bluuebunny/crossref_metadata_2025 huggingface dataset:
Developmental expression of the alpha-skeletal actin gene
Authors: Bertola Laura D , Ott Elisabeth B , Griepsma Sander , Vonk Freek J , Bagowski Christoph P
Published: June 2008

Background

Actin is a cytoskeletal protein which exerts a broad range of functions in almost all eukaryotic cells. In higher vertebrates, six primary actin isoforms can be distinguished: alpha-skeletal, alpha-cardiac, alpha-smooth muscle, gamma-smooth muscle, beta-cytoplasmic and gamma-cytoplasmic isoactin. Expression of these actin isoforms during vertebrate development is highly regulated in a temporal and tissue-specific manner, but the mechanisms and the specific differences are currently not well understood. All members of the actin multigene family are highly conserved, suggesting that there is a high selective pressure on these proteins.

Results

We present here a model for the evolution of the genomic organization of alpha-skeletal actin and by molecular modeling, illustrate the structural differences of actin proteins of different phyla. We further describe and compare alpha-skeletal actin expression in two developmental stages of five vertebrate species (mouse, chicken, snake, salamander and fish). Our findings confirm that alpha-skeletal actin is expressed in skeletal muscle and in the heart of all five species. In addition, we identify many novel non-muscular expression domains including several in the central nervous system.

Conclusion

Our results show that the high sequence homology of alpha-skeletal actins is reflected by similarities of their 3 dimensional protein structures, as well as by conserved gene expression patterns during vertebrate development. Nonetheless, we find here important differences in 3D structures, in gene architectures and identify novel expression domains for this structural and functional important gene.
Developing and applying a gene functional association network for anti-angiogenic kinase inhibitor activity assessment in an angiogenesis co-culture model
Authors: Chen Yuefeng , Wei Tao , Yan Lei , Lawrence Frank , Qian Hui-Rong , Burkholder Timothy P , Starling James J , Yingling Jonathan M , Shou Jianyong
Published: June 2008

Background

Tumor angiogenesis is a highly regulated process involving intercellular communication as well as the interactions of multiple downstream signal transduction pathways. Disrupting one or even a few angiogenesis pathways is often insufficient to achieve sustained therapeutic benefits due to the complexity of angiogenesis. Targeting multiple angiogenic pathways has been increasingly recognized as a viable strategy. However, translation of the polypharmacology of a given compound to its antiangiogenic efficacy remains a major technical challenge. Developing a global functional association network among angiogenesis-related genes is much needed to facilitate holistic understanding of angiogenesis and to aid the development of more effective anti-angiogenesis therapeutics.

Results

We constructed a comprehensive gene functional association network or interactome by transcript profiling an in vitro angiogenesis model, in which human umbilical vein endothelial cells (HUVECs) formed capillary structures when co-cultured with normal human dermal fibroblasts (NHDFs). HUVEC competence and NHDF supportiveness of cord formation were found to be highly cell-passage dependent. An enrichment test of Biological Processes (BP) of differentially expressed genes (DEG) revealed that angiogenesis related BP categories significantly changed with cell passages. Built upon 2012 DEGs identified from two microarray studies, the resulting interactome captured 17226 functional gene associations and displayed characteristics of a scale-free network. The interactome includes the involvement of oncogenes and tumor suppressor genes in angiogenesis. We developed a network walking algorithm to extract connectivity information from the interactome and applied it to simulate the level of network perturbation by three multi-targeted anti-angiogenic kinase inhibitors. Simulated network perturbation correlated with observed anti-angiogenesis activity in a cord formation bioassay.

Conclusion

We established a comprehensive gene functional association network to model in vitro angiogenesis regulation. The present study provided a proof-of-concept pilot of applying network perturbation analysis to drug phenotypic activity assessment.
Genome-wide survey of allele-specific splicing in humans
Authors: Nembaware Victoria , Lupindo Bukiwe , Schouest Katherine , Spillane Charles , Scheffler Konrad , Seoighe Cathal
Published: June 2008

Background

Accurate mRNA splicing depends on multiple regulatory signals encoded in the transcribed RNA sequence. Many examples of mutations within human splice regulatory regions that alter splicing qualitatively or quantitatively have been reported and allelic differences in mRNA splicing are likely to be a common and important source of phenotypic diversity at the molecular level, in addition to their contribution to genetic disease susceptibility. However, because the effect of a mutation on the efficiency of mRNA splicing is often difficult to predict, many mutations that cause disease through an effect on splicing are likely to remain undiscovered.

Results

We have combined a genome-wide scan for sequence polymorphisms likely to affect mRNA splicing with analysis of publicly available Expressed Sequence Tag (EST) and exon array data. The genome-wide scan uses published tools and identified 30,977 SNPs located within donor and acceptor splice sites, branch points and exonic splicing enhancer elements. For 1,185 candidate splicing polymorphisms the difference in splicing between alternative alleles was corroborated by publicly available exon array data from 166 lymphoblastoid cell lines. We developed a novel probabilistic method to infer allele-specific splicing from EST data. The method uses SNPs and alternative mRNA isoforms mapped to EST sequences and models both regulated alternative splicing as well as allele-specific splicing. We have also estimated heritability of splicing and report that a greater proportion of genes show evidence of splicing heritability than show heritability of overall gene expression level. Our results provide an extensive resource that can be used to assess the possible effect on splicing of human polymorphisms in putative splice-regulatory sites.

