Cytogenetic Analysis for Research and Services

AbstractThat the correct chromosome number in man is 46 was first recognized by Tjio and Levan in 1956. Perhaps few Indonesians know that Tjio was an Indonesian scientist studying in Sweden and then living in the US. Cytogenetic analyses are commonly performed to determine both structural and numerical chromosome aberration, whilst changes in chromosomes can lead to birth defects, syndromes, or even cancer.  Several chromosomal aneuploidy syndromes were identified after the establishment of various chromosome banding techniques in late 1960’s.  Specific cell culture media was found to express fragile site in the beginning of 1970’s and since then, inherited Fragile X Mental Retardation syndrome could be diagnosed.  However, some female permutation cases have been often misdiagnosed. Further molecular analysis has resolved this problem by revealing more CGG repeats in the promoter region FMR1 gene, which is related to the expression of fragile site and the severity of the diseases.In Disorder of Sex Development (DSD), early gender assignment and reconstruction surgery has been challenged because of the dilemma of gender identity development in later life. Cytogenetic analysis for the first-line gender assignment is important in newborn with DSD. Proper diagnosis with hormonal and mutation analysis should be elucidated to avoid medical, psychological, and social aspect in adult life. The most frequent genetic cases in our clinical experiences have been Androgen Insensitivity Syndrome and Congenital Adrenal Hyperplasia. Female Complete Androgen Insensitivity Syndrome (CAIS) with main symptom primary amenorrhea without cytogenetic analysis has often been diagnosed as inguinal hernia because of testicle location and size.Diagnosis and treatment of several leukemias and lymphomas, as well as some solid tumors, depend on cytogenetic analyses to demonstrate consistent, specific chromosomal aberrations. Chromosome analysis in hematologic malignancy is indicated to support diagnosis, select therapy regimen, and elaborate prognosis. Specific chromosome translocations have been identified for hematologic malignancy. The breakpoints of several of these translocations have been cloned. Several loci of oncogene have been identified and sequenced.  Molecular genetic analysis will replace cytogenetic analysis and shift the requirement for studying metaphase cells. Therefore, chromosome analysis in genetic disease and cancer should be attained with advanced molecular techniques, such as Fluorescence In Situ Hybridization (FISH) and microarray CGH analysis.  Cytogenetic analysis is still useful and applicable in genetic disease diagnosis, sexual assignment, and hematologic malignancy in the laboratory with minimal equipments. Molecular analysis as a part of health care services in Indonesia has been limited in research centers in university setting; therefore, a comprehensive diagnosis with genetic analysis has often been improbable

Genetic Analysis for the Diagnosis of Disorders of Sexual Development in Indonesia

Disorders of sex development (DSD) is defined by congenital conditions in which development of chromosomal, gonadal, or anatomical sex is atypical, while in clinical practice this term means any abnormality of the external genitalia. DSD patients have been managed by a multidisciplinary gender team in our center as collaboration between Dr. Kariadi province referral hospital and Faculty of Medicine Diponegoro University. Diagnosis should be established by specific physical examination hormonal, chromosomal and DNA studies; and imaging for most of the cases depending on indication.Since 2004 the involvement of molecular and cytogenetic analysis so far can diagnosed many of the DSD cases. Most of the genetically proven cases were Congenital Adrenal hyperplasia, Androgen Insensitivity syndrome and sex chromosomal DSD that lead abnormal gonadal development.  Many of them remain undiagnosed, further testing such as advanced DNA study should be carried out in collaboration with other center in overseas.The novel genes were found in some cases that contributed for the management of DSD.  Information for medical professionals, patients, family members and community about the availability and necessity of DSD diagnosis should be delivered to improve DSD management and patient quality of life

Genetic Analysis for the Diagnosis of Disorders of Sexual Development in Indonesia

Disorders of sex development (DSD) is defined by congenital conditions in which development of chromosomal, gonadal, or anatomical sex is atypical, while in clinical practice this term means any abnormality of the external genitalia. DSD patients have been managed by a multidisciplinary gender team in our center as collaboration between Dr. Kariadi province referral hospital and Faculty of Medicine Diponegoro University. Diagnosis should be established by specific physical examination hormonal, chromosomal and DNA studies; and imaging for most of the cases depending on indication.Since 2004 the involvement of molecular and cytogenetic analysis so far can diagnosed many of the DSD cases. Most of the genetically proven cases were Congenital Adrenal hyperplasia, Androgen Insensitivity syndrome and sex chromosomal DSD that lead abnormal gonadal development. Many of them remain undiagnosed, further testing such as advanced DNA study should be carried out in collaboration with other center in overseas.The novel genes were found in some cases that contributed for the management of DSD. Information for medical professionals, patients, family members and community about the availability and necessity of DSD diagnosis should be delivered to improve DSD management and patient quality of life

