectin B4 (IB4), and innervates mainly the epidermis. To date, the peptidergic nociceptor population in cutaneous tissue of the rat has been well characterized, whereas the nonpeptidergic innervation pattern has lacked an adequate description. To this aim, we used light microscopic immunocytochemistry to investigate the pattern of P2X3‐immunoreactive (‐IR) fiber innervation of both hairy and glabrous skin from male Sprague‐Dawley rats. Our results show extensive P2X3‐IR fibers throughout the upper and lower dermis. Thick bundles of P2X3‐IR fibers were found to run in parallel with the dermal‐epidermal junction and projected multiple thin collateral axons that penetrated the epidermal layer, creating a dense network of innervation throughout the entire epidermis. The distribution of P2X3‐IR fibers in the epidermis was far more extensive than the distribution of CGRP‐IR fibers. P2X3‐IR fibers also innervate hair follicles but were rarely found in close proximity to glands and blood vessels. The present results suggest a primary role for P2X3‐IR fibers in the detection of noxious stimuli in cutaneous tissue and provide an anatomical basis for future studies examining a possible functionally distinct role of nonpeptidergic nociceptors in the transmission of nociceptive signals. J. Comp. Neurol. 514:555–566, 2009. © 2009 Wiley‐Liss, Inc.
A tag‐less method of sorting stem cells from clinical specimens and separating mesenchymal from epithelial progenitor cells
Authors: Roda Barbara , Reschiglian Pierluigi , Zattoni Andrea , Alviano Francesco , Lanzoni Giacomo , Costa Roberta , Di Carlo Arianna , Marchionni Cosetta , Franchina Michele , Bonsi Laura , Bagnara Gian Paolo
Published: April 2009

Background:

The interest in stem cell (SC) isolation from easily accessible clinical specimens is booming. The lack of homogeneity in pluri/multipotent SC preparation blurs standardization, which however is recommended for successful applications. Multipotent mesenchymal SCs (MSCs) in fact express a broad panel of surface antigens, which limit the possibility of sorting homogeneous preparations by using an immunotag‐based method.

Methods:

We present a tag‐less, flow‐assisted method to purify, distinguish, and sort pluri/multipotent SCs obtained from clinical specimens, based on differences in the biophysical properties that cells acquire when in suspension under fluidic conditions. A suspension of cells in a transport fluid is injected into a ribbon‐like capillary device by continuous flow. In a relatively short time (about 30 min), sorted cells are collected.

Results:

We obtained baseline separation between MSCs and epithelial cells, which are important contaminants of isolated MSCs. The extent of separation is evaluated by flow cytometry through detection of a specific epithelial antigen. MSCs from various human sources also prove to have different, characteristic, highly‐reproducible fractionation profiles. Finally, we evaluated the dissimilar differentiation potential among cell fractions obtained from sorting a single MSC source. After differentiation induction, a fraction displayed a differentiation yield close to 100%, whereas unfractionated cells contained only 40% of responding cells.

Conclusions:

The results demonstrate that the method presented is able to obtain selected and well‐characterized living MSCs with an increased differentiation yield. Its reduced cost, full biocompatibility, and scale‐up potential could make this method an effective procedure for stem cell selection. © 2009 Clinical Cytometry Society
Genome‐wide association scans for secondary traits using case‐control samples
Authors: Monsees Genevieve M. , Tamimi Rulla M. , Kraft Peter
Published: April 2009

