bluuebunny/crossref_metadata_2025 huggingface dataset:
The establishment and characterization of the first canine hepatocellular carcinoma cell line, which resembles human oncogenic expression patterns
Authors: Boomkens Sacha Y , Spee Bart , IJzer Jooske , Kisjes Ronald , Egberink Herman F , van den Ingh Ted SGAM , Rothuizen Jan , Penning Louis C
Published: January 2005

Background

Hepatocellular carcinoma (HCC) is one of the most worldwide frequent primary carcinomas resulting in the death of many cirrhotic patients. Unfortunately, the molecular mechanisms of this cancer are not well understood; therefore, we need a good model system to study HCC. The dog is recognized as a promising model for human medical research, namely compared with rodents. The objective of this study was to establish and characterize a spontaneous canine tumor cell line as a potential model for studies on HCC.

Results

Histomorphological, biochemical, molecular biological and quantitative assays were performed to characterize the canine HCC cell line that originated from a dog with a spontaneous liver tumor. Morphological investigations provided strong evidence for the hepatocytic and neoplastic nature of the cell line, while biochemical assays showed that they produced liver-specific enzymes. PCR analysis confirmed expression of ceruloplasmin, alpha-fetoprotein and serum albumin. Quantitative RT-PCR showed that the canine HCC cell line resembles human HCC based on the measurements of expression profiles of genes involved in cell proliferation and apoptosis.

Conclusions

We have developed a novel, spontaneous tumor liver cell line of canine origin that has many characteristics of human HCC. Therefore, the canine HCC cell line might be an excellent model for comparative studies on the molecular pathogenesis of HCC.
Reconstruction of the pelvic ring using an autologous free non-vascularized fibula graft in a patient with benign fibrous histiocytoma
Authors: Niemeyer Philipp , Ludwig Karl , Werner Mathias , Bernd Ludger , Parsch Dominik
Published: January 2005

Background

Benign fibrous histiocytomas (BFH) usually presents as a small benign lesion that predominantly occurs in the skin. Only few cases of BFH arising from bone have been reported, its occurrence in pelvic bones is even rarer.

Case presentation

A 34-year-old female presented with BFH at a rare anatomical location on both sides of the os ilium which was larger than earlier reported BFH of the bone. Surgical resection was performed successfully including resection of the inner pelvic ring and reconstruction of the linea terminalis using a non-vascularized fibular autograft. At 18 months after tumor resection and reconstruction of the pelvic ring, with interposition of a free vascularized fibula graft patient has an excellent clinical oncological and functional outcome.

Conclusion

Non vascularized fibular autograft is a useful reconstructive procedure in select patients.
Odontogenic tumors in Nigerian children and adolescents- a retrospective study of 92 cases
Authors: Ajayi Oluseyi F , Ladeinde Akinola L , Adeyemo Wasiu L , Ogunlewe Mobolanle O
Published: January 2005

Background

Tumours arising from odontogenic tissues are rare and constitute a heterogenous group of interesting lesions. The aim of this study was to determine the relative frequency of odontogenic tumors (OT) among Nigerian children and adolescents 19 years or younger.

Patients and methods

The histopathology records were retrospectively reviewed for all the tumors and tumor-like lesions of the oral cavity and the jaws seen in children and adolescents ≤ 19 years seen between January 1980 and December 2003. Hematoxylin and eosin-stained sections were re-evaluated and the diagnosis in each case was confirmed or modified according to World Health Organization (WHO) classification, 1992; and were subjected to analysis of age, sex, site of tumor and histopathologic type.

Results

A total of 477 tumors and tumor-like lesions were seen in patients ≤ 19 years during the period of the study. Of these, 92 (19.3%) were odontogenic tumors. Benign odontogenic tumors constituted 98.9% of the cases seen, while only 1 case (1.1%) of malignant variety was seen during the period. The mean (SD) age of patients was 14.9 (± 3.1) years (range, 4–19 years). Male-to-female ratio was 1:1; and mandible-to-maxilla ratio was 2.7:1. OT's were most frequently seen in patients aged 16–19 years (46.7%) and the least number (2.2%) were found in patients aged 0–5 years. Among nine histologic types of OT seen, ameloblastoma (48.9%), adenomatoid odontogenic tumor (19.6%) and odontogenic myxoma (8.7%) were predominant. Multicystic/solid and unicystic variants of ameloblastoma were diagnosed in 40 (89%) and 5 (11%) cases respectively.

Conclusions

Odontogenic tumors are relatively common in children and adolescents in Nigeria. One out of every 5 children and adolescents with tumors and tumor-like lesions of oral cavity and the jaws seen in this study had a diagnosis of odontogenic tumor.
Ectopic paraesophageal mediastinal parathyroid adenoma, a rare cause of acute pancreatitis
Authors: Foroulis Christophoros N , Rousogiannis Sotirios , Lioupis Christos , Koutarelos Dimitrios , Kassi Georgia , Lioupis Athanassios
Published: January 2005

Background

The manifestation of primary hyperparathyroidism with acute pancreatitis is a rare event. Ectopic paraesophageal parathyroid adenomas account for about 5%–10% of primary hyperparathyroidism and surgical resection results in cure of the disease.

Case presentation

A 71-year-old woman was presented with acute pancreatitis and hypercalcaemia. During the investigation of hypercalcemia, a paraesophageal ectopic parathyroid mass was detected by computerized tomography (CT) scan and 99mTc sestamibi scintigraphy. The tumor was resected via a cervical collar incision and calcium and parathormone tumor levels returned to normal within 48 hours.

