Sperm nuclear integrity assessment [autosaved]
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This document summarizes various methods for assessing sperm nuclear integrity. It discusses the importance of sperm nuclear DNA for fertility and causes of abnormality. Several techniques are described that can detect DNA damage in sperm, including the Comet assay, TUNEL assay, SCSA test, and SCD test. The Comet assay measures DNA strand breaks by visualizing displacement of genetic material after lysis and electrophoresis. The TUNEL assay detects DNA fragmentation by labeling double strand breaks. The SCSA test indirectly evaluates DNA breaks through measuring acid-induced denaturation and staining with a fluorescent dye. These methods provide a way to evaluate sperm quality beyond standard semen analysis parameters.
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DNA fingerprinting is a method used to identify individuals based on unique characteristics in their DNA. Alec Jeffreys developed this technique in 1987 to solve criminal cases. DNA fingerprinting analyzes genetic diversity by using molecular markers like RFLP, AFLP, RAPD, SSRs, and ISSRs that can detect polymorphisms between individuals. One study analyzed genetic variation in 24 earthworms from different locations using RAPD and ISSR markers and found high levels of polymorphism. Another study examined genetic diversity in sheep from four flocks and found greater diversity between flocks than within flocks. So DNA fingerprinting is widely used to study genetic variation within and between populations.
Dnaprofiling
DNA profiling uses regions of non-coding DNA that vary between individuals, called short tandem repeats (STRs), to identify a person from their DNA. STR analysis has replaced older DNA fingerprinting techniques. It can analyze small amounts of DNA quickly and is used in forensics, parentage testing, and other applications. Parentage testing involves collecting samples from a child and putative parents, analyzing STR regions through PCR and electrophoresis, and using statistical analysis to calculate probability of paternity.
Forensic Anthropology الاستعراف
This document discusses forensic anthropology and some of its key methods. It describes forensic anthropology as the study of identifying, analyzing, and interpreting human skeletal remains from crime scenes. It outlines three main tasks of forensic anthropologists: identifying victims through biological profiles, examining evidence of trauma, and reconstructing the postmortem period. It also discusses specific methods like estimating age through skeletal and dental development, determining sex through physical traits, analyzing fingerprints and DNA.
DNA FINGERPRINTING SMG
DNA fingerprinting is a technique used to identify individuals by analyzing genetic variations in DNA sequences found in certain regions of the genome. It was developed in 1985 by Alec Jeffreys and involves isolating DNA from a sample, cutting it with restriction enzymes, and comparing patterns of DNA fragments between individuals using probes that bind to variable number tandem repeats. DNA fingerprinting relies on differences in repetitive sequences that vary between people but are identical in each cell of an individual, allowing a unique genetic profile to be generated for identification purposes in forensic investigations and paternity testing.
The power of dna fingerprinting in forensic science
This document discusses DNA fingerprinting, including what it is, how it works, and its uses. DNA fingerprinting is a forensic technique that identifies individuals by characteristics of their DNA using variable tandem repeats. It was invented in 1985 and involves extracting DNA from samples, amplifying it, separating fragments by size via electrophoresis, and comparing band patterns to identify matches. DNA fingerprinting is used for paternity testing, criminal investigations, medical identification, and other applications like solving wildlife crimes and identifying mass casualties. Famous cases that have been solved using this technique include proving Steve Bing's paternity and catching the serial killer Colin Pitchfork.
PCR (1).pptx
This document provides an overview of polymerase chain reaction (PCR), including:
- PCR was developed in 1983 by Kary Mullis to amplify specific DNA sequences.
- It works by using DNA polymerase to replicate short DNA fragments defined by primer binding sites, exponentially amplifying the target sequence through repeated heating and cooling cycles.
- Components include a DNA template, DNA polymerase, primers, nucleotides, and buffer solution.
- Common PCR techniques include real-time PCR, nested PCR, quantitative PCR, and arbitrary primed PCR.