Genome‐wide association studies (GWAS) require considerable investment, so researchers often study multiple traits collected on the same set of subjects to maximize return. However, many GWAS have adopted a case‐control design; improperly accounting for case‐control ascertainment can lead to biased estimates of association between markers and secondary traits. We show that under the null hypothesis of no marker‐secondary trait association, naïve analyses that ignore ascertainment or stratify on case‐control status have proper Type I error rates except when both the marker and secondary trait are independently associated with disease risk. Under the alternative hypothesis, these methods are unbiased when the secondary trait is not associated with disease risk. We also show that inverse‐probability‐of‐sampling‐weighted (IPW) regression provides unbiased estimates of marker‐secondary trait association. We use simulation to quantify the Type I error, power and bias of naïve and IPW methods. IPW regression has appropriate Type I error in all situations we consider, but has lower power than naïve analyses. The bias for naïve analyses is small provided the marker is independent of disease risk. Considering the majority of tested markers in a GWAS are not associated with disease risk, naïve analyses provide valid tests of and nearly unbiased estimates of marker‐secondary trait association. Care must be taken when there is evidence that both the secondary trait and tested marker are associated with the primary disease, a situation we illustrate using an analysis of the relationship between a marker in FGFR2 and mammographic density in a breast cancer case‐control sample. Genet. Epidemiol. 33:717–728, 2009. © 2009 Wiley‐Liss, Inc.
Upregulation of manganese superoxide dismutase (SOD2) is a common pathway for neuroendocrine differentiation in prostate cancer cells
Authors: Quirós Isabel , Sáinz Rosa M. , Hevia David , García‐Suárez Olivia , Astudillo Aurora , Rivas Manuel , Mayo Juan C.
Published: April 2009

Despite improvements in diagnosis of advanced prostate cancer (PCa), treatment is not efficient and 5‐year survival is still low. Initially, the less abundant of cell types, neuroendocrine cells (NE), are involved in regulatory process but their physiological role is not fully understood. Among others, an increase in NE cells along with tumor progression has been commonly reported but their role in tumorigenesis or the molecular mechanisms of transdifferentiation is still a matter of debate. We have used human PCa cells (LNCaP) induced to differentiate to NE cells with several stimuli: androgen withdrawal, cyclic AMP or treatment with the antioxidant pineal hormone melatonin. PCa patients' specimens were also analyzed by western blotting and by immunocytochemistry. NE‐like LNCaP cells express high levels of mitochondrial superoxide dismutase (MnSOD/SOD2) in addition to NE markers. MnSOD upregulation is mediated by NFκB transcription factor, mainly through p65 translocation into the nuclei. More importantly, overexpression of MnSOD induces the rise of NE‐markers in LNCaP cells, showing that MnSOD upregulation might be instrumental for NE differentiation in PCa cells. Furthermore, MnSOD is highly expressed in advanced tumors of patients' when compared with control, nonpathological samples or with low‐grade tumors, along with the presence of synaptophysin, a common NE marker. Also, fluorescence immunohistochemical analysis revealed that MnSOD colocalizes with NE markers in most of NE cells observed in PCa specimens. The present findings indicate that MnSOD is essential for NE transdifferentiation and mediates in part the differentiation process, which appears also to be critical in vivo. © 2009 UICC
31P Solid‐State NMR study of the chemical setting process of a dual‐paste injectable brushite cements
Authors: Legrand A. P. , Sfihi H. , Lequeux N. , Lemaître J.
Published: April 2009

The composition and evolution of a brushite‐type calcium phosphate cement was investigated by Solid‐State NMR and X‐ray during the setting process. The cement is obtained by mixing β‐tricalcium phosphate [Ca3(PO4)2, β‐TCP] and monocalcium phosphate monohydrate [Ca(H2PO4)2.H2O, MCPM] in presence of water, with formation of dicalcium phosphate dihydrate or brushite [CaHPO2·2H2O, DCPD]. Analysis of the initial β‐TCP paste has shown the presence of β‐calcium pyrophosphate [Ca2P2O7, β‐CPy] and that of the initial MCPM a mixture of MCPM and dicalcium phosphate [CaHPO4, DCP]. Follow‐up of the chemical composition by 31P Solid‐State NMR enables to show that the chemical setting process appeared to reach an end after 20 min. The constant composition observed at the end of the process was similarly determined. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009
Solution of a conjecture of Tewes and Volkmann regarding extendable cycles in in‐tournaments
Authors: Meierling Dirk
Published: April 2009

