Open Access Opinion Article

From Faulty Biochemical Assumptions to Poor Diagnostic Performance: Reflections on the Failure of DNA Microarrays to Become a Reliable Clinical Diagnostic Tool

Simon Rosenfeld

Journal of Advances in Medicine and Medical Research, Page 4511-4522
DOI: 10.9734/BJMMR/2014/9661

Since their inception about two decades ago, DNA microarrays have been considered as a great hope in translational research and personalized medicine. Although DNA microarrays for gene expression profiling proved to be an indispensable tool in the laboratory settings, their applications as an instrument for clinical diagnostics have not yet produced tangible results. In this paper, we convey the idea that, apart from notoriously poor reproducibility and complexities of experimental validations, there exist other reasons hindering clinical application of DNA microarrays. These reasons are rooted in the very core of the DNA microarrays methodology, that is, in faulty biochemical assumptions underlying microarray measurements. A key premise the microarray measurements are based on is that mRNA abundances harvested from the eukaryotic cytoplasm are indicative of the activity levels of corresponding genes. There are at least two reasons why this premise is questionable. First, each transcription is supported by a number of transcription factors expressed by many genes. Due to this reason, relations between the transcription rates of genes and the mRNA abundances are the 'many-to-one', not the 'one-to-one'; therefore, abnormality in a certain mRNA abundance does not unequivocally indicate abnormality of the gene bearing its complimentary code. Second, mRNA copy numbers in cytoplasm are regulated by a number of epigenetic factors among which the post-transcriptional mRNA stability is of primary importance. Abnormal concentration of certain mRNA may result from deviant mRNA stability, thus mimicking, but having nothing to do with, presumed abnormality in transcription rates of corresponding genes. An instrument built upon so poorly understood biochemical basis can hardly serve as a reliable tool in the delicate task of diagnosis of human disease in clinical settings.

Open Access Short Communication

Age and Gender Differences in Authorship among University Hospital Physicians in Sweden, Norway and Italy (The HOUPE Study)

Lise Tevik Løvseth, Ismail Cüneyt Güzey, Ann Fridner, Daria Minucci, Olav Morten Linaker

Journal of Advances in Medicine and Medical Research, Page 4582-4590
DOI: 10.9734/BJMMR/2014/10799

Background: For decades there has been a prominent gender gap in the number publications among physicians in academic medicine. Increased recruitment of women into medicine and a new generation work force that emphasize work-life balance can contribute to narrow this gap.
Aims: The present study investigates whether younger hospital physicians may display less gender differences in authorship of scientific publications compared to those older of age.
Methodology: Baseline cross-sectional survey data among senior consultants (N=1379) working at public university hospitals in three European countries, participating in the HOUPE study (Health and Organization among University hospital Physicians in Europe).
Analysis: Chi-square tests and logistic regression analysis with probit link function.
Results: There were differences in number of publications based on country where Italy and Sweden reported a significantly higher number of first- or last authorship compared to Norway (Χ2=30.6, P<.001). Logistic regression analysis confirmed gender differences in number of publications and first-and last authorships (P<.001) across all age categories. The rate of increase in number of publications is higher for men than for women physicians.
Conclusion: These findings confirm that scientific production is still more relevant to discuss in terms of gender than generation. It is important to look at factors that are essential to career choice and faculty retention in women in particular but also among women and men in the new generation of physicians.

Open Access Original Research Article

Research of HLA II Class DRB1, DQA1, DQB1 Genetic Markers in Patients with HIV Infection and AIDS

Diana Kasjko, Vladislavs Jasinskis, Jelena Eglite, Jelena Golushko, Gunta Sture, Arthur Kalimulin, Arthur Sochnevs, Ludmila Viksna

Journal of Advances in Medicine and Medical Research, Page 4482-4500
DOI: 10.9734/BJMMR/2014/8734