- The major steps of PCR are denaturation, annealing of primers, and elongation through DNA polymerase activity.
- Applications include medicine, forensics, and paternity testing
Fish - Fluorescence In Situ Hybridization
This document provides an overview of fluorescence in situ hybridization (FISH). FISH is a cytogenetic technique used to detect and localize specific DNA sequences on chromosomes. The document discusses the history of FISH, basic methodology which involves using fluorescent probes and microscopy, sample and probe requirements, clinical applications for detecting cancers and genetic abnormalities, and factors that can affect FISH results. In summary, FISH is an important technique for detecting chromosomal abnormalities, it has applications in cancer diagnosis and genetic testing, and accurate performance requires consideration of many technical factors.
Fish - Fluorescence In Situ Hybridization
This document provides an overview of fluorescence in situ hybridization (FISH). FISH is a cytogenetic technique used to detect and localize specific DNA sequences on chromosomes. The document discusses the history of FISH, basic methodology, sample and probe requirements, clinical applications for detecting various cancers and genetic abnormalities, factors affecting FISH results, and recent advances like spectral karyotyping. In summary, FISH allows rapid detection of genetic changes through fluorescent labeling and hybridization of DNA probes to complementary sequences on chromosomes.
Dna fingerprinting by shalini mishra..
Madhumita was pregnant at the time of her murder. Her fetus was sent for DNA fingerprinting to determine paternity and identify the perpetrator. DNA fingerprinting works by extracting DNA from a sample, amplifying it if needed, cutting it with restriction enzymes, separating the fragments by size using gel electrophoresis, blotting it onto a medium, treating it with a radioactive probe, and creating an image on x-ray film to view the individual's unique DNA pattern. DNA fingerprinting has applications in determining paternity, identifying criminals, studying genetic variability in endangered species, and detecting pathogens or GMOs.
Forensic odontology
This document discusses forensic odontology and its role in personal identification. It begins by defining forensic odontology and its uses in legal investigations. Key aspects covered include methods of identifying unknown remains through dental records, determining ethnicity, gender, and age. Dental identification procedures for both single and mass casualty events are described. The document also discusses using dental DNA and palatal rugae patterns for identification when dental records are unavailable. In summary, this document provides an overview of forensic odontology and its applications in personal identification.
Dna fingerprinting
DNA fingerprinting is a technique developed in 1984 that analyzes variable regions in genetic material to distinguish one person from another. It involves isolating DNA from a sample, cutting it with restriction enzymes, sorting by size, and probing specific locations to generate a unique pattern. This technique uses variations in the number of short tandem repeats between individuals and has been used successfully in criminal cases and establishing paternity. The most common DNA fingerprinting methods are electrophoresis, polymerase chain reaction, restriction fragment length polymorphism, random amplified polymorphic DNA, and amplified fragment length polymorphism. DNA fingerprinting has applications in diagnosing inherited disorders, developing cures, and identifying criminals using biological evidence.
Single Nucleotide Polymorphism
This document discusses single nucleotide polymorphisms (SNPs), which are variations in DNA sequences among individuals. SNPs are the most common type of genetic variation, occurring around every 100 to 300 bases. They can be found in coding and non-coding regions, and may alter protein function directly by changing amino acids or indirectly by affecting regulatory sequences. SNPs can influence disease susceptibility and treatment response. Methods for identifying SNPs involve detecting known SNPs using techniques like restriction enzyme analysis or sequencing to find new SNPs. SNPs have many applications including gene mapping, disease association studies, diagnostics, and predicting treatment response.