Conclusions

Acute pancreatitis associated with hypercalcaemia should pose the suspicion of primary hyperparathyroidism. Accurate preoperative localization of an ectopic parathyroid adenoma, by using the combination of 99mTc sestamibi scintigraphy and CT scan of the neck and chest allows successful surgical treatment.
Open Access to essential health care information
Authors: Stokes Christabel EL , Pandey Manoj
Published: January 2005

Open Access publishing is a valuable resource for the synthesis and distribution of essential health care information. This article discusses the potential benefits of Open Access, specifically in terms of Low and Middle Income (LAMI) countries in which there is currently a lack of informed health care providers – mainly a consequence of poor availability to information. We propose that without copyright restrictions, Open Access facilitates distribution of the most relevant research and health care information. Furthermore, we suggest that the technology and infrastructure that has been put in place for Open Access could be used to publish download-able manuals, guides or basic handbooks created by healthcare providers in LAMI countries.
Public sector reform and demand for human resources for health (HRH)
Authors: Lethbridge Jane
Published: January 2005

This article considers some of the effects of health sector reform on human resources for health (HRH) in developing countries and countries in transition by examining the effect of fiscal reform and the introduction of decentralisation and market mechanisms to the health sector.

Fiscal reform results in pressure to measure the staff outputs of the health sector. Financial decentralisation often leads to hospitals becoming "corporatised" institutions, operating with business principles but remaining in the public sector. The introduction of market mechanisms often involves the formation of an internal market within the health sector and market testing of different functions with the private sector. This has immediate implications for the employment of health workers in the public sector, because the public sector may reduce its workforce if services are purchased from other sectors or may introduce more short-term and temporary employment contracts.

Decentralisation of budgets and administrative functions can affect the health sector, often in negative ways, by reducing resources available and confusing lines of accountability for health workers. Governance and regulation of health care, when delivered by both public and private providers, require new systems of regulation.

The increase in private sector provision has led health workers to move to the private sector. For those remaining in the public sector, there are often worsening working conditions, a lack of employment security and dismantling of collective bargaining agreements.

Human resource development is gradually being recognised as crucial to future reforms and the formulation of health policy. New information systems at local and regional level will be needed to collect data on human resources. New employment arrangements, strengthening organisational culture, training and continuing education will also be needed.
Transfer of Health for All policy – What, how and in which direction? A two-case study
Authors: Tervonen-Gonçalves Leena , Lehto Juhani
Published: January 2005

Background

This article explores the transfer of World Health Organization's (WHO) policy initiative Health for All by the Year 2000 (HFA2000) into national contexts by using the changes in the public health policies of Finland and Portugal from the 1970's onward and the relationship of these changes to WHO policy development as test cases. Finland and Portugal were chosen to be compared as they represent different welfare state types and as the paradigmatic transition from the old to new public health is assumed to be related to the wider welfare state development.

Methods

The policy transfer approach is used as a conceptual tool to analyze the possible policy changes related to the adaptation of HFA into the national context. To be able to analyze not only the content but also the contextual conditions of policy transfer Kingdon's analytical framework of policy analysis is applied.

Conclusions

Our analysis suggests that no significant change of health promotion policy resulted from the launch of HFA program neither in Finland nor in Portugal. Instead the changes that occurred in both countries were of incremental nature, in accordance with the earlier policy choices, and the adaptation of HFA program was mainly applied to the areas where there were national traditions.
RNA interference: learning gene knock-down from cell physiology
Authors: Mocellin Simone , Provenzano Maurizio
Published: January 2005

Over the past decade RNA interference (RNAi) has emerged as a natural mechanism for silencing gene expression. This ancient cellular antiviral response can be exploited to allow specific inhibition of the function of any chosen target gene. RNAi is proving to be an invaluable research tool, allowing much more rapid characterization of the function of known genes. More importantly, RNAi technology considerably bolsters functional genomics to aid in the identification of novel genes involved in disease processes.

This review briefly describes the molecular principles underlying the biology of RNAi phenomenon and discuss the main technical issues regarding optimization of RNAi experimental design.
Vaccination with EphA2-derived T cell-epitopes promotes immunity against both EphA2-expressing and EphA2-negative tumors
Authors: Hatano Manabu , Kuwashima Naruo , Tatsumi Tomohide , Dusak Jill E , Nishimura Fumihiko , Reilly Karlyne M , Storkus Walter J , Okada Hideho
Published: January 2005

Background

A novel tyrosine kinase receptor EphA2 is expressed at high levels in advanced and metastatic cancers. We examined whether vaccinations with synthetic mouse EphA2 (mEphA2)-derived peptides that serve as T cell epitopes could induce protective and therapeutic anti-tumor immunity.

Methods

C57BL/6 mice received subcutaneous (s.c.) vaccinations with bone marrow-derived dendritic cells (DCs) pulsed with synthetic peptides recognized by CD8+ (mEphA2671–679, mEphA2682–689) and CD4+ (mEphA230–44) T cells. Splenocytes (SPCs) were harvested from primed mice to assess the induction of cytotoxic T lymphocyte (CTL) responses against syngeneic glioma, sarcoma and melanoma cell lines. The ability of these vaccines to prevent or treat tumor (s.c. injected MCA205 sarcoma or B16 melanoma; i.v. injected B16-BL6) establishment/progression was then assessed.

