bluuebunny/crossref_metadata_2025 huggingface dataset:
Metagenomic islands of hyperhalophiles: the case of Salinibacter ruber
Authors: Pašić Lejla , Rodriguez-Mueller Beltran , Martin-Cuadrado Ana-Belen , Mira Alex , Rohwer Forest , Rodriguez-Valera Francisco
Published: December 2009

Background

Saturated brines are extreme environments of low diversity.Salinibacter ruberis the only bacterium that inhabits this environment in significant numbers. In order to establish the extent of genetic diversity in natural populations of this microbe, the genomic sequence of reference strain DSM 13855 was compared to metagenomic fragments recovered from climax saltern crystallizers and obtained with 454 sequencing technology. This kind of analysis reveals the presence of metagenomic islands, i.e. highly variable regions among the different lineages in the population.

Results

Three regions of the sequenced isolate were scarcely represented in the metagenome thus appearing to vary among co-occurringS. rubercells. These metagenomic islands showed evidence of extensive genomic corruption with atypically low GC content, low coding density, high numbers of pseudogenes and short hypothetical proteins. A detailed analysis of island gene content showed that the genes in metagenomic island 1 code for cell surface polysaccharides. The strain-specific genes of metagenomic island 2 were found to be involved in biosynthesis of cell wall polysaccharide components. Finally, metagenomic island 3 was rich in DNA related enzymes.

Conclusion

The genomic organisation ofS. rubervariable genomic regions showed a number of convergences with genomic islands of marine microbes studied, being largely involved in variable cell surface traits. This variation at the level of cell envelopes in an environment devoid of grazing pressure probably reflects a global strategy of bacteria to escape phage predation.
Computational prediction of essential genes in an unculturable endosymbiotic bacterium, Wolbachia of Brugia malayi
Authors: Holman Alexander G , Davis Paul J , Foster Jeremy M , Carlow Clotilde KS , Kumar Sanjay
Published: December 2009

Background

Wolbachia(wBm) is an obligate endosymbiotic bacterium ofBrugia malayi, a parasitic filarial nematode of humans and one of the causative agents of lymphatic filariasis. There is a pressing need for new drugs against filarial parasites, such asB. malayi. AswBm is required forB. malayidevelopment and fertility, targetingwBm is a promising approach. However, the lifecycle of neitherB. malayinorwBm can be maintainedin vitro. To facilitate selection of potential drug targets we computationally ranked thewBm genome based on confidence that a particular gene is essential for the survival of the bacterium.

Results

wBm protein sequences were aligned using BLAST to the Database of Essential Genes (DEG) version 5.2, a collection of 5,260 experimentally identified essential genes in 15 bacterial strains. A confidence score, the Multiple Hit Score (MHS), was developed to predict eachwBm gene's essentiality based on the top alignments to essential genes in each bacterial strain. This method was validated using a jackknife methodology to test the ability to recover known essential genes in a control genome. A second estimation of essentiality, the Gene Conservation Score (GCS), was calculated on the basis of phyletic conservation of genes acrossWolbachia'sparent orderRickettsiales. Clusters of orthologous genes were predicted within the 27 currently available complete genomes. Druggability ofwBm proteins was predicted by alignment to a database of protein targets of known compounds.

Conclusion

RankingwBm genes by either MHS or GCS predicts and prioritizes potentially essential genes. Comparison of the MHS to GCS produces quadrants representing four types of predictions: those with high confidence of essentiality by both methods (245 genes), those highly conserved acrossRickettsiales(299 genes), those similar to distant essential genes (8 genes), and those with low confidence of essentiality (253 genes). These data facilitate selection ofwBm genes for entry into drug design pipelines.
Comparison of the sensitivity of culture, PCR and quantitative real-time PCR for the detection of Pseudomonas aeruginosain sputum of cystic fibrosis patients
Authors: Deschaght Pieter , De Baere Thierry , Van Simaey Leen , Van daele Sabine , De Baets Frans , De Vos Daniel , Pirnay Jean-Paul , Vaneechoutte Mario
Published: December 2009

Background

Pseudomonas aeruginosa is the major pathogen involved in the decline of lung function in cystic fibrosis (CF) patients. Early aggressive antibiotic therapy has been shown to be effective in preventing chronic colonization. Therefore, early detection is important and sensitive detection methods are warranted. In this study, we used a dilution series of P. aeruginosa positive sputa, diluted in a pool of P. aeruginosa negative sputa, all from CF patients - to mimick as closely as possible the sputa sent to routine laboratories - to compare the sensitivity of three culture techniques versus that of two conventional PCR formats and four real-time PCR formats, each targeting the P. aeruginosa oprL gene. In addition, we compared five DNA-extraction protocols.

Results

In our hands, all three culture methods and the bioMérieux easyMAG Nuclisens protocol Generic 2.0.1, preceded by proteinase K pretreatment and followed by any of the 3 real-time PCR formats with probes were most sensitive and able to detect P. aeruginosa up to 50 cfu/ml, i.e. the theoretical minimum of one cell per PCR mixture, when taking into account the volumes used in this study of sample for DNA-extraction, of DNA-elution and of DNA-extract in the PCR mixture.

