bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative effects of RRR-alpha- and RRR-gamma-tocopherol on proliferation and apoptosis in human colon cancer cell lines
Authors: Campbell Sharon E , Stone William L , Lee Steven , Whaley Sarah , Yang Hongsong , Qui Min , Goforth Paige , Sherman Devin , McHaffie Derek , Krishnan Koyamangalath
Published: January 2006

Background

Mediterranean societies, with diets rich in vitamin E isoforms, have a lower risk for colon cancer than those of northern Europe and the Americas. Vitamin E rich diets may neutralize free radicals generated by fecal bacteria in the gut and prevent DNA damage, but signal transduction activities can occur independent of the antioxidant function. The term vitamin E represents eight structurally related compounds, each differing in their potency and mechanisms of chemoprevention. The RRR-γ-tocopherol isoform is found primarily in the US diet, while RRR-α-tocopherol is highest in the plasma.

Methods

The effectiveness of RRR-α- and RRR-γ-tocopherol at inhibiting cell growth and inducing apoptosis in colon cancer cell lines with varying molecular characteristics (SW480, HCT-15, HCT-116 and HT-29) and primary colon cells (CCD-112CoN, nontransformed normal phenotype) was studied. Colon cells were treated with and without RRR-α- or RRR-γ-tocopherol using varying tocopherol concentrations and time intervals. Cell proliferation and apoptosis were measured using the trypan blue assay, annexin V staining, DNA laddering and caspase activation.

Results

Treatment with RRR-γ-tocopherol resulted in significant cell death for all cancer cell lines tested, while RRR-α-tocopherol did not. Further, RRR-γ-tocopherol treatment showed no cytotoxicity to normal colon cells CCD-112CoN at the highest concentration and time point tested. RRR-γ-tocopherol treatment resulted in cleavage of PARP, caspase 3, 7, and 8, but not caspase 9. Differences in the percentage cell death and apoptosis were observed in different cell lines suggesting that molecular differences in these cell lines may influence the ability of RRR-γ-tocopherol to induce cell death.

Conclusion

This is the first study to demonstrate that multiple colon cancer cell lines containing varying genetic alterations will under go growth reduction and apoptosis in the presence of RRR-γ-tocopherol without damage to normal colon cells. The amount growth reduction was dependent upon the molecular signatures of the cell lines. Since RRR-γ-tocopherol is effective at inhibition of cell proliferation at both physiological and pharmacological concentrations dietary RRR-γ-tocopherol may be chemopreventive, while pharmacological concentrations of RRR-γ-tocopherol may aid chemotherapy without toxic effects to normal cells demonstrated by most chemotherapeutic agents.
Angiomotin and angiomotin like proteins, their expression and correlation with angiogenesis and clinical outcome in human breast cancer
Authors: Jiang Wen G , Watkins Gareth , Douglas-Jones Anthony , Holmgren Lars , Mansel Robert E
Published: January 2006

Backgound

Angiomotin is a newly discovered molecule that regulates the migration and tubule formation of endothelial cells. It therefore has been implicated in the control of angiogenesis under physiological and pathological conditions. This study examined the expression of angiomotin and its analogues, angiomotin-like 1 (L1) and -like 2 (L2) in breast tumour tissues, and analysed their correlation with angiogenesis and clinical outcomes.

Methods

Human breast tissues (normal n = 32 and tumours n = 120) were used. The levels of expression of angiomotin, L1 and L2 were determined using reverse transcription PCR. Microvessels were stained using antibodies against PECAM, von Willebrand factor (factor 8, or vWF) and VE-cadherin. The transcript levels of angiomotin and its analogues were assessed against the clinical and pathological background, including long term survival (120 months).

Results

Breast cancer tissues expressed significantly higher levels of angiomotin transcript, compared with normal mammary tissues (33.1 ± 11 in normal versus 86.5 ± 13.7 in tumour tissues, p = 0.003). Both L1 and L2 were seen at marginally higher levels in tumour than normal tissues but the difference was not statistically significant. Levels of angiomotin were at significantly higher levels in grade 2 and grade 3 tumours compared with grade 1 (p < 0.01 and p = 0.05 respectively). The levels of angiomotin in tumours from patients who had metastatic disease were also significantly higher than those patients who remained disease free (p = 0.03). Multivariate analysis indicated that angiomotin transcript was an independent prognostic factor (p = 0.031). No significant correlations were seen between angiomotin-L1 and L2 with the clinical outcome. Furthermore, high levels of angiomotin transcript were associated with shorter overall survival (p < 0.05). There was a high degree of correlation between levels of vW factor and that of angiomotin (p < 0.05), but not angiomotin-L1 and angiomotin-L2.