Aims: To find out the most frequent associations of the HLA II class loci DRB1*, DQA1*, DQB1* with the HIV/AIDS infection.
Place and Duration of Study: The study took place in The Laboratory of Clinical Immunology and Immunogenetics (LCII) of Riga Stradiņš University (RSU), Riga, Latvia, Riga Eastern Clinical University Hospital, “Infectology Centre of Latvia”, between May 1991 and December 2004.
Methodology: We analysed the medical documentation of 2500 patients and included 1180 (888 men, 292 women, 185 of them in AIDS phase) HIV infected patients. Genomic DNA was extracted from the blood with phenol-chloroform extraction method. Low-resolution HLA typing for HLA- DRB1*; DQB1*; DQA1* was performed by polymerise chain reaction (PCR) with amplification with sequence-specific primers (SSP). PCR products were separated on 3% agarose, the amplified bands were visualized, and the HLA-DRB1; DQA1; DQB1 type was deduced.
Results: Genetic markers of immunologic alleles upon development of HIV infection – HLA-DRB1*03(17:01); 05(11:01); 07:01; HLA-DQA1*01:01; 02:01; 03:01; 06:01; HLA-DQB1* 03:02; 05:01; 03:03; 03:04, as well as resistance markers connected with slow development of HIV infection – HLA-DRB1*01:01; 04:01; 06(13:01); HLA-DQA1*01:03; 04:01; 05:01; HLA-DQB1*03:01; 03:03; 04:01-2; 06:01; 06:02-8 are located in different groups of patients. High risk markers in case of HIV infection development belonging to the following groups of alleles: HLA-DRB1*03(17:01), DRB1*05(11:01), DQA1*01:01; 03:01 un DQB1*05:01; 03:02, as well as three-loci haplotypes HLA-DRB1*03(17:01)/ DQB1*05:01/ DQA1*01:01; HLA-DRB1*05(11:01)/DQB1*03:01/ DQA1 *05:01; DRB1*01:01/DQB1*03:02/ DQA1*03:01 and DRB1*01:01/DQB1*05:01/DQA1*01:01 are determined. Resistance to HIV infection development forms in the following groups of alleles: HLA-DRB1*01:01; 06(13:01), HLA-DQB1* 03:01; 06:02-8; HLA-DQA1*01:02; 01:03, as well as in haplotypes HLA- DRB1*01:01/DQB1*06:02-8/DQA1*01:02;HLA-DRB1*06(13:01)/DQB1*06:02-8/DQA1*01:02; HLA-DRB1*01:01/DQB1*03:01/DQA1*01:02; and HLA-DRB1*06(13:01)/DQB1*06:02-8/ DQA1*01:02 in different groups of HIV/AIDS patients.
Conclusion: The prevalence of genes DRB1; DQA1; DQB1 and DRB1-DQA-DQB1 combinations in the five groups of HIV infected patients have been established. Comparative analysis was performed also in the group of healthy donors (control group). The role of the main histocompatibility complex has been established, it enables marker functions and that can be used in the additional prognostic diagnostics in case of HIV infection. The obtained results testify that upon the identification of HIV genes it is possible to understand the molecular mechanisms in case of progression of AIDS syndrome complex; this possibly can be beneficial for the determination of the clinical results of infected patients.

Open Access Original Research Article

Bacterial Profile of Diabetic Foot Ulcers of Patients Visiting a Specialist Diabetic Clinic at Komfo Anokye Teaching Hospital, Kumasi, Ghana

Ruth C. Brenyah, Richard K. D. Ephraim, Ben A. Eghan Jnr, John Asamoah

Journal of Advances in Medicine and Medical Research, Page 4501-4510
DOI: 10.9734/BJMMR/2014/6054