Molecular Markers and Their Application in Animal Breed.pptx
Molecular markers have had a significant impact on breed development and conservation efforts, transforming genetics and offering vital insights into genetic diversity, lineage tracing, and genotype characterization. The importance of molecular markers in improving genetic gains, facilitating breeding programs, and preserving genetic diversity for the long-term sustainability of the animal population has been underlined in this review paper. Emerging advancements in molecular marker technology show enormous potential for improving and conserving breeds. Deeper insights into the genetic basis of complex traits will be provided through GWAS, CRISPR/Cas9, gene editing technologies, and sequencing technologies, resulting in faster genetic gains. Breeders and conservationists will be able to make more informed judgments thanks to these technologies. In conclusion, molecular markers have had a significant impact on breed conservation and enhancement. Their innovations have changed the industry and given both conservationists and breeders vital knowledge. We can pave the road for more effective and sustainable genetic improvement and the preservation of biodiversity for future generations by combining the power of molecular markers with conventional breeding and conservation techniques.
Application & ethical issues of gene sequencing
Its about application and ethical issues of gene sequemcing and its a simple one to understand by its point.
Similar to Sperm nuclear integrity assessment [autosaved] (20)
The power of dna fingerprinting in forensic science
The power of dna fingerprinting in forensic science
Molecular Markers and Their Application in Animal Breed.pptx
Molecular Markers and Their Application in Animal Breed.pptx
Recently uploaded
Immersive Learning That Works: Research Grounding and Paths Forward
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...Advanced-Concepts-Team
Presentation in the Science Coffee of the Advanced Concepts Team of the European Space Agency on the 07.06.2024.
Speaker: Diego Blas (IFAE/ICREA)
Title: Gravitational wave detection with orbital motion of Moon and artificial
Abstract:
In this talk I will describe some recent ideas to find gravitational waves from supermassive black holes or of primordial origin by studying their secular effect on the orbital motion of the Moon or satellites that are laser ranged.Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
The binding of cosmological structures by massless topological defects
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...
The pygmy halfbeak Dermogenys colletei, is known for its viviparous nature, this presents an intriguing case of relatively low fecundity, raising questions about potential compensatory reproductive strategies employed by this species. Our study delves into the examination of fecundity and the Gonadosomatic Index (GSI) in the Pygmy Halfbeak, D. colletei (Meisner, 2001), an intriguing viviparous fish indigenous to Sarawak, Borneo. We hypothesize that the Pygmy halfbeak, D. colletei, may exhibit unique reproductive adaptations to offset its low fecundity, thus enhancing its survival and fitness. To address this, we conducted a comprehensive study utilizing 28 mature female specimens of D. colletei, carefully measuring fecundity and GSI to shed light on the reproductive adaptations of this species. Our findings reveal that D. colletei indeed exhibits low fecundity, with a mean of 16.76 ± 2.01, and a mean GSI of 12.83 ± 1.27, providing crucial insights into the reproductive mechanisms at play in this species. These results underscore the existence of unique reproductive strategies in D. colletei, enabling its adaptation and persistence in Borneo's diverse aquatic ecosystems, and call for further ecological research to elucidate these mechanisms. This study lends to a better understanding of viviparous fish in Borneo and contributes to the broader field of aquatic ecology, enhancing our knowledge of species adaptations to unique ecological challenges.
The debris of the ‘last major merger’ is dynamically young
The Milky Way’s (MW) inner stellar halo contains an [Fe/H]-rich component with highly eccentric orbits, often referred to as the
‘last major merger.’ Hypotheses for the origin of this component include Gaia-Sausage/Enceladus (GSE), where the progenitor
collided with the MW proto-disc 8–11 Gyr ago, and the Virgo Radial Merger (VRM), where the progenitor collided with the
MW disc within the last 3 Gyr. These two scenarios make different predictions about observable structure in local phase space,
because the morphology of debris depends on how long it has had to phase mix. The recently identified phase-space folds in Gaia
DR3 have positive caustic velocities, making them fundamentally different than the phase-mixed chevrons found in simulations
at late times. Roughly 20 per cent of the stars in the prograde local stellar halo are associated with the observed caustics. Based
on a simple phase-mixing model, the observed number of caustics are consistent with a merger that occurred 1–2 Gyr ago.
