bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of a Cre/loxP recombination system for N-terminal epitope tagging of genes in Tetrahymena
Authors: Busch Clara Jana-Lui , Vogt Alexander , Mochizuki Kazufumi
Published: July 2010

Background

Epitope tagging is a powerful strategy to study the function of proteins. Although tools for C-terminal protein tagging in the ciliated protozoan Tetrahymena thermophila have been developed, N-terminal protein tagging in this organism is still technically demanding.

Results

In this study, we have established a Cre/loxP recombination system in Tetrahymena and have applied this system for the N-terminal epitope tagging of Tetrahymena genes. Cre can be expressed in Tetrahymena and localizes to the macronucleus where it induces precise recombination at two loxP sequences in direct orientation in the Tetrahymena macronuclear chromosome. This Cre/loxP recombination can be used to remove a loxP-flanked drug-resistance marker from an N-terminal tagging construct after it is integrated into the macronucleus.

Conclusions

The system established in this study allows us to express an N-terminal epitope tagged gene from its own endogenous promoter in Tetrahymena.
An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector
Authors: de Backer Marijke WA , Fitzsimons Carlos P , Brans Maike AD , Luijendijk Mieneke CM , Garner Keith M , Vreugdenhil Erno , Adan Roger AH
Published: July 2010

Background

This study compared the transduction efficiencies of an adeno-associated viral (AAV) vector, which was pseudotyped with an AAV1 capsid and encoded the green fluorescent protein (GFP), with a lentiviral (LV) vector, which was pseudotyped with a VSV-G envelop and encoded the discosoma red fluorescent protein (dsRed), to investigate which viral vector transduced the lateral hypothalamus or the amygdala more efficiently. The LV-dsRed and AAV1-GFP vector were mixed and injected into the lateral hypothalamus or into the amygdala of adult rats. The titers that were injected were 1 × 108 or 1 × 109 genomic copies of AAV1-GFP and 1 × 105 transducing units of LV-dsRed.

Results

Immunostaining for GFP and dsRed showed that AAV1-GFP transduced significantly more cells than LV-dsRed in both the lateral hypothalamus and the amygdala. In addition, the number of LV particles that were injected can not easily be increased, while the number of AAV1 particles can be increased easily with a factor 100 to 1000. Both viral vectors appear to predominantly transduce neurons.

Conclusions

This study showed that AAV1 vectors are better tools to overexpress or knockdown genes in the lateral hypothalamus and amygdala of adult rats, since more cells can be transduced with AAV1 than with LV vectors and the titer of AAV1 vectors can easily be increased to transduce the area of interest.
Association of the genetic polymorphisms of the ACE gene and the eNOSgene with lupus nephropathy in northern Chinese population
Authors: Li X , An J , Guo R , Jin Z , Li Y , Zhao Y , Lu F , Lian H , Liu P , Zhao Y , Jin X
Published: July 2010

Background

It has been reported that some single nucleotide polymorphisms (SNPs) of the angiotensin converting enzyme (ACE) gene and the endothelial nitric oxide synthase (eNOS) gene are associated with the development of systemic lupus erythematosus (SLE) and the progression of nephropathy. The aim of this study was to evaluate the possible association between six SNPs (A-5466C,T-3892C,A-240T,C1237T,G2215AandA2350G) of theACEgene and two SNPs (T-786CandG894T) of theeNOSgene with lupus nephropathy in a northern Chinese population.

Methods

In this study, 225 patients with lupus nephropathy were compared to 232 healthy controls, matched by gender, age and ethnicity. Following the extraction of genomic DNA from the leukocytes in the peripheral blood, the genotypes of the eight selected SNPs were determined by the method of PCR-RFLP; the haplotypes were inferred using PHASE 2.1. The associations between the SNPs and the risk of lupus nephropathy were analyzed using Chi-square test and Logistic regression with SPSS13.0 software.

Results

Statistically significant differences of the allele frequency distribution of three SNPs (A-5466C,A2350GandG894T) were observed between cases and controls (P< 0.05). Among the 53 haplotypes identified, the frequencies of five haplotypes (CTTCGA, ACTTAA, ACATGG, ACACGG and ATTCGA) were significantly different between cases and controls (P< 0.05).

Conclusions

Our study indicated an association between the risk of lupus nephropathy and the sequence variations of both theACEgene and theeNOSgene, which may play an important role in the pathogenesis of lupus nephropathy in the northern Chinese population. Further studies are warranted to validate our findings.
Demethylation by 5-aza-2'-deoxycytidine in colorectal cancer cells targets genomic DNA whilst promoter CpG island methylation persists
Authors: Mossman David , Kim Kyu-Tae , Scott Rodney J
Published: July 2010

Background

DNA methylation and histone acetylation are epigenetic modifications that act as regulators of gene expression. Aberrant epigenetic gene silencing in tumours is a frequent event, yet the factors which dictate which genes are targeted for inactivation are unknown. DNA methylation and histone acetylation can be modified with the chemical agents 5-aza-2'-deoxycytidine (5-aza-dC) and Trichostatin A (TSA) respectively. The aim of this study was to analyse de-methylation and re-methylation and its affect on gene expression in colorectal cancer cell lines treated with 5-aza-dC alone and in combination with TSA. We also sought to identify methylation patterns associated with long term reactivation of previously silenced genes.