Aims: Diabetic foot ulcers (DFU’s) pose socio-economic challenges and are a major cause of hospital admissions and morbidity often causing suffering and poor quality of life for diabetics especially in developing world. The aim of this study was to determine the bacterial profile and antibiotic susceptibility and resistance pattern of foot ulcers of diabetics at Komfo Anokye Teaching Hospital (KATH).
Study Design: Descriptive cross-sectional. Twenty seven (27) diabetics with foot ulcers comprising 15 males and 12 females attending the diabetic clinic at KATH were recruited for this study.
Place and Duration of Study: The study was conducted at the diabetic clinic of the Komfo Anokye Teaching Hospital (KATH) between November 2006 and April 2007.
Methodology: Demographic parameters of the participants were recorded and wound swabs were obtained and cultured on blood and MacConkay agar. Organisms isolated were identified and tested for their antimicrobial sensitivity patterns using Kirby-Bauer method.
Results: The mean age of the participants, duration of diabetes and FBS were 58.2±12.0 years and 6.5±2 years and 12.3±4.0 mmol/L respectively. Two (2) patients had their toes amputated. Twenty nine (29) isolates were detected from the 27 ulcer specimens out of which 28 (97%) isolates were gram negative organisms. Proteus spp (31%) and Escherichia coli (24%) were the most common gram negative pathogens isolated in this study and Staphylococcus aureus was the only gram positive organism isolated. Ciprofloxacin (100%), ceftazidime (100%), Ceftriaxone (88.3%), gentamycin (80%) and cefotaxime (80%) were most sensitive to the isolates whereas ampicillin (0%), tetracycline (0%) and chloramphenicol (0%) were the most resistant.
Conclusions: Gram negative organisms’ highly sensitive to ciprofloxacin, ceftazidime and Ceftriaxone are the most common pathogens in DFU’s in KATH.

Open Access Original Research Article

Does the Period of Air-Drying Affect Dentin Bond Strength of Different Adhesive Systems?

Emre Nagas, Isil Cekic-Nagas, Gulfem Ergun, Pekka K. Vallittu, Lippo V. J. Lassila

Journal of Advances in Medicine and Medical Research, Page 4523-4532
DOI: 10.9734/BJMMR/2014/9132

Aims: To evaluate the effect of air-drying time of adhesives on shear bond strength of different adhesive systems.
Methodology: The occlusal surfaces of 175 mandibular third molars were ground to obtain flat dentin surfaces and then divided into three groups according to three adhesive systems used: (1) Conventional three-step adhesive (Scotchbond Multi-purpose Plus); 2) Self-etch adhesive (Adper Easy Bond) and 3) Single bottle self-etch adhesive (Scotchbond Universal adhesive). Regarding the application of adhesives before resin composite application, it was gently air-dried for 3 s in Groups 1, whereas, the adhesive was left wet in Group 2. The group 3, that was air-dried until the liquid did not move (5 s), was served as control. Following bonding of resin cement (Filtek Supreme) to dentin, the specimens were light cured for 20s with a LED. After storage in water at 37ºC for one week, the strength measurements were accomplished with universal testing machine (Lloyd LRX) until the failure occurs. Failure modes were examined using a stereomicroscope and scanning electron microscope. The data were analyzed with two-way analysis of variance (ANOVA) and TukeyHSD tests (α=0.05).
Results: The two-way ANOVA revealed that adhesive systems had a significant effect on shear bond strength values (p<0.001). However, air-drying time did not influence shear bond strength (p=0.442). Additionally, there was no interaction effect between adhesive systems and air-drying time (p=0.835).
Conclusion: The data suggests that increased air-drying time of adhesives does not significantly affect bond strength.

Open Access Original Research Article

Evaluation of Pathogenic Potentialities and Transmissibility of Aerobic and Anaerobic Bacteria in Sexually Transmitted Diseases

M. Majumdar, D. Modak, M. Ganguly, K. Mazumdar, S. G. Dastidar

Journal of Advances in Medicine and Medical Research, Page 4533-4541
DOI: 10.9734/BJMMR/2014/9927