We also compare the observed phase-space distribution to FIRE-2 Latte simulations of GSE-like mergers, using a quantitative
measurement of phase mixing (2D causticality). The observed local phase-space distribution best matches the simulated data
1–2 Gyr after collision, and certainly not later than 3 Gyr. This is further evidence that the progenitor of the ‘last major merger’
did not collide with the MW proto-disc at early times, as is thought for the GSE, but instead collided with the MW disc within
the last few Gyr, consistent with the body of work surrounding the VRM.
Equivariant neural networks and representation theory
Or: Beyond linear.
Abstract: Equivariant neural networks are neural networks that incorporate symmetries. The nonlinear activation functions in these networks result in interesting nonlinear equivariant maps between simple representations, and motivate the key player of this talk: piecewise linear representation theory.
Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptx
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
Bob Reedy - Nitrate in Texas Groundwater.pdf
Presented at June 6-7 Texas Alliance of Groundwater Districts Business Meeting
在线办理(salfor毕业证书)索尔福德大学毕业证毕业完成信一模一样
学校原件一模一样【微信：741003700 】《(salfor毕业证书)索尔福德大学毕业证》【微信：741003700 】学位证，留信认证（真实可查，永久存档）原件一模一样纸张工艺/offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原。
1:1完美还原海外各大学毕业材料上的工艺：水印，阴影底纹，钢印LOGO烫金烫银，LOGO烫金烫银复合重叠。文字图案浮雕、激光镭射、紫外荧光、温感、复印防伪等防伪工艺。材料咨询办理、认证咨询办理请加学历顾问Q/微741003700
【主营项目】
一.毕业证【q微741003700】成绩单、使馆认证、教育部认证、雅思托福成绩单、学生卡等！
二.真实使馆公证(即留学回国人员证明,不成功不收费)
三.真实教育部学历学位认证（教育部存档！教育部留服网站永久可查）
四.办理各国各大学文凭(一对一专业服务,可全程监控跟踪进度)
如果您处于以下几种情况：
◇在校期间，因各种原因未能顺利毕业……拿不到官方毕业证【q/微741003700】
◇面对父母的压力，希望尽快拿到；
◇不清楚认证流程以及材料该如何准备；
◇回国时间很长，忘记办理；
◇回国马上就要找工作，办给用人单位看；
◇企事业单位必须要求办理的
◇需要报考公务员、购买免税车、落转户口
◇申请留学生创业基金
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
20240520 Planning a Circuit Simulator in JavaScript.pptx
Evaporation step counter work. I have done a physical experiment.
(Work in progress.)
The cost of acquiring information by natural selection
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...Abdul Wali Khan University Mardan,kP,Pakistan
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skillsRandomised Optimisation Algorithms in DAPHNE
Slides from talk:
Aleš Zamuda: Randomised Optimisation Algorithms in DAPHNE .
Austrian-Slovenian HPC Meeting 2024 – ASHPC24, Seeblickhotel Grundlsee in Austria, 10–13 June 2024
https://ashpc.eu/
8.Isolation of pure cultures and preservation of cultures.pdf
Isolation of pure culture, its various method.
