Detection of paternally inherited fetal point
mutations for β-thalassemia in maternal plasma
using simple fetal DNA enrichment protocol
with or without whole genome amplification: an
accuracy assessment
The Journal of Maternal-Fetal & Neonatal Medicine
09 Nov 2015
علمی پژوهشی(نمایه ISI)
5
Mahboubeh Ramezanzadeh, Mansour Salehi, Ziba Farajzadegan, Sara Kamali & Rasoul Salehi
اول
Objective: To design and evaluate a noninvasive protocol for prenatal diagnosis (PND) of
b-thalassemia, using cell free fetal DNA (cff-DNA) in maternal circulation. Traditional current
PND which is mainly based on chorionic villous sampling (CVS), amplification refractory
mutation system and sequencing holds as gold standard.
Methods: Ten thalassemia trait couples with distinct mutations for the husband and wife were
included in this study. The mutations in carrier fathers were IVSI-1, IVSI-5, FR8/9 and CD44. After
maternal plasma isolation and free DNA extraction, all samples subjected to designed protocol
including DNA size separation on agarose gel, elution of DNA from the gel slices using a simple
and efficient manual purification method, with or without whole genome amplification and the
detection method was allele-specific real-time PCR.
Results: Presence or absence of the paternal mutant allele was correctly determined in all of
cases and the accuracy of designed protocol was determined 100%.
Conclusions: The protocol described here is very simple, inexpensive and easy to perform, but
with satisfactory accuracy in detection of paternal mutations in cff-DNA. Due to the risk of fetal
loss with current invasive sampling for PND, a noninvasive alternative is highly demanded in
clinical setting.
دریافت فایل پیوست
http://dx.doi.org/10.3109/14767058.2015.1095883
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Assessment of High Resolution Melt Analysis Feasibility for Evaluation of Beta -globin Gene Mutations as a Reproducible, Cost-efficient and Fast Alternative to the Present Conventional Method
Advanced Biomedical Research
2016
علمی پژوهشی
5
Mahboubeh Ramezanzadeh, Mansour Salehi, Rasoul Salehi
اول
ß-thalassemia is the most prevalent monogenic disease throughout the world. It was the first genetic disorder nominated for nation-wide prevention programs involving population screening for heterozygotes and prenatal diagnosis (PND) in Iran. Due to the high prevalence of ß-thalassemia the shift from conventional mutation detection methods to more recently developed techniques based on novel innovative technologies are essential. We aimed to develop a real time PCR based protocol using high resolution melting (HRM) analysis for diagnosis of common ß-thalassemia mutations.
Methods
Forty DNA samples extracted from peripheral blood of suspected ß-thalassemia carriers participated in this study were subjected to amplification refractory mutation system (ARMS). We then used 20 of these samples for HRM optimization. When 100% sensitivity and specificity was obtained with HRM procedure, we applied the technique for mutation detection on another remaining 20 samples as thalassemia cases with unknown mutations (detected mutations with ARMS-PCR kept confidential). Finally the HRM procedure applied on 2 chorionic villous sample (CVS) biopsied from 12 weeks gestational age pregnant women for routine PND analysis.
Result
In the first step of study Fr 8/9 (+G), IVSI-1 (G>A), IVSI-5 (G>C), IVSI-110 (G>A) and CD44 (-C) mutations were diagnosed in samples under study using ARMS-PCR technique.
Finally the HRM procedure applied on 20 unknown samples and 2 CVS The results of HRM were in complete concordance with ARMS and confirmed by sequencing.
Conclusion
The advantages of HRM analysis over conventional methods is high throughput, rapid, accurate,
cost-effective and reproducible.
