bluuebunny/crossref_metadata_2025 huggingface dataset:
The heparan sulfate proteoglycan (HSPG) glypican‐3 mediates commitment of MC3T3‐E1 cells toward osteogenesis
Authors: Haupt Larisa M. , Murali Sadasivam , Mun Foong Kin , Teplyuk Nadiya , Mei Leong Fong , Stein Gary S. , van Wijnen Andre J. , Nurcombe Victor , Cool Simon M.
Published: May 2009

Heparan sulfate (HS) sugar chains attached to core proteoglycans (PGs) termed HSPGs mediate an extensive range of cell–extracellular matrix (ECM) and growth factor interactions based upon their sulfation patterns. When compared with non‐osteogenic (maintenance media) culture conditions, under established osteogenic culture conditions, MC3T3‐E1 cells characteristically increase their osteogenic gene expression profile and switch their dominant fibroblast growth factor receptor (FGFR) from FGFR1 (0.5‐fold decrease) to FGFR3 (1.5‐fold increase). The change in FGFR expression profile of the osteogenic‐committed cultures was reflected by their inability to sustain an FGF‐2 stimulus, but respond to BMP‐2 at day 14 of culture. The osteogenic cultures decreased their chondroitin and dermatan sulfate PGs (biglycan, decorin, and versican), but increased levels of the HS core protein gene expression, in particular glypican‐3. Commitment and progress through osteogenesis is accompanied by changes in FGFR expression, decreased GAG initiation but increased N‐ and O‐sulfation and reduced remodeling of the ECM (decreased heparanase expression) resulting in the production of homogenous (21 kDa) HS chain. With the HSPG glypican‐3 expression strongly upregulated in these processes, siRNA was used to knockdown this gene to examine the effect on osteogenic commitment. Reduced glypican‐3 abrogated the expression of Runx2, and thus differentiation. The reintroduction of this HSPG into Runx2‐null cells allowed osteogenesis to proceed. These results demonstrate the dependence of osteogenesis on specific HS chains, in particular those associated with glypican‐3. J. Cell. Physiol. 220: 780–791, 2009. © 2009 Wiley‐Liss, Inc.
Stepwise up‐regulation of MicroRNA expression levels from replicating to reversible and irreversible growth arrest states in WI‐38 human fibroblasts
Authors: Maes Olivier C. , Sarojini Harshini , Wang Eugenia
Published: May 2009

MicroRNAs (miRNAs) are small non‐coding RNAs that regulate diverse genetic expression networks through their control of mRNA stability or translation. Their role in aging mechanisms has been proposed in various model systems. In this report, the expression profiling of 462 human miRNAs in the reversible growth arrest state of quiescence, and irreversible states of replicative senescence and hydrogen peroxide‐induced premature senescence, are compared to young replicating lung fibroblasts. Greater numbers of up‐regulated than down‐regulated miRNAs are observed when cells stop proliferating, particularly in premature senescence, somewhat less in replicative senescence, and less still in quiescence. Several altered miRNA expressions are shared by the three growth arrest states, including the up‐regulation of miR‐34a, ‐624, ‐638 and miR‐377, and the down‐regulation of miR‐365 and miR‐512‐5p. miRNAs up‐regulated in both permanent growth arrest states but not in quiescence include let‐7g, miR‐26a, ‐136, ‐144, ‐195 and miR‐200b. In each of the growth arrest states, miR‐34a and let‐7f have the most robust up‐regulation in H2O2‐induced premature senescence, followed by miR‐638 and miR‐663 in replicative senescence, and finally, miR‐331‐3p and miR‐595 in quiescence. Our comprehensive evaluation of miRNA target correlations with known biomarkers for replicative senescence suggests that miRNAs may repress pathways controlling not only cell cycle traverse and proliferation, but also insulin‐like signaling, DNA repair and apoptosis, all of which are cellular functions deficient in senescent human fibroblasts. J. Cell. Physiol. 221: 109–119, 2009. © 2009 Wiley‐Liss, Inc
Osteopontin increases migration and MMP‐9 up‐regulation via αvβ3 integrin, FAK, ERK, and NF‐κB‐dependent pathway in human chondrosarcoma cells
Authors: Chen Ying‐Ju , Wei Ying‐Ying , Chen Hsien‐Te , Fong Yi‐Chin , Hsu Chin‐Jung , Tsai Chun‐Hao , Hsu Horng‐Chaung , Liu Shing‐Hwa , Tang Chih‐Hsin
Published: May 2009

Tumor malignancy is associated with several features such as proliferation ability and frequency of metastasis. Osteopontin (OPN), which abundantly expressed in bone matrix, is involved in cell adhesion, migration, invasion and proliferation via interaction with its receptor, that is, αvβ3 integrin. However, the effect of OPN on migration activity in human chondrosarcoma cells is mostly unknown. Here we found that OPN increased the migration and expression of matrix metalloproteinase (MMP)‐9 in human chondrosarcoma cells (JJ012 cells). RGD peptide, αvβ3 monoclonal antibody and MAPK kinase (MEK) inhibitors (PD98059 and U0126) but not RAD peptide inhibited the OPN‐induced increase of the migration and MMP‐9 up‐regulation of chondrosarcoma cells. OPN stimulation increased the phosphorylation of focal adhesion kinase (FAK), MEK and extracellular signal‐regulated kinase (ERK). In addition, treatment of JJ012 cells with NF‐κB inhibitor (PDTC) or IκB protease inhibitor (TPCK) inhibited OPN‐induced cell migration and MMP‐9 up‐regulation. Stimulation of JJ012 cells with OPN also induced IκB kinase α/β (IKK α/β) phosphorylation, IκBα phosphorylation, p65 Ser536 phosphorylation, and κB‐luciferase activity. The OPN‐mediated increases in MMP‐9 and κB‐luciferase activities were inhibited by RGD peptide, PD98059 or FAK and ERK2 mutant. Taken together, our results indicated that OPN enhances the migration of chondrosarcoma cells by increasing MMP‐9 expression through the αvβ3 integrin, FAK, MEK, ERK and NF‐κB signal transduction pathway. J. Cell. Physiol. 221: 98–108, 2009. © 2009 Wiley‐Liss, Inc
First‐trimester prenatal sonographic diagnosis of ectopia cordis in a twin gestation
Authors: Barbee Kristen , Wax Joseph R. , Pinette Michael G. , Cartin Angelina , Blackstone Jacquelyn
Published: May 2009

