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1.  An unusual unifocal presentation of Castleman’s disease in a young woman with a detailed description of sonographic findings to reduce diagnostic uncertainty: a case report 
BMC Research Notes  2013;6:97.
Background
Castleman’s disease is a rare lymphoproliferative disorder. It typically presents as mediastinal masses and causes a wide range of clinical symptoms. Histologically, Castleman’s disease is classified as either a hyalinic vascular or plasma cell variant. The prognosis mainly depends on the histological type and broadly varies. We herein report our sonographic findings in a patient with Castleman’s disease, including gray-scale ultrasonography, color Doppler ultrasonography, and sonoelastography ultrasonography, which have not been previously reported in the literature. These findings allowed for a preoperative diagnosis and avoidance of overly aggressive therapy.
Case presentation
A 28-year-old European female patient with unicentric Castleman’s disease of hyalinic vascular type (HV) restricted to the axilla was referred to us because of a 4-month history of a painless, solitary mass located in the left axilla. The patient’s medical history was unremarkable.
Conclusion
Castleman’s disease is a pathologic entity of unknown etiology and pathogenesis. In this case report of unicentric HV-type CD, we demonstrate that typical sonographic findings can lead to a preoperative diagnosis of Castleman’s disease. Core needle biopsy usually allows for a final diagnosis and helps to avoid unnecessary operations and overtreatment.
doi:10.1186/1756-0500-6-97
PMCID: PMC3601982  PMID: 23497665
Castleman’s disease; Giant lymph node hyperplasia; Ultrasonography; Core needle biopsy
2.  Women’s expectation of partner’s violence on HIV disclosure for prevention of mother to child transmission of HIV in North West Ethiopia 
BMC Research Notes  2013;6:96.
Background
All violence against women has serious consequences for their mental, physical wellbeing, reproductive and sexual health including HIV infection and no study was conducted in this regard in Ethiopia and particularly in the present study area.
Findings
A cross-sectional study was conducted in Gondar town from 22 July–18 August 2011. Of the 400 pregnant women who actively participated in this study, 314 (78.50%) expected a negative reaction for HIV positive test result from their partners. A positive reaction from the partner was associated with women having their own income (Adjusted odds ratio (AOR) (95% CI) =2.18 (1.21, 3.92)), residing in the urban areas (AOR (95% CI) =2.26 (1.21, 4.22)), having education level of secondary level and above (AOR (95% CI) = 6.05 (3.12, 11.72)), not having a stigmatizing attitude towards people living with HIV (AOR (95% CI) = 2.15 (1.24, 3.73)), having a positive attitude towards counselors (AOR (95% CI) = 2.46 (1.42, 4.25)) and being able to access health facilities (AOR (95% CI) = 2.35(1.22, 4.50)).
Conclusion
Most of the participants in this study expected their partner to react negatively towards a positive HIV test result. Since women’s having their own income is strongly associated with a positive partner’s reaction on HIV test disclosure for prevention of mother to child transmission of HIV services, emphasis should be given for education and economic empowerment of women. A well functioning and accessible health facility with prevention of mother to child transmission of HIV service is important, especially in rural areas.
doi:10.1186/1756-0500-6-96
PMCID: PMC3601008  PMID: 23497642
Male partner violence; PMTCT; Pregnant women
3.  Validation of reference genes for expression analysis by quantitative real-time PCR in Leptinotarsa decemlineata (Say) 
BMC Research Notes  2013;6:93.
Background
L. decemlineata is an exotic invasive insect pest, and invaded in Xinjiang Uygur autonomous region in China in the 1990s from Kazakhstan. It is a notorious defoliator of potato throughout most of the northern Xinjiang in current, and often causes extremely large yield losses of potato.
Results
The expression stability of nine L. decemlineata house-keeping genes (Actin, ACT1 and ACT2; ADP-ribosylation factor, ARF1 and ARF4; TATA box binding protein, TBP1 and TBP2; ribosomal protein RP4 and RP18; translation elongation factor 1α EF1α) was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) in seven developmental stages, three larval tissues and two insecticide treatments. The results were analyzed using three software programs: geNorm, NormFinder and BestKeeper. Although there was no consistent ranking observed among the house-keeping genes across the samples, the overall analysis revealed that RP18, RP4, ARF1, and ARF4 were the four most stable house-keeping genes. In contrast, ACT1 and ACT2, two of the most widely used reference genes, had the least stability. Our results suggest that the combined use of the four most stably expressed genes may produce optimal normalization for qRT-PCR.
Conclusions
The expression stability of the house-keeping genes varies among different developing stages, in different tissues and under different experimental conditions. Our results will enable a more accurate and reliable normalization of qRT-PCR data in L. decemlineata.
doi:10.1186/1756-0500-6-93
PMCID: PMC3600673  PMID: 23497596
L. decemlineata; Quantitative real-time PCR; Reference gene; Normalization
4.  Peritoneal tuberculosis mimicking advanced ovarian carcinoma: an important differential diagnosis to consider 
BMC Research Notes  2013;6:88.
Background
Female patients who present with ascites, adnexal masses and elevated CA125 levels are typically presumed to have advanced ovarian carcinoma. This can lead to radical surgery with its associated morbidity. An important differential diagnosis to consider is tuberculous peritonitis which can present in a similar manner and responds well to medical treatment.
Case presentation
A 44 year old female presented with abdominal distension, weight loss and low grade fever. Her CA125 level was 909 U/ml. Imaging studies revealed an adnexal lesion and ascites. The lungs appeared normal and a Mantoux test was negative. Ovarian malignancy was highly suspected. Cytology of ascites was negative for malignant cells. The patient subsequently developed a large pleural effusion which was drained and negative for malignant cells and acid fast bacilli. Repeat imaging revealed a ‘tree in bud’ appearance of the lung parenchyma and dense ascites. Histology from diagnostic laparotomy revealed caseating granulomas with epithelioid cells and Langhan’s type giant cells. The patient responded well to antituberculosis therapy with normalization of CA125 levels, confirming the diagnosis of peritoneal tuberculosis.