دریافت فایل پیوست
DOI: 10.4103/2277-9175.180640
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Novel Multiplex Fluorescent PCR-Based Method for HLA Typing and
Preimplantational Genetic Diagnosis of b-Thalassemia
Archives of Medical Research
47 (2016)
ISI
Sharifeh Khosravi,Mansour Salehi, Mahboobeh Ramezanzadeh, Hamed Mirzaei, and Rasoul Salehia,
سوم از 5نفر
Background and Aims. Thalassemia is curable by bone marrow transplantation; however,
finding suitable donors with defined HLA combination remains a major challenge. Cord
blood stem cells with preselected HLA system through preimplantation genetic diagnosis
(PGD) proved very useful for resolving scarce HLA-matched bone marrow donors.
Methods. A thalassemia trait couple with an affected child was included in this study. We
used informative STR markers at the HLA and beta globin loci to develop a single cell
multiplex fluorescent PCR protocol. The protocol was extensively optimized on single
lymphocytes isolated from the couple’s peripheral blood. The optimized protocol was
applied on single blastomeres biopsied from day 3 cleavage stage IVF embryos of the
couple.
Results. Four IVF embryos biopsied on day 3 and a single blastomere of each were provided
for genetic diagnosis of combined b-thalassemia mutations and HLA typing. Of
these, one embryo was diagnosed as homozygous normal for the thalassemia mutation
and HLA matched with the existing affected sibling.
Conclusion. The optimized protocol worked well in PGD clinical cycle for selection of
thalassemia-unaffected embryos with the desired HLA system. 2016 IMSS.
Published by Elsevier Inc.
Background and Aims. Thalassemia is curable by bone marrow transplantation; however,
finding suitable donors with defined HLA combination remains a major challenge. Cord
blood stem cells with preselected HLA system through preimplantation genetic diagnosis
(PGD) proved very useful for resolving scarce HLA-matched bone marrow donors.
Methods. A thalassemia trait couple with an affected child was included in this study. We
used informative STR markers at the HLA and beta globin loci to develop a single cell
multiplex fluorescent PCR protocol. The protocol was extensively optimized on single
lymphocytes isolated from the couple’s peripheral blood. The optimized protocol was
applied on single blastomeres biopsied from day 3 cleavage stage IVF embryos of the
couple.
Results. Four IVF embryos biopsied on day 3 and a single blastomere of each were provided
for genetic diagnosis of combined b-thalassemia mutations and HLA typing. Of
these, one embryo was diagnosed as homozygous normal for the thalassemia mutation
and HLA matched with the existing affected sibling.
Conclusion. The optimized protocol worked well in PGD clinical cycle for selection of
thalassemia-unaffected embryos with the desired HLA system. 2016 IMSS.
Published by Elsevier Inc.
Key Words: Preimplantation genetic diagnosis, b-thalassemia, HLA typing, Multiplex fluorescent
PCR, STR markers..
دریافت فایل پیوست
http://dx.doi.org/10.1016/j.arcmed.2016.07.006
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بررسی مقدماتی تاثیر اموزش قلب سالم در سطح مدارس ابتدایی شهر بوشهر؛ پروژه قلب سالم خلیج فارس
دوفصلنامه طب جنوب
سال پنجم - شماره2
علمی پژوهشی
دکتر ایرج نبی پور، دکتر سیدرضا امامی،محمد مهدی محمدی، دکتر محمد امیری، دکتر غلامرضا حیدری، دکتر فاطمه بهرامیان، دکتر محمد هادی ظفرمند، دکتر فاطمه عزیزی ، ....،دکتر محبوبه رمضان زاده
نفر پنجم از هشتم(یازده نفرکه هفت نفر ارزش 5 دارند)
|
مارکرهای التهاب و بیماری های قلبی عروقی
کتاب خلاصه مقالات دومین کنگره پیشگیری از بیماری های غیر واگیر
سال 82
محبوبه رمضان زاده
اول از یک نفر
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پزشکی شخصی محور از بعد ملکولی تا سلامت معنوی
پنجمین همایش ملی سلامت معنوی
دکترمحبوبه رمضان زاده- دکتر اعظم امینی
اول از دو نفر
مقدمه و هدف: از زمان بقراط دید گاههای معنوی در پزشکی وجود داشته است. اما بیشتر دانشکده های پزشکی به خاطر تاکید روی اساس علمی فعالیت های پزشکی جنبه های معنوی و مذهبی را از فرهنگشان حذف کردند. با گذار از دوران مدرن و ناموفق، پزشکی به تدریج به ماهیت معنوی خود میل کرد به طوری که معنویت وارد تعریف سلامتی سازمان بهداشت جهانی شد. با این حال حداقل در کشور ما تغییری در آموزش پزشکی و یا ارائه خدمات سلامت رخ نداده است. مقاله حاضر به بررسی اهمیت ادغام سلامت معنوی در آموزش و ارائه خدمات پزشکی با نگاه پزشکی شخص محور پرداخته است.
