bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR
Authors: Del Aguila Eduardo M , Dutra Marcio B , Silva Joab T , Paschoalin Vânia MF 
Published: April 2005

Abstract
Background
Preparation of RNA free from DNA is a critical step before performing RT-PCR assay. Total RNA isolated from several sources, including those obtained from Saccharomyces cerevisiae, using routine methodologies are frequently contaminated with DNA, which can give rise to amplification products that mimic the amplicons expected from the RNA target.

Results
We investigated the efficiency of two DNase I based protocols for eliminating DNA contaminations from RNA samples obtained from yeast cells. Both procedures are very efficient in eliminating DNA contamination from RNA samples and entail three main steps, which involve treating of RNA samples with DNase I, inhibition of the enzyme by EDTA and its subsequent inactivation at 65°C. The DNase I treated samples were further purified with phenol: chloroform followed by precipitation with ice-cold ethanol (protocol I) or, alternatively, they were directly used in RT-PCR reactions (protocol II). Transcripts from ACT1, PDA1, CNA1, CNA2, TPS1 and TPS2 analyzed after each treatment showed that all mRNAs tested can be amplified if total RNA was extracted and purified after DNase I treatment, however, only TPS1, TPS2 and ACT1 mRNAs were amplified without extraction/purification step.

Conclusion
Although more laborious and requiring a higher initial amount of material, the inclusion of an extraction and purification step allows to prepare RNA samples that are free from DNA and from low molecular contaminants and can be applied to amplify any Saccharomyces cerevisiae mRNA by RT-PCR.
Identifying future models for delivering genetic services: a nominal group study in primary care
Authors: Elwyn Glyn , Edwards Adrian , Iredale Rachel , Davies Peter , Gray Jonathon 
Published: April 2005

BackgroundTo enable primary care medical practitioners to generate a range of possible service delivery models for genetic counselling services and critically assess their suitability.
Methods
Modified nominal group technique using in primary care professional development workshops.

Results
37 general practitioners in Wales, United Kingdom too part in the nominal group process. The practitioners who attended did not believe current systems were sufficient to meet anticipated demand for genetic services. A wide range of different service models was proposed, although no single option emerged as a clear preference. No argument was put forward for genetic assessment and counselling being central to family practice, neither was there a voice for the view that the family doctor should become skilled at advising patients about predictive genetic testing and be able to counsel patients about the wider implications of genetic testing for patients and their family members, even for areas such as common cancers. Nevertheless, all the preferred models put a high priority on providing the service in the community, and often co-located in primary care, by clinicians who had developed expertise.

Conclusion
There is a need for a wider debate about how healthcare systems address individual concerns about genetic concerns and risk, especially given the increasing commercial marketing of genetic tests.
Cerebral infarction in diabetes: Clinical pattern, stroke subtypes, and predictors of in-hospital mortality
Authors: Arboix Adrià , Rivas Antoni , García-Eroles Luis , de Marcos Lourdes , Massons Joan , Oliveres Montserrat 
Published: April 2005

Abstract
Background
To compare the characteristics and prognostic features of ischemic stroke in patients with diabetes and without diabetes, and to determine the independent predictors of in-hospital mortality in people with diabetes and ischemic stroke.

Methods
Diabetes was diagnosed in 393 (21.3%) of 1,840 consecutive patients with cerebral infarction included in a prospective stroke registry over a 12-year period. Demographic characteristics, cardiovascular risk factors, clinical events, stroke subtypes, neuroimaging data, and outcome in ischemic stroke patients with and without diabetes were compared. Predictors of in-hospital mortality in diabetic patients with ischemic stroke were assessed by multivariate analysis.