Conclusion
CA125 levels lack specificity, with elevated levels encountered in many benign and malignant conditions, including tuberculosis. There are a few discriminating features that suggest a diagnosis of tuberculous peritonitis rather than ovarian carcinoma. Apart from chest findings which may not always be present, smooth peritoneal thickening and a dirty omentum on CT favours a diagnosis of peritoneal tuberculosis compared with nodular thickening of the peritoneum and omentum in peritoneal carcinomatosis. PCR and ADA testing of ascitic fluid can also be helpful. When these tests are negative or unavailable then diagnostic laparoscopy or laparotomy should be performed with the aid of frozen section to avoid unnecessary radical surgery in cases of peritoneal tuberculosis.
doi:10.1186/1756-0500-6-88
PMCID: PMC3599504
Abdominal disseminated tuberculosis; Mimic ovarian carcinoma; CA125; Ascites; Adnexal mass
5.  Morphological and metabolic determinants of nonalcoholic fatty liver disease in obese youth: a pilot study 
BMC Research Notes  2013;6:89.
Background
Nonalcoholic fatty liver disease (NAFLD) related to obesity has been rising in the last decades, though the morphological and metabolic determinants are remain unclear in children. The aim of this study was to analyze the morphological determinants and metabolic abnormalities in obese children and adolescents, classified either as with (P-NAFLD) or without (N-NAFLD). The sample comprised 190 individuals, aged 6 to 16 years-old, assigned into one of 4 groups according to sex and presence or absence of NAFLD. Obesity was obtained according to body mass index (BMI) cut-points. Body composition variables was estimated by Dual-Energy X-ray Absorptiometry (DEXA). Total cholesterol (TC), triglycerides (TG), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), insulin, fasting glucose (FG) and blood pressure were also analyzed. The diagnosis of NAFLD, as well as the measurement of intra-abdominal fat tissue thickness (IAF) and subcutaneous abdominal fat tissue thickness (SCF), was carried-out by ultrasound.
Results
Males and females belonging to P-NAFLD group showed, respectively, higher TFM and IAF. When data were adjusted for sex, age and total fat mass, those in P-NAFLD showed statistically higher IAF, TFM and TG.
Conclusion
Our study showed that obese youngsters who were assigned to P-NAFLD group were twice as likely to present higher concentration of triglycerides, higher levels of trunk fat, as well as intra-abdominal fat compared to their N-NAFLD counterparts even after adjustments for sex, age, pubertal stage and total body fat mass.
doi:10.1186/1756-0500-6-89
PMCID: PMC3599575  PMID: 23497552
Adolescents; Children; Nonalcoholic fatty liver disease; Obesity
6.  Stair descending exercise increases muscle strength in elderly males with chronic heart failure 
BMC Research Notes  2013;6:87.
Background
Previous studies from our group have shown that "pure" eccentric exercise performed on an isokinetic dynamometer can induce health-promoting effects that may improve quality of life. In order to investigate whether the benefits of "pure" eccentric exercise can be transferred to daily activities, a new and friendlier way to perform eccentric exercise had to be invented. To this end, we have proceeded to the design and construction of an automatic escalator, offering both stair descending (eccentric-biased) and stair ascending (concentric-biased) exercise.
Findings
Twelve elderly males (60-70 yr) with chronic heart failure participated in the present study. Participants carried out six weeks of stair descending or ascending training on the novel SmartEscalator device. Muscle damage and performance indices were evaluated before and at day 2 post exercise at the first and sixth week of training. Both training regimes increased, albeit not significantly in some cases, eccentric, concentric and isometric torque. After six weeks of stair descending exercise, eccentric, concentric and isometric peak torque increased 12.3%, 7.7% and 8.8%, respectively, whereas after stair ascending exercise eccentric, concentric and isometric peak torque increased 7.1%, 9.6% and 5.9%, respectively.
Conclusions
Stair descending exercise appears to be a pleasant and mild activity that can be easily followed by the elderly. Compared to the more demanding stair ascending exercise, changes in muscle strength are similar or even greater. Elderly or people with impaired endurance wishing to increase their muscle strength may be benefited by participating in activities with strong eccentric component, such as stair descending.
doi:10.1186/1756-0500-6-87
PMCID: PMC3601017
Aging; Chronic heart failure; Eccentric exercise; Muscle strength; Sarcopenia
7.  School-based injury outcomes in children from a low-income setting: results from the pilot injury surveillance in Rawalpindi city, Pakistan 
BMC Research Notes  2013;6:86.
Background
School-based injuries account for one in five unintentional childhood injuries. Little is known about the epidemiology of school-based injuries in low-income settings. The objective of our study was to compare emergency department (ED) outcomes of the school-based injuries with respect to age, sex, and injury mechanisms in a Pakistani urban setting.
Findings
A pilot injury surveillance study was conducted at the EDs of three major tertiary-care hospitals of Rawalpindi city from July 2007 to June 2008 and included children of less than 15 years injured at school. The World Health Organization’s questionnaire for injury surveillance was used. There were 923 school injury cases. Mean age of children involved was 8.3 years (SD ± 3.3) with male female ratio 2.9:1. Most injuries occurred while playing 85.6% (n = 789); of which the most common mechanism was falls (n = 797, 86.4%). Nineteen of twenty cases were directly discharged home from the ED (N = 861). Compared to ED discharged cases, injury characteristics overrepresented in hospital admitted cases (n = 46) were age 10–14 years (65.2% vs. 40.9%, p = 0.005), male (88.6% vs. 25.9%), involved in educational activities (39.1% vs. 5.3%), injured from fire/heat (37.8% vs. 0.6%), had burns (39.5% vs. 0.9%) and head injuries (27.9% vs. 6.4%).
