Genetics dissection and functional studies in Hirschsprung disease

De ziekte van Hirschsprung (HSCR) is een aangeboren neurologische aandoening van de dikke darm. Patiënten met de ziekte van Hirschsprung kunnen hun ontlasting niet kwijt, ze hebben zeer ernstige obstipatie. De obstipatie wordt veroorzaakt door het ontbreken of slecht functioneren van de zenuwcellen die nodig zijn voor de peristaltiek van de darm. Het krijgen van de ziekte is erfelijk bepaald, en we kennen al 12 genen die, als ze mutaties (foutjes) bevatten, tot de ziekte kunnen leiden. Mutaties in deze genen verklaren echter maar een deel van alle patiënten. Ons onderzoek richt zich op het beter begrijpen van het zenuwstelsel van de darm en op het vinden en karakteriseren van genen en mutaties die bij kunnen dragen aan het krijgen van de ziekte. Om inzicht te krijgen in welke genen belangrijk zijn in de zenuwcellen van de darm hebben we de voorlopercellen van deze zenuwcellen uit darmen van muizen gehaald en geanalyseerd. Het leverde een lijst met genen op die essentieel zijn voor de normale ontwikkeling van het zenuwstelsel van de darm. Deze genen zijn goede kandidaat-genen voor de ziekte. Ook hebben we gezocht naar genen in gebieden die eerder waren beschreven als mogelijk belangrijk voor de ziekte. Dit leverde 1 nieuw gen op dat bijdraagt aan het ziekteproces. Tenslotte hebben we een eerder gevonden mutatie, die in 70 procent van alle patiënten voor komt, gekarakteriseerd. Ons werk heeft meer inzicht gegeven in het ziekteproces, welke genen betroken zijn en hoe mutaties bijdragen aan de ontwikkeling van de ziekte

Polymerase Chain Reaction Multiplication for the Detection of Bacterial Isolates Causing Neonatal Sepsis

BACKGROUND: Neonatal sepsis is a clinical syndrome caused by the presence of bacteria in the blood accompanied by symptoms and systemic signs of infection that occurs in the first 4 weeks of life after birth. The process of identifying pathogenic microorganisms is essential in determining the clinical condition in neonatal sepsis. AIM: The study was aimed to develop a multiplex polymerase chain reaction (PCR) method to identify bacterial isolates that cause neonatal sepsis in Indonesia with the main target of optimization of an initial design and PCR optimization. METHODS: This research is an explorative in vitro study for the optimization of an initial design and PCR methods for the detection of the main bacteria that cause sepsis neonatorum in Indonesia, namely, bacteria Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, and Pseudomonas aeruginosa. The study was conducted at the Biomolecular Laboratory of the Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia. RESULTS: Sequencing carried out and continued with Basic Local Alignment Search Tool (BLAST) sequencing results, it appears that the PCR product that has been produced conforms with the optimization targets that were previously set when doing the primer design. The level of homology found in all four species based on the results of BLAST in a sequence is as follows: K. pneumoniae 94%, P. aeruginosa 99%, E. cloacae 100%, and E. coli 100%. CONCLUSION: PCR multiplex method using design primers and conventional PCR analysis methods (using agarose gel) can be used to detect four species of bacteria that cause neonatal sepsis, namely, K. pneumoniae, P. aeruginosa, E. cloacae, and E. coli

Knowledge towards Thalassemia and Willingness to Screen among Students in Public Senior High School 3 Bandung

Background: Thalassemia carrier screening is a major preventive measure  potentially influenced by the level of knowledge, particularly in adolescents. Therefore, this study aimed to analyze the effect of health education on knowledge of thalassemia in adolescents and its association with their willingness to do thalassemia screening.Methods: A cross-sectional study was conducted using data regarding knowledge of thalassemia before and after health education sessions from 229 students at Public Senior High-School 3 Bandung. All participants attended a one-day health education in July 2019. A questionnaire was filled in to measure their knowledge regarding thalassemia before and after the session, including knowledge on etiology and definition, risk of disease, clinical manifestations, treatment, complication, prognosis, and disease prevention. Only data with complete questionnaire responses were included. These responses were scored quantitatively and analyzed for their association with participants’ willingness to screen. Results: Participants were knowledgeable concerning thalassemia before the health education session (median, range: 60.0, 25.0-90.0), and knowledge was increased significantly after the education session (median, range: 80.0, 35.0-100.0) with an increased median difference=19.99 (p-value 0.05), the willingness was slightly associated with improved knowledge regarding the risk of disease (OR: 1.02; 95%CI: 1.00-1.03; p-value <0.005). Conclusion: Health education regarding thalassemia significantly increases general knowledge of thalassemia. However, improving knowledge is not significant in influencing adolescents’ motivation to take the screening tests

