Background. During the early embryological development of the face, complex orofacial failure results in a non-syndromic cleft lip and palate (NS CLP). The interferon regulatory factor 6 gene (IRF6) rs2235371 is a non-synonymous polymorphism that is one of the strong candidate genes associated with NS CLP.Objectives. The purpose of this study was to determine IRF6 rs2235371 as a risk factor for NS CLP and its phenotypes, including complete unilateral cleft lip and palate (CUCLP), bilateral cleft lip and palate (BCLP), cleft lip only (CL), and cleft palate only (CP), as well as to examine the effect of the polymorphism on the IRF6 mRNA expression levels among the Deutero-Malay race in Indonesia. Material and methods.This study used a case-control design and enrolled 264 samples, including 158 NS CLP cases (42 NS CUCLP, 34 NS BCLP, 33 NS CL, and 49 NS CP) and 106 control subjects. DNA was extracted from venous blood, and then subjected to polymerase chain reaction (PCR) and sequencing. The odds ratio (OR) was used to determine the risk factor for NS CLP and its phenotypes. The Livak, Kruskal-Wallis and Mann-Whitney U tests were used to determine mRNA expression levels in the oral epithelium, followed by real-time quantitative PCR (RT-qPCR).Results. Among all of the NS CLP cases, in the NS CP phenotype, OR for the A mutant allele and the GA genotype was 2,492 (p = 0.017) and 2,114 (p = 0.048), respectively. The IRF6 mRNA expression level of the GA genotype was higher in the NS CP subjects as compared to the GG genotype (p = 0.031). Conclusions.The IRF6 rs2235371 polymorphism is associated with the NS CP phenotype in Deutero-Malay patients from Indonesia and it affects the IRF6 mRNA expression level.

Non-syndromic cleft palate only (NS CPO) is one of the most common congenital malformations that affect between 1 in 1000 - 2500 live births worldwide. The etiopathogenesis of clefts including NS CPO has been widely studied but is still poorly understood. NS CPO is considered to be a genetically complex, multifactorial disease. Based on several studies, mutations of TGFβ3 gene emerged as the strong candidate gene associated with NS CPO. The purpose of this study was to analyze the relationship between the TGFβ3 / SfaN1 gene variant and the risk of NS CPO in Indonesian patients. This study was case control design using samples from 31 NS CPO subjects and 35 control subjects. DNA was extracted from venous blood and the segment of TGFβ3 gene/ SfaN1 were amplified by using polymerase chain reaction (PCR) technique, then digestion products by SfaN1 restriction enzyme which can detect locus of gene variant / polymorphism from restriction fragment length polymorphisms (RFLP) method were evaluated. The results indicated that the gene variant as substitution of base G into A was identified in TGFβ3 gene and the frequency of heterozygous mutant GA genotype was 63,6% in NS CPO subjects and 36,4% in control subjects. The frequency of heterozygous mutant GA genotype was associated with increased risk of NS CPO (odds ratio (OR) = 2,260, 95% CI = 0,592 - 8,625). In conclusion, TGFβ3 gene / SfaN1 polymorphism can be considered as the risk factor associated with NS CPO in Indonesian patients.