Method

Colorectal cancer cell lines were treated with 5-aza-dC, with and without TSA, to analyse global methylation decreases by High Performance Liquid Chromatography (HPLC). Re-methylation was observed with removal of drug treatments. Expression arrays identified silenced genes with differing patterns of expression after treatment, such as short term reactivation or long term reactivation. Sodium bisulfite sequencing was performed on the CpG island associated with these genes and expression was verified with real time PCR.

Results

Treatment with 5-aza-dC was found to affect genomic methylation and to a lesser extent gene specific methylation. Reactivated genes which remained expressed 10 days post 5-aza-dC treatment featured hypomethylated CpG sites adjacent to the transcription start site (TSS). In contrast, genes with uniformly hypermethylated CpG islands were only temporarily reactivated.

Conclusion

These results imply that 5-aza-dC induces strong de-methylation of the genome and initiates reactivation of transcriptionally inactive genes, but this does not require gene associated CpG island de-methylation to occur. In addition, for three of our selected genes, hypomethylation at the TSS of an epigenetically silenced gene is associated with the long term reversion of gene expression level brought about by alterations in the epigenetic status following 5-aza-dC treatment.
Late presenters to HIV care and treatment, identification of associated risk factors in HIV-1 infected Indian population
Authors: Mojumdar Kamalika , Vajpayee Madhu , Chauhan Neeraj K , Mendiratta Sanjay
Published: July 2010

Background

Timely access to antiretroviral therapy is a key to controlling HIV infection. Late diagnosis and presentation to care diminish the benefits of antiretrovirals and increase risk of transmission. We aimed to identify late presenters in patients sent for first CD4 T cell count after HIV diagnosis, for therapy initiation evaluation. Further we aimed at identifying patient factors associated with higher risk of late presentation.

Methods

Retrospective data collection and analysis was done for 3680 subjects visiting the laboratory for CD4 T cell counts between 2001 and 2007. We segregated the patients on basis of their CD4 T cell counts after first HIV diagnosis. Factors associated with risk of late presentation to CD4 T cell counts after HIV diagnosis were identified using univariate analysis, and the strength of association of individual factor was assessed by calculation of odds ratios.

Results

Of 3680 subjects, 2936 (83.37%) were defined as late presenters. Late testing varied among age groups, transmission categories, and gender. Males were twice as likely to present late as compared to females. We found significant positive association of heterosexual transmission route (p < 0.001), and older age groups of 45 years and above (p = 0.0004) to late presentation. Female sex, children below 14 years of age and sexual contact with HIV positive spouse were associated with significantly lower risks to presenting late. Intravenous drug users were also associated with lower risks of late presentation, in comparison to heterosexual transmission route.

Conclusions

The study identifies HIV infected population groups at a higher risk of late presentation to care and treatment. The risk factors identified to be associated with late presentation should be utilised in formulating targeted public health interventions in order to improve early HIV diagnosis.
A randomised, multi-centre, prospective, double blind pilot-study to evaluate safety and efficacy of the non-absorbable Optilene® Mesh Elastic versus the partly absorbable Ultrapro® Mesh for incisional hernia repair
Authors: Seiler Christoph , Baumann Petra , Kienle Peter , Kuthe Andreas , Kuhlgatz Jens , Engemann Rainer , v Frankenberg Moritz , Knaebel Hanns-Peter
Published: July 2010

Background

Randomised controlled trials with a long term follow-up (3 to 10 years) have demonstrated that mesh repair is superior to suture closure of incisional hernia with lower recurrence rates (5 to 20% versus 20 to 63%). Yet, the ideal size and material of the mesh are not defined. So far, there are few prospective studies that evaluate the influence of the mesh texture on patient's satisfaction, recurrence and complication rate. The aim of this study is to evaluate, if a non-absorbable mesh (Optilene®Mesh Elastic) will result in better health outcomes compared to a partly absorbable mesh (Ultrapro®Mesh).

Methods/Design

In this prospective, randomised, double blind study, eighty patients with incisional hernia after a midline laparotomy will be included. Primary objective of this study is to investigate differences in the physical functioning score from the SF-36 questionnaire 21 days after mesh insertion. Secondary objectives include the evaluation of the patients' daily activity, pain, wound complication and other surgical complications (hematomas, seromas), and safety within six months after intervention.

Discussion

This study investigates mainly from the patient perspective differences between meshes for treatment of incisional hernias. Whether partly absorbable meshes improve quality of life better than non-absorbable meshes is unclear and therefore, this trial will generate further evidence for a better treatment of patients.

Trial registration

NCT00646334
SOX11 expression correlates to promoter methylation and regulates tumor growth in hematopoietic malignancies
Authors: Gustavsson Elin , Sernbo Sandra , Andersson Elin , Brennan Donal J. , Dictor Michael , Jerkeman Mats , Borrebaeck Carl A. K. , Ek Sara
Published: July 2010

Background

The transcription factor SOX11 plays an important role in embryonic development of the central nervous system (CNS) and is expressed in the adult immature neuron but is normally not expressed in any other adult tissue. It has recently been reported to be implicated in various malignant neoplasms, including several lymphoproliferative diseases, by its specific expression and in some cases correlation to prognosis. SOX11 has been shown to prevent gliomagenesis in vivo but the causes and consequences of aberrant expression of SOX11 outside the CNS remain unexplained.

Results

We now show the first function of SOX11 in lymphoproliferative diseases, by demonstrating in vitro its direct involvement in growth regulation, as assessed by siRNA-mediated silencing and ectopic overexpression in hematopoietic malignancies. Gene Chip analysis identified cell cycle regulatory pathways, including Rb-E2F, to be associated with SOX11-induced growth reduction. Furthermore, promoter analysis revealed that SOX11 is silenced through DNA methylation in B cell lymphomas, suggesting that its regulation is epigenetically controlled.