Conclusion
Falls while playing are the commonest injury mechanism in school-based injuries reported in our ED sample. School officials need to prevent these injuries. Studying injury hazards present in school environment in Pakistan might facilitate developing specific prevention strategies.
doi:10.1186/1756-0500-6-86
PMCID: PMC3599831
School injuries; Children; Pakistan
8.  Lower ratio of high-molecular-weight adiponectin level to total may be associated with coronary high-risk plaque 
BMC Research Notes  2013;6:83.
Background
Although high-molecular-weight (HMW) adiponectin is believed to protect against atherosclerosis, the association between HMW adiponectin and the composition of coronary plaques is unknown. We evaluated whether the HMW to total adiponectin ratio was associated with the presence of coronary plaque and its composition using multi-slice computed tomography coronary angiography (MSCTCA).
Methods
Serum total and HMW adiponectin levels were measured in 53 consecutive patients (age, 71) with >50% coronary artery stenosis detected by MSCTCA. A low-attenuation coronary plaque was defined as a plaque with a mean CT density <50 Hounsfield units. Multivariate logistic regression analyses were performed to evaluate the predictors of the presence of low-attenuation coronary plaques, which is thought to be high risk, on CT.
Results
Decreased serum levels of total as well as HMW adiponectin were significantly associated with the presence of at least one calcified or non-calcified coronary artery plaque (total adiponectin level: odds ratio 0.76, 95% CI 0.58–0.99, P = 0.048; HMW adiponectin level: odds ratio 0.65, 95% CI 0.42–0.99, P = 0.047). A low ratio of HMW to total adiponectin was significantly associated with the presence of low-attenuation coronary plaques (4.55, 1.94–21.90, P = 0.049). However, neither the total adiponectin nor the HMW adiponectin level was associated with the presence of low-attenuation coronary plaques.
Conclusion
Lower total or HMW adiponectin levels are associated with the presence of calcified and non-calcified coronary plaques, whereas a lower ratio of HMW to total adiponectin associated with the presence of low-attenuation coronary plaques (thought to be high risk). Measurement of total and HMW adiponectin levels and the HMW to total adiponectin ratio may be useful for risk stratification of coronary artery plaques.
doi:10.1186/1756-0500-6-83
PMCID: PMC3599310  PMID: 23497474
Adiponectin; High-molecular-weight adiponectin; Coronary artery plaque; Coronary low-attenuation plaque
9.  Correlation between the operation time using two different power settings of a Ho: YAG laser: laser power doesn’t influence lithotripsy time 
BMC Research Notes  2013;6:80.
Background
This study investigated the correlation between the operation time using two different power settings of a Ho: YAG laser.
Findings
A total of 68 patients underwent cystolithotripsy from April 2010 to October 2011 In Fifty-six of these patients underwent cystolithotripsy by one surgeon using a Ho: YAG laser for bladder calculi. This study assessed these patients in two groups; the 30 W laser generator group with the settings of 2.5 J x 5 Hz (30 W group) and the 100 W laser generator group as the settings of 3.5 J x 5 Hz (100 W group). The operation time in these two groups were assessed.
A total of 56 patients including 45 male and 11 female patients that underwent cystolithotripsy using a Ho: YAG laser for bladder calculi by one surgeon were enrolled in this study. The patients’ characteristics including age (mean; 68.8 vs 68.4 yr), gender (male; 74.2 vs 88.0%), stone burden (mean; 34.9 vs 41.3 mm), number of stones (mean; 3.2 vs 2.0) and stone’s CT density (mean; 981.5 vs 902.0 HU) showed no significant differences. All patients were stone free following treatment. The median total length of the operation was 19 minutes (mean: 34.6 ± 36.1) in the 30 W group and 29 minutes (mean: 44.4 ± 38.8) in the 100 W group, which was not significantly different.
Conclusions
The results showed that the power settings of Ho: YAG laser show no differences in the operation time for bladder calculi lithotripsy.
doi:10.1186/1756-0500-6-80
PMCID: PMC3599216
Cystolithotripsy; Ho; YAG laser; Bladder stone; Bladder calculi
10.  Pichia fabianii blood infection in a premature infant in China: case report 
BMC Research Notes  2013;6:77.
Background
Invasive fungal infections caused by uncommon fungi have increased in recent years. Hospitalized low-birth-weight infants are at high risk for neonatal fungal infections. Pichia fabianii is a rare pathogen causing blood infection, which has reportedly caused only 4 cases of fungemia and 1 case of endocarditis worldwide. Here, we describe the first case of a P. fabianii blood infection in a premature infant in China.
Case presentation
On July 28th, a low-birth-weight (LBW, 1760 g) female infant born at 33+4 weeks of gestation was admitted to the pediatric intensive care unit with mild neonatal asphyxia. Until August 2nd, a mechanical respirator was used to assist respiration under the Continuous Positive Airway Pressure (CPAP) model. The baby had an increased body temperature and a fever. To prevent infection, Ceftriaxone Sodium (CS) was administered intravenously for three days, after which Cefepime was administered until August 13th. Chest X-rays showed suspected plaque-like shadows in the right lung. Blood cultures twice tested positive for fungal infection caused by Candida pelliculosa (recognized as Pichia fabianii later), which is first mis-identified by commercial kit. Hence, intravenous fluconazole was administered. However, cultures of other body fluids (e.g., urine, feces and sputum) tested negative for fungal infection. Routine tests and biochemistry of cerebrospinal fluid (CSF) were normal. Latex agglutination of Cryptococcus neoformans and fungi cultures in the CSF were also negative. After 14 days of intravenous fluconazole, blood was re-cultured, the result of which was negative. On August 30th, intravenous fluconazole was suspended. On Sep 3rd, the infant left the hospital in good health.