The 11–14‐week ultrasound examination allows early pregnancy dating, detection of major anomalies and multiple gestations, and accurate chorionicity determination. We describe a rare case of first‐trimester sonographic diagnosis of ectopia cordis in a dichorionic twin pregnancy, illustrating the benefits of early ultrasound in patient counseling and management. © 2009 Wiley Periodicals, Inc. J Clin Ultrasound, 2009
Diagnosis of short rib polydactyly syndrome type IV (Beemer–Langer syndrome) with cystic hygroma: A case report
Authors: Taori Kishor B. , Sharbidre Kedar G. , Krishnan Vijay , Kundargi Nischal , Kulkarni Bipin R. , Satkar Vinay , Bopche Sneha
Published: May 2009

Short rib polydactyly syndrome (SRPS) is a very rare congenital autosomal recessive inherited disease, classified into four subtypes. It has distinct imaging findings on prenatal sonography (US) and ancillary findings on both pre‐ and postnatal examinations may help classify individual cases into one of four subtypes. We report the US findings in a case of SRPS type IV (Beemer–Langer dysplasia) in a male fetus with multiple congenital anomalies, including cystic hygroma. The postnatal ultrasound, radiographic, and postmortem examinations helped to classify the SRPS as type IV. We believe this is the first documented case associating cystic hygroma and polydactyly. © 2009 Wiley Periodicals, Inc. J Clin Ultrasound 2009
Synthesis and in vitro study of a new class of methylene‐bis‐4,6‐diarylbenzo[d]isoxazoles as potential antifungal agents
Authors: Srinivas A. , Nagaraj A. , Sanjeeva Reddy Ch.
Published: May 2009

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A new class of methylene‐bis‐4,6‐diarylbenzo[d]isoxazoles 8a, 8b, 8c, 8d, 8e, 8f, 8g, 8h, 8i was synthesized by the reaction of methylene‐bis‐aryl‐6‐hydroxymethylene‐2‐cyclohexenone 6 with hydroxylamine hydrochloride, followed by aromatization with DDQ. Chemical structures of the newly synthesized compounds were elucidated by their IR, 1H NMR, 13C NMR, MS, and elemental analyses. Furthermore, all the compounds were screened for their antifungal activity against various fungi and compared with their monomeric compounds. Among the synthesized compounds, 8b, 8g, and 8i were found to be the most active against Candida albicans (ATCC 10231), Aspergillus fumigatus (HIC 6094), Trichophyton rubrum (IFO 9185), and Trichophyton mentagrophytes (IFO 40996). It is interesting to note that the compounds 8a, 8g, and 8i showed fungicidal activity toward C. albicans at the concentration of 3.12 μg/mL, which is less than the concentration of standard Amphotericin B. J. Heterocyclic Chem., 46, 497 (2009).
A facile and efficient synthesis of novel pyrimido[5,4‐b][4,7]phenanthroline‐9,11(7H,8H,10H,12H)‐dione derivatives via microwave‐assisted multicomponent reactions
Authors: Shi Feng , Yan Shu , Zhou Dianxiang , Tu Shujiang , Zou Xiang , Hao Wenjuan , Zhang Xiaohong , Han Zhengguo , Wu Shanshan , Cao Xudong
Published: May 2009

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A series of novel pyrimido[5,4‐b][4,7]phenanthroline‐9,11(7H,8H,10H,12H)‐dione derivatives were synthesized via microwave‐assisted three‐component reactions of barbituric acid, aldehyde, and quinolin‐6‐amine in DMF without catalyst. This facile synthesis not only offers an economical and efficient synthetic strategy to this class of significant compounds but also enriches the investigations on microwave‐assisted multicomponent reactions. Moreover, this protocol has the prominent advantages of short reaction time, good yields, low cost, easy operation, as well as environmental‐friendliness. J. Heterocyclic Chem., 46, 563 (2009).
The synthesis of 4‐(3,3‐dimethyl‐3H‐pyrrolo[2,3‐f]quinolin‐2‐yl)pyrazoles and 4‐(3,3‐dimethyl‐3H‐pyrrolo[3,2‐h]quinolin‐2‐yl)pyrazoles
Authors: Rashidi A. , Afghan A. , Baradarani Mehdi M. , Joule John A.
Published: May 2009

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5‐Hydrazinoquinoline and 8‐hydrazinoquinoline were converted via Fischer syntheses with 3‐methyl‐butan‐2‐one into pyrido‐indolenines 2,3,3‐trimethyl‐3H‐pyrrolo[2,3‐f]quinoline 7 and 2,3,3‐trimethyl‐3H‐pyrrolo[3,2‐h]quinoline 11, respectively. Exposure of the indolenines to the Vilsmeier reagent produced aminomethylene‐malondialdehydes 8 and 12, which reacted with hydrazine or arylhydrazines to give 4‐(3H‐pyrrolo[2,3‐f]quinolin‐2‐yl)‐ and 4‐(3H‐pyrrolo[3,2‐h]quinolin‐2‐yl)‐pyrazoles, 9 and 13. J. Heterocyclic Chem., 46, 428 (2009).
The alkylation of isatin‐derived oximes: Spectroscopic and X‐ray crystallographic structural characterization of oxime and nitrone products
Authors: Sin Ny , Venables Brian L. , Liu Xiaohong , Huang Stella , Gao Qi , Ng Alicia , Dalterio Richard , Rajamani Ramkumar , Meanwell Nicholas A.
Published: May 2009

