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1.  Infection of human monocytes by Chlamydia pneumoniae and Chlamydia trachomatis: an in vitro comparative study 
BMC Research Notes  2014;7:230.
An increasing number of studies suggest that chlamydiae can infect immune cells. The altered immune cell function could contribute to the progression of several chronic inflammatory diseases.
The aim of this study was to comparatively evaluate Chlamydia pneumoniae (CP) and Chlamydia trachomatis (CT) interactions with in vitro infected human blood monocytes.
Fresh isolated monocytes were infected with viable CP and CT elementary bodies and infectivity was evaluated by recultivating disrupted monocytes in permissive epithelial cells.
The production of reactive oxygen and nitrogen species was studied in the presence of specific fluorescent probes. Moreover, TNF-α, INF-α, INF-β and INF-γ gene expression was determined.
CT clearance from monocytes was complete at any time points after infection, while CP was able to survive up to 48 hours after infection. When NADPH oxydase or nitric oxide synthase inhibitors were used, CT infectivity in monocytes was restored, even if at low level, and CT recovery’s rate was comparable to CP one.
CT-infected monocytes produced significantly higher levels of reactive species compared with CP-infected monocytes, at very early time points after infection. In the same meanwhile, TNF-α and INF-γ gene expression was significantly increased in CT-infected monocytes.
Our data confirm that CP, but not CT, is able to survive in infected monocytes up to 48 hours post-infection. The delay in reactive species and cytokines production by CP-infected monocytes seems to be crucial for CP survival.
PMCID: PMC3984436  PMID: 24721461
Human monocytes; Chlamydia pneumoniae; Chlamydia trachomatis; Reactive oxygen species; Reactive nitrogen species; Cytokines
2.  Translation and validation of European organization for research and treatment of cancer quality of life Questionnaire -C30 into Moroccan version for cancer patients in Morocco 
BMC Research Notes  2014;7:228.
Understanding the effects of cancer on the quality of life of affected patients is critical to clinical research as well as to optimal management and care. The aim of this study was to adapt the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 (EORTC QLQ-C30) questionnaire into Moroccan Arabic and to determine its psychometric properties. After translation, back translation and pretesting of the pre-final version, the translated version was submitted to a committee of professionals composed by oncologists and epidemiologists. The psychometric properties were tested in patients with cancer. Internal consistency was tested using Cronbach’s alpha and the test-retest reliability using interclass correlation coefficients. Construct validity was assessed by examining item-convergent and divergent validity. It was also tested using Spearman’s correlation between QLQ-C30 scales and EQ-5D.
The study was conducted in 125 patients. The Moroccan version was internally reliable, Cronbach’s α was 0.87 for the total scale and ranged from 0.34 to 0.97 for the subscales. The intraclass correlation coefficient of the test-retest reliability ranged from 0.64 for “social functioning” to 0.89 for “physical activities” subscales. The instrument demonstrated a good construct and concomitant validity.
We have developed a semantically equivalent translation with cultural adaptation of EORTC QLQ-C30 questionnaire. The assessment of its measurement properties showed that it is quite reliable and a valid measure of the effect of cancer on the quality of life in Moroccan patients.
PMCID: PMC3984435  PMID: 24721384
Cancer; Quality of life; Reliability; Validity
3.  Cardioprotective role of leaves extracts of Carissa opaca against CCl4 induced toxicity in rats 
BMC Research Notes  2014;7:224.
Carissa opaca are used traditionally in Pakistan for the treatment of various human ailments. Therefore, the study is arranged out to assess the cardio protective potential of different fractions of Carissa opaca leaves on CCl4-induced oxidative trauma in kidney.
The parameters studied in this respect were the cardiac function test (CK (U/l), CKMB (U/l), genotoxicity (% DNA fragmentation), characteristic morphological findings and antioxidant enzymatic level of cardiac tissue homogenate.
The protective effects of various fractions of Carissa opaca (C. opaca) leaves extract against CCl4 administration was reviewed by rat cardiac functions alterations. Chronic toxicity caused by eight week treatment of CCl4 to the rats significantly changed the cardiac function test, decreased the activities of antioxidant enzymes and glutathione contents whereas significant increase was found in lipid peroxidation comparative to control group. Administration of various fractions of C. opaca leaves extract with CCl4 showed protective ability against CCl4 intoxication by restoring the cardiac functions alterations, activities of antioxidant enzymes and lipid peroxidation in rat. CCl4 induction in rats also caused DNA fragmentation and histopathalogical abnormalities which were restored by co-admistration of various fraction of C. opaca leaves extract.
Results revealed that various fraction of C. opaca are helpful in cardiac dysfunctions.
PMCID: PMC3983670  PMID: 24716654
DNA damages; Lipid peroxidation; Histopathalogical; CCl4
4.  Identification and enzymatic characterization of acid phosphatase from Burkholderia gladioli 
BMC Research Notes  2014;7:221.
The genus Burkholderia is widespread in diverse ecological niches, the majority of known species are soil bacteria that exhibit different types of non-pathogenic interactions with plants. Burkholderia species are versatile organisms that solubilize insoluble minerals through the production of organic acids, which increase the availability of nutrients for the plant. Therefore these bacteria are promising candidates for biotechnological applications.
