bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic value of microsatellite instability determined by immunohistochemical staining of hMSH2 and hMSH6 in urothelial carcinoma of the bladder
Authors: MYLONA E. , ZAROGIANNOS A. , NOMIKOS A. , GIANNOPOULOU I. , NIKOLAOU I. , ZERVAS A. , NAKOPOULOU L.
Published: January 2008

Mismatch repair (MMR) genes are involved in the recognition and repair of acquired DNA damage, which arises during cell division, thus playing an essential role in preserving genetic stability. Immunohistochemistry was applied to 130 specimens from urothelial carcinoma (UC) of the bladder to detect expression of MMR gene products hMSH2 and hMSH6, and to investigate its clinicopathological and prognostic value. hMSH2 and hMSH6 protein expression was exclusively detected in the nuclei of malignant cells. Of the 112 cases evaluable for hMSH2, 29 (25.9%) were negative and of the 130 UCs evaluable for hMSH6, 64 (49.2%) were negative, and were thus considered to depict MSI. Nuclear hMSH2 values were statistically lower in non‐invasive UCs (Ta‐T1) (p=0.013) and in carcinomas with decreased p53 staining (p=0.04). Lower hMSH6 values were more often met in well‐differentiated tumors (p<0.0001) and in tumors with low expression of p53 (p=0.016), topoIIα and caspase 3 (p=0.017 and p=0.018, respectively). Both hMSH2‐ and hMSH6‐negative immunoreactions were found to have a favorable impact on overall patient survival (p=0.041 and p=0.034, respectively), this finding being further verified in the multivariate analysis of hMSH2 (p=0.026). This is the first study to show that lack (and not reduction designated according to various cut‐off points) of hMSH2 and hMSH6 correlated with non‐invasive tumors of lower grade and is of favorable prognostic significance in patients suffering from bladder carcinoma.
Prostate‐specific antigen‐positive extramammary Paget's disease––association with prostate cancer
Authors: HAMMER ANNE , HAGER HENRIK , STEINICHE TORBEN
Published: January 2008

Extramammary Paget's disease (EMPD) is a rare intraepidermal adenocarcinoma that primarily affects the anogenital region. Cases of EMPD reacting with PSA (prostate‐specific antigen) have previously been associated with underlying prostate cancer. However, a recent case of EMPD in our department has led us to question the value of PSA as an indicator of underlying prostate cancer. Clinical and pathological data were obtained for 16 cases of EMPD. Formalin‐fixed, paraffin‐embedded tissue blocks from the primary skin lesions were investigated using PSA and other immunohistochemical markers. 5 of the 16 cases of EMPD stained positive for PSA (2 women and 3 men). However, no reactivity was seen for the prostatic marker P501S. Three of the five patients had been diagnosed with internal malignant disease–two with prostate cancer, stage 1. Immunohistochemical investigations of the tumour specimens from the prostate revealed an immunoprofile which was very different from that of the primary skin lesion. In our study, no cases of EMPD with PSA positivity seem to represent an extension of an underlying prostatic adenocarcinoma. PSA positivity can be seen in cases of EMPD without associated adenocarcinoma of the prostate.
EGFR expression in urothelial carcinoma of the upper urinary tract is associated with disease progression and metaplastic morphology
Authors: LEIBL SEBASTIAN , ZIGEUNER RICHARD , HUTTERER GEORG , CHROMECKI THOMAS , REHAK PETER , LANGNER CORD
Published: January 2008

EGFR represents a promising therapeutic target in urothelial cancer (UC). Our study aimed to investigate the clinicopathological significance of EGFR in upper urinary tract UC. EGFR was immunohistochemically assessed (EGFR pharmDX kitTM) in 268 consecutive tumours using a tissue microarray technique and correlated with other histopathological parameters as well as patient outcome. EGFR immunoreactivity was observed in 140/253 (55%) evaluable UCs and was associated with high tumour stage (47% pTa/pT1 vs 66% pT2‐pT4; p=0.003) and high tumour grade (45% low grade vs 67% high grade; p<0.001). In addition, EGFR expression was associated with metaplastic squamous and/or glandular differentiation (p<0.001). EGFR staining intensity was 1+ in 49%, 2+ in 31%, and 3+ in 20% of cases. EGFR 3+ staining intensity was associated with the occurrence of metastatic disease by univariate analysis (p=0.016). Multivariate analysis, however, proved only pT stage >1 (p<0.001) and high tumour grade (p<0.001) to be independent predictors of patient outcome. In conclusion, EGFR was significantly associated with advanced disease and metaplastic squamous and/or glandular differentiation. Since UCs with metaplastic morphology have been shown to be more resistant to conventional radiotherapy or chemotherapy, the strikingly strong EGFR expression in these tumours may offer a new perspective for affected patients.
‘HPV vulvitis’ revisited: frequent and persistent detection of novel epidermodysplasia verruciformis‐associated HPV genotypes
Authors: Lin Ming‐Tseh , Rohwedder Angela , Mysliborski Judith , Leopold Kathryn , Wilson Vincent L. , Carlson J. Andrew
Published: January 2008

Background:  ‘Human papillomavirus (HPV) vulvitis’ is a disputed entity where most studies examining for genital‐mucosal (GM) HPV have been negative.

Methods:  Using degenerate and type specific primers for cutaneous (CU), GM and epidermodysplasia verruciformis (EV) HPV types, the prevalence of specific HPV types was investigated in biopsy specimens from 19 women with ‘HPV vulvitis’, seven with asymptomatic vulvar squamous papillomatosis (ASxVSP), and controls of vulvar fibroepithelial polyps (FEP) (15), vulvar condyloma (10) and normal vulva (NV) (10).