Conclusion

In this study, no difference in sensitivity could be found for the detection of P. aeruginosa from sputum between microbiological culture and optimized DNA-extraction and real-time PCR. The results also indicate the importance of the optimization of the DNA-extraction protocol and the PCR format.
Subcutaneous administration of TC007 reduces disease severity in an animal model of SMA
Authors: Mattis Virginia B , Fosso Marina Y , Chang Cheng-Wei , Lorson Christian L
Published: December 2009

Background

Spinal Muscular Atrophy (SMA) is the leading genetic cause of infantile death. It is caused by the loss of functional Survival Motor Neuron 1 (SMN1). There is a nearly identical copy gene, SMN2, but it is unable to rescue from disease due to an alternative splicing event that excises a necessary exon (exon 7) from the majority of SMN2-derived transcripts. While SMNΔ7 protein has severely reduced functionality, the exon 7 sequences may not be specifically required for all activities. Therefore, aminoglycoside antibiotics previously shown to suppress stop codon recognition and promote translation read-through have been examined to increase the length of the SMNΔ7 C-terminus.

Results

Here we demonstrate that subcutaneous-administration of a read-through inducing compound (TC007) to an intermediate SMA model (Smn-/-; SMN2+/+; SMNΔ7) had beneficial effects on muscle fiber size and gross motor function.

Conclusion

Delivery of the read-through inducing compound TC007 reduces the disease-associated phenotype in SMA mice, however, does not significantly extend survival.
Far from easy and accurate - detection of metabolic syndrome by general practitioners
Authors: Helminen Eeva-Eerika , Mäntyselkä Pekka , Nykänen Irma , Kumpusalo Esko
Published: December 2009

Background

Metabolic syndrome (MetS) is a major public health challenge. General practitioners (GPs) could play a key role in its recognition. However, it often remains undiagnosed in primary care. This study assesses how well GPs and patients recognise MetS among patients with coronary heart disease or at least one of its risk factors.

Methods

Twenty-six health centres around Finland were randomly selected for the purpose of identifying, over a two-week period in April 2005, patients meeting the inclusion criteria of coronary heart disease or one of its risk factors. GPs and identified patients (n = 1880) were asked to complete surveys that included a question about the patient's MetS status. A trained nurse conducted health checks (n = 1180) of the identified patients, utilising criteria of MetS modified from the National Cholesterol Program. Data from the GPs' survey were compared with those from the health check to establish the extent of congruence of identification of MetS.

Results

Almost half (49.4%) of the patients met the criteria of MetS as established by objective measures. However, in the GPs' survey responses, only 28.5% of the patients were identified as having MetS. Additionally, these groups of MetS patients were not congruent. The sensitivity of the GPs' diagnosis of MetS was 0.31 with a specificity of 0.73. Only 7.1% of the study patients stated that they were suffering from MetS.

Conclusion

Detection of MetS is inaccurate among GPs in Finland. Most patients were not aware of having MetS. The practical relevance of MetS in primary care should be reconsidered.
Polymorphism in the oxytocin promoter region in patients with lactase non-persistence is not related to symptoms
Authors: Truedsson Mikael , Carlson Joyce , Simrén Magnus , Ohlsson Bodil
Published: December 2009

Background

Oxytocin and the oxytocin receptor have been demonstrated in the gastrointestinal (GI) tract and have been shown to exert physiological effects on gut motility. The role for oxytocin in the pathophysiology of GI complaints is unknown. The aim of this study was to examine genetic variations or polymorphism of oxytocin (OXT) and its receptor (OXTR) genes in patients with GI complaints without visible organic abnormalities.

Methods

Genetic variants in the OXT promoter region, and in the OXTR gene in DNA samples from 131 rigorously evaluated patients with Irritable Bowel Syndrome (IBS), 408 homozygous subjects referred for lactase (LCT-13910 C>T, rs4988235) genotyping, and 299 asymptomatic blood donors were compared. One polymorphism related to the OXT gene (rs6133010 A>G) and 4 related to the OXTR gene (rs1465386 G>T, rs3806675 G>A, rs968389 A>G, rs1042778 G>T) were selected for genotyping using Applied Biosystems 7900 HT allele discrimination assays.

Results

There were no statistically significant differences in the genotype or allele frequencies in any of the SNPs when IBS patients were compared to healthy controls. Among subjects referred for lactase genotyping, the rs6133010 A>G OXT promoter A/G genotype tended to be more common in the 154 non-persistent (27.3%) subjects than in the 254 lactase persistant (18.1%) subjects and in the healthy controls (19.4%) (p = 0.08). When direct comparing, the A/G genotype was less common in the OXT promoter region in controls (p = 0.09) and in subjects with lactase persistence (p = 0.03) compared to subjects with lactase non-persistence. When healthy controls were viewed according to their own LCT-13910 genotypes, the C/C lactase non-persistent controls had a higher frequency for the OXT promoter A/G genotype than LCT-13910 T/T lactase persistent controls (41.2% vs 13.1%).

No significant differences in frequencies of the investigated OXTR SNPs were noted in this study.