Conclusions

The data show that SOX11 is not a bystander but an active and central regulator of cellular growth, as both siRNA-mediated knock-down and ectopic overexpression of SOX11 resulted in altered proliferation. Thus, these data demonstrate a tumor suppressor function for SOX11 in hematopoietic malignancies and revealed a potential epigenetic regulation of this developmentally involved gene.
Extensive analysis of D7S486 in primary gastric cancer supports TESTIN as a candidate tumor suppressor gene
Authors: Ma Haiqing , Weng Desheng , Chen Yibing , Huang Wei , Pan Ke , Wang Hui , Sun Jiancong , Wang Qijing , Zhou Zhiwei , Wang Huiyun , Xia Jianchuan
Published: July 2010

Background

High frequency of loss of heterozygosity (LOH) was found at D7S486 in primary gastric cancer (GC). And we found a high frequency of LOH region on 7q31 in primary GC from China, and identified D7S486 to be the most frequent LOH locus. This study was aimed to determine what genes were affected by the LOH and served as tumor suppressor genes (TSGs) in this region. Here, a high-throughput single nucleotide polymorphisms (SNPs) microarray fabricated in-house was used to analyze the LOH status around D7S486 on 7q31 in 75 patients with primary GC. Western blot, immunohistochemistry, and RT-PCR were used to assess the protein and mRNA expression of TESTIN (TES) in 50 and 140 primary GC samples, respectively. MTS assay was used to investigate the effect of TES overexpression on the proliferation of GC cell lines. Mutation and methylation analysis were performed to explore possible mechanisms of TES inactivation in GC.

Results

LOH analysis discovered five candidate genes (ST7,FOXP2,MDFIC,TESandCAV1) whose frequencies of LOH were higher than 30%. However, onlyTESshowed the potential to be a TSG associated with GC. Among 140 pairs of GC samples, decreasedTESmRNA level was found in 96 (68.6%) tumor tissues when compared with matched non-tumor tissues (p< 0.001). Also, reduced TES protein level was detected in 36 (72.0%) of all 50 tumor tissues by Western blot (p= 0.001). In addition, immunohistochemical staining result was in agreement with that of RT-PCR and Western blot. Down regulation of TES was shown to be correlated with tumor differentiation (p= 0.035) and prognosis (p= 0.035, log-rank test). Its overexpression inhibited the growth of three GC cell lines. Hypermethylation ofTESpromoter was a frequent event in primary GC and GC cell lines. However, no specific gene mutation was observed in the coding region of theTESgene.

Conclusions

Collectively, all results support the role ofTESas a TSG in gastric carcinogenesis and thatTESis inactivated primarily by LOH and CpG island methylation.
Proteomic analysis of differentially expressed proteins in Penaeus monodon hemocytes after Vibrio harveyi infection
Authors: Somboonwiwat Kunlaya , Chaikeeratisak Vorrapon , Wang Hao-Ching , Fang Lo Chu , Tassanakajon Anchalee
Published: July 2010

Background

Viral and bacterial diseases can cause mass mortalities in commercial shrimp aquaculture. In contrast to studies on the antiviral response, the responses of shrimps to bacterial infections by high throughput techniques have been reported only at the transcriptional level and not at the translational level. In this study, a proteomic analysis of shrimp hemocytes to identify differentially expressed proteins in response to a luminous bacterium Vibrio harveyi was evaluated for its feasibility and is reported for the first time.

Results

The two-dimensional gel electrophoresis (2-DE) patterns of the hemocyte proteins from the unchallenged and V. harveyi challenged shrimp, Penaeus monodon, at 24 and 48 h post infection were compared. From this, 27 differentially expressed protein spots, and a further 12 weakly to non-differentially regulated control spots, were selected for further analyses by the LC-ESI-MS/MS. The 21 differentially expressed proteins that could be identified by homologous annotation were comprised of proteins that are directly involved in the host defense responses, such as hemocyanin, prophenoloxidase, serine proteinase-like protein, heat shock protein 90 and alpha-2-macroglobulin, and those involved in signal transduction, such as the14-3-3 protein epsilon and calmodulin. Western blot analysis confirmed the up-regulation of hemocyanin expression upon bacterial infection. The expression of the selected proteins which were the representatives of the down-regulated proteins (the 14-3-3 protein epsilon and alpha-2-macroglobulin) and of the up-regulated proteins (hemocyanin) was further assessed at the transcription level using real-time RT-PCR.

Conclusions

This work suggests the usefulness of a proteomic approach to the study of shrimp immunity and revealed hemocyte proteins whose expression were up regulated upon V. harveyi infection such as hemocyanin, arginine kinase and down regulated such as alpha-2-macroglobulin, calmodulin and 14-3-3 protein epsilon. The information is useful for understanding the immune system of shrimp against pathogenic bacteria.
Reliability of 95% confidence interval revealed by expected quality-of-life scores: an example of nasopharyngeal carcinoma patients after radiotherapy using EORTC QLQ-C 30
Authors: Chien Tsair-Wei , Lin Shun-Jin , Wang Wen-Chung , Leung Henry WC , Lai Wen-Pin , Chan Agnes LF
Published: July 2010

Background

Many researchers use observed questionnaire scores to evaluate score reliability and to make conclusions and inferences regarding quality-of-life outcomes. The amount of false alarms from medical diagnoses that would be avoided if observed scores were substituted with expected scores is interesting, and understanding these differences is important for the care of cancer patients. Using expected scores to estimate the reliability of 95% confidence intervals (CIs) is rarely reported in published papers. We investigated the reliability of patient responses to a quality-of-life questionnaire and made recommendations for future studies of the quality of life of patients.