Conclusions
This is the first case of a blood infection caused by P. fabianii in a LBW premature female infant in China. Risk factors for fungal infection include premature birth, as well as mechanical invasive operation and antibacterial drug usage. Whether such risk factors necessitate prophylactic use of antifungal drugs is an important question that has yet to be fully addressed. Additionally, the pathogen P. fabianii collected in this study was resistant to amphotericin B (AMB) and itraconazole (ITR). With the exception of the azole-resistant endocarditis case, all other cases have not demonstrated such a resistance. Finally, commercial biochemical methods used in routine practice are limited in their ability to identify P. fabianii. Molecular genetic based methods are imperative for identification of uncommon fungal species from disseminated infections.
doi:10.1186/1756-0500-6-77
PMCID: PMC3599298
Pichia fabianii; Fungemia; Infant infection
11.  Aerobic exercise training and burnout: a pilot study with male participants suffering from burnout 
BMC Research Notes  2013;6:78.
Background
Occupational burnout is associated with severe negative health effects. While stress management programs proved to have a positive influence on the well-being of patients suffering from burnout, it remains unclear whether aerobic exercise alleviates burnout severity and other parameters related to occupational burnout. Therefore, the main purpose of this study was to pilot-test the potential outcomes of a 12-week exercise training to generate hypotheses for future larger scale studies.
Methods
The sample consisted of 12 male participants scoring high on the MBI emotional exhaustion and depersonalization subscales. The training program took place in a private fitness center with a 17.5 kcal/kg minimum requirement of weekly energy expenditure.
Results
The key findings are that increased exercise reduced overall perceived stress as well as symptoms of burnout and depression. The magnitude of the effects was large, revealing changes of substantial practical relevance. Additionally, profiles of mood states improved considerably after single exercise sessions with a marked shift towards an iceberg profile.
Conclusion
Among burnout patients, the findings provide preliminary evidence that exercise has the potential to reduce stress and prevent the development of a deeper depression. This has important health implications given that burnout is considered an antecedent of depressive disorders.
Trial registration
ClinicalTrials.gov Identifier: ISRNCT01575743
doi:10.1186/1756-0500-6-78
PMCID: PMC3599602  PMID: 23497731
Aerobic exercise; Burnout; Depressive symptoms; Maslach Burnout Inventory; Mood; Stress
12.  Evaluation of reference genes for reverse transcription quantitative PCR analyses of fish-pathogenic Francisella strains exposed to different growth conditions 
BMC Research Notes  2013;6:76.
Background
Reverse transcription quantitative PCR has become a powerful technique to monitor mRNA transcription in response to different environmental conditions in many bacterial species. However, correct evaluation of data requires accurate and reliable use of reference genes whose transcription does not change during the course of the experiment. In the present study exposure to different growth conditions was used to validate the transcription stability of eight reference gene candidates in three strains from two subspecies of Francisella noatunensis, a pathogen causing disease in both warm and cold water fish species.
Results
Relative transcription levels for genes encoding DNA gyrase (gyrA), RNA polymerase beta subunit (rpoB), DNA polymerase I (polA), cell division protein (ftsZ), outer membrane protein (fopA), riboflavin biosynthesis protein (ribC), 16S ribosomal RNA (16S rRNA) and DNA helicases (uvrD) were quantified under exponential, stationary and iron-restricted growth conditions. The suitability of selected reference genes for reliable interpretation of gene expression data was tested using the virulence-associated intracellular growth locus subunit C (iglC) gene.
Conclusion
Although the transcription stability of the reference genes was slightly different in the three strains studied, fopA, ftsZ and polA proved to be the most stable and suitable for normalization of gene transcription in Francisella noatunensis ssp.
doi:10.1186/1756-0500-6-76
PMCID: PMC3599356  PMID: 23452832
Gene expression; RT-qPCR; Francisella noatunensis; Fish; Francisellosis
13.  De novo transcriptomic resources for two sibling species of moths: Ostrinia nubilalis and O. scapulalis 
BMC Research Notes  2013;6:73.
Background
This study aimed at enhancing the transcriptomic resources for two sibling species of moths, Ostrinia scapulalis (Adzuki bean borer) and Ostrinia nubilalis (European corn borer), as a foundation for future researches on their divergence history. Previous works on these species had shown that their genetic divergence was low, while they were reproductively isolated in natura and specialized on different host plants. Comparative genomic resources will help facilitate the understanding of the mechanisms involved in this isolation and adaptation to the host plants. Despite their fundamental interest, these species still lack the genomic resources to thoroughly identify candidate genes for functions of interest. We present here a high throughput sequencing and de novo transcriptome assembly for these two sibling species in line with this objective of comparative genomics.
Results
Based on 322,504 and 307,622 reads of 454 sequencing for O. scapulalis and O. nubilalis respectively, we reconstructed 11,231 and 10,773 transcripts, of which 40% were functionally annotated by BLAST analyzes. We determined the level of completeness of both assemblies as well as the recovery level of published Ostrinia genomic resources. Gene ontology (GO) of common and species-specific de novo transcripts did not reveal GO terms significantly enriched in one or the other species. By applying stringent homology searches on transcripts common to O. scapulalis and O. nubilalis, we identified a set of homologous transcripts, with a mean nucleotide identity value of 98.1%. In this set, the most divergent transcripts revealed candidate genes involved in developmental, sensorial and pathogen defense processes.