Burkholderia sp. (R 3.25 isolate) was isolated from agricultural soil in Ponta Grossa-PR-Brazil and identified through analysis of the 16S rDNA as a strain classified as Burkholderia gladioli. The expression of membrane-bound acid phosphatase (MBAcP) was strictly regulated with optimal expression at a concentration of phosphorus 5 mM. The apparent optimum pH for the hydrolysis of p-nitrophenylphosphate (PNPP) was 6.0. The hydrolysis of PNPP by the enzyme exhibited a hyperbolic relationship with increasing concentration of substrate and no inhibition by excess of substrate was observed. Kinetic data revealed that the hydrolysis of PNPP exhibited cooperative kinetics with n = 1.3, Vm = 113.5 U/mg and K0.5 = 65 μM. The PNPPase activity was inhibited by vanadate, p-hydroxymercuribenzoate, arsenate and phosphate, however the activity was not inhibited by calcium, levamisole, sodium tartrate, EDTA, zinc, magnesium, cobalt, ouabain, oligomycin or pantoprazol.
The synthesis of membrane-bound non-specific acid phosphatase, strictly regulated by phosphate, and its properties suggest that this bacterium has a potential biotechnological application to solubilize phosphate in soils with low levels of this element, for specific crops.
PMCID: PMC3983855  PMID: 24713147
Phosphohydrolase; Inhibition; Phosphate; P-Nitrophenylphosphate; Solubilization
5.  Musculoskeletal pain and school bag use: a cross-sectional study among Ugandan pupils 
BMC Research Notes  2014;7:222.
Though seen as a convenient method of carrying books and other scholastic materials including food items, schoolbags are believed to contribute to back and other musculoskeletal problems in school going children. This study set out to determine the prevalence of low back and other musculoskeletal pains and describe their relationship with schoolbag use in pupils.
This was a cross-sectional descriptive study involving 532 pupils from six primary schools with a mean age of 13.6 years. Analyses included the chi- square test, independent t tests, regression analysis and test for trend across ordered groups.
Backpacks were the most common type of schoolbag and younger children carried disproportionately heavier bags. Urban pupils were younger, carried significantly heavier bags, and less likely to complain about schoolbag weight than the rural pupils,
About 30.8% of the pupils carried schoolbags which were more than 10% of their body weight. About 88.2% of pupils reported having body pain especially in the neck, shoulders and upper back. About 35.4% of the children reported that carrying the schoolbag was the cause of their musculoskeletal pain. The prevalence of lower back pain was 37.8%. There was significant association between low back pain and; method of bag carriage (p < 0.0001), long duration of walking (odds ratio 2.67, 95% CI 1.38- 5.16) and the time spent sitting after school (p = 0.02). Only 19% had lockers at school.
Urban pupils were younger, carried significantly heavier bags, and less likely to complain about schoolbag weight than the rural pupils. The majority of pupils complained of musculoskeletal pain of which 35.4% was attributed to the schoolbags.
The prevalence of lower back pain was 37.8%. Schools need to provide lockers and functional libraries in order to avoid excessive loading and repetitive strain injuries.
PMCID: PMC3983861  PMID: 24713177
Musculoskeletal pain; Low back pain; Schoolbag weight; Pupils
6.  Prevalence and predictors of Lymphogranuloma venereum in a high risk population attending a STD outpatients clinic in Italy 
BMC Research Notes  2014;7:225.
We evaluated LGV prevalence and predictors in a high risk population attending a STI Outpatients Clinic in the North of Italy.
A total of 108 patients (99 MSM and 9 women), with a history of unsafe anal sexual intercourses, were enrolled. Anorectal swabs and urine samples were tested for Chlamydia trachomatis (CT) DNA detection by Versant CT/GC DNA 1.0 Assay (Siemens Healthcare Diagnostics Terrytown, USA). RFLP analysis was used for CT molecular typing.
L2 CT genotype was identified in 13/108 (12%) rectal swabs. All LGV cases were from MSM, declaring high-risk sexual behaviour and complaining anorectal symptoms. Patients first attending the STI Outpatient Clinic received a significant earlier LGV diagnosis than those first seeking care from general practitioners or gastroenterologists (P = 0.0046).
LGV prevalence and characteristics found in our population are in agreement with international reports. Statistical analysis showed that LGV positive patients were older (P = 0.0008) and presented more STIs (P = 0.0023) than LGV negative ones, in particular due to syphilis (P < 0.001), HIV (P < 0.001) and HBV (P = 0.001).
Multivariate logistic regression analysis revealed that HIV and syphilis infections are strong risk factors for LGV presence (respectively, P = 0.001 and P = 0.010).
Even if our results do not provide sufficient evidence to recommend routine screening of anorectal swabs in high-risk population, they strongly suggest to perform CT NAAT tests and genotyping on rectal specimens in presence of ulcerative proctitis in HIV and/or syphilis-positive MSM. In this context, CT DNA detection by Versant CT/GC DNA 1.0 Assay, followed by RFLP analysis for molecular typing demonstrated to be an excellent diagnostic algorithm for LGV identification.
PMCID: PMC3984434  PMID: 24716676
LGV; MSM; Chlamydia trachomatis; NAATs
7.  Telephone interviews and online questionnaires can be used to improve neurodevelopmental follow-up rates 
BMC Research Notes  2014;7:219.