A directed cycle C of a digraph D is extendable if there exists a directed cycle C′ in D that contains all vertices of C and an additional one. In 1989, Hendry defined a digraph D to be cycle extendable if it contains a directed cycle and every non‐Hamiltonian directed cycle of D is extendable. Furthermore, D is fully cycle extendable if it is cycle extendable and every vertex of D belongs to a directed cycle of length three. In 2001, Tewes and Volkmann extended these definitions in considering only directed cycles whose length exceed a certain bound 3≤k<n: a digraph D is k ‐extendable if every directed cycle of length t, where k≤t<n, is extendable. Moreover, D is called fully k ‐extendable if D is k ‐extendable and every vertex of D belongs to a directed cycle of length k. An in‐tournament is an oriented graph such that the in‐neighborhood of every vertex induces a tournament. This class of digraphs which generalizes the class of tournaments was introduced by Bang‐Jensen, Huang and Prisner in 1993. Tewes and Volkmann showed that every connected in‐tournament D of order n with minimum degree δ≥1 is ($n- \lfloor {{4\delta+1} \over {3}} \rfloor$) ‐extendable. Furthermore, if D is a strongly connected in‐tournament of order n with minimum degree δ=2 or $\delta {>}{{8n-17}\over {31}}$, then D is fully ($n-\lfloor {{4\delta+1} \over {3}}\rfloor$) ‐extendable. In this article we shall see that if$3 \leq \delta \leq {{8n-17} \over {31}}$, every vertex of D belongs to a directed cycle of length $n-\lfloor {{4\delta+1} \over {3}}\rfloor$, which means that D is fully ($n-\lfloor {{4\delta+1} \over {3}}\rfloor$) ‐extendable. This confirms a conjecture of Tewes and Volkmann. © 2009 Wiley Periodicals, Inc. J Graph Theory 63: 82–92, 2010
Synthesis and bioactivity evaluation of 3‐hydroxy‐3‐(phenylethynyl)indol‐2‐one analogues
Authors: Chen Gang , Wang Ye , Gao Suo , He Hong‐Ping , Li Shun‐Lin , Zhang Jian‐Xin , Ding Jian , Hao Xiao‐Jiang
Published: April 2009

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A series of propargylic alcohol was synthesized by the addition of phenylacetylene to isatin and its N‐substituted derivatives for the first time. This reaction involves activation of zinc reagent via coordination with carbonyl substrates that behave “ligand like.” The bioactivities on protective effect on the apoptosis of PC12 cells induced by H2O2 and cytotoxicity against lung cancer A549 and P388 cell line of these compounds were investigated, and several compounds showed potent activities. J. Heterocyclic Chem., 46, 217 (2009).
A palladium catalyzed new synthesis of N,N‐dimethyl[1,8]‐naphthyridine‐2‐amines: Facile incorporation of N,N‐dimethylamino group from DMF in aqueous medium
Authors: Goswami Shyamaprosad , Das Nirmal Kumar
Published: April 2009

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Here we report Pd catalyzed synthesis of N,N‐dimethyl[1,8]naphthyridine‐2‐amines by facile incorporation of N(Me)2 group from DMF in moderate to good yield. J. Heterocyclic Chem., 46, 324 (2009).
Microwave‐assisted synthesis, metallation, and duff formylation of porphyrins
Authors: Yaseen Muhammad , Ali Mukhtar , NajeebUllah Muhammad , Ali Munawar Munawar , Khokhar Irshad
Published: April 2009

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Microwave assisted synthesis, metallation, and β‐formylation of porphyrins is described. All synthetic reactions were carried out on inorganic polymer solid support using microwave energy. It is the first documented application of the Duff reaction in the field of porphyrins and metalloporphyrins. The overall process is simple, easy, and environment friendly. FTIR, UV–visible, elemental analysis, 1H NMR, and mass spectrometry were used to characterize the compounds. J. Heterocyclic Chem., 46, 251 (2009).
Gene expression during the oocyte‐to‐embryo transition in mammals
Authors: Evsikov Alexei V. , Marín de Evsikova Caralina
Published: April 2009