Conclusion

The results suggest that polymorphism in the promoter region of the OXT gene is most common in subjects with lactase non-persistence. This polymorphism may not be related to GI symptoms, as it is related to lactase non-persistence also in healthy controls.
A comparison of urinary tract pathology and morbidity in adult populations from endemic and non-endemic zones for urinary schistosomiasis on Unguja Island, Zanzibar
Authors: Lyons Beatrice , Stothard Russel , Rollinson David , Khamis Simba , Simai Khamis A , Hunter Paul R
Published: December 2009

Background

Renal tract involvement is implicated in both early and late schistosomiasis leading to increased disease burden. Despite there being good estimates of disease burden due to renal tract disease secondary to schistosomiasis at the global level, it is often difficult to translate these estimates into local communities. The aim of this study was to assess the burden of urinary tract pathology and morbidity due to schistosomiasis in Zanzibar and identify reliable clinical predictors of schistosomiasis associated renal disease.

Methods

A cross-sectional comparison of Ungujan men and women living within either high or low endemic areas for urinary schistosomiasis was conducted. Using urine analysis with reagent strips, parasitological egg counts, portable ultrasonography and a qualitative case-history questionnaire. Data analysis used single and multiple predictor variable logistic regression.

Results

One hundred and sixty people were examined in the high endemic area (63% women and 37% men), and 101 people in the low endemic area (61% women and 39% men). In the high endemic area, egg-patent schistosomiasis and urinary tract pathology were much more common (p = 1 × 10-3, 8 × 10-6, respectively) in comparison with the low endemic area. Self-reported frothy urine, self-reported haematuria, dysuria and urgency to urinate were associated with urinary tract pathology (p = 1.8 × 10-2, p = 1.1 × 10-4, p = 1.3 × 10-6, p = 1.1 × 10-7, respectively) as assessed by ultrasonography. In a multi-variable logistic regression model, self-reporting of schistosomiasis in the past year, self-reporting of urgency to urinate and having an egg-positive urine sample were all independently associated with detectable urinary tract abnormality, consistent with schistosomiasis-specific disease. Having two or more of these features was moderately sensitive (70%) as a predictor for urinary tract abnormality with high specificity (92%).

Conclusion

Having two out of urgency to urinate, self reporting of previous infections and detection of eggs in the urine were good proxy predictors of urinary tract abnormality as detected by ultrasound.
Evaluation of alternative respiratory syndromes for specific syndromic surveillance of influenza and respiratory syncytial virus: a time series analysis
Authors: Schindeler Suzanne K , Muscatello David J , Ferson Mark J , Rogers Kris D , Grant Paul , Churches Tim
Published: December 2009

Background

Syndromic surveillance is increasingly being evaluated for its potential for early warning of increased disease activity in the population. However, interpretation is hampered by the difficulty of attributing a causative pathogen. We described the temporal relationship between laboratory counts of influenza and respiratory syncytial virus (RSV) detection and alternative groupings of Emergency Department (ED) respiratory diagnoses.

Methods

ED and laboratory data were obtained for the south-eastern area of Sydney, NSW for the period 1 June 2001 - 1 December 2006. Counts of ED visits and laboratory confirmed positive RSV and influenza cases were aggregated by week. Semi-parametric generalized additive models (GAM) were used to determine the association between the incidence of RSV and influenza and the incidence of respiratory syndrome ED presentations while controlling for temporal confounders.

Results

For every additional RSV laboratory count, ED diagnoses of bronchiolitis increased by 3.1% (95%CI: 2.7%-3.5%) in the same week. For every additional influenza laboratory count, ED diagnoses of influenza-like illness increased by 4.7% (95%CI: 4.2%-5.2%) one week earlier.

Conclusion

In this study, large increases in ED diagnoses of bronchiolitis and influenza-like illness were independent and proxy indicators for RSV and influenza activity, respectively.
Further analysis of previously implicated linkage regions for Alzheimer's disease in affected relative pairs
Authors: Blom Elin S , Giedraitis Vilmantas , Arepalli Sampath , Hamshere Marian L , Adighibe Omanma , Goate Alison , Williams Julie , Lannfelt Lars , Hardy John , Vrièze Fabienne Wavrant-De , Glaser Anna
Published: December 2009

Background

Genome-wide linkage studies for Alzheimer's disease have implicated several chromosomal regions as potential loci for susceptibility genes.

Methods

In the present study, we have combined a selection of affected relative pairs (ARPs) from the UK and the USA included in a previous linkage study by Myers et al. (Am J Med Genet, 2002), with ARPs from Sweden and Washington University. In this total sample collection of 397 ARPs, we have analyzed linkage to chromosomes 1, 9, 10, 12, 19 and 21, implicated in the previous scan.

Results

The analysis revealed that linkage to chromosome 19q13 close to the APOE locus increased considerably as compared to the earlier scan. However, linkage to chromosome 10q21, which provided the strongest linkage in the previous scan could not be detected.

Conclusion

The present investigation provides yet further evidence that 19q13 is the only chromosomal region consistently linked to Alzheimer's disease.
Expression of indoleamine 2,3-dioxygenase in nasopharyngeal carcinoma impairs the cytolytic function of peripheral blood lymphocytes
Authors: Liu Peng , Xie Bai-Lu , Cai Shao-Hui , He Yun-Wen , Zhang Ge , Yi Yan-Mei , Du Jun
Published: December 2009

Background

Tumor-specific cytotoxic T cells and infiltrating lymphocytes are frequently found in tumor tissues in patients with nasopharyngeal carcinoma (NPC). Most patients with NPC, however, especially those with advanced stages, have a poor clinical prognosis despite conventional immunotherapy. The aim of this work was to examine the effect of indoleamine 2,3-dioxygenase (IDO), an immunosuppressive enzyme, on the lymphocyte function in NPC.