erbandingan Positivitas Metode MODS, Pewarnaan ZN, dan GeneXpert untuk Mendeteksi M. tuberculosis pada Pasien Meningitis TB

Salah satu permasalahan dalam diagnosis meningitis tuberkulosis (TB) adalah rendahnya positivitas metode yang digunakan. Berbagai metode telah dikembangkan, mulai dari metode kultur MODS (microscopic observation drug susceptibility), pewarnaan Ziehl-Neelsen (ZN), hingga GeneXpert. Penelitian ini dilakukan untuk menentukan metode diagnostik terbaik mendeteksi M. tuberculosis berdasar atas nilai positivitasnya. Penelitian ini merupakan observasional analitik dengan rancangan potong lintang. Data berasal dari penelitian kohort dan ReDEFINe meningitis TB di Rumah Sakit Umum Pusat Dr. Hasan Sadikin Bandung. Data diambil dari case report form pasien dengan diagnosis meningitis TB pada periode Juli 2014–Juni 2016. Kriteria inklusi penelitian ini adalah usia ≥18 tahun dan memiliki hasil pemeriksaan MODS, pewarnaan ZN, dan GeneXpert. Analisis data menggunakan Uji Cochran’s Q dan uji lanjut dengan Uji McNemar. Sebanyak 135 subjek penelitian memenuhi kriteria inklusi dan didapatkan positivitas deteksi M. tuberculosis menggunakan MODS, pewarnaan ZN, dan GeneXpert berturut-turut adalah 46,7%; 20,0%; dan 37,8%. Nilai positivitas antara ketiga metode tersebut secara statistik berbeda bermakna dengan nilai p<0,05. Sensitivitas dan spesifisitas GeneXpert dibanding dengan MODS berturut-turut mencapai 68,3% dan 88,9%, sedangkan sensitivitas pewarnaan ZN sebesar 34,9% dan spesifisitas 93.1%. Berdasar atas nilai positivitas dan sensitivitas, hasil penelitian ini menunjukkan bahwa metode MODS masih merupakan metode diagnostik terbaik untuk meningitis TB.  Kata kunci: Diagnosis, GeneXpert, meningitis tuberkulosis, MODS, pewarnaan Ziehl- Neelsen    Comparison of Positivity Rate of MODS, Ziehl-Neelsen Staining, and GeneXpert Methods in M. tuberculosis Detection among Tuberculous Meningitis Patients One of the problems in the diagnosis of tuberculous meningitis (TBM) is the low positivity rate of the diagnostic methods. Various methods have been developed, starting from culture MODS (microscopic observation drug susceptibility) to Ziehl-Neelsen (ZN) staining and to GeneXpert. This study was conducted to determine the best diagnostic method to detect M. tuberculosis based on its positivity rate. This cross-sectional analytic observational study was conducted on TBM cohort of the ReDEFINe study in Dr. Hasan Sadikin General Hospital. Data were collected from the case reports of TBM inpatients during the period of  July 2014–June 2016. Patients ≥18 years who had complete result from MODS, ZN staining, and GeneXpert were included. Data were analyzed using Cochran’s Q test and post-hoc analysis using McNemar test. In total, 135 subjects were included in this study. The positivity rates of M. tuberculosis  detection using MODS, ZN staining, and GeneXpert were 46.7%, 20.0%, and 37.8%, respectively. The positivity rate differences among the three diagnostic methods were statistically significant with a p-value of <0.05. The sensitivity and specificity of GeneXpert compared to those of MODS were 68.3% and 88.9%,respectively. Meanwhile the sensitivity and specificity of ZN staining were 34.9% and 93.1%, respectively. Based on the positivity rate and sensitivity, the results of this study indicates that the MODS method is still the best diagnostic method for TB meningitis. Key words: Diagnosis, GeneXpert, MODS, tuberculous meningitis, ziehl-neelsen Stainin