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A series of isatin oximes was alkylated with alkyl halides and under Mitsunobu conditions to generate O‐alkylated oxime ether and N‐alkylated nitrone products. Alkylation of the sodium salts of oximes 5a and 5b with alkyl iodides produced predominantly the N‐alkylated nitrones (E)‐7 while alkylation of 5a and 5b with the harder electrophiles alkyl bromides and alkyl chlorides, gave mostly the O‐alkylated products, oxime ethers (E)‐6. Interestingly, alkylation of oxime 5b under Mitsunobu conditions with isopropyl alcohol produced the N‐alkylated nitrone (E)‐7bd as the major product. However, alkylation of oxime 5c, which incorporates a sterically bulky bromine substituent at the C‐4 position of the isatin heterocycle, with 2‐bromo‐ and 2‐iodo‐propane or with isopropyl alcohol under Mitsunobu conditions gave exclusively the O‐alkylated product, oxime ether (Z)‐6cd. The oxime ether and nitrone products (E)‐6 and (E)‐7, respectively, were characterized by LC/MS, 1H NMR, and 13C NMR. In addition, the structures of oxime ether (E)‐6bd and nitrone (E)‐7ad were determined by X‐ray crystallography. J. Heterocyclic Chem., 46, 432 (2009).
A facile electrochemical method for the synthesis of 5‐phenyl‐1,3,4‐oxadiazol‐2‐ylthio‐benzene‐1,2‐diol derivatives
Authors: Fakhari Ali Reza , Hosseiny Davarani Saied Saeed , Ahmar Hamid , Hasheminasab Kobra , Khavasi Hamid Reza
Published: May 2009

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Electrochemical oxidation of catechols to corresponding o‐quinones was successfully performed in aqueous solution by electrolysis at the controlled potentials. Quinones derived from catechols, participate in Michael addition reactions with 5‐phenyl‐1,3,4‐oxadiazole‐2‐thiol and via EC mechanism, converted to corresponding 5‐phenyl‐1,3,4‐oxadiazol‐2‐ylthio‐benzene‐1,2‐diol derivatives (4 and 4′). The products have been characterized using IR, 1H NMR, 13C NMR, X‐ray, and mass spectral data. J. Heterocyclic Chem., 46, 443 (2009).
Synthesis, structure, and some chemical properties of diferrocenyl‐1,2,3‐triazines
Authors: Klimova Elena I. , Klimova Tatiana , Álamo Marcos Flores , Iturbide Daniel Méndez , García Marcos Martínez
Published: May 2009

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The reactions of 1‐amino‐2,3‐diferrocenylcyclopropenylium tetrafluoroborate (5a, 5b, 5c) or 2,3‐diferrocenyl‐1‐methylthiocyclopropenylium iodide with sodium azide afford 5‐amino‐4,6‐diferrocenyl‐1,2,3‐triazines (7a, 7b, 7c) or 5‐methylthio‐4,6‐diferrocenyl‐ and 4‐methylthio‐5,6‐diferrocenyl‐1,2,3‐triazines (8a and 8b), respectively. Their structures were established using spectroscopic methods and that of compound 8a was confirmed using X‐ray diffraction analysis. Triazines 5a, 5b, 5c react with alkyl iodides to yield N(2)‐ and N‐alkyl‐C(5)‐derivatives. Their alkaline hydrolysis results in 2‐methyl‐4,6‐diferrocenyl‐2,5‐dihydro‐1,2,3‐triazin‐5‐one (16) and 4,6‐diferrocenyl‐5‐hydroxy‐1,2,3‐triazine (17). J. Heterocyclic Chem., 46, 477 (2009).
Insertion and fragmentation of 2‐ferrocenylmethylidene‐1, 3‐diketones upon their reactions with N‐methylhydrazine
Authors: Klimova Elena I. , Vázquez López Eduardo A. , Martínez Mendoza Juan M. , Ramírez Lena Ruíz , Alamo Marcos Flores , Backinowsky Leon V.
Published: May 2009

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Reactions of 2‐ferrocenylmethylidene‐1,3‐diketones (1a, 1b, 1c) with methylhydrazine afford mainly insertion products (∼40–58%), viz., 1‐(N′‐acyl‐N′‐methylhydrazino)‐1‐ferrocenyl‐2‐acylethanes (7a, 7b, 7c, 7d), together with lesser amounts of pyrazoles (8a, 8b) and dihydropyrazoles (9a, 9b). J. Heterocyclic Chem., 46, 484 (2009).
Stereoselective synthesis and biological activities of diethyl (E)‐{[4‐cyano‐5‐[[(disubstitutedamino)methylene]amino]‐3‐(methylthio)‐ 1H‐pyrazol‐1‐yl]substituited phenylmethyl}phosphonates
Authors: Xiao Lin‐Xia , Shi De‐Qing
Published: May 2009

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Diethyl {[5‐amino‐4‐cyano‐3‐(methylthio)‐1H‐pyrazol‐1‐yl]substitutedphenylmethyl}phosphonates 3 were efficiently synthesized via the condensation of [(1‐hydrazino)substitutedphenylmethyl]phosphonates 1 with 2‐[bis(methylthio)methylene]malononitrile 2. 3 reacted with triethyl orthoformate to afford diethyl (E)‐{[4‐cyano‐5‐[(ethoxymethylene)amino]‐3‐(methylthio)‐1H‐pyrazol‐1‐yl]substitutedphenylmethyl}phosphonates 4, which reacted with various secondary amines at room temperature to provide the target compounds 5 in good yields. Their structures were confirmed by ir, 1H and 31P NMR, mass spectroscopy, and elemental analyses. The results of preliminary bioassay indicated that compounds 5 possess potent herbicidal activity against the roots of dicotyledonous (oil rape) plants at the dosage of 100 mg/L, and compounds 5c and 5g exhibit 80.8% and 76.7% inhibitory activity against Colletotrichum gossypi at the concentration of 50 mg/L, respectively. Abstract end data: J. Heterocyclic Chem., 46, 555 (2009).
Patient readmissions, emergency visits, and adverse events after software‐assisted discharge from hospital: Cluster randomized trial
Authors: Graumlich James F. , Novotny Nancy L. , Stephen Nace G. , Kaushal Himangi , Ibrahim‐Ali Waleed , Theivanayagam Shoba , William Scheibel L. , Aldag Jean C.
Published: May 2009

BACKGROUND:

One of the causes of postdischarge adverse events is poor discharge communication between hospital‐based physicians, patients, and outpatient physicians. The value of hospital discharge software to improve communication and clinically relevant outcomes is unknown.