Results:  HPV DNA/EV HPV/GM HPV/CU HPV were detected in 84/74/47/5% of vulvitis patients, 78/71/0/28% of ASxVSP, 47/20/20/7% of FEP, 10/10%/0/0 of NV and 100/0/100/10% of condyloma. Fourteen putatively novel HPV genotypes were detected in vulvitis and ASxVSP patients, but not in controls. The two most frequent novel EV HPV, Alb‐4 and DL285, were detected in 9/19 (47%) and 5/19 (26%) of vulvitis cases and were persistently identified in serial biopsies. HPV co‐infection and Alb‐4 infection occurred significantly more frequently in vulvitis patients, particularly those complaining of ‘burning’ (62/62% vs. 17/7%, p ≤ 0.004). Koilocytosis was identified significantly more frequently in vulvitis compared with non‐condyloma controls (81% vs. 40%, p = 0.0001), and its presence correlated with detection of HPV DNA (r = 0.3, p = 0.002).

Conclusion:  The high frequency of novel EV HPV in HPV vulvitis and correlation of clinicopathologic findings with HPV DNA suggests that HPV vulvitis may indeed exist.
The 3G5 antigen is expressed in dermal mast cells but not pericytes
Authors: Gushi Akira , Tanaka Masashi , Tsuyama Shinichiro , Nagai Taku , Kanzaki Tamotsu , Kanekura Takuro , Matsuyama Takami
Published: January 2008

Background:  It has been shown that the 3G5 antigen recognized by monoclonal antibody 3G5 (mAb 3G5) is a useful marker of pericytes in normal human skin. However, most 3G5 antigen‐expressing cells in capillary vessels were stained negatively for α‐smooth muscle actin (α‐SMA), a prominent pericyte marker. This study was designed to determine whether the expression of the 3G5 antigen is restricted to specific stages of pericyte development, or if it is expressed in other cells rather than pericytes in capillary vessels.

Methods:  3G5 antigen‐expressing cells were detected in normal human skin, granulating tissues from decubitus ulcers and inflammatory psoriatic skin with extensive angiogenesis using double immunofluorescent staining with mAb 3G5 and monoclonal antibodies (mAbs) to various pericyte markers, tryptase and chymase. Furthermore, using immunoelectron microscopy, 3G5 antigen‐expressing cells were observed in the granulating tissues.

Results:  The immunoelectron microscopic findings and double immunofluorescent staining (using mAb 3G5 and either anti‐tryptase or anti‐chymase mAbs) showed that 3G5 antigen‐expressing cells were mast cells in normal skin, granulating tissues and psoriatic skin.

Conclusions:  The results indicated that 3G5 antigen is a marker of mast cells, but not of pericytes in normal and diseased skin.
Association between promoter and coding region mutations of UDP‐glucuronosyltransferase 1A1 and β‐thalassemia/Hb E with cholelithiasis
Authors: Tankanitlert Jeeranut , Morales Noppawan P. , Fucharoen Pranee , Fucharoen Suthat , Chantharaksri Udom
Published: January 2008

Background and objectives:  Cholelithiasis has been observed with high incidence in beta‐thalassemia/hemoglobin E (β‐thal/Hb E). Recent studies have shown that a variant TATA‐box in the promoter region of the UDP‐glucuronosyltransferase 1A1 (UGT1A1) gene is associated with the development of cholelithiasis. The coding region mutation (G71R) of the UGT1A1 gene was higher in Asians than those in Caucasians. The relationship between the variant UGT1A1 promoter and coding region gene and cholelithiasis in β‐thal/Hb E subjects were investigated.

Methods:  One hundred and seventeen β‐thal/Hb E subjects entered this study. The TATA‐box and G71R mutations were analyzed by fragment size analysis and restriction fragment length polymorphism methods, respectively.

Results:  The incidence of cholelithiasis was higher in heterozygous (68.3%) and homozygous (100%) subjects compared with normal UGT1A1 haplotype (61.4%). Total bilirubin level (6.0 ± 2.03 mg/dL) in the homozygous group was significantly higher than that of wild type (3.31 ± 1.83 ng/dL). Prevalence of cholelithiasis increased with age (OR = 1.1, 95% CI = 1.03–1.12, P < 0.001). Female gender (OR = 3.7, 95% CI = 1.3–10.6, P < 0.01) and elevated liver enzyme (OR = 1.02, 95%CI = 1.0–1.04, P < 0.02) were two other risk factors for cholethiasis in β‐thal/Hb E.

Conclusion:  This study shows that the combined TATA‐box variants and G71R mutations of the UGT1A1 is associated with cholelithiasis in β‐thal/Hb E.
Characterization of a Listeria monocytogenes Protein Interfering with Rab5a
Authors: Alvarez‐Dominguez Carmen , Madrazo‐Toca Fidel , Fernandez‐Prieto Lorena , Vandekerckhove Joël , Pareja Eduardo , Tobes Raquel , Gomez‐Lopez Maria Teresa , Cerro‐Vadillo Elida Del , Fresno Manuel , Leyva‐Cobián Francisco , Carrasco‐Marín Eugenio
Published: January 2008