Conclusion

We report a set of genes showing evidence of allele-specific splicing from an integrated analysis of genomic polymorphisms, EST data and exon array data, including several examples for which there is experimental evidence of polymorphisms affecting splicing in the literature. We also present a set of novel allele-specific splicing candidates and discuss the strengths and weaknesses of alternative technologies for inferring the effect of sequence variants on mRNA splicing.
Vav3 oncogene activates estrogen receptor and its overexpression may be involved in human breast cancer
Authors: Lee Kiwon , Liu Yin , Mo Jun Qin , Zhang Jinsong , Dong Zhongyun , Lu Shan
Published: June 2008

Background

Our previous study revealed that Vav3 oncogene is overexpressed in human prostate cancer, activates androgen receptor, and stimulates growth in prostate cancer cells. The current study is to determine a potential role of Vav3 oncogene in human breast cancer and impact on estrogen receptor a (ERα)-mediated signaling axis.

Methods

Immunohistochemistry analysis was performed in 43 breast cancer specimens and western blot analysis was used for human breast cancer cell lines to determine the expression level of Vav3 protein. The impact of Vav3 on breast cancer cell growth was determined by siRNA knockdown of Vav3 expression. The role of Vav3 in ERα activation was examined in luciferase reporter assays. Deletion mutation analysis of Vav3 protein was performed to localize the functional domain involved in ERα activation. Finally, the interaction of Vav3 and ERα was assessed by GST pull-down analysis.

Results

We found that Vav3 was overexpressed in 81% of human breast cancer specimens, particularly in poorly differentiated lesions. Vav3 activated ERα partially via PI3K-Akt signaling and stimulated growth of breast cancer cells. Vav3 also potentiated EGF activity for cell growth and ERα activation in breast cancer cells. More interestingly, we found that Vav3 complexed with ERα. Consistent with its function for AR, the DH domain of Vav3 was essential for ERα activation.

Conclusion

Vav3 oncogene is overexpressed in human breast cancer. Vav3 complexes with ERα and enhances ERα activity. These findings suggest that Vav3 overexpression may aberrantly enhance ERα-mediated signaling axis and play a role in breast cancer development and/or progression.
Perioperative celecoxib administration for pain management after total knee arthroplasty – A randomized, controlled study
Authors: Huang Yu-Min , Wang Chiu-Meng , Wang Chen-Ti , Lin Wei-Peng , Horng Lih-Ching , Jiang Ching-Chuan
Published: June 2008

Background

Non-steroidal anti-inflammatory drugs (NSAIDs) are recommended for multimodal postoperative pain management. We evaluated opioid-sparing effects and rehabilitative results after perioperative celecoxib administration for total knee arthroplasty.

Methods

This was a prospective, randomized, observer-blind control study. Eighty patients that underwent total knee arthroplasty were randomized into two groups of 40 each. The study group received a single 400 mg dose of celecoxib, one hour before surgery, and 200 mg of celecoxib every 12 hours for five days, along with patient-controlled analgesic (PCA) morphine. The control group received only PCA morphine for postoperative pain management. Visual analog scale (VAS) pain scores, active range of motion (ROM), total opioid use and postoperative nausea/vomiting were analyzed.

Results

Groups were comparable for age, pre-operative ROM, operation duration and intraoperative blood loss. Resting VAS pain scores improved significantly in the celecoxib group, compared with controls, at 48 hrs (2.13 ± 1.68 vs. 3.43 ± 1.50, p = 0.03) and 72 hrs (1.78 ± 1.66 vs. 3.17 ± 2.01, p = 0.02) after surgery. Active ROM also increased significantly in the patients that received celecoxib, especially in the first 72 hrs [40.8° ± 17.3° vs. 25.8° ± 11.5°, p = 0.01 (day 1); 60.7° ± 18.1° vs. 45.0° ± 17.3°, p = 0.004 (day 2); 77.7° ± 15.1° vs. 64.3° ± 16.9°, p = 0.004 (day 3)]. Opioid requirements decreased about 40% (p = 0.03) in the celecoxib group. Although patients suffering from post-operative nausea/vomiting decreased from 43% in control group to 28% in celecoxib group, this was not significant (p = 0.57). There were no differences in blood loss (intra- and postoperative) between the groups. Celecoxib resulted in no significant increase in the need for blood transfusions.

Conclusion

Perioperative celecoxib significantly improved postoperative resting pain scores at 48 and 72 hrs, opioid consumption, and active ROM in the first three days after total knee arthroplasty, without increasing the risks of bleeding.

Trial registration

Clinicaltrials.gov NCT00598234
Prevalence of putative virulence factors and antimicrobial susceptibility of Enterococcus faecalis isolates from patients with dental Diseases
Authors: Salah Randa , Dar-Odeh Najla , Abu Hammad Osama , Shehabi Asem A
Published: June 2008

Background

This study investigated the prevalence of Enterococcus faecalis, its putative virulence factors and antimicrobial susceptibility in individuals with and without dental diseases. A total of 159 oral rinse specimens were collected from patients (n = 109) suffering from dental diseases and healthy controls (n = 50).