OBJECTIVE:

To measure effects of a discharge software application of computerized physician order entry (CPOE).

DESIGN:

Cluster randomized controlled trial.

SETTING:

Tertiary care, teaching hospital in central Illinois.

PATIENTS:

A total of 631 inpatients discharged to home with high risk for readmission.

INTERVENTION:

Seventy internal medicine hospital physicians were randomly assigned (allocation concealed) to discharge software versus usual care, handwritten discharge.

MEASUREMENTS:

Blinded assessment of patient readmission, emergency department visit, and postdischarge adverse event.

RESULTS:

A total of 590 (94%) patients provided 6‐month follow‐up data. Generalized estimating equations gave intervention variable coefficients with 95% confidence interval (CI). When comparing patients assigned to discharge software versus usual care, there was no difference in hospital readmission within 6 months (37.0% versus 37.8%; coefficient −0.005 [95% CI, −0.074 to 0.065]; P = 0.894), emergency department visit within 6 months (35.4% versus 40.6%; coefficient −0.052 [95% CI, −0.115 to 0.011]; P = 0.108), or adverse event within 1 month (7.3% versus 7.3%; coefficient 0.003 [95% CI; −0.037 to 0.043]; P = 0.884).

CONCLUSIONS:

Discharge software with CPOE did not affect readmissions, emergency department visits, or adverse events after discharge. Future studies should assess other endpoints such as patient perceptions or physician perceptions to see if discharge software has value. Journal of Hospital Medicine 2009;4:E11–E19. © 2009 Society of Hospital Medicine.
Pushing lenders to over‐comply with environmental regulations: A developing country perspective
Authors: Kulkarni Parashar
Published: May 2009

In the past few years, the number of lenders adopting voluntary environmental codes, such as the Equator Principles, is increasing. The main reasons for this form of over‐compliance include warm glow preferences of agents, credit risk and incentives arising from regulation. Empirical evidence suggests that lenders that over‐comply are generally bigger than those that don't. In particular, they are likely to be MNCs. In addition, the behaviour of over‐complying lenders differs from other lenders, for instance they are more likely to incorporate environmental risks in their lending practices. In the context of developing countries, incentives that promote over‐compliance exist to much lesser degree. Analysing the regulatory environment in developing countries, we find no compelling reason for regulators to encourage voluntary initiatives such as the Equator Principles. Copyright © 2009 John Wiley & Sons, Ltd.
The impact of the Asian financial crisis on bank efficiency: The 1997 experience of Malaysia and Thailand
Authors: Sufian Fadzlan
Published: May 2009

In the mid‐1990s, the East Asian countries experienced severe financial crisis that were followed by deep economic downturns. A variety of methodologies have been used to understand the nature of the Asian financial crisis. However, the impact of the 1997 Asian financial crisis on the efficiency of the financial industry has yet to be studied. By employing the Data Envelopment Analysis (DEA) approach the present study attempts to examine for the first time the impact of the 1997 Asian financial crisis on the efficiency of the Malaysian and Thailand banking sectors, two of the East Asian countries that were severely affected by the crisis. The study focuses on three major approaches vis., intermediation, value added and operating approaches. The results clearly bring forth the high degree of inefficiency in the Malaysian and Thailand banking sectors, particularly a year after the crisis. We find that the Malaysian banking sector has exhibited a higher TE levels during the post crisis period under the intermediation and value added approaches, while TE seems to be lower under the operating approach. The empirical findings suggest that the Thailand banking sector has exhibited a lower TE level during the post crisis period under all approaches. Copyright © 2009 John Wiley & Sons, Ltd.
Design and synthesis of sulfur‐35 agents and their applications for protein labeling
Authors: Ren Sumei , McNamara Paul , Koharski David , Hesk David , Borges Scott
Published: May 2009

Two new 35S reagents were developed to radiolabel proteins. The first reagent, N‐succinimidyl‐4‐(methane [35S]sulfonylamino‐methyl)‐benzoate (SMSB), acylates the ε‐amino group of lysine residues in proteins. The second reagent, 4‐(methane [35S]sulfonylamino‐methyl)‐phenylpropylaldehyde (MSAPPA), labels lysine residues via reductive alkylation. Comparing the two methods, the reductive alkylation method labeled proteins over a broader pH range with higher overall radiochemical yield. More than ten monoclonal antibodies (mAbs) have been labeled with these 35S labeling reagents, the biological activity of the mAbs was unchanged. Part of this work was presented in the Ninth International Symposium on the Synthesis and Applications of Isotopically Labelled Compounds, Edinburgh, 16–20 July 2006. Copyright © 2009 John Wiley & Sons, Ltd.
A comparison of American and international prototypes of successful managers
Authors: Lankau Melenie J. , Chung Beth G.
Published: May 2009

In the present study, similarities and differences between prototypes of successful managers were examined across four cultural groups: Americans, Europeans, Asians, and Latin Americans. Managers from the hospitality industry (N = 366) used an 84–item attribute inventory to rate a successful middle manager. In addition, Americans' stereotypes of ethnic managers were compared with prototypes held by managers from those ethnic cultures. Specifically, American managers' perceptions of Asian and Hispanic managers were compared against Asian and Hispanic/Latin American managers' prototypes. A high level of correspondence in prototype characteristics was found across the four cultural groups. In addition, American‐defined ethnic manager stereotypes also contained profiles similar to cultural prototypes. However, important differences were also detected on many managerial characteristics. Implications of the findings for cross‐cultural congruence and areas for future research are discussed.
Intercultural and sustainable leadership: A professional program for emerging educational leaders
Authors: Frawley Jack
Published: May 2009