Listeria monocytogenes (LM) phagocytic strategy implies recruitment and inhibition of Rab5a. Here, we identify a Listeria protein that binds to Rab5a and is responsible for Rab5a recruitment to phagosomes and impairment of the GDP/GTP exchange activity. This protein was identified as a glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) from Listeria (p40 protein, Lmo 2459). The p40 protein was found within the phagosomal membrane. Analysis of the sequence of LM p40 protein revealed two enzymatic domains: the nicotinamide adenine dinucleotide (NAD)‐binding domain at the N‐terminal and the C‐terminal glycolytic domain. The putative ADP‐ribosylating ability of this Listeria protein located in the N‐terminal domain was examined and showed some similarities to the activity and Rab5a inhibition exerted by Pseudomonas aeruginosa ExoS onto endosome–endosome fusion. Listeria p40 caused Rab5a‐specific ADP ribosylation and blocked Rab5a‐exchange factor (Vps9) and GDI interaction and function, explaining the inhibition observed in Rab5a‐mediated phagosome–endosome fusion. Meanwhile, ExoS impaired Rab5‐early endosomal antigen 1 (EEA1) interaction and showed a wider Rab specificity. Listeria GAPDH might be the first intracellular gram‐positive enzyme targeted to Rab proteins with ADP‐ribosylating ability and a putative novel virulence factor.
Dynamin‐Dependent Biogenesis, Cell Cycle Regulation and Mitochondrial Association of Peroxisomes in Fission Yeast
Authors: Jourdain Isabelle , Sontam Dharani , Johnson Chad , Dillies Clément , Hyams Jeremy S.
Published: January 2008

Peroxisomes were visualized for the first time in living fission yeast cells. In small, newly divided cells, the number of peroxisomes was low but increased in parallel with the increase in cell length/volume that accompanies cell cycle progression. In cells grown in oleic acid, both the size and the number of peroxisomes increased. The peroxisomal inventory of cells lacking the dynamin‐related proteins Dnm1 or Vps1 was similar to that in wild type. By contrast, cells of the double mutant dnm1Δ vps1Δ contained either no peroxisomes at all or a small number of morphologically aberrant organelles. Peroxisomes exhibited either local Brownian movement or longer‐range linear displacements, which continued in the absence of either microtubules or actin filaments. On the contrary, directed peroxisome motility appeared to occur in association with mitochondria and may be an indirect function of intrinsic mitochondrial dynamics. We conclude that peroxisomes are present in fission yeast and that Dnm1 and Vps1 act redundantly in peroxisome biogenesis, which is under cell cycle control. Peroxisome movement is independent of the cytoskeleton but is coupled to mitochondrial dynamics.
How Many Ways Through the Golgi Maze?
Authors: Prydz Kristian , Dick Gunnar , Tveit Heidi
Published: January 2008

The secretory route in eukaryotic cells has been regarded as one common pathway from the endoplasmic reticulum (ER) through the Golgi cisternae to the trans Golgi network where recognition, sorting and exit of cargo molecules are thought to occur. Morphologically, the ribosome‐coated ER is observed throughout the cytoplasm, while the Golgi apparatus usually is confined to a perinuclear position in mammalian cells. However, Golgi outposts have been observed in neuronal dendrites and dispersed Golgi elements in skeletal muscle myofibers. In insects, like in Drosophila melanogaster imaginal disc cells and epidermal cells of Tobacco and Arabidopsis leafs, individual Golgi stacks are distributed throughout the cytoplasm. Golgi stacks do not only differ in their intracellular localization but also in the number of stacks from one to several hundreds. Each stack consists of closely aligned, flattened, membrane‐limited cisternae. The number of cisternae in a Golgi stack is also variable, 2–3 in some ciliates, 10 in many plant cell types and up to 30 in certain euglenoids. The yeast Saccharomyces cerevisiae has a Golgi structure of minimal complexity with scattered solitary cisternae. It is assumed that the number of Golgi cisternae reflects the overall complexity of the enzymatic reactions that occur in their lumen, while the number of stacks reflects the load of macromolecules arriving at the cis side. In this review, we will focus on how the available morphological and biochemical data fit with the current view of protein sorting in the secretory pathway, particularly in polarized cells like neuronal and epithelial cells.
ITSN‐1 Controls Vesicle Recycling at the Neuromuscular Junction and Functions in Parallel with DAB‐1
Authors: Wang Wei , Bouhours Magali , Gracheva Elena O. , Liao Edward H. , Xu Keli , Sengar Ameet S. , Xin Xiaofeng , Roder John , Boone Charles , Richmond Janet E. , Zhen Mei , Egan Sean E.
Published: January 2008

Intersectins (Itsn) are conserved EH and SH3 domain containing adaptor proteins. In Drosophila melanogaster, ITSN is required to regulate synaptic morphology, to facilitate efficient synaptic vesicle recycling and for viability. Here, we report our genetic analysis of Caenorhabditis elegans intersectin. In contrast to Drosophila, C. elegans itsn‐1 protein null mutants are viable and display grossly normal locomotion and development. However, motor neurons in these mutants show a dramatic increase in large irregular vesicles and accumulate membrane‐associated vesicles at putative endocytic hotspots, approximately 300 nm from the presynaptic density. This defect occurs precisely where endogenous ITSN‐1 protein localizes in wild‐type animals and is associated with a significant reduction in synaptic vesicle number and reduced frequency of endogenous synaptic events at neuromuscular junctions (NMJs). ITSN‐1 forms a stable complex with EHS‐1 (Eps15) and is expressed at reduced levels in ehs‐1 mutants. Thus, ITSN‐1 together with EHS‐1, coordinate vesicle recycling at C. elegans NMJs. We also found that both itsn‐1 and ehs‐1 mutants show poor viability and growth in a Disabled (dab‐1) null mutant background. These results show for the first time that intersectin and Eps15 proteins function in the same genetic pathway, and appear to function synergistically with the clathrin‐coat‐associated sorting protein, Disabled, for viability.
Adverse health effects related to tobacco smoke exposure in a cohort of three‐year olds
Authors: Johansson AnnaKarin , Ludvigsson Johnny , Hermansson Göran
Published: January 2008

Aim: To analyse the importance of mothers' smoking during pregnancy and/or environmental tobacco smoke (ETS) exposure in early childhood for children's health and well‐being at the age of 3 years.

