The study confirms associations between epilepsy in Australian Shepherd dogs and two locations on chromosomes 1 (CFA1) and 19 (CFA19) found in previous research. PCR-RFLP tests on 88 additional dogs showed different genotype frequencies between normal and affected dogs, supporting genes in these locations that may cause epilepsy. Sequencing of the DOK6 gene near associated SNPs on CFA1 found no differences between one affected and unaffected dog. The results support the hypothesis that mutations in these chromosomal regions contribute to epilepsy in Australian Shepherds.
Mucolipidoses Tipo II e III e Gagueira Não-Sindrômica são condições genéticas...Stuttering Media
http://gagueira.wordpress.com
Mutações homozigóticas nos genes GNPTAB e GNPTG estão classicamente associadas às mucolipidoses tipo II (ML II, alfa/beta) e tipo III (ML III, alfa/beta/gama), que são doenças raras de armazenamento lisossômico caracterizadas por múltiplas patologias. Recentemente, variantes nos genes GNPTAB, GNPTG e NAGPA (este último funcionalmente relacionado aos dois primeiros) foram associadas com a gagueira persistente não-sindrômica. Analisando uma amostra de escala mundial que abrangia 1.013 indivíduos com gagueira persistente não-sindrômica, sem relação de parentesco, encontramos 164 indivíduos que carregavam alguma rara variante não-sinônima de codificação em um destes três genes. Comparamos a frequência destas variantes com aquelas encontradas em grupos-controle nas mesmas populações e também em bancos de dados genômicos, e confrontamos a localização dessas mutações com a localização das mutações relatadas nas mucolipidoses. Descobrimos que pessoas com gagueira exibiam um excesso de variantes não-sinônimas de codificação quando comparadas a pessoas dos grupos-controle e a genomas pertencentes às bases de dados dos projetos “1000 Genomes” e “Exome Sequencing”. Ao todo, foram identificadas 81 variantes diferentes nos casos de gagueira analisados pelo nosso estudo. Praticamente todas eram substituições do tipo missense, das quais apenas uma tinha sido previamente relatada na mucolipidose, uma doença geralmente associada a mutações mais amplas, que levam a uma perda completa de função da proteína codificada. Nossa conclusão é que essas raras variantes de codificação não-sinônimas nos genes GNPTAB, GNPTG, e NAGPA podem ser responsáveis por até 16% dos casos de gagueira persistente, e que as alterações nos genes GNPTAB e GNPTG ocorrem em posições da cadeia nucleotídica que, em geral, causam efeitos menos graves na função da proteína do que aqueles vistos nas mucolipidoses tipo II e III.
Dr. Paulo Arruda - Continuing Diagnostic Investigation–Novel SapelovirusJohn Blue
Continuing Diagnostic Investigation–Novel Sapelovirus - Dr. Paulo Arruda, from the 2016 Allen D. Leman Swine Conference, September 17-20, 2016, St. Paul, Minnesota, USA.
More presentations at http://www.swinecast.com/2016-leman-swine-conference-material
Activation of surrogate death receptor signalling triggers peroxynitrite depe...Saurabh Shekhar
includes information about cisplatin resistance cancer cells and their execution through peroxynitrite triggered apoptosis due to death signaling receptors basedon the findings of research article published in cell death and diseases.
Mucolipidoses Tipo II e III e Gagueira Não-Sindrômica são condições genéticas...Stuttering Media
http://gagueira.wordpress.com
Mutações homozigóticas nos genes GNPTAB e GNPTG estão classicamente associadas às mucolipidoses tipo II (ML II, alfa/beta) e tipo III (ML III, alfa/beta/gama), que são doenças raras de armazenamento lisossômico caracterizadas por múltiplas patologias. Recentemente, variantes nos genes GNPTAB, GNPTG e NAGPA (este último funcionalmente relacionado aos dois primeiros) foram associadas com a gagueira persistente não-sindrômica. Analisando uma amostra de escala mundial que abrangia 1.013 indivíduos com gagueira persistente não-sindrômica, sem relação de parentesco, encontramos 164 indivíduos que carregavam alguma rara variante não-sinônima de codificação em um destes três genes. Comparamos a frequência destas variantes com aquelas encontradas em grupos-controle nas mesmas populações e também em bancos de dados genômicos, e confrontamos a localização dessas mutações com a localização das mutações relatadas nas mucolipidoses. Descobrimos que pessoas com gagueira exibiam um excesso de variantes não-sinônimas de codificação quando comparadas a pessoas dos grupos-controle e a genomas pertencentes às bases de dados dos projetos “1000 Genomes” e “Exome Sequencing”. Ao todo, foram identificadas 81 variantes diferentes nos casos de gagueira analisados pelo nosso estudo. Praticamente todas eram substituições do tipo missense, das quais apenas uma tinha sido previamente relatada na mucolipidose, uma doença geralmente associada a mutações mais amplas, que levam a uma perda completa de função da proteína codificada. Nossa conclusão é que essas raras variantes de codificação não-sinônimas nos genes GNPTAB, GNPTG, e NAGPA podem ser responsáveis por até 16% dos casos de gagueira persistente, e que as alterações nos genes GNPTAB e GNPTG ocorrem em posições da cadeia nucleotídica que, em geral, causam efeitos menos graves na função da proteína do que aqueles vistos nas mucolipidoses tipo II e III.