Results

Immunization of C57BL/6 mice with mEphA2-derived peptides induced specific CTL responses in SPCs. Vaccination with mEPhA2 peptides, but not control ovalbumin (OVA) peptides, prevented the establishment or prevented the growth of EphA2+ or EphA2-negative syngeneic tumors in both s.c. and lung metastasis models.

Conclusions

These data indicate that mEphA2 can serve as an attractive target against which to direct anti-tumor immunity. The ability of mEphA2 vaccines to impact EphA2-negative tumors such as the B16 melanoma may suggest that such beneficial immunity may be directed against alternative EphA2+ target cells, such as the tumor-associated vascular endothelial cells.
Search for computational modules in the C. elegansbrain
Authors: Reigl Markus , Alon Uri , Chklovskii Dmitri B
Published: January 2005

Background

Does the C. elegans nervous system contain multi-neuron computational modules that perform stereotypical functions? We attempt to answer this question by searching for recurring multi-neuron inter-connectivity patterns in the C. elegans nervous system's wiring diagram.

Results

Our statistical analysis reveals that some inter-connectivity patterns containing two, three and four (but not five) neurons are significantly over-represented relative to the expectations based on the statistics of smaller inter-connectivity patterns.

Conclusions

Over-represented patterns (or motifs) are candidates for computational modules that may perform stereotypical functions in the C. elegans nervous system. These modules may appear in other species and need to be investigated further.
HTLV-1 and -2 envelope SU subdomains and critical determinants in receptor binding
Authors: Kim Felix J , Manel Nicolas , Garrido Edith N , Valle Carine , Sitbon Marc , Battini Jean-Luc
Published: January 2005

Background

Human T-cell leukemia virus (HTLV) -1 and -2 are deltaretroviruses that infect a wide range of cells. Glut1, the major vertebrate glucose transporter, has been shown to be the HTLV Env receptor. While it is well established that the extracellular surface component (SU) of the HTLV envelope glycoprotein (Env) harbors all of the determinants of interaction with the receptor, identification of SU subdomains that are necessary and sufficient for interaction with the receptor, as well as critical amino acids therein, remain to be precisely defined. Although highly divergent in the rest of their genomes, HTLV and murine leukemia virus (MLV) Env appear to be related and based on homologous motifs between the HTLV and MLV SU, we derived chimeric HTLV/MLV Env and soluble HTLV-1 and -2 truncated amino terminal SU subdomains.

Results

Using these SU constructs, we found that the 183 and 178 amino terminal residues of the HTLV-1 and -2 Env, respectively, were sufficient to efficiently bind target cells of different species. Binding resulted from bona fide interaction with the HTLV receptor as isolated SU subdomains specifically interfered with HTLV Env-mediated binding, cell fusion, and cell-free as well as cell-to-cell infection. Therefore, the HTLV receptor-binding domain (RBD) lies in the amino terminus of the SU, immediately upstream of a central immunodominant proline rich region (Env residues 180 to 205), that we show to be dispensible for receptor-binding and interference. Moreover, we identified a highly conserved tyrosine residue at position 114 of HTLV-1 Env, Tyr114, as critical for receptor-binding and subsequent interference to cell-to-cell fusion and infection. Finally, we observed that residues in the vicinity of Tyr114 have lesser impact on receptor binding and had various efficiency in interference to post-binding events.

Conclusions

The first 160 residues of the HTLV-1 and -2 mature cleaved SU fold as autonomous domains that contain all the determinants required for binding the HTLV receptor.
Alterations in the expression of DEAD-box and other RNA binding proteins during HIV-1 replication
Authors: Krishnan Vyjayanthi , Zeichner Steven L
Published: January 2005

Recent results showed that certain DEAD box protein RNA helicases, DDX3 and DDX1, play an important role in the HIV infection cycle by facilitating the export of long, singly spliced or unspliced HIV RNAs from the nucleus via the CRM1-Rev pathway. Close examination of an extensive microarray expression profiling dataset obtained from cells latently infected with HIV induced to undergo lytic viral replication indicated that additional DEAD box proteins, beyond DDX3 and DDX1, exhibit differential expression during lytic HIV replication, and in latently infected cells prior to induction into active replication. This finding provides additional evidence that the involvement of DEAD box proteins and other RNA-binding proteins may play roles in active HIV replication and in the control of viral latency. Agents targeting these functions may offer new approaches to antiretroviral therapy and the therapeutic manipulation of HIV latency.
Homologs of wingless and decapentaplegic display a complex and dynamic expression profile during appendage development in the millipede Glomeris marginata (Myriapoda: Diplopoda)
Authors: Prpic Nikola-Michael
Published: January 2005

Background

The Drosophila genes wingless (wg) and decapentaplegic (dpp) comprise the top level of a hierarchical gene cascade involved in proximal-distal (PD) patterning of the legs. It remains unclear, whether this cascade is common to the appendages of all arthropods. Here, wg and dpp are studied in the millipede Glomeris marginata, a representative of the Myriapoda.