This research reports on an investigation into the changing perspectives of educational leaders from the Asia‐Pacific region attending an AusAID‐funded Australian Leadership Award Fellowships (ALAF) program hosted by Australian Catholic University's (ACU National) Flagship for Creative and Authentic Leadership (the Flagship). The ALAF program aimed to strengthen the capacity of educational leaders from the Asia‐Pacific region to work more effectively in education systems and to better manage the competing demands of governments, education bureaucracies, educational organizations, and funding agencies. The research focused on how the Fellows viewed leadership, and how they viewed themselves as leaders before and at the conclusion of the program. What emerged was a growing interest in intercultural leadership and the concern for sustainable leadership.
Gape and bite force in the rodents Onychomys leucogaster and Peromyscus maniculatus: Does jaw‐muscle anatomy predict performance?
Authors: Williams Susan H. , Peiffer Erika , Ford Sonya
Published: May 2009

Compared with the deer mouse, Peromyscus maniculatus, the grasshopper mouse, Onychomys leucogaster, exhibits modifications in its jaw‐muscle architecture that promote wide gapes and large bite forces at wide gapes to prey upon large vertebrate prey. In this study, we determine whether jaw‐muscle anatomy predicts gape and biting performance in O. leucogaster, and we also assess the influence of gape on bite force in the two species. Although O. leucogaster has an absolutely longer jaw, which facilitates larger gapes, maximum passive gape is similar in both species, averaging ∼12.5 mm. Thus, when scaled to jaw length, O. leucogaster has a smaller maximum passive gape. These results suggest that predatory behaviors of O. leucogaster may not require remarkably large gapes. On the other hand, both absolute and relative bite forces exerted by O. leucogaster are significantly larger than those of P. maniculatus. The largest bite forces in both species occur at 5.0 mm of gape at the incisors, or 40% of maximum gape. Although bite force in both species decreases at larger gapes, O. leucogaster does maintain a larger percentage of maximum bite force at gapes larger than 40% of maximum passive gape. Therefore, although structural modifications in the masticatory apparatus of O. leucogaster may constrain gape, they may help to maintain bite force at large gapes. These results suggest that increases in gape differentially influence the length‐tension properties of the jaw muscles in the two species. Finally, these results highlight the importance of considering the effect of muscle stretch on force production in comparative studies of bite force. As a first approximation, it appears that gapes of 40–50% of maximum gape in rodents optimizes bite force production at the incisors. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.
Combinatorial de novo design and application of a biomimetic affinity ligand for the purification of human anti‐HIV mAb 4E10 from transgenic tobacco
Authors: Platis Dimitris , Maltezos Anastasios , Ma Julian K‐C. , Labrou Nikolaos E.
Published: May 2009

Monoclonal anti‐HIV antibody 4E10 (mAb 4E10) is one of the most broadly neutralizing antibodies against HIV, directed against a specific epitope on envelope protein gp41. In the present study, a combinatorial de novo design approach was used for the development of a biomimetic ligand for the affinity purification of mAb 4E10 from tobacco transgenic extract in a single chromatographic step. The biomimetic ligand (4E10lig) was based on a L‐Phe/β‐Ala bi‐substituted 1,3,5‐triazine (Trz) scaffold (β‐Ala‐Trz‐L‐Phe, 4E10lig) which potentially mimics the more pronounced electrostatic and hydrophobic interactions of mAb 4E10‐binding sequence determined by screening of a random peptide library. This library was comprised of Escherichia coli cells harboring a plasmid (pFlitrx) engineered to express a fusion protein containing random dodecapeptides that were inserted into the active loop of thioredoxin, which itself was inserted into the dispensable region of the flagellin gene. Adsorption equilibrium studies with this biomimetic ligand and mAb 4E10 determined a dissociation constant (KD) of 0.41 ± 0.05 µM. Molecular modeling studies of the biomimetic ligand revealed that it can potentially occupy the same binding site as the natural binding core peptide epitope. The biomimetic affinity adsorbent was exploited in the development of a facile mAb 4E10 purification protocol, affording mAb 4E10 of high purity (approximately 95%) with good overall yield (60–80%). Analysis of the antibody preparation by SDS‐PAGE, enzyme‐linked immunosorbent assays (ELISA), and western blot showed that the mAb 4E10 was fully active and free of degraded variants, polyphenols, and alkaloids. Copyright © 2009 John Wiley & Sons, Ltd.
High prevalence of human papillomavirus type 58 in patients with cervical pre‐malignant lesions in southern Brazil
Authors: Paesi Suelen , Serafini Eduardo Pretto , Barea Fernanda , Madi Sonia Regina Cabral , Echeverrigaray Sergio
Published: May 2009

Cervical cancer is the second most common type of cancer in women worldwide. Several human papillomavirus (HPV) genotypes, sexual behavior, and socioeconomic profile represent major risk factors for the development of this carcinoma. Cervical invasive cancer is preceded by cellular abnormalities that can be identified by cytological or histological exams. In order to determine the prevalence and genotypes of HPV in women with abnormal cytology or histopathology, cervical cell samples from 256 patients were evaluated for the presence of HPV/DNA by polymerase chain reaction (PCR), followed by virus genotyping by restriction fragment length polymorphism (RFLP). A total of 113 samples (51.2%) were HPV/DNA positive. Viral genotyping showed that the most prevalent genotypes were HPV 16 (34.7%) and 58 (13.8%), followed by HPV 33 (9.72%), 11 (8.33%), 18 (5.55%), 53 (5.55%), and 6 (4.2%). Four samples (5.55%) exhibited multiple infections due to the great similarity of socioeconomic characteristics and sexual behavior of HPV positive women, it was not possible to establish a risk profile for female HPV infection. J. Med. Virol. 81:1270–1275, 2009. © 2009 Wiley‐Liss, Inc.
Epidemic spread of hepatitis C virus genotype 3a and relation to high incidence of hepatocellular carcinoma in Pakistan
Authors: Khan Anis , Tanaka Yasuhito , Azam Zahid , Abbas Zaigham , Kurbanov Fuat , Saleem Uzma , Hamid Saeed , Jafri Wasim , Mizokami Masashi
Published: May 2009