Results

E. faecalis was detected using only culture in 8/109 (7.3%) of the patients with various types of dental diseases, whereas no E. faecalis was found in the healthy controls weather using both culture and PCR. Phenotype characterizations of the 8 E. faecalis isolates indicated that 25% of the isolates produced haemolysin and 37.5% produced gelatinase. Most important virulence genes; collagen binding protein (ace) and endocarditis antigen (efaA) were present in all 8 E. faecalis isolates, while haemolysin activator gene (cylA) was detected only in 25% of isolates, and all isolates were negative for esp gene. All E. faecalis isolates were 100% susceptible to ampicillin, chloramphenicol, ciprofloxacin, vancomycin, and teicoplanin, and to less extent to erythromycin (62.5%).

Conclusion

This study shows that all E. faecalis isolates were recovered only from patients with dental diseases especially necrotic pulps, and all isolates carried both collagen binding protein and endocarditis antigen genes and highly susceptible to frequently used antimicrobial drugs in Jordan.
How do parents of children with juvenile idiopathic arthritis (JIA) perceive their therapies?
Authors: Rouster-Stevens Kelly , Nageswaran Savithri , Arcury Thomas A , Kemper Kathi J
Published: June 2008

Background

Complementary and alternative medical (CAM) therapies are commonly used by pediatric patients with chronic medical conditions. Little is known about parents' perceptions of these therapies. This study describes the views of parents of patients with juvenile idiopathic arthritis (JIA) regarding conventional and CAM therapies.

Methods

Parents of children with JIA seen at a pediatric rheumatology clinic were surveyed between June 1 and July 31, 2007. Questionnaires asked about patients' use of over 75 therapies in the past 30 days, their perceived helpfulness (0 = not helpful; 3 = very helpful), perceived side effects (0 = none; 3 = severe), and whether each therapy would be recommended to other patients with JIA (Yes, No, Not sure).

Results

Questionnaires were returned by 52/76 (68%) parents; patients' average age was 10.9 years and 87% were Caucasian. Medications were used by 45 (88%) patients; heat (67%) and extra rest (54%) were also commonly used. CAM therapies were used by 48 (92%), e.g., massage (54%), vitamins and other supplements (54%), avoiding foods that worsened pain (35%) and stress management techniques (33%). Among the therapies rated by 3 or more parents, those that scored 2.5 or higher on helpfulness were: biologic medications, methotrexate, naproxen, wheelchairs, orthotics, heat, vitamins C and D, music, support groups and prayer. CAM therapies had 0 median side effects and parents would recommend many of them to other families.

Conclusion

JIA patients use diverse therapies. Parents report that many CAM therapies are helpful and would recommend them to other parents. These data can be used in counseling patients and guiding future research.
Water disinfection by-products and the risk of specific birth defects: a population-based cross-sectional study in Taiwan
Authors: Hwang Bing-Fang , Jaakkola Jouni JK , Guo How-Ran
Published: June 2008

Background

Recent findings suggest that exposure to disinfection by-products may increase the risk of birth defects. Previous studies have focused mainly on birth defects in general or groups of defects. The objective of the present study was to assess the effect of water disinfection by-products on the risk of most common specific birth defects.

Methods

We conducted a population-based cross-sectional study of 396,049 Taiwanese births in 2001–2003 using information from the Birth Registry and Waterworks Registry. We compared the risk of eleven most common specific defects in four disinfection by-product exposure categories based on the levels of total trihalomethanes (TTHMs) representing high (TTHMs 20+ μg/L), medium (TTHMs 10–19 μg/L), low exposure (TTHMs 5–9 μg/L), and 0–4 μg/L as the reference category. In addition, we conducted a meta-analysis of the results from the present and previous studies focusing on the same birth defects.

Results

In multivariate logistic regression analysis the risk of ventricular septal defects (adjusted odds ratio 1.81, 95% confidence interval: 0.98 3.35), cleft palate (1.56. 95% CI: 1.00, 2.41), and anencephalus (1.96, 95% CI: 0.94, 4.07) were elevated in the high exposure compared to the reference category. In the meta-analysis, the summary odds ratio for ventricular septal defects (1.59, 95% CI: 1.21, 2.07) was consistently elevated.

Conclusion

The present study suggests that prenatal exposure to disinfection by-products increases the risk of ventricular septal defects, cleft palate, and anencephalus. The evidence on ventricular septal defects is consistent in the three available studies.
Re-evaluation of blood mercury, lead and cadmium concentrations in the Inuit population of Nunavik (Québec): a cross-sectional study
Authors: Fontaine Julie , Dewailly Éric , Benedetti Jean-Louis , Pereg Daria , Ayotte Pierre , Déry Serge
Published: June 2008

Background

Arctic populations are exposed to mercury, lead and cadmium through their traditional diet. Studies have however shown that cadmium exposure is most often attributable to tobacco smoking. The aim of this study is to examine the trends in mercury, lead and cadmium exposure between 1992 and 2004 in the Inuit population of Nunavik (Northern Québec, Canada) using the data obtained from two broad scale health surveys, and to identify sources of exposure in 2004.

Methods

In 2004, 917 adults aged between 18 and 74 were recruited in the 14 communities of Nunavik to participate to a broad scale health survey. Blood samples were collected and analysed for metals by inductively coupled plasma mass spectrometry, and dietary and life-style characteristics were documented by questionnaires. Results were compared with data obtained in 1992, where 492 people were recruited for a similar survey in the same population.