Results
People with diabetes compared to people without diabetes presented more frequently atherothrombotic stroke (41.2% vs 27%) and lacunar infarction (35.1% vs 23.9%) (P < 0.01). The in-hospital mortality in ischemic stroke patients with diabetes was 12.5% and 14.6% in those without (P = NS). Ischemic heart disease, hyperlipidemia, subacute onset, 85 years old or more, atherothrombotic and lacunar infarcts, and thalamic topography were independently associated with ischemic stroke in patients with diabetes, whereas predictors of in-hospital mortality included the patient's age, decreased consciousness, chronic nephropathy, congestive heart failure and atrial fibrillation

Conclusion
Ischemic stroke in people with diabetes showed a different clinical pattern from those without diabetes, with atherothrombotic stroke and lacunar infarcts being more frequent. Clinical factors indicative of the severity of ischemic stroke available at onset have a predominant influence upon in-hospital mortality and may help clinicians to assess prognosis more accurately.
Comparison of SPECT bone scintigraphy with MRI for diagnosis of meniscal tears
Authors: Tahmasebi Mohammad-naghi , Saghari Mohsen , Moslehi Masoud , Gholamrezanezhad Ali 
Published: April 2005

AbstractBackgroundScintigraphy has been considered as competitive to MRI, but limited data are available on the accuracy of single photon emission tomography (SPECT) compared with MRI for the assessment of meniscal tears. Our objective was to assess the value of SPECT in comparison to MRI.MethodsBetween January 2003 and March 2004, sixteen patients were studied with both modalities and the accuracy rates of SPECT scan results, and MRI findings in the diagnosis of meniscal tears were compared. Arthroscopy was the gold standard.ResultsThe respective sensitivity rate, specificity rate, and positive and negative predictive accuracies of MRI were 89%, 94%, 93%, and 79% and for SPECT those were 78%, 94%, 94%, and 88%. There was good agreement on the presence or absence of tears between two modalities (κ statistic = 0.699).ConclusionSPECT and MRI are both valuable imaging techniques. SPECT is a useful alternative when MRI is unavailable or unsuitable and it is beneficial when more possible accuracy is desired (such as when MRI results are either inconclusive or conflict with other clinical data).
Rare mutations predisposing to familial adenomatous polyposis in Greek FAP patients
Authors: Mihalatos Markos , Apessos Angela , Dauwerse Hans , Velissariou Voula , Psychias Aristidis , Koliopanos Alexander , Petropoulos Konstantinos , Triantafillidis John K , Danielidis Ioannis , Fountzilas George , Agnantis Niki J , Nasioulas Georgios 
Published: April 2005

Abstract

Background
Familial Adenomatous Polyposis (FAP) is caused by germline mutations in the APC (Adenomatous Polyposis Coli) gene. The vast majority of APC mutations are point mutations or small insertions / deletions which lead to truncated protein products. Splicing mutations or gross genomic rearrangements are less common inactivating events of the APC gene.

Methods
In the current study genomic DNA or RNA from ten unrelated FAP suspected patients was examined for germline mutations in the APC gene. Family history and phenotype were used in order to select the patients. Methods used for testing were dHPLC (denaturing High Performance Liquid Chromatography), sequencing, MLPA (Multiplex Ligation – dependent Probe Amplification), Karyotyping, FISH (Fluorescence In Situ Hybridization) and RT-PCR (Reverse Transcription – Polymerase Chain Reaction).

Results
A 250 Kbp deletion in the APC gene starting from intron 5 and extending beyond exon 15 was identified in one patient. A substitution of the +5 conserved nucleotide at the splice donor site of intron 9 in the APC gene was shown to produce frameshift and inefficient exon skipping in a second patient. Four frameshift mutations (1577insT, 1973delAG, 3180delAAAA, 3212delA) and a nonsense mutation (C1690T) were identified in the rest of the patients.

Conclusion
Screening for APC mutations in FAP patients should include testing for splicing defects and gross genomic alterations.
Immune function biomarkers in children exposed to lead and organochlorine compounds: a cross-sectional study
Authors: Karmaus Wilfried , Brooks Kevin R , Nebe Thomas , Witten Jutta , Obi-Osius Nadia , Kruse Hermann 
Published: April 2005

Abstract
Background
Different organochlorines and lead (Pb) have been shown to have immunomodulating properties. Children are at greater risk for exposure to these environmental toxicants, but very little data exist on simultaneous exposures to these substances.

