This document summarizes a research project investigating genetic variation in the CXCL16 gene among members of the Perissodactyla order. The project found that while horses share a susceptible allele, only horses have a resistant allele. Rhinocerids have a distinct amino acid sequence, indicating different selective pressure. While equids share the susceptible allele, the resistant allele appears unique to horses. This suggests the resistant allele is newly evolved in horses. More research is needed, including studying tapir sequences and obtaining more samples across species.
This study identified and characterized a new collagenase, ColAh, produced by Aeromonas piscicola AH-3. The researchers identified a gene in A. hydrophila ATCC 7966 that encodes a putative collagenase. They constructed a knockout mutant of this gene (colAh) in A. piscicola AH-3. Tests showed the mutant had significantly lower collagenolytic activity and cytotoxic effects compared to the wild type, demonstrating the colAh gene encodes an active collagenase. Further analysis found ColAh is a ~100kDa metalloprotease that can cleave and bind collagen. This contributes to A. piscicola's collagen degradation and cytotoxicity. ColAh contains the conserved HEXX
1) Cells expressing insulin and porcine proinsulin mRNA were found to engraft in the kidneys of diabetic rhesus macaques that were first transplanted with pig pancreatic primordia and later implanted with porcine islets, but no engraftment occurred without the initial primordia transplant.
2) Engrafted cells in the kidney and lymph nodes stained positive for insulin and porcine proinsulin mRNA. Fluorescent in situ hybridization detected pig X chromosomes in engrafted cells.
3) RT-PCR detected porcine proinsulin mRNA in the kidney of macaques receiving both transplants but not in macaques receiving only islet implants.
This document reports on the molecular characterization of male and female rat liver aldehyde oxidase (AOX). It finds that male and female rat livers express kinetically distinct forms of AOX, similar to what was observed in mice. However, sequencing of cDNAs from male and female rat liver suggests that the differences are likely due to differences in redox state rather than expression of separate genes, as only a single AOX gene appears to be expressed. Purification of the enzymes from each sex revealed a single 150 kDa protein, consistent with expression of a single gene. The kinetic parameters of the enzymes could be interconverted through chemical manipulation of redox state.
Population Genetic Models of Genomic ImprintingGavin Pearce
This document presents several population genetic models of genomic imprinting and draws the following conclusions:
(1) Systems with genomic imprinting do not necessarily behave the same as identical systems without imprinting.
(2) However, many of the models investigated can be shown to be formally equivalent to models without imprinting.
(3) Consequently, imprinting often cannot be discovered by following allele frequency changes or examining equilibrium values.
The document then goes on to describe four specific models of genomic imprinting and the population genetic dynamics that result from each.
- Drosophila melanogaster was one of the first organisms studied genetically due to its small size, short life cycle, high reproduction rate, and many genetic variants.
- Crosses were set up between red-eyed and white-eyed Drosophila strains to study inheritance patterns. The F1 and F2 progeny were examined to determine phenotypes.
- Most crosses showed Mendelian inheritance ratios, but some showed deviations, suggesting asymmetric or sex-linked inheritance patterns. Further test crosses were conducted to understand the genetics.
Study of psymberin's mode of action using forward geneticsVincent Tsao
Through forward genetics screening in C. elegans, the researchers identified a mutation in the rpl-41 gene that confers resistance to psymberin, a potent cytotoxin. Using single nucleotide polymorphism (SNP) mapping, they narrowed the location of the mutation to a region between +3.1 cM and +3.5 cM on chromosome II. Sequencing of the mutant gene identified a point mutation that results in an amino acid change from proline to leucine. This mutation in the conserved rpl-41 gene, which encodes a 60S ribosomal protein, suggests psymberin's mechanism of inducing cell death may involve additional pathways beyond just translation inhibition.
This document discusses estimating the number of sex alleles and queen matings in a population of Apis mellifera (honey bees) based on the frequency of diploid males observed. The number of sex alleles is estimated to be 18.9 based on data from a population of 90 hives in Brazil, which is larger than previous estimates of 10-12 alleles. The best estimate of the number of matings per queen is 17.3, obtained using a maximum likelihood approach assuming a prior distribution on the number of matings. The distribution of diploid males depends on both the number of sex alleles and the number of drones that mate with each queen.
This study identified and characterized a new collagenase, ColAh, produced by Aeromonas piscicola AH-3. The researchers identified a gene in A. hydrophila ATCC 7966 that encodes a putative collagenase. They constructed a knockout mutant of this gene (colAh) in A. piscicola AH-3. Tests showed the mutant had significantly lower collagenolytic activity and cytotoxic effects compared to the wild type, demonstrating the colAh gene encodes an active collagenase. Further analysis found ColAh is a ~100kDa metalloprotease that can cleave and bind collagen. This contributes to A. piscicola's collagen degradation and cytotoxicity. ColAh contains the conserved HEXX
1) Cells expressing insulin and porcine proinsulin mRNA were found to engraft in the kidneys of diabetic rhesus macaques that were first transplanted with pig pancreatic primordia and later implanted with porcine islets, but no engraftment occurred without the initial primordia transplant.
2) Engrafted cells in the kidney and lymph nodes stained positive for insulin and porcine proinsulin mRNA. Fluorescent in situ hybridization detected pig X chromosomes in engrafted cells.
3) RT-PCR detected porcine proinsulin mRNA in the kidney of macaques receiving both transplants but not in macaques receiving only islet implants.