The seminal question in modern developmental biology is the origins of new life arising from the unification of sperm and egg. The roots of this question begin from 19th to 20th century embryologists studying fertilization and embryogenesis. Although the revolution of molecular biology has yielded significant insight into the complexity of this process, the overall orchestration of genes, molecules, and cells is still not fully formed. Early mammalian development, specifically the oocyte‐to‐embryo transition, is essentially under “maternal command” from factors deposited in the cytoplasm during oocyte growth, independent of de novo transcription from the nascent embryo. Many of the advances in understanding this developmental period occurred in tandem with application of new methods and techniques from molecular biology, from protein electrophoresis to sequencing and assemblies of whole genomes. From this bed of knowledge, it appears that precise control of mRNA translation is a key regulator coordinating the molecular and cellular events occurring during oocyte‐to‐embryo transition. Notably, oocyte transcriptomes share, yet retain some uniqueness, common genetic motifs among all chordates. The common genetic motifs typically define fundamental processes critical for cellular maintenance, whereas the unique genetic features may be a source of variation and a substrate for sexual selection, genetic drift, or gene flow. One purpose for this complex interplay among genes, proteins, and cells may allow for evolution to transform and act upon the underlying processes, at molecular, structural and organismal levels, to increase diversity, which is the ultimate goal of sexual reproduction. Mol. Reprod. Dev. 76: 805–818, 2009. © 2009 Wiley‐Liss, Inc.
Analytical solution to the transient phase of steady‐state free precession sequences
Authors: Ganter Carl
Published: April 2009

The transient phase of short–TR steady‐state free precession (SSFP) sequences exhibits an often striking complexity and is not only important for nonequilibrium applications (e.g., rapid T1–measurements), but can also cause severe artifacts in conventional imaging. In both cases, balanced SSFP sequences are practically (with regard to preparation efficiency) and conceptually (concerning the theoretical understanding of the decay) easier to handle their unbalanced counterparts, for which currently no theory is available. Based on a decomposition of coherence pathways into irreducible subpaths, an exact mathematical solution to the transient phase of unbalanced SSFP sequences is presented in this article, which also includes the known results for balanced SSFP and the steady state of arbitrary SSFP sequences as special cases. As an application, it is shown that the familiar Look–Locker expression for the accelerated magnetization recovery in RF‐spoiled sequences is only valid for T2 → 0. In addition to oscillatory perturbations, systematic deviations from the monoexponential decay are observed for T2 0 as a consequence of memory effects. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.
Consequence analysis to buildings from bursting cylindrical vessels
Authors: Ferradás Enrique González , Alonso Fernando Díaz , Pérez Juan Francisco Sánchez , Miñarro Marta Doval , Aznar Agustín Miñana , Gimeno José Ruiz
Published: April 2009

Damage effects to buildings must be taken into account when performing consequence analysis, because people indoors can be affected by such damage. The objective of this article is to provide a methodology for estimating damage to buildings from the pressure wave produced by bursting cylindrical vessels, by combining characteristic overpressure‐impulse‐distance curves with PROBIT equations. This methodology allows the damage to be shown in the same diagram as the overpressure, impulse, and distance. Diagrams and equations are presented to determine minor damage to brick buildings of four or less floors (broken windows, displacement of doors and window frames, tile displacement, etc), major structural damage (cracks in walls, collapse of some walls), and collapse (the damage is so extensive that the building is partially or totally demolished). © 2009 American Institute of Chemical Engineers Process Saf Prog, 2009
Atomic‐orbital‐symmetry based σ‐, π‐, and δ‐decomposition analysis of bond orders
Authors: Sizova Olga V. , Skripnikov Leonid V. , Sokolov Alexander Yu , Sizov Vladimir V.
Published: April 2009