Methods

A total of 115 patients completed the EORTC core questionnaire QLQ-C30 (version 3) after radiotherapy. The observed response scores, assumed to be one-dimensional, were summed and transformed into expected scores using the Rasch rating scale model with WINSTEPS software. A series of simulations was performed using a unified bootstrap procedure after manipulating scenarios with different questionnaire lengths and patient numbers to estimate the reliability at 95% confidence intervals. Skewness analyses of the 95% CIs were compared to detect different effects between groups according to the two data sets of observed and expected response scores.

Results

We found that (1) it is necessary to report CIs for reliability and skewness coefficients in papers; (2) data derived from expected response scores are preferable to making inferences; and (3) visual representations displaying the 95% CIs of skewness values applied to item-by-item analyses can provide a useful interpretation of quality-of-life outcomes.

Conclusion

Reliability coefficients can be reported with 95% CIs by statistical software to evaluate the internal consistency of respondent scores on questionnaire items. The SPSS syntax procedures for estimating the reliability of the 95% CI, expected score generation and visual skewness analyses are demonstrated in this study. We recommend that effect sizes such as a 95% CI be reported along with p values reporting significant differences in quality-of-life studies.
Solitary colonic metastasis from renal cell carcinoma presenting as a surgical emergency nine years post-nephrectomy
Authors: Jadav Alka M , Thrumurthy Sri G , DeSousa Bernard A
Published: July 2010

Late colonic metastasis following curative surgery for renal cell carcinoma has rarely been described. We present the first reported case of solitary colonic renal cell carcinoma metastasis presenting as an intra-abdominal bleed, nine years post-nephrectomy.
Proto-oncogene c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways
Authors: Li-Ping Zheng , Da-Lei Zhang , Jian Huang , Liang-Quan Xu , Ai-Xia Xu , Xiao-Yu Du , Dan-Feng Tang , Yue-Hui Zheng
Published: July 2010

Background

c-erbB2, a proto-oncogene coding epidermal growth factor receptor-like receptor, also as a chemosensitivity/prognosis marker for gynecologic cancer, may be involved in initiation of growth of rat primordial follicles. The aim of the present study is to investigate the role and signal pathway of c-erbB2 in onset of rat primordial follicle development.

Methods

The expression of c-erbB2 mRNA and protein in neonatal ovaries cultured 4 and 8 days with/without epidermal growth factor (EGF) were examined by in situ hybridization, RT-PCR and western blot. The function of c-erbB2 in the primordial folliculogenesis was abolished by small interfering RNA transfection. Furthermore, MAPK inhibitor PD98059 and PKC inhibitor calphostin were used to explore the possible signaling pathway of c-erbB2 in primordial folliculogenesis.

Results

The results showed that c-erbB2 mRNA was expressed in ooplasm and the expression of c-erbB2 decreased after transfection with c-erbB2 siRNA. Treatment with EGF at 50 ng/ml significantly increased c-erbB2 expression and primary and secondary follicle formation in ovaries. However, this augmenting effect was remarkably inhibited by c-erbB2 siRNA transfection. Furthermore, folliculogenesis offset was blocked by calphostin (5 × 10(-4) mmol/L) and PD98059 (5 × 10(-2) mmol/L), but both did not down-regulate c-erbB2 expression. In contrast, the expressions of p-ERK and p-PKC were decreased obviously by c-erbB2 siRNA transfection.

Conclusions

c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways, suggesting an important role of c-erbB2 in rat primordial follicle initiation and development.
Serious limitations of the QTL/Microarray approach for QTL gene discovery
Authors: Verdugo Ricardo A , Farber Charles R , Warden Craig H , Medrano Juan F
Published: July 2010

Background

It has been proposed that the use of gene expression microarrays in nonrecombinant parental or congenic strains can accelerate the process of isolating individual genes underlying quantitative trait loci (QTL). However, the effectiveness of this approach has not been assessed.

Results

Thirty-seven studies that have implemented the QTL/microarray approach in rodents were reviewed. About 30% of studies showed enrichment for QTL candidates, mostly in comparisons between congenic and background strains. Three studies led to the identification of an underlying QTL gene. To complement the literature results, a microarray experiment was performed using three mouse congenic strains isolating the effects of at least 25 biometric QTL. Results show that genes in the congenic donor regions were preferentially selected. However, within donor regions, the distribution of differentially expressed genes was homogeneous once gene density was accounted for. Genes within identical-by-descent (IBD) regions were less likely to be differentially expressed in chromosome 2, but not in chromosomes 11 and 17. Furthermore, expression of QTL regulated in cis (cis eQTL) showed higher expression in the background genotype, which was partially explained by the presence of single nucleotide polymorphisms (SNP).