This document reports on the molecular characterization of male and female rat liver aldehyde oxidase (AOX). It finds that male and female rat livers express kinetically distinct forms of AOX, similar to what was observed in mice. However, sequencing of cDNAs from male and female rat liver suggests that the differences are likely due to differences in redox state rather than expression of separate genes, as only a single AOX gene appears to be expressed. Purification of the enzymes from each sex revealed a single 150 kDa protein, consistent with expression of a single gene. The kinetic parameters of the enzymes could be interconverted through chemical manipulation of redox state.
Population Genetic Models of Genomic ImprintingGavin Pearce
This document presents several population genetic models of genomic imprinting and draws the following conclusions:
(1) Systems with genomic imprinting do not necessarily behave the same as identical systems without imprinting.
(2) However, many of the models investigated can be shown to be formally equivalent to models without imprinting.
(3) Consequently, imprinting often cannot be discovered by following allele frequency changes or examining equilibrium values.
The document then goes on to describe four specific models of genomic imprinting and the population genetic dynamics that result from each.
- Drosophila melanogaster was one of the first organisms studied genetically due to its small size, short life cycle, high reproduction rate, and many genetic variants.
- Crosses were set up between red-eyed and white-eyed Drosophila strains to study inheritance patterns. The F1 and F2 progeny were examined to determine phenotypes.
- Most crosses showed Mendelian inheritance ratios, but some showed deviations, suggesting asymmetric or sex-linked inheritance patterns. Further test crosses were conducted to understand the genetics.
Study of psymberin's mode of action using forward geneticsVincent Tsao
Through forward genetics screening in C. elegans, the researchers identified a mutation in the rpl-41 gene that confers resistance to psymberin, a potent cytotoxin. Using single nucleotide polymorphism (SNP) mapping, they narrowed the location of the mutation to a region between +3.1 cM and +3.5 cM on chromosome II. Sequencing of the mutant gene identified a point mutation that results in an amino acid change from proline to leucine. This mutation in the conserved rpl-41 gene, which encodes a 60S ribosomal protein, suggests psymberin's mechanism of inducing cell death may involve additional pathways beyond just translation inhibition.
This document discusses estimating the number of sex alleles and queen matings in a population of Apis mellifera (honey bees) based on the frequency of diploid males observed. The number of sex alleles is estimated to be 18.9 based on data from a population of 90 hives in Brazil, which is larger than previous estimates of 10-12 alleles. The best estimate of the number of matings per queen is 17.3, obtained using a maximum likelihood approach assuming a prior distribution on the number of matings. The distribution of diploid males depends on both the number of sex alleles and the number of drones that mate with each queen.
This document summarizes an experiment on Mendelian genetics involving monohybrid and dihybrid crosses. It describes using Punnett squares to determine genotypic and phenotypic ratios for crosses involving one or two traits. Chi-square tests were used to analyze if the observed ratios fit the expected Mendelian ratios. For monohybrid crosses, the observed ratios fit 1:2:1 genotypic and 3:1 phenotypic ratios. However, for some dihybrid cross samples, the observed ratios did not fit the expected 9:3:3:1 phenotypic ratio, possibly due to limitations in the experimental design and samples used.
This document presents information on complementation tests. It defines complementation tests as a method used to determine if two mutations are in the same gene or different genes. It explains that if the mutations are complementary (in different genes), the offspring will show the parental phenotypes, but if they are not complementary (in the same gene), the offspring will show a new phenotype. Three examples of using complementation test results to determine the number of genes involved are provided. The document concludes by citing a reference for more information on assigning mutations to genes using complementation tests.
Mendel performed a dihybrid cross experiment involving two traits - plant height and flower color. He crossed a tall, red-flowered homozygous plant with a dwarf, white-flowered homozygous plant. The F1 generation were all tall with red flowers, showing dominance. In the F2 generation, there were 9 tall red plants, 3 tall white plants, 3 dwarf red plants, and 1 dwarf white plant, in a 9:3:3:1 ratio. This supported Mendel's second law of independent assortment, that two traits are inherited independently. Test crosses on the F1 hybrids confirmed independent assortment of alleles between the two gene pairs.
This document describes the discovery and characterization of a common inversion polymorphism on chromosome 8p in humans. Analysis of recombination patterns in families identified apparent triple recombinations in a 12 cM region on chromosome 8p that were resolved by inverting the order of two markers. Fluorescent in situ hybridization confirmed the inversion in these families and others, estimating the inversion frequency at 21% in individuals of European ancestry. The inversion spans approximately 12 cM genetically and 2.5-5.3 Mb physically, and is flanked by clusters of olfactory receptor genes, suggesting it may be mediated by recombination between these repeats. The polymorphism could impact susceptibility to certain chromosomal abnormalities and influence gene expression near the breakpoint.