Methods
We investigated whether the organochlorine compounds (OC) dichlorodiphenylethylene (DDE), hexachlorobenzene (HCB), hexachlorocyclohexane (γ-HCH), the sum of polychlorinated biphenyls (ΣPCBs) and Pb were associated with immune markers such as immunoglobulin (Ig) levels, white blood cell (WBC), counts of lymphocytes; eosinophils and their eosinophilic granula as well as IgE count on basophils. The investigation was part of a cross-sectional environmental study in Hesse, Germany. In 1995, exposure to OC and Pb were determined, questionnaire data collected and immune markers quantified in 331 children. For the analyses, exposure (OC and Pb) concentrations were grouped in quartiles (γ-HCH into tertiles). Using linear regression, controlling for age, gender, passive smoking, serum lipids, and infections in the previous 12 months, we assessed the association between exposures and immune markers. Adjusted geometric means are provided for the different exposure levels.

Results
Geometric means were: DDE 0.32 μg/L, ΣPCBs 0.50 μg/L, HCB 0.22 μg/L, γ-HCH 0.02 μg/L and Pb 26.8 μg/L. The ΣPCBs was significantly associated with increased IgM levels, whereas HCB was inversely related to IgM. There was a higher number of NK cells (CD56+) with increased γ-HCH concentrations. At higher lead concentrations we saw increased IgE levels. DDE showed the most associations with significant increases in WBC count, in IgE count on basophils, IgE, IgG, and IgA levels. DDE was also found to significantly decrease eosinophilic granula content.

Conclusion
Low-level exposures to OC and lead (Pb) in children may have immunomodulating effects. The increased IgE levels, IgE count on basophils, and the reduction of eosinophilic granula at higher DDE concentrations showed a most consistent pattern, which could be of clinical importance in the etiology of allergic diseases.
Skin testing versus radioallergosorbent testing for indoor allergens
Authors: Chinoy Birjis , Yee Edgar , Bahna Sami L 
Published: April 2005

AbstractBackgroundSkin testing (ST) is the most common screening method for allergy evaluation. Measurement of serum specific IgE is also commonly used, but less so by allergists than by other practitioners. The sensitivity and specificity of these testing methods may vary by type of causative allergen and type of allergic manifestation. We compared ST reactivity with serum specific IgE antibodies to common indoor allergens in patients with respiratory allergies.Methods118 patients (3 mo-58 yr, mean 12 yr) with allergic rhinitis and/or bronchial asthma had percutaneous skin testing (PST) supplemented by intradermal testing (ID) with those allergens suspected by history but showed negative PST. The sera were tested blindly for specific IgE antibodies by the radioallergosorbent test (Phadebas RAST). The allergens wereD. farinae(118), cockroach (60), cat epithelium (90), and dog epidermal (90). Test results were scored 0–4; ST ≥ 2 + and RAST ≥ 1 + were considered positive.ResultsThe two tests were in agreement (i.e., either both positive or both negative) in 52.2% (dog epidermal) to 62.2% (cat epithelium). When RAST was positive, ST was positive in 80% (dog epidermal) to 100% (cockroach mix). When ST was positive, RAST was positive in 16.3% (dog epidermal) to 50.0%(D. farinae). When RAST was negative, ST was positive in 48.5% (cat epithelium) to 69.6% (D. farinae). When ST was negative, RAST was positive in 0% (cockroach) to 5.6% (cat epithelium). The scores of ST and RAST showed weak to moderate correlation (r = 0.24 to 0.54). Regardless of history of symptoms on exposure, ST was superior to RAST in detecting sensitization to cat epithelium and dog epidermal.ConclusionFor all four indoor allergens tested, ST was more sensitive than RAST. When both tests were positive, their scores showed poor correlation. Sensitizations to cat epithelium and dog epidermal are common, even in subjects who claimed no direct exposure.
Educational and labor wastage of doctors in Mexico: towards the construction of a common methodology
Authors: Nigenda Gustavo , Ruiz José Arturo , Bejarano Rosa 
Published: April 2005

Abstract
Background
This paper addresses the problem of wastage of the qualified labor force, which takes place both during the education process and when trained personnel try to find jobs in the local market.

Methods
Secondary sources were used, mainly the Statistical yearbooks of the National Association of Universities and Higher Education Institutions (ANUIES in Spanish). Also, the 2000 Population Census was used to estimate the different sources of labor market wastage. The formulas were modified to estimate educational and labor wastage rates.