Genetic Counseling in a Couple with Primary Infertility

Background: Couples unable to conceive and bear children could feel deep disappointment, often leading to depression. Infertility is one of the main reasons couples are not able to have children. Genetic counseling role in infertility ranges from explanation about possible genetic causes of infertility, pregnancy planning, and advice for treatment. Case Presentation: A couple with 16 years of infertility was referred to the genetic clinic at National Diponegoro Hospital. The 42 years old female had previous history of diabetes mellitus, obesity, and had treatment of epilepsy/seizure 15 years ago with routine carbamazepine therapy for 2 years, while her 42 years old husband had active hepatitis B infection for 15 years. This couple underwent insemination program twice and once completed In Vitro Fertilization (IVF), both management bearing no successful implantation or viable pregnancy. Recently, she underwent a laparoscopy procedure, which gave new diagnosis of endometriosis and adenomyosis. Chromosomal examination and Methylenetetrahydrofolate Reductase (MTHFR) C677T and A1298C alleles analysis were done in our laboratory. Both individuals carried normal karyotypes and MTHFR analysis was homozygote wild type allele. Currently, this couple has accepted their conditions. They still want to bear a child although she is at a crucial ageConclusion: Infertility is a challenging and comprehensive problem. As healthcare professionals, we encounter problems not only in diagnosis and management, but also psychological and emotional dilemma. Genetic counseling is needed to solve the problems and avoid patient’s psychological distress

Focal areas of a high rate of fragile X in Indonesia: a long term follow up

Fragile X syndrome (FXS) is the most common cause of inherited intellectual disability (ID) and a leading cause of autism spectrum disorder (ASD). FXS is caused by an expansion of CGG repeats >200 in the 5′ untranslated region of the promotor region fragile X mental retardation 1 gene (FMR1), which is located on Xq27.3.  The abnormal CGG expansion leads to methylation and transcriptional silencing of the FMR1 gene, resulting in a reduction or loss of fragile X mental retardation 1 protein (FMRP) and causes long, thin, and immature dendritic spines, which lead to deficits in cognitive function, behavioral problems, and learning abilit

ANALISIS PREVALENSI DAN FAKTOR RISIKO PASIEN DENGAN ISOLATED HYPOSPADIAS DI LABORATORIUM CEBIOR

Background: Hypospadias is a common congenital anomaly characterized by the location of orificium urethra external between perineum and its normal position at the tip of the glans. It occurs in 17 per 10.000 male births. The risk factors of hypospadias are still unclear, so hypospadias is still difficult to prevent. Aims: To analyze prevalence and risk factors isolated hypospadias in patients referred to the Center for Biomedical Research (CEBIOR). Methods: Two hundred and forty nine patients were registered during the period January 2005-April 2015. Prevalence and risk factors were analyzed descriptively using secondary data and risk factors were also analyzed with Chi-Square test. Results: The declining number of patients with isolated hypospadias in CEBIOR was found because of the movement process of laboratory and alteration system of assurance. All of patients with isolated hypospadias had male karyotype 46, XY (100%). Based on the anatomic position of orificium urethra external, most of isolated hypospadias cases were penile hypospadias (60.69%). An increased risk of severe isolated hypospadias was found in mother aged older than 35 years old (PR: 1.976, 95% CI: 1.048-3.726). Parity, low birth weight, smoking father, pesticide exposure, mosquito repellant incense exposure, hormonal contraceptive use, and taking certain medication were not associated with increased severity of isolated hypospadias (p>0.05). Conclusions: The number of patients with isolated hypospadias in CEBIOR was decreased recently. The advanced maternal age was the risk factor of severe isolated hypospadias. Keywords: Isolated hypospadias, prevalence, parity, maternal age, birth weight, environmental facto

Factors Affecting Parents' Acceptance towards Children with Familial Intellectual Disability (ID)

Background: Familial intellectual disability (ID) is a condition where two or more family members are affected ID, which may influence the whole family well-being. Children with intellectual disability often receive negative response from the society, which may trigger different reactions from the parents, such as denial or neglect of their child. Besides, most parents give more attention and provide the best care for their children. Factors that may influence parents’ acceptance towards children with familial ID are social support, religious coping, supporting facilities, family income, education, mothers’s age, and other significant factors.Objective: This study was aimed to analyze factors that affect parents’ acceptance towards children with familial intellectual disabilities (ID).Methods: This was an analytic observational study with cross sectional approach. Data were collected using interview with 20 mothers of familial intellectually disabled children including demographic data, pedigree construction, using Parental Rejection Questionnaire (PARQ), Brief Arab Religious Coping Scale (BARCS), Social Support Questionnaire Short Form (SSQSR) and Supporting Facilities Questionnaires. Data was analyzed using multivariate logistic regression.Results: Parents’ acceptance was significantly affect by social support (p0.05).Conclusion: Social support has influenced parent’s acceptance of their familial ID Childre