Maximising response rates to neurodevelopmental follow-up is a key challenge for paediatric researchers. We have investigated the use of telephone interviews and online questionnaires to improve response rates, reduce non-response bias, maintain data completeness and produce unbiased outcomes compared with postal questionnaires when assessing neurodevelopmental outcomes at 2 years.
A prospective cohort study of babies born ≥32 weeks gestation. Neurodevelopmental outcomes were assessed at 2 years of age using a parent questionnaire completed via post, telephone or online. Relative Risks with 95% confidence intervals (RR; 95% CI) were calculated to identify participant characteristics associated with non-response and questionnaire response mode (postal vs. telephone/online). The proportion of missing data and prevalence of adverse outcomes was compared between response modes using generalized linear models.
Offering telephone/online questionnaires increased the study response rate from 55% to 60%. Telephone/online responders were more likely to be non-white (RR 1.6; [95% CI 1.1, 2.4]), non-English speaking (1.6; [1.0, 2.6]) or have a multiple birth (1.6; [1.1, 2.3]) than postal responders. There were no significant differences in the prevalence of adverse neurodevelopmental outcomes between those who responded via post vs. telephone/online (1.1; [0.9, 1.4]). Where parents attempted all questionnaire sections, there were no significant differences in the proportion of missing data between response modes.
Where there is sufficient technology and resources, offering telephone interviews and online questionnaires can enhance response rates and improve sample representation to neurodevelopmental follow-up, whilst maintaining data completeness and unbiased outcomes.
PMCID: PMC3983863  PMID: 24716630
Neurodevelopmental outcomes; Follow-up; Response rates; Cohort study; Questionnaire
8.  Virulence factors, serogroups and antimicrobial resistance properties of Escherichia coli strains in fermented dairy products 
BMC Research Notes  2014;7:217.
From a clinical perspective, it is essential to know the microbial safety of fermented dairy products. Doogh and kashk are fermented dairies. These products are used by millions of people but their microbial qualities are unknown. Shiga toxin producing Escherichia coli (STEC) is one of the most commonly detected pathogens in the cases of food poisoning and food-borne illnesses. The present investigation was carried out in order to study the molecular characterization and antimicrobial resistance properties of STEC strains isolated from fermented dairy products.
Six hundred fermented dairy samples were collected and immediately transferred to the laboratory. All samples were cultured immediately and those that were E. coli-positive were analyzed for the presence of O157 , O26, O103, O111, O145, O45, O91, O113, O121 and O128 STEC serogroups, tetA, tetB, blaSHV, CITM, cmlA, cat1, aadA1, dfrA1, qnr, aac (3)-IV, sul1 and ereA antibiotic resistance genes and stx1, stx2, eaeA, ehly, cnf1, cnf2, iutA, cdtB, papA, traT, sfaS and fyuA virulence factors using PCR. Antimicrobial susceptibility testing was performed also using disk diffusion methodology with Mueller–Hinton agar.
Fifty out of 600 (8.33%) dairy samples harbored E. coli. In addition, yoghurt was the most commonly contaminated dairy. O157 (26%) and O26 (12%) were the most commonly detected serogroups. A significant difference was found between the frequency of Attaching and Effacing E. coli and Enterohaemorrhagic E. coli (P <0.05). Stx1 (44%), eae (36%), papA (32%) stx2 (30%), and ehly (28%) were the most commonly detected virulence factors. The genes encode resistance against tetracycline (tetA and tetB) (76% and 70%, respectively), cephalothin (blaSHV) (38%), ampicillin (CITM) (36%) and gentamicin (aac (3)-IV) (32%) were the most commonly detected. High resistance levels to tetracycline (84%), penicillin (46%), ampicillin (38%) and streptomycin (36%) were observed.
Fermented dairy products can easily become contaminated by antibiotic resistant STEC strains. Our findings should raise awareness about antibiotic resistance in Iran. Clinicians should exercise caution when prescribing antibiotics, especially in veterinary treatments.
PMCID: PMC3983858  PMID: 24708594
Shiga toxin-producing Escherichia coli; Virulence factors; Antibiotic resistance properties; Fermented dairy products; Iran
9.  Association of adiponectin with hepatic steatosis: a study of 1,349 subjects in a random population sample 
BMC Research Notes  2014;7:207.
Objective of the present study was to examine the association between adiponectin and hepatic steatosis, and other biochemical and anthropometric parameters in healthy subjects.
A total of 1349 subjects (age 18–65 years) underwent ultrasound examination of the liver. Mean adiponectin concentration for the study collective was 11.35 ± 6.28 μg/mL. The following parameters were assessed for their association with adiponectin: body-mass index (BMI); age; sex; arterial blood pressure; nicotine use; alcohol consumption; physical activity; metabolic syndrome; total, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol; triglycerides; aspartate aminotransferase (AST); alanine aminotransferase (ALT); γ-glutamyltransferase (GGT); alkaline phosphatase (AP); C-reactive protein (CRP); insulin sensitivity according to the Homeostasis Model Assessment (HOMA); random blood glucose; and the degree of steatosis of the liver. The numerical differences in the variables influencing adiponectin returned in the descriptive analysis were confirmed at bivariate analysis for BMI, ALT, AST, GGT, AP, total and HDL cholesterol, triglycerides, CRP, arterial blood pressure, metabolic syndrome, nicotine use and alcohol consumption. The logistic regression of the multivariate analysis showed that male sex, hepatic steatosis, BMI, metabolic syndrome, tobacco smoking and CRP correlate negatively with adiponectin, while age, moderate alcohol consumption and HDL cholesterol exhibit a positive association.
