Identifikasi Mutasi Heteroplasmi A3243g Dna Mitokondria Dan Studi Pewarisan Maternal Pada Pasien Diabetes Melitus Tipe 2

Mutasi A3243G DNA mitokondria (mtDNA) adalah mutasi subsitusi basa A ke G pada posisi 3243 gen tRNALeu. Salah satu fenotipe klinis yang disebabkan mutasi A3243G mtDNA adalah MIDD (Maternally Inherited Diabetes and Deafness) yang merupakan salah satu bentuk diabetes melitus (DM) tipe 2. Penelitian ini bertujuan untuk mengidentifikasi mutasi heteroplasmi A3243G manusia Indonesia pada pasien DM tipe 2 dan mempelajari pewarisan maternal mutasi A3243G hingga tiga generasi dengan memanfaatkan metode PCRRFLP(Polymerase Chain Reaction–Restriction Fragments Length Polymorphism), PASA (PCR-Amplification of Specific Allelle) dan PCR-SSCP (PCR-Single Strand Conformation Polymorphism). mtDNA diperoleh dari hasil lisis sampel darah. Hasil penelitian menunjukkan bahwa mutasi A3243G teridentifikasi pada 2 dari 101 pasien DM tipe 2. Mutasi heteroplasmi A3243G dapat ditunjukkan dari hasil PCR-RFLP dan PASA. Hasil RFLP menunjukkan adanya tiga fragmen, masing-masing yaitu fragmen utuh 294, 182 dan 112 pb sebagai hasil restriksi enzim ApaI. Hasil PASA menunjukkan adanya dua fragmen dengan ukuran 200 pb pada alel normal dan mutan. Mutasi A3243G yang teridentifikasi mempunyai tingkat heteroplasmi yang rendah karena tidak dapat teridentifikasi dengan direct sequencing dan PCR-SSCP (akrilamid:bis-akrilamid 49:1%). Pewarisan maternal mutasi heteroplasmi A3243G dapat teridentifikasi dengan PASA

Effect of Anti Zona Antibody on In Vitro Growth and In Vitro Maturation of Intact Follicles

Tujuan: Mengetahui pengaruh antibodi anti zona pellucida terhadap perkembangan (in vitro growth = IVG) dan pematangan (in vitro maturation = IVM) folikel. Tempat: Laboratorium biologi dan reproduksi Fakultas Kedokteran Hyogo, Nishinomiya, Jepang. Rancangan/rumusan data: Studi eksperimen pada hewan coba. Bahan dan cara kerja: Dilakukan pengambilan 80 folikel intak secara mekanik dari ovarium mencit (C57BL/6 x DBA2-F1 mice) usia 16 hari kemudian dilakukan inkubasi pada medium yang mengandung antibodi anti zona pellucida selama 8 hari. Antibodi ini diambil dari kelinci yang disuntikkan komponen ZPA dan ZPC mencit. Serum kelinci normal digunakan sebagai kontrol. Folikel dikelompokkan menjadi 4 kelompok dengan masing-masing terdiri dari 20 folikel. Kelompok 1 sebagai kontrol, kelompok 2 diinkubasi dengan serum kelinci normal, kelompok 3 diinkubasi dengan anti ZPA dan kelompok 4 diinkubasi dengan anti ZPC. Setelah 8 hari seluruh folikel dipindahkan ke medium IVM untuk dinilai perubahannya menjadi folikel antral. Hasil: Secara morfologis tidak dijumpai perbedaan bermakna pada perkembangan folikel antara kelompok kontrol dan perlakuan. Tetapi antibodi anti zona pellucida mempengaruhi perkembangan folikel antral pada kelompok perlakuan. Secara statistik dijumpai perbedaan bermakna dalam jumlah folikel antral antar kelompok 3 dan 4 dengan kelompok 1 dan 2. Pada kelompok 3 (anti ZPA) 9 dari 20 folikel (45%) berkembang menjadi folikel antral sedangkan pada kelompok 4 (anti ZPC) 11 dari 20 folikel (55%) berkembang menjadi folikel antral, dibandingkan dengan kelompok 1 dan 2, masing-masing 100% dan 85% folikel pre antral berkembang menjadi folikel antral. Kemudian seluruh folikel antral ditransfer ke medium IVM dan diinkubasi selama 16-17 jam. Pada kelompok 1, 100% folikel mengalami mucifikasi, sedangkan pada kelompok 2, 3 dan 4 masing-masing sebesar 75%, 55% dan 15% folikel mengalami mucifikasi. Setelah dilakukan denudasi germinal vesicles (GV) dijumpai sebanyak 5% pada kelompok 1,5% pada kelompok 2, dan 10% pada kelompok 4. Sedangkan pada kelompok 3 tidak dijumpai GV. Metafase 1 dijumpai sebanyak 40% pada kelompok 1,35% pada kelompok 2,50% pada kelompok 3 and 50% pada kelompok 4. Sedangkan metafase 2 dijumpai sebanyak 55% pada kelompok 1,60% pada kelompok 2, dan 40% pada kelompok 3. Tidak dijumpai metafase 2 pada kelompok 4. Beberapa oosit yang berdegenerasi dijumpai pada kelompok 2 (5%), kelompok 3 (5%) dan kelompok 4 (30%). Terdapat perbedaan bermakna dalam hal pematangan folikel (IVM) antara kelompok kontrol dan perlakuan. Kesimpulan: Antibodi anti zona pellucida mempengaruhi proses perkembangan dan pematangan folikel in vitro. Efeknya pada fertilisasi masih harus diteliti lebih lanjut. [Maj Obstet Ginekol Indones 2007; 31-4: 226-30] Kata kunci: folikel intak, perkembangan folikel (IVG), pematangan folikel (IVM), antibodi anti zona pellucida, folikel antral, mucifikasi, germinal vesicles, metafase-1, metafase-