Conclusions

The literature shows limited successes from the QTL/microarray approach to identify QTL genes. Our own results from microarray profiling of three congenic strains revealed a strong tendency to select cis-eQTL over trans-eQTL. IBD regions had little effect on rate of differential expression, and we provide several reasons why IBD should not be used to discard eQTL candidates. In addition, mismatch probes produced false cis-eQTL that could not be completely removed with the current strains genotypes and low probe density microarrays. The reviewed studies did not account for lack of coverage from the platforms used and therefore removed genes that were not tested. Together, our results explain the tendency to report QTL candidates as differentially expressed and indicate that the utility of the QTL/microarray as currently implemented is limited. Alternatives are proposed that make use of microarray data from multiple experiments to overcome the outlined limitations.
Stem cell proliferation during in vitro development of the model cestode Mesocestoides corti from larva to adult worm
Authors: Koziol Uriel , Domínguez María F , Marín Mónica , Kun Alejandra , Castillo Estela
Published: July 2010

Background

In free-living flatworms somatic differentiated cells do not divide, and a separate population of stem cells (called neoblasts) is responsible for cell proliferation and renewal. In cestodes, there is evidence that similar mechanisms of cell renewal exist.

Results

In this work, we have characterized proliferative cells during the development of the model cestode Mesocestoides corti from larva (tetrathyridium) to young segmented worm. This was done by two complementary strategies with congruent results: characterizing cells in S phase and their progeny by incorporation of 5-bromo-2'-deoxyuridine, and characterizing cells in M phase by arresting mitotic cells with colchicine and studying their morphology and distribution. Proliferative cells are localized only in the inner parenchyma, particularly in close proximity to the inner muscle layer, but not in the cortical parenchyma nor in the sub-tegumental tissue. After proliferation some of these cells migrate to the outer regions were they differentiate. In the larvae, proliferative cells are more abundant in the anterior regions (scolex and neck), and their number diminishes in an antero-posterior way. During the development of adult segments periodic accumulation of proliferative cells are observed, including a central mass of cells that constitutes the genital primordium, which grows at least in part due to in situ proliferation. In later segments, the inner cells of genital primordia cease to proliferate and adopt a compact distribution, and proliferative cells are also found in the testes primordia.

Conclusions

Proliferative cells have a characteristic localization and morphology throughout development from larva to adult of Mesocestoides corti, which is similar, and probably evolutionary conserved, to that described in other model cestodes. The characteristics of proliferative cells suggest that these consist of undifferentiated stem cells.
The timing of perinatal hypoxia/ischemia events in term neonates: a retrospective autopsy study. HSPs, ORP-150 and COX2 are reliable markers to classify acute, perinatal events
Authors: Riezzo Irene , Neri Margherita , De Stefano Francesco , Fulcheri Ezio , Ventura Francesco , Pomara Cristoforo , Rabozzi Roberto , Turillazzi Emanuela , Fineschi Vittorio
Published: July 2010

Background

The understanding of the cellular responses implicated in perinatal brain damages and the characterization of the various mechanisms involved might open new horizons for understanding the time of onset of a brain hypoxic-ischemic lesion and for effective therapeutic strategies.

Methods

We performed an immunohistochemical investigation on brain and brainstem sections of 47 peripartum deaths. The gradation and localization of the expression of antibodies such as TNFα, IL-1β, IL-6, HSPs, β APP, anti-TrypH, GAP43, GFAP, COX2, ORP-150, could be correlated with an hypoxic-ischemic damage to document a significant correlation between response and the time of onset acute (≤8 hs) or non-acute (≥8 hs ≤48 hs).

Results and Discussions

In non-acute cases HSP70 reaction was prominent in the neuron cytoplasm, while in acute cases a mild reaction was evident in sporadic fields. HSP90 exhibited a similar pattern of positivity as HSP70. In acute group, ORP150 expressed an intense reaction showing a granular pattern in the cytoplasm of the neurons in the cortex of the infarcted areas. In non-acute group the positive reaction was more intense in astrocytes and less extended in neurons. COX2 reaction exhibited the strongest positive reaction in the neuronal cell bodies of acute cases, while a immunolabeling was prominent in the glial cytoplasm in the non-acute cases.

Conclusions

Chaperones HSP70 and 90, ORP-150 reaction, and COX2 protein, have provided very interesting results. These results would suggest to the clinicians to extend the differential diagnosis of a too large perinatal hypoxic-ischemic insult category to delineate a more accurate chronological judgement.
The instantaneous helical axis of the subtalar and talocrural joints: a non‐invasive in vivo dynamic study
Authors: Sheehan Frances T
Published: July 2010

Background

An understanding of rear‐foot (talocrural and subtalar joints) kinematics is critical for diagnosing foot pathologies, designing total ankle implants, treating rear‐foot injuries and quantifying gait abnormalities. The majority of kinematic data available have been acquired through static cadaver work or passive in vivo studies. The applicability of these data to dynamic in vivo situations remains unknown. Thus, the purpose of this study was to fully quantify subtalar, talocrural and calcaneal‐tibial in vivo kinematics in terms of the instantaneous helical axis (IHA) in twenty‐five healthy ankles during a volitional activity that simulated single‐leg toe‐raises with partial‐weight support, requiring active muscle control.

Methods

Subjects were each placed supine in a 1.5 T MRI and asked to repeat this simulated toe‐raise while a full sagittal‐cine‐phase contrast (dynamic) MRI dataset was acquired. From the cine‐phase contrast velocity a full kinematic description for each joint was derived.

Results

Nearly all motion quantified at the calcaneal‐tibial joint was attributable to the talocrural joint. The subtalar IHA orientation and position were highly variable; whereas, the talocrural IHA orientation and position were extremely consistent.