Results

Glomeris wg (Gm-wg) is expressed along the ventral side of the appendages compatible with functioning during the patterning of both the PD and dorsal-ventral (DV) axes. Gm-wg may also be involved in sensory organ formation in the gnathal appendages by inducing the expression of Distal-less (Dll) and H15 in the organ primordia. Expression of Glomeris dpp (Gm-dpp) is found at the tip of the trunk legs as well as weakly along the dorsal side of the legs in early stages. Taking data from other arthropods into account, these results may be interpreted in favor of a conserved mode of WG/DPP signaling. Apart from the main PD axis, many arthropod appendages have additional branches (e.g. endites). It is debated whether these extra branches develop their PD axis via the same mechanism as the main PD axis, or whether branch-specific mechanisms exist. Gene expression in possible endite homologs in Glomeris argues for the latter alternative.

Conclusion

All available data argue in favor of a conserved role of WG/DPP morphogen gradients in guiding the development of the main PD axis. Additional branches in multibranched (multiramous) appendage types apparently do not utilize the WG/DPP signaling system for their PD development. This further supports recent work on crustaceans and insects, that lead to similar conclusions.
Reaching in reality and virtual reality: a comparison of movement kinematics in healthy subjects and in adults with hemiparesis
Authors: Viau Antonin , Feldman Anatol G , McFadyen Bradford J , Levin Mindy F
Published: January 2005

Background

Virtual reality (VR) is an innovative tool for sensorimotor rehabilitation increasingly being employed in clinical and community settings. Despite the growing interest in VR, few studies have determined the validity of movements made in VR environments with respect to real physical environments. The goal of this study was to compare movements done in physical and virtual environments in adults with motor deficits to those in healthy individuals.

Methods

The participants were 8 healthy adults and 7 adults with mild left hemiparesis due to stroke. Kinematics of functional arm movements involving reaching, grasping and releasing made in physical and virtual environments were analyzed in two phases: 1) reaching and grasping the ball and 2) ball transport and release. The virtual environment included interaction with an object on a 2D computer screen and haptic force feedback from a virtual ball. Temporal and spatial parameters of reaching and grasping were determined for each phase.

Results

Individuals in both groups were able to reach, grasp, transport, place and release the virtual and real ball using similar movement strategies. In healthy subjects, reaching and grasping movements in both environments were similar but these subjects used less wrist extension and more elbow extension to place the ball on the virtual vertical surface. Participants with hemiparesis made slower movements in both environments compared to healthy subjects and during transport and placing of the ball, trajectories were more curved and interjoint coordination was altered. Despite these differences, patients with hemiparesis also tended to use less wrist extension during the whole movement and more elbow extension at the end of the placing phase.

Conclusion

Differences in movements made by healthy subjects in the two environments may be explained by the use of a 2D instead of a 3D virtual environment and the absence of haptic feedback from the VR target. Despite these differences, our findings suggest that both healthy subjects and individuals with motor deficits used similar movement strategies when grasping and placing a ball in the two reality conditions. This suggests that training of arm movements in VR environments may be a valid approach to the rehabilitation of patients with motor disorders.
Susceptibility of different leukocyte cell types to Vaccinia virus infection
Authors: Sánchez-Puig Juana M , Sánchez Laura , Roy Garbiñe , Blasco Rafael
Published: January 2005

Background

Vaccinia virus, the prototype member of the family Poxviridae, was used extensively in the past as the Smallpox vaccine, and is currently considered as a candidate vector for new recombinant vaccines. Vaccinia virus has a wide host range, and is known to infect cultures of a variety of cell lines of mammalian origin. However, little is known about the virus tropism in human leukocyte populations. We report here that various cell types within leukocyte populations have widely different susceptibility to infection with vaccinia virus.

Results

We have investigated the ability of vaccinia virus to infect human PBLs by using virus recombinants expressing green fluorescent protein (GFP), and monoclonal antibodies specific for PBL subpopulations. Flow cytometry allowed the identification of infected cells within the PBL mixture 1–5 hours after infection. Antibody labeling revealed that different cell populations had very different infection rates. Monocytes showed the highest percentage of infected cells, followed by B lymphocytes and NK cells. In contrast to those cell types, the rate of infection of T lymphocytes was low. Comparison of vaccinia virus strains WR and MVA showed that both strains infected efficiently the monocyte population, although producing different expression levels. Our results suggest that MVA was less efficient than WR in infecting NK cells and B lymphocytes. Overall, both WR and MVA consistently showed a strong preference for the infection of non-T cells.

Conclusions

When infecting fresh human PBL preparations, vaccinia virus showed a strong bias towards the infection of monocytes, followed by B lymphocytes and NK cells. In contrast, very poor infection of T lymphocytes was detected. These finding may have important implications both in our understanding of poxvirus pathogenesis and in the development of improved smallpox vaccines.
Thermodynamics of weight loss diets
Authors: Fine Eugene J , Feinman Richard D
Published: January 2005

Background

It is commonly held that "a calorie is a calorie", i.e. that diets of equal caloric content will result in identical weight change independent of macronutrient composition, and appeal is frequently made to the laws of thermodynamics. We have previously shown that thermodynamics does not support such a view and that diets of different macronutrient content may be expected to induce different changes in body mass. Low carbohydrate diets in particular have claimed a "metabolic advantage" meaning more weight loss than in isocaloric diets of higher carbohydrate content. In this review, for pedagogic clarity, we reframe the theoretical discussion to directly link thermodynamic inefficiency to weight change. The problem in outline: Is metabolic advantage theoretically possible? If so, what biochemical mechanisms might plausibly explain it? Finally, what experimental evidence exists to determine whether it does or does not occur?