IDENTIFIKASI MUTASI HETEROPLASMI A3243G DNA MITOKONDRIA DAN STUDI PEWARISAN MATERNAL PADA PASIEN DIABETES MELITUS TIPE 2

Mutasi A3243G DNA mitokondria (mtDNA) adalah mutasi subsitusi basa A ke G pada posisi 3243 gen tRNALeu. Salah satu fenotipe klinis yang disebabkan mutasi A3243G mtDNA adalah MIDD (Maternally Inherited Diabetes and Deafness) yang merupakan salah satu bentuk diabetes melitus (DM) tipe 2. Penelitian ini bertujuan untuk mengidentifikasi mutasi heteroplasmi A3243G manusia Indonesia pada pasien DM tipe 2 dan mempelajari pewarisan maternal mutasi A3243G hingga tiga generasi dengan memanfaatkan metode PCRRFLP(Polymerase Chain Reaction–Restriction Fragments Length Polymorphism), PASA (PCR-Amplification of Specific Allelle) dan PCR-SSCP (PCR-Single Strand Conformation Polymorphism). mtDNA diperoleh dari hasil lisis sampel darah. Hasil penelitian menunjukkan bahwa mutasi A3243G teridentifikasi pada 2 dari 101 pasien DM tipe 2. Mutasi heteroplasmi A3243G dapat ditunjukkan dari hasil PCR-RFLP dan PASA. Hasil RFLP menunjukkan adanya tiga fragmen, masing-masing yaitu fragmen utuh 294, 182 dan 112 pb sebagai hasil restriksi enzim ApaI. Hasil PASA menunjukkan adanya dua fragmen dengan ukuran 200 pb pada alel normal dan mutan. Mutasi A3243G yang teridentifikasi mempunyai tingkat heteroplasmi yang rendah karena tidak dapat teridentifikasi dengan direct sequencing dan PCR-SSCP (akrilamid:bis-akrilamid 49:1%). Pewarisan maternal mutasi heteroplasmi A3243G dapat teridentifikasi dengan PASA.Kata kunci : Mutasi A3243G, mutasi heteroplasmi, PCR-RFLP, PASA, PCR-SSCP, MIDD, direct sequencing

Case Management of Pure Gonadal Dysgenesis 46, XY (Sindrom Swyer)