Conclusions
This data greatly increases the genomic resources of Ostrinia species and constitute a solid skeleton for future comparative analyzes of expression or diversity, despite we show that the transcriptomes for both species have not been assembled at full completion. In addition, we provide a set of homologous transcripts together with their annotation as a source of candidate genes for comparative analyzes.
doi:10.1186/1756-0500-6-73
PMCID: PMC3599821  PMID: 23445568
NGS; 454 sequencing; Tanscriptome; Moth; European corn borer; Adzuki bean borer; Sibling species; Ostrinia nubilalis; Ostrinia scapulalis
14.  A preliminary study of individual cognitive behavior therapy for social anxiety disorder in Japanese clinical settings: a single-arm, uncontrolled trial 
BMC Research Notes  2013;6:74.
Background
Cognitive behavior therapy (CBT) is regarded as an effective treatment for social anxiety disorder (SAD) in Europe and North America. Individual CBT might be acceptable and effective for patients with SAD even in non-Western cultures; therefore, we conducted a feasibility study of individual CBT for SAD in Japanese clinical settings. We also examined the baseline predictors of outcomes associated with receiving CBT.
Methods
This single-arm trial employed a 14-week individual CBT intervention. The primary outcome was the self-rated Liebowitz Social Anxiety Scale, with secondary measurements of other social anxiety and depressive severity. Assessments were conducted at baseline, after a waiting period before CBT, during CBT, and after CBT.
Results
Of the 19 subjects screened, 15 were eligible for the study and completed the outcome measures at all assessment points. Receiving CBT led to significant improvements in primary and secondary SAD severity (ps < .001). The mean total score on the Liebowitz Social Anxiety Scale improved from 91.8 to 51.7 (before CBT to after CBT), and the within-group effect size at the end-point assessment was large (Cohen’s d = 1.71). After CBT, 73% of participants were judged to be treatment responders, and 40% met the criteria for remission. We found no significant baseline predictors of those outcomes.
Conclusion
Despite several limitations, our treatment—which comprises a 14-week, individual CBT program—seems feasible and may achieve favorable treatment outcomes for SAD in Japanese clinical settings. Further controlled trials are required in order to address the limitations of this study.
Trial registration
UMIN-CTR UMIN000005897
doi:10.1186/1756-0500-6-74
PMCID: PMC3602169  PMID: 23448435
Cognitive behavioral therapy; CBT; Social anxiety disorder; Social phobia; SAD; Japanese
15.  The illusion of specific capture: surface and solution studies of suboptimal oligonucleotide hybridization 
BMC Research Notes  2013;6:72.
Background
Hybridization based assays and capture systems depend on the specificity of hybridization between a probe and its intended target. A common guideline in the construction of DNA microarrays, for instance, is that avoiding complementary stretches of more than 15 nucleic acids in a 50 or 60-mer probe will eliminate sequence specific cross-hybridization reactions. Here we present a study of the behavior of partially matched oligonucleotide pairs with complementary stretches starting well below this threshold complementarity length – in silico, in solution, and at the microarray surface. The modeled behavior of pairs of oligonucleotide probes and their targets suggests that even a complementary stretch of sequence 12 nt in length would give rise to specific cross-hybridization. We designed a set of binding partners to a 50-mer oligonucleotide containing complementary stretches from 6 nt to 21 nt in length.
Results
Solution melting experiments demonstrate that stable partial duplexes can form when only 12 bp of complementary sequence are present; surface hybridization experiments confirm that a signal close in magnitude to full-strength signal can be obtained from hybridization of a 12 bp duplex within a 50mer oligonucleotide.
Conclusions
Microarray and other molecular capture strategies that rely on a 15 nt lower complementarity bound for eliminating specific cross-hybridization may not be sufficiently conservative.
doi:10.1186/1756-0500-6-72
PMCID: PMC3599332  PMID: 23445545
16.  A fast and efficient method for preparation of high-quality RNA from fungal mycelia 
BMC Research Notes  2013;6:71.
Background
Fungal RNA samples are usually isolated from fungal mycelia grown in liquid culture, which relies on prolific growth of the fungus in liquid media. The fungal biomass is then collected by vacuum filtration, which can result in low recovery for samples with reduced biomass due to poor growth in liquid media.
Findings
Here we report an alternative culturing method, based on growth on solid media which is independent of the ability of a fungus to grow in liquid culture. We show that growth on solid media overlayed with a nylon membrane is superior to other culturing methods, producing large amounts of biomass and allowing for easy harvesting of fungal mycelia. Furthermore, we show that mycelium harvested with this method yielded high-quality RNA, superior to RNA isolated from liquid grown mycelium. We also show that inclusion of a second chloroform extraction step in the procedure significantly increases RNA yield.
Conclusions
This method is particularly useful for fungal species that show poor or no growth in liquid media, but are easily cultured on solid media. Culturing can be performed on small petri dishes, which significantly reduces handling and therefore allowing growth and isolation of RNA from multiple strains in a high throughput manner. The obtained RNA samples are of high quality in sufficient quantities for several northern blot experiments or quantitative RT-PCR experiments.
doi:10.1186/1756-0500-6-71
PMCID: PMC3598987  PMID: 23442734
Fungi; RNA isolation; Fungal growth; Mycelium
17.  A preparation of murine liver fragments for in vitro studies: liver preparation for toxicological studies 
BMC Research Notes  2013;6:70.
Background
The aim of this study was to develop liver tissue preparation suitable for investigating toxins. Hepatocyte respiration, ATP content, urea synthesis, caspase activity and morphology were measured as a function of in vitro incubation time. Mice were anesthetized by sevoflurane inhalation. Small liver fragments were then rapidly excised and incubated at 37°C in Krebs-Henseleit buffer (continuously gassed with 95% O2: 5% CO2) for up to 6 h. Phosphorescence O2 analyzer was used to determine the rate of cellular mitochondrial O2 consumption (kc, μM O2 min-1 mg-1). Cellular ATP was measured using the luciferin/luciferase system. The caspase-3 substrate N-acetyl-asp-glu-val-asp-7-amino-4-methylcoumarin (Ac-DEVD-AMC) was used to monitor intracellular caspase activity; cleaved AMC moieties (reflecting caspase activity) were separated on HPLC and detected by fluorescence.