Results

Mean blood concentration of mercury was 51.2 nmol/L, which represent a 32% decrease (p < 0.001) between 1992 and 2004. Mercury blood concentrations were mainly explained by age (partial r2 = 0.20; p < 0.0001), and the most important source of exposure to mercury was marine mammal meat consumption (partial r2 = 0.04; p < 0.0001). In 2004, mean blood concentration of lead was 0.19 μmol/L and showed a 55% decrease since 1992. No strong associations were observed with any dietary source, and lead concentrations were mainly explained by age (partial r2 = 0.20.; p < 0.001). Blood cadmium concentrations showed a 22% decrease (p < 0.001) between 1992 and 2004. Once stratified according to tobacco use, means varied between 5.3 nmol/L in never-smokers and 40.4 nmol/L in smokers. Blood cadmium concentrations were mainly associated with tobacco smoking (partial r2 = 0.56; p < 0.0001), while consumption of caribou liver and kidney remain a minor source of cadmium exposure among never-smokers.

Conclusion

Important decreases in mercury, lead and cadmium exposure were observed. Mercury decrease could be explained by dietary changes and the ban of lead cartridges use likely contributed to the decrease in lead exposure. Blood cadmium concentrations remain high and, underscoring the need for intensive tobacco smoking prevention campaigns in the Nunavik population.
Comparison of the self-administered and interviewer-administered modes of the child-OIDP
Authors: Tsakos Georgios , Bernabé Eduardo , O'Brien Kevin , Sheiham Aubrey , de Oliveira Cesar
Published: June 2008

Background

The mode of questionnaire administration may affect the estimates and applicability of oral health-related quality of life indicators. The aim of this study was to compare psychometrically the self-administered Child-OIDP index with the original interviewer-administered instrument.

Methods

This was a cross-sectional study of 144 consecutive children aged 9–16 years referred to orthodontic clinics in Bedfordshire. To compare the two administration modes of the Child-OIDP, the sample was randomly split in two groups. The two groups were analysed in terms of baseline characteristics, self-perceived measures (self-rated oral health, self-perceived need for braces, happiness with dental appearance, frequency of thinking about dental appearance), Child-OIDP performance scores and overall score and psychometric properties (criterion validity and internal reliability).

Results

No significant difference between the two groups was found in relation to their sociodemographic profile and self-perceived measures. The self- and interviewer-administered Child-OIDP had identical mean scores and did not differ in recording any of the eight performances (p ≥ 0.206). For criterion validity, the correlation coefficients of the Child-OIDP with self-perceived measures were not different between the two modes of administration (p ≥ 0.118). Furthermore, the Cronbach's alpha values of the two groups were similar (p = 0.466).

Conclusion

This study demonstrated that the self-administered Child-OIDP performed the same as the original interviewer-administered mode, while at the same time reducing administration burden. This provides support for the use of the self-administered Child-OIDP. Further studies should focus on a more comprehensive psychometric evaluation.
GRK5 deficiency exaggerates inflammatory changes in TgAPPsw mice
Authors: Li Longxuan , Liu Jun , Suo William Z
Published: June 2008

Background

Deficiency of membrane G-protein coupled receptor (GPCR) kinase-5 (GRK5) recently has been linked to early AD pathogenesis, and has been suggested to contribute to augmented microglial activation in vitro by sensitizing relevant GPCRs. However, GRK5 deficient mice did not show any signs of microgliosis, except for their moderate increase in axonal defects and synaptic degenerative changes during aging. We have speculated that one possible reason for the absence of microgliosis in these animals might be due to lack of an active inflammatory process involving activated GPCR signaling, since GRKs only act on activated GPCRs. The objective of this study was to determine whether the microgliosis is exaggerated in TgAPPsw (Tg2576) mice also deficient in GRK5, in which fibrillar β-amyloid (Aβ) and an active inflammatory process involving activated GPCR signaling are present.

Methods

Both quantitative and qualitative immunochemistry methods were used to evaluate the microgliosis and astrogliosis in these animals.

Results

We found that inactivation of one copy of the GRK5 gene in the TgAPPsw mice resulted in approximately doubled extent of microgliosis, along with significantly exaggerated astrogliosis, in both hippocampus and cortex of the aged animals. Consistent with previous observations, the activated microglia were located primarily near or surrounding the fibrillar Aβ deposits.

Conclusion

The results demonstrate that GRK5 deficiency in vivo significantly exaggerates microgliosis and astrogliosis in the presence of an inflammatory initiator, such as the excess fibrillar Aβ and the subsequent active inflammatory reactions in the TgAPPsw mice.
Physico-chemical modifications of conjugated linoleic acid for ruminal protection and oxidative stability
Authors: Moon Hyun-Seuk , Lee Hong-Gu , Chung Chung-Soo , Choi Yun-Jaie , Cho Chong-Su
Published: June 2008

Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of octadecadienoic acid [linoleic acid (LA), 18:2n-6]. Although ruminant milk and meat products represent the largest natural source of CLA and therefore, their concentration in ruminant lipids are of interest to human health, chemical or physical modifications of CLA should be needed as a means to enhance oxidative stability, to improve post-ruminal bioavailability, and to increase the clinical application. In fact, CLA are rapidly decomposed to form furan fatty acids when its are oxidized in air, and the effectiveness of dietary supplements of CLA may be related to the extent that their metabolisms by rumen bacteria are avoided. For these reasons, many scientists have examined the effect of manufacturing and protection on the stability of CLA in ruminants and food products. In this review, physico-chemical modifications of CLA for ruminal protection such as calcium salt (Ca), formaldehyde protection (FP), lipid encapsulation (LE), and amide linkage (AL), and for oxidative stability such as green tea catechin (GTC), cyclodextrin (CD), arginine (Arg), amylase, and PEGylation are proposed.
Compound Heterozygosity in Sodium Channel Nav1.7 in a Family with Hereditary Erythermalgia
Authors: Samuels Mark E , Morsche Rene HM te , Lynch Mary E , Drenth Joost PH
Published: June 2008