GAMBARAN GEN JAK2 PADA PENDERITA POLISITEMIA VERA DI LABORATORIUM CEBIOR

Background: Polycythemia vera ( PV ) is one of the myeloproliferative malignancies. The Jak2V617F mutation is found in approximately 96 percent of people with PV. JAK2 gene mutation results in the production of an activated JAK2 protein, which appears to increase the production of blood cells. The identification of Jak2V617F mutation on Polycythemia Vera assists the doctors in diagnosing and determining the target of therapy. This molecular diagnosis is quite common in the developed countries but in Indonesia only Center for Biomedical Research (Cebior) Medical Faculty of Diponegoro University conducts it. The study aims to know the distribution of Jak2 gene on polycythemia vera’s patients at Cebior in Semarang from May 2012 to April 2015. Subject and method: The research employs descriptive retrospective observational cross-sectional design. The subject of the study were all patients suspected PV who were reffered to Cebior for Jak2V617F examination during May 2012 to April 2015. Result: Out of 138 patients who were referred for Jak2V617F examination at Cebior, 50 patients with referral diagnosis of PV and the incidence of PV increased every year . The result of the examination showed that 30 patients (60%) were positive of Jak2V617F. Conclusion: The identification of Jak2V617F mutation was done to 50 subjects and identified Jak2V617F mutation in 30 patients . The relatively high incidence of PV in CEBIOR further suggests the importance of the identification of Jak2V617F mutation to established the diagnoses of PV. Key words: Polycythemia vera, Jak2V617

Lower Erythrocyte GST activity in Autism Spectrum Disorder (ASD) patients compared to normal controls

Glutathione S-transferases (GST) are antioxidant enzymes that play an important role in the cellular detoxification and excretion of environmental pollutants including heavy metals. GST mu (GSTM1) and G theta (GSTT1) are known to be highly polymorphic and homozygous deletions of these genes result in the lack of enzyme activity and when combined with decreased levels of antioxidants, they have been associated with the Autism Spectrum Disorder (ASD). This preliminary study was performed to investigate the role of GSTM1 and GSTT1 polymorphisms as risk factors of ASD associated with GST activity and phenotype expression. Fifty one ASD patients and 45 controls were recruited for GSTM1 and GSTT1 genotyping while 6 ASD patients and 8 controls were assessed for GST activity. The results showed no significant differences in frequencies of GSTM1 null, GSTT1 null and combination both genotype between ASD patients and controls. However the mean erythrocyte GST activity in ASD is significantly decreased compared with controls (p = 0.043). The mean erythrocyte GST activity is lower in the severely autistic group compare to the mild to moderately autistic group, although it was not statistically significant. Further investigations are needed with a bigger sample size, analyzing multiple GST genes and GST activity determination to find out the gene susceptibility of ASD and factors that contribute to the phenotype expression of ASD

Emotional and behavioral problems in late-identified Indonesian patients with disorders of sex development

Objective: The aim of this study is to investigate emotional and behavioral problems among Indonesian patients with disorders of sex development (DSD) who recently came under clinical management. As diagnostic proce-dures and treatment had been delayed, patients progressively developed ambiguous bodies, difficult to conceal from outsiders. Method: We compared 118 Indonesian patients with DSD aged 6–41 years (60 children, 24 adolescents, 34 adults) and 118 healthy control subjects matched for age, gender, and residential settings. We used the Child Be-havioral Checklist (CBCL), Youth Self-Report (YSR), and Adult Self-Report (ASR) to examine differences between patient and control groups as well as differences within patients groups. Results: On the CBCL, parents of young children with DSD reported significantly more emotional and behavioral problems than parents of matched control. Parents of daughters with CAH reported that their daughters withdrew themselves from social interactions. On the ASR, adults with DSD reported significantly more internalizing problems than controls, particularly anxiety and depression. No other differences in emotional functioning were found across different diagnostic groups. Conclusions: Indonesian patients with DSD who were untreated for most of their lives suffered more emotional and behavioral problems than matched controls. Differences and similarities between our findings and observations in patients from Western countries will be discussed