Dr. Paulo Arruda - Continuing Diagnostic Investigation–Novel SapelovirusJohn Blue
Continuing Diagnostic Investigation–Novel Sapelovirus - Dr. Paulo Arruda, from the 2016 Allen D. Leman Swine Conference, September 17-20, 2016, St. Paul, Minnesota, USA.
More presentations at http://www.swinecast.com/2016-leman-swine-conference-material
Activation of surrogate death receptor signalling triggers peroxynitrite depe...Saurabh Shekhar
includes information about cisplatin resistance cancer cells and their execution through peroxynitrite triggered apoptosis due to death signaling receptors basedon the findings of research article published in cell death and diseases.
Как выжать максимум из контекстной рекламы в E-commerceOrigami
Из слайдов вы узнаете, как работать с контекстной рекламой в электронной торговле.
Презентация с конференции ECOM Expo 2016. Спикер – Елена Фирсова, директор по продукту Origami.
Выступление Елены Фирсовой на секции «Контекстная реклама – максимум performance. Тренды 2015 года».
Основные тезисы:
• Результаты тестов Origami наглядно показывают, что неэффективно расходуется от 20% до 50% рекламного бюджета. При этом нужно понимать, что это происходит каждый месяц, ведь рекламные кампании – это долгосрочная история. Поэтому важно искать и находить инструменты оптимизации конверсий (на рынке они есть).
• Существуют два метода измерения эффективности – последовательное тестирование и шахматное тестирование.
• В презентации демонстрируются преимущества и недостатки каждого метода, а также кейсы по тематике.
• Эффективные оптимизаторы конверсий существуют и есть рабочие методы оценки эффективности, благодаря которым возможно точно узнать сумму экономии бюджета на контекстную рекламу.
SNP discovery in African taurine and Zebu cattle by whole genome sequencing o...ILRI
Poster by Noyes HA, Agaba M, Anderson SI, Archibald AL, Ashelford K, Bradley D, Brass A, Finalyson HA, Hanotte O, Kay S, Kemp SJ, Khodadadi M, Law AS, Lu Z, Smith S, Talbot R, and Hall N. For the BecA Opening, Nairobi, 5 November 2010
A SNP array for human population genetics studiesAffymetrix
Yontao Lu, Teri Genschoreck, Swapan Mallick, Amy Ollmann, Nick Patterson, Yiping Zhan, Teresa Webster, David Reich Overview of the Human Origins Array, the first array developed with leading geneticists to enable rigorous population genetics studies.
Se han hecho públicos los resultados de un prometedor trabajo encabezado por investigadores del Hospital Infantil de Boston y la Facultad de Medicina de Harvard, que ha conseguido recuperar, utilizando terapia génica, parte de la audición de ratones sordos. El artículo, que ha merecido la portada de la prestigiosa revista Science Translational Medicine, promete abrir un abanico terapéutico para el tratamiento de la sordera genética en los seres humanos.
A Case Of Canine Trypanosomosis With Epistaxis In A TwoYear Old Alsatian Dogiosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
this is all of the information that I have please help Lab 5 In.pdfambikacomputer4301
this is all of the information that I have please help Lab 5 Introduction - Genetic mapping See
figure 5.1 for a schematic of the fly the cross you initially started with, you'll either crosses you
have been working on. Two labs ago, map the distance between the w gene and the m you set up
a pair of reciprocal parental crosses, gene, or between the w gene and the y gene. between mutant
and wild type flies (fig. 5.1a). You had one of two different mutant strains, each with two mutant
phenotypes - either white eyes all F2 individuals will receive only recessive (w) and miniature
wings (m), or white eyes (w) alleles from this parent (fig. 5.1d, orange and and yellow body (y).