Disorders of sex development (DSD) are medical conditions in which the development of chromosomal, gonadal or anatomic sex varies from normal and may be incongruent with each other. Swyer syndrome is a condition caused by pure gonadal dysgenesis 46,XY, which followed by inadequate anti-mullerian (AMH) production results in maintenance and further development of mullerian duct into uterus. Therefore, many patients who suffer from this condition initially come with chief complaint of primary amenorrhea with insufficient development of secondary sexual characteristics. Patient’s gender orientation commonly is a female, since her brain was not sufficiently exposed with androgen hormone. Management of DSD patients should be based on patient-centered approach. Therefore, overall management of DSD patients should follow according to patient’s perception regarding to her gender orientation. Herein, we reported a case of Swyer syndrome in female aged 29 years with chief complain primary amenorrhea. Chromosomal analysis was 46,XY (20 mataphase), FSH level was 31.5miu/ml, LH 10.8miu/ml, estradiol (E2) <5pg/ml, testosteron total (T) <0.0025ng/ml. Medical management for this patient has the purpose to drive her feminization process in order to improve her physical appearance. However, since there is a great tisk for having gonadal tumor development from intra-abdominal dysgenetic gonad with Y chromosome, this patient has been strongly suggested to have bilateral gonadectomy.&nbsp

The Effectiveness of Phalleria Macrocarpa Bioactive Fraction in Alleviating Endometriosis And/or Adenomyosis Related Pain

The overexpression of estrogen receptor-beta (ER-ß) and the cyclooxygenase-2 (COX-2) enzyme coupled with the absence of expression of progesterone receptors (PR) is critical to thepathogenesis of endometriosis and adenomyosis associated pain.  DLBS1442, a novel bioactiveextract of Phaleria macrocarpa, exerts its action by downregulating the overexpressed ER-ß andCOX-2 products and up-regulating PR gene expression. This pilot study was conducted to evaluatethe effectiveness of DLBS1442 treatment in alleviating endometriosis- and/or adenomyosis-relatedpain. Ten endometriosis and/or adenomyosis patients were recruited consecutively at YasminClinic Dr. Cipto Mangunkusumo General Hospital in January - March 2013. Pain associated withmenses, including pre-menstrual pain, dysmenorrhea, dyschezia and dysuria, was measuredusing the visual analog scale (VAS) at each of the next three menstrual cycles. Patients reportingone or more pain symptoms with a VAS score = 4 were given 100 mg of DLBS1442 three timesdaily for 12 weeks. VAS score reduction was noted in the first post-treatment menstrual cycle(approximately 5.3 weeks after treatment initiation) and VAS scores continued to decline overthe final two cycles. DLBS1442 was effective in alleviating endometriosis- and/ or adenomyosisrelatedpain, as demonstrated by early pain reduction as evaluated using the VAS

Response of Bitot\u27s spots to a single oral 100,000- or 200,000-IU dose of vitamin A

PURPOSE: A randomized, controlled clinical trial was conducted in Indonesia to study the response of Bitot\u27s spots to a 100,000-IU dose of vitamin A, which is known to be associated with fewer acute side effects than the currently recommended 200,000-IU dose. METHODS: A total of 114 children (ages 13 to 59 months) with Bitot\u27s spots were given an ocular examination; serum retinol concentration was measured, and the relative dose response test carried out. After administering one 100,000- or 200,000-IU oral dose of vitamin A, ocular examinations were repeated weekly for seven weeks and then biweekly for 20 more weeks, or until lesions were healed on two consecutive examinations. RESULTS: Either dose of vitamin A was similarly effective in healing Bitot\u27s spots. The most important factor in predicting responsiveness to treatment was baseline serum retinol concentration: children with lower pretreatment concentrations were more likely to have responsive lesions. No child had a relapse within the first three months after treatment. However, by six months, children who had received the higher dose were 82% less likely to have a relapse compared with children who had received the lower dose. CONCLUSIONS: Although either a 100,000- or 200,000-IU dose of vitamin A is similarly effective in healing Bitot\u27s spots, a 200,000-IU dose provides longer protection. This benefit justifies the higher rates of transient mild side effects associated with the 200,000-IU dose. The current 200,000-IU dose of vitamin A recommended by the World Health Organization for prophylactic dosing should not be reduced

A 210-μmol dose of vitamin A provides more prolonged impact on vitamin A status than 105 μmol among preschool children