Methods: Four groups from a population based cohort (n = 8850) were compared: children with nonsmoking mother during pregnancy and nonsmoking parents at the age of 3 years (n = 7091); children with only foetal exposure (n = 149); children exposed only postnatally (n = 895) and children exposed both pre‐ and postnatally (n = 595). Odds ratios and 95% confidence intervals were calculated.

Results: Children exposed both pre‐ and postnatally had more wheezing (1.14; 1.07–1.21) and rhinitis (1.16; 1.06–1.26), used more cough‐mixture (1.07; 1.01–1.14) and broncodilatating drugs (1.08; 1.02–1.15) and suffered more from excessive crying (1.31; 1.13–1.51) and irritability (1.27; 1.09–1.48) compared to children with nonsmoking parents. Children exposed only postnatally had more rhinitis (1.24; 1.12–1.37), used more cough‐mixture (1.14; 1.05–1.29) and suffered more from poor sleep (1.26; 1.07–1.47) than children of nonsmoking parents. Children with prenatal exposure only used more broncodilatating drugs (1.45; 1.03–2.04) and suffered more from poor sleep (2.06; 1.09–3.87).

Conclusion: Health differences, small but significant, indicate that prenatal and/or postnatal ETS exposure alone, or in combination, seems to interfere with child health, supporting the importance of zero tolerance. However, as most smoking parents in Sweden try to protect their children from ETS exposure, the results also might indicate that protective measures are worthwhile.
A genetic cause for neonatal encephalopathy: incontinentia pigmenti with NEMO mutation
Authors: Loh Ne‐Ron , Jadresic Lyda P , Whitelaw Andrew
Published: January 2008

Incontinentia pigmenti (IP) is not generally recognized as a cause of neonatal encephalopathy. A full‐term infant developed a rash and encephalopathy with lesions in the basal ganglia and periventricular white matter 3 days after a normal delivery. Typical skin changes of IP were confirmed by histology and mutation analysis of the NFκB essential modulator (NEMO) gene.

Conclusion: The mechanism of brain injury appears to be increased apoptosis after inflammation and this condition should be included in differential diagnosis of neonatal encephalopathy if skin lesions are present.
Maternal chronic hepatitis B virus does not affect neonatal biotinidase activity
Authors: Vlachos George D , Schulpis Kleopatra H , Papakonstantinou Evangelos , Papadakis Miltiadis , Elefsiniotis Ioannis , Papassotiriou Ioannis , Antsaklis Aris
Published: January 2008

Backround: Biotinidase activity is closely related to liver function.

Aim: To evaluate whether maternal chronic hepatitis B virus (HBV) infection affects neonatal biotinidase activity.

Patients and Methods: Twenty‐three asymptomatic pregnant women with HBV (group A) and 28 healthy pregnant women (controls) in the delivery room and their newborns (cord blood) underwent laboratory examinations. Serological HBV and liver function tests were performed with standard techniques, while biotinidase activity was measured with an HPLC method.

Results: Serological HBV tests and HBV DNA showed chronic HBV (precore mutant G1896A) in group A, whereas anti‐HBc and anti‐HBe were detected in their neonates. Liver function chemistry was found normal in controls and both groups of newborns. Moderately increased transaminases were found in the infected mothers. Interestingly, albumin levels did not differ among the studied groups. Biotinidase activity in HBV mothers (5.76 ± 0.6 nmol/min/mL) was significantly decreased (p < 0.001) as compared to controls (8.43 ± 0.65 nmol/min/mL). The enzyme activity did not differ among the neonates. Biotinidase activity inversely correlated with transaminases but not with albumin or with HBV‐DNA levels.

Conclusions: Decreased biotinidase activities were evaluated in mothers with HBV and normal in their neonates. Biotin supplementation in the diseased mothers may prevent possible symptoms due to biotin recycling impairment.
Influence of Interimplant Distance on Bone Microstructure: A Histomorphometric Study in Dogs
Authors: Traini Tonino , Novaes Arthur B. , Papalexiou Vula , Piattelli Adriano
Published: January 2008

The microstructure of the crestal alveolar bone is important for both the maintenance of osseointegration and the location of the gingival soft tissues. The aim of this study was to evaluate and compare the bone microstructure of the alveolar bone and of the interimplant bone in implants inserted at different interimplant distances. The mandibular bilateral premolars of six dogs were extracted, and after 12 weeks, each dog received eight implants, for a total of 48 implants. Two pairs of implants, one for each hemiarch, were separated by 2 mm (group 1) and by 3 mm (group 2). After 12 weeks, the implants received temporary acrylic prostheses. After four more weeks, metallic crowns substituted the temporary prostheses. After an additional 8 weeks the animals were sacrificed and the hemimandibles were removed, dissected, and processed. The longitudinal collagen fiber orientation was 43.2% for the alveolar bone; it was 30.3% for the 2‐mm group and 43.9% for the 3‐mm group. There was a statistically significant difference between the 2‐mm and 3‐mm groups (p < .05). The orientation of transverse collagen fibers was 47.8% for the alveolar bone; it was 37.3% for the 2‐mm group and 56.3% for the 3‐mm group. There was a statistically significant difference between the 2‐mm and 3‐mm groups (p < .05). The marrow spaces were 34.87% for the alveolar bone, 52.3% for the 2‐mm group, and 59.9% for the 3‐mm group. There was a statistically significant difference between the alveolar bone and the 3‐mm group (p < .05). The low mineral density index was 36.29 for the alveolar bone, 46.76 for the 2‐mm group, and 17.91 for the 3‐mm group. There was a statistically significant difference between the 2‐mm and 3‐mm groups (p < .05). The high mineral density was 87.57 for the alveolar bone, 72.58 for the 2‐mm group, and 84.91 for the 3‐mm group. There was a statistically significant difference between the alveolar bone and the 2‐mm group (p < .05). The collagen fiber orientation resulted in statistically significant differences in both the 2‐mm and 3‐mm groups compared with the alveolar bone. The marrow spaces appeared significantly increased in the 3‐mm group compared with the alveolar bone. The low mineral density index was significantly higher in the 2‐mm group, while the high mineral density index was significantly higher in the alveolar bone. In conclusion, the interimplant distance should not be less than 3 mm.
Evaluations of Bone Tissue Integration to Pure and Alloyed Titanium Implants
Authors: Stenport Victoria Franke , Johansson Carina B.
Published: January 2008