Results

Reduced thermodynamic efficiency will result in increased weight loss. The laws of thermodynamics are silent on the existence of variable thermodynamic efficiency in metabolic processes. Therefore such variability is permitted and can be related to differences in weight lost. The existence of variable efficiency and metabolic advantage is therefore an empiric question rather than a theoretical one, confirmed by many experimental isocaloric studies, pending a properly performed meta-analysis. Mechanisms are as yet unknown, but plausible mechanisms at the metabolic level are proposed.

Conclusions

Variable thermodynamic efficiency due to dietary manipulation is permitted by physical laws, is supported by much experimental data, and may be reasonably explained by plausible mechanisms.
CXCR3/CXCL10 expression in the synovium of children with juvenile idiopathic arthritis
Authors: Martini Georgia , Zulian Francesco , Calabrese Fiorella , Bortoli Marta , Facco Monica , Cabrelle Anna , Valente Marialuisa , Zacchello Franco , Agostini Carlo
Published: January 2005

The accumulation of T cells in the synovial membrane is the crucial step in the pathophysiology of the inflammatory processes characterizing juvenile idiopathic arthritis (JIA). In this study, we evaluated the expression and the pathogenetic role in oligoarticular JIA of a CXC chemokine involved in the directional migration of activated T cells, i.e. IFNγ-inducible protein 10 (CXCL10) and its receptor, CXCR3. Immunochemistry with an antihuman CXCL10 showed that synovial macrophages, epithelial cells, and endothelial cells bear the chemokine. By flow cytometry and immunochemistry, it has been shown that CXCR3 is expressed at high density by virtually all T lymphocytes isolated from synovial fluid (SF) and infiltrating the synovial membrane. Particularly strongly stained CXCR3+ T cells can be observed close to the luminal space and in the perivascular area. Furthermore, densitometric analysis has revealed that the mRNA levels for CXCR3 are significantly higher in JIA patients than in controls. T cells purified from SF exhibit a definite migratory capability in response to CXCL10. Furthermore, SF exerts significant chemotactic activity on the CXCR3+ T-cell line, and this activity is inhibited by the addition of an anti-CXCL10 neutralizing antibody. Taken together, these data suggest that CXCR3/CXCL10 interactions are involved in the pathophysiology of JIA-associated inflammatory processes, regulating both the activation of T cells and their recruitment into the inflamed synovium.
Autoantibodies specific for apoptotic U1-70K are superior serological markers for mixed connective tissue disease
Authors: Hof Daniëlle , Cheung Kalok , de Rooij Dirk-Jan RAM , van den Hoogen Frank H , Pruijn Ger JM , van Venrooij Walther J , Raats Jos MH
Published: January 2005

Modifications occurring on autoantigens during cell death have been proposed to have a role in the initiation of autoimmune diseases. Patients suffering from mixed connective tissue disease (MCTD) produce autoantibodies directed to U1 small nuclear ribonucleoprotein (snRNP), and antibodies against a 70 kDa protein component, the U1-70K (70K) protein, are the most prominent. During apoptosis, 70K is cleaved by caspase-3 to a 40 kDa product, which remains associated with the complex. Autoantibodies preferentially recognizing the apoptotic form of 70K have been described previously, and an apoptosis-specific epitope on 70K has been identified. This study shows that 29 of 53 (54%) MCTD sera preferentially recognize the apoptotic form of 70K over intact 70K. Moreover, we show that antibodies directed to an apoptosis-specific epitope on 70K are more specifically associated with MCTD than other anti-70K antibodies, suggesting that apoptotic 70K is a better antigen for the detection of these antibodies in MCTD patients. Longitudinal analysis of 12 MCTD patients showed in several patients that early sera are relatively enriched with antibodies recognizing an apoptosis-specific epitope, and that the levels of these apoptosis-specific antibodies decrease in time. These findings indicate that the early detection of apoptotic 70K is of considerable interest for anti-U1 snRNP-positive patients.
Full-length cDNAs from chicken bursal lymphocytes to facilitate gene function analysis
Authors: Caldwell Randolph B , Kierzek Andrzej M , Arakawa Hiroshi , Bezzubov Yuri , Zaim Jolanta , Fiedler Petra , Kutter Stefan , Blagodatski Artem , Kostovska Diyana , Koter Marek , Plachy Jiri , Carninci Piero , Hayashizaki Yoshihide , Buerstedde Jean-Marie
Published: January 2005

A large number of cDNA inserts were sequenced from a high-quality library of chicken bursal lymphocyte cDNAs. Comparisons to public gene databases indicate that the cDNA collection represents more than 2,000 new, full-length transcripts. This resource defines the structure and the coding potential of a large fraction of B-cell specific and housekeeping genes whose function can be analyzed by disruption in the chicken DT40 B-cell line.
Activated human PMN synthesize and release a strongly fucosylated glycoform of α1-acid glycoprotein, which is transiently deposited in human myocardial infarction
Authors: Poland Dennis C W , Vallejo Juan-Jesús García , Niessen Hans W M , Nijmeyer Remco , Calafat Jero , Hack C Erik , Van het Hof Bert , Van Dijk Willem
Published: January 2005