Findings
Respiration was inhibited by cyanide, confirming the oxidation occurred in the respiratory chain. The values of kc (mean ± SD) for 0≤ t ≤6 h were 0.15 ± 0.02 μM O2 min-1 mg-1 (n = 18, coefficient of variation, CV = 13%), ATP content 131 ± 69 pmol mg-1 (1≤ t ≤6 h, n = 16, CV = 53%), synthesized urea 0.134 ± 0.017 mg/dL mg-1 in 50 min (0≤ t ≤6 h, n = 14, CV = 13%), and AMC peak area 62,540 ± 26,227 arbitrary units mg-1 (1≤ t ≤6 h, n = 3, CV = 42%). Hepatocyte morphology and organelles were reasonably persevered.
Conclusions
The described liver tissue preparation demonstrates stable hepatocyte structure, ultrastructure and biomarkers for up to 6 h, permitting in vitro studies.
doi:10.1186/1756-0500-6-70
PMCID: PMC3598918  PMID: 23442607
In vitro; Cytotoxicity; Apoptosis; Liver; Mice; Cellular respiration; Bioenergetics; Caspases
18.  SHIFT: Server for hidden stops analysis in frame-shifted translation 
BMC Research Notes  2013;6:68.
Background
Frameshift is one of the three classes of recoding. Frame-shifts lead to waste of energy, resources and activity of the biosynthetic machinery. In addition, some peptides synthesized after frame-shifts are probably cytotoxic which serve as plausible cause for innumerable number of diseases and disorders such as muscular dystrophies, lysosomal storage disorders, and cancer. Hidden stop codons occur naturally in coding sequences among all organisms. These codons are associated with the early termination of translation for incorrect reading frame selection and help to reduce the metabolic cost related to the frameshift events. Researchers have identified several consequences of hidden stop codons and their association with myriad disorders. However the wealth of information available is speckled and not effortlessly acquiescent to data-mining. To reduce this gap, this work describes an algorithmic web based tool to study hidden stops in frameshifted translation for all the lineages through respective genetic code systems.
Findings
This paper describes SHIFT, an algorithmic web application tool that provides a user-friendly interface for identifying and analyzing hidden stops in frameshifted translation of genomic sequences for all available genetic code systems. We have calculated the correlation between codon usage frequencies and the plausible contribution of codons towards hidden stops in an off-frame context. Markovian chains of various order have been used to model hidden stops in frameshifted peptides and their evolutionary association with naturally occurring hidden stops. In order to obtain reliable and persuasive estimates for the naturally occurring and predicted hidden stops statistical measures have been implemented.
Conclusions
This paper presented SHIFT, an algorithmic tool that allows user-friendly exploration, analysis, and visualization of hidden stop codons in frameshifted translations. It is expected that this web based tool would serve as a useful complement for analyzing hidden stop codons in all available genetic code systems. SHIFT is freely available for academic and research purpose at http://www.nuccore.org/shift/.
doi:10.1186/1756-0500-6-68
PMCID: PMC3598200  PMID: 23432998
Frameshift; Reading frames; Hidden stop codons; Codon usage
19.  Is vancomycin MIC creep a worldwide phenomenon? Assessment of S. aureus vancomycin MIC in a tertiary university hospital 
BMC Research Notes  2013;6:65.
Background
Vancomycin is the primary treatment for infections caused by methicilin-resistant Staphylococcus aureus (MRSA). The association of vancomycin treatment failures with increased vancomycin minimum inhibitory concentration (MIC) is a well-recognized problem. A number of single-centre studies have identified progressive increases in glycopeptide MICs for S. aureus strains over recent years – a phenomenon known as vancomycin MIC creep. It is unknown if this is a worldwide phenomenon or if it is localized to specific centers.
Methods
The aim of this study was to evaluate the trend of vancomycin MIC for isolates of MRSA over a 3-year period in a tertiary university hospital in Portugal. MRSA isolates from samples of patients admitted from January 2007 to December 2009 were assessed. Etest method was used to determine the respective vancomycin MIC. Only one isolate per patient was included in the final analysis.
Results
A total of 93 MRSA isolates were studied. The vancomycin MICs were 0.75, 1, 1.5 and 2 mg/L for 1 (1.1%), 19 (20.4%), 38 (40.9%), 35 (37.6%) isolates, respectively. During the 3 year period, we observed a significant fluctuation in the rate of MRSA with a vancomycin MIC > 1 mg/L (2007: 86.2%; 2008: 93.3%; 2009: 58.8%, p = 0.002). No MRSA isolate presented a MIC > 2 mg/L.
Conclusions
We were unable to find in our institution data compatible to the presence of vancomycin MIC creep during the study period. This phenomenon seems not to be generalized; as a result each institution should systematically monitor MRSA vancomycin MIC over time.
doi:10.1186/1756-0500-6-65
PMCID: PMC3585458  PMID: 23422012
20.  Erosive potential of energy drinks on the dentine surface 
BMC Research Notes  2013;6:67.
Background
Considering the current high consumption of energy drinks, the aim of the present study is to evaluate the influence of energy drinks in removing the smear layer and exposing dentinal tubules on root surface.