Conclusion

Angiomotin, a putative endothelial motility factor, is highly expressed in human breast tumour tissues and linked to angiogenesis. It links to the aggressive nature of breast tumours and the long term survival of the patients. These data point angiomotin as being a potential therapeutic target.
Prognosis of screen-detected breast cancers: results of a population based study
Authors: Cortesi Laura , Chiuri Vincenzo E , Ruscelli Silvia , Bellelli Valeria , Negri Rossella , Rashid Ivan , Cirilli Claudia , Fracca Antonella , Gallo Ennio , Federico Massimo
Published: January 2006

Background

The reduced mortality rate from breast carcinoma among women offered screening mammography is demonstrated after 15–20 years of follow-up. However, the assessment of 5-year overall and event-free survival could represent an earlier measure of the efficacy of mammography screening program (MSP).

Methods

All cases of breast cancer diagnosed in the Province of Modena between years 1996 and 2000 in women aged 50 to 69 years, were identified through the Modena Cancer Registry (MCR). Stage of disease and treatment information were obtained from clinical records. All the events occurring up to June 30, 2003 were retrieved by experienced monitors. Five-year overall and event-free survival were the principal end-points of the study.

Results

During a 5-year period, 587 primary breast cancers were detected by the MSP and 471 primary breast cancers were diagnosed out of the MSP. The screen-detected breast cancers were smaller, more likely node negative, with low histological grade, low proliferative activity and positive receptors status. Furthermore, the breast cancer diagnosed through the MSP more frequently received a conservative surgery. The 5-year survival rate was 94% in the screen-detected group, versus 84% in the other group (p = 0.0001). The rate of 5-year event-free survival was 89% and 75% for the MSP participants and not participants, respectively (p = 0.0001).

Conclusions

Our data confirm a favourable outcome of screen-detected breast cancers in terms of five-year overall and event-free survival, which reflect the good quality assurance parameters of the MSP. Finally, a cancer registry should be implemented in every area covered by screening programs.
Upregulation of meiosis-specific genes in lymphoma cell lines following genotoxic insult and induction of mitotic catastrophe
Authors: Kalejs Martins , Ivanov Andrey , Plakhins Gregory , Cragg Mark S , Emzinsh Dzintars , Illidge Timothy M , Erenpreisa Jekaterina
Published: January 2006

Background

We have previously reported that p53 mutated radioresistant lymphoma cell lines undergo mitotic catastrophe after irradiation, resulting in metaphase arrest and the generation of endopolyploid cells. A proportion of these endopolyploid cells then undergo a process of de-polyploidisation, stages of which are partially reminiscent of meiotic prophase. Furthermore, expression of meiosis-specific proteins of the cancer/testis antigens group of genes has previously been reported in tumours. We therefore investigated whether expression of meiosis-specific genes was associated with the polyploidy response in our tumour model.

Methods

Three lymphoma cell lines, Namalwa, WI-L2-NS and TK6, of varying p53 status were exposed to a single 10 Gy dose of gamma radiation and their responses assessed over an extended time course. DNA flow cytometry and mitotic counts were used to assess the kinetics and extent of polyploidisation and mitotic progression. Expression of meiotic genes was analysed using RT-PCR and western blotting. In addition, localisation of the meiotic cohesin REC8 and its relation to centromeres was analysed by immunofluorescence.

Results

The principal meiotic regulator MOS was found to be significantly post-transcriptionally up-regulated after irradiation in p53 mutated but not p53 wild-type lymphoma cells. The maximum expression of MOS coincided with the maximal fraction of metaphase arrested cells and was directly proportional to both the extent of the arrest and the number of endopolyploid cells that subsequently emerged. The meiotic cohesin REC8 was also found to be up-regulated after irradiation, linking sister chromatid centromeres in the metaphase-arrested and subsequent giant cells. Finally, RT-PCR revealed expression of the meiosis-prophase genes, DMC1, STAG3, SYCP3 and SYCP1.

Conclusion

We conclude that multiple meiotic genes are aberrantly activated during mitotic catastrophe in p53 mutated lymphoma cells after irradiation. Furthermore, we suggest that the coordinated expression of MOS and REC8 regulate the extent of arrested mitoses and polyploidy.
The carcinogenic potential of tacrolimus ointment beyond immune suppression: a hypothesis creating case report
Authors: Becker Jürgen C , Houben Roland , Vetter Claudia S , Bröcker Eva B
Published: January 2006

Background

Since tacrolimus ointment was approved by the U.S. Food and Drug Administration (FDA) as a promising treatment for atopic dermatitis, it has been approved in more than 30 additional countries, including numerous European Union member nations. Moreover, in the current clinical routine the use of this drug is no longer restricted to the approved indication, but has been extended to a wide variety of inflammatory skin diseases including some with the potential of malignant transformation. So far, the side-effects reported from the topical use of tacrolimus have been relatively minor (e.g. burning, pruritus, erythema). Recently, however, the FDA reviewed the safety of topical tacrolimus, which resulted in a warning that the use of calcineurin inhibitors may be associated with an increased risk of cancer.