Studies conducted in different populations worldwide revealed an association between HCV genotype 1 and the development of hepatocellular carcinoma (HCC) than in infection with other HCV genotypes. There are reports which reveal the association of HCV genotype 3a (HCV‐3a) with hepatic steatosis and fibrosis but its relation with the development of HCC has not been investigated. In Pakistan, where the incidence of HCC is increasing, 189 patients with chronic liver disease including 82 with HCC were enrolled. HCV genotypes were determined by phylogeny in the NS5B region and the epidemic history of HCV‐3a was examined using coalescent theory based methods. HCV‐3a was the predominant genotype (81.4%) in the cohort studied, followed by 3b (9.3%), 3k (2.3%), 1a (1.5%), 1c (1.5%), 1b (0.8%), and 2a (0.8%) where 76% of HCC and 86% of non‐HCC were infected with HCV‐3a. The significant factors associated with HCC were older age (mean ± SD) 55.8 (±9.9) (P < 0.0001), and male gender (P < 0.001). HCV RNA was significantly higher in patients with HCC and chronic hepatitis than in liver cirrhosis (P < 0.0001). Molecular evolutionary analysis revealed a distinct phylogenetic cluster of HCV‐3a in Pakistan and an estimation of the effective number of HCV infections indicated the appearance of HCV‐3a in this region around 1920s and a rapid exponential growth in the 1950s. This indicates that the epidemic spread of HCV‐3a occurred earlier in Pakistan than in other countries in which this genotype has been reported. HCV‐3a which spread earlier in Pakistan may be associated with an increasing incidence of HCC. J. Med. Virol. 81:1189–1197, 2009. © 2009 Wiley‐Liss, Inc.
Hepatitis C virus infection in the family setting of patients with occult hepatitis C
Authors: Castillo Inmaculada , Bartolomé Javier , Quiroga Juan Antonio , Barril Guillermina , Carreño Vicente
Published: May 2009

Family members of patients with chronic hepatitis C virus (HCV) infection are at increased risk of HCV infection but the prevalence of HCV among family members of patients with occult HCV infection is not known. Anti‐HCV, serum HCV RNA and levels of liver enzymes were determined in 102 family members of 50 index patients with occult HCV infection and in 118 family members of 59 chronic hepatitis C index patients. HCV RNA and/or anti‐HCV were detected in 10/102 (9.8%) relatives of patients with occult HCV infection and in 4/118 (3.4%) of patients with chronic hepatitis C. Fourteen additional family members (seven were relatives of index patients with occult HCV infection) had abnormal values of liver enzymes without serological markers of HCV infection. Two of these patients (who were relatives of two index patients with occult HCV infection) underwent a liver biopsy and were diagnosed with an occult HCV infection because HCV RNA was detected in the liver cells in the absence of serological HCV markers. In conclusion, the prevalence of HCV infection among family members of patients with occult HCV infection was similar to that found among family members of patients with chronic hepatitis C. This stresses the need to adopt strategies to prevent the transmission of HCV in the family setting of patients with occult HCV infection. J. Med. Virol. 81:1198–1203, 2009. © 2009 Wiley‐Liss, Inc.
TLR3 gene polymorphisms and liver disease manifestations in chronic hepatitis C
Authors: Askar Eva , Bregadze Rusudan , Mertens Jasmin , Schweyer Stefan , Rosenberger Albert , Ramadori Giuliano , Mihm Sabine
Published: May 2009

Phenotypes of liver disease due to chronic hepatitis C virus (HCV) infection show a wide range of variations in terms of histological manifestations and the clinical outcome. Sensing of viral double‐stranded RNA (dsRNA) by Toll‐like receptor 3 (TLR3) is likely involved in early pathogen detection and the host response to viral infection. This study analyzed epidemiological and clinical data from a total of 137 patients with chronic HCV infection with regard to two polymorphic positions within the TLR3 gene: rs5743305 (T/A) is located within the promoter region and might affect transcriptional activity, rs3775291 (C/T) is a non‐synonymous single nucleotide polymorphism (SNP) located within exon 4 and the variant receptor has been shown to be functionally impaired. TLR3 promoter and the exon 4 variations were not found to be associated with TLR3 gene expression in peripheral blood mononuclear cells (PBMCs). In the liver, however, a tendency of higher TLR3 gene expression was found for exon 4 TT genotypes. Both variations were not found to be associated with clinical parameters of chronic disease. On the other hand, an analysis of the TLR3 exon 4 genotype distribution with respect to HCV subtype revealed an absence of TT genotype among HCV subtype 1a infected individuals. This study thus failed to reveal any association of the two SNPs under investigation with clinical parameters of chronic hepatitis C. However, data argue for a functional relevance of the exon 4 SNP in terms of conferring a different susceptibility towards HCV subtype infection. J. Med. Virol. 81:1204–1211, 2009. © 2009 Wiley‐Liss, Inc.
Investigation of pre‐diagnostic virological markers for progressive multifocal leukoencephalopathy in human immunodeficiency virus‐infected patients
Authors: Grabowski Mary K. , Viscidi Raphael P. , Margolick Joseph B. , Jacobson Lisa P. , Shah Keerti V.
Published: May 2009