Hereditary erythermalgia is a painful and debilitating genetic disorder associated with mutations in voltage-gated sodium channel Nav1.7. We have previously reported a Canadian family segregating erythermalgia consistently with a dominant genetic etiology. Molecular analysis of the proband from the family detected two different missense mutations in Nav1.7. In the present study we have performed a long-term follow-up clinical study of disease progression in three affected family members. A more extensive molecular study has also been completed, analyzing the segregation of the two missense variants in the family. The two variants (P610T, L858F) segregate independently with respect to clinical presentation. Detailed genotype/phenotype correlation suggests that one of the two variants (L858F) is causal for erythermalgia. The second variant (P610T) may modify the phenotype in the proband. This is the second reported study of potential compound heterozygosity for coding polymorphisms in Nav1.7, the first being in a patient with paroxysmal extreme pain disorder.
Reconstruction of the gastric passage by a side-to-side gastrogastrostomy after failed vertical-banded gastroplasty: a case report
Authors: Soll Christopher , Müller Markus K. , Wildi Stefan , Clavien Pierre-Alain , Weber Markus
Published: June 2008

Introduction

Vertical-banded gastroplasty, a technique that is commonly performed in the treatment of morbid obesity, represents a nonadjustable restrictive procedure which reduces the volume of the upper stomach by a vertical stapler line. In addition, a textile or silicone band restricts food passage through the stomach.

Case presentation

A 71-year-old woman presented with a severe gastric stenosis 11 years after vertical gastroplasty. We describe a side-to-side gastrogastrostomy as a safe surgical procedure to restore the physiological gastric passage after failed vertical-banded gastroplasty.

Conclusion

Occasionally, restrictive procedures for morbid obesity cannot be converted into an alternative bariatric procedure to maintain weight control. This report demonstrates that a side-to-side gastrogastrostomy is a feasible and safe procedure.
Association of ABCB1genetic variants with renal function in Africans and in Caucasians
Authors: Bochud Murielle , Eap Chin B , Maillard Marc , Johnson Toby , Vollenweider Peter , Bovet Pascal , Elston Robert C , Bergmann Sven , Beckmann Jacques S , Waterworth Dawn M , Mooser Vincent , Gabriel Anne , Burnier Michel
Published: June 2008

Background

The P-glycoprotein, encoded by theABCB1gene, is expressed in human endothelial and mesangial cells, which contribute to control renal plasma flow and glomerular filtration rate. We investigated the association ofABCB1variants with renal function in African and Caucasian subjects.

Methods

In Africans (290 subjects from 62 pedigrees), we genotyped the2677G>Tand3435 C>T ABCB1polymorphisms. Glomerular filtration rate (GFR) was measured using inulin clearance and effective renal plasma flow (ERPF) using para-aminohippurate clearance. In Caucasians (5382 unrelated subjects), we analyzed 30 SNPs located within and aroundABCB1, using data from the Affymetrix 500 K chip. GFR was estimated using the simplified Modification of the Diet in Renal Disease (MDRD) and Cockcroft-Gault equations.

Results

In Africans, compared to the reference genotype (GG or CC), each copy of the2677Tand3435Tallele was associated, respectively, with: GFR higher by 10.6 ± 2.9 (P< 0.001) and 4.4 ± 2.3 (P= 0.06) mL/min; ERPF higher by 47.5 ± 11.6 (P< 0.001) and 28.1 ± 10.5 (P= 0.007) mL/min; and renal resistances lower by 0.016 ± 0.004 (P< 0.001) and 0.011 ± 0.004 (P= 0.004) mm Hg/mL/min. In Caucasians, we identified 3 polymorphisms in theABCB1gene that were strongly associated with all estimates of GFR (smallest P value = 0.0006, overall P = 0.014 after multiple testing correction).

Conclusion

Variants of theABCB1gene were associated with renal function in both Africans and Caucasians and may therefore confer susceptibility to nephropathy in humans. If confirmed in other studies, these results point toward a new candidate gene for nephropathy in humans.
Identification of novel monosodium urate crystal regulated mRNAs by transcript profiling of dissected murine air pouch membranes
Authors: Pessler Frank , Mayer Christian T , Jung Sung Mun , Behrens Ed M , Dai Lie , Menetski Joseph P , Schumacher H Ralph
Published: June 2008

Introduction

The murine air pouch is a bursa-like space that resembles the human synovial membrane. Injection of monosodium urate (MSU) crystals into the pouch elicits an acute inflammatory response similar to human gout. We conducted the present study to identify mRNAs that were highly regulated by MSU crystals in the pouch membrane.

Methods

Air pouch membranes were meticulously dissected away from the overlying skin. Gene expression differences between MSU crystal stimulated and control membranes were determined by oligonucleotide microarray analysis 9 hours after injection of MSU crystals or buffer only. Differential regulation of selected targets was validated by relative quantitative PCR in time course experiments with dissected air pouch membranes and murine peritoneal macrophages.