This document summarizes the discovery of a novel 162 base pair exon insertion, denoted exon 9a, in the interleukin 23 (IL-23) receptor gene. The insertion was found between exons 9 and 10 of the IL-23 receptor mRNA. Sequence analysis revealed splice donor-acceptor sites flanking the insertion, suggesting it is a legitimate cryptic exon. Although the insertion is in-frame, it contains a premature stop codon that would result in a truncated and non-functional IL-23 receptor protein. Further analysis detected the exon 9a insertion at low levels in mRNA from mitogen-stimulated immune cells, indicating it arises from rare alternative splicing of the IL-23 receptor pre-mRNA. Homologous
Poster: Effects of LEMS on P/Q type Calcium Channelsmaryvi
This study examined the effects of Lambert-Eaton myasthenic syndrome (LEMS) plasma on calcium channel expression in tottering (tg) mice, which have a mutation in the P/Q-type calcium channel subunit α1A. Western blots were used to analyze the expression of calcium channel subunits α1A, α1B, α1C, and α1E in muscle tissue from tg and wild-type mice treated with either LEMS plasma or saline solution. The results suggest that N-type and R-type calcium channels may compensate for the loss of P/Q-type channels in tg mice treated with LEMS plasma. However, the antibodies used showed low specificity, so more experiments
This document summarizes a study that identified 24 splice variants of the human IL-23 receptor (IL-23R) gene from activated human leukocytes. The variants were characterized through restriction enzyme digestion and sequencing of IL-23R cDNA clones. Most variants involved exon skipping or deletions in the extracellular region of the IL-23Ra protein, which could impact IL-23 ligand binding and signaling. The identification of these potentially functional IL-23R variants improves understanding of how the gene contributes to both normal immune function and pathological conditions like inflammatory bowel disease.
Thant Bio Symposium Poster Spring 2016Claire Thant
The study found that axonal transport defects activate the PI3K pathway in an attempt to reduce oxidative stress and neuronal cell death in neurodegenerative disease models. Expressing constitutively active PI3K decreased cell death caused by polyQ repeats and Paraquat exposure but did not affect axonal transport blockages. Dominant negative PI3K disrupted normal huntingtin motility, indicating PI3K acts downstream of transport. Motor protein mutations and disease models showed increased levels of p-GSK3β, a PI3K effector, suggesting transport defects trigger the protective PI3K response.
This study aimed to confirm previous findings linking canine chromosomes 1 and 19 to idiopathic epilepsy in Australian Shepherd dogs. PCR-RFLP tests of two SNPs were performed on 88 additional Australian Shepherds and genotypes were compared between affected and unaffected dogs via Chi-square analysis. Sequencing of the DOK6 gene did not reveal differences between one affected and unaffected dog. The results confirmed different genotype frequencies between normal and affected dogs for the SNPs, supporting the hypothesis that epilepsy-causing genes are located near the tested regions on chromosomes 1 and 19.
1) The study investigated the expression of the protein TMEM35 in different cell lines and its effects on surface expression of the nicotinic acetylcholine receptor α7 (α7 nAChR).
2) Western blot analysis showed that TMEM35 expression levels correlated with the ability of cell lines to express surface α7 nAChR after transfection.
3) Preliminary results suggest C-terminally tagged TMEM35 may not enhance α7 nAChR surface expression in the same way as untagged TMEM35, possibly due to interaction of the C-terminus with the chaperone protein RIC3.
This study investigated the role of histone H3 lysine 18 acetylation (H3K18ac) in maintaining pluripotency in embryonic stem cells. The researchers found that H3K18ac is highly enriched at stem cell genes in embryonic stem cells and levels decrease with differentiation. Expression of the E1A oncoprotein in mouse embryonic stem cells globally reduced H3K18ac and caused the cells to lose pluripotent characteristics, fail to activate stem cell enhancers, and suppress lineage-specific genes. Loss of pluripotency upon E1A expression required binding of E1A to the H3K18 acetyltransferases p300/CBP. This suggests H3K
antibody structure classification and functionsDUSHYANT KUMAR
Antibodies (immunoglobulins) are globulin proteins formed in response to antigens that specifically react with and destroy antigens. They are produced by plasma cells and found in blood serum, body fluids, and tissues. Antibodies are the immune system's way of targeting harmful foreign substances like bacteria, viruses, and more. Through experiments in 1939 and 1962, scientists discovered that antibodies are contained within the gamma globulin fraction of blood and have a basic four-chain structure. There are five classes of antibodies in human serum named for the type of heavy chain they contain: IgG, IgA, IgM, IgE, and IgD.
Molecular characterization of anaplasma platys strains from dogs in sicily, i...Josephine Huang
1) Researchers analyzed blood samples from 344 dogs in Sicily, Italy to characterize strains of Anaplasma platys.
2) They found A. platys DNA in 14 dogs (4% prevalence) through PCR and gene sequencing of the 16S rDNA, groESL, and gltA genes.
3) Sequence analysis identified at least 3 different genotypes of A. platys among the Sicilian dog samples based on variations in the gltA gene.
This document summarizes a study on the phylogeny of predatory stink bugs in the subfamily Asopinae. The analysis recovered the monophyly of Asopinae, which was supported by 16 synapomorphies. Eighteen of the 19 genera included in the analysis with multiple species were monophyletic, while the genus Podisus was paraphyletic. The pseudoclaspers found in males may have contributed to the evolutionary success of Asopinae by enhancing copulatory performance compared to other pentatomids.
Genetic polymorphism of pigs (Sus Scrofa domestica) of Ukrainian meat and Wel...Maria Drahulian
Genetic polymorphism of pigs (Sus Scrofa domestica) of Ukrainian meat and Welsh breeds according to cyto- and molecular- genetic markers /M. Drahulian, S. Kostenko, T. Dorosch // Cytopathology. Volume 29, Issue S1 Abstracts of the 41st European Congress of Cytopathology, Madrid, Spain, 10 – 13 June 2018. P - 129. DOI: 10.1111/cyt.12569
Elucidating changes in gene expression in Tryp susceptible and resistant cattle during progression of tryp infection using Affymetrix gene expression Micro arrays
Detection of ehrlichia platys dna in brown dog ticksJosephine Huang
This document reports on a study that detected Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) collected from dogs in Okinawa Island, Japan. Key findings include:
- Partial 16S rRNA gene sequences of E. platys were detected in 3 out of 32 ticks collected from 2 dogs using PCR and sequencing.