The results of the present study confirm the findings of previous research. Adiponectin correlates negatively with cardiometabolic risk factors and is an independent indicator for non-alcoholic fatty liver disease (NAFLD).
PMCID: PMC3977975  PMID: 24693952
Ultrasonography; Fatty liver; NAFLD; Adipose tissue; Cross-sectional studies
11.  Feasibility of self-collection of fecal specimens by randomly sampled women for health-related studies of the gut microbiome 
BMC Research Notes  2014;7:204.
The field of microbiome research is growing rapidly. We developed a method for self-collection of fecal specimens that can be used in population-based studies of the gut microbiome. We conducted a pilot study to test the feasibility of our methods among a random sample of healthy, postmenopausal women who are members of Kaiser Permanente Colorado (KPCO). We aimed to collect questionnaire data, fecal and urine specimens from 60 women, aged 55–69, who recently had a normal screening mammogram. We designed the study such that all questionnaire data and specimens could be collected at home.
We mailed an invitation packet, consent form and opt-out postcard to 300 women, then recruited by telephone women who did not opt-out. Verbally consented women were mailed an enrollment package including a risk factor questionnaire, link to an online diet questionnaire, specimen collection kit, and instructions for collecting stool and urine. Specimens were shipped overnight to the biorepository. Of the 300 women mailed an invitation packet, 58 (19%) returned the opt-out postcard. Up to 3 attempts were made to telephone the remaining women, of whom 130 (43%) could not be contacted, 23 (8%) refused, and 12 (4%) were ineligible. Enrollment packages were mailed to 77 women, of whom 59 returned the risk factor questionnaire and specimens. We found no statistically significant differences between enrolled women and those who refused participation or could not be contacted.
We demonstrated that a representative sample of women can be successfully recruited for a gut microbiome study; however, significant personal contact and carefully timed follow-up from the study personnel are required. The methods employed by our study could successfully be applied to analytic studies of a wide range of clinical conditions that have been postulated to be influenced by the gut microbial population.
PMCID: PMC3974920  PMID: 24690120
Study design; Microbiome; Breast cancer
12.  Pattern of neurological disease seen among patients admitted in tertiary care hospital 
BMC Research Notes  2014;7:202.
Neurologic disorders are not uncommon at in patient departments of different hospitals. We have conducted the study to see the pattern and burden of neurologic disorders at different inpatient departments of a tertiary care centre.
This retrospective observational study was carried out from the records and referral notes of neurology department of Dhaka Medical College Hospital (DMCH) from July 2011 to June 2012. A total 335 patients were evaluated by consultant neurologists during this period.
Majority of the patients (59.7%) presented after the age of forty years. The mean age at presentation was 45.11 ± 17.3 years with a male predominance (63.3%). Stroke was the most common condition (47.5%) observed at referral, followed by seizure (9.3%), disease of spinal cord (7.8%) and encephalopathy (6.3%). Even after consultation, 30 patients remained undiagnosed and 6 were diagnosed as functional disorder. Department of Medicine (231, 69%) and Cardiology (61, 18.2%) made most of the calls. More than half (56%) of the stroke patients were referred from medicine and one third (35.2%) from cardiology. Seizure (67.7%), problem in spinal cord (92.3%), coma (50%), encephalopathy (57.1%), motor neuron disease (MND) (72.7%) were common reasons for referral from department of Medicine. Whereas patients with cord disease (7.3%), CNS tumor (40%), seizure disorder (6.5%) and stroke (3.8%) were referred from surgery. Department of Obstetrics and Gynecology sought help for stroke (2.5%), seizure (12.9%), MND (27.3%), coma (16.7%) and encephalopathy (9.5%).
Hypertension, diabetes, ischemic heart disease, dyslipidaemia and respiratory problem were significantly associated co-morbid conditions in stroke patients (at 95% CI, p value is <0.001, <0.01, <0.001, <0.05, <0.05 respectively). Hematological disorders were common association among patients with cord problem (<0.05).
Wide ranges of neurological problems are often managed by physicians and surgeons, especially those from medicine and cardiology. Where ever available consultation from neurologists can help in diagnosing and managing these cases.
PMCID: PMC3977680  PMID: 24684800
Neurologic disease
13.  Novel missense mutation in the FH gene in familial renal cell cancer patients lacking cutaneous leiomyomas 
BMC Research Notes  2014;7:203.
Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a rare tumor predisposition syndrome characterized by cutaneous and uterine leiomyomas and papillary type 2 renal cell cancer. Germline mutation of the fumarate hydratase (FH) gene is known to be associated with HLRCC.
