Tehran University Medical Journal

Usage of capillary electrophoresis for common hemoglobinopathies screening

Amir Atashi

Hemoglobinopathies are most common inherited disorders in the world; approximately 7 percent of the worldwide population and 5-6 percent of population of Iran are carriers. The hemoglobin disorders inherit as autosomal recessive and are very common in the Mediterranean area and much of the Asia and Africa. The control of this inherited disorders need to genetic counseling and accurate screening by more advanced and more accurate methods. This study explains features of current Iran hemoglobin disorders, nominates the accessible methods for screening them and introduces the capillary zone electrophoresis as a rapid and more accurate method. The required data were extracted of various articles and then for good explanation, current Iran hemoglobinopathies properties were showed in the tables and electropherograms of important hemoglobin disorders in Iran population were provided for help to interpretation results of blood tests by capillary zone electrophoresis method. Hemoglobin disorders are including thalassemias and hemoglobin variants; Disruption in the production and malfunction of globin chains cause types of hemoglobin disorders. We cannot introduce one of clinical laboratory tests as critical and basic method for screening and distinguishing types of inherited hemoglobin disorders as alone. For distinguishing the types of them must be prepared enough information and data of the hemoglobin disorders and for more accurate analysis must be used simultaneously different methods as gel electrophoresis, high performance liquid chromatography, isoelectric focusing, capillary zone electrophoresis or molecular tests. The capillary electrophoresis is an accurate and rapid method for screening types of the hemoglobin disorders. Other side this method cannot analyze all of them, so must be used biochemical, biophysical and molecular methods for confirmation the results. This review showed we can use the capillary electrophoresis and HPLC as two complementary methods for hemoglobinopathies screening. We can analyze by the methods more hemoglobin disorders and decrease more laboratory errors. Moreover, we must have patient history, hematological indices, information and data of types of hemoglobinopathies. The patient history and complete blood count results as red blood cell count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration can be useful and helpful in screening the hemoglobin disorders and then distinguishing all of hemoglobin disorders.

Assessment of nanopolyamidoamine-G7 dendrimer antibacterial effect in aqueous solution

Shahram Nazari

Background: Poly(amidoamine) (PAMAM) dendrimer derivatives have been investigated for their biological applications, especially for delivery of drugs, including antimicrobial drugs to eukaryotic cells, but their effects on bacterial cells are largely unexplored. Nanotechnology and its application is one of the rapidly developing sciences. As demand of fresh drinking water is increasing, nanotechnology can contribute noticeable development and improvement to water treatment process. This study was aimed to examine synthesis and the antibacterial effect of Nanopolyamidoamine-G7 (NPAMAM-G7) dendrimer on Escherichia Coli (E. Coli), Klebsiella Oxytoca (K. Oxytoca), Pseudomonas Aeruginosa (P. Aeruginosa), Proteus Mirabilis (P. Mirabilis) and Staphylococcus Aureus (S. Aureus) from aqueous solution.
Methods: In this experimental study that has been conducted in August to December 2015 in the laboratory of microbiology of Iran University of Medical Sciences, initially dilution of 103 CFU/ml were prepared from each strain of bacteria. Then different concentrations of dendrimer (0.025, 0.25, 2.5 and 25 µg/ml) in the laboratory temperature (23-25 °C) was added to water. In order to determine the efficiency of dendrimers in removal of bacteria, samples were taken at different times (0, 10, 20, 30, 40, 50 and 60 min) and were cultured on nutrient agar medium. Samples were incubated for 24 hours at 37 °C and then number of colonies were counted.
Results: Antibacterial properties of dendrimers in aqueous solution by increasing the dendrimer concentration and contact time is directly related. At a concentration of 25 μg/ml at 60 minutes all bacteria except S. Aureus, and at 30 minutes, E. Coli and K. Oxytoca bacteria for 100% excluded. The concentration of 2.5 μg/ml at 60 minutes of bacteria, E. Coli, K. Oxytoca and P. Mirabilis are 100% excluded. All concentrations of dendrimers at different times were reduced bacteria in the PAMAM- G7 dendrimer effect on gram-negative bacteria, gram-positive bacteria was better.
Conclusion: The NPAMAM-G7 dendrimer with end amine groups exhibited a positive impact on the removal of standard strains, gram-positive and gram-negative bacteria. Therefore, it is possible to use these nanodendrimers as antibacterial in the future.