- This represents the first detection of E. platys in Japan and the first report of detecting it in ticks.
- Sequence analysis showed the 3 positive ticks were highly similar (99.7-100% identical) to known E. platys sequences, confirming the detection.
- This suggests R. sanguineus ticks
The document discusses a simulation of a population of bunnies in England, where some bunnies have a recessive allele that causes them to lack fur. Over generations, as the harsh winters cause the hairless bunnies to die off at higher rates, the frequency of the recessive allele decreases. The results demonstrate natural selection, as the recessive allele becomes less common as it confers a lower chance of survival in that environment.
This study aims to determine if differences in alleles of the APETALA1 (AP1) gene are responsible for phenotypic differences between varieties of Brassica oleracea, specifically cauliflower and Rbo. The researchers generated F1 and F2 crosses between cauliflower and Rbo, which showed segregating phenotypes. They are determining the genotypes of the AP1a and AP1c alleles in the F2 plants to test if there is a correlation between genotype and phenotype. Preliminary results found the AP1c sequence from cauliflower is identical to sequences from broccoli and kale, suggesting AP1 may not be responsible for phenotypic differences as hypothesized.
This study compared the contractile response patterns of ergot alkaloids isolated from tall fescue in bovine lateral saphenous veins. The ergoline alkaloids lysergic acid, lysergol, and ergonovine produced quicker contractile responses that began relaxing immediately, while the ergopeptine alkaloids ergovaline, ergotamine, ergocristine, ergocryptine, and ergocornine had slower, more persistent contractile responses with little to no relaxation over 120 minutes. The different classes of alkaloids produced significantly different contractile response patterns in the veins. Persistent vasoconstriction by certain ergot alkaloids may
This document discusses cattle losses due to larkspur poisoning on grazing lands in Utah. Typical cattle losses are around 5% but can reach 15%. Clinical signs of poisoning include staggering, muscle tremors, and collapse. Grazing management strategies recommend grazing early or late to avoid the "toxic window" when larkspur is most dangerous. A study evaluated cattle breeds' susceptibility and identified genetic markers associated with resistance in Angus cattle. Further research aims to validate these markers and identify other genes influencing larkspur resistance.
This document summarizes an experiment on Mendelian genetics involving monohybrid and dihybrid crosses. It describes using Punnett squares to determine genotypic and phenotypic ratios for crosses involving one or two traits. Chi-square tests were used to analyze if the observed ratios fit the expected Mendelian ratios. For monohybrid crosses, the observed ratios fit 1:2:1 genotypic and 3:1 phenotypic ratios. However, for some dihybrid cross samples, the observed ratios did not fit the expected 9:3:3:1 phenotypic ratio, possibly due to limitations in the experimental design and samples used.
This document presents information on complementation tests. It defines complementation tests as a method used to determine if two mutations are in the same gene or different genes. It explains that if the mutations are complementary (in different genes), the offspring will show the parental phenotypes, but if they are not complementary (in the same gene), the offspring will show a new phenotype. Three examples of using complementation test results to determine the number of genes involved are provided. The document concludes by citing a reference for more information on assigning mutations to genes using complementation tests.
Mendel performed a dihybrid cross experiment involving two traits - plant height and flower color. He crossed a tall, red-flowered homozygous plant with a dwarf, white-flowered homozygous plant. The F1 generation were all tall with red flowers, showing dominance. In the F2 generation, there were 9 tall red plants, 3 tall white plants, 3 dwarf red plants, and 1 dwarf white plant, in a 9:3:3:1 ratio. This supported Mendel's second law of independent assortment, that two traits are inherited independently. Test crosses on the F1 hybrids confirmed independent assortment of alleles between the two gene pairs.
This document describes the discovery and characterization of a common inversion polymorphism on chromosome 8p in humans. Analysis of recombination patterns in families identified apparent triple recombinations in a 12 cM region on chromosome 8p that were resolved by inverting the order of two markers. Fluorescent in situ hybridization confirmed the inversion in these families and others, estimating the inversion frequency at 21% in individuals of European ancestry. The inversion spans approximately 12 cM genetically and 2.5-5.3 Mb physically, and is flanked by clusters of olfactory receptor genes, suggesting it may be mediated by recombination between these repeats. The polymorphism could impact susceptibility to certain chromosomal abnormalities and influence gene expression near the breakpoint.
This document summarizes the discovery of a novel 162 base pair exon insertion, denoted exon 9a, in the interleukin 23 (IL-23) receptor gene. The insertion was found between exons 9 and 10 of the IL-23 receptor mRNA. Sequence analysis revealed splice donor-acceptor sites flanking the insertion, suggesting it is a legitimate cryptic exon. Although the insertion is in-frame, it contains a premature stop codon that would result in a truncated and non-functional IL-23 receptor protein. Further analysis detected the exon 9a insertion at low levels in mRNA from mitogen-stimulated immune cells, indicating it arises from rare alternative splicing of the IL-23 receptor pre-mRNA. Homologous
Poster: Effects of LEMS on P/Q type Calcium Channelsmaryvi
This study examined the effects of Lambert-Eaton myasthenic syndrome (LEMS) plasma on calcium channel expression in tottering (tg) mice, which have a mutation in the P/Q-type calcium channel subunit α1A. Western blots were used to analyze the expression of calcium channel subunits α1A, α1B, α1C, and α1E in muscle tissue from tg and wild-type mice treated with either LEMS plasma or saline solution. The results suggest that N-type and R-type calcium channels may compensate for the loss of P/Q-type channels in tg mice treated with LEMS plasma. However, the antibodies used showed low specificity, so more experiments
This document summarizes a study that identified 24 splice variants of the human IL-23 receptor (IL-23R) gene from activated human leukocytes. The variants were characterized through restriction enzyme digestion and sequencing of IL-23R cDNA clones. Most variants involved exon skipping or deletions in the extracellular region of the IL-23Ra protein, which could impact IL-23 ligand binding and signaling. The identification of these potentially functional IL-23R variants improves understanding of how the gene contributes to both normal immune function and pathological conditions like inflammatory bowel disease.