The atomic‐orbital‐symmetry based (AOSB) scheme for the decomposition of Mayer and Wiberg bond orders into σ‐, π‐, and δ‐components is used to investigate different types of covalent bonds. Four series of compounds are studied: simple molecules with homonuclear bonds, inorganic molecules with polar heteronuclear bonds, [Ru(CN)5(XY)]q transition metal complexes with π‐acceptor ligands, and dimetal complexes with multiple metal–metal bonds. © 2009 Wiley Periodicals, Inc. Int J Quantum Chem, 2009
Nephelauxetic effect revisited
Authors: Tchougréeff Andrei L. , Dronskowski Richard
Published: April 2009

We readdress the well‐known nephelauxetic effect in coordination compounds of transition metal ions and analyze its possible origins presented in the literature. The initial hypothesis was to ascribe the reduction of the effective Racah parameters B and C of the electron–electron interaction in the complexes as compared to their respective free ion values (which is the essence of the nephelauxetic effect) to the expansion of d‐orbitals in the complex because of their quantum mechanical mixing with the orbitals of the ligands leading to delocalization. This picture necessarily leads to a rigid positive correlation between the amount of the d‐shell splitting controlled by the same delocalization and the amount of the renormalization of the interaction parameters. In fact, such a rigid relation does not exist and the so‐called spectrochemical and nephelauxetic series of the ligands composed according to the aforementioned amounts do not coincide in many points. An alternative explanation based not on the delocalization, but on polarization of the ligands had been proposed at the same time as the delocalization based one. Realistic estimates had been obtained on this basis, but the scheme had never been implemented on the atomic scale, which is necessary to enable renormalization of the aforementioned interaction parameters as a “built‐in” function of quantum chemical software. The required atomic resolution formulation of the polarization‐based model of the nephelauxetic effect is constructed in this work, in which a relation of such an approach to the general problematics of the “next generation” of semiempirical methods of quantum chemistry is also discussed. © 2009 Wiley Periodicals, Inc. Int J Quantum Chem, 2009
Theoretical mechanisms of the superoxide radical anion catalyzed by the copper‐zinc superoxide dismutase
Authors: Chen Dezhan , Wang Qingli , Zhang Honghong , Mi Shizhen , Wang Juliang , Zeng Qiang , Zhang Guiqiu
Published: April 2009

The disproportionation of superoxide radical anions catalyzed by copper‐zinc superoxide dismutase was investigated in detail using density functional theory. The structures of each stationary point and the transition states were located so that the reaction pathways were determined. The results indicate that the reactions proceed by two steps both for the oxidized process of superoxide radical anion and the reduced one. The rate for the determining step of the reaction (2) is in very good agreement with the experimental value. The solvation effect on the reaction was also discussed. © 2009 Wiley Periodicals, Inc. Int J Quantum Chem, 2010
Quantitative assessment of the effect of basis set superposition error on the electron density of molecular complexes by means of quantum molecular similarity measures
Authors: Salvador Pedro , Fradera Xavier , Duran Miquel
Published: April 2009

The Chemical Hamiltonian Approach (CHA) method is applied to obtain Basis Set Superposition Error (BSSE)‐free molecular orbitals at the Hartree–Fock (HF) and Density Functional Theory (DFT) levels of theory. To assess qualitatively the effect of the BSSE on the first‐order electron density, we had previously applied Bader's analysis of the intermolecular critical points located on the electron density, as well as density difference maps for several hydrogen bonded complexes. In this work, Quantum Molecular Similarity Measures are probed as an alternative avenue to properly quantify the electronic relaxation due to the BSSE removal by means of distance indices between the uncorrected and corrected charge densities. It is shown that BSSE contamination is more important at the DFT level of theory, and in some cases, changes on the topology of the electron density are observed upon BSSE correction. Inclusion of diffuse functions have been found to dramatically decrease the BSSE effect in both geometry and electron density. The CHA method represents a good compromise to obtain accurate results with small basis sets. © 2009 Wiley Periodicals, Inc. Int J Quantum Chem, 2009