快速办理(UAM毕业证书)马德里自治大学毕业证学位证一模一样
学校原件一模一样【微信：741003700 】《(UAM毕业证书)马德里自治大学毕业证学位证》【微信：741003700 】学位证，留信认证（真实可查，永久存档）原件一模一样纸张工艺/offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原。
1:1完美还原海外各大学毕业材料上的工艺：水印，阴影底纹，钢印LOGO烫金烫银，LOGO烫金烫银复合重叠。文字图案浮雕、激光镭射、紫外荧光、温感、复印防伪等防伪工艺。材料咨询办理、认证咨询办理请加学历顾问Q/微741003700
【主营项目】
一.毕业证【q微741003700】成绩单、使馆认证、教育部认证、雅思托福成绩单、学生卡等！
二.真实使馆公证(即留学回国人员证明,不成功不收费)
三.真实教育部学历学位认证（教育部存档！教育部留服网站永久可查）
四.办理各国各大学文凭(一对一专业服务,可全程监控跟踪进度)
如果您处于以下几种情况：
◇在校期间，因各种原因未能顺利毕业……拿不到官方毕业证【q/微741003700】
◇面对父母的压力，希望尽快拿到；
◇不清楚认证流程以及材料该如何准备；
◇回国时间很长，忘记办理；
◇回国马上就要找工作，办给用人单位看；
◇企事业单位必须要求办理的
◇需要报考公务员、购买免税车、落转户口
◇申请留学生创业基金
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
Recently uploaded (20)
Immersive Learning That Works: Research Grounding and Paths Forward
Immersive Learning That Works: Research Grounding and Paths Forward
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...
aziz sancar nobel prize winner: from mardin to nobel
aziz sancar nobel prize winner: from mardin to nobel
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
The binding of cosmological structures by massless topological defects
The binding of cosmological structures by massless topological defects
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...
The debris of the ‘last major merger’ is dynamically young
The debris of the ‘last major merger’ is dynamically young
Equivariant neural networks and representation theory
Equivariant neural networks and representation theory
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptx
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptx
20240520 Planning a Circuit Simulator in JavaScript.pptx
20240520 Planning a Circuit Simulator in JavaScript.pptx
The cost of acquiring information by natural selection
The cost of acquiring information by natural selection
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
8.Isolation of pure cultures and preservation of cultures.pdf
8.Isolation of pure cultures and preservation of cultures.pdf
Sperm nuclear integrity assessment [autosaved]
- 2. OBJECTIVE OF STUDY SPERM NUCLEAR INTEGIITY ASSESSMENT • IMPORTANCE OF NUCLEAR DNA IN SPERM • CAUSE OF ABNORMALITY IN SPERM NUCLAR DNA • ASSESSMENT METHOD TO DETACT THE ABNORMALITY
- 3. IMPORTANCE OF NUCLEAR DNA IN SPERM • IN MOST OF THE WESTERN COUNTRIES AND INDIA ONE FOURTH COUPLES IN REPRODUCTIVE AGE ARE SEEKING MEDICAL HELP FOR INVOLUNTARY CHILDLESSNESS. • BOTH MALE GAMET AND FEAMALE GAMET HAD EQUAL WATAGE FOR A PERFECT DEVELOPMENT OF EMBRYO AND ITS FATE. • WHEN WE CAME TO THE PERSPECTIVE MALE, IN 1994 WHO DEVELOP STANDARD SEMEN PARAMETER ASSAY WHEATHER A MALE CONSIDER TO BE FERTILE OR INFERTILE. • BESIDE THIS PARAMETER SPERM DNA QUALITY IS IMPORTANT IN MAINTAINING THE REPROTIVE POTENTIAL OF MEN.
- 4. • THE FERTILITY POTENTIAL OF SPERM DEPENDS NOT ONLY UPON THE FUNCTIONAL COMPENTANCE OF SPERMATOZOA BUT ALSO ON SPERM DNA INTIGRITY. • MEN WITH NORMAL SPERMATOZOA MAY STILL INFERTILE THE CAUSE COULD BE RELATED TO ABNORMAL SPERM DNA. • IF SOME HOW THE FERTILIZATION OCCURE THE ZYGOTE WILL NOT DELVELOP NORMALLY. • THUS THE NUCLEAR SPERM INTIGRITY ASSESSEMNT METHODS IS DONE TO DETACT THE ABNORMALITY OF SPERM AT NUCLEAR AND GENETIC LEVEL.