Conclusion

The talocrural was well described by the IHA and could be modeled as a fixed‐hinge joint, whereas the subtalar could not be.
Low cholesterol? Don't brag yet … hypocholesterolemia blunts HAART effectiveness: a longitudinal study
Authors: Míguez María Jose , Lewis John E , Bryant Vaughn E , Rosenberg Rhonda , Burbano Ximena , Fishman Joel , Asthana Deshratn , Duan Rui , Madhavan Nair , Malow Robert M
Published: July 2010

Background

In vitro studies suggest that reducing cholesterol inhibits HIV replication. However, this effect may not hold in vivo, where other factors, such as cholesterol's immunomodulatory properties, may interact.

Methods

Fasting blood samples were obtained on 165 people living with HIV at baseline and after 24 weeks on highly active antiretroviral therapy (HAART). Participants were classified as hypocholesterolemic (HypoCHL; <150 mg/dl) or non‐HypoCHL (>150 mg/dl) and were compared on viro‐immune outcomes.

Results

At baseline, participants with HypoCHL (40%) exhibited lower CD4 (197 ± 181 vs. 295 ± 191 cells/mm3, p = 0.02) and CD8 (823 ± 448 vs. 1194 ± 598 cells/mm3, p = 0.001) counts and were more likely to have detectable viral loads (OR = 3.5, p = 0.01) than non‐HypoCHL controls. After HAART, participants with HypoCHL were twice as likely to experience a virological failure >400 copies (95% CI 1‐2.6, p = 0.05) and to exhibit <200 CD4 (95% CI 1.03‐2.9, p = 0.04) compared with non‐HypoCHL. Low thymic output was related to poorer CD4 cell response in HypoCHL subjects. Analyses suggest a dose‐response relationship with every increase of 50 mg/dl in cholesterol related to a parallel rise of 50 CD4 cells.

Conclusions

The study implicates, for the first time, HypoCHL with impaired HAART effectiveness, including limited CD4 repletion by the thymus and suboptimal viral clearance.
Pharmacoproteomic study of the effects of chondroitin and glucosamine sulfate on human articular chondrocytes
Authors: Calamia Valentina , Ruiz-Romero Cristina , Rocha Beatriz , Fernández-Puente Patricia , Mateos Jesús , Montell Eulàlia , Vergés Josep , Blanco Francisco J
Published: July 2010

Introduction

Chondroitin sulfate (CS) and glucosamine sulfate (GS) are symptomatic slow-acting drugs for osteoarthritis (OA) widely used in clinic. Despite their widespread use, knowledge of the specific molecular mechanisms of their action is limited. The aim of this work is to explore the utility of a pharmacoproteomic approach for the identification of specific molecules involved in the pharmacological effect of GS and CS.

Methods

Chondrocytes obtained from three healthy donors were treated with GS 10 mM and/or CS 200 μg/mL, and then stimulated with interleukin-1β (IL-1β) 10 ng/mL. Whole cell proteins were isolated 24 hours later and resolved by two-dimensional electrophoresis. The gels were stained with SYPRORuby. Modulated proteins were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF/TOF) mass spectrometry. Real-time PCR and Western blot analyses were performed to validate our results.

Results

A total of 31 different proteins were altered by GS or/and CS treatment when compared to control. Regarding their predicted biological function, 35% of the proteins modulated by GS are involved in signal transduction pathways, 15% in redox and stress response, and 25% in protein synthesis and folding processes. Interestingly, CS affects mainly energy production (31%) and metabolic pathways (13%), decreasing the expression levels of ten proteins. The chaperone GRP78 was found to be remarkably increased by GS alone and in combination with CS, a fact that unveils a putative mechanism for the reported anti-inflammatory effect of GS in OA. On the other hand, the antioxidant enzyme superoxide dismutase 2 (SOD2) was significantly decreased by both drugs and synergistically by their combination, thus suggesting a drug-induced decrease of the oxidative stress caused by IL-1β in chondrocytes.

Conclusions

CS and GS differentially modulate the proteomic profile of human chondrocytes. This pharmacoproteomic approach unravels the complex intracellular mechanisms that are modulated by these drugs on IL1β-stimulated human articular chondrocytes.
Differential effects of locally and systemically administered soluble glycoprotein 130 on pain and inflammation in experimental arthritis
Authors: Boettger Michael K , Leuchtweis Johannes , Kümmel Diana , Gajda Mieczyslaw , Bräuer Rolf , Schaible Hans-Georg
Published: July 2010

Introduction

Interleukin-6 (IL-6) is a key player in systemic arthritis, involved in inflammation and joint destruction. IL-6 signalling has also been revealed in nerve cells. Recently, IL-6 and in particular IL-6 together with its soluble IL-6 receptor (sIL-6R) were shown to induce a long-lasting robust sensitization of joint nociceptors for mechanical stimuli which was difficult to reverse, suggesting that IL-6 signalling plays a significant role in the generation and maintenance of arthritic pain. Here we tested in a preclinical model of arthritis, antigen-induced arthritis (AIA) in the rat, whether systemic or local neutralization of IL-6/sIL-6R complexes with soluble glycoprotein 130 (sgp130) alters arthritic pain and how sgp130 influences the inflammatory process in AIA.

Methods

Rats with AIA were either treated with sgp130 or saline intra-peritoneally or intra-articularly (each group n = 9). Then, pain-related and locomotor behaviour, as well as joint swelling, were measured during an observation period of 21 days, followed by histopathological end-point analysis for inflammatory and destructive changes.

Results

A single intra-articular application of sgp130 at the time of AIA induction barely reduced the development of AIA, but significantly attenuated pain-related behaviour, that is, primary mechanical hyperalgesia in the acute phase of AIA. By contrast, repeated systemic application of sgp130 after onset of AIA only slightly attenuated pain at a late stage of AIA. None of the treatments reduced secondary hyperalgesia. Furthermore, in the present study joint destruction at 21 days was significantly attenuated after intra-articular sgp130 treatment, but not after systemic sgp130.