Case presentation
We describe a 64-year-old father and his 39-year-old son with HLRCC who developed papillary type 2 RCCs lacking cutaneous leiomyomas at any site. A common missense mutation in the FH gene, (c.1021G > A, p.D341N) in exon 7, was detected in the 2 cases. Functional prediction with the bioinformatics programs, SIFT and Polyphen-2, reported “damaging (SIFT score 0.00)” and “probably damaging (PSIC score 1.621)” values, respectively. In 162 healthy individuals, there were no cases of a G transition to any base. Finally, (c.1021G > A) in exon 7, was identified as a point mutation.
We report a family with HLRCC in which a novel missense mutation was detected. A familial papillary type 2 renal cancer should be considered HLRCC unless typical cutaneous leiomyomas do not occur.
PMCID: PMC3978052  PMID: 24684806
Familial renal cell cancer; Papillary renal cell cancer; Fumarate hydratase; Missense mutation
14.  A pharmacist-led follow-up program for patients with coronary heart disease in North Norway–a qualitative study exploring patient experiences 
BMC Research Notes  2014;7:197.
Coronary heart disease (CHD) is one of the leading causes of death worldwide. Scientific literature shows that prevention of CHD is inadequate. The clinical pharmacist’s role in patient-centred care has been shown favourable in a large amount of studies, also in relation to reduction of risk factors related to CHD. We developed and piloted a pharmacist-led follow-up program for patients with established CHD after hospital discharge from a hospital in North Norway. The aim of the present study was to explore how participants in the follow-up program experienced the program with regard to four main topics; medication knowledge, feeling of safety and comfort with medications, the functionality of the program and the clinical pharmacist’s role in the interdisciplinary team.
We performed semi-structured thematic interviews with four patients included in the program. After verbatim transcribing, we analysed the interviews using “qualitative content analyses” by Graneheim and Lundman. Trial registration NCT01131715.
All participants appreciated the follow-up program because their medication knowledge had increased, participation had made them feel safe, they were reassured about the appropriateness of their medications, and they had become more involved in their own medication. The participants reported that the program was well structured and the clinical pharmacist was said to be an important caretaker in the health-care system. The importance of collaboration between pharmacists and physicians, both in hospital and primary care, was emphasized.
Our results indicate that the follow-up program was highly appreciated among the four participants included in this study. The results must be interpreted in the context of the health care system in Norway today. Here, few pharmacists are working in hospitals or in close relation to the general practitioners. In addition, physicians are short of time in order to supply appropriate medication information, both in hospital and primary care. Involving pharmacists in follow-up of patients with CHD seems to be highly appreciated among patients and may be a step towards improving patient care. The study is limited by the low number of participants.
PMCID: PMC3974183  PMID: 24679131
Qualitative study; Patient experience; Follow-up program; Clinical pharmacist; Coronary heart disease
15.  Pattern and factors associated with congenital anomalies among young infants admitted at Bugando medical centre, Mwanza, Tanzania 
BMC Research Notes  2014;7:195.
Congenital anomalies or birth defects are among the leading causes of infant mortality and morbidity around the world. The impact of congenital anomalies is particularly severe in middle- and low-income countries where health care resources are limited. The prevalence of congenital anomalies varies in different parts of the world, which could reflect different aetiological factors in different geographical regions.
Between October 2012 and January 2013, a cross-sectional study was conducted involving young infants below 2 months of age, admitted at a university teaching hospital in Tanzania. Face-to-face interviews with parents/caretakers of young infants were carried out to collect socio-demographic and clinical information. Physical examinations were performed on all young infants. Echocardiography, X-ray, cranial as well as abdominal ultrasonographies were performed when indicated.
Analysis of the data showed that among 445 young infants enrolled in the study, the prevalence of congenital anomalies was 29%, with the Central Nervous System (CNS) as the most commonly affected organ system. Maternal factors that were significantly associated with congenital anomalies included the lack of peri-conceptional use of folic acid (OR = 3.1; 95% CI = 1.4-6.7; p = 0.005), a maternal age of above 35 years (OR = 2.2; 95% CI = 1.1-4.3; p = 0.024) and an inadequate attendance to antenatal clinic (OR = 2.1; 95% CI = 1.4-3.3; p < 0.001). Infant factors that were significantly associated with congenital anomalies were female sex, a birth weight of 2.5 kg or more, singleton pregnancy and a birth order above 4.
Due to the high prevalence of congenital anomalies observed in this particular context, the hospital should mobilize additional resources for an optimal and timely management of the patients with congenital anomalies. In this study, the proportion of women taking folic acid supplements during early pregnancy was very low. Efforts should be made to ensure that more women use folic acid during the peri-conceptional period, as the use of folic acid supplement has been linked by several authors to a reduced occurrence of some congenital anomalies.
PMCID: PMC3974194  PMID: 24679067
16.  Distinct polymicrobial populations in a chronic foot ulcer with implications for diagnostics and anti-infective therapy 
BMC Research Notes  2014;7:196.
Polymicrobial infections caused by combinations of different bacteria are being detected with an increasing frequency. The evidence of such complex infections is being revealed through the use of novel molecular and culture-independent methods. Considerable progress has been made in the last decade regarding the diagnostic application of such molecular techniques. In particular, 16S rDNA-based sequencing and even metagenomic analyses have been successfully used to study the microbial diversity in ecosystems and human microbiota. Here, we utilized denaturing high-performance liquid chromatography (DHPLC) as a diagnostic tool for identifying different bacterial species in complex clinical samples of a patient with a chronic foot ulcer.