روش: این مطالعه با روش توصیفی ونظری اجرا شده است. داده ها به روش جستجو در پایگاههای اطلاعات علمی اینترنتی و منابع علمی معتبر گردآوری شده است.
نتایج: در حال حاضر براساس مطالعات انجام گرفته این تفکر که بعد معنوی باید در مراقبت های سلامتی مورد توجه قرار گیرد در بین متخصصین علوم پزشکی آگاه به این حوزه مورد توافق است و دربعضی کشور ها در برنامه های آموزشی پزشکی وارد شده است. بیش از یک قرن پیش مفهوم هویت فردی شیمیایی (Chemical Individuality) چنین بیان شد:" عواملی که استعداد ابتلا ومصونیت به بیماری را به ما اعطاء می کنند در ساختار شیمیایی و گروههای مولکولی که ساختار کروموزم های ایجاد کننده ما را می سازند به ارث رسیده اند." امروزه می دانیم که انسان ها تفاوت های ژنتیکی اندکی دارند که مسئول تفاوت بین افراد در حساسیت یا مصونیت به بیماری، عکس العمل به عوامل محیطی و حتی در خصوصیات شخصیتی و استعداد ها ی مختلف می باشند. براین اساس پزشکی شخصی ژنومیک (Personalized Genomic Medicine) تعریف شده است که هدف آن دخالت این تفاوت ها در خدمات پزشکی به منظور ارائه خدمات دقیق و موثربه افراد می باشد.
بحث و نتیجه گیری: دردیدگاه کلی نگر برای حفظ سلامتی انسان باید به همه ابعاد او توجه کرد. در پزشکی شخص محور اعتقاد براین است که درمان بهینه و موثر باید براساس ویژگی های شخصی افراد باشد. با توجه به مشخص شدن نقش معنویت در سلامت وویژگی ها ی معنوی متفاوت افراد، بنابراین می توان گفت حیطه پزشکی شخص محورتابعد معنوی انسان گسترده شده است. بنابراین برای ارائه یک خدمت پزشکی مناسب لازم است تا علاوه بر آموزش فراگیران علوم پزشکی، وضعیت نیمرخ معنوی بیماربا یک شرح حال دقیق مشخص شود تا منجر به ارائه یک نسخه اختصاصی و متناسب با نیازهای فرد شود.