Purpose: This study was performed for comparisons of tissue integration to commercially pure (CP) and titanium‐6‐aluminum‐4 vanadium (Ti‐6‐Al‐4V) implants using various existing three‐dimensional biomechanical and two‐dimensional histomorphometrical techniques, and to monitor the loosening torque during in vivo removal torque (RTQ) test with a novel unit not used before in a pilot study in rabbits.

Materials and Methods: The implants were topographically characterized and inserted in femurs and tibiae of five rabbits (in total 40 implants, 20 per group). After 16 weeks, the implant integration was biomechanically evaluated by: (1) resonance frequency test, and (2) peak RTQ test and the graph from the monitoring curve. Biopsies of the implants in situ were processed to undecalcified cut and ground sections followed by light microscopical quantifications. Shear strength calculations were performed.

Results: Significantly higher mean value of RTQ (p = .01) and shear strength tests (p = .03) were observed for the CP titanium implants compared to Ti‐6‐Al‐4V implants. The monitoring curve from the RTQ test demonstrated no differences in the shape or form that could provide further information about the differences in the implant‐to‐bone attachment.

Conclusions: The CP titanium implants showed increased RTQ and shear strength values compared to the Ti‐6‐Al‐4V implants. The new tool of monitoring the RTQ curve could not demonstrate differences between the two materials. The exact influence of the implant materials on the surrounding tissues needs to be further investigated.
The Bone Tissue Responses to Prehydrated and Collagenated Cortico‐Cancellous Porcine Bone Grafts: A Study in Rabbit Maxillary Defects
Authors: Nannmark Ulf , Sennerby Lars
Published: January 2008

Background: Bone substitutes should have osteoconductive properties and be completely replaced with new bone with time. Adding collagen gel to prehydrated and collagenated porcine bone (PCPB) particles results in a sticky and moldable material which facilitates clinical handling. However, the possible influence of the gel on the bone tissue response is not known.

Purpose: The objective of the study was to evaluate the bone tissue responses to PCPB graft with or without collagen gel and to evaluate the resorption/degradation properties of the biomaterials.

Materials and Methods: Fourteen rabbits were used in the study. Bilateral bone defects, 5 × 8 × 3 mm, were created in the maxilla and filled with PCPB + collagen gel (test) or with PCPB only (control) and covered with a collagen membrane. Animals were killed after 2 (n = 3), 4 (n = 3), and 8 weeks (n = 8) for histological and morphometrical evaluations.

Results: There were no differences between test and control defects. Both materials showed bone formation directly on the particles by typical osteoblastic seams. The bone area increased with time (2–8 weeks) for both sides, from 16.2% (control) and 19.2% (test) to 42.7 and 43.8%, respectively. The PCPB, whether mixed with collagen gel or not, was resorbed by osteoclasts as well as part of remodeling with the formation of osteons within the particles. Morphometry showed a decrease of PCPB area from 19.4% (control) and 23.8% (test) after 2 weeks to 3.7 and 9.3% after 8 weeks, respectively.

Conclusions: Mixing collagen gel and PCPB to facilitate the clinical handling does not influence the bone tissue responses to the material, which exhibited osteoconductive properties and was resorbed with time.
Complications of Computer‐Aided‐Design/Computer‐Aided‐Machining‐Guided (NobelGuide™) Surgical Implant Placement: An Evaluation of Early Clinical Results
Authors: Yong Loong Tee , Moy Peter K.
Published: January 2008

Purpose: The aim of this study was to evaluate early clinical results of computer‐aided design (CAD)/computer‐aided machining (CAM)‐guided surgical implant placement (NobelGuide™, Nobel Biocare, Yorba Linda, CA, USA) with focus on surgical and/or prosthetic complications, management, and prevention.

Materials and Methods: Thirteen patients rehabilitated between March 2003 and October 2006 with CAD/CAM‐guided dental implants and immediate loading (NobelGuide, Nobel Biocare) were evaluated. The treatment planning and procedures were carried out in accordance to the system protocol. The complications encountered in this case series were classified and assessed according to early (planning and procedural – surgical; prosthetic) and late complications (surgical; prosthetic).

Results: The prosthetic complications outnumbered surgical complications both in the early and late treatment phases. The main early surgical complication was bony interference that prevented complete seating of the prostheses. Most of the late surgical complications were implant failures with an overall failure rate of 9%. Fracture of the carbon fiber framework prosthesis was the main late prosthetic complication.