α1-Acid glycoprotein (AGP) is a major acute-phase protein present in human plasma as well as in polymorphonuclear leukocytes (PMN). In this report, we show that PMN synthesize a specific glycoform of AGP, which is stored in the specific and azurophilic granules. Activation of PMN results in the rapid release of soluble AGP. PMN AGP exhibits a substantially higher apparent molecular weight than plasma AGP (50–60 kD vs. 40–43 kD), owing to the presence of strongly fucosylated and sialylated polylactosamine units on its five N-linked glycans. PMN AGP is also released in vivo from activated PMN, as appeared from studies using well-characterized myocard slices of patients that had died within 2 weeks after an acute myocardial infarction. AGP was found deposited transiently on damaged cardiomyocytes in areas with infiltrating PMN only. It is interesting that this was inversely related to the deposition of activated complement C3. Strongly fucosylated and sialylated AGP glycoforms have the ability to bind to E-selectin and to inhibit complement activation. We suggest that AGP glycoforms in PMN provide an endogenous feedback-inhibitory response to excessive inflammation.
Molecular Mechanisms of Fenretinide‐Induced Apoptosis of Neuroblastoma Cells
Authors: LOVAT PENNY E. , CORAZZARI MARCO , GORANOV BOJIDAR , PIACENTINI MAURO , REDFERN CHRISTOPHER P. F.
Published: January 2005

Abstract: Synthetic retinoids such as fenretinide [N‐(4‐hydroxyphenyl)retinamide] induce apoptosis of neuroblastoma cells, act synergistically with chemotherapeutic drugs, and may provide opportunities for novel approaches to neuroblastoma therapy. Fenretinide‐induced cell death of neuroblastoma cells is caspase dependent and results in the release of cytochrome c from mitochondria independently of changes in permeability transition. This is mediated by a signaling pathway characterized by the generation of reactive oxygen species (ROS) via 12‐lipoxygenase (12‐LOX), and an oxidative‐stress‐dependent induction of the transcription factor, GADD153 and the BCL2‐related protein BAK. Upstream events of fenretinide‐induced signaling involve increased levels of ceramide as a result of increased sphingomyelinase activity, and the subsequent metabolism of ceramide to gangliosides via glucosylceramide synthase and GD3 synthase. These gangliosides may be involved in the regulation of 12‐LOX leading to oxidative stress and apoptosis via the induction of GADD153 and BAK. The targeting of sphingomyelinases or downstream effectors such as 12‐LOX or GADD153 may present novel approaches for the development of more effective and selective drugs for neuroblastoma therapy.
DNA Minigene Vaccination for Adjuvant Neuroblastoma Therapy
Authors: LODE HOLGER N. , HUEBENER NICOLE , ZENG YAN , FEST STEFAN , WEIXLER S , GAEDICKE GERHARD
Published: January 2005

Abstract: The disruption of self‐tolerance against neuroblastoma is the ultimate goal of an effective DNA‐vaccine. We demonstrate the induction of protective immunity against syngeneic murine NXS2 neuroblastoma in A/J mice following vaccination with tyrosine hydroxylase (TH)‐derived antigens. Oral gene delivery was accomplished using an attenuated strain of Salmonella typhimurium as a carrier harboring vectors encoding for mouse tyrosine hydroxylase (mTH) antigens. Vaccination was effective in protecting animals from a lethal challenge with wild‐type NXS2 tumor cells. These findings were extended by comparing efficacy of mTH minigene vaccines with a minigene vaccine comprising three novel epitopes isolated fom NXS2 neuroblastoma cells. For this purpose, MHC class I was immunoprecipitated from NXS2 cell lysates, and peptides were eluted and examined in tandem‐mass spectrometry analysis. This led to the identification of three novel natural MHC class I peptide ligands: TEALPVKLI, from ribonucleotide reductase M2; NEYIMSLI, from Ser/Thr protein phosphatase 2A; and FEMVSTLI, of unknown origin. Two minigenes were constructed, one encoding for the three novel epitopes and the second for three known mTH‐derived epitopes with high predicted binding affinity to MHC class I, by cloning them into the mammalian expression vector pCMV‐3FUB. Immunized mice showed a reduction in primary tumor growth and the absence of spontaneous liver metastasis in the majority of animals. Importantly, there was no significant difference between the two minigenes, suggesting that, compared with tumor peptide isolation, mTH epitope prediction is similarly effective for designing efficient DNA‐minigene vaccines. In summary, these findings establish proof of the concept that disruption of self‐tolerance against neuroblastoma‐associated epitopes may be an effective adjuvant therapeutic strategy.
Apoptosis Signaling in Tumor Therapy
Authors: FULDA SIMONE , DEBATIN KLAUS‐MICHAEL
Published: January 2005

Abstract: Apoptosis, the cell's intrinsic program to death, plays a central role in physiological growth control and regulation of tissue homeostasis. Tipping the balance between cell death and proliferation in favor of cell survival may result in tumor formation. Also, killing of cancer cells by cytotoxic therapies currently used for treatment of cancer, for example, chemotherapy, γ‐irradiation, immunotherapy, or suicide gene therapy, largely depends on activation of apoptosis programs in cancer cells. Accordingly, failure to undergo apoptosis in response to anticancer therapy may result in cancer resistance. Further insights into the mechanisms regulating apoptosis in response to anticancer therapy and how cancer cells evade cell death may provide novel opportunities for drug development.
Autocrine and Paracrine Calcium Signaling by the CD38/NAD+/Cyclic ADP‐Ribose System
Authors: DE FLORA ANTONIO , ZOCCHI ELENA , GUIDA LUCREZIA , FRANCO LUISA , BRUZZONE SANTINA
Published: January 2005

