Analisis mutasi gen pax3 dan gen mitf pada penderita sindroma waarden burg di Indonesia=Mutation analysis of PAX3 and MITF eves in Indonesian Waardenburg Syndrome family

ABSTRACT Waardenburg syndrome (WS) is an autosomal dominant disorder characterized by dystopia canthorum, pigmentary disturbance of skin, hair and eye and sensorineural deafness. The basis for the phenotypic variability observed among and between WS families is unknown. In order to understand the molecular pathology of WS, many more data of the location and the.13.e of mutation causing W S from many different families are needed. Therefore, the location and the type of mutation of WS in Indonesian families need to be identified. The previous studies of many WS families have shown that the majority of mutations are in PAX3 gene, while some are in MITF gene In this studies the PAX3 gene and the MITF gene were analyzed from one Indonesian WS family. Survey at school for hearing handicapped was done in order to discover WS individual. Following the ascertainment of family history, each person was subjected to a physical and ear examination including a hearing test. The blood from affected and unaffected individuals from this family were collected for DNA extraction using Quick Gene Isolation Kit. All exons of the PAX3 gene and MITF gene were amplified using PCR methods. To determine the presence of mutation in an exon, Single Strand Conformation Polymorphism (SSCP) methods were used, then followed by Silver Staining. If the SSCP pattern showed the difference between affected and unaffected individuals, sequencing of the suspected exon was performed. In this study, one WS family was found and considered to be W S type I (WS 1) based on their clinical feature. SSCP pattern of 9 exons of MITF gene showed that there were no differences between affected and unaffected individuals. While SSCP pattern of 8 exons of PAX3 gene showed different bands on exon 4. So, it was suggested that the mutation would occur in this exon. Sequencing of this exon revealed that the changes of one base occulted at intronic region, which is g to a transition at 60 base up stream of exon 4. It can be concluded that mutation was located at the intron, but whether these changes in the intronic region could influence the gene product, needs further elucidation. Keywords: Waardenburg Syndrome â PAX3 â congenital deafness âMITF

EXPRESSION TROPHOBLAST CELL B-CELL LYMPHOMA (BCL2) IN EARLY AND LATE-ONSET PREECLAMPSIA

Background: Anti-apoptotic Bcl-2 has an important role that is involved in the regulation of apoptosis. Abnormal apoptotic activity in preeclampsia is caused by the dysregulation of these proteins. Trophoblast antiapoptotic and proapoptotic imbalance is thought to have different influences on the process of early-onset preeclampsia and late-onset preeclampsia   Objective: to analyze the difference in Bcl-2 expression in early and late-onset preeclampsia.  Method of study: This method was conducted using a cross-sectional study in early-onset preeclampsia compared to late-onset preeclampsia in RSUP. DR. Sardjito Yogyakarta was conducted from April 2019 to June 2019 with consecutive sampling methods. Placental tissue samples were obtained from 26 pregnancies with early-onset preeclampsia and 33 pregnancies with late-onset preeclampsia. The placental expression of Bcl-2 has been investigated by immunohistochemical staining and used semi-quantitative HSCORE examination.  Results: The T-test study showed there was a significant difference in Bcl-2 expression among early onset preeclampsia (2.31±0.66 ) and late-onset preeclampsia (2.61±0.43) with p-value 0.047 (p<0.05). Bcl-2  the expression appears lower in early-onset preeclampsia with a mean value of 0.30 (CI 0.04 – 0.60).  Conclusion: Thus we can conclude the expression of Bcl-2 is considered lower in early-onset preeclampsia compared to late-onset preeclampsi

The relationship between sirtuin 1 (SIRT1) expression and tumor size, Proliferating Cell Nuclear Antigen (PCNA) expression and histological grading in rat breast carcinoma induced by dimethylbenz()anthracene (DMBA)

Controversy regarding the role of SIRT1 in pathology of cancers exists and is still under debate.SIRT1 could act as either a tumor supressor or tumor promotor. This study was conducted toevaluate the relationship between SIRT1 expression and tumor size, Proliferating Cell NuclearAntigen (PCNA) expression and histological grading in rat breast carcinoma induced bydimethylbenz(á)anthracene (DMBA). Thirty female Sprague Dawley rats were randomly allocatedinto three groups with 10 rats in each group. Group 1 as negative control was just fed thestandard food. Group 2 as vehicle control was fed the standard food and corn oil. Group 3 asinduction group was fed the standard food and induced with DMBA at dose of 20 mg/kg bodyweight (BW) in corn oil twice a week for five weeks. All rats were palpated weekly to determinethe appearance, size and location of tumors. Sixteen weeks after DMBA induction rats weresacrified and histological preparations of the breast carcinoma tissue were then processed forSIRT1 and PCNA expression examination as well as histological grading. The result showed thatSIRT1 expression was significantly higher in breast carcinoma tissue compared to normal gland(26.12 vs 0.05; p = 0.004). SIRT1-positive was observed mostly in poor histological gradecarcinomas (56.2%), and it was not observed in good histological grade carcinomas. However,there was no significantly difference between SIRT1 and histological grading (p = 0.097; r =0.285). A significant correlation between SIRT1 expression and the tumor size (p =0.009; r=0.877), as well as PCNA expression (p =0.000; r =0.790) was observed. In conclusion, thereis relationship between SIRT1 expression and tumor size as well as PCNA expression in rat breastcarcinoma induced by DMBA