Case presentation

Oral lichen planus (OLP) was diagnosed in a 56-year-old women in February 1999. After several ineffective local and systemic therapeutic measures an off-label treatment of this recalcitrant condition using Tacrolimus 0.1% ointment was initiated in May 2002. After a few weeks of treatment most of the lesions ameliorated, with the exception of the plaques on the sides of the tongue. Nevertheless, the patient became free of symptoms which, however, reoccurred once tacrolimus was weaned, as a consequence treatment was maintained. In April 2005, the plaques on the left side of the tongue appeared increasingly compact and a biopsy specimen confirmed the suspected diagnosis of an oral squamous cell carcinoma.

Conclusion

The suspected causal relationship between topical use of tacrolimus and the development of a squamous cell carcinoma prompted us to test the notion that the carcinogenicity of tacrolimus may go beyond mere immune suppression. To this end, tacrolimus has been shown to have an impact on cancer signalling pathways such as the MAPK and the p53 pathway. In the given case, we were able to demonstrate that these pathways had also been altered subsequent to tacrolimus therapy.
Refractile superficial retinal crystals and chronic retinal detachment: Case report
Authors: Habib Maged S , Byrne Sinead , McCarthy John H , Steel David HW
Published: January 2006

Background

Few previous reports have described the presence of retinal refractile opacities at the macular area in patients presenting with longstanding peripheral retinal detachment. The exact nature of these opacities is unknown.

Case presentation

Two patients were referred with an abnormal appearance of refractile opacities in the macular area noted during routine examination. Both were found to have longstanding peripheral retinal detachments. Subretinal fluid analysis of one patient revealed the presence of multiple birefringent crystals. We hypothesise that these crystals are the origin of the refractile macular opacities noted.

Conclusion

This report describes the rare presentation of asymptomatic peripheral retinal detachment by the detection of refractile macular opacities on routine examination. It highlights the importance of meticulous peripheral retinal examination in these cases. The article also describes the findings of the subretinal fluid analysis and discusses the possible hypothesis behind their appearance.
Variation in Hepatitis C services may lead to inequity of heath-care provision: a survey of the organisation and delivery of services in the United Kingdom
Authors: Parkes Julie , Roderick Paul , Bennett-Lloyd Bethan , Rosenberg William
Published: January 2006

Background

Chronic hepatitis C infection (CHC) is a major healthcare problem. Effective anti-viral therapy is available. To maximise population effectiveness, co-ordinated services for detection and management of patients with CHC are required. There is a need to determine patterns of healthcare delivery to plan improvements. A study was conducted to determine workload, configuration and care processes of current UK services available to manage patients with CHC.

Methods

A cross-sectional questionnaire survey of consultant members of British Association for the Study of the Liver (n = 53), Infectious Disease consultants (n = 43), and a 1 in 5 sample of Genito-Urinary Medicine (n = 48) and gastroenterologists (n = 200).

Results

Response rate was 70%. 40% of respondents provided a comprehensive service (included treatment and follow-up): speciality of clinical leads identified as Hepatology (37%); Gastroenterology (47%); and Infectious Disease (16%). The estimated number of patients managed by respondents was about 23,000 with an upward trend over the previous 3 years. There was variation between comprehensive service providers, including unit size, eligibility criteria for treatment, and drug regimes. Key barriers to quality of care identified were staffing capacity, funding of treatment and patient non-attendance. Most English strategic health authorities had at least one comprehensive service provider.

Conclusion

There was significant variation in all aspects of the patient pathway which may contribute to inequity of health care provision. Services need to be expanded to form geographical clinical networks, and properly resourced to ensure greater uptake and more equitable delivery of services if the future burden of chronic liver disease is to be reduced.
Impact of vaccine economic programs on physician referral of children to public vaccine clinics: a pre-post comparison
Authors: Zimmerman Richard K , Tabbarah Melissa , Janosky Janine E , Bardenheier Barbara , Troy Judith A , Jewell Ilene K , Yawn Barbara P
Published: January 2006

Background

The Vaccines for Children (VFC) Program is a major vaccine entitlement program with limited long-term evaluation. The objectives of this study are to evaluate the effect of VFC on physician reported referral of children to public health clinics and on doses administered in the public sector.

Methods

Minnesota and Pennsylvania primary care physicians (n = 164), completed surveys before (e.g., 1993) and after (2003) VFC, rating their likelihood on a scale of 0 (very unlikely) to 10 (very likely) of referring a child to the health department for immunization.

Results

The percentage of respondents likely to refer was 60% for an uninsured child, 14% for a child with Medicaid, and 3% for a child with insurance that pays for immunization. Half (55%) of the physicians who did not participate in VFC were likely to refer a Medicaid-insured child, as compared with 6% of those who participated (P< 0.001). Physician likelihood to refer an uninsured child for vaccination, measured on a scale of 0 to 10 where 10 is very likely, decreased by a mean difference of 1.9 (P< 0.001) from pre- to post-VFC. The likelihood to refer a Medicaid-insured child decreased by a mean of 1.2 (P= 0.001).