Thant Bio Symposium Poster Spring 2016Claire Thant
The study found that axonal transport defects activate the PI3K pathway in an attempt to reduce oxidative stress and neuronal cell death in neurodegenerative disease models. Expressing constitutively active PI3K decreased cell death caused by polyQ repeats and Paraquat exposure but did not affect axonal transport blockages. Dominant negative PI3K disrupted normal huntingtin motility, indicating PI3K acts downstream of transport. Motor protein mutations and disease models showed increased levels of p-GSK3β, a PI3K effector, suggesting transport defects trigger the protective PI3K response.
This study aimed to confirm previous findings linking canine chromosomes 1 and 19 to idiopathic epilepsy in Australian Shepherd dogs. PCR-RFLP tests of two SNPs were performed on 88 additional Australian Shepherds and genotypes were compared between affected and unaffected dogs via Chi-square analysis. Sequencing of the DOK6 gene did not reveal differences between one affected and unaffected dog. The results confirmed different genotype frequencies between normal and affected dogs for the SNPs, supporting the hypothesis that epilepsy-causing genes are located near the tested regions on chromosomes 1 and 19.
1) The study investigated the expression of the protein TMEM35 in different cell lines and its effects on surface expression of the nicotinic acetylcholine receptor α7 (α7 nAChR).
2) Western blot analysis showed that TMEM35 expression levels correlated with the ability of cell lines to express surface α7 nAChR after transfection.
3) Preliminary results suggest C-terminally tagged TMEM35 may not enhance α7 nAChR surface expression in the same way as untagged TMEM35, possibly due to interaction of the C-terminus with the chaperone protein RIC3.
This study investigated the role of histone H3 lysine 18 acetylation (H3K18ac) in maintaining pluripotency in embryonic stem cells. The researchers found that H3K18ac is highly enriched at stem cell genes in embryonic stem cells and levels decrease with differentiation. Expression of the E1A oncoprotein in mouse embryonic stem cells globally reduced H3K18ac and caused the cells to lose pluripotent characteristics, fail to activate stem cell enhancers, and suppress lineage-specific genes. Loss of pluripotency upon E1A expression required binding of E1A to the H3K18 acetyltransferases p300/CBP. This suggests H3K
antibody structure classification and functionsDUSHYANT KUMAR
Antibodies (immunoglobulins) are globulin proteins formed in response to antigens that specifically react with and destroy antigens. They are produced by plasma cells and found in blood serum, body fluids, and tissues. Antibodies are the immune system's way of targeting harmful foreign substances like bacteria, viruses, and more. Through experiments in 1939 and 1962, scientists discovered that antibodies are contained within the gamma globulin fraction of blood and have a basic four-chain structure. There are five classes of antibodies in human serum named for the type of heavy chain they contain: IgG, IgA, IgM, IgE, and IgD.
Molecular characterization of anaplasma platys strains from dogs in sicily, i...Josephine Huang
1) Researchers analyzed blood samples from 344 dogs in Sicily, Italy to characterize strains of Anaplasma platys.
2) They found A. platys DNA in 14 dogs (4% prevalence) through PCR and gene sequencing of the 16S rDNA, groESL, and gltA genes.
3) Sequence analysis identified at least 3 different genotypes of A. platys among the Sicilian dog samples based on variations in the gltA gene.
This document summarizes a study on the phylogeny of predatory stink bugs in the subfamily Asopinae. The analysis recovered the monophyly of Asopinae, which was supported by 16 synapomorphies. Eighteen of the 19 genera included in the analysis with multiple species were monophyletic, while the genus Podisus was paraphyletic. The pseudoclaspers found in males may have contributed to the evolutionary success of Asopinae by enhancing copulatory performance compared to other pentatomids.
Genetic polymorphism of pigs (Sus Scrofa domestica) of Ukrainian meat and Wel...Maria Drahulian
Genetic polymorphism of pigs (Sus Scrofa domestica) of Ukrainian meat and Welsh breeds according to cyto- and molecular- genetic markers /M. Drahulian, S. Kostenko, T. Dorosch // Cytopathology. Volume 29, Issue S1 Abstracts of the 41st European Congress of Cytopathology, Madrid, Spain, 10 – 13 June 2018. P - 129. DOI: 10.1111/cyt.12569
Elucidating changes in gene expression in Tryp susceptible and resistant cattle during progression of tryp infection using Affymetrix gene expression Micro arrays
Detection of ehrlichia platys dna in brown dog ticksJosephine Huang
This document reports on a study that detected Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) collected from dogs in Okinawa Island, Japan. Key findings include:
- Partial 16S rRNA gene sequences of E. platys were detected in 3 out of 32 ticks collected from 2 dogs using PCR and sequencing.
- This represents the first detection of E. platys in Japan and the first report of detecting it in ticks.