Abstract: CD38, a multifunctional enzyme, generates two potent Ca2+‐releasing signal metabolites, cyclic ADP‐ribose (cADPR) and NAADP+, thereby upmodulating many important Ca2+‐mediated cell functions. A topological paradox has long been recognized, as CD38 is an ectoenzyme, or an intravesicularly located enzyme in subcellular membrane vesicles, therefore apparently shielded from its substrate NAD+. Moreover, cADPR generated by CD38 should be unavailable to its target Ca2+ stores, the ryanodine receptors (RyR). We have solved this paradox by identifying some NAD+ and cADPR transmembrane transporters, whose interplay mediates a hitherto‐unrecognized subcellular and intercellular trafficking of nucleotides that enhances intracellular Ca2+ ([Ca2+]i). Connexin 43 (Cx43) hemichannels mediate an equilibrative transport of NAD+ from the cytosol to the active site of CD38 (either ectocellular or intravesicular). Subsequent translocation of in situ‐generated cADPR to reach the RyR is performed, (i) by CD38 itself (concentrative) or (ii) by nucleoside transporters (NT) (one equilibrative and three concentrative). Besides this autocrine mechanism, the same transporters also mediate intercellular (paracrine) trafficking. Thus, Cx43+ and CD38+ cells can provide cADPR to neighboring RyR+ parenchymal cells and enhance their [Ca2+]i levels and Ca2+‐dependent functions accordingly. Examples of cADPR‐responsive cells via paracrine processes include (i) smooth myocytes, (ii) 3T3 murine fibroblasts, (iii) hippocampal neurons, and (iv) human hemopoietic stem cells.
Estrogen's Effects on Mitochondrial Gene Expression: Mechanisms and Potential Contributions to Estrogen Carcinogenesis
Authors: CHEN JIN‐QIANG , YAGER JAMES D.
Published: January 2005

Abstract: Estrogen receptor (ER)α and ERβ are localized in the nucleus and involved in the regulation of nuclear estrogen‐responsive genes by 17β estradiol (E2). In addition, recently others have shown that upon E2 binding, ERα localizes to the plasma membrane and initiates mitogen‐activated protein kinase (MAPK)‐mediated signal transduction. Previously, we reported that in liver, cultured rat hepatocytes and human HepG2 cells, estrogen treatment enhanced mitochondrial DNA (mtDNA)‐encoded gene transcript levels. These effects were blocked by a specific antiestrogen, suggesting a role for the ER. Others have reported the presence of putative estrogen‐responsive elements in mtDNA. These observations suggested the hypothesis that the ER localized in mitochondria and functioned directly to enhance the levels of mtDNA‐encoded transcripts, analogous to what has been observed for the glucocorticoid hormone receptor. Using Western blot analysis, confocal immunofluorescence, immunogold electron microscopy, and gel electrophoresis mobility shift assays, we have demonstrated the estrogen‐dependent presence of ERβ and ERα within mitochondria of HepG2 and MCF‐7 human breast tumor cells. Together, these results suggest that the ERs may act as transcription factors directly involved in the regulation by E2 of mtDNA transcription.
The Role of Endogenous Hormones in the Etiology and Prevention of Breast Cancer: The Epidemiological Evidence
Authors: MUTI PAOLA
Published: January 2005

Abstract: Breast cancer is the most common cause of cancer death in women worldwide. Rates vary about fivefold around the world, but they are increasing in regions that until recently had low rates of the disease. Despite the numerous uncertainties surrounding the etiology of breast cancer, intensive epidemiological, clinical, and genetic studies have identified a number of biologic and social traits as risk factors associated with it. Principal among them are the evidence of BRCA1 and BRCA2 susceptibility genes, familial history of breast cancer, age, higher socioeconomic status, ionizing radiation, tallness in adult life, alcohol consumption, and a variety of hormonal and metabolic factors. Among the hormonal influences, a relevant etiological function has been ascribed to unopposed exposure to elevated levels of estrogens and androgens. In addition, new epidemiological evidence indicates that among the metabolic factors, glucose metabolism, hyperinsulinemic insulin resistance, and insulin‐like growth factor bioavailability may also play a role in breast cancer. These endocrine and metabolic factors may represent future targets for breast cancer prevention.
A Multidisciplinary Approach to the Study of T Cell Migration
Authors: LONG AIDEEN , MITCHELL SIOBHAN , KASHANIN DMITRY , WILLIAMS VIVIENNE , PRINA MELLO ADRIELE , SHVETS IGOR , KELLEHER DERMOT , VOLKOV YURI
Published: January 2005

Abstract: Active T cell locomotion depends on efficient repeated cycles of integrin receptor/ligand interactions mediating cell adhesion and detachment, intracellular signaling cascades orchestrating posttranslation modifications of interacting proteins, dynamic reassembly of participating cytoskeletal elements, and structural support of associated scaffolding molecules. Using an integrated approach based on novel cutting edge technologies of live cell imaging, cell transfection, proteomics, and nanotechnology, we provide here a detailed characterisation of crucial mechanisms involved in LFA‐1 integrin‐mediated T cell migration. Polarization and phenotypic changes associated with LFA‐1‐triggered T cell locomotion is largely dependent on the intact functioning of the microtubule cytoskeleton. Experiments utilizing 4‐D (3‐D over time) confocal live imaging of T cells, microinjected with fully functional constructs encoding protein kinase C beta (PKC‐β) isoenzyme tagged with enhanced green fluorescent protein (GFP), elucidate that LFA‐1‐induced activation is associated with translocation of PKC‐β to sites associated with centrosomes and tubulin cytoskeleton in locomotory T lymphocytes. We also provide here a characterization of a novel microfluidics‐based multichannel platform enabling detailed analysis of leukocyte adhesion and migration under regulated shear stress conditions. Using precision machined surfaces, we demonstrate that the substrate topography can influence the motile response of the two different T cell types in different ways, and this can be quantified in terms of specified motility parameters. Finally, using an original in situ immunoprecipitation method, in which LFA‐1 antibodies are utilized to induce intracellular association of proteins in the cytoskeletal/signaling complex, we demonstrate that this complex includes a number of structural and signaling proteins, which have been identified by 2‐D electrophoresis and MALDI‐TOF protein sequencing.
Phase I Trial on sms‐D70 Somatostatin Analogue in Advanced Prostate and Renal Cell Cancer
Authors: JOENSUU T K , NILSSON S , HOLMBERG A R , MÁRQUEZ M , TENHUNEN M , SAARTO T , JOENSUU H
Published: January 2005