Conclusions

In addition to its role in chronic inflammation, IL-6 in the joint plays a significant role in the generation and maintenance of arthritic joint pain at acute and chronic stages of AIA. The particular effectiveness of intra-articular injection of sgp130 indicates, first, that IL-6/sIL-6R in the inflamed joint, rather than circulating IL-6/sIL-6R, is responsible for the generation of hyperalgesia, and, second, that early neutralization of IL-6/sIL-6R is particularly successful in producing antinociception. Furthermore, neutralization of IL-6/sIL-6R (and possibly other cytokines which use the transmembrane signal-transducing subunit gp130) directly at the site of joint inflammation seems to be effective in the prevention of joint destruction.
Phase III Study Comparing Second- and Third-Generation Regimens With Concurrent Thoracic Radiotherapy in Patients With Unresectable Stage III Non–Small-Cell Lung Cancer: West Japan Thoracic Oncology Group WJTOG0105
Authors: Yamamoto Nobuyuki , Nakagawa Kazuhiko , Nishimura Yasumasa , Tsujino Kayoko , Satouchi Miyako , Kudo Shinzoh , Hida Toyoaki , Kawahara Masaaki , Takeda Koji , Katakami Nobuyuki , Sawa Toshiyuki , Yokota Soichiro , Seto Takashi , Imamura Fumio , Saka Hideo , Iwamoto Yasuo , Semba Hiroshi , Chiba Yasutaka , Uejima Hisao , Fukuoka Masahiro
Published: July 2010

Purpose

This phase III trial of concurrent thoracic radiotherapy (TRT) was conducted to compare third-generation chemotherapy with second-generation chemotherapy in patients with unresectable stage III non–small-cell lung cancer (NSCLC).

Patients and Methods

Eligible patients received the following treatments: A (control), four cycles of mitomycin (8 mg/m2 on day 1)/vindesine (3 mg/m2 on days 1, 8)/cisplatin (80 mg/m2 on day 1) plus TRT 60 Gy (treatment break for 1 week); B, weekly irinotecan (20 mg/m2)/carboplatin (area under the plasma concentration-time curve [AUC] 2) for 6 weeks plus TRT 60 Gy, followed by two courses of irinotecan (50 mg/m2 on days1, 8)/carboplatin (AUC 5 on day1); C, weekly paclitaxel (40 mg/m2)/carboplatin (AUC 2) for 6 weeks plus TRT 60 Gy, followed by two courses of paclitaxel (200 mg/m2 on day1)/carboplatin (AUC 5 on day 1).

Results

The median survival time and 5-year survival rates were 20.5, 19.8, and 22.0 months and 17.5%, 17.8%, and 19.8% in arms A, B, and C, respectively. Although no significant differences in overall survival were apparent among the treatment arms, noninferiority of the experimental arms was not achieved. The incidences of grade 3 to 4 neutropenia, febrile neutropenia, and gastrointestinal disorder were significantly higher in arm A than in arm B or C (P < .001). Chemotherapy interruptions were more common in arm B than in arm A or C.

Conclusion

Arm C was equally efficacious and exhibited a more favorable toxicity profile among three arms. Arm C should be considered a standard regimen in the management of locally advanced unresectable NSCLC.
American Society of Clinical Oncology Clinical Practice Guideline: Update on Adjuvant Endocrine Therapy for Women With Hormone Receptor–Positive Breast Cancer
Authors: Burstein Harold J. , Prestrud Ann Alexis , Seidenfeld Jerome , Anderson Holly , Buchholz Thomas A. , Davidson Nancy E. , Gelmon Karen E. , Giordano Sharon H. , Hudis Clifford A. , Malin Jennifer , Mamounas Eleftherios P. , Rowden Diana , Solky Alexander J. , Sowers MaryFran R. , Stearns Vered , Winer Eric P. , Somerfield Mark R. , Griggs Jennifer J.
Published: July 2010

Purpose

To develop evidence-based guidelines, based on a systematic review, for endocrine therapy for postmenopausal women with hormone receptor–positive breast cancer.

Methods

A literature search identified relevant randomized trials. Databases searched included MEDLINE, PREMEDLINE, the Cochrane Collaboration Library, and those for the Annual Meetings of the American Society of Clinical Oncology (ASCO) and the San Antonio Breast Cancer Symposium (SABCS). The primary outcomes of interest were disease-free survival, overall survival, and time to contralateral breast cancer. Secondary outcomes included adverse events and quality of life. An expert panel reviewed the literature, especially 12 major trials, and developed updated recommendations.

Results

An adjuvant treatment strategy incorporating an aromatase inhibitor (AI) as primary (initial endocrine therapy), sequential (using both tamoxifen and an AI in either order), or extended (AI after 5 years of tamoxifen) therapy reduces the risk of breast cancer recurrence compared with 5 years of tamoxifen alone. Data suggest that including an AI as primary monotherapy or as sequential treatment after 2 to 3 years of tamoxifen yields similar outcomes. Tamoxifen and AIs differ in their adverse effect profiles, and these differences may inform treatment preferences.