Conclusion

Reported out-referral to public clinics decreased over time. In light of increasing immunizations rates, this suggests that more vaccines were being administered in private provider offices.
Quantitative ultrasound tissue characterization in shoulder and thigh muscles – a new approach
Authors: Nielsen Pernille Kofoed , Jensen Bente R , Darvann Tron , Jørgensen Kurt , Bakke Merete
Published: January 2006

Background

The echogenicity patterns of ultrasound scans contain information of tissue composition in muscles. The aim was: (1) to develop a quantitative ultrasound image analysis to characterize tissue composition in terms of intensity and structure of the ultrasound images, and (2) to use the method for characterization of ultrasound images of the supraspinatus muscle, and the vastus lateralis muscle.

Methods

Computerized texture analyses employing first-order and higher-order grey-scale statistics were developed to objectively characterize ultrasound images of m. supraspinatus and m. vastus lateralis from 9 healthy participants.

Results

The mean grey-scale intensity was higher in the vastus lateralis muscle (p < 0.05) than in the supraspinatus muscle (average value of middle measuring site 51.4 compared to 35.0). Furthermore, the number of spatially connected and homogeneous regions (blobs) was higher in the vastus lateralis (p < 0.05) than in the supraspinatus (average for m. vastus lateralis: 0.092 mm-2 and for m. supraspinatus: 0.016 mm-2).

Conclusion

The higher intensity and the higher number of blobs in the vastus lateralis muscle indicates that the thigh muscle contained more non-contractile components than the supraspinatus muscle, and that the muscle was coarser. The image analyses supplemented each other and gave a more complete description of the tissue composition in the muscle than the mean grey-scale value alone.
Sequence specific visual detection of LAMP reactions by addition of cationic polymers
Authors: Mori Yasuyoshi , Hirano Tsuyoshi , Notomi Tsugunori
Published: January 2006

Background

Development of a practical gene point-of-care testing device (g-POCT device) requires innovative detection methods for demonstrating the results of the gene amplification reaction without the use of expensive equipment. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using precipitation reaction by addition of cationic polymers to amplicons of Loop mediated isothermal Amplification (LAMP).

Results

Oligo DNA probes labeled with different fluorescent dyes were prepared for multiple nucleic acid templates, and the templates were amplified by the LAMP reactions under the existence of the probes. At completion of the LAMP reaction, an optimal amount of low molecular weight polyethylenimine (PEI) was added, resulting in the precipitation of the insoluble LAMP amplicon-PEI complex. The fluorescently labeled Oligo DNA probes hybridized to the LAMP product were incorporated into the precipitation, and the precipitate emitted fluorescence corresponding to the amplified nucleic acid templates. The color of emitted fluorescence can be detected easily by naked eye on a conventional UV illuminator.

Conclusion

The presence or absence of minute amount of nucleic acid templates could be detected in a simple manner through visual assessment for the color of the LAMP amplicon-PEI complex precipitate. We conclude that this detection method may facilitate development of small and simple g-POCT device.
The inflammatory and normal transcriptome of mouse bladder detrusor and mucosa
Authors: Saban Marcia R , Hellmich Helen L , Turner Mary , Nguyen Ngoc-Bich , Vadigepalli Rajanikanth , Dyer David W , Hurst Robert E , Centola Michael , Saban Ricardo
Published: January 2006

Background

An organ such as the bladder consists of complex, interacting set of tissues and cells. Inflammation has been implicated in every major disease of the bladder, including cancer, interstitial cystitis, and infection. However, scanty is the information about individual detrusor and urothelium transcriptomes in response to inflammation. Here, we used suppression subtractive hybridizations (SSH) to determine bladder tissue- and disease-specific genes and transcriptional regulatory elements (TRE)s. Unique TREs and genes were assembled into putative networks.

Results

It was found that the control bladder mucosa presented regulatory elements driving genes such as myosin light chain phosphatase and calponin 1 that influence the smooth muscle phenotype. In the control detrusor network the Pax-3 TRE was significantly over-represented. During development, the Pax-3 transcription factor (TF) maintains progenitor cells in an undifferentiated state whereas, during inflammation, Pax-3 was suppressed and genes involved in neuronal development (synapsin I) were up-regulated. Therefore, during inflammation, an increased maturation of neural progenitor cells in the muscle may underlie detrusor instability. NF-κB was specifically over-represented in the inflamed mucosa regulatory network. When the inflamed detrusor was compared to control, two major pathways were found, one encodingsynapsin I, a neuron-specific phosphoprotein, and the other an important apoptotic protein,siva. In response to LPS-induced inflammation, the liver X receptor was over-represented in both mucosa and detrusor regulatory networks confirming a role for this nuclear receptor in LPS-induced gene expression.