Evaluation of chemical composition, antioxidant, antibacterial, cytotoxic and apoptotic effects of Aloysia citrodora extract on colon cancer cell line

Amir Mirzaie

Background: Aloysia citrodora belongs to the Verbenaceae family of plants, a well-known herbal medicine in Iran. The aim of the present study was to investigate the chemical composition, antioxidant, antibacterial, cytotoxic and apoptotic effect of A. citrodora extract against human colon cancer (HT29) cells by using real-time polymerase chain reaction and flow-cytometry methods.

Methods: This experimental study was carried out in Islamic Azad University, East Tehran Branch, from March to September of 2014. At first, the A. citrodora chemical constituents were analyzed by gas chromatography-mass spectrometry (GC-MS) technique. In addition, antioxidant assay, antibacterial and anti-cancer effect was performed using 1,1-diphenyl-2-picrylhydrazyl (DPPH), disk diffusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods, respectively. The half maximal inhibitory concentration (IC50) value was calculated. We extracted total RNA molecules by using RNX solution, after which cDNA was synthesized. Finally, the pro-apoptotic (Bax) and anti-apoptotic (Bcl2) gene expression was performed by real-time polymerase chain reaction and apoptotic effects were analyzed using Flow-cytometry method.

Results: GC-MS analysis of Aloysia citrodora extract was shown 37 major components and the most frequent component was belonged to Spathulenol (17.57%) and Caryophyllene oxide (15.15%) The antioxidant activity of the extract was IC50= 0.6±0.03 mg/ml. The maximum and minimum antibacterial effects of extract were belonged to gram-negative and gram-positive bacteria, respectively. Cytotoxic results revealed that the A.citrodora extract have IC50= 20.1±0.78 mg/ml against colon cancer (HT29) cell line and real-time polymerase chain reaction results showed the expression level of Bax and Bcl2 was increased and decreased respectively in colon cancer cell line (3.470±0.72 (P< 0.05), 0.43±0.35 (P< 0.05)). In addition, the flow-cytometry results indicated the 38.66% apoptosis in colon cancer cell line.

Conclusion: According to the results, it seems that A. citrodora extract has potential antioxidant, antibacterial and anticancer effects and it suggested that further studies were performed for A. citrodora pharmaceutical importance.

Study of genetics, phenotypic and behavioral properties of eubacteria and archaebacteria

Mohammad Reza Pourmand

Background: The genome of the bacteria has considerable diversity in terms of sequence of nucleotide bases and change over the time. With the advancement of bioinformatics science possibility of the vast comparison to living organisms has risen. During the last two decades many information about genome sequencing of pathogenic and non-pathogenic bacteria have been published. Using this information and to find connections between them and many phenotypic characteristics and behavior of bacteria could be used in many studies. In this study we compared some of the genetic, phenotypic and behavioral properties of archaebacteria and eubacteria.

Methods: In this analytical study, genomic Information of 286 species of archaebacteria and 122 species of eubacteria were collected from the NCBI (National Center for Biotechnology Information) site which was conducted in April to June 2015. Mean of gene size, gene number, protein number and C+G content compared in the two groups of archaebacteria and eubacteria. Association of genomic characterization of bacteria with several other characteristics were analyzed using SPSS statistical software version 19 (Chicago, IL, USA). For this purpose, the Pearson correlation coefficient (Pearson), Student’s t-test and ANOVA test (One-way analysis of variance) was used. The P values less than 0.05 was considered as significant level.