C-reactive protein and haemoglobin level in acute kidney injury among preterm newborns

Aim To explore the possibility of C-reactive protein (CRP) and haemoglobin (Hb) in prediction and risk assessment of acute kidney injury (AKI) among preterm newborns. This is believed to be closely related to the incidences of AKI, and could be the most affordable in early detection of AKI. Methods A case control study was carried out at Dr Hasan Sadikin Hospital in Bandung with a total of 112 preterms divided into two groups: with and without AKI based on the neonatal KDIGO (Kidney Disease: Improving Global Outcomes). CRP and creatinine serum were measured within 6 hours and at 72-96 hours after birth. The routine blood count included haemoglobin, haematocrit, leucocyte, and thrombocyte in the first 24 hours of life. Results CRP increase was the most influential factor for AKI with sensitivity of 80.6% and specificity of 60.2%. An increase in CRP >0.04 had an aOR (95% CI) of 5.64 (1.89–16.84). Haemoglobin <14.5 g/dL had slightly increased aOR (95% CI) of 1.65 (1.05- 8.63) Conclusion CRP increases >0.04 and level Hb <14.5 g/dL showed acceptable as an early warning for AKI in preterm newborns

Spinocerebellar Ataxia 3 (SCA3) Patient with Peripheral Neuropathy

Spinocerebellar ataxia (SCA) 3 is a neurodegenerative disease which involves cerebellum and extra cerebellum. Neuropathy in SCA3 manifests in various ways, including axonal and demyelination lesions in sensory and motor nerves. There has not been any study that describes the peripheral neuropathy characteristics of SCA3 patients in Indonesia at the time of this publication. This paper reports a case of a 43-year-old male with known spinocerebellar ataxia 3 presented with hereditary ataxia and mild numbness in both palms since two years before. No abnormalities were found during the sensory examination. The NCS showed severe axonal demyelinating sensorimotor peripheral neuropathy. In magnetic resonance imaging (MRI), an atrophy in the cerebellum with cerebral multiple lacunar infarction was identified. Electrophysiological results revealed profound axonal lesion in peripheral nerves. To conclude, peripheral neuropathy in SCA3 represents the dominance of axonal lesions in motor nerves

Epidermal Growth Factor and Adenosine Triphosphate Induce Natrium Iodide Symporter Expression in Breast Cancer Cell Lines

AIM: This study aims to investigate the effect of ATP, EGF and combination of those two to the Natrium Iodide Symporter (NIS) expression in MCF7, SKBR3 and HaCaT cell lines. METHODS: MCF7, SKBR3 and HaCaT cell lines were treated with ATP, EGF and combination of those two for 6, 12 and 24 hours. The expression of NIS mRNA was measured through quantitative-reverse transcription-polymerase chain reaction (qRT-PCR). The NIS protein expression was confirmed by immunocytofluorescence. RESULTS: NIS mRNA was expressed in SKBR3 and HaCaT cell lines but not in MCF7. The levels of NIS mRNA expression, after treatment by epidermal growth factor (EGF), adenosine Tri-Phosphate (ATP) or the combination of both for 6 and 12 hours were not significantly different from those of untreated cells. However, the treatment by a combination of ATP and EGF for 24 hours increases the level of NIS mRNA expression by 1.6 fold higher than that of the untreated cells (1.6241 ± 0.3, p &lt; 0.05) and protein NIS expression increase significantly by the treatment than untreated cells (P &lt; 0.05). CONCLUSION: The level of NIS expression varies among the different subtypes of breast cancer cell lines. MCF7 cell line is representing the luminal A subtype of breast cancer does not express NIS. Only SKBR3 cell line express NIS and this subtype might be suitable to receive radioiodine therapy as those cells expressing NIS. A combination treatment of EGF and ATP increases the expression of NIS mRNA and protein at the membrane in SKBR3 cells