Methods
Dentine root surfaces were exposed using a diamond bur. Forty movements of scaling were performed in the area prepared in order to create a smear layer. One hundred and thirty specimens were obtained from 35 teeth. Specimens were randomly distributed into 12 groups (n = 10) and divided into subgroups according to the application: topical (n = 5) and friction (n = 5). Twelve energy drinks were evaluated: RedBull™, Burn™, TNT™, Flash Power™, Flying Horse™, Sports Drink™, Ionic™, Hot Power™, Army Power™, Gladiator™ and Bug™. Distilled water was used as a control group. The specimens were analysed by scanning electron microscopy.
Results
Topical application: a significant influence of energy drinks on smear layer removal was found for FlyingHorse™ and Bug™ when compared with the control group. Friction application: significant smear layer removal was found for Burn™, FlyingHorse™, Gladiator™, SportsDrinks™, when compared with the control group. Comparing the different application forms, a statistically significant difference was found for Army Power™.
Conclusion
Considering the significant smear layer removal, energy drinks can be an important etiological factor for cervical dentine hypersensitivity.
doi:10.1186/1756-0500-6-67
PMCID: PMC3599422  PMID: 23422044
Dentine hypersensitivity; Erosive tooth wear; Scanning electron microscopy; Dentinal tubules; Abrasion; Tooth wear; Root dentin; Toothbrushing
21.  A novel long term short interval physical activity regime improves body composition in mice 
BMC Research Notes  2013;6:66.
Background
Exercise training (ET) and physical activity (PA) offer obvious health benefits in regular participants. In pre-clinical animal models, treadmills and running wheels are the models of choice for intervention studies using ET and PA. However, the frequency and duration necessary for positive effects on health are not completely understood. We investigated the impact of short duration voluntary wheel running on body composition in FVB × C57BL/6 F1 hybrid mice over 22 weeks. Mice were randomized and given access to voluntary wheel running (N = 6) or locked wheels (N = 5) for 1 hour per night during the dark cycle, 5 days per week.
Finding
Average weekly running distance was generally cyclic in nature over the 22 weeks but did not change significantly from week to week, except for a difference between week 3 and week 9 (P = 0.05). Daily running distances ranged from 0.78 km to 1.45 km. Compared with non-runners, runners demonstrated significantly lower relative fat mass (9.98 ± 0.56% vs. 14.91 ± 1.47%, P = 0.0067) and significantly higher relative lean mass (79.18 ± 0.65% vs. 75.41 ± 1.28%, P = 0.019). No differences were observed with respect to glucose metabolism.
Conclusion
Voluntary wheel running for one hour a day five days a week over a five month period improved body composition in young adult mice. This repetitive short interval exercise regime should be a useful model to investigate the effects of structured moderate intensity physical activity on physiological performance and chronic disease conditions in mice.
doi:10.1186/1756-0500-6-66
PMCID: PMC3599771  PMID: 23422015
Long term physical activity; Wheel running; FVB; C57BL/6; F1 hybrid mice; Fat mass; Lean body mass; Short interval running
22.  Y chromosome of the inbred mouse KK/Ta strain is associated with reduced body size in Y-consomic strains 
BMC Research Notes  2013;6:64.
Background
We have established 17 Y chromosome consomic (Y-consomic) mouse strains in an inbred DH/Sgn strain. In this study, based on investigations in four different genetic backgrounds, we proved that the Y chromosome of the inbred mouse KK/Ta strain is associated with reduced body size.
Findings
In the DH-Chr Y-+/+ background, Y chromosome substitution significantly decreased the body weight in DH-Chr YKK-+/+ and DH-Chr YSJL-+/+ strains, and the DH-Chr YKK-+/+ strain was the lightest among the 17 Y-consomic strains. In the DH-Chr Y-Dh/+ background (Dh/+ mice have skeletal malformations and are usually lighter than +/+ mice), although Y chromosome substitution did not significantly alter the body weight, the DH-Chr YKK-Dh/+ strain was the lightest among the 17 Y-consomic-Dh/+ strains. In the (B6.Cg-Ay × DH-Chr Y) F1-+/+ background, Y chromosome substitution significantly decreased the body weight and length in the (B6.Cg-Ay × DH-Chr YKK) F1 hybrids. In the (B6.Cg-Ay × DH-Chr Y) F1-Ay/+ background (Ay causes obesity and promotes linear growth), Y chromosome substitution significantly decreased body weight and length in the (B6.Cg-Ay × DH-Chr YKK) F1-Ay/+ hybrids.
Conclusion
A body-size-reducing effect of the Y chromosome of the KK/Ta mouse strain was observed irrespective of genetic background. The effect was observed in the presence of Dh and Ay, the autosomal dominant mutations, both of which are known to have substantial effects on body size. These results suggest that there are Y-linked genes that control the body size in mice.
doi:10.1186/1756-0500-6-64
PMCID: PMC3598876  PMID: 23418893
Ay allele; Body length; Body weight; Body size; Consomic mice; Dh; Y chromosome
23.  Maternal and fetal recovery after severe respiratory failure due to influenza: a case report 
BMC Research Notes  2013;6:62.
Background
During pregnancy women are at increased risk of severe complications to influenza infection, including death of mother or fetus, especially if chronic comorbid medical conditions such as diabetes mellitus are present.
Case presentation
A 36 years old Caucasian pregnant woman with type 1 diabetes underwent mechanical ventilation in gestation week 27 for severe respiratory failure due to influenza and pneumonia. For three weeks during and following her most severe illness, fetal growth could not be detected and the umbilical flows and amniotic fluid volumes were affected too. The possibility of preterm delivery and extracorporeal membrane oxygenation (ECMO) treatment were considered, however the patient and her fetus recovered gradually on conservative treatment. Under close surveillance the pregnancy continued until term, with delivery of an infant with appropriate weight for gestational age.