- 5. CAUSE OF ABNORMALITY IN SPERM NUCLAR DNA
- 6. ASSESSMENT METHOD TO DETACT THE ABNORMALITY • MANY NEW TECHNIQUES HAVE BEEN DEVELOPED DURING THE PAST TWO DECADES TO ASSESS THE SPERM NUCLEAR INTEGRITY. • SUCH TECHNIQUES, USING BOTH MICROSCOPY- AND FLOW CYTOMETRY-BASED ANALYSES, EVALUATE SPERM DNA INTEGRITY AND SPERM CHROMATIN QUALITY. • CURRENTLY, FOUR MAJOR TESTS OF SPERM DNA FRAGMENTATION ARE MOST FREQUENTLY USED, INCLUDING THE COMET ASSAY (SINGLE CELL GEL ELECTROPHORESIS) , THE TUNEL (TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE- MEDIATED DUDP NICK END LABELLING) ASSAY , THE SPERM CHROMATIN DISPERSION (SCD) TEST AND THE SPERM CHROMATIN STRUCTURE ASSAY (SCSA). • THEY ALL LABEL SINGLE- OR DOUBLE-STRANDED DNA BREAKS; HOWEVER UNFORTUNATELY, MOST OF THE AVAILABLE TECHNIQUES FOR DETECTION OF SPERM DNA DAMAGE PROVIDE LIMITED INFORMATION ON THE NATURE OF THE DNA LESIONS DETECTED AND NONE OF THEM ENABLES US TO DEPICT THE EXACT AETIOLOGY AND
- 7. COMET ASSAY • THE COMET ASSAY IS A SIMPLE METHOD FOR MEASURING DNA STRAND BREAKS. • IN COMET ASSAY SPERM CELLS ARE MIXED WITH MELTED AGAROSE AND THEN PLACED ON A GLASS SLIDE. THE CELLS ARE LYSED AND THEN SUBJECTED TO HORIZONTAL ELECTROPHORESIS. • DNA IS VISUALIZED WITH THE HELP OF A DNA SPECIFIC FLUORESCENT DYE AND DNA DAMAGE IS QUANTIFIED BY MEASURING THE DISPLACEMENT BETWEEN THE GENETIC MATERIAL OF THE NUCLEUS COMET HEAD AND THE RESULTING TAIL.
- 8. TUNEL ASSAY • TUNEL ASSAY IS A METHOD FOR DETECTING DNA FRAGMENTATION BY LABELING THE 3’-HYDROXYL TERMINAL IN THE DOUBLE STRAND DNA BREAKS GENERATE DURING APOPTOSIS. • THE FLUORESCENCE INTENSITY OF EACH ANALYZED SPERM IS DETERMINED AS A “POSITIVE” OR “NEGATIVE” FOR SPERM ON A MICROSCOPE SLIDE.
- 9. SCSA TEST • SCSA IS A FLOW CYTOMETRY TEST WHERE SPERM DNA BREAKS ARE EVALUATED INDIRECTLY THROUGH THE DNA DENATURABILITY. THE ASSAY MEASURES THE SUSCEPTIBILITY OF SPERM DNA TO ACID INDUCED DNA DENATURATION IN SITU, FOLLOWED BY STAINING WITH THE FLUORESCENCE DYE ACRIDINE ORANGE. • BY USING A FLOW CYTOMETER 5.000–10.000 SPERM CAN BE ANALYZED WITHIN FEW SECONDS AND THUS PROVIDE A LESS SUBJECTIVE RESULT COMPARED TO THE WHO ANALYSIS WHERE ONLY 1–300 CELLS ARE ANALYZED. • THROUGH A SPECIFIC SCSA-SOFTWARE (SCSA- SOFT) A SCATTER PLOT IS CREATED, SHOWING THE RATIO OF GREEN AND RED SPERM. THE PERCENTAGE OF RED SPERM IS CALLED DNA
- 10. SCD TEST
- 11. THANK YOU