Bax Expression of Throphoblast Cells did not Differ between Early and Late Onset Preeclampsia: Ekspresi Bax Sel Trofoblas tidak Berbeda antara Preeklamsia Awitan Dini dan Lanjut

Objective: To compare Bax protein expression in throphoblast cells of early and late onset PE. Methods: A cross sectional study involving 36 cases of early onset PE and 36 cases of late onset PE was conducted. Bax protein expression was evaluated from sample of placental tissue collected from the study population and calculated using H-Score. Data on age, number of parity, gestational age, body mass index was collected from the medical records. Expression of Bax was compared using Mann-Whitney test. Results: There was no difference in the clinical characteristics (age, number of parity, BMI, SBP, DBP, and MAP) between the two groups. There was no difference in the expression of Bax protein between the early and late onset PE (mean H-score early vs. late onset PE: 1.48 vs 1.46, p=0.814, Mann Whitney U test). Clinical characteristics of the study population also did not correlate with the Bax expression (R for number of parity: 0.052, age: 0.009, gestational age: -0.014, BMI: 0.063, all p values were >0.05, linear regresion). Conclusion: There is no difference in the expression of Bax protein of throphoblast cells between early and late onset PE. Keyword: apoptosis, BAX, early onset, late onset, preeclampsia   Abstrak Tujuan: Untuk membandingkan ekspresi protein Bax dalam sel trofoblas pada preeklamsia (PE) onset dini dan lambat. Metode: Sebuah studi potong lintang yang melibatkan 36 kasus PE onset dini dan 36 kasus PE onset lambat dilakukan. Ekspresi protein Bax dievaluasi dari sampel jaringan plasenta yang dikumpulkan dari populasi studi dan dihitung menggunakan skor-H. Data usia, jumlah paritas, usia kehamilan, indeks massa tubuh dikumpulkan dari rekam medis. Ekspresi Bax dibandingkan menggunakan uji Mann-Whitney. Results: Tidak terdapat perbedaan pada karakteristik klinis (usia, jumlah paritas, IMT, TDS, TDD, dan MAP) antara kedua kelompok. Tidak terdapat perbedaan dalam ekspresi protein Bax antara PE onset dini dan lambat (rata-rata H-skor PE onset dini dan lambat: 1.48 vs 1.46, p = 0.814, uji Mann Whitney U). Karakteristik klinis populasi studi juga tidak berkorelasi dengan ekspresi Bax (R untuk jumlah paritas: 0,052, usia: 0,009, usia kehamilan: -0,014, BMI: 0,063, nilai p dari semua variable tersebut adalah sebesar >0,05, dengan menggunakan regresi linier). Kesimpulan: Tidak terdapat perbedaan dalam ekspresi protein Bax pada sel trofoblas antara PE onset dini dan lambat. Kata kunci: apoptosis, BAX, onset dini, onset lambat, preeklamsi

Plasma DNA as a potential biomarker for breast cancer detection

Breast cancer is a major malignancy among Indonesian women. It is often diagnosed inthe later stages of cancer, which leads to poor prognosis and survival of the patients.This study investigated plasma DNA concentration as a potential biomarker for breastcancer. The benefit of using this detection is the cost-effectiveness and the samples canbe collected from patients using non-invasive methods. Plasma samples were obtainedfrom healthy controls (n=18) and cancer patients (n=22). Each sample was split intotwo equal portions for DNA isolation using two different methods for the NaI methodand a commercially available kit (Qiagen/ QA) method. The DNA concentration wasdetermined by using a GeneQuant spectrophotometer (Pharmacia). The t-test was usedfor statistical analysis, which was performed using the SPSS 17.0 software. Compared tothe commercial method, extraction using NaI yielded higher DNA concentration, both fromsamples of healthy controls and cancer patients (p=0,008 and p=0.000, respectively).Furthermore, regardless of the isolation method used, the plasma DNA concentrationwas higher in healthy controls than in cancer cases (p=0,032 and p=0.005, for NaIand QA methods, respectively). In conclusion, isolation methods significantly affect DNAconcentrations. The plasma DNA concentration of healthy controls is significantly higherthan those of the cancer cases, suggesting that plasma DNA concentration might be apotential biomarker for breast cancer detection with less invasive sampling method thantissue biopsies