Conclusion

A new approach for understanding bladder muscle-urothelium interaction was developed by assembling SSH, real time PCR, and TRE analysis results into regulatory networks. Interestingly, some of the TREs and their downstream transcripts originally involved in organogenesis and oncogenesis were also activated during inflammation. The latter represents an additional link between inflammation and cancer. The regulatory networks represent key targets for development of novel drugs targeting bladder diseases.
Crystal structure of a DNA polymerase sliding clamp from a Gram-positive bacterium
Authors: Argiriadi Maria A , Goedken Eric R , Bruck Irina , O'Donnell Mike , Kuriyan John
Published: January 2006

Background

Sliding DNA clamps are processivity factors that are required for efficient DNA replication. DNA polymerases maintain proximity to nucleic acid templates by interacting with sliding clamps that encircle DNA and thereby link the polymerase enzyme to the DNA substrate. Although the structures of sliding clamps from Gram-negative bacteria (E. coli), eukaryotes, archaea, and T4-like bacteriophages are well-known, the structure of a sliding clamp from Gram-positive bacteria has not been reported previously.

Results

We have determined the crystal structure of the dimeric β subunit of the DNA polymerase III holoenzyme of Streptococcus pyogenes. The sliding clamp from this Gram-positive organism forms a ring-shaped dimeric assembly that is similar in overall structure to that of the sliding clamps from Gram-negative bacteria, bacteriophage T4, eukaryotes and archaea. The dimer has overall dimensions of ~90 Å × ~70 Å × ~25 Å with a central chamber that is large enough to accommodate duplex DNA. In comparison to the circular shape of other assemblies, the S. pyogenes clamp adopts a more elliptical structure.

Conclusion

The sequences of sliding clamps from S. pyogenes and E. coli are only 23% identical, making the generation of structural models for the S. pyogenes clamp difficult in the absence of direct experimental information. Our structure of the S. pyogenes β subunit completes the catalog of clamp structures from all the major sequence grouping of sliding clamps. The more elliptical rather than circular structure of the S. pyogenes clamp implies that the topological nature of encircling DNA, rather than a precise geometric shape, is the most conserved aspect for this family of proteins.
European active surveillance study of women taking HRT (EURAS-HRT): study protocol [NCT00214903]
Authors: Dinger Juergen C , Heinemann Lothar AJ
Published: January 2006

Background

The post marketing safety surveillance program for a drug containing a new chemical entity should assess both, the safety outcomes that relate specifically to the targeted population, as well as those that could potentially be related to special pharmacological characteristics of the drug. Active safety surveillance using valid epidemiological study designs has been proven to be a pertinent and reliable method to approach this endeavor.

Methods/design

The primary objective of the study is to compare incidence rates of serious adverse events in users of all types of newly prescribed oral HRT products. This active surveillance study will assess pertinent cardiovascular outcomes - in particular venous and arterial thromboembolism - and other serious adverse events (SAEs) in new HRT users over a period of several years. One product under surveillance is Angeliq®, which contains the novel progestagen drospirenone (DRSP) combined with estradiol. In addition, all other oral combined HRT products with a novel progestagen or estrogen that will be newly marketed during the study period will be studied. These new HRT products will be compared with established HRT products. The combined cohort will include at least 30,000 women recruited in several European countries. At least 90,000 years of observation are expected from the field work which started in early 2002 and will end around 2008. The participating women will complete a baseline survey using a self-administered questionnaire to describe the baseline risk. After 6 months, 12 months, and then on an annual basis, they will fill out a questionnaire in which they record complaints and events during the use of the prescribed HRTs. All adverse outcomes occurring during the observational period will be evaluated.

Discussion

A complete lifetime medical history, individually validated SAEs over time, and a low loss to follow-up rate are essential for a robust safety assessment. Therefore, the lifetime history of diseases and relevant medications will be documented. Reported SAEs will be validated and analyzed. A four level, multi-faceted follow-up process was established to ensure low loss to follow-up rates (e.g., 3–5% after three years of follow up). Multivariate methods will be used to adjust for confounding.
In vitro modulation of inflammatory cytokine and IgG levels by extracts of Perna canaliculus
Authors: Mani Sachin , Lawson John W
Published: January 2006

Background

Inflammation is a predominant characteristic of autoimmune diseases which is characterized by the increased expression of pro-inflammatory cytokines. Soon to be published work from our laboratory has shown that ingestion ofPerna canaliculusprevents the development of autoimmune diseases such as Systemic Lupus Erythematosus and rheumatoid arthritis in laboratory animals. The current paper attempts to illustrate how Perna can alleviate inflammation by modulating inflammatory cytokines, cyclooxygenase enzymes and Immunoglobulin-G (IgG) levels.

Methods

In the present study, hydrochloric acid [HCl] and Tween-20 were used to develop extracts of Perna. These extracts were assayed for protein content. Increasing concentrations of these extracts were then tested in cell culture for modulation of inflammatory cytokine, cyclooxygenase enzymes and IgG levels. Parallel tests were run using an available glycogen extract of Perna as a comparison to our in-house laboratory preparations.