The phenotypes of the F1 yellow chromosomes). Because of this, the flies should have indicated
to you that all mutant phenotype of each F2 fly will tell you which phenotypes in question are x-
linked recessive alleles (mutant or WT) were inherited from the (fig. 5.1b). heterozygous female
F1 parent (fig. 5.1d, dark and light blue chromosomes). The first F2 fly Last lab, you used the F1
flies from one of shown in figure 5.1d inherited 'a B' from the your parental crosses to set up an
F1 cross (fig. heterozygous parent and will end up with the consequence. After crossing two pure
breeding this F2, you observe a ABphenotype and parents, F1 offspring will be heterozygous for
therefore know that this fly received 'A B' from nearly all genes in question - the exception is X -
the heterozygous parent. linked genes in the male offspring. Since the Y chromosome is
equivalent to recessive alleles for The goal of genetic mapping is to X-linked genes, these F1
males are recessive for determine the likelihood of cross over between all X-linked genes and act
as a test cross. The two loci/genes. If we score the phenotypes of a heterozygous F1 females and
recessive F1 males large F2 population from our crosses, we can (test cross) can be used to map
the distance determine the recombination frequency of your between the genes causing the two
phenotypes two genes. of your parental mutant female. Depending on
e) F2 phenotype scoring: f) Recombination frequency: Eigure 5.1: Schematic of your Drosophila
crosses, See text of lab 5 intro for description.
For a given F1 gamete for the F2 individual it number of flies, it is easy to calculate the creates),
if no cross over occurs between the two recombinant frequency between your two genes genes in
question, the F2 phenotype will be the (fig. 5.1ef ). same as one of the original parents - either
fully WT or double mutant in this case. In figure 5.1d Recall from last time that a lower
recombinant these have blue chromosomes of a single colour. frequency is observed when
genetic map If a crossover does occur between the two genes, distances are small. When genes
are close to the F2 fly will have a phenotype unlike either of each other, there's a narrow range
on the the parents - a recombinant phenotype (shown chromosome for a random crossover to
land.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
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Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
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2 Case Reports of Gastric Ultrasound
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1. Epilepsy in Australian Shepherd Dogs
Valeria Rivera1,2, Katie M. Minor B.A3; Eva Furrow V.M.D., Edward (Ned)
Patterson D.V.M., Ph.D.3
1.University of Minnesota, Life Sciences Summer Undergraduate Research Program:
Heart, Lung, & Blood
2.University of Puerto Rico at Cayey
3. Department of Veterinary Clinical Sciences, University of Minnesota College of
Veterinary Medicine, St. Paul, MN.
Abstract
Idiopathic epilepsy implies having recurrent seizures in which no anomaly is
known. It is a common disorder that affects about 4% of dogs and 1% of people.
Previous Whole Genome Analysis showed statistical association of single nucleotide
polymorphism (SNP) with Epilepsy in a study of 40 Australian shepherd dogs
(Aussies) on loci of CFA1 and CFA19. This research further evaluated SNPs between
epileptic and non-epileptic Aussies.
The major objectives of this portion of the study were to confirm the statistical
associations observed on CFA1 and CFA19 in additional case and control Aussies via
PCR-RFLP tests and to sequence one candidate gene, DOK6. Two PCR-RFLP SNP
tests were performed on 88 additional Aussies to compare their genotype
frequencies by Chi-square statistical analysis. Sequencing of DOK6 was performed
on one case and one control Aussie.
By studying the SNPs in Aussies, we confirmed different genotype frequencies
between normal and affected dogs on canine chromosomes 1 and 19 indicating
possible causative genes in each location. Sequencing of DOK6 did not show any
polymorphisms in the exons (coding regions).
Key words: Epilepsy, canines, genetic basis, SNPs, and Australian Shepherds.
condition. Some studies suggest that ion
Introduction channel genes cause epilepsy for many of
the human forms. The known human
Epilepsy is a brain disorder of mutations have not shown to be the cause
abnormal electrical signals that cause in four dog breeds (Ekenstedt et al. 2011).
seizures. It varies in severity and is Breeds such as Bernese Mountain dogs,
common in children and adults. Vizslas, Labrador Retrievers, and
Idiopathic Epilepsy (IE) involves Australian Shepherds have a high
recurrent seizures in which no underlying incidence of epilepsy. For this study,
abnormality can be identified (Patterson Australian Shepherds were chosen
et al. 2003). Each year 200,000 new because of their high frequency of
human cases are diagnosed, being the epilepsy that appears to be inherited
second most common neurological based on pedigree analysis. As
1
2. generations pass, there are some genes in G is strongly associated with epilepsy.