Progressive multifocal leukoencephalopathy (PML) is a severe neurological disorder due to JC virus (JCV) infection. Pre‐diagnostic biological markers and risk factors for PML are not well understood. We conducted a case–control study nested within the Multicenter AIDS Cohort Study to examine the association between JCV viruria and viremia and serum antibody to JCV capsids, in relation to subsequent PML diagnoses, 5 months to 12 years later. Other demographic and immunologic factors were also examined. The study population included 28 incident cases of PML, 26 matched HIV‐positive controls, and 50 HIV‐negative controls. Prevalence of JCV viruria was 37% in cases, 42% in HIV‐positive controls, and 28% in HIV‐negative controls (P = 0.43). Among persons with JCV viruria, persistent viruria was more common in cases (89%) than in HIV‐positive controls (33%) (P = 0.02). Presence of JCV viruria was not related to the time to PML diagnosis (OR: 1.03, 95% CI: 0.8–1.4); however, the urinary concentration of JCV DNA increased with proximity to the date of PML diagnosis in cases. JCV seropositivity did not differ between cases or controls (P = 0.42). Four cases tested JCV seronegative, including one case only 5 months prior to diagnosis with PML. JCV DNA was detected in the serum of one HIV‐positive control. Smoking was the only demographic variable analyzed associated with an increased risk for PML (MOR: 9.0, 95% CI: 1.2–394.5). The results suggest that persistent JCV viruria and increasing urinary concentration of JCV DNA may be predictive of PML for some patients. J. Med. Virol. 81:1140–1150, 2009. © 2009 Wiley‐Liss, Inc.
Molecular analysis of HPV 16 E6I/E6II spliced mRNAs and correlation with the viral physical state and the grade of the cervical lesion
Authors: Cricca Monica , Venturoli Simona , Leo Elisa , Costa Silvano , Musiani Monica , Zerbini Marialuisa
Published: May 2009

The presence of HPV 16 E6*I/E6*II spliced transcripts, in cervical lesions of different grade, was analyzed to characterize the transcription pattern. The presence and amount of spliced transcripts were correlated with DNA viral markers such as E2/E6 ratio and physical state. The detection of HPV 16 E6*I/E6*II mRNAs was set up by an SYBR Green real‐time reverse transcriptase PCR assay with an optimal dynamic range and sensitivity. The assay was applied to the analysis of 71 specimens, positive to HPV 16 as a sole infection, from women with abnormal cervical smears, precisely 31 low‐grade squamous intraepithelial lesions and 40 high‐grade lesions. Samples negative to both transcripts were found only in low‐grade cervical lesions. Three different transcription profiles were found in the low‐ and high‐grade lesions analyzed: in low‐grade lesions samples positive only to E6*II and in high‐grade lesions samples positive only to E6*I were detected. In low‐ and high‐grade lesions, samples positive to both E6*I and E6*II were found. In the samples positive for both transcripts, the E6*I/E6*II ratio was higher than that in the majority of high‐grade lesions and lower than that in all the low‐grade lesions. Analyzing the transcription pattern in relation to E2/E6 ratio and to the DNA physical state, the presence of high values of E6*I was associated mainly with low values of E2/E6 ratio and of mixed DNA forms. The detection of HPV 16 E6*I/E6*II mRNAs may serve to identify transcription patterns indicative of cervical disease progression and help physicians to decide clinical management. J. Med. Virol. 81:1276–1282, 2009. © 2009 Wiley‐Liss, Inc.
Enterovirus 71‐induced autophagy detected in vitro and in vivo promotes viral replication
Authors: Huang Shu‐Chen , Chang Chia‐Lun , Wang Po‐Shun , Tsai Yu , Liu Hsiao‐Sheng
Published: May 2009

Enterovirus 71 (EV71) is an important pathogen causing death in children under 5 years old worldwide. However, the underlying pathogenesis remains unclear. This study reveals that EV71 infection in rhabdomyosarcoma (RD) and neuroblastoma (SK‐N‐SH) cells stimulated the autophagic process, which was demonstrated by an increase of punctate GFP‐microtubule‐associated protein 1 light chain 3 (GFP‐LC3), the level of autophagosome‐bound LC3‐II protein and double‐membrane autophagosome formation. EV71‐induced autophagy benefited EV71 replication, which was confirmed by the autophagic inducer rapamycin and the inhibitor 3‐methyladenine. Signaling pathway investigation revealed that the decreased expression of phosphorylated mTOR and phosphorylated p70S6K is involved in EV71‐induced autophagy in a cell‐specific manner. The expression of phosphorylated extracellular signal‐regulated kinase (Erk) was suppressed consistently in EV71‐infected cells. However it did not participate in the autophagic response of the cell. Other signaling pathway molecules, such as Erk, PI3K/Akt, Bcl‐2, BNIP3, and Beclin‐1 were not affected by infection with EV71. Electron microscopy showed co‐localization of autophagosome‐like vesicles with either EV71‐VP1 or LC3 protein in neurons of the cervical spinal cord in ICR mice infected with EV71. In conclusion, EV71 infection triggered autophagic flux and induced autophagosome formation both in vitro and in vivo. Autophagy induced by EV71 is beneficial for viral replication. Understanding the role of autophagy induced by EV71 in vitro and the formation of autophagosome‐like vesicle in vivo provide new insights into the pathogenesis of EV71 infection. J. Med. Virol. 81:1241–1252, 2009. © 2009 Wiley‐Liss, Inc.
The dominance of China 1 in the spectrum of Epstein–Barr virus strains from Cantonese patients with nasopharyngeal carcinoma
Authors: Li Da‐Jiang , Bei Jin‐Xin , Mai Shi‐Juan , Xu Jin‐Fen , Chen Li‐Zhen , Zhang Ru‐Hua , Yu Xing‐Juan , Hong Ming‐Huang , Zeng Yi‐Xin , Kang Tiebang
Published: May 2009

Nasopharyngeal carcinoma is a disease with a remarkable geographic and ethnic distribution, and has a high incidence in southern China. Infection with Epstein–Barr virus (EBV) is an important contributing factor. The profile of EBV strains in Cantonese patients from Guangdong, the nasopharyngeal carcinoma endemic region in southern China, is described on the sequence variations in latent membrane protein 1 carboxyl‐terminus. The results show that China 1 was the dominant EBV strain detected in both the tumor biopsies and samples of throat washings, whereas multiple strains, including China 1, China 2, B95‐8, and Med, were detected in blood samples. In addition, a new strain named China 4 was found in blood samples. These findings suggest that the host population is susceptible to the predominant China 1 strain in the nasopharyngeal carcinoma endemic region of China, but its relationship with the host remains to be characterized further. J. Med. Virol. 81:1253–1260, 2009. © 2009 Wiley‐Liss, Inc.
Effect of HIV co‐infection on mutation patterns of HBV in patients with lamivudine‐resistant chronic hepatitis B
Authors: Iacomi Fabio , Vincenti Donatella , Vairo Francesco , Solmone Mariacarmela , Mariano Andrea , Piselli Pierluca , Puro Vincenzo , Capobianchi Maria Rosaria , Antonucci Giorgio
Published: May 2009