Methods

The NPC cell line CNE2 was treated by interferon-γ (IFNγ) and the levels of IDO expression was analyzed by Western blotting and reverse phase high-performance liquid chromatography (HPLC). Lymphocytes from health human exposed to the milieu created by IDO-positive CNE2 cells and the lymphocyte cytotoxicity to target tumor cells was analyzed by standard lactate dehydrogenase (LDH) release assay. Additionally, expression of IDO was determined by Immunohistochemical assay in the tumor tissues form clinically evaluated NPC.

Results

IDO expression was acutely induced in the NPC cell line CNE2 by low dose interferon-γ (IFNγ) or by co-incubation with activated lymphocytes. Exposure to the milieu created by IDO-positive CNE2 cells did not promote lymphocyte death, but lymphocyte cytotoxicity against target tumor cells was impaired. The suppression of lymphocyte cytotoxic function was fully restored when the conditioned medium was replaced by fresh medium for 24 h. In additionally, the IDO-positive cells were found scattered in the tumor tissues from patients with NPC.

Conclusion

Altogether, these findings suggest that IDO-mediated immunosuppression may be involved in the tumor immune evasion, and that blocking IDO activity in tumor cells may help to re-establish an effective anti-tumor T cell response in NPC.
Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes
Authors: Tardif Ginette , Hum David , Pelletier Jean-Pierre , Duval Nicolas , Martel-Pelletier Johanne
Published: December 2009

Background

MMP-13 and IGFBP-5 are important factors involved in osteoarthritis (OA). We investigated whether two highly predicted microRNAs (miRNAs), miR-140 and miR-27a, regulate these two genes in human OA chondrocytes.

Methods

Gene expression was determined by real-time PCR. The effect of each miRNA on IGFBP-5 and MMP-13 expression/production was evaluated by transiently transfecting their precursors (pre-miRNAs) and inhibitors (anti-miRNAs) into human OA chondrocytes. Modulation of IGFBP-5, miR-140 and miR-27a expression was determined upon treatment of OA chondrocytes with cytokines and growth factors.

Results

IGFBP-5 was expressed in human chondrocytes with its level significantly lower (p < 0.04) in OA. Five computational algorithms identified miR-140 and miR-27a as possible regulators of MMP-13 and IGFBP-5 expression. Data showed that both miRNAs were expressed in chondrocytes. There was a significant reduction (77%, p < 0.01) in miR-140 expression in OA compared to the normal chondrocytes, whereas miR-27a expression was only slightly decreased (23%). Transfection with pre-miR-140 significantly decreased (p = 0.0002) and with anti-miR-140 significantly increased (p = 0.05) IGFBP-5 expression at 24 hours, while pre-miR-27a did not affect either MMP-13 or IGFBP-5. Treatment with anti-miR-27a, but not with anti-miR-140, significantly increased the expression of both MMP-13 (p < 0.05) and IGFBP-5 (p < 0.01) after 72 hours of incubation. MMP-13 and IGFBP-5 protein production followed the same pattern as their expression profile. These data suggest that IGFBP-5 is a direct target of miR-140, whereas miR-27a down-regulates, likely indirectly, both MMP-13 and IGFBP-5.

Conclusion

This study is the first to show the regulation of these miRNAs in human OA chondrocytes. Their effect on two genes involved in OA pathophysiology adds another level of complexity to gene regulation, which could open up novel avenues in OA therapeutic strategies.
Oral administration of PPC enhances antigen-specific CD8+ T cell responses while reducing IgE levels in sensitized mice
Authors: Burrows Mike , Assundani Deepak , Celis Esteban , Tufaro Frank , Tanaka Akiko , Bradley W Guy
Published: December 2009

Background

For almost 2000 years it has been recognized that aqueous extracts from pine cones possess medicinal properties beneficial for the treatment of a broad variety of diseases and conditions. In this report, the ability of an orally administered poly phenylpropanoid-polysaccharide rich extract of pine cones (PPC) to suppress the generation of IgE and to significantly enhance antigen-specific cellular responses to a variety of vaccines was tested.

Methods

A variety of vaccine protocols were utilized to determine the affects of orally administered PPC on the Th1/Th2 cytokine balance, the production of IgE antibodies, and the generation of antigen-specific cytotoxic T cells. The effect of PPC on the Th1/Th2 balance in aged mice was also investigated.

Results

Oral delivery of PPC was found to significantly suppress serum IgE levels in naïve mice and in mice sensitized to ovalbumin. PPC was also found to enhance the generation of antigen-specific CD8+T cells in mice immunized with DNA, dendritic cell, and soluble protein vaccines. The suppression of IgE was associated with reduction of IL-4 secretion and the enhanced production of IL-12 and IFNγby antigen-stimulated splenocytes from PPC treated mice. PPC also suppressed the Th2 response and enhanced the Th1 response of splenocytes from aged mice.