close physical proximity on the same Chromosome 1 has one of the associated
chromosome that do not recombine and SNPs between C and T, with C being
are in what is called linkage strongly associated with epilepsy. The
disequilibrium (LD). In regions of LD SNP BICF2G63036912 on CFA19 is
there is a haplotype that is shared located at 13,956,701 bp of the
between closely related breeds and chromosome and the SNP
common ancestors. Long LD allows many BICF2G630711334 on CFA1 is located at
fewer genetic markers and cases to be 12,632,317 bps. On the canine SNP array,
used for genome wide association studies there is on average one SNP every
in dogs compared to people (Sutter et al. 14,000bp in the total of 2.4 billion bps of
2004). Breeds that have passed through the canine genome.
population bottlenecks are prone to carry DOK6 is a gene located in
more recessive traits since inbreeding is chromosome 1 that is very near some of
frequently employed. As a result, the associated SNP markers. It has 8
inbreeding may be responsible for coding exons. Our objective was to
common diseases in these breeds. perform Chi-square analysis for 88
Diseases include cancer, epilepsy, additional Australian Shepherds for two
deafness, and heart diseases. of the associated SNPs to determine if the
Whole genome association (WGA) original association was real or only by
is a technique that examines the genome chance. A second objective was to
of different individuals searching for determine if there are any coding
differences among them. DNA samples mutations in DOK6 for epilepsy in
are placed on a crystal chip that Australian Shepherds. The overall
genotypes 170,000 canine SNPs. A SNP is hypothesis of this work is that there is
a single nucleotide polymorphism, a one or more epilepsy causing mutations
difference in base pairs when a single in these specific regions of canine
nucleotide is altered at a specific location. chromosomes 1 and/or 19.
The lab team performed previous work
(WGA) for 40 Australian Shepherds in Figure 1: Aussies Chi-square test
which they established that there is a
possibility of one or two genes that cause
epilepsy in this breed.
These genes are likely to be at
specific locations on canine chromosomes
1 (CFA1) and/or CFA19, based on Chi-
square analysis of the data and the
resulting p-values. To obtain this data,
they tested 19 affected dogs and 21
unaffected. CFA1 and CFA19 showed a
number of SNPs close together that stand
out (Figure 1) when graphed on a Figure 1. Results of a WGA study with 19 dogs
negative logarithmic scale of their Chi- affected by epilepsy and 21 unaffected. Shows SNPs
square p-value (-log p-value). In that stand out when graphed on a negative
chromosome 19, one of the associated logarithmic scale of their Chi-square p-value (-log p-
SNPs is between nucleotides G and A, and value) on the y-axis and the canine chromosomes in
2
3. order on the x-axis. These are in CFA1 (pink) and in when the sequence T G A T C is present.
CFA19 (green). CFA19 has one that is statistically Where as BsmI (CFA19) recognizes
significant being over 1.3, equivalent to a p-value
less than 0.05. nucleotide G and will cut there every time
the sequence G C A T C is present. For
Materials and Methods CFA1 RFLP, 8 units (2µL) of sau3AI were
used along with NEB 1(2µL), BSA (0.2µL),
Dog DNA samples were collected and water (10.8µL). The 8 unit master
along with their medical information. mix (15µL) was pipetted into PCR
Affected dogs must have had their first product to digest overnight at 37°C hold.
seizure before 6 years, whereas normal For CFA19, 5 units of the enzyme BmsI
dogs were at least 8 years without a (0.5µL) were used leaving it digest for 3
history of seizures. In order for them to hours. The resulting banding patterns
be included in this study, they must have were visualized on a 2% agarose gel as
fulfilled these requirements: 1. Two or follows: 385bp for homozygous TT or AA;
more seizures 2. Normal in between of 385, 216 and 169 bp for heterozygous CT
seizures 3. Normal blood panels (no or AG, and 216bp and 169bp for
abnormalities). homozygous CC or GG for CFA1 and
Based on the WGA results, one dog CFA19 respectively. The RFLP results
of each genotype was selected to serve as determined the genotype of the additional
a control for the enzyme digest. For CFA 88 dogs’ samples leading the way for the
19, the genotypes were homozygous AA, statistical Chi-square analysis to verify
heterozygous AG, and homozygous GG. the association found by the prior WGA
For CFA1, the genotypes were study.
homozygous TT, heterozygous CT, and Additionally, DOK6 sequencing
homozygous CC. These 6 control samples was completed in one affected and one
served as references when determining unaffected dog. As described above, PCR
the genotypes of an additional 88 dogs via was done with primers that encompassed
RFLP. SNPs were chosen based on p-value all 8 exons. Sanger sequencing was then
and availability of RFLP. performed, and the results were
Fifteen ng of purified DNA, specific compared between the two dogs and
20 µM forward and reverse primers against the published dog sequence.