Aims: Sexually transmitted diseases (STDs) caused by various aerobic and anaerobic bacteria have been reported from many developed and developing countries of the world. However, there is limited data available on the association of these pathogens with STDs on the Indian sub-continent. Therefore, the aim of this is to the presence of anaerobic and aerobic bacteria in sexually transmitted infections.
Study Design: Patients attending the Department of Dermatology, Venereology & Leprology, Calcutta Medical College & Hospital, suspected to be suffering from STDs, were thoroughly examined and those having typical lesions of chancroid were excluded from further work. The prevalence of different aerobic and anaerobic bacteria was determined from among the remaining patients after ruling out cases of chancres.
Place and Duration of Study: This work was carried out in the Department of Oral Medicine, R. Ahmed Dental College; Department of Dermatalogy, Venereology & Leprology, Calcutta Medical College and Hospital; Division of Microbiology, Department of Pharmaceutical Technology, Jadavpur University and Department of Microbiology, Herbicure Healthcare Bio-Herbal Research Foundation, Kolkata, for a period of seven months from April 1 to October 31, 2012.
Methodology: At least two smears were prepared from the infected ulcers of each of the patients who were not diagnosed as of chancroid. One smear was for dark ground microscopy and the other for Gram’s staining. Confirmation of chancre was by serological testing, while aerobes/anaerobes were identified following standard procedures.
Results: The organisms isolated were Staphylococcus aureus, Shigella flexneri, Shigella sonnei, Gardenerella vaginalis, Actinomyces spp, Veillonella purvula, Peptococcus heliotrinreducens, Peptostreptococcus magnus and Peptostreptococcus hydrogenalis. These were subjected to tests for their antibiotic sensitivity pattern which was followed by successful specific therapy.
Conclusion: Various Gram positive and Gram negative aerobes and anaerobes were found to be associated with STDs and these were transmissible among homosexual and heterosexual partners.

Open Access Original Research Article

Feeding Pattern and Health Challenges of Nigerian Primary School Children in a Southeastern Urban Centre

C. I. Ndukwu

Journal of Advances in Medicine and Medical Research, Page 4542-4552
DOI: 10.9734/BJMMR/2014/9942

Aims: To document the feeding pattern of public primary school children, with emphasis on the staple diet and skipped meals; and the influence of diet and certain maternal demographic characteristics on their health and nutritional status.
Study Design: This was a cross sectional descriptive study.
Place and Duration of Study: The study was done in Onitsha, Anambra state, South East Nigeria between September and December, 2010
Methodology: This was a cross sectional descriptive study with multistage sampling of 804 children aged 6 to 12years from 12 public primary schools in the area. Selection was based on the age and gender distribution in each school. Verbal and written explanation was given to the parents/care givers who were invited to the schools to respond to interviewer administered questionnaires. Information was obtained on the feeding pattern of their children and maternal demographic characteristics. The children of those who responded, thereafter, had their heights measured and were assessed for clinically obvious morbidities. Information obtained on the age of respondents was verified from the schools’ registers. Data entry and analyses were done using Microsoft Office Excel 2007 and Statistical Package for Social Sciences (SPSS) version 16.
Results: There were 406 males and 398 females with a male to female ratio of 1.02:1. Their staple diet was cassava and rice. An average of 2.5% of the children skipped one meal daily. Breakfast, which was the only meal that had a significant effect on stunted growth, (F=24.177, p value=0.002) was skipped by 2.2% of the children on the day of interview. Only 1 of the children had a fruit within 24hours of the data collection. Fourteen percent of these children were stunted and the predominant morbidity observed was dermatophytosis. Most of the mothers (82%) were engaged in petty trading and menial jobs with 23.8% of them spending less than 500naira (3USD) on daily feeding. Maternal education and occupation were significantly associated with stunting in the children; p<0.05.
Conclusion: Provision of at least 1 free school meal, preferably breakfast, is advocated for children in public primary schools. This will help to improve the daily nutritional content of the meals of these children who are obviously from poor homes.

Open Access Original Research Article

Prevalence and Risk Factors of Very Low Birth Weight in Infants Born at the Maternity and Children Hospitals in Jeddah, Saudi Arabia during 2012-2013

Amena Yahya Munshi, Omar Balbaid, Naseem Akhtar Qureshi

Journal of Advances in Medicine and Medical Research, Page 4553-4569
DOI: 10.9734/BJMMR/2014/11151

Background: The prevalence of low birth weight in infants which is associated with a large number of risk factors is increasing worldwide and is a major cause of infant morbidity and mortality.
Objective: This study aims to describe demographic, clinical and anthropometric profile of VLBW in infants, its prevalence, associated risk factors and maternal medical complications in three Maternity and Children Hospitals in Jeddah City, Saudi Arabia.