Abstract: Plasma concentrations and tolerability of a novel somatostatin analogue sms‐D70 were studied in patients with metastatic hormone‐resistant prostate cancer (HRPC) or metastatic renal cell cancer. To overcome the limitations of the octapeptides having affinity only to somatostatin receptor subtypes 2 and 5, HRPC expressing mainly somatostatin receptors 1 and 4, a somatostatin derivative based on the natural somatostatin having affinity to all five somatostatin receptor subtypes, was developed. The in vivo stability of this dextran‐conjugated derivative, somatostatin‐D70, was confirmed previously in animal studies, and the nanomolar “panaffinity” has been shown in in vitro receptor binding studies on cell lines transfected with the somatostatin receptor genes. Sms‐D70 was given with subcutaneous injection once a week at dose levels of 5, 10, 20, 35, and 50 mg. For pharmacokinetic studies, sms‐D70 was labeled with 131I. Fourteen patients were treated, of whom 10 had prostate and 4 renal cell cancer. The kinetic data revealed high stability with a long half‐life in the blood. The drug was well tolerated, and no grade 4 (WHO) toxicity was observed. The maximal tolerated dose could not be established due to the lack of dose‐limiting toxicities. Objective PSA responses were not recorded in these heavily treated patients, but subjective stabilization of pain was observed and urinary symptoms were alleviated in four patients. Three patients with metastatic HRPC received 5‐10‐mg intravenous injections of sms‐D70 once weekly for 4‐14 months on a compassionate use basis. In all cases, serum PSA values decreased more than 50% from the pretreatment level, but these results are difficult to interpret due to concomitant treatments given to these patients. In conclusion, sms‐D70 was well tolerated in the treatment of metastatic prostate and renal cell cancer, but no responses were found in these heavily treated patients.
Interaction between the Immune System and Tumor Cells: Cutaneous Disorders as a Consequence of Autoimmunity and Immunosuppression
Authors: LAIMER M , LANSCHUETZER C M , HINTNER H
Published: January 2005

Abstract: The interaction between the immune system and tumor cells is discussed with regard to examples of lymphoproliferative disorders associated with autoimmune phenomena, immunosuppression favoring tumorigenic infections, and a new therapeutic modality whereby a drug‐induced Th1 immune response leads to complete regression of skin tumors.
Secretory Leukocyte Protease Inhibitor in Cancer Development
Authors: DEVOOGDT NICK , REVETS HILDE , GHASSABEH GHOLAMREZA HASSANZADEH , DE BAETSELIER PATRICK
Published: January 2005

Abstract: Secretory leukocyte protease inhibitor (SLPI), an epithelial‐specific serine protease inhibitor of the whey acidic protein (WAP) family, exerts broad tissue‐protective functions: on the one hand, it counteracts inflammatory responses and invading pathogens, and on the other hand, it repairs the damaged tissue. Here, we report that subcutaneous in vivo passage of low‐malignant 3LL‐S cells increases the malignancy of these cancer cells. Applying the subtraction suppressive hybridization method to this cancer model, we identify SLPI as one of the genes whose level of expression directly correlates with the malignancy of the cancer cells. Using transfection experiments, we demonstrate that overexpression of mouse or human SLPI in 3LL‐S cells is sufficient to enhance their malignant behavior, and that this activity of SLPI is related to its ability to inhibit serine proteases. Furthermore, using cDNA dot‐blot hybridization, we show that in human gynecological cancer tissue SLPI expression is frequently increased as compared with normal tissue. This study underscores the promalignant role of SLPI in the development of cancer and its potential usefulness as a biomarker for gynecological cancer.
Targeting Multiple Myeloma Cells and Their Bone Marrow Microenvironment
Authors: PAGNUCCO GUIDO , CARDINALE GIOVANNI , GERVASI FRANCESCO
Published: January 2005

Abstract: Although multiple myeloma (MM) is sensitive to chemotherapy and radiation therapy, long‐term disease‐free survival is rare, and MM remains incurable despite conventional and high‐dose therapies. Direct (cell‐cell contact) and soluble (via cytokines) forms of interactions between MM cells and bone marrow stroma regulate growth, survival, and homing of MM cells. These interactions also play a critical role in angiogenesis and in myeloma bone disease. In recent years, several studies have established the biologic significance of cytokines in MM pathogenesis and delineated signaling cascades mediating their effects, providing the framework for related novel therapies targeting not only the MM cell, but also the bone marrow microenvironment.