Conclusion

Oral delivery of PPC enhances the generation of an antigen-specific CD8+T cell responses induced by soluble protein, DNA, and dendritic cell vaccines while at the same time suppressing the generation of a Th2 dominant IgE response. This effect on the Th1/Th2 balance was also observed in aged mice.
Trends in health services utilization, medication use, and health conditions among older adults: a 2-year retrospective chart review in a primary care practice
Authors: Vegda Ketan , Nie Jason X , Wang Li , Tracy C Shawn , Moineddin Rahim , Upshur Ross EG
Published: December 2009

Background

Population aging poses significant challenges to primary care providers and healthcare policy makers. Primary care reform can alleviate the pressures, but these initiatives require clinical benchmarks and evidence regarding utilization patterns. The objectives of this study is to measure older patients' use of health services, number of health conditions, and use of medications at the level of a primary care practice, and to investigate age- and gender-related utilization trends.

Methods

A cross-sectional chart audit over a 2-year study period was conducted in the academic family practice clinic of Sunnybrook Health Sciences Centre in Toronto, Ontario, Canada. All patients 65 years and older (n = 2450) were included. Main outcome measures included the number of family physician visits, specialist visits, emergency room visits, surgical admissions, diagnostic test days, inpatient hospital admissions, health conditions, and medications.

Results

Older patients (80-84 and 85+ age-group) had significantly more family physician visits (average of 4.4 visits per person per year), emergency room visits (average of 0.22 ER visits per year per patient), diagnostic days (average of 5.1 test days per person per year), health conditions (average of 7.7 per patient), and medications average of 8.2 medications per person). Gender differences were also observed: females had significantly more family physician visits and number of medications, while men had more specialist visits, emergency room visits, and surgical admissions. There were no gender differences for inpatient hospital admissions and number of health conditions. With the exception of the 85+ age group, we found greater intra-group variability with advancing age.

Conclusion

The data present a map of greater interaction with and dependency on the health care system with advancing age. The magnitudes are substantial and indicate high demands on patients and families, on professional health care providers, and on the health care system itself. There is the need to create and evaluate innovative models of care of multiple chronic conditions in the late life course.
Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa
Authors: Shafi Gowhar , Munshi Anjana , Hasan Tarique N , Alshatwi Ali A , Jyothy A , Lei David KY
Published: December 2009

Background

Cancer remains one of the most dreaded diseases causing an astonishingly high death rate, second only to cardiac arrest. The fact that conventional and newly emerging treatment procedures like chemotherapy, catalytic therapy, photodynamic therapy and radiotherapy have not succeeded in reverting the outcome of the disease to any drastic extent, has made researchers investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. This study progresses in the direction of identifying component(s) from Nigella sativa with anti cancer acitivity. In the present study we investigated the efficacy of Organic extracts of Nigella sativa seed powder for its clonogenic inhibition and induction of apoptosis in HeLa cancer cell.

Results

Methanolic, n-Hexane and chloroform extracts of Nigella sativa seedz effectively killed HeLa cells. The IC50 values of methanolic, n-hexane, and chloroform extracts of Nigella sativa were 2.28 μg/ml, 2.20 μg/ml and 0.41 ng/ml, respectively. All three extracts induced apoptosis in HeLa cells. Apoptosis was confirmed by DNA fragmentation, western blot and terminal transferase-mediated dUTP-digoxigenin-end labeling (TUNEL) assay.

Conclusion

Western Blot and TUNEL results suggested that Nigella sativa seed extracts regulated the expression of pro- and anti- apoptotic genes, indicating its possible development as a potential therapeutic agent for cervical cancer upon further investigation.
Dual effects of TGF-β on ERα-mediated estrogenic transcriptional activity in breast cancer
Authors: Ren Yongsheng , Wu Liyu , Frost Andra R , Grizzle William , Cao Xu , Wan Mei
Published: December 2009

Background

TGF-β resistance often develops in breast cancer cells that in turn overproduce this cytokine to create a local immunosuppressive environment that fosters tumor growth and exacerbates the invasive and metastatic behavior of the tumor cells themselves. Smads-mediated cross-talk with the estrogen receptor has been implied to play an important role in development and/or progression of breast cancer. We investigated how TGF-β regulates ERα-induced gene transcription and potential mechanisms of frequent TGF-β resistance in breast cancer.

Methods

Effect of TGF-β on ERα-mediated gene transcription was investigated in breast cancer cell lines using transient transfection, real-time PCR, sequential DNA precipitation, and small interfering RNA assays. The expression of Smads on both human breast cancer cell lines and ERα-positive human breast cancer tissue was evaluated by immunofluorescence and immunohistochemical assays.

Results

A complex of Smad3/4 mediates TGF-β inhibition of ERα-mediated estrogenic activity of gene transcription in breast cancer cells, and Smad4 is essential and sufficient for such repression. Either overexpression of Smad3 or inhibition of Smad4 leads to the "switch" of TGF-β from a repressor to an activator. Down-regulation and abnormal cellular distribution of Smad4 were associated with some ERα-positive infiltrating human breast carcinoma. There appears a dynamic change of Smad4 expression from benign breast ductal tissue to infiltrating ductal carcinoma.