Results

Eleven of the 12 most highly upregulated mRNAs were related to innate immunity and inflammation. They included mRNAs encoding histidine decarboxylase (the enzyme that synthesizes histamine), IL-6, the cell surface receptors PUMA-g and TREM-1, and the polypeptides Irg1 and PROK-2. IL-6 mRNA rose 108-fold 1 hour after crystal injection, coinciding with a surge in mRNAs encoding IL-1β, tumour necrosis factor-α and the immediate early transcription factor Egr-1. The other mRNAs rose up to 200-fold within the subsequent 3 to 8 hours. MSU crystals induced these mRNAs in a dose-dependent manner in cultured macrophages, with similar kinetics but lower fold changes. Among the downregulated mRNAs, quantitative PCR confirmed significant decreases in mRNAs encoding TREM-2 (an inhibitor of macrophage activation) and granzyme D (a constituent of natural killer and cytotoxic T cells) within 50 hours after crystal injection.

Conclusion

This analysis identified several genes that were previously not implicated in MSU crystal inflammation. The marked rise of the upregulated mRNAs after the early surge in cytokine and Egr-1 mRNAs suggests that they may be part of a 'second wave' of factors that amplify or perpetuate inflammation. Transcript profiling of the isolated air pouch membrane promises to be a powerful tool for identifying genes that act at different stages of inflammation.
Mammary carcinoma behavior is programmed in the precancer stem cell
Authors: Damonte Patrizia , Hodgson J Graeme , Chen Jane Qian , Young Lawrence JT , Cardiff Robert D , Borowsky Alexander D
Published: June 2008

Introduction

The 'MINO' (mammary intraepithelial neoplasia outgrowth) mouse model of ductal carcinoma in situ (DCIS) consists of six lines with distinct morphologic phenotypes and behavior, each meeting experimentally defined criteria for 'precancer'. Specifically, these lines grow orthotopically in cleared mammary fat pads and consistently progress to an invasive phenotype that is capable of ectopic growth. Transition to carcinoma has a consistent latency for each line, and three of the lines also exhibit pulmonary metastatic potential.

Methods

Gland cleared orthotopic transplanted precancer MINO tissues were analyzed by bacterial artifical chromosome and oligo array comparative genomic hybridization, microsatellite PCR, and telomerase repeat amplification assay. MINO cells were dissociated and cultured in three dimensional culture and transplanted in syngeneic gland cleared mammary fat pads.

Results

Comparative genomic hybridization shows that the precancer and invasive tumors are genetically stable, with low level changes including whole chromosome gains in some lines. No changes are associated with progression, although spontaneous focal amplifications and deletions were detected occasionally. Microsatellite analysis shows a low frequency of alterations that are predominantly permanent within a MINO line. Telomerase activity is increased in both the MINO and the derived tumors when compared with normal mouse mammary gland. Dissociation of the precancer lesion cells and three dimensional 'spheroid' culture of single cells reveals a bipotential for myoepithelial and luminal differentiation and the formation of unique three-dimensional 'MINOspheres'. These MINOspheres exhibit features that are intermediate between spheroids that are derived from normal and carcinoma cells. Transplantation of a single cell derived MINOsphere recapitulates the outgrowth of the precancer morphology and progression to carcinoma.

Conclusion

These data establish a precancer 'stem' cell that is capable of self-renewal and multilineage differentiation as the origin of invasive cancer. Within the context of this model, these cells have programmed potential for latency and metastasis that does not appear to require sequential genetic 'hits' for transformation.
Optic nerve sonography in the diagnostic evaluation of adult brain injury
Authors: Soldatos Theodoros , Karakitsos Dimitrios , Chatzimichail Katerina , Papathanasiou Matilda , Gouliamos Athanasios , Karabinis Andreas
Published: June 2008

Introduction

The optic nerve sheath diameter (ONSD) may be increased in brain-injured patients, especially children, with intracranial hypertension. We investigated whether measurements of ONSD correlated with simultaneous noninvasive and invasive measurements of the intracranial pressure (ICP) in brain-injured adults.

Methods

Seventy-six critical care patients (58 males; 47 ± 18 years old) were included in the study. Fifty patients suffered from brain injury, whereas 26 had no intracranial pathology and served as control individuals. Initially, brain-injured patients were evaluated clinically (Glasgow Coma Scale) and using a semiquantitative (I to VI) neuroimaging scale (Marshall Scale). Thereafter, the patients were divided into those with moderate (Marshall Scale = I and Glasgow Coma Scale > 8 [n = 18]) and severe (Marshall Scale = II to VI and Glasgow Coma Scale ≤8 [n = 32]) brain injury. All patients underwent noninvasive measurement of the ICP (estimated ICP) by transcranial Doppler sonography, and synchronous ONSD measurements by optic nerve sonography. Finally, invasive ICP measurement using an intraparenchymal catheter was performed in patients with severe brain injury.

Results

ONSD and estimated ICP were both significantly increased (6.1 ± 0.7 mm and 26.2 ± 8.7 mmHg, respectively; P < 0.0001) in patients with severe brain injury as compared with patients with moderate brain injury (4.2 ± 1.2 mm and 12.0 ± 3.6 mmHg) and compared with control individuals (3.6 ± 0.6 mm and 10.3 ± 3.1 mmHg). Furthermore, in patients with severe brain injury the ONSD measurements were strongly correlated with estimated ICP values (r = 0.80, P < 0.0001) as well as with the neuroimaging scale results (r = 0.82, P < 0.001). In the patients with severe brain injury, ONSD measurements correlated with invasive ICP values (r = 0.68, P = 0.002). The best cut-off value of ONSD for predicting elevated ICP was 5.7 mm (sensitivity = 74.1% and specificity = 100%).