Conclusions: The NobelGuide system is a reliable treatment modality, but not without its complications. Strict adherence to the system protocol is the key prevention of complications.
The Virginal Conception and Its Meanings
Authors: O'Collins Gerald
Published: January 2008

In Jeffrey Archer's The Gospel According to Judas, Judas dismisses the virginal conception of Jesus as no more than another example of ‘Greek myths that tell of gods in heaven who produce offspring following a union with women of this earth’. To attribute such a view to a first-century Jew like Judas seems strange, since the earliest evidence shows Jewish critics of the Christian movement rejecting the virginal conception as a case of illegitimacy. In any case such Greek myths do not provide plausible sources for the two Gospel accounts of the virginal conception. Yet such merely historical debate is insufficient. One should press on to illustrate the religious significance and theological importance of the virginal conception within the whole story of Jesus: for instance, the role of this conception in revealing the Trinity at work for human salvation.
Restoration and Eighteenth‐Century Drama: New Directions in the Field
Authors: Rosenthal Laura J.
Published: January 2008

This essay offers an overview of recent critical books published on Restoration and eighteenth‐century drama, with a glance back to the critical history of this field. The essay emphasizes critical explorations of the earlier part of the period and provides a bibliography for further reading.
Contesting Literary Biography in the Romantic Period: The Foreshadowing of Psychological Biography
Authors: Darcy Jane
Published: January 2008

This essay surveys critical approaches to Romantic literary biography, both by Romantic writers themselves and by recent scholars. While accepting the argument that an epistemological shift in the late eighteenth century saw the emergence of interest in subjective experience as recorded in the new genre of full‐length biography, it rejects the centrality some scholars have placed on the debate over whether literary biography was an ‘art’ rather than a ‘science’. Rather it suggests that the central debate was over the depiction of the ‘inner man’. Does the imperative for biographical ‘truth’ sanction the publication of revelations – often scandalous or shocking – of a subject's private life? Or is the ‘truth’ to be found in Carlyle's insistence on the transcendent powers of the visionary poet? Key biographies will be considered, including those of Gray, Burns, Cowper and Byron, as will key biographers, including Mason, Hayley, Currie, Lockhart, Southey, Moore and Carlyle. The relative importance of biographical commentators James Stanfield and Isaac D’Israeli forms an important part of this discussion. D’Israeli's introduction from his reading of the new German psychology of the term ‘psychological character’ is an important step on the royal road that will lead to Freudian biography.
Contributions to catalysis and potential interactions of the three catalytic domains in a contiguous trimeric creatine kinase
Authors: Hoffman Gregg G. , Davulcu Omar , Sona Sona , Ellington W. Ross
Published: January 2008

Three separate creatine kinase (CK) isoform families exist in animals. Two of these (cytoplasmic and mitochondrial) are obligate oligomers. A third, flagellar, is monomeric but contains the residues for three complete CK domains. It is not known whether the active sites in each of the contiguous flagellar domains are catalytically competent, and, if so, whether they are capable of acting independently. Here we have utilized site‐directed mutagenesis to selectively disable individual active sites and all possible combinations thereof. Kinetic studies showed that these mutations had minimal impact on substrate binding and synergism. Interestingly, the active sites were not catalytically equivalent, and were in fact interdependent, a phenomenon that has previously been reported only in the oligomeric CK isoforms.
Brox, a novel farnesylated Bro1 domain‐containing protein that associates with charged multivesicular body protein 4 (CHMP4)
Authors: Ichioka Fumitaka , Kobayashi Ryota , Katoh Keiichi , Shibata Hideki , Maki Masatoshi
Published: January 2008

Human Brox is a newly identified 46 kDa protein that has a Bro1 domain‐like sequence and a C‐terminal thioester‐linkage site of isoprenoid lipid (CAAX motif) (C standing for cysteine, A for generally aliphatic amino acid, and X for any amino acid). Mammalian Alix and its yeast ortholog, Bro1, are known to associate with charged multivesicular body protein 4 (CHMP4), a component of endosomal sorting complex required for transport III, via their Bro1 domains and to play roles in sorting of ubiquitinated cargoes. We investigated whether Brox has an authentic Bro1 domain on the basis of its capacity for interacting with CHMP4s. Both Strep Tactin binding sequence (Strep)‐tagged wild‐type Brox (Strep–BroxWT) and Strep‐tagged farnesylation‐defective mutant (Cys→Ser mutation; Strep–BroxC408S) pulled down FLAG‐tagged CHMP4b that was coexpressed in HEK293 cells. Treatment of cells with a farnesyltransferase inhibitor, FTI‐277, caused an electrophoretic mobility shift of Strep–BroxWT, and the mobility coincided with that of Strep–BroxC408S. The inhibitor also caused a mobility shift of endogenous Brox detected by western blotting using polyclonal antibodies to Brox, suggesting farnesylation of Brox in vivo. Fluorescence microscopic analyses revealed that Strep–BroxWT exhibited accumulation in the perinuclear area and caused a punctate pattern of FLAG–CHMP4b that was constitutively expressed in HEK293 cells. On the other hand, Strep–BroxC408S showed a diffuse pattern throughout the cell, including the nucleus, and did not cause accumulation of FLAG–CHMP4b. Fluorescent signals of monomeric green fluorescent protein (mGFP)‐fused BroxWT merged partly with those of Golgi markers and with those of abnormal endosomes induced by overexpression of a dominant negative mutant of AAA type ATPase SKD1/Vps4B in HeLa cells, but such colocalization was less efficient for mGFP–BroxC408S. These results suggest a physiological significance of farnesylation of Brox in its subcellular distribution and efficient interaction with CHMP4s in vivo.
Amyloid–cholinesterase interactions
Authors: Inestrosa Nibaldo C. , Dinamarca Margarita C. , Alvarez Alejandra
Published: January 2008