- Sequence analysis showed the 3 positive ticks were highly similar (99.7-100% identical) to known E. platys sequences, confirming the detection.
- This suggests R. sanguineus ticks
The document discusses a simulation of a population of bunnies in England, where some bunnies have a recessive allele that causes them to lack fur. Over generations, as the harsh winters cause the hairless bunnies to die off at higher rates, the frequency of the recessive allele decreases. The results demonstrate natural selection, as the recessive allele becomes less common as it confers a lower chance of survival in that environment.
This study aims to determine if differences in alleles of the APETALA1 (AP1) gene are responsible for phenotypic differences between varieties of Brassica oleracea, specifically cauliflower and Rbo. The researchers generated F1 and F2 crosses between cauliflower and Rbo, which showed segregating phenotypes. They are determining the genotypes of the AP1a and AP1c alleles in the F2 plants to test if there is a correlation between genotype and phenotype. Preliminary results found the AP1c sequence from cauliflower is identical to sequences from broccoli and kale, suggesting AP1 may not be responsible for phenotypic differences as hypothesized.
This study compared the contractile response patterns of ergot alkaloids isolated from tall fescue in bovine lateral saphenous veins. The ergoline alkaloids lysergic acid, lysergol, and ergonovine produced quicker contractile responses that began relaxing immediately, while the ergopeptine alkaloids ergovaline, ergotamine, ergocristine, ergocryptine, and ergocornine had slower, more persistent contractile responses with little to no relaxation over 120 minutes. The different classes of alkaloids produced significantly different contractile response patterns in the veins. Persistent vasoconstriction by certain ergot alkaloids may
This document discusses cattle losses due to larkspur poisoning on grazing lands in Utah. Typical cattle losses are around 5% but can reach 15%. Clinical signs of poisoning include staggering, muscle tremors, and collapse. Grazing management strategies recommend grazing early or late to avoid the "toxic window" when larkspur is most dangerous. A study evaluated cattle breeds' susceptibility and identified genetic markers associated with resistance in Angus cattle. Further research aims to validate these markers and identify other genes influencing larkspur resistance.
- One or more of the three children will have disease 1.
- All three children will have the disease.
- At least one child out of three will be phenotypically normal.
Of 150 E. coli strains cultured from cattle in Europe, 3 were resistant to colistin. One strain was found to carry the mcr-1 gene, conferring plasmid-mediated colistin resistance. This strain also showed resistance to beta-lactams, florfenicol, and fluoroquinolones. Whole genome sequencing identified resistance genes and plasmids in the 3 colistin-resistant strains. The mcr-1 gene was found on a plasmid in one strain isolated in France in 2007, demonstrating the presence of this gene in livestock in Europe.
Three farms experienced adverse reactions in neonatal pigs after ivermectin injections to control mange. Young piglets exhibited central nervous system disturbances like tremors and ataxia. Liver samples from affected piglets contained much higher levels of ivermectin than expected. The overdoses were likely due to treating very young neonates and using large syringes, making accurate dosing impossible. Producers are advised not to inject ivermectin in neonates or to dilute it and use a small syringe if treatment is necessary.
1. The document discusses blood group systems in dogs and the antibodies associated with them. The major blood group systems in dogs are DEA1-1, DEA1-2, and DEA3-8.
2. DEA1-1 and DEA1-2 antibodies are found in approximately 60% of dog populations. Dogs with these blood groups can have severe transfusion reactions if previously sensitized.
3. Other blood groups discussed include DEA1-3, DEA4, DEA7, and DAL, some of which are less immunogenic or absent in certain dogs.
Female mammals achieve dosage compensation by inactivating one of their two X chromosomes
during development, a process entirely dependent on Xist, an X-linked long noncoding
RNA (lncRNA). At the onset of X chromosome inactivation (XCI), Xist is up-regulated
and spreads along the future inactive X chromosome. Contextually, it recruits repressive
histone and DNA modifiers that transcriptionally silence the X chromosome. Xist regulation is
tightly coupled to differentiation and its expression is under the control of both pluripotency
and epigenetic factors. Recent evidence has suggested that chromatin remodelers accumulate
at the X Inactivation Center (XIC) and here we demonstrate a new role for Chd8 in Xist
regulation in differentiating ES cells, linked to its control and prevention of spurious
transcription factor interactions occurring within Xist regulatory regions. Our findings have a
broader relevance, in the context of complex, developmentally-regulated gene expression.
This document discusses multiple allelism, which refers to more than two alternative allelic forms of a gene occupying the same locus. It provides examples of multiple allelism in eye color in Drosophila, with 14 alleles producing different shades from white to red, and in human blood groups with the A, B, and O alleles. The characteristics of multiple alleles are described, including that only two alleles are present per individual. Multiple allelism in inheritance of blood groups and determining blood group combinations in offspring are also covered.
This document summarizes multiple alleles and provides examples. It discusses:
1) The human ABO blood group system which has 3 alleles (IA, IB, I0) determining 4 blood groups.
2) Inheritance of coat color in rabbits which is controlled by 4 alleles of the C gene determining 5 coat patterns.
3) Inheritance of the Rh factor in human blood which has 2 types - Rh positive and Rh negative.
- Serum carboxylesterase knockout (sCaE KO) mice lack an enzyme that provides increased protection against certain organophosphorous nerve agents in mice and rats compared to primates.
- Analysis found that sCaE KO mice are physiologically similar to wild-type mice except for the absence of the carboxylesterase enzyme in their blood.