Results

Tween-20 Perna extracts were found to be more stable and less toxic in cell culture than HCl digest of Perna. They also assayed higher in protein content that HCl extracts. Although both extracts inhibited IgG production in V2E9 hybridomas, Tween-20 extracts were more consistent in IgG suppression than HCl extracts. Overall Tween-20 extracts effectively decreased levels of TNF-α, IL-1, IL-2 and IL-6 as observed using cytokine bioassays. Twenty micrograms of Tween-20 Perna extracts induced such significant decreases in inflammatory cytokine production that when tested on sensitive cell lines, they very nearly abolished the decrease in viability induced by these cytokines. Tween-20 extracts effectively inhibited both COX-1 and COX-2 cyclooxygenase activity. As a comparison, the glycogen extract also demonstrated a similar though weaker effect on COX-1 and COX-2 enzymes. The active components of both extracts (Tween-20 and glycogen) were observed to possess molecular weights above 100 kDa. Although the anti-cytokine activity of the Tween-20 extract was destroyed by Proteinase-K treatment, the anti-COX-1 and anti-COX-2 activity of both the extracts were not sensitive to protease treatment.

Conclusion

We have successfully demonstrated modulation in the levels of inflammatory cytokines, cyclooxygenase enzymes and immunoglobulins by our in-house laboratory preparations ofPerna canaliculus, whereby suggesting an immunomodulatory role ofPerna canaliculusin regulating inflammation.
Inclusion of MUC1 (Ma695) in a panel of immunohistochemical markers is useful for distinguishing between endocervical and endometrial mucinous adenocarcinoma*
Authors: Khoury Thaer , Tan Dongfeng , Wang Jianmin , Intengan Marilyn , Yang Jun , Alrawi Sadir , Yan Peisha , Byrd James C
Published: January 2006

Background

Distinguishing endocervical adenocarcinoma (ECA) from endometrial mucinous adenocarcinoma (EMMA) is clinically significant in view of the differences in their management and prognosis. In this study, we used a panel of tumor markers to determine their ability to distinguish between primary endocervical adenocarcinoma and primary endometrial mucinous adenocarcinoma.

Methods

Immunohistochemistry using monoclonal antibodies to MUC1 (Ma695), p16, estrogen receptor (ER), progesterone receptor (PR), and vimentin, was performed to examine 32 cases, including 18 EMMAs and 14 ECAs. For MUC1, cases were scored based on the percentage of staining pattern, apical, apical and cytoplasmic (A/C), or negative. For p16, cases were scored based on the percentage of cells stained. For the rest of the antibodies, semiquantitative scoring system was carried out.

Results

For MUC1, majority of EMMA (14 of 18 cases, 78%) showed A/C staining, whereas only few ECA (2 of 14, 14%) were positive. The difference of MUC1 expression in the two groups of malignancy was statistically significant (p < 0.001). Staining for p16 was positive in 10 of 14 (71%) ECA and 4 of 18 (22%) EMMA. Estrogen receptor was positive in 3 of 14 (21%) ECA and 17 of 18 (94%) EMMA. Progesterone receptor was positive in 3 of 14 (21%) ECA and 16 of 18 (89%) EMMA. Vimentin was positive in 1 of 14 (7%) ECA, and 9 of 18 (50%) EMA, with median and range of 0 (0–6), and 1.5 (0–9) respectively.

Conclusion

A panel of immunohistochemical markers including MUC1, p16, ER, PR, and vimentin is recommended, when there is morphological and clinical doubt as to the primary site of endocervical or endometrial origin.
Real-time PCR complements immunohistochemistry in the determination of HER-2/neu status in breast cancer
Authors: Nistor Andreea , Watson Peter H , Pettigrew Norman , Tabiti Karim , Dawson Angelika , Myal Yvonne
Published: January 2006

Background

The clinical benefit of determining the status of HER-2/neu amplification in breast cancer patients is well accepted. Although immunohistochemistry (IHC) is the most frequently used method to assess the over-expression of HER-2 protein, fluorescent in-situ hybridization (FISH) is recognized as the "gold standard" for the determining of HER-2/neu status. The greatest discordance between the two methods occurs among breast tumors that receive an indeterminate IHC score of 2+. More recently, a real-time polymerase chain reaction (PCR) assay using the LightCycler® has been developed for quantifying HER-2/neu gene amplification. In this study, we evaluated the sensitivity and specificity of a commercially available LightCycler assay as it compares to FISH. To determine whether this assay provides an accurate alternative for the determination of HER-2/neu status, we focused primarily on tumors that were deemed indeterminate or borderline status by IHC.

Methods

Thirty-nine breast tumors receiving an IHC score of 2+ were evaluated by both FISH and LightCycler® technologies in order to determine whether quantitative real-time PCR provides an accurate alternative for the determination of HER-2/neu status.

Results

We found a high concordance (92%) between FISH and real-time PCR results. We also observed that 10% of these tumors were positive for gene amplification by both FISH and real-time PCR.