Results: There was significant association between means discrepancy in two group (P= 0.01). The genome size of eubacteria and archaebacteria have significant association with some of the characteristics of bacteria, such as the C+G content, the number of proteins, genes and habitats of the bacteria (P= 0.01). As well as there was significant association between genome size and features such as number of pseudogene, mobility and type of breathing in eubacteria (P= 0.01) but not in archaebacterial (P˃ 0.05).

Conclusion: Many characteristics of eubacteria and archaebacteria are significantly associated with genomic properties. Comparison genomics of bacteria will help in identification of evolutionary origins as well as differences between different categories of bacterial.

Awareness and practice of cervical cancer and Pap smear testing in a teaching hospital in Tehran

Soheila Aminimoghaddam

Background: Cervical cancer is known to be preventable because of long period of pre-invasive stage, availability of screening tools, and effective treatments for early invasive cervical lesions. Screening is main measures to prevent the disease and Pap smear is a screening strategy for cervical cancer. Current paper aimed to evaluate levels of awareness and practice regarding Pap smear screening among women aged between 20 to 65 years in Tehran (Iran).

Methods: This was a descriptive-analytical study conducted in Tehran City of Iran in 2015 at Firoozgar Hospital. The research population included all married, widowed and divorced women aged 20-65 years. Data analysis was performed using the Pearson correlation and Student’s t-tests in SPSS, ver. 23 (Chicago, IL, USA).

Results: Among 90 individuals who have fill questionnaire completely, 66.6% subjects had Pap smear tests. 40% of the individuals aged between 30 to 39 and the education level is distributed equally between Intermediate, Diploma and graduate and only 3 percent of them, continue their education to higher level. There was a significant relationship between the awareness of Pap smear and educational level (of both wives and husbands). The people who have graduate degree, have the best awareness. Working women revealed higher level of awareness about Pap smear. Shame and fear of taking the cancer were the most common reasons which lead to avoidance in doing the test by the women, while the most encouraging factors for performing the test were the information mostly provided by physicians and after that, the information provided by friends.

Conclusion: The awareness of Pap smear test which was measured by weighting different questions in the questionnaire by experts, prove that the women aged above 39, have an average level of awareness of Pap smear test. Due to high prevalence of cervical cancer and prolonged pre invasive course, role of Pap smear for early diagnosis necessitate the use of proper and inexpensive instructional methods to increase awareness in women about cervical cancer and preventive strategies.

Comparison of duloxetine and methylphenidate in the treatment of children with attention-deficit/hyperactivity disorder

Nastaran Habibi

Background: Attention-deficit/hyperactivity disorder (ADHD) is a common and mostly chronic mental health condition that affects children, adolescents, and adults. Stimulants and atomoxetine are first-line agents for the treatment of ADHD. Despite the impressive track record of stimulants in the treatment of ADHD, they fail in 25% of patients due to lack of efficacy or the emergence of unwanted side effects. Accordingly, this study carried out to compare efficacy and safety of duloxetine (a serotonin and norepinephrine reuptake inhibitor) and methylphenidate (a short acting stimulant) in the treatment of children with attention-deficit/hyperactivity disorder.

Methods: Twenty-four children diagnosed with ADHD participated in this 6 weeks open clinical trial. Patients were between 6 to 11 years old that had been referred to psychiatry clinic at Akhavan and Rofide Medical and Rehabilitation Center in Tehran from September 2012 to July 2014. Diagnosis was made by two child psychiatrist according to DSM-IV TR criteria. Thirteen patients received duloxetine and others received methylphenidate. Conner’ parent rating scale-revised-short form (CPRS-RS) and ADHD-rating scale (ADHD-RS) were used at the beginning and then each two weeks to assess efficacy of treatment. Routine laboratory tests and electrocardiogram (ECG) was carried out in the beginning and end of the trial.