Conclusion

These results suggest that aberrant expression of Smad4 or disruption of Smad4 activity lead to the loss of TGF-β suppression of ERα transactivity in breast cancer cells.
GLI1 genotypes do not predict basal cell carcinoma risk: a case control study
Authors: Watson Andrea , Kent Paul , Alam Murad , Paller Amy S , Umbach David M , Yoon Joon Won , Iannaccone Philip M , Walterhouse David O
Published: December 2009

Background

Susceptibility to basal cell carcinoma results from complex interactions between ultraviolet radiation exposure and genetic factors. The GLI1 oncogene is believed to play a role in the genesis of these tumors. We determined whether GLI1 polymorphisms were risk factors for developing basal cell carcinoma, either alone or in combination with patterns of past sun exposure, and whether there were functional differences among different GLI1 haplotypes.

Results

GLI1 genotypes at c.2798 and c.3298 from 201 basal cell carcinoma patients were compared to 201 age and sex-matched controls. Neither genotype nor haplotype frequencies differed between cases and controls. However, the odds of developing basal cell carcinoma on the trunk compared to the head/neck appeared somewhat lower with carriers of the c.3298GC than the CC genotype. There was no evidence for interactions between skin type, childhood sunburning, average adult sun exposure, adult sunbathing, or intermittency of sun exposure and GLI1 haplotype. Additionally, we found no significant differences in transcription activation or cell transforming ability among the four GLI1 haplotypes.

Conclusion

These results suggest that different GLI1 genotypes alone or in combination with past sun exposure patterns as assessed in this study do not affect basal cell carcinoma risk.
Analysis of recently identified dyslipidemia alleles reveals two loci that contribute to risk for carotid artery disease
Authors: Ronald James , Rajagopalan Ramakrishnan , Ranchalis Jane E , Marshall Julieann K , Hatsukami Thomas S , Heagerty Patrick J , Jarvik Gail P
Published: December 2009

Background

Genome-wide association studies have identified numerous single nucleotide polymorphisms (SNPs) affecting high density lipoprotein (HDL) or low density lipoprotein (LDL) cholesterol levels; these SNPs may contribute to the genetic basis of vascular diseases.

Results

We assessed the impact of 34 SNPs at 23 loci on dyslipidemia, key lipid sub-phenotypes, and severe carotid artery disease (CAAD) in a case-control cohort. The effects of these SNPs on HDL and LDL were consistent with those previously reported, and we provide unbiased estimates of the percent variance in HDL (3.9%) and LDL (3.3%) explained by genetic risk scores. We assessed the effects of these SNPs on HDL subfractions, apolipoprotein A-1, LDL buoyancy, apolipoprotein B, and lipoprotein (a) and found that rs646776 predicts apolipoprotein B level while rs2075650 predicts LDL buoyancy. Finally, we tested the role of these SNPs in conferring risk for ultrasonographically documented CAAD stenosis status. We found that two loci, chromosome 1p13.3 near CELSR2 and PSRC1 which contains rs646776, and 19q13.2 near TOMM40 and APOE which contains rs2075650, harbor risk alleles for CAAD.

Conclusion

Our analysis of 34 SNPs contributing to dyslipidemia at 23 loci suggests that genetic variation in the 1p13.3 region may increase risk of CAAD by increasing LDL particle number, whereas variation in the 19q13.2 region may increase CAAD risk by promoting formation of smaller, denser LDL particles.
Isolation and characterization of a strain of Lichtheimia corymbifera (ex Absidia corymbifera) from a case of bovine abortion
Authors: Piancastelli Chiara , Ghidini Francesca , Donofrio Gaetano , Jottini Stefano , Taddei Simone , Cavirani Sandro , Cabassi Clotilde S
Published: December 2009

Background

Lichtheimia corymbifera (previously Absidia corymbifera) is a filamentous zygomycetes belonging to the order Mucorales and to the family Lichtheimiaceae. Members of genus Lichtheimia spp. are cosmopolitan and ubiquitous in nature. Lichtheimia corymbifera is a recognized agent of diseases in man and animals. In cattle it causes abortion and mastitis. Three cases of bovine abortion occurred in a herd located in the Po Valley. Serological examinations were performed on fetal and mother's blood. One of the aborted fetus was referred to our laboratory. The paper describes the isolation and characterization of Lichtheimia corymbifera from a bovine aborted fetus.

Methods

Serological examinations were performed on fetal and mother's blood. Lesions on fetal tissues and placenta leaded the diagnostic suspect towards a mycotic aetiology. Tissues were then put in culture, and at the same time an histological examination was performed, together with bacteriological and virological tests. The isolate from placenta and fetal tissues was identified and characterized by PCR and RFLP, using the ITS region as a target sequence and AclI restriction site within the amplicon to distinguish Lichtheimia corymbifera among the other fungi.

Results

Serological, bacteriological and virological tests gave aspecific results. Histological examination evidenced numerous PAS positive hyphae within the necrotic cotiledons and numerous fungal nonseptate hyphae to the GMS stain. Colonies with typical morphological features of fungi grew up on Sabouraud agar from fetal skin and placenta. On the developed colonies the microscopic examination has shown a large number of nonseptate hyphae and sporangia consistent with Mucorales. PCR and RFLP allowed the identification of the isolate as Lichtheimia corymbifera.