Conclusion

The data show that the results obtained for the gene amplification of HER-2/neu by real-time PCR on the LightCycler® instrument is comparable to results obtained by FISH. These results therefore suggest that real-time PCR analysis, using the LightCycler®, is a viable alternative to FISH for reassessing breast tumors which receive an IHC score of 2+, and that a combined IHC and real-time PCR approach for the determination of HER-2 status in breast cancer patients may be an effective and efficient strategy.
High sensitivity assays for docetaxel and paclitaxel in plasma using solid-phase extraction and high-performance liquid chromatography with UV detection
Authors: Andersen Anders , Warren David J , Brunsvig Paal F , Aamdal Steinar , Kristensen Gunnar B , Olsen Harald
Published: January 2006

Background

The taxanes paclitaxel and docetaxel have traditionally been used in high doses every third week in the treatment of cancer. Lately there has been a trend towards giving weekly low doses to improve the therapeutic index. This article describes the development of high performance liquid chromatographic (HPLC) methods suitable for monitoring taxane levels in patients, focusing on patients receiving low-dose therapy.

Methods

Paclitaxel and docetaxel were extracted from human plasma by solid phase extraction, and detected by absorbance at 227 nm after separation by reversed phase high performance liquid chromatography. The methods were validated and their performance were tested using samples from patients receiving paclitaxel or docetaxel.

Results

The limits of quantitation were 1 nM for docetaxel and 1.2 nM for paclitaxel. For both compounds linearity was confirmed from the limit of quantitation up to 1000 nM in plasma. The recoveries ranged between 92% and 118% for docetaxel and between 76% and 104% for paclitaxel. Accuracy and precision were within international acceptance criteria, that is within ± 15%, except at the limit of quantitation where values within ± 20% are acceptable. Low-dose patients included in an on going clinical trial had a median docetaxel concentration of 2.8 nM at 72 hours post infusion. Patients receiving 100 mg/m2 of paclitaxel had a mean paclitaxel concentration of 21 nM 48 hours after the end of infusion.

Conclusion

We have developed an HPLC method using UV detection capable of quantifying 1 nM of docetaxel in plasma samples. The method should be useful for pharmacokinetic determinations at all relevant doses of docetaxel. Using a similar methodology paclitaxel can be quantified down to a concentration of 1.2 nM in plasma with acceptable accuracy and precision. We further demonstrate that the previously reported negative influence of Cremophor EL on assay performance may be overcome by degradation of the detergent by incubation with lipase.
Effect of a brief intervention on evidence-based medicine skills of pediatric residents
Authors: Dinkevich Eugene , Markinson Andrea , Ahsan Sama , Lawrence Barbara
Published: January 2006

Background

While Evidence-Based Medicine (EBM) skills are increasingly being taught in medical schools, teaching quality has been insufficient, so that incoming pediatric residents lack adequate EBM skills required for patient care. The objective of this study was to evaluate the effectiveness of a brief teaching module developed to improve EBM skills of pediatric residents.

Methods

With-in subjects study design with pre- and post-test evaluation was performed in a large urban pediatric residency training program in Brooklyn, New York. We included PGY-1s during intern orientation, while second and third year pediatric residents were selected based on schedule availability. Sixty-nine residents were enrolled into the study, 60 (87%) completed the training. An EBM training module consisting of three or four weekly two-hour seminars was conducted. The module was designed to teach core EBM skills including (1) formulating answerable clinical questions, (2) searching the evidence, (3) critical appraisal skills including validity and applicability, and (4) understanding levels of evidence and quantitative results for therapy articles. A portion of the Fresno test of competence in EBM was used to assess EBM skills. The test presented a clinical scenario that was followed by nine short answer questions. One to three questions were used to assess EBM skills for each of the four core skills. The κ co-efficient for inter-rater reliability was 0.74 (95% CI: 0.56–0.92).

Results

Prior to the training module, the residents achieved a mean score of 17% correct overall. Post intervention, the mean score increased to 63% with improvement in each EBM category. A mean of 4.08 more questions (out of 9) were answered correctly after the training (95% CI of 3.44–4.72).

Conclusion

A brief training module was effective in improving EBM skills of pediatric residents.
Randomized controlled trial comparing four strategies for delivering e-curriculum to health care professionals [ISRCTN88148532]
Authors: Kemper Kathi J , Gardiner Paula , Gobble Jessica , Mitra Ananda , Woods Charles
Published: January 2006

Background

Internet education is increasingly provided to health professionals, but little is known about the most effective strategies for delivering the content. The purpose of this study is to compare four strategies for delivering an Internet-based (e-) curriculum on clinicians' knowledge (K), confidence (CONF), and communication (COMM) about herbs and other dietary supplements (HDS).

Methods

This national randomized 2 × 2 factorial trial included physicians, pharmacists, nurses, nutritionists and trainees in these fields. Participants were randomly assigned to one of four curriculum delivery strategies for 40 brief modules about HDS: a) delivering four (4) modules weekly over ten (10) weeks by email (drip-push); b) modules accessible on web site with 4 reminders weekly for 10 weeks (drip-pull); c) 40 modules delivered within 4 days by email (bolus-push); and d) 40 modules available on the Internet with one email informing participants of availability (bolus-pull).