Case presentation
A 45-year-old female suffered from a chronic 5x5cm large plantar ulcer located in the posterior calcaneal area with subcutaneous tissue infection and osteomyelitis. The chronic ulcer developed over a period of 8 years. Culture and DHPLC revealed a distinct and location-dependent polymicrobial infection of the ulcer. The analysis of a superficial biopsy revealed a mixture of Staphylococcus aureus, Proteus vulgaris, and Fusobacterium nucleatum, whereas the tissue-deep biopsy harbored a mixture of four different bacterial species, namely Gemella morbillorum, Porphyromonas asaccharolytica, Bacteroides fragilis, and Arcanobacterium haemolyticum.
This clinical case highlights the difficulties in assessing polymicrobial infections where a mixture of fastidious, rapid and slow growing bacteria as well as anaerobes exists as structured communities within the tissue architecture of chronic wound infections. The diagnosis of this multilayered polymicrobial infection led to a microbe-adapted antibiotic therapy, targeting the polymicrobial nature of this infection in addition to a standard local wound treatment. However, a complete wound closure could not be achieved due to the long-lasting extensive destruction of tissue.
PMCID: PMC3974921  PMID: 24679105
Polymicrobial infection; Foot ulcer; Microbiome; Gemella morbillorum
17.  Inferior vena cava tumor thrombus after partial nephrectomy for renal cell carcinoma 
BMC Research Notes  2014;7:198.
Partial nephrectomy is now the gold standard treatment for small renal tumors. Local recurrence is a major problem after partial nephrectomy, and local recurrence in the remnant kidney after partial nephrectomy is common.
Case presentation
A 77-year-old man underwent right partial nephrectomy for a T1 right renal cell carcinoma. Microscopic examination revealed a clear cell renal carcinoma, grade 2, stage pT3a. Although the surgical margin was negative, the carcinoma invaded the perirenal fat, and vascular involvement was strongly positive. Thirty months after partial nephrectomy, an enhanced computed tomographic scan showed local recurrence of the renal cell carcinoma extending into the inferior vena cava without renal mass. Hence, we performed right radical nephrectomy and intracaval thrombectomy. Microscopic examination revealed a clear cell carcinoma grade 2, stage pT3a + b. The patient is still alive with no evidence of recurrence 10 months post-procedure.
To our knowledge, local recurrence of renal cell carcinoma extending into the inferior vena cava after partial nephrectomy has not been reported in the literature. Our case report emphasizes the importance of strict surveillance of patients after partial nephrectomy, especially for those with renal cell carcinoma positive for microvessel involvement.
PMCID: PMC3977664  PMID: 24679158
Partial nephrectomy; Renal cell carcinoma; Local recurrence; Tumor thrombus
18.  Exploring relationships between whole carcass condemnation abattoir data, non-disease factors and disease outbreaks in swine herds in Ontario (2001–2007) 
BMC Research Notes  2014;7:185.
Improving upon traditional animal disease surveillance systems may allow more rapid detection of disease outbreaks in animal populations. In Ontario, between the years 2001 – 2007, widespread outbreaks of several diseases caused major impacts to the swine industry. This study was undertaken to investigate whether whole carcass condemnation data of market pigs from provincial abattoirs from 2001 – 2007 could have provided useful information for disease surveillance of Ontario swine. The objective was to examine the suitability of these data for detection of disease outbreaks using multi-level models and spatial scan statistics. We investigated the ability of these data to provide spatially-relevant surveillance information by determining the approximate distance pigs are shipped from farm to provincial abattoirs in the province, and explored potentially biasing non-disease factors within these data.
Provincially-inspected abattoirs in Ontario were found to be located in close proximity to the hog farms of origin. The fall season and increasing abattoir capacity were associated with a decrease in condemnation rates. Condemnation rates varied across agricultural regions by year, and some regions showed yearly trends consistent with the timing of emergence of new disease strains that affected the Ontario swine population. Scan statistics identified stable clusters of condemnations in space that may have represented stable underlying factors influencing condemnations. The temporal scans detected the most likely cluster of high condemnations during the timeframe in which widespread disease events were documented. One space-time cluster took place during the beginning of the historical disease outbreaks and may have provided an early warning signal within a syndromic surveillance system.
Spatial disease surveillance methods may be applicable to whole carcass condemnation data collected at provincially-inspected abattoirs in Ontario for disease detection on a local scale. These data could provide useful information within a syndromic disease surveillance system for protecting swine herd health within the province. However, non-disease factors including region, season and abattoir size need to be considered when applying quantitative methods to abattoir data for disease surveillance.
PMCID: PMC3975458  PMID: 24674622
19.  Septo-optic dysplasia plus: a case report 
BMC Research Notes  2014;7:191.
Septo-optic dysplasia, also referred to as de Morsier syndrome, is a congenital condition characterized by classic triad features: midline brain abnormalities, optic nerve hypoplasia and pituitary endocrine dysfunction. Sometimes, other various malformations appear within syndrome.
Case presentation
An 11 and 1/2-year-old Caucasian Southeast European female patient with earlier established diagnoses of growth hormone deficiency, diabetes insipidus, seizures, mental retardation, optic nerve atrophy and right ptosis, was directed to us for consultative examination.