Acetylcholinesterase is an enzyme associated with senile plaques. Biochemical studies have indicated that acetylcholinesterase induces amyloid fibril formation by interaction throughout the peripherical anionic site of the enzyme forming highly toxic acetylcholinesterase–amyloid‐β peptide (Aβ) complexes. The pro‐aggregating acetylcholinesterase effect is associated with the intrinsic amyloidogenic properties of the corresponding Aβ peptide. The neurotoxicity induced by acetylcholinesterase–Aβ complexes is higher than the that induced by the Aβ peptide alone, both in vitro and in vivo. The fact that acetylcholinesterase accelerates amyloid formation and the effect is sensitive to peripherical anionic site blockers of the enzyme, suggests that specific and new acetylcholinesterase inhibitors may well provide an attractive possibility for treating Alzheimer’s disease. Recent studies also indicate that acetylcholinesterase induces the aggregation of prion protein with a similar dependence on the peripherical anionic site.
Biochemical characterization of fragmented human MCM2
Authors: Komamura‐Kohno Yuki , Tanaka Rikou , Omori Akira , Kohno Toshiyuki , Ishimi Yukio
Published: January 2008

The molecular dissection of human MCM2, a constituent of MCM2‐7 licensing factor complex, was performed to identify the region responsible for its biochemical activities. Partial digestion with trypsin dissected the MCM2 protein into a central region (148–676) containing ATPase motifs and a C‐terminal region (677–895). These two fragments, along with three other fragments (148–441, 442–676 and 442–895), were produced using the wheat germ cell‐free system and were examined for their ability to inhibit MCM4/6/7 helicase activity. Two fragments (442–895 and 677–895) containing the C‐terminus were partly inhibitory to the activity. Further dissection revealed that one fragment (713–895) has strong inhibitory activity. The inhibitory activity of the smaller fragments derived from the C‐terminal region correlated with their ability to inhibit SV40 T antigen helicase activity and also with their ability to bind to ssDNA, which has been shown by gel mobility shift analysis. These results strongly suggest that the MCM2 fragments derived from the C‐terminal region inhibit DNA helicase activity through their ability to bind to ssDNA. In contrast, two fragments (148–441 and 442–676) from the central region were mainly responsible for the interaction between MCM2 and MCM4, and this was revealed by a pulldown analysis using MCM4 protein beads. Finally, only complete MCM2, not the smaller fragments, could disassemble the MCM4/6/7 hexamer into the MCM2/4/6/7 tetramer.
Spo0B of Bacillus anthracis – a protein with pleiotropic functions
Authors: Mattoo Abid R. , Saif Zaman Mohd , Dubey Gyanendra P. , Arora Amit , Narayan Azeet , Jailkhani Noor , Rathore Kusum , Maiti Souvik , Singh Yogendra
Published: January 2008

Spo0B is an important component of the phosphorelay signal transduction pathway, the pathway involved in the initiation of sporulation in Bacillus subtilis. Bioinformatic, phylogenetic and biochemical studies showed that Spo0B of Bacillus anthracis has evolved from citrate/malate kinases. During the course of evolution, Spo0B has retained the characteristic histidine kinase boxes H, N, F, G1 and G2, and has acquired nucleotide‐binding domains, Walker A and Walker B, of ATPases. Owing to the presence of these domains, autophosphorylation and ATPase activity was observed in Spo0B of B. anthracis. Mutational studies showed that among the six histidine residues, His13 of the H‐box is involved in the autophosphorylation activity of Spo0B, whereas Lys33 of the Walker A domain is associated with the ATPase activity of the protein. Thermodynamic and binding studies of the binding of Mg‐ATP to Spo0B using isothermal titration calorimetry (ITC) suggested that the binding is driven by favorable entropy changes and that the reaction is exothermic, with an apparent dissociation constant (Kd) equal to 0.02 mm. The value of the dissociation constant (Kd = 0.05 mm) determined by the intrinsic fluorescence of trytophan of Spo0B was similar to that obtained by ITC studies. The purified Spo0B of B. anthracis also showed nucleoside diphosphate kinase‐like activity of phosphate transfer from nucleoside triphosphate to nucleoside diphosphate. This is the first evidence for Spo0B of B. anthracis as an enzyme with histidine kinase and ATPase activities, which may have important roles to play in sporulation and pathogenesis.
Abundance of intrinsic disorder in SV‐IV, a multifunctional androgen‐dependent protein secreted from rat seminal vesicle
Authors: Vilasi Silvia , Ragone Raffaele
Published: January 2008

The potent immunomodulatory, anti‐inflammatory and procoagulant properties of protein no. 4 secreted from the rat seminal vesicle epithelium (SV‐IV) have previously been found to be modulated by a supramolecular monomer–trimer equilibrium. More structural details that integrate experimental data into a predictive framework have recently been reported. Unfortunately, homology modelling and fold‐recognition strategies were not successful in creating a theoretical model of the structural organization of SV‐IV. It was inferred that the global structure of SV‐IV is not similar to that of any protein of known three‐dimensional structure. Reversing the classical approach to the sequence–structure–function paradigm, in this paper we report novel information obtained by comparing the physicochemical parameters of SV‐IV with two datasets composed of intrinsically unfolded and ideally globular proteins. In addition, we analyse the SV‐IV sequence by several publicly available disorder‐oriented predictors. Overall, disorder predictions and a re‐examination of existing experimental data strongly suggest that SV‐IV needs large plasticity to efficiently interact with the different targets that characterize its multifaceted biological function, and should therefore be better classified as an intrinsically disordered protein.
Heterologous expression of a serine carboxypeptidase‐like acyltransferase and characterization of the kinetic mechanism
Authors: Stehle Felix , Stubbs Milton T. , Strack Dieter , Milkowski Carsten
Published: January 2008