Conclusion

ONSD measurements correlate with noninvasive and invasive measurements of the ICP, and with head computed tomography scan findings in brain-injured adults. Hence, optic nerve sonography may serve as an additional diagnostic tool that could alert clinicians to the presence of elevated ICP, whenever invasive ICP evaluation is contraindicated and/or is not available. This trial is International Standard Randomised Controlled Trial Number registered (ISRCTN 91941687).
Gene expression profiling of human prostate cancer stem cells reveals a pro-inflammatory phenotype and the importance of extracellular matrix interactions
Authors: Birnie Richard , Bryce Steven D , Roome Claire , Dussupt Vincent , Droop Alastair , Lang Shona H , Berry Paul A , Hyde Catherine F , Lewis John L , Stower Michael J , Maitland Norman J , Collins Anne T
Published: June 2008

Background

The tumor-initiating capacity of many cancers is considered to reside in a small subpopulation of cells (cancer stem cells). We have previously shown that rare prostate epithelial cells with a CD133+/α2β1hi phenotype have the properties of prostate cancer stem cells. We have compared gene expression in these cells relative to their normal and differentiated (CD133-/α2β1low) counterparts, resulting in an informative cancer stem cell gene-expression signature.

Results

Cell cultures were generated from specimens of human prostate cancers (n = 12) and non-malignant control tissues (n = 7). Affymetrix gene-expression arrays were used to analyze total cell RNA from sorted cell populations, and expression changes were selectively validated by quantitative RT-PCR, flow cytometry and immunocytochemistry. Differential expression of multiple genes associated with inflammation, cellular adhesion, and metastasis was observed. Functional studies, using an inhibitor of nuclear factor κB (NF-κB), revealed preferential targeting of the cancer stem cell and progenitor population for apoptosis whilst sparing normal stem cells. NF-κB is a major factor controlling the ability of tumor cells to resist apoptosis and provides an attractive target for new chemopreventative and chemotherapeutic approaches.

Conclusion

We describe an expression signature of 581 genes whose levels are significantly different in prostate cancer stem cells. Functional annotation of this signature identified the JAK-STAT pathway and focal adhesion signaling as key processes in the biology of cancer stem cells.
CSF-1 receptor structure/function in MacCsf1r–/– macrophages: regulation of proliferation, differentiation, and morphology
Authors: Yu Wenfeng , Chen Jian , Xiong Ying , Pixley Fiona J , Dai Xu-Ming , Yeung Yee-Guide , Stanley E Richard
Published: June 2008

CSF-1 is the major regulator of tissue macrophage development and function. A GM-CSF-dependent, CSF-1 receptor (CSF-1R)-deficient F4/80hiMac-1+Gr1–CD11c+ bone marrow macrophage (BMM) line (MacCsf1r−/−) was developed to study the roles of the eight intracellular CSF-1R tyrosines phosphorylated upon receptor activation. Retroviral expression of the wild-type CSF-1R rescued the CSF-1-induced survival, proliferation, differentiation, and morphological characteristics of primary BMM. Mutation of all eight tyrosines failed to rescue, whereas the individual Y → F mutants (544, 559, 697, 706, 721, 807, 921, 974) rescued these CSF-1-inducible phenotypes to varying degrees. The juxtamembrane domain Y559F and activation loop Y807F mutations severely compromised proliferation and differentiation, whereas Y706, Y721F, and Y974F mutations altered morphological responses, and Y706F increased differentiation. Despite their retention of significant in vitro tyrosine kinase activity, Y559F and Y807F mutants exhibited severely impaired in vivo receptor tyrosine phosphorylation, consistent with the existence of cellular mechanisms inhibiting CSF-1R tyrosine phosphorylation that are relieved by phosphorylation of these two sites. The MacCsf1r−/− macrophage line will facilitate genetic and proteomic approaches to CSF-1R structure/function studies in the major disease-related CSF-1R-expressing cell type.
Regulation of superoxide production in neutrophils: role of calcium influx
Authors: Bréchard Sabrina , Tschirhart Eric J
Published: June 2008

Upon stimulation, activation of NADPH oxidase complexes in neutrophils produces a burst of superoxide anions contributing to oxidative stress and the development of inflammatory process. Store-operated calcium entry (SOCE), whereby the depletion of intracellular stores induces extracellular calcium influx, is known to be a crucial element of NADPH oxidase regulation. However, the mechanistic basis mediating SOCE is still only partially understood, as is the signal-coupling pathway leading to modulation of store-operated channels. This review emphasizes the role of calcium influx in the control of the NADPH oxidase and summarizes the current knowledge of pathways mediating this extracellular calcium entry in neutrophils. Such investigations into the cross-talk between NADPH oxidase and calcium might allow the identification of novel pharmacological targets with clinical use, particularly in inflammatory diseases.
Lymphatic Tissue Engineering
Authors: Hitchcock Thomas , Niklason Laura
Published: June 2008