The girl of short stature and low weight for her age had bilateral optic nerve hypoplasia, poor vision, nystagmus and right eye oculomotor palsy. Electroencephalogram revealed epileptic changes. Magnetic resonance imaging showed an empty sella syndrome, partial hypoplasia of corpus callosum, cavum of pellucid septum and diffuse polymicrogyria of the left temporal lobe. We found all elements of septo-optic dysplasia plus syndrome with right oculomotor nerve involvement.
By earlier findings and evaluation, we established a diagnosis of septo-optic dysplasia plus. The case confirms the existence of various malformations within the syndrome and the need for the cooperation of several specialists in the diagnosis and treatment of children with the syndrome.
PMCID: PMC3976455  PMID: 24678945
Septo-optic dysplasia; Optic hypoplasia; Polymicrogyria; Hypopituitarism; Oculomotor palsy
20.  Disruption of mutated BRAF signaling modulates thyroid cancer phenotype 
BMC Research Notes  2014;7:187.
Thyroid cancer is the most common endocrine-related cancer in the United States and its incidence is rising rapidly. Since among various genetic lesions identified in thyroid cancer, the BRAFV600E mutation is found in 50% of papillary thyroid cancers and 25% of anaplastic thyroid cancers, this mutation provides an opportunity for targeted drug therapy. Our laboratory evaluated cellular phenotypic effects in response to treatment with PLX4032, a BRAFV600E-specific inhibitor, in normal BRAF-wild-type thyroid cells and in BRAFV600E-positive papillary thyroid cancer cells.
Normal BRAF-wild-type thyroid cells and BRAFV600E-mutated papillary thyroid cancer cells were subjected to proliferation assays and analyzed for cell death by immunofluorescence. Cell cycle status was determined using an EdU uptake assay followed by laser scanning cytometry. In addition, expression of proteins within the MAPK signal transduction pathway was analyzed by Western blot.
PLX4032 has potent anti-proliferative effects selectively in BRAF-mutated thyroid cancer cells. These effects appear to be mediated by the drug’s activity of inhibiting phosphorylation of signaling molecules downstream of BRAF within the pro-survival MAPK pathway. Interestingly, PLX4032 promotes the phosphorylation of these signaling molecules in BRAF-wild-type thyroid cells.
These findings support further evaluation of combinational therapy that includes BRAFV600E inhibitors in thyroid cancer patients harboring the BRAFV600E mutation.
PMCID: PMC3976539  PMID: 24673746
Thyroid cancer; BRAFV600E mutation; PLX4032; MAPK signal transduction pathway; Targeted therapy; Kinase inhibitors
21.  Rapid purification of gram quantities of β-sitosterol from a commercial phytosterol mixture 
BMC Research Notes  2014;7:182.
β-Sitosterol, a plant sterol or phytosterol, has commercial uses in the nutraceutical and pharmaceutical industries, but is also employed frequently in biological research. Phytosterols always accumulate as mixtures, and obtaining highly pure β-sitosterol in larger quantities for biological assays has been a challenge.
An improved method for the rapid purification of β-sitosterol from a commercial phytosterol extract is presented. Fractional crystallization of soybean oil yielded a soluble and an insoluble fraction. β-Sitosterol was purified by silica gel and Na-Y zeolite chromatography.
The rapid and cost effective three-step purification described here afforded β-sitosterol in gram quantities with high purity (>92%) and yield (>22%).
PMCID: PMC3974218  PMID: 24674386
β-Sitosterol; Purification; Fractional crystallization; Chromatography; Phytosterol; Zeolite
22.  Prevalence and risk factors of kidney disease in urban Karachi: baseline findings from a community cohort study 
BMC Research Notes  2014;7:179.
Chronic kidney disease (CKD) is being increasingly recognized as a leading public health problem. However, there are limited data available with respect to prevalence of CKD in Pakistan, a developing South Asian country. The study presents the baseline findings of prevalence and risk factors for adult kidney disease in a Pakistani community cohort.
A total of 667 households were enrolled between March 2010 and August 2011 including 461 adults, aged 15 and older. Mild kidney disease was defined as estimated Glomerular Filtration Rate (eGFR) ≥60 ml/min with microalbuminuria ≥ 30 mg/dl and moderate kidney disease was defined as eGFR <60 ml/min (with or without microalbuminuria).
The overall prevalence of kidney disease was 16.6% with 8.6% participants having mild kidney disease and 8% having moderate kidney disease. Age was significantly associated with kidney disease (p < 0.0001). The frequency of diabetes, hypertension and smoking differed significantly among the three groups, i.e., no kidney disease, mild kidney disease and moderate kidney disease.
Our study results suggest that the burden of kidney disease in this population is found considerable and comparable to neighboring developing countries. We believe that these results have critical implications on health and economics of these countries and due to the epidemic of diabetes, hypertension, cardiovascular disease, smoking and association with worsening poverty, further rapid growth is expected. There is an urgent need for early recognition and prevention strategies based on risk factors and disease trends determined through longitudinal research.
PMCID: PMC3972995  PMID: 24670059
Chronic kidney disease; Pakistan; Prevalence; South Asia
23.  Identification and survival studies of Mycobacterium tuberculosis within Laboratory-Fermented bovine milk 
BMC Research Notes  2014;7:175.