Results: Twenty children with ADHD completed the study (Ten in methylphenidate and ten in duloxetine group). In both groups, scales of CPRS-RS and ADHD-RS were reduced from baseline to endpoint, but this reduction in methylphenidate group was significantly greater than duloxetine group (P= 0.000). The most common side effect was gastrointestinal problems in duloxetine group and anorexia in methylphenidate group. No serious side effects and no changes in laboratory and ECG indexes were seen in both groups.

Conclusion: Duloxetine is not efficacious as well as methylphenidate in treatment of children with ADHD. Although more research are needed to achieve more accurate results.

Association between serum homocysteine and subclinical atherosclerosis markers in subjects with and without diabetes mellitus

Hossein Fakhrzadeh

Background: The relationship between serum homocysteine levels and cardiovascular diseases has been elucidated since many years ago. In this study, the association between serum levels of homocysteine, folic acid, and vitamin B₁₂with the pulse wave velocity and Buckberg index or subendocardial viability ratio was assessed in individuals with diabetes and also non-diabetic subjects.

Methods: In this cross-sectional study, 58 individuals with type 2 diabetes and 36 non-diabetic people, from April to October 2013 were enrolled in Dr. Shariati Hospital affiliated to Tehran University of Medical Sciences. Anthropometric and blood pressure measurements were performed with standard methods. Fasting serum glucose, total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, Triglyceide, A₁C, vitamin B₁₂, folic acid and serum homocysteine levels as well as, highly sensitive complement-reactive protein (hs-CRP) were measured. Artherial stiffness was assessed by calculating pulse wave velocity and aortic agumentation index via Sphygmocor. In addition, Buckberg index (Subendocardial viability ratio) was assessed by dividing myocardial oxygen supply to dimand expressed as percent. The normality of distributions was evaluated by Kolmogorov-Smirnov test and linear regression models were utilized to detect associations.

Results: Diabetic and non-diabetic subjects differed in terms of age, history of hypertension, serum levels of homocysteine, and LDL-C (P< 0.05). The pulse wave velocity in subjects with diabetes and without diabetes were 60.91 m/s and 41.91 m/s, respectively (P= 0.01). After adjustment for confounding factors in multivariate regression analysis pulse wave velocity was associated with age and homocysteine levels in non-diabetic group, (β equal to 0.441 and 0.345, respectively), and it was related to age, diastolic blood pressure and serum levels of c-reactive protein in subject with diabetes (β= 0.417, 0.302, and 0.262, respectively).

Conclusion: Homocysteine levels in non-diabetic individuals were associated to sub-clinical atherosclerosis markers but we could not find this association in diabetic participants.

Assessment of educational curriculum of neonatal subspecialty in the field of challenges in neurodevelopment care

Mamak Shariat

Background: In fact, there is no doubt that medical education should be to prepare students for those clinical problems that they may encounter in their future performance. But according to the findings of previous studies in this area, one of the important priority and basic needs in education is training health workers, including physicians.

Methods: In this qualitative study focuses on the content analysis of typical (conventional content analysis) was performed. The aim of this study was to determine the needs and skills required to train neonatal subspecialists in the ability to manage vulnerable neonates problems and their families specialized in the field of comprehensive health care have driven. Based on purposive sampling, the research participants, staff and alumni of the second year and above the five-year sub-specialty in neonatology, formed by the association of neonatal diseases were chosen. Saturation as a termination criterion was applied to the collected data. Method of data collection was semi-structured interviews and focus group discussions. Reliability means the adequacy and accuracy that was measured by four methodological criteria: credibility, confirmability, transferability and dependability. All phases of study were recorded elaborately and the researchers reported all findings in the same speech research participants.

Results: Respondents consider themselves some week points in neonatal medicine education; they expressed their opinions in three categories with four subcategories as follow: "competent person knowledgeable", "weakness of the curriculum", "educational challenges", "need to review the curriculum", "the need to reform medical education system in the country", and "effective strategies for teaching".