Methods: Two study designs used in this research selected two convenient samples of VLBW infants for collecting cross-sectional retrospective and prospective data. The clinical records of VLBW infants [n=387] were reviewed retrospectively for estimating the one year prevalence rate while for identifying the possible risk factors of VLBW infants, the medical files of actively admitted patients [n=61] were daily examined for a period of four months.
Results: Beside socio demographic, clinical and anthropometric characteristics of VLBW infants, this study estimated the prevalence of VLBW infants to be 3.3% along with underlying risk factors of VLBW, its comorbidities, and maternal medical complications.

Conclusion: The prevalence of VLBW infants is constantly increasing not only in Saudi Arabia but also worldwide and VLBW is associated with a variety of possible risk factors. There is a need to conduct a nationwide community-based study on the prevalence and risk factors of VLBW infants in Saudi Arabia.

Open Access Original Research Article

Delivery Safety of Women from Tribal and Non-tribal Areas, in the State of Maharashtra, India

Prakash Prakabhakarrao Doke, Shailesh Rajaram Deshpande

Journal of Advances in Medicine and Medical Research, Page 4570-4581
DOI: 10.9734/BJMMR/2014/9795

Aims: The study was conducted to obtain information about place and mode of child delivery and compare unsafe deliveries between tribal and non-tribal areas.
Study Design: This was a retrospective study with the follow up of registered pregnant women in the Primary Health Centers (PHC).
Place and Duration of Study: The study was conducted in the PHCs of the State of Maharashtra, India. They were divided into two groups, tribal and non-tribal. The study was carried out in 2009-10.
Method: A format was prepared to obtain details of delivery of children by women. The Auxiliary Nurse Midwives filled the format for all the pregnant women registered during calendar year 2008, through house to house visits in their respective areas.
Results: More than one million pregnancies were registered. There were 21.88% home deliveries, of which 6.96% were not attended by a skilled birth attendant. About 5% of the babies were delivered through Cesarean section. The proportion of home deliveries (46.79%) and the absence of a skilled birth health professional (16.19%) were significantly higher in tribal areas. Even in institutional deliveries, interventional assistance was offered to lesser extent in these areas. The relative risk of undergoing unsafe delivery was 3.25 (95%, C.I. 3.20-3.29) in tribal PHCs. The district wise analysis also supported the findings that home deliveries and overall unsafe deliveries were more in tribal districts.
Conclusion: The study concludes that substantial number of women from tribal areas is exposed to unsafe deliveries.

Open Access Original Research Article

Modified Leishman Stain: The Mystery Unfolds

K. A. Fasakin, G. R. A. Okogun, C. T. Omisakin, A. A. Adeyemi, A. J. Esan

Journal of Advances in Medicine and Medical Research, Page 4591-4606
DOI: 10.9734/BJMMR/2014/10855

Background: Current Leishman staining technique for staining thin blood films for differential leukocyte count is too time consuming to meet emergency needs in hospitalized patients with infectious and other deadly diseases. This study aimed at discovering optimal phenol: Leishman powder ratio appropriate for modified Leishman stain and finding an optimized staining reaction and facilitating rapid cellular analysis of blood without alteration in quantity and quality.
Methodology: Leishman stain was modified using phenol crystals and liquefied phenol. Various ratios of phenol and Leishman powder were experimented in absolute methanol. Fixing and staining times of staining process were manipulated to develop new staining procedures that gave optimal staining reaction on thin blood films prepared within two hours of receipt. Results were presented as photomicrographs of stained slides.
Results: 30mg and 50mg of phenol crystals or 30µL and 50µL of liquefied phenol were required to give 1:5 and 1:3 phenol: Leishman powder ratios respectively. Two modified Leishman staining techniques were developed. The first fixed thin blood films for 25 seconds and stained for 50 seconds while the second technique fixed slides for 1 minute and stained for 3 minutes. Photomicrographs of thin blood films showed excellent staining results that compared well with the conventional technique.
Conclusion: Unlike the conventional method which requires a total of 10-12 minutes, to complete the staining process, modified Leishman staining techniques require only 75.0 seconds and 4.0 minutes! Batches of blood films can be stained within a short time thus facilitating rapid diagnosis and treatment of patients.