Conclusion

The present report describes the isolation and the molecular characterisation of a fungal isolate from bovine aborted fetus and placenta. The diagnostic protocol allowed to identify and characterise the strain. This is the first isolation in Italy of Lichtheimia corymbifera in a bovine aborted fetus.
Proteomic analysis of blastema formation in regenerating axolotl limbs
Authors: Rao Nandini , Jhamb Deepali , Milner Derek J , Li Bingbing , Song Fengyu , Wang Mu , Voss S Randal , Palakal Mathew , King Michael W , Saranjami Behnaz , Nye Holly LD , Cameron Jo Ann , Stocum David L
Published: December 2009

Background

Following amputation, urodele salamander limbs reprogram somatic cells to form a blastema that self-organizes into the missing limb parts to restore the structure and function of the limb. To help understand the molecular basis of blastema formation, we used quantitative label-free liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS)-based methods to analyze changes in the proteome that occurred 1, 4 and 7 days post amputation (dpa) through the mid-tibia/fibula of axolotl hind limbs.

Results

We identified 309 unique proteins with significant fold change relative to controls (0 dpa), representing 10 biological process categories: (1) signaling, (2) Ca2+ binding and translocation, (3) transcription, (4) translation, (5) cytoskeleton, (6) extracellular matrix (ECM), (7) metabolism, (8) cell protection, (9) degradation, and (10) cell cycle. In all, 43 proteins exhibited exceptionally high fold changes. Of these, the ecotropic viral integrative factor 5 (EVI5), a cell cycle-related oncoprotein that prevents cells from entering the mitotic phase of the cell cycle prematurely, was of special interest because its fold change was exceptionally high throughout blastema formation.

Conclusion

Our data were consistent with previous studies indicating the importance of inositol triphosphate and Ca2+ signaling in initiating the ECM and cytoskeletal remodeling characteristic of histolysis and cell dedifferentiation. In addition, the data suggested that blastema formation requires several mechanisms to avoid apoptosis, including reduced metabolism, differential regulation of proapoptotic and antiapoptotic proteins, and initiation of an unfolded protein response (UPR). Since there is virtually no mitosis during blastema formation, we propose that high levels of EVI5 function to arrest dedifferentiated cells somewhere in the G1/S/G2 phases of the cell cycle until they have accumulated under the wound epidermis and enter mitosis in response to neural and epidermal factors. Our findings indicate the general value of quantitative proteomic analysis in understanding the regeneration of complex structures.
The use of body weight support on ground level: an alternative strategy for gait training of individuals with stroke
Authors: Sousa Catarina O , Barela José A , Prado-Medeiros Christiane L , Salvini Tania F , Barela Ana MF
Published: December 2009

Background

Body weight support (BWS) systems on treadmill have been proposed as a strategy for gait training of subjects with stroke. Considering that ground level is the most common locomotion surface and that there is little information about individuals with stroke walking with BWS on ground level, it is important to investigate the use of BWS on ground level in these individuals as a possible alternative strategy for gait training.

Methods

Thirteen individuals with chronic stroke (four women and nine men; mean age 54.46 years) were videotaped walking on ground level in three experimental conditions: with no harness, with harness bearing full body weight, and with harness bearing 30% of full body weight. Measurements were recorded for mean walking speed, cadence, stride length, stride speed, durations of initial and terminal double stance, single limb support, swing period, and range of motion of ankle, knee, and hip joints; and foot, shank, thigh, and trunk segments.

Results

The use of BWS system leads to changes in stride length and speed, but not in stance and swing period duration. Only the hip joint was influenced by the BWS system in the 30% BWS condition. Shank and thigh segments presented less range of motion in the 30% BWS condition than in the other conditions, and the trunk was held straighter in the 30% BWS condition than in the other conditions.

Conclusion

Individuals with stroke using BWS system on ground level walked slower and with shorter stride length than with no harness. BWS also led to reduction of hip, shank, and thigh range of motion. However, this system did not change walking temporal organization and body side asymmetry of individuals with stroke. On the other hand, the BWS system enabled individuals with chronic stroke to walk safely and without physical assistance. In interventions, the physical therapist can watch and correct gait pattern in patients' performance without the need to provide physical assistance.
Human γδ T cell Recognition of lipid A is predominately presented by CD1b or CD1c on dendritic cells
Authors: Cui Yongchun , Kang Lei , Cui Lianxian , He Wei
Published: December 2009

Background

The γδ T cells serve as early immune defense against certain encountered microbes. Only a few γδ T cell-recognized ligands from microbial antigens have been identified so far and the mechanisms by which γδ T cells recognize these ligands remain unknown. Here we explored the mechanism of interaction of human γδ T cells in peripheral blood with Lipid A (LA).

Results

First, resting γδ T cells (mainly Vδ2 T cells) displayed a strong proliferative response to LA-pulsed monocyte-derived dendritic cells (moDC) and LA-pulsed paraformaldehyde-fixed moDC, but not to free LA in a TCR γδ-dependent manner. Second, anti-CD1b or anti-CD1c antibodies could block proliferative response of resting γδ T cells to LA-loaded moDC. Besides, only LA-loaded CD1b/CD1c-transfected C1R lymphoblastoma cells (CD1b-/CD1c-C1R) were able to stimulate the proliferation of human γδ T cells. Third, the expressions of both Toll-like receptor (TLR)2 and TLR4 on surface of LA-activated γδ T cells were upregulated, whereas only anti-TLR4 antibody could partially block their response to LA; Finally LA-loaded moDCs induce γδ T cells to produce Th1 cytokines, such as IFN-γ.