Mammographic Density and Estrogen Receptor α Gene Polymorphism in Javanese Women

Estrogen plays important roles in breast cancer as it binds its receptor in breast tissue. The most studied variants in estrogen receptor α encoded by ESR1 gene are the ESR1 PvuII and XbaI polymorphisms, which were associated with lower sensitivity to estrogen. We determined the proportion of ESR1 XbaI and PvuII polymorphisms in Javanese woman in Yogyakarta, Indonesia and analyzed the correlation between genetic variations with mammogram density. ESR1 XbaI and PvuII polymorphisms of 50 cases and 58 controls were identified using PCR-RFLP. Breast density was assessed based on digitizer mammograms. Quantitative analysis was performed using an interactive program based on cumulus of two thresholds. Mean of density and frequencies of SNPs were compared between cases and controls to identify the association between SNPs and cancer susceptibility. Mammographic density was significantly higher in cases (52%) than controls (0.41%) (p 0.05), while the proportion between AA and GG was significantly different (p < 0.05). Haplotype 2 (CG/PX) was associated with lower sensitivity to estrogen and reflects a decrease of mammographic density. These findings were consistent with other studies that showed that ESR1 polymorphisms may affect breast cancer risk through differences in breast density.

Review of immune responses correlated with COVID-19 outcomes: the fight, debacle and aftermath in the Indonesian context.

In the current pandemic, the highly contagious nature of the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) leads to an enormous burden for the global health care system and creates challenging socioeconomic problems. Respiratory mucosa, the main entrance of SARS-CoV-2 infection, are equipped with an innate immune defense system as the initial response against infection. Activation of the adaptive immune system facilitates viral clearance as well as providing immunological memory for prevention from subsequent exposure. However, despite repeated efforts at implementing appropriate interventions, severe and fatal cases are continuing to occur and reports of recurrent cases need clarification. Host factors may contribute to the severity of the diseases while viral immune evasion is a common phenomenon leading to severe outcomes and recurrent infection. Discussions of immunological-based tests for screening, herd immunity, along with the possible advantages or potentially futile efforts of development of vaccine and alternative immunotherapy have become a part of daily household conversations. In this review, evidence of innate and adaptive immune responses or lack of them, and immunological problems relevant for SARS-CoV-2 will be summarized. Finally, perspectives for future studies especially in the Indonesian population will be sketched

BRCA1 and BRCA2 germline mutation analysis in the Indonesian population

Specific mutations in BRCA1 and BRCA2 genes have been identified in specific populations and ethnic groups. However, little is known about the contribution of BRCA1 and BRCA2 mutations to breast cancers in the Indonesian population. One hundred-twenty moderate to high risk breast cancer patients were tested using PCR-DGGE, and any aberrant band was sequenced. Multiplex ligation-dependent probe amplification (MLPA) was performed on all samples to detect large deletions in the two genes. Twenty-three different mutations were detected in 30 individuals, ten were deleterious mutations and 20 were “unclassified variants” with uncertain clinical consequences. Three of seven (c.2784_2875insT, p.Leu1415X and del exon 13–15) and two of four (p.Glu2183X and p.Gln2894X) deleterious mutations that were found in BRCA1 and BRCA2 respectively, are novel. Several novel, pathogenic BRCA1 and BRCA2 germline mutations are found in early onset Indonesian breast cancer patients, these may therefore be specific for the Indonesian population

Challenges in the Vaccination of the Elderly and Strategies for Improvement

In recent years, the elderly has become a rapidly growing proportion of the world’s population as life expectancy is extending. Immunosenescence and inflammaging contribute to the increased risk of chronic non-communicable and acute infectious diseases. Frailty is highly prevalent in the elderly and is associated with an impaired immune response, a higher propensity to infection, and a lower response to vaccines. Additionally, the presence of uncontrolled comorbid diseases in the elderly also contributes to sarcopenia and frailty. Vaccine-preventable diseases that threaten the elderly include influenza, pneumococcal infection, herpes zoster, and COVID-19, which contribute to significant disability-adjusted life years lost. Previous studies had shown that conventional vaccines only yielded suboptimal protection that wanes rapidly in a shorter time. This article reviews published papers on several vaccination strategies that were developed for the elderly to solve these problems: more immunogenic vaccine formulations using larger doses of antigen, stronger vaccine adjuvants, recombinant subunit or protein conjugated vaccines, newly developed mRNA vaccines, giving booster shots, and exploring alternative routes of administration. Included also are several publications on senolytic medications under investigation to boost the immune system and vaccine response in the elderly. With all those in regard, the currently recommended vaccines for the elderly are presented