A randomized controlled clinical trial was conducted to determine the relative protection afforded by two large doses of vitamin A against subclinical vitamin A deficiency among 345 preschool children. At baseline, children either had or were at high risk of developing non-corneal xerophthalmia. Vitamin A status was assessed by the relative dose response (RDR) test, serum retinol concentration, and ocular examination before and 3 and 6 mo following one oral dose of 105 μmol or 210 μmol of vitamin A. At 3 and 6 mo, mean serum retinol concentration was significantly higher in the 210-μmol group than in the 105-μmol group. The proportion of children with a positive RDR did not differ between groups at 3 mo, but by 6 mo there were three times more children positive in the 105-μmol group. Most of the observed difference was confined to children with xerophthalmia at baseline. The relative benefit of the 210-μmol dose was related to baseline vitamin A status. The current World Health Organization recommended prophylactic dose of 210 μmol seems appropriate

The Effectiveness of Phalleria macrocarpa Bioactive Fraction in Alleviating Endometriosis and/or Adenomyosis Related Pain

The overexpression of estrogen receptor-beta (ER-ß) and the cyclooxygenase-2 (COX-2) enzyme coupled with the absence of expression of progesterone receptors (PR) is critical to thepathogenesis of endometriosis and adenomyosis associated pain.  DLBS1442, a novel bioactiveextract of Phaleria macrocarpa, exerts its action by downregulating the overexpressed ER-ß andCOX-2 products and up-regulating PR gene expression. This pilot study was conducted to evaluatethe effectiveness of DLBS1442 treatment in alleviating endometriosis- and/or adenomyosis-relatedpain. Ten endometriosis and/or adenomyosis patients were recruited consecutively at YasminClinic Dr. Cipto Mangunkusumo General Hospital in January - March 2013. Pain associated withmenses, including pre-menstrual pain, dysmenorrhea, dyschezia and dysuria, was measuredusing the visual analog scale (VAS) at each of the next three menstrual cycles. Patients reportingone or more pain symptoms with a VAS score = 4 were given 100 mg of DLBS1442 three timesdaily for 12 weeks. VAS score reduction was noted in the first post-treatment menstrual cycle(approximately 5.3 weeks after treatment initiation) and VAS scores continued to decline overthe final two cycles. DLBS1442 was effective in alleviating endometriosis- and/ or adenomyosisrelatedpain, as demonstrated by early pain reduction as evaluated using the VAS. Keywords: DLBS1442, dysmenorrhea, endometriosis, adenomyosis Efektivitas Ekstrak Bioaktif Phaleria macrocarpa pada Masalah NyeriTerkait Endometriosis dan/ atau Adenomiosis Abstrak Over-ekspresi reseptor estrogen beta (ER-ß) dan enzim siklo-oksigenase-2 (COX-2) akan menekan ekspresi reseptor progesteron (PR) di endometrium; hal tersebut penting dalampatogenesis endometriosis dan adenomiosis. DLBS 1442, ekstrak bioaktif Phaleria macrocarpa,bekerja dengan menekan over-ekspresi ER-ß dan COX-2 serta meningkatkan regulasi ekspresigen PR. Studi awal dilakukan untuk mengevaluasi efektivitas pengobatan DLBS1442 padamasalah nyeri terkait endometriosis dan/ atau adenomiosis. Sepuluh penderita endometriosis dan/atau adenomiosis di klinik Yasmin RSCM pada bulan Januari - Maret 2013 yang memiliki keluhannyeri diikutsertakan dalam penelitian ini. Dilakukan penilaian skor visual analog scale (VAS) untukkeluhan nyeri pre-menstruasi, dismenorea, diskezia dan disuria setiap 3 siklus menstruasi. Pasienyang memiliki keluhan satu atau dua gejala nyeri dengan skor VAS > 4 diberikan DLBS1442sebanyak 3 x 100 mg sehari selama 12 minggu. Penurunan skor VAS diperoleh pada siklus pertamamenstruasi pascapengobatan (sekitar 5,3 minggu setelah inisiasi pengobatan) dan penurunanskor VAS terus berlanjut setelah melewati 2 siklus terakhir pengobatan. DLBS 1442 efektif dalammengatasi masalah nyeri pada endometriosis dan/ atau   adenomiosis. Kata Kunci: DLBS 1442, dismenorea, endometriosis, adenomiosi