(1.5uL), 300 µM dNTPs (1.5µL), 10x
buffer (1.5µL) in water (5.6µL), and TAQ Figure 2: RFLP for CFA19
polymerase (0. 5 µL) were used for PCR to
amplify the region containing each SNP.
Cycling conditions were: 95°C for 15 min,
35 cycles of 95°C for 30 s, 60°C for 30 s,
and 72°C for 30 s; and 72°C for 15 min.
The PCR bands of 385 bp for both CFA1
and CFA19 were visualized on a 2%
agarose gel.
Enzymes Sau3AI (8u) and BsmI
(5u) were used for each of the 2
chromosomes of interests. Sau3AI (CFA1)
recognizes nucleotide C and cuts there Figure 2 Shows the 3 different band sizes after
3
4. performing the RFLP test for CFA19. The last three greater C allele frequency for affected dogs.
samples are the controls for homozygous AA • CFA19 showed statistical significance with
heterozygous AG and homozygous GG. For this a greater G allele frequency for affected
chromosome, G is strongly associated with dogs.
epilepsy; therefore, it is hypothesized that affected Conclusion
dogs have most often have a genotype of GG in the
studied position of 13,956,701bp.
Epilepsy in Australian Shepherds
is suspected to be an inherited disease
Results
and prior work has shown an association
between loci on CFA1 and CFA19 in this
Significant associations of SNPs
seizure disorder. By testing 23 affected
between affected and unaffected Aussies
and 65 unaffected Australian Shepherd
were still observed on CFA1 and CFA19
dogs for 2 different RFLP SNPs and
with the addition of (23) cases and (65)
comparing genotypes, the association of
controls, for a total of (42) cases and (86)
the loci on both CFA1 and CFA19 with
controls. The G allele in CFA19 (SNP -
epilepsy was confirmed. It is concluded
BICF2G63036912) is strongly associated
that there are likely genetic mutations in
with epilepsy and has a frequency of
both regions that contribute to the
47.3% in affected dogs. In CFA1 (SNP -
development of epilepsy in Australian
BICF2G630711334), the frequency for the
Shepherds.
C allele is also associated with epilepsy
Future work could involve WGA in
and has a frequency of 51.3% in affected
closely related breed such as Border
dogs.
Collies. Based on previous research on
The p-values for association for
Vizslas and English Springer Spaniels, the
these two SNP with epilepsy were 0.0103
lab team may continue their investigation
and 0.000750 for CFA1 and CFA19,
focusing on these 2 chromosomal areas. A
respectively. These results strongly
SNP Chi-square analysis could serve to
support our hypothesis of a causative
compare between breeds and determine
mutation near these locations in CFA1
if these associations are shared across
and CFA19. Affected dogs are more likely
breeds. In addition, new research may
to have G allele for the CFA19 SNP and the
involve looking at the other genes within
C allele for the CFA1 SNP as shown in
1-2 million base pairs near DOK6 to
Figure 3.
evaluate for a causative mutation for
Sanger sequence of 8 exons of DOK6
epilepsy in the breed.
did not reveal any differences in the case
versus the control Aussie selected.
Acknowledgements
I want to give special thanks to my
Figure: 3 Allele frequencies
mentor Dr. Ned Patterson and Katie
Minor for all their effort in guiding me
during my summer research project. Also,
I want to thank Dr. Eva Furrow for her
help with Chi-square analysis.
Funded by:
AKC Canine Health Foundation
Figure 3.
• CFA1 showed statistical significance with a LSSURP
4
5. RISE program R25GM059429-12
References
1. Ekenstedt k, Patterson E, Minor K et al.
2011. Candidate genes for idiopathic
epilepsy in four dog breeds BMC Genetics
[Internet]. Available from:
http://www.biomedcentral.com/1471-21
56/12/38
2. Patterson E, Mickelson J, Da Y et al.
2003. Clinical Characteristics and
Inheritance of Idiopathic Epilepsy in
Vizlas. Journal of Internal Veterinary
Medicine Volume 17, Issue 3, pages
319-325, May 2003. [Internet]. Available
from:
http://onlinelibrary.wiley.com/doi/10.11
11/j.1939-1676.2003.tb02455.x/abstract
3. Sutter N, Eberle M, Parker H et al.2004.
Extensive and breed-specific
linkage disequilibrium in Canis familiaris.
[Internet]. Available from:
http://www.ncbi.nlm.nih.gov/pmc/articl
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