کلمات کلیدی: سلامت معنوی، پزشکی شخص محور، ملکولی
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Hemophilia Gene Therapy; Clinical and Molecular Aspects
J Human Gen Genom
2018 July; 2(1):e80764
علمی پژوهشی
Mohammad Zaree,1 Masomeh Sadat Sabzevari,2 Ayub Ahmadi,3 Mahboubeh Ramezanzadeh,4,* and Ali Hosseini Bereshneh5,*
چهارم از پنج نفر(نویسنده مسئول)
Abstract
Hemophilia is a coagulation disorder in which bleeding time is prolonged. There are a number of hemophilia subtypes and more
than 4,000,000 individuals are suffered worldwide. The most common types of hemophilia are type A and B in which coagulation
factor VIII and IX are defected respectively. Type A hemophilia is responsible for 80% to 85% of cases. The genes of 8 and 9 coagulation
factors located on the long arm of X chromosome and mutation in these genes causes disturbance in coagulation. This disease
is a very good target for gene therapy because if amount of protein production reaches 1% that of normal the disease phenotype
is modified. Different methods of hemophilia gene therapy include increased production of coagulation factors via insertion of
attributed genes into patient’s stem cells by vectors, or insertion of transgenes into differentiated cells with prolonged survival such
as muscle or liver cells. One of the most recent advances in hemophilia gene therapy is using induced pluripotent stem cells (iPS)
for gene transfer. Hepatocytes are very good candidates for hemophilia gene therapy due to their natural capacity for production
of coagulation factors. Myocytes are also suitable for injection of transgene because they are available and have sufficient secretory
power. Most important and useful viral vectors for hemophilia are retroviral, lentiviral, and Adeno- Associated viruses. Amongthese
only the retroviral vectors target dividing cells.
Keywords: Hemophilia, Gene Therapy, iPS
doi: 10.5812/jhgg.80764.
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Cell-free Fetal Nucleic Acid Identifier Markers in Maternal Circulation.
Adv Biomed Res
2017 Jul 28;6:89
علمی پژوهشی
PubMed
Ramezanzadeh M1, Khosravi S2, Salehi R2
اول از سه نفر
Abstract
From the discovery of cell-free fetal (cff)-DNA in 1997 so far, many studies have been performed on various aspects of cff-nucleic acid. It is undoubted that currently, invasive prenatal diagnosis progresses to the noninvasive test. However, there are many problems. One of the most challenging issues in this field is differentiation and detection of the small amount of cff-nucleic acid in maternal plasma. Many markers and methods have been used for this purpose. This review makes an attempt to review and compare the studies in the field. Six identifier markers including Y-specific sequence, polymorphisms, epigenetic difference, DNA size difference, fetal mRNA, and microRNA as well as the advantages and disadvantages of each marker are discussed. This review provides a relatively perfect set on cff-nucleic acid biomarkers in various physiological and pathological status of pregnancy, helping to review and compare the prior obtained results, and improving designation in future studies.
KEYWORDS:
DNA; RNA; marker; pregnancy; prenatal diagnosis
doi: 10.4103/2277-9175.211800. eCollection 2017.
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بررسی اثر بخشی معنویت درمانی بر الکسی تایمی بر سرطان پستان با گیرنده های استروژن و پروژسترون
مطالعات ناتوانی
89- 1398
علمی پژوهشی
isc
حسینی- انصاری- روانبد- رضایی- رمضان زاده
5 از 5
دریافت فایل پیوست
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اثر بخشی معنویت درمانی بر سبک زندگی ارتقا دهنده سلامت در افراد مبتلا به سرطان پستان با گیرنده استروژن و پروژسترون
فصلنامه پرستار و پزشک در رزم
24- 1398
علمی پژوهشی
حسینی- انصاری- روانبد- رضایی- رمضان زاده
5 از 5
دریافت فایل پیوست
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ارتباط بین سلامت عنوی و پیشرفت تحصیلی دانشجویان علوم پزشکی بوشهر
آموزش در علوم پزشکی
20- 1399
علمی پژوهشی
ISC
برنا- موسوی نسب- جوهری- رمضان زاده
4 از 4 - نویسنده مسوول
دریافت فایل پیوست
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ارتباط بین سلامت معنوی و پیشرفت تحصیلی دانشجویان دانشگاه علوم پزشکی بوشهر
آموزش در علوم پزشکی
20- 1399
علمی پژوهشی
ISC
برنا- موسوی نسب- جوهری- رمضان زاده
4 از 4 - نویسنده مسوول
دریافت فایل پیوست
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