In plant secondary metabolism, β‐acetal ester‐dependent acyltransferases, such as the 1‐O‐sinapoyl‐β‐glucose:l‐malate sinapoyltransferase (SMT; EC 2.3.1.92), are homologous to serine carboxypeptidases. Mutant analyses and modeling of Arabidopsis SMT (AtSMT) have predicted amino acid residues involved in substrate recognition and catalysis, confirming the main functional elements conserved within the serine carboxypeptidase protein family. However, the functional shift from hydrolytic to acyltransferase activity and structure–function relationship of AtSMT remain obscure. To address these questions, a heterologous expression system for AtSMT has been developed that relies on Saccharomyces cerevisiae and an episomal leu2‐d vector. Codon usage adaptation of AtSMT cDNA raised the produced SMT activity by a factor of approximately three. N‐terminal fusion to the leader peptide from yeast proteinase A and transfer of this expression cassette to a high copy vector led to further increase in SMT expression by factors of 12 and 42, respectively. Finally, upscaling the biomass production by fermenter cultivation lead to another 90‐fold increase, resulting in an overall 3900‐fold activity compared to the AtSMT cDNA of plant origin. Detailed kinetic analyses of the recombinant protein indicated a random sequential bi‐bi mechanism for the SMT‐catalyzed transacylation, in contrast to a double displacement (ping‐pong) mechanism, characteristic of serine carboxypeptidases.
Emergency medicine in Sri Lanka: The inevitable evolution to a new specialty
Authors: Tan Gim , Goonasekera Chula , Curran Shane
Published: January 2008

Sri Lanka is a low‐income country with a relatively advanced, equitable and accessible health‐care system offered to its 20 million populations free of charge through a national pro‐poor health policy. Its weaknesses in emergency services, however, surfaced in 2004 when it faced the Tsunami, the worst natural disaster of the world of the 21st century. Since then, the local health community with the assistance of the government and foreign aid agencies have embarked on a path to establish emergency services, improve its preparedness for disaster management and establishment of emergency medicine training. The present article traces this path and how it is evolving in the country.
First aid for jellyfish stings: Do we really know what we are doing?
Authors: Little Mark
Published: January 2008

Jellyfish stings remains a common envenoming, and yet confusion appears to exist in the community as to the correct first aid. Current guidelines from the Australian Resuscitation Council still recommends ice for most jellyfish stiings, although there appears to belittle evidence to support this. There is more evidence supporting the use of hot water. More research is required to simplify first aid for jellyfish stings.
The spirit of capitalism, stock market bubbles and output fluctuations
Authors: Kamihigashi Takashi
Published: January 2008

This paper presents a representative agent model in which stock market bubbles cause output fluctuations. Assuming that utility depends directly on wealth, we show that stock market bubbles arise if the marginal utility of wealth does not decline to zero as wealth goes to infinity. Bubbles can affect output positively or negative depending on whether the production function exhibits increasing or decreasing returns to scale. In sunspot equilibria, the bursting of a bubble is followed by a sharp decline in output one period later. Various numerical examples are given to illustrate the behavior of stochastic bubbles and the relationship between bubbles and output.
Time preference and two‐country trade
Authors: Chen Been‐Lon , Nishimura Kazuo , Shimomura Koji
Published: January 2008

We present a dynamic two‐country model of international trade with endogenous time preference. We show that if the two countries have similar preferences, production technologies and labor endowments, there exists a unique and stable steady state such that both consumption and investment goods are produced in both countries. Unlike the case of constant time preferences, the steady state is independent of the initial international distribution of capital. We prove a dynamic Heckscher–Ohlin theorem such that the labor‐abundant country exports the labor‐intensive good.
Altruism as a motive for intergenerational transfers
Authors: Fujiu Hiroshi , Yano Makoto
Published: January 2008

This study investigates the role of altruism as a motive for transfer payments. In the existing literature on altruism, it is generally assumed that a transfer payment is made out of altruism that a transferor feels towards a transferee. This study demonstrates that this is not necessarily the case. By using a dynamic model in which children are altruistic towards parents, it demonstrates that such altruism may induce a parent to give a transfer to children.
Chemosensitivity Determinants of Irinotecan Hydrochloride in Hepatocellular Carcinoma Cell Lines
Authors: Takahata Takenori , Ookawa Keizou , Suto Koji , Tanaka Masanori , Yano Hirohisa , Nakashima Osamu , Kojiro Masamichi , Tamura Yoshihiro , Tateishi Tomonori , Sakata Yuh , Fukuda Shinsaku
Published: January 2008

Abstract:  Irinotecan hydrochloride (CPT‐11) is an effective anticancer drug, and its metabolic pathway has been well studied. Nevertheless, in human hepatocellular carcinoma (HCC), its cytotoxicity is less well studied and the determination of its chemosensitivity is unclear. We, therefore, examined chemosensitivities of HCC cell lines for CPT‐11 and SN‐38, and mRNA expressions of several molecules in their metabolic pathway. Three markers were found to correlate well with chemosensitivity: breast cancer resistance protein (BCRP), cytochrome P450 (CYP) 3A4 and uridine diphosphate‐glucuronosyl transferase (UGT) 1A1. Next, CYP3A4/UGT1A1/BCRP inhibitor naringenin and BCRP inhibitor elacridar were tested for enhancement of their chemosensitivity. Both of naringenin and elacridar separately enhanced the sensitivity for CPT‐11 and SN‐38 in KYN‐2 cells abundantly expressing BCRP, CYP3A4/5 and UGT1A1, but not in KYN‐1 cells expressing lower levels. However, naringenin had little effect on the sensitivity in JHH‐4 and HLE cells with higher CYP3A4/5 and lower UGT1A1 and BCRP expression. On the other hand, naringenin and elacridar significantly increased the chemosensitivity for CPT‐11 and SN‐38 in the KYN‐1‐derived cells artificially overexpressing BCRP. Furthermore, flow cytometric analysis showed that naringenin raised intracellular accumulation of CPT‐11 as well as elacridar. Those results suggest that BCRP is one of the chemosensitivity determinants of CPT‐11 in HCC cells and its inhibition might be critical for cells expressing abundant BCRP.