Results
Out of every 1000 students who started a medical training in 1996, over 20% were not able to finish the training by 2000. Furthermore, out of every 1000 graduates, 31% were not able to find a remunerated position in the labor market that would enable them to put into practice the abilities and capacities obtained at school. Important differences can be observed between generalists and specialists, as well as between men and women. In the case of specialists and men, lower wastage rates can be observed as compared to the wastage rates of generalists and women. A large percentage of women dedicate themselves exclusively to household duties, which in labor terms represents a wastage of their capacity to participate in the production of formal health services.

Conclusion
Women are becoming a majority in most medical schools, yet their participation in the labor market does not reflect the same trend. Among men, policies should be formulated to incorporate doctors in the specific health field for which they were trained. Regarding women, specific policies should target those who are dedicated full-time to household activities in order to create the possibility of having them occupy a remunerated job if they are willing to do so. Reducing wastage at both the educational and labor levels should improve the capacity of social investment, thereby increasing the capacity of the health system as a whole to provide services, particularly to those populations who are most in need.
Comparable response of ccn1 with ccn2 genes upon arthritis: An in vitro evaluation with a human chondrocytic cell line stimulated by a set of cytokines
Authors: Moritani Norifumi H , Kubota Satoshi , Sugahara Toshio , Takigawa Masaharu 
Published: April 2005

Abstract

Background
The chondrosarcoma-derived HCS-2/8 has been known to be an excellent model of human articular chondrocytes. By mimicking the arthritic conditions through the treatment of HCS-2/8 cells with cytokines, we estimated the gene expression response of ccn1 and ccn2 during the course of joint inflammation in vitro.

Results
In order to mimic the initiation of inflammation, HCS-2/8 cells were treated with tumor necrosis factor (TNF)-α. To induce pro-inflammatory or reparative responses, TGF-β was employed. Effects of an anti-inflammatory glucocorticoid were also evaluated. After stimulation, expression levels of ccn1 and ccn2 were quantitatively analyzed. Surprisingly, not only ccn2, but also ccn1 expression was repressed upon TNF-α stimulation, whereas both mRNAs were uniformly induced by transforming growth factor (TGF)-β and a glucocorticoid.

Conclusion
These results describing the same response during the course of inflammation suggest similar and co-operative roles of these 2 ccn family members in the course of arthritis.
Eco-epidemiological analysis of dengue infection during an outbreak of dengue fever, India
Authors: Chakravarti Anita , Kumaria Rajni 
Published: April 2005

Abstract

Background
This study was designed to find out a relationship of dengue infection with climatic factors such as rainfall, temperature and relative humidity during the dengue fever epidemic in the year 2003. Blood samples were collected from 1550 patients experiencing a febrile illness clinically consistent with dengue infection. Serological confirmation of Dengue Infection was done using Dengue Duo IgM and IgG Rapid Strip test (Pan Bio, Australia), which detected dengue-specific antibodies. Monthly data of total rainfall, temperature and relative humidity for the year 2003 was obtained from Meteorological Department of Delhi, New Delhi and retrospectively analyzed.

Results
Out of 1550 suspected cases, 893 cases (57.36%) were confirmed as serologically positive. The difference between numbers of serologically positive cases during different months was significant (p < 0.05). Larger proportions of serologically positive cases were observed among adults. Outbreak coincided mainly with the post monsoon period of subnormal rainfall. The difference between serologically positive cases as compared to serologically negative ones in post monsoon period was significantly higher (p < 0.001). The difference in the rainfall and temperature between three seasonal periods was significant (p < 0.05).

Conclusion
This prospective study highlighted rain, temperature and relative humidity as the major and important climatic factors, which could alone or collectively be responsible for an outbreak. More studies in this regard could further reveal the correlation between the climatic changes and dengue outbreaks, which would help in making the strategies and plans to forecast any outbreak in future well in advance.
Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein
Authors: Marmey Philippe , Rojas-Mendoza Ana , de Kochko Alexandre , Beachy Roger N , Fauquet Claude M 
Published: April 2005

Abstract

Background
Rice tungro bacilliform virus (RTBV) is a pararetrovirus, and a member of the family Caulimoviridae in the genus Badnavirus. RTBV has a long open reading frame that encodes a large polyprotein (P3). Pararetroviruses show similarities with retroviruses in molecular organization and replication. P3 contains a putative movement protein (MP), the capsid protein (CP), the aspartate protease (PR) and the reverse transcriptase (RT) with a ribonuclease H activity. PR is a member of the cluster of retroviral proteases and serves to proteolytically process P3. Previous work established the N- and C-terminal amino acid sequences of CP and RT, processing of RT by PR, and estimated the molecular mass of PR by western blot assays.