Conclusion: Editing curriculum to teach coherent and comprehensive clinical skills in one hand, social support and health care for vulnerable children and families in other hand will improve care for vulnerable neonates.

Wernicke’s encephalopathy following hyperemesis gravidarum

Sedighe Ayati

Background: ″Wernicke’s Korsakoff″ syndrome is the most important complication of severe thiamine deficiency. The term refers to two different syndromes, each representing a different stage of the disease. Wernicke’s encephalopathy (WE) is an acute syndrome requiring emergent treatment to prevent death and neurologic morbidity. Korsakoff syndrome (KS) refers to a chronic neurologic condition that usually occurs as a consequence of WE. It is a rare complication of hyperemesis gravidarum that confusion, ocular signs, and gait ataxia are the most prevalent symptoms, respectively. Typical brain lesions of wernicke’s encephalopathy (WE) are observed at autopsy in 0.4 to 2.8 percent of the general population in the western world and the majority of affected patients are alcoholic. The prevalence of wernicke’s encephalopathy lesions seen on autopsy was 12.5% of alcohol abusers in one report. Among those who with alcohol-related death, it has been reported to be even higher, 29 to 59%. The aim of this study was to report a case of wernicke’s encephalopathy following hyperemesis gravidarum.

Case Presentation: A 28-year-old-pregnant woman in 19^th weeks of gestation referred to the hospital with hyperemesis, gait ataxia, and dysarthria before that she had hyperemesis gravidarum with weight loss and unresponsive to outpatient and inpatient medical therapy. MRI showed hyperdense lesion around thalamus which was characteristic of wernicke’s encephalopathy. Rapid improvement in patient’s condition occurred after high dose thiamine infusion.

Conclusion: In hyperemesis gravidarum, presence of either symptoms of ocular or mental disorder or ataxia must be considered to rule out and appropriate treatment of Wernicke’s syndrome which can cause maternal and fetal death.

Dysgerminoma and ovarian gonadoblastoma in Swyer syndrome

Shohre Saeed

Background: Swyer syndrome is a type of hypogonadism with 46,XY karyotype. This syndrome was named by Gerald Swyer, an endocrinologist. It leads to a female with normal internal genitalia (uterus, fallopian tubes, cervix, vagina), but instead of ovaries, they have non functional ovary (streak gonads). Also, they have absence of puberty because of gonadal digenesis. The current practice is to proceed gonadectomy once the diagnosis is made due to the fact that the risk of malignant transformation is high in dysgenetic gonad. In addition, hormonal replacement therapy after surgery is acceptable.

Case Presentation: We present a case of gonadoblastom in right ovary in a Swyer syndrome who referred to the department of Gynecology Oncology at Ghaem Hospital, Mashhad University, Iran in 2015 for evaluation of abdomino-pelvic distention. She was a 18-year-old female with 46, XY karyotype and poor secondary sexual character and normal external genitalia. She suffered of abdominal pain. In palpation of the abdomen, an irregular mobile mass was detected in left lower quadrant. The ultrasound revealed uterine size approximate dimensions 3×2 cm (infantile) and a 19 cm pelvic mass heterogeneous and multi-loculated in left side of the pelvic cavity with possible origin of the left ovary. In addition, in right pelvic fossa, a mass about 6 cm was detected. CT-Scan showed a pelvic mass with overall dimensions of 10 cm with vicinity to the left iliac vessels, modest amounts of ascities along with evidence of peritoneal dissemination (seeding). In laparotomy we observed massive ascities and a 20 cm solid mass in left ovary and a small mass in right ovary and involvement para aortic lymph node. Pathological report indicated as stage III of dysgerminoma in left ovary and gonadoblastom in right ovary.

Conclusion: This case is presented because it could have excellent prognosis if not missed opportunities of early recognizing and furthermore adequate treatment with gonadectomy.