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Role of Human Sperm Mitochondrial Dna in Infertility

Thesis Info

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Author

Mughal, Irfan

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Animal Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9778/1/Irfan%20Mughal_QAU_2015.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727116497

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Sperm is the core of male fertility, which has to travel up to the fallopian tube for successful fertilization. Sperm motility depends on the electron transport chain producing ATPs in its mitochondria, which is a direct expression of the mitochondrial DNA (mtDNA) quality. Sperm motility is major determinant of fertility. It is already believed that mtDNA mutations are linked with infertility but the results are contradictory and previous researches are based on limited number of semen samples. Previous studies indicated a vacuum for more comprehensive study of sperm mtDNA from multiple aspects with sufficient number of carefully selected subjects to find more concrete findings. This case control study was designed on these hard facts to find association of sperm mtDNA deletions with fertility. We hypothesized that sperm mtDNA deletions have significantly associations with human male infertility. We collected 355 human semen samples (following WHO protocols), 74 samples normal controls (produced at least one child) and 281 infertile patients. Infertile samples were further classified into five groups, asthenozoospermia (As), oligozoospermia (Oz), oligotetrozoospermia (Ot), oligonecrozoospermia (On) and oligoasthenoteratozoospermia (OAT syndrome). Infertile nfertile nfertile nfertile individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. individuals were cases of idiopathic infertility. We focused on the most coherent core part of sperm mtDNA, the COXIII subunit. DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted DNA from semen samples was extracted by modified organic protocol and the DNA quantification was carried out spectrophotometrically at 260nm/280nm. The data for control and infertile patients was compiled and analyzed with IBM SPSS version 22 (SPSS, Chicago, IL, USA) Chi Square test was applied and P value less than (P=0.05) was considered as significant. The famous deletion (9480del15bp) was analyzed with COXIIIA and COXIIIB pair of primers and data analysis revealed highly significant association between infertile and deletions (P=0.001). In second set of experiments a bigger segment (50bp upstream to 9480del15) was explored with COXIIIC and COXIIIB primers of which the frequency of mutations was significantly higher in OAT samples (P=0.038). In third experiment multiple deletions were amplified simultaneously in larger segment by long PCR with MT1A and MT3 primers data analysis revealed highly significant association (P=0.001). Multiple mtDNA mutation had direct relation with poor semen quality and maximum incidence of mutations was identified in OAT syndrome. Sequencing analysis of COXIII indicated a polymorphism at position 9505, transition of C>G in all sample analyzed. Another transverse of codon at position 9526 marked by the change of C>A was discovered. This study demonstrated that sperm mtDNA deletions have significant roles in the diminution of fertility by deteriorating various sperm motility parameters. All four set of experiment revealed similar pattern of deletions associated to male infertility, highest incidence found in OAT group. The results are conclusive of “the male infertility is significantly associated to sperm mtDNA deletions.” We estimated that simple semen analysis parameters might be predictive of sperm mtDNA quality.
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خود شناسی خدا شناسی ہے

خودشناسی خدا شناسی ہے
اللہ تعالیٰ نے ہر چیز جو پیدا فرمائی ہے اس کی ایک شناخت ہے، سورج کی ا یک شناخت ہے کہ وہ مشرق سے طلوع ہوتا ہے اور مغرب میں غروب ہوتا ہے۔ ستارے رات کو چمک کر شب دیجور کے لیے ضیاکا سامان بہم پہنچاتے ہیں اور مسافرانِ شب کے لیے خضر راہ بنتے ہیں تو یہ ستاروں کی ایک شناخت ہے، فلک بوس پہاڑ اور جبال شامخہ کا ایک طویل سلسلہ بھی اپنی شناخت رکھتا ہے۔ الغرض دنیا و مافیہا ہر چیز اپنی شناخت رکھتی ہے۔ جس کے باعث اس کا وجود قائم ہے۔
انسان کی بھی ایک شناخت ہے کہ وہ حیوان ناطق ہے اور ذوی العقول ہے، اس کی چال ڈھال، اس کی نشست و برخاست، اس کا قیام وقعود، اس کی گفت وشنید اس کو دیگر مخلوقات سے ممتاز کرتی ہے، اور پھر قرآن پاک اس کو اشرف المخلوقات کا لقب عطا فرما کر انسانیت کی معراج پرمتمکن بنادیتا ہے، اس سے بڑھ کر مسلمان جو اسلام کے زیور سے مرصعّ ہے، جس نے دینِ اسلام کا تاج اپنے سر پر سجایا ہوا ہے، جس نے عشق مصطفی صلی اللہ علیہ و آلہٖ وسلم کی خلعت ِفاخرہ زیب تن کی ہوئی ہے اس کی بھی ایک شناخت ہے، اور وہ یہ کہ ارشادِ رسالت مآب صلی اللہ علیہ و آلہٖ وسلم ہے ’’مسلمان وہ ہے جس کے ہاتھ اور زبان سے دوسرے مسلمان محفوظ رہیں‘‘ یعنی اسلام کے دعویدار تو بے شمار ہیں لیکن دینِ اسلام کے احکام پر عمل پیرا حقیقت میں وہی ہے جو مذکورہ حدیث پاک کے مطابق اپنی شناخت رکھتا ہے۔
کرسی عدالت پر بیٹھ کر فیصلہ سنانے والے منصف کی ایک شناخت ہے۔ شفاخانوں میں موجود مریضوں کے علاج کرنے والا مسیحا کی ایک شناخت ہے، سرحدوں پر مامور محافظ مجاہد کی ایک...