Conclusion

The Update Committee recommends that postmenopausal women with hormone receptor–positive breast cancer consider incorporating AI therapy at some point during adjuvant treatment, either as up-front therapy or as sequential treatment after tamoxifen. The optimal timing and duration of endocrine treatment remain unresolved. The Update Committee supports careful consideration of adverse effect profiles and patient preferences in deciding whether and when to incorporate AI therapy.
Deletion of thePER3Gene on Chromosome 1p36 in Recurrent ER-Positive Breast Cancer
Authors: Climent Joan , Perez-Losada Jesus , Quigley David A. , Kim Il-Jin , Delrosario Reyno , Jen Kuang-Yu , Bosch Ana , Lluch Ana , Mao Jian-Hua , Balmain Allan
Published: July 2010

Purpose

To investigate the role of the PER3 circadian rhythm gene, located within the commonly deleted region of chromosome 1p36, in human breast cancer development.

Patients and Methods

The frequency of genetic alterations at 1p36 and PER3 gene copy number status were analyzed in 180 lymph node–negative breast cancers from patients who had received treatment with chemotherapy and/or tamoxifen. The expression levels of PER3 were also analyzed using published microarray profiles from > 400 breast cancer samples. Finally, the effect of loss of Per3 on tumor susceptibility was tested using two mouse models of breast cancer.

Results

Deletion of PER3 is directly related to tumor recurrence in patients with estrogen receptor (ER) – positive breast cancers treated with tamoxifen. Low expression of PER3 mRNA is associated with poor prognosis, particularly in a subset of tumors that are ER positive, and either luminal A or ERBB2-positive tumors. Mice deficient in Per3 showed increased susceptibility to breast cancer induced by carcinogen treatment or by overexpression of Erbb2.

Conclusion

Disruption of PER3 function may serve as an indicator of probability of tumor recurrence in patients with ER-positive tumors. Further investigations of this pathway may reveal links between deregulation of sleep homeostasis and breast tumorigenesis.
Siltuximab, a Novel Anti–Interleukin-6 Monoclonal Antibody, for Castleman's Disease
Authors: van Rhee Frits , Fayad Luis , Voorhees Peter , Furman Richard , Lonial Sagar , Borghaei Hossein , Sokol Lubomir , Crawford Julie , Cornfeld Mark , Qi Ming , Qin Xiang , Herring Jennifer , Casper Corey , Kurzrock Razelle
Published: July 2010

Purpose

Interleukin-6 (IL-6) has emerged as a key factor in the pathogenesis of the atypical lymphoproliferative disorder Castleman's disease (CD). Siltuximab is a new anti–IL-6, chimeric monoclonal antibody with potential therapeutic benefit in patients with CD.

Methods

We report interim results from an open-label, dose-finding, seven-cohort, phase I study in which patients with symptomatic, multicentric or unresectable, unicentric CD received siltuximab at 1-, 2-, or 3-week intervals. The main efficacy end point of clinical benefit response (CBR) was defined as a composite of clinical and laboratory measures relevant to the management of CD. In addition, radiologic response was independently assessed by using modified Cheson criteria.

Results

Eighteen (78%) of 23 patients (95% CI, 56% to 93%) achieved CBR, and 12 patients (52%) demonstrated objective tumor response. All 11 patients (95% CI, 72% to 100%) treated with the highest dose of 12 mg/kg achieved CBR, and eight patients (73%) achieved objective tumor response. Overall objective-response duration ranged from 44 to ≥ 889 days, and one patient had complete response for ≥ 318 days. Hemoglobin increased markedly in 19 patients (median increase, 2.1 g/dL; range, 0.2 to 4.7 g/dL) in the absence of transfusion or erythropoiesis-stimulating agents. No dose-limiting toxicity was reported, and only three patients had grade 3 or higher adverse events after a median exposure of 331 days (range, 1 to 1,148 days).

Conclusion

These interim results strongly suggest that siltuximab is an effective treatment with favorable safety for the management of CD. An additional study is planned to fully evaluate safety and efficacy at the recommended dose of 12 mg/kg every 3 weeks.
Is Mammographic Breast Density a Breast Cancer Risk Factor in Women With BRCA Mutations?
Authors: Passaperuma Kavitha , Warner Ellen , Hill Kimberley A. , Gunasekara Anoma , Yaffe Martin J.
Published: July 2010

Purpose

Increased mammographic breast density is well recognized as a breast cancer risk factor in the general population. However, it is unclear whether it is a risk factor in women with BRCA mutations. We present the results of a nested case-control screening study investigating the relationship between breast density and breast cancer incidence in this population.

Patients and Methods

Women ages 25 to 65 years with known BRCA mutations were enrolled onto a single-center, high-risk breast cancer screening program. Using a computer-aided technique (Cumulus), quantitative percentage density (PD) was measured for each participant on her baseline mammogram by a single, blinded observer.

Results

Between November 1997 and March 2008, 462 women (mean age, 44 years; 245 BRCA1 and 217 BRCA2) were screened and 50 breast cancers were diagnosed (38 invasive, 12 ductal carcinoma in situ [DCIS]). Density was not measured in 40 women of whom four developed cancer (three invasive, one DCIS). Mean PD (± standard deviation [SD]) for 376 women who did not develop breast cancer was 34% (23) compared with 31% (21) for 46 women with cancer (P = .51). Logistic regression model of breast cancer incidence and PD revealed an odds ratio of 0.99 (± 0.01 SD) for a one-unit increase in PD (P = .44). Results remained nonsignificant in multivariate analysis, as well as when women with pure DCIS were excluded.

Conclusion

Increased mammographic breast density is not associated with higher breast cancer incidence in women with BRCA mutations. On the basis of these findings, density should not be considered a factor for these women in decision making regarding prophylactic surgery or chemoprevention.