- sCaE KO mice were found to have LD50 values for G-series nerve agents that were 20-40% of those for wild-type mice, indicating they are a more relevant model for predicting human responses compared to other small animal models.
- Injection of bioscavenger enzymes in sCaE KO mice protected them against
Lack of induced mutagenesis-Seminario Biología Molecular-Juan Restrepo RuizJuan Restrepo Ruiz
This document summarizes an experiment evaluating the effect of irradiated meat samples on E. coli and human lymphoblast cell lines. Various assays were conducted including measuring mutation frequency in E. coli, detecting antibiotic resistance plasmids, genome sequencing, and analyzing human lymphoblast mutation rates. The results found no evidence of mutagenesis from the irradiated meat samples in any of the assays. The conclusion is that food irradiation is a safe technology for food preservation and addresses misconceptions around safety.
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Lack of induced mutagenesis-Seminario Biología Molecular-Juan Restrepo Ruiz
Nemec ABT 399 Research Report final
1. Agricultural Biotechnology 399 Research Report
I. Proposal Title: Investigating Sequences of Exon 1 of the CXCL16
Candidate Gene for Equine Arteritis Virus Resistance
Among Equidae, Rhinoceridae, and Tapiridae
II. Name: Brooke Nemec
E-mail: Brooke.Nemec@uky.edu
Graduation Date: May 2015
III. Faculty Advisor: Dr. Ernest Bailey, Department of Veterinary Science
IV. Statement of Career Goals: After completing my Agricultural Biotechnology
degree at the University of Kentucky, I plan to attend veterinary school to become a
wild life or large breed veterinarian.
2. V. Abstract
Equine Viral Arteritis (EVA) is a contagious viral disease of equids caused by
Equine Arteritis Virus, an RNA virus. Prior research identified gene CXCL16 on
horse chromosome 11 (ECA11) with two alleles as having genetic influence on the
resistance of in-vitro EAV infection of T-cells. The two alleles differed by four
mutations in the first exon of CXCL16, each altering an amino acid in the first
domain of the CXCL16 protein. This project is investigates the gene sequences of
exon 1 of CXCL16 present in other families within the Perissodactyla Order
including Equidae, Rhinoceridae, and Tapiridae and identifies phylogenetic
differences in respect to CXCL16 variation.
VI.Introduction and Significance
EVA disease causes a variety of flu like symptoms making it difficult to
distinguish from other viruses. EAV is transmitted by respiratory route or venereally.
Two significant consequences of EVA are abortion in the mare and establishment of
the carrier state in the stallion making this of high interest to the prominent horse
breeding businesses in Kentucky. Mature stallions, but not intact colts, geldings or
mares, can harbor the virus in the accessory sex glands and stallions can shed the
virus in semen.
Recently, graduate student, Dr. Yun Young Go of Dr. Balasuriya’s lab,
identified a polymorphism among horses for in-vitro infection of T-cells. Go found
that horses could be divided into two groups, susceptible or resistant, based on in-
vitro susceptibility of their lymphocytes to EAV infection. Further genome wide
3. association studies showed that a gene on the ECA11 chromosome genetically
influenced this polymorphism. Subsequent work led to identification of the candidate
gene, CXCL16, in this region with two alleles. One allele was associated with
susceptibility and the other with resistance to in vitro infection of T-cells. The 2
alleles differed by 4 mutations in the first exon of CXCL16, each altering an amino
acid in the first domain of the CXCL16 protein. The trait is dominant in the respect
that susceptible horses possessed at least one copy of the variant. CXCL16 has
associated with immune response but not much else is known regarding this gene.
As mentioned, two alleles varying by four mutations have been identified in
Equus caballus (horses) of the Equidae family. Closely related to horses are the non-
horse Equids including zebras, and Asiatic Asses (Figure 1). Distantly related to
horses are Rhinoceridae (rhinos) and Tapiridae (tapirs) who share a Perissodactyla
ancestor classifying them in the same order, Perissodactyla.
Figure 1: The currently accepted phylogenetic tree of Order Perissodactyla.
4. A reference sequence for exon 1 of CXCL16 on ECA 11 has been established.
A successfully annealing primer was already created during prior research for exon 1
of CXCL16 for equines and was used again in this study. The four identified
mutations in the reference sequence and their effects on their corresponding amino
acids are shown in Figure 2. The order of amino acids found for susceptible horses
was established as phenylalanine, histidine, isoleucine, and lysine. In contrast, the
order of amino acids found in resistant horses was established as tyrosine, aspartic
acid, phenylalanine, and glutamic acid.
Figure 2: The reference sequence for CXCL16. Exon 1 is shown in bold and
mutations in question are shown in red. A’s, T’s, G’s, and C’s represent adenine,
thymine, guanine, and cytosine nucleic acids respectively.
Because there are only two alleles shown in horses for CXCL16 of the 256
combinations possible, the null hypothesis of this experiment was that the CXCL16
gene is under strong selection and other members of Perissodactyla Order have the
5. same two alleles present. Thus, the null hypothesis was that the CXCL16 gene is
under different selection pressure and other members of the Perissodactyl Order do
not have the same two alleles present.