A retrospective review was performed comparing lamivudine‐resistance mutation patterns between patients infected with hepatitis B virus (HBV) with or without human immunodeficiency virus (HIV) co‐infection. Medical records that included a genotypic test of patients infected with HBV and treated with lamivudine as the only anti‐HBV drug were reviewed. Pol gene mutations were assessed by direct sequencing of the reverse transcriptase fragment 125–213 aa. Eighty‐nine patients infected with HBV (29 co‐infected with HIV) with rtM204V or rtM204I mutations were included. Multiple mutations associated with the YMDD motif were observed in 33 (55%) of 60 patients infected with HBV only and in 28 (96.6%) of patients co‐infected with HIV/HBV. In this latter group, the prevalence of the rtV173L + rtL180M + rtM204V triple mutation was 31% versus a prevalence of 3.4% observed among patients infected with HBV only. All patients with the triple mutational pattern showed sE164D + sI195M changes in the envelope gene. Multivariate analysis demonstrated that HIV co‐infection (adjusted OR 11.2, 95% CI 2.0–61.0) and HBV genotype A (adjusted OR 7.2, 95% CI 1.5–34.8) were the only independent variables associated with the chance of harboring rtM204V. Patients with HBV genotype A or HIV co‐infection were more likely to harbor the rtM204V mutation. Patients co‐infected with HIV showed multiple mutations more frequently, including the triple mutation that may elicit a vaccine escape phenotype. Among patients co‐infected with HIV/HBV, strict HBV DNA monitoring is essential to detect treatment failure and adapt therapy to avoid limitations of future therapeutic options or the emergence of a public health threat. J. Med. Virol. 81:1151–1156, 2009. © 2009 Wiley‐Liss, Inc.
Sequence and structural analysis of the 5′ noncoding region of hepatitis C virus in patients with chronic infection
Authors: Araújo Flávio Marcos Gomes , Machado‐Lima Ariane , Durham Alan Mitchell , Teixeira Rosangela , Oliveira Guilherme
Published: May 2009

Hepatitis C virus (HCV), exhibits considerable genetic diversity, but presents a relatively well conserved 5′ noncoding region (5′ NCR) among all genotypes. In this study, the structural features and translational efficiency of the HCV 5′ NCR sequences were analyzed using the programs RNAfold, RNAshapes and RNApdist and with a bicistronic dual luciferase expression system, respectively. RNA structure prediction software indicated that base substitutions will alter potentially the 5′ NCR structure. The heterogeneous sequence observed on 5′ NCR led to important changes in their translation efficiency in different cell culture lines. Interactions of the viral RNA with cellular transacting factors may vary according to the cell type and viral genome polymorphisms that may result in the translational efficiency observed. J. Med. Virol. 81:1212–1219, 2009. © 2009 Wiley‐Liss, Inc.
A neuropsychological study comparing patients infected with HCV and HBV without psychiatric comorbidities
Authors: Quarantini Lucas C. , Miranda‐Scippa Angela , Batista‐Neves Susana , Powell Vania B. , Abreu Neander , Abreu Katiusha C. , Moura Ilka , Crane Jacquelyn , Sampaio Aline S. , Netto Liana R. , deOliveira Irismar R. , Paraná Raymundo , Bressan Rodrigo A. , Lacerda Acioly L.T.
Published: May 2009

Hepatitis C is one of the most common chronic infectious diseases worldwide, with well‐documented extra‐hepatic manifestations, such as a broad number of cognitive deficits. These impairments may be explained by psychiatric comorbidities, which have not been investigated properly in the literature. In order to elucidate a specific hepatitis C virus (HCV) induced cognitive impairment not related to mental disorders, neuropsychological performance of patients infected with HCV was compared with that of patients infected with hepatitis B virus cognitive impairment, especially psychiatric comorbidities. A total of 33 patients infected with HCV and 22 patients infected with HBV were included in the study. There were no significant differences between the two groups with regard to age or years of education. The group of patients infected with HCV performed significantly worse on visuo‐spatial memory tasks after adjusting for years of education and age. There were no significant differences between patients infected with HCV and patients infected with HBV with regards to other neuropsychological functions. The data indicate that patients infected with HCV patients have poorer visuo‐spacial memory performance than patients infected with HBV, suggesting that the cognitive deficit may be specific to HCV infection and not to secondary comorbid psychiatric disorders. J. Med. Virol. 81:1184–1188, 2009. © 2009 Wiley‐Liss, Inc.
Tracking the transmission routes of genogroup II noroviruses in suspected food‐borne or environmental outbreaks of gastroenteritis through sequence analysis of the P2 domain
Authors: Xerry Jacqueline , Gallimore Chris I. , Iturriza‐Gómara Miren , Gray Jim J.
Published: May 2009

The aim of this study was to apply sequence analysis of a hyper variable region of the norovirus (NoV) genome in order to identify point source outbreaks associated with suspect food or water. The hyper‐variable region of the gene encoding the P2 domain was chosen as small differences in sequence are likely to indicate virus from different sources whereas identical sequence may reveal transmission routes and the source of contamination. Strains with 100% similarity were considered as originating from a common source, whereas, strains with one or more mutations in the hyper variable region sequenced were regarded as representing unrelated transmission events. This study was able to identify a point source outbreak of a dominant strain, GII‐4, on a cruise ship but also of a less common strain, GII‐2, between two schools. Also identical GII‐3 strains were demonstrated in food handlers amongst the same outbreak; however epidemiologically related outbreaks showed different GII‐3 strains indicating multiple sources of contamination. J. Med. Virol. 81:1298–1304, 2009. © 2009 Wiley‐Liss, Inc.