In the last 5 years major advances have been made in the field of tissue engineering. However, while engineering of tissues from nearly every major system in the body have been studied and improved, little has been done with the engineering of viable lymphatic tissues. Recent advances in understanding of lymphatic biology have allowed the easy isolation of pure lymphatic cell cultures, increasing, in turn, the ability to study lymphatic biology in greater detail. This has allowed the elucidation of lymphatic properties on the structural, cellular, and molecular levels, making possible the successful development of the first lymphatic engineered tissues. Among such advances are the engineering of lymphatic capillaries, the development of a functioning bioreactor designed to culture lymph nodes in vitro, and in vivo growth of lymphatic organoids. However, there has been no research on the engineering of functional lymphangions. While the advances made in the study of lymphatic biology are encouraging, the complexities of the system make the engineering of certain functional lymphatic tissues somewhat more difficult.
Animal Models for the Molecular and Mechanistic Study of Lymphatic Biology and Disease
Authors: Shin William S. , Rockson Stanley G.
Published: June 2008

The development of animal model systems for the study of the lymphatic system has resulted in an explosion of information regarding the mechanisms governing lymphatic development and the diseases associated with lymphatic dysfunction. Animal studies have led to a new molecular model of embryonic lymphatic vascular development, and have provided insight into the pathophysiology of both inherited and acquired lymphatic insufficiency. It has become apparent, however, that the importance of the lymphatic system to human disease extends, beyond its role in lymphedema, to many other diverse pathologic processes, including, very notably, inflammation and tumor lymphangiogenesis. Here, we have undertaken a systematic review of the models as they relate to molecular and functional characterization of the development, maturation, genetics, heritable and acquired diseases, and neoplastic implications of the lymphatic system. The translation of these advances into therapies for human diseases associated with lymphatic dysfunction will require the continued study of the lymphatic system through robust animal disease models that simulate their human counterparts.
Lymphatic Vasculature Development
Authors: Oliver Guillermo , Srinivasan R. Sathish
Published: June 2008

The recent identification of lymphatic endothelial cell–specific markers and the generation of mouse models harboring mutations in many genes that encode those markers have enabled us to begin to elucidate the developmental steps that go toward establishing the lymphatic vasculature network. Understanding these basic mechanisms of lymphatic development will improve our ability to diagnose, prevent, and ideally treat lymphatic disorders.
Lymphatic Vessels: Pressure‐ and Flow‐dependent Regulatory Reactions
Authors: Gashev Anatoliy A.
Published: June 2008

This chapter discusses the current status of knowledge on basic pressure/stretch– and flow/shear–dependent regulatory reactions of contracting lymphangions in lymphatic vessels. The balance of passive and active lymph pumps is discussed; the data on pressure measurements in different lymphatic nets of several species are provided. The characteristics of typical changes in lymphatic contractility to the increases in transmural pressures are described and discussed, as is the role of pre‐existing distension in initiation of lymphatic contractions. The up‐to‐date status of knowledge on flow‐dependent adaptive reactions of contracting lymphangions is provided with detailed characterization of the influences of imposed and contraction‐generated flows on lymphatic contractility. The interaction between extrinsic and intrinsic flows in regulation of lymphatic contractility is considered.
Cell Traffic and the Lymphatic Endothelium
Authors: Johnson Louise A. , Jackson David G.
Published: June 2008

The principal immune function of the afferent lymphatics is to bear antigen and leukocytes from peripheral tissues to the draining lymph nodes. Recent research has shown that passage of leukocytes into the afferent lymphatic capillaries is far from an indolent process; rather it is carefully orchestrated by an array of adhesion molecules, as well as by chemokines and their receptors. Here we review the current knowledge of leukocyte trans‐lymphatic endothelial migration and its role in the development of an immune response.
Estimating the Population Burden of Lymphedema
Authors: Rockson Stanley G. , Rivera Kahealani K.
Published: June 2008

Lymphedema is a complex, regional edematous state that ensues when lymph transport is insufficient to maintain tissue homeostasis. The disorder is remarkably prevalent, but the population implications of lymphatic dysfunction are not well‐studied. Prevalence estimates for lymphedema are relatively high, yet its prevalence is likely underestimated. The ability to estimate the burden of disease poses profound implications for current and future lymphedema patients, but the challenge to correctly surmise the incidence and prevalence of lymphedema is complex and the relevant medical literature is scanty. In the absence of the highly desired, prospectively designed and rigorously performed relevant epidemiologic studies, it is instructive to look at the existing studies of lymphedema disease burden. In the current review, the extant literature is examined in the context of the disease setting in which tissue edema is encountered. Incidence or prevalence estimates are provided or inferred, and, where feasible, the size of the subject population is also identified. It is extremely attractive to contemplate that future approaches will entail formal, prospectively designed studies to objectively quantitate incidence and prevalence statistics for individual categories, as well as for the global lymphedema population.
Congenital Lymphatic Malformations
Authors: Blei Francine
Published: June 2008

“Vascular anomalies” represents a spectrum of vascular lesions, of unclear etiology and often with unpredictable behavior. Patients with vascular anomalies represent a unique population, in that they have focal aberrations of vascular development (in vascular malformations) or vascular proliferation (in hemangiomas). The etiology of these disorders is unclear, and likely represents a multifactorial process. Vascular anomalies are an attractive model for the study of human disorders of vasculogenesis (development of the vasculature) and angiogenesis (new vessel growth from existing vessels).