Mycobacterium tuberculosis and Mycobacterium bovis are the classic agents causing tuberculosis (TB) in humans and animals respectively. Transmission of tuberculous bacteria to humans usually occurs by inhalation of aerosols containing droplets of tubercle bacilli or via consumption of contaminated foods and drinks, primarily milk. The practice of milk pooling, including from cows with TB of the udder, further exacerbates the situation by rendering the whole milk supply infective. The simultaneous presence of indigenous lactic acid bacteria (LAB) in Mycobacterium-contaminated milk is believed to confer protective effect when the milk is adequately fermented. This study assessed the effect of LAB on the viability of mycobacteria in inherently contaminated pool of raw milk during fermentation as a function of time.
Growth was obtained in the pooled raw milk culture, and identified to be M. tuberculosis. This M. tuberculosis growth was undetectable in the milk culture by day 7 as assessed by plating serial dilutions of the milk culture for up to 14 days.
Some LAB species appear to show inhibitory effect on tubercle bacilli. If proven by more rigorous, controlled experimental results regarding such effect, selected LAB (with proven safety and efficacy) may have potential applications as anti-mycobacterial agents.
PMCID: PMC3972962  PMID: 24666844
Lactic acid bacteria; Mycobacterium tuberculosis; Fermented milk; Inhibition
24.  Utilization of information and communication technology (ICT) among sexually transmitted disease clinics attendees with coexisting drinking problems 
BMC Research Notes  2014;7:178.
Alcohol misuse remains a major risk factor for contracting sexually transmitted diseases (STDs) not typically addressed in STD clinic settings. Information and communication technology (ICT) can offer new avenues to deliver evidence-based screening and treatment for problematic drinking, however, few data exists regarding the utilization of ICT among STD clinic attendees with coexisting drinking problems. The objectives of this study are to identify STD clinics attendees with hazardous drinking, to examine socio-demographic factors associated with ICT use, and to explore individuals’ interests in engaging in ICT-based health interventions.
Cross-sectional questionnaires examining alcohol consumption and ICT use were administered to 396 persons attending two non-urban STD clinics. Descriptive statistics for ICT use were calculated for both hazardous drinkers and the entire sample. Multivariable logistic regression models among hazardous drinkers identified factors significantly associated with use of each kind of ICT.
The mean age of the 396 participants was 25 years, 66% were females and 60% were African-Americans. One third of the sample met the criteria of hazardous drinking. ICT use in hazardous drinkers included 94% reporting having internet access at least monthly, 82% reporting having an email account, 85% reporting currently owning a cell phone, and 91% reporting use of any cell phone application. More than two thirds (73%) of hazardous drinkers were willing to play health-related video games during clinic waiting time, slightly higher than the entire sample (69%). Multivariable analyses indicated that younger age were significantly related to monthly internet use, and multifunction cell phone use, while being males and younger age were significantly associated with monthly video game playing.
Our study demonstrates commonality of ICT use among STD clinic attendees with hazardous drinking, indicating the viability of using ICT to assist screening and behavioural intervention for alcohol misuse in STD clinic settings. Future research is needed to demonstrate the effectiveness of ICT-based health behavioural interventions in the STD clinic settings or other venues that serve populations at high risk for substance abuse, HIV or other STDs.
PMCID: PMC3974195  PMID: 24670037
Information and communication technology (ICT); Hazardous drinking; Substance abuse; Sexually transmitted diseases (STDs); Cell phone; Email; Internet; Video games; Digital divide
25.  Candida pelliculosa endophthalmitis after cataract surgery: a case report 
BMC Research Notes  2014;7:169.
Here we report the first case of postoperative endophthalmitis due to Candida pelliculosa after cataract surgery. We describe the clinical management of this type of candida infection in the eye.
Case presentation
A 57-year-old Turk man was seen at our clinic at the end of the first postoperative month after cataract surgery. He presented with eye redness, pain and decreased visual acuity. His ophthalmologic examination revealed moderate tyndall and a mild flare in the anterior chamber. Hypopyon in the capsular bag posterior to the intraocular lens was seen in the second postoperative month. Despite topical and subconjunctival bacterial endophthalmitis treatment, there was no improvement in the clinical situation. Candida pelliculosa was isolated from a sample culture obtained from the anterior chamber. Oral fluconazole could not be administered because of increased liver enzyme levels and intravenous amphotericin B could not be administered because of an allergic reaction. Intraocular lens explantation, pars plana vitrectomy and anterior chamber lavage by rupturing the posterior wall of the microabscesses were performed. Intravitreal and intracameral amphotericin B injections were given four times in addition to surgical interventions. The patient has been followed for 2 years and his best-corrected visual acuity was 0.4 at the last visit.
Nearly 1 month after cataract surgery, a patient presented with eye redness and blurred vision, with corneal endothelial deposits, hypopyon in the capsular bag and microabscesses on the incision sites and corneal endothelium. Candida pelliculosa should be considered in patients showing these symptoms. Multiple intraocular amphotericin B (5 μg) administrations can be used safely even in cases with high sensitivity to systemic use. Rupturing the posterior wall of the abscesses on the corneal endothelium surgically with intraocular lens explantation and pars plana vitrectomy are recommended.
PMCID: PMC3974436  PMID: 24656053

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