Conclusion
Preterm delivery and decreased birth weight were reported for women with antepartum pneumonia. Mechanical ventilation and ECMO treatment for severe respiratory failure in pregnancy are life threatening conditions and have been associated with preterm delivery. It remains uncertain if delivery improves the respiratory status of a critically ill woman, and the fetal condition is likely to improve, if the maternal condition is stabilized.
Severe respiratory insufficiency requiring mechanical ventilation in a diabetic pregnant woman with influenza was successfully treated conservatively. Despite clear signs of impaired fetal condition in the acute phase, watchful waiting resulted in delivery of a normal weight infant at term.
doi:10.1186/1756-0500-6-62
PMCID: PMC3583672  PMID: 23414816
Pregnancy; Diabetes; Influenza; Pneumonia; Mechanical ventilation; Pregnancy outcome
24.  Is estimated bullet trajectory a reliable predictor of severe injury? Case report of a thoraco-abdominal gunshot with a protracted trajectory managed nonoperatively 
BMC Research Notes  2013;6:63.
Background
Operative management of all gunshot’s traumas carries a high rate of unwarranted interventions that are known to cause serious complications. Selective nonoperative management is thus being increasingly practiced which has reduced these avoidable interventions. Physical examination and computed tomography scans are most sensitive in assessing need of laparotomy. Assessment of internal injuries on the basis of an estimated bullet trajectory is often practiced but has seldom been studied. We report a case of conservative management of a thoraco abdominal gun shot patient where an estimated bullet trajectory was indicative of serious injuries. To the best of our knowledge this is the first report of a thoraco abdominal gunshot that, despite of a protracted trajectory, had no sequelae and was thus managed nonoperatively.
Case presentation
A 30 year old male patient having height of 180 cm and weight of 70 kg (Body Mass Index 21.6) presented with complaint of a penetrating injury at left side of upper torso. The patient had no symptoms or obvious bleeding and was vitally stable. On examination a 1 cm × 1 cm entry wound at the left 3rd intercostal space in the mid clavicular line was identified. The chest and abdomen were otherwise unremarkable on examination. The chest radiograph displayed clear lung fields. The abdominal radiographs displayed a bullet in the upper left quadrant of the abdomen lateral to the spine. The bullets estimated trajectory from 3rd intercostal space and its lodgment in the abdomen lateral to the spine indicated severe visceral injury. The computed tomography scan showed that the bullet was lodged postero-medially to the left kidney. All thoracic, intra peritoneal and retroperitoneal visceral structures were identified to be normal. The patient remained clinically and vitally stable, hence was managed nonoperatively being discharged after 48 h of observation.
Conclusion
From this case we conclude that decision for managing gun shot patients should be based on objective clinical and diagnostic findings. We recommend further investigation of the predictability of estimated trajectory for visceral injuries and consequent operative intervention as we found it to be misleading in this case.
doi:10.1186/1756-0500-6-63
PMCID: PMC3598892  PMID: 23414905
25.  Endocrine and inflammatory profiles in type 2 diabetic patients with and without major depressive disorder 
BMC Research Notes  2013;6:61.
Background
There is a high prevalence of depression in individuals with type 2 diabetes mellitus. Depressive disorders are associated with increased medical morbidity and mortality in individuals with diabetes. It has been demonstrated that there is a higher prevalence of diabetic complications among individuals with diabetes and depression compared to those without depression. Several biological alterations have been reported in individuals with depressive disorders, particularly abnormal levels of endocrine-inflammatory markers.
This study aims to determine the prevalence of major depressive disorder (MDD) in type 2 diabetes patients, the prevalence of cardiovascular events in individuals with and without MDD and to compare the endocrine-inflammatory profile between groups.
Methods
The study was approved by the “Comité de Etica de Protocolos de Investigación del Departamento de Docencia e Investigación del Hospital Italiano de Buenos Aires” with the number “1262” and included only patients who provided written informed consent. The study was conducted in accordance with the Declaration of Helsinki and the Habeas Data law on protection of personal data (Law Nª 25326, Argentina).
Type 2 diabetes patients (n = 61) were included and they were classified as having MDD or not according to DSM-IV. Macrovascular disease was obtained from the medical history. Additionally, the intima-media thickness of the common carotid, carotid bifurcations and internal carotid arteries was measured non-invasively by two-dimensional ultrasound imaging. Fasting glucose, fasting lipid profile, inflammatory (CRP, TNF-α) and endocrine (urine free cortisol and saliva cortisol) markers. Student t tests were used to compare means for normally distributed variables and Mann-Whitney test for variables without normal distribution. Relative frequencies were calculated and a chi-square analysis was conducted. Data were expressed as mean ± standard deviation (SD) or median and interquartile range. Multivariable logistic regression was used to determine the relative odds of clinical cardiovascular disease in individuals with compared to those without depression. Differences were considered significant using a two-sided p < 0.05.
Results
21 patients (34%) had MDD and 40 patients (66%) didn’t have MDD. Diabetic patients with MDD had significantly higher CRP levels (4.1(1.9-7.6) vs 1.5(0.5-4.4) mg/l; p = 0.02) and 24-hour urine free cortisol (71.4 ± 21.3 vs 59.8 ± 29.3 ug/24 h; p = 0.03). The other metabolic and inflammatory parameters were not statistically different between groups. There was a significantly higher prevalence of cardiovascular events in individuals with MDD: 38% for the depressive group vs 15% for non-depressive group, p = 0.04). Patients with MDD had a 3.5-fold greater odd of having cardiovascular disease.
Conclusions
Diabetic patients with depression are more likely to have cardiovascular events, and different factors can determine this high association.
doi:10.1186/1756-0500-6-61
PMCID: PMC3599430  PMID: 23410093
Depression; Type 2 diabetes mellitus; Cardiovascular disease

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