Luka merupakan gangguan kontinuitas lapisan epitel kulit atau mukosa. Gingiva merupakan mukosa mulut yang seringkali mengalami luka yang dapat timbul karena trauma pada perawatan penyakit periodontal ataupun ekstraksi gigi. Membran telur ayam mengandung bahan bioaktif karbohidrat kompleks yang dapat berfungsi mempercepat perbaikan jaringan. Tujuan penelitian menganalisis pengaruh aplikasi gel serbuk membran telur ayam 10% terhadap penyembuhan luka sayat gingiva tikus galur Wistar pada hari ke 3,7, dan 14. Metode: Jenis penelitian eksperimental murni dengan jumlah sampel 30 ekor tikus galur Wistar. Luka sayat sepanjang 3 mm arah horizontal kedalaman 0,25 mm dibuat pada gingiva cekat anterior rahang atas. Kelompok uji terdiri dari 3, yaitu apikasi gel membran telur 10% (n=10), aplikasi povidone iodine 10% sebagai kontrol postitif (n=10), dan aplikasi aquades sebagai kontrol negatif (n=10). Pengamatan panjang luka seluruh sampel dilakukan pada hari ke-0, 3, 7, dan 14. Data yang diperoleh kemudian dianalisis dengan uji Kruskal Wallis dan dilanjutkan uji post hoc Mann Whitney dengan p<0,05. Hasil: Seluruh kelompok mengalami pengurangan panjang luka sayat dimulai dari hari ke 3, 7, hingga 14. Kelompok perlakuan aplikasi gel membran telur 10% dapat menyembuhkan luka sayatan lebih cepat dibandingkan kelompok kontrol positif pada hari ke-3 (1,74±0,14)mm, hari ke-7 (0,81±0,16)mm dan hari ke-14 (0,07±0,14)mm. Pengukuran panjang luka sayat di hari ke-3 dan ke-7 kelompok aplikasi gel membran telur 10% signifikan berbeda dibandingkan kontrol positif (p<0,05). Simpulan: Gel serbuk membran telur 10% dapat mempercepat penyembuhan luka sayat gingiva tikus galur Wistar pada hari ke-3, ke-7 dan ke-14.Kata kunci: luka sayat; membran telur ayam; penyembuhan luka; galur wistar Effect of 10% chicken egg membrane gel powder (Gallus gallus domesticus) on gingival wound healing time in Wistar strain rats

Objective: This study aimed to detect and analyze transforming growth factor alpha (TGFA) BamHI and RsaI gene variants which associated with the risk factor of non-syndromic cleft palate only (NS CPO) of Indonesian subject.Methods: This was case-control study using samples from 32 NS CPO subjects and 28 control subjects. DNA was extracted from venous blood, and the TGFA gene was amplified using polymerase chain reaction technique, then digestion product from TaqI and RsaI restriction enzyme was evaluated. Statistical analysis to determine significant differences of gene variant frequency among NS CPO subject and control was χ2. The odds ratio (OR) was used to determine a risk factor of NS CPO.Results: The study results showed that the TGFA BamHI gene variant was not identified in NS CPO among Indonesian but TGFA RsaI gene variant was identified. The frequency of TT/B1B1 homozygous mutant genotype was 80.0% in NS CPO subjects and 20.0% in control subjects (OR=3.857; 95% confidence interval=0.405–36.749).Conclusion: TGFA RsaI gene can be considered a risk factor of NS CPO compared TGFA BamH1 gene of Indonesian subjects.

Deoxyribonucleic acid (DNA) is a genetic material which is found in all living organisms. On the human cell or eukaryotes cell, the DNA is found in the nucleus cell and the mitochondria. The DNA arrangement on each cell in human body is the same, that is why, for the analysis meaning, DNA can be isolated from any cell in the body. The source of DNA to be analyzed usually coming from the blood sample by an injection method, such a way resulting in pain and bringing about constraint. Therefore, a study was carried out to look for an alternative of DNA isolation. The aim of this experimental study was to get an optimal DNA isolation method by using oral mucous smear method with a purpose to get a quick and easy DNA isolation. The investigation materials were in the form of samples which were taken from the oral epithelial mucous cells out of three different subjects. The epithelial cells were obtained by the oral mucous smear method which in a variation of two, four and six times of smear applications, respectively. The DNA was then isolated using buffer extraction method. The concentrations of DNA were measured by using ultraviolet spectrophotometer at 260 nm wavelength. The results of DNA isolation were analyzed by Polymerase Chain Reaction (PCR) technique. The optimal DNA isolation could be analyzed by PCR technique. The experimental results show that from three different subjects of study, DNA can be isolated optimally by oral mucous smear method with six times of smear applications.