Conclusion

Taken together, we found a novel mechanism that human γδ T cells recognize LA in a CD1b- or CD1c-restricted manner in first response against Gram-bacteria, while the interaction between TLR4 on γδ T cells and LA might strengthen the subsequent response of γδ T cells.

Reviewers

This article was reviewed by Hao Shen, Youwen He (nominated by Dr. Laurence C Eisenlohr), Dr. Michael Lenardo and Dr. Pushpa Pandiyan.
Crop and non-crop productivity in a traditional maize agroecosystem of the highland of Mexico
Authors: González-Amaro Rosa M , Martínez-Bernal Angélica , Basurto-Peña Francisco , Vibrans Heike
Published: December 2009

Background

In Mexico, the traditional maize cultivation system has resisted intensification attempts for many decades in some areas, even in some well-connected regions of the temperate highlands. We suggest that this is due to economics.

Methods

The total useful biomass of several fields in Nanacamilpa, Tlaxcala, are evaluated for productivity and costs.

Results

Maize grain production is low (1.5 t ha-1) and does not cover costs. However, maize stover demands a relatively high price. If it included, a profit is possible (about 110 US $ ha-1). We show that non-crop production (weeds for food and forage) potentially has a higher value than the crop. It is only partially used, as there are constraints on animal husbandry, but it diversifies production and plays a role as a back-up system in case of crop failure.

Conclusion

The diversified system described is economically rational under current conditions and labor costs. It is also stable, low-input and ecologically benign, and should be recognized as an important example of integrated agriculture, though some improvements could be investigated.
Mesothelioma of the testis and nephrotic syndrome: a case report
Authors: Bacchetta Justine , Ranchère Dominique , Dijoud Frédérique , Droz Jean-Pierre
Published: December 2009

Introduction

Paraneoplastic glomerulopathies are rare manifestations of neoplastic disease to be distinguished from iatrogenic renal damage. Solid tumors are preferentially associated with membranous nephropathy, whereas Hodgkin's lymphomas are associated with minimal change disease.

Case presentation

We report a 63-year-old Caucasian male diagnosed with a mesothelioma of the tunica vaginalis testis who, secondary to this, also presented with a nephrotic syndrome due to minimal change disease. In the present case, the paraneoplastic etiology of the nephrotic syndrome can be discussed on four unusual elements: minimal change lesions were found; the glomerulopathy was very sensitive to corticosteroids; the nephrotic syndrome occurred 11 months after the diagnosis of the primary malignancy, but concomitantly with the recurrence; and the nephrotic syndrome did not decrease with tumor control and did not recur when the mesothelioma escaped treatment. No other etiologies could nevertheless explain this phenomenon.

Conclusion

Paraneoplastic nephrotic syndrome is often associated with membranous nephropathy in patients with solid tumors, especially in patients with lung and gastrointestinal tract neoplasia. The management of these patients is associated with a symptomatic treatment such as sodium and water restriction, diuretics and ACE inhibitors and a prophylaxis of specific complications of nephrotic syndrome including thromboembolism, infections and lipid abnormalities. Treatment of neoplasia must be undertaken rapidly, treatments must be regularly analyzed and drugs binding to albumin may be used with precaution.
Meiotic segregation and interchromosomal effect in the sperm of a double translocation carrier: a case report
Authors: Juchniuk de Vozzi Maria S , Santos Silvio A , Pereira Ciro S , Cuzzi Juliana F , Laureano Lucimar AF , Franco Jr José G , Martelli Lucia
Published: December 2009

Background

Infertility is a natural mechanism of selection intended to prevent the delivery of a child with malformations or mental retardation. Male infertility is closely related to chromosomal abnormalities. This study was focused on the analysis of meiotic segregation involving a Robertsonian translocation, 45,XY,der(13;13) [56]/45,XY,der(13;14) [44] and the evaluation of possible interchromosomal effects.

Results

Hybridisation with LSI 13q14 and subtelomere 14q probes and WCP13 SpectrumGreen and WCP14 SpectrumOrange probes showed a high proportion of unbalanced gametes, corresponding to 71.2% of the spermatozoa. The disomic frequencies of the sexual chromosomes and chromosome 18 of the patient were higher (5.28% and 2.55%, respectively) than those of the control (0.6% and 0.59%, respectively).

Conclusion

Meiotic segregation studies in sperm are an important tool for genetic counselling of chromosomal aberrations, allowing for a prediction of the risks and consequent implications for the reproductive life. The patient with this rare translocation exhibited meiotic segregation fidelity, and a high rate of unbalanced gametes with disomic spermatozoa.
Spontaneous bowel perforation due to norovirus: a case report
Authors: Pawa Nikhil , Vanezis Andrew P , Tutton Matthew G
Published: December 2009

Norovirus is the leading cause of epidemic gastroenteritis worldwide but the disease is usually self-limiting and generally only causes serious health problems in the young, elderly and immunocompromised. The authors report a case of bowel perforation in an elderly Caucasian lady with confirmed infection with Norovirus genogroup II and no other presumptive cause. To the authors' knowledge this is the first such case of bowel perforation due to Norovirus. Viral gastroenteritis should be considered in the list of differentials when no obvious cause of bowel perforation can be identified to minimise morbidity and mortality.