Results
A molecular mass of a protein that was associated with virions was determined by in-line HPLC electrospray ionization mass spectral analysis. Comparison with retroviral proteases amino acid sequences allowed the characterization of a putative protease domain in this protein. Structural modelling revealed strong resemblance with retroviral proteases, with overall folds surrounding the active site being well conserved. Expression in E. coli of putative domain was affected by the presence or absence of the active site in the construct. Analysis of processing of CP by PR, using pulse chase labelling experiments, demonstrated that the 37 kDa capsid protein was dependent on the presence of the protease in the constructs.

Conclusion
The findings suggest the characterization of the RTBV protease domain. Sequence analysis, structural modelling, in vitro expression studies are evidence to consider the putative domain as being the protease domain. Analysis of expression of different peptides corresponding to various domains of P3 suggests a processing of CP by PR. This work clarifies the organization of the RTBV polyprotein, and its processing by the RTBV protease.
Molecular advances in the cell biology of SARS-CoV and current disease prevention strategies
Authors: Stark Caren J , Atreya CD 
Published: April 2005

Abstract
In the aftermath of the SARS epidemic, there has been significant progress in understanding the molecular and cell biology of SARS-CoV. Some of the milestones are the availability of viral genome sequence, identification of the viral receptor, development of an infectious cDNA clone, and the identification of viral antigens that elicit neutralizing antibodies. However, there is still a large gap in our understanding of how SARS-CoV interacts with the host cell and the rapidly changing viral genome adds another variable to this equation. Now the SARS-CoV story has entered a new phase, a search for preventive strategies and a cure for the disease. This review highlights the progress made in identifying molecular aspects of SARS-CoV biology that is relevant in developing disease prevention strategies. Authors conclude that development of successful SARS-CoV vaccines and antivirals depends on the progress we make in these areas in the immediate future.
Characteristics of patients with organic brain syndromes : A cross-sectional 2-year follow-up study in Kuala Lumpur, Malaysia
Authors: Chandrasekaran Prem K , Jambunathan Stephen T , Zainal Nor Z 
Published: April 2005

Abstract
Background
Organic Brain Syndromes (OBS) are often missed in clinical practice. Determining their varied presentations may help in earlier detection, better management, and, assessing prognosis and outcome. We described the in-patient referrals of patients suffering from the psychiatric effects of organic states and compared the symptomatology and mortality between those with the Acute and Chronic varieties.

Methods
59 patients referred to our Consultation-Liaison (C-L) Psychiatry services and given a clinical diagnosis of OBS were selected over a 6-month period. Psychiatric and cognitive abnormalities and treatment regimes were recorded and fatality rates determined. Information regarding their condition 24 months after their index hospitalization was recorded. All data were entered into a proforma and analyzed after exclusion.

Results
The mean duration of detecting the symptoms by the physician was 3.52 days. The presence of a premorbid psychiatric illness had no influence on the clinical presentation but did on the mortality of patients with OBS (p = 0.029).
Patients with the Acute syndrome had significantly more symptom resolution as compared to those with the Chronic syndrome (p = 0.001) but mortalityrates did not differ. Elderly patients and those with symptom resolution upon discharge did not show statistically significant higher mortality rates. The most popular combination of treatment was that of a low-dose neuroleptic and a benzodiazepine (34.7%). The need for maintenance treatment was not significantly different in any group, even in those with a past history of a functional disorder.

Conclusion
Other than the Acute group having a significantly better outcome in terms of symptom resolution, our findings suggest that there was no significant difference in the clinical presentation between those with Acute or Chronic OBS. Mortality-wise, there was also no difference between the Acute and Chronic syndromes, nor was there any difference between the elderly and the younger group. There was also no significant difference in the need for continued treatment in both groups.