Results

Of the 1,267 enrollees, 25% were male; the average age was 40 years. The completion rate was 62%, without significant differences between delivery groups. There were statistically significant improvements in K, CONF and COMM scores after the course (P<0.001 for all), although the difference in COMM was small. There were no significant differences in any of the three outcomes by delivery strategy, but outcomes were better for those who paid for continuing education credit.

Conclusion

All delivery strategies tested similarly improved K, CONF, COMM scores about HDS. Educators can use the strategy that is most convenient without diminishing effectiveness. Additional curricula may be necessary to make substantial changes in clinicians' communication practices.
Knowledge loss of medical students on first year basic science courses at the university of Saskatchewan
Authors: D'Eon Marcel F
Published: January 2006

Background

Many senior undergraduate students from the University of Saskatchewan indicated informally that they did not remember much from their first year courses and wondered why we were teaching content that did not seem relevant to later clinical work or studies. To determine the extent of the problem a course evaluation study that measured the knowledge loss of medical students on selected first year courses was conducted.

This study replicates previous memory decrement studies with three first year medicine basic science courses, something that was not found in the literature. It was expected that some courses would show more and some courses would show less knowledge loss.

Methods

In the spring of 2004 over 20 students were recruited to retake questions from three first year courses: Immunology, physiology, and neuroanatomy. Student scores on the selected questions at the time of the final examination in May 2003 (the 'test') were compared with their scores on the questions 10 or 11 months later (the 're-test') using paired samples t -tests. A repeated-measures MANOVA was used to compare the test and re-test scores among the three courses. The re-test scores were matched with the overall student ratings of the courses and the student scores on the May 2003 examinations.

Results

A statistically significant main effect of knowledge loss (F = 297.385; p < .001) and an interaction effect by course (F = 46.081; p < .001) were found. The students' scores in the Immunology course dropped 13.1%, 46.5% in Neuroanatomy, and 16.1% in physiology. Bonferroni post hoc comparisons showed a significant difference between Neuroanatomy and Physiology (mean difference of 10.7, p = .004).

Conclusion

There was considerable knowledge loss among medical students in the three basic science courses tested and this loss was not uniform across courses. Knowledge loss does not seem to be related to the marks on the final examination or the assessment of course quality by the students.
Quality research in healthcare: are researchers getting enough statistical support?
Authors: Omar Rumana Z , McNally Nick , Ambler Gareth , Pollock Allyson M
Published: January 2006

Background

Reviews of peer-reviewed health studies have highlighted problems with their methodological quality. As published health studies form the basis of many clinical decisions including evaluation and provisions of health services, this has scientific and ethical implications. The lack of involvement of methodologists (defined as statisticians or quantitative epidemiologists) has been suggested as one key reason for this problem and this has been linked to the lack of access to methodologists. This issue was highlighted several years ago and it was suggested that more investments were needed from health care organisations and Universities to alleviate this problem.

Methods

To assess the current level of methodological support available for health researchers in England, we surveyed the 25 National Health Services Trusts in England, that are the major recipients of the Department of Health's research and development (R&D) support funding.

Results and discussion

The survey shows that the earmarking of resources to provide appropriate methodological support to health researchers in these organisations is not widespread. Neither the level of R&D support funding received nor the volume of research undertaken by these organisations showed any association with the amount they spent in providing a central resource for methodological support for their researchers.

Conclusion

The promotion and delivery of high quality health research requires that organisations hosting health research and their academic partners put in place funding and systems to provide appropriate methodological support to ensure valid research findings. If resources are limited, health researchers may have to rely on short courses and/or a limited number of advisory sessions which may not always produce satisfactory results.
A blended knowledge translation initiative to improve colorectal cancer staging [ISRCTN56824239]
Authors: Wright Frances C , Law Calvin HL , Last Linda D , Klar Neil , Ryan David P , Smith Andrew J
Published: January 2006

Background

A significant gap has been documented between best practice and the actual practice of surgery. Our group identified that colorectal cancer staging in Ontario was suboptimal and subsequently developed a knowledge translation strategy using the principles of social marketing and the influence of expert and local opinion leaders for colorectal cancer.

Methods/Design

Opinion leaders were identified using the Hiss methodology. Hospitals in Ontario were cluster-randomized to one of two intervention arms. Both groups were exposed to a formal continuing medical education session given by theexpert opinion leader for colorectal cancer. In the treatment group the local Opinion Leader for colorectal cancer was detailed by theexpert opinion leader for colorectal cancerand received a toolkit. Forty-two centres agreed to have theexpert opinion leader for colorectal cancercome and give a formal continuing medical education session that lasted between 50 minutes and 4 hours. No centres refused the intervention. These sessions were generally well attended by most surgeons, pathologists and other health care professionals at each centre. In addition all but one of the localopinion leaders for colorectal cancermet with theexpert opinion leader for colorectal cancerfor the academic detailing session that lasted between 15 and 30 minutes.

Discussion

We have enacted a unique study that has attempted to induce practice change among surgeons and pathologists using an adapted social marketing model that utilized the influence of both expert and local opinion leaders for colorectal cancer in a large geographic area with diverse practice settings.