اﻷداء اﻟﻤﺄﻣﻮر ﺑﻪ و ﺗﻄﺒﻴﻘﺎﺗﻪ اﻟﻔﻘﻬﻴﺔ

In the Islamic Law i. E. Quran and Sunnah the above mentioned topic has been discussed clearly and repeatedly to avoid any ambiguity in dealings in our daily life and agreements made between two parties or governments national or international levels. After intensive study I discussed and elaborated the said issue referring to the Qura’nic verses and sayings of the Prophet (SAW). The matter is of great importance; hence the Muslims Jurists have also given their valuable opinions in accordance with the Islamic Law which have been incorporated also to solve the issue. If we act upon these verdicts, we will be able to select the best among ourselves and form an ideal government and will discharge our duties honestly, and eventually our every act will show our responsibility to perform our duties and to give due share to the right person.

Study of Structural, Magnetic and Dielectric Properties of Ferrite/Chromite Nanoparticles

This thesis is schematically based on synthesis and characterization of cobalt chromite (CoCr2O4), maghemite (γ-Fe2O3) and nickel ferrite (NiFe2O4) nanoparticles, as well as selective coating and doping in host compounds in order to tune its structural, dielectric and magnetic properties. CoCr2O4 and NiFe2O4 nanoparticles were synthesized by sol-gel method, while γ-Fe2O3 nanoparticles were synthesised by microwave plasma technique. For chromite nanoparticles, the low temperature magnetic response of CoCr2O4 nanoparticles, magnetic and dielectric properties of Mg doped CoCr2O4 nanoparticles and magnetic properties of SiO2 coated CoCr2O4 nanoparticles have been studied in detail. X-ray diffraction revealed the cubic spinel structure of the nanoparticles. Zero field cooled and field cooled (ZFC/FC) curves revealed a paramagnetic (PM) to ferromagnetic (FiM) transition at TC = 97-100 K with conical spiral state at TS = 27 K and lock-in state at TL = 13 K. Negative magnetization is observed in the ZFC curve under 50 Oe applied field, which gets suppressed upon the application of higher field due to reorientation of the nanoparticles magnetization in the direction of applied field. The TC was shifted towards higher temperature with the application of higher field, while TS and TL remain unaffected which was attributed to strong B-B interactions which act as a frozen spins or canted spins at surface. M-H loops showed an abnormal decrease in MS which may be due to presence of stiffed/strong conical spin spiral and lock in states at low temperatures. Modified Kneller’s law showed a good fit for temperature dependent HC at higher temperature and deviated at low temperature (< 25 K) which was attributed to frozen disordered surface spins. Nanoparticles showed slow spin relaxation in both ZFC and FC protocols at 5 K, which signifies the presence of spin-glass like behavior at low temperatures. Mg doped CoCr2O4 nanoparticles showed non-monotonous trend in the average crystallite size and showed a peak behaviour with maxima at x = 0.6. The members CoCr2O4 (x = 0) and MgCr2O4 (x = 1) are FiM and antiferromagnetic (AFM), respectively. TC and TS showed decreasing trend with increasing x, followed by an additional AFM transition at TN = 15 K for x = 0.6. The system finally stabilized and changed to highly frustrated AFM structure at x = 1 due to formation of pure MgCr2O4. Dielectric parameters showed a non-monotonous behaviour with Mg concentration and were explained with the help of Maxwell-Wagner model and Koop’s theory. Dielectric properties were improved for xvii nanoparticles with x = 0.6 and is attributed to their larger average crystallite size. SiO2 coated CoCr2O4 nanoparticles showed decreasing trend of the average crystallite size and cell parameter with increasing SiO2 concentration. The decrease in average crystallite size is due to SiO2 coating which limits the growth of nanoparticles by generating more nucleation sites. All the magnetic transitions of CoCr2O4 nanoparticles shifted towards low temperatures which is due to decrease in average crystallite size. SiO2 concentration also decreased saturation magnetization (MS), which was enhanced surface disorder in smaller nanoparticles. In study of structural, magnetic and dielectric properties of ferrite nanoparticles, chromium oxide (Cr2O3) coated γ-Fe2O3 nanoparticles and NiCrxFe2-xO4 ferrite nanoparticles have been studied in detail. Simulated ZFC/FC curves exhibited large value of effective anisotropy of Cr2O3 coated γ-Fe2O3 nanoparticles as compared to bulk γ-Fe2O3 but less than bare γ-Fe2O3 nanoparticles which is may be due to weak interface anisotropy between ferrimagnetic γ-Fe2O3 core and antiferromagnetic Cr2O3 shell. Bloch’s law was fitted on T-dependent MS data and revealed the higher value of Bloch’s constant and lower value of Bloch’s exponent as compared to bulk γ-Fe2O3. Spin glass behaviour was investigated by using different physical laws for f-dependent ac susceptibility and they confirmed the presence of spin glass behaviour which is due to disordered frozen surface spins. XRD analysis of Cr doping at B site in NiFe2O4 nanoparticles confirmed the cubic spinel structure for all samples with x = 0, 0.2, 0.4, 0.8, 2.0 concentration. Saturation magnetization depicts decreasing trend with addition of Cr3+ concentration which is attributed to replacement of large magnetic moment of Fe3+ by smaller magnetic moment of Cr3+. HC reveals minimum value for NiFe2O4 nanoparticles and showed increasing trend with addition of Cr3+. This increase in HC may be attributed to change in magneto crystalline anisotropy. Dielectric constant showed increasing trend with the Cr+3 concentration due to less conductive nature of Cr as compared to Fe. In summary, a detail study of structural, dielectric and magnetic properties of chromite and ferrite nanoparticles have explored with tremendous results that will open a new insight in device applications such as automatic switching, magnetic memory and targeted nanotherapeutic.