VII. Methods
Gene sequences from distantly related species were assessed in-silico using
genome sequences available by means of public databases, specifically the Genome
10K Sample Collection Database. Available DNA samples of two horses of known
genotype, rhinos, tapirs, and non-horse equids were collected first. The two horses of
known genotype were the controls for comparison of the experiment. Horse S had the
susceptible allele and Horse R had the resistant allele. Several samples of rhino
species including Black rhinos (n=5), white rhinos (n=6), and Indian rhinos (n=3)
were used as the experimental group for the rhinoceridae family. Several samples of
non-horse equids including onagers (n=7), Hartmann’s Zebras (n=17), Grevy Zebras
(n=6), and Grant Zebras (n=7) were used as the experimental group for the non-horse
equids family. Several samples of tapir species including Baird’s Tapirs (n=5) and
Malayan Tapir (n=5) were used as the experimental group for the tapiridae family.
Exon 1 of CXCL16 was then amplified for the equus family samples using a
previously developed primer designed by graduate student, John Eberth of Dr. Ernest
Bailey’s lab, from previous ECA 11 CXCL16 studies. At first, the rhinoceridae
family was amplified using the equidae family specific primer as well but unspecific
binding occurred. To remedy this, a rhinoceridae family specific primer was then
designed by John Eberth using the reference sequence available for rhinoceridae
6. provided by the Genome 10K Sample Collection Database for use with the
rhinoceridae family in substitution of the equidae specific primer. Unfortunately there
is not currently a publically available tapiridae reference sequence for use in
developing primers so the tapiridae family samples were limited to being amplified
using the distantly related equidae family’s specific primer.
Next, samples were shipped to Eurofins for Sanger Sequencing. Then,
sequences were aligned and compared using the sequence-aligning program,
Sequencher, Nucleotide sequences within species were compared. Then, nucleotide
sequences across species and then across different families were compared to the
known Horse R and Horse S sequences. Lastly, amino acid sequences across species,
then families, were compared as well as compared to Horse R and Horse S. Amino
acid sequences shown in each species were then applied to the established
phylogenetic tree (Figure 1) to compare phylogenetic differences and similarities in
sequence conservation.
VIII. Results and Discussion
Gel electrophoresis of the samples post PCR showed successful amplification
of exon 1 of CXCL16 for the rhinos, horses, and non-horse equids using their
respective family specific primers. Tapirs were excluded from further
experimentation due to unspecific binding and possible amplification of a
contaminant when using the equidae family specific primer. Fortunately, it has been
recently found that two introns surrounding exon 1 of CXCL16 are conserved in both
rhinoceridae and equidae families. Since the sequence is conserved in these two
7. distantly related Perissodactyla, there is a chance that it is conserved in the tapiridae
family as well. This could be useful in the future development of a tapiridae family
specific primer beginning at the site of the preserved introns rather than the sites of
the original equidae and rhinoceridae primers.
Comparing the sequences among species within the rhinoceridae family, it
was found that the nucleotide sequence of exon 1 of each of black rhino was identical,
each white rhino was identical, and each Indian rhino was identical. There were
however species specific differences comparing black rhinos, to white rhinos, to
Indian rhinos confirming that the correct DNA sample was amplified and there was
no cross contamination. This was also true for other groups comparing within species
as well- all onagers were identical, Hartmann Zebra’s were identical, Grevy Zebras
were identical, and so forth but comparing across species showed they were different
due to species specific differences (Figure 3).
Figure 3: The sequence for CXCL16. Exon 1 of each species. Note that among species
the sequences found were the same but species-specific differences (circled) were found
confirming desired amplification. A’s, T’s, G’s, and C’s represent adenine, thymine,
guanine, and cytosine nucleic acids respectively.
8. When comparing nucleic acid sequences across species and families, it was
found that only Horse R had the first thymine to adenine mutation and second
cytosine to guanine mutation. All of the non-horse equids sequenced had an exon 1
sequence identical to Horse S excluding species-specific differences. Rhinos however
did have the third mutation displaying an adenine and the fourth mutation guanine
characteristic of Horse R (Figure 4).
Figure 4: The sequence for CXCL16. Exon 1 of each species. Note that across species the
four mutations (indicated by red arrows) characteristic of Horse R were not present in
entirety. Only the third and fourth mutation were shared with rhinoceridae. A’s, T’s, G’s,
and C’s represent adenine, thymine, guanine, and cytosine nucleic acids respectively.
Finally, when comparing amino acid sequences, it was found that no species
shared a similar amino acid sequence to Horse R. Horse R has an amino acid
sequence of tyrosine, aspartic acid, phenylalanine, and glutamic acid. All non-horse
equids had a sequence similar to Horse S. This sequence was phenylalanine, histidine,
isoleucine, and lysine. Despite similarities with Horse R in respect to the third and
fourth nucleotide mutations, rhinoceridae had an amino acid sequence unlike any of
the equid species. Rhinoceridae had an amino acid sequence of phenylalanine,
glutamine, phenylalanine, and glutamic acid (Figure 5).
9. Key Amino Acids
Phenylalanine F Glutamine Q
Tyrosine Y Isoleucine I
Histidine H Lysine K
Aspartic Acid D Glutamic Acid E
Figure 5: The amino acids in question produced by exon 1 for each species (red
arrows). Note that all equids share similar amino acids as Horse S and rhinoceridae
are completely different.
The amino acid sequences can then be applied to the currently established
phylogenetic tree to compare differences and similarities between species (Figure 6).
It can be interpreted that the equidae family share a common allele characteristic of
the susceptible horse but only horses, equus caballas, have the resistant allele. The
rhinoceridae family has a completely different sequence of amino acids compared to
equidae indicating that this allele is under different selective pressure. This pressure
could include different pathogens causing the genetic selection for stronger immunity
or simply random variation in the alleles from speciation. However, the exact cause
for the difference cannot be determined at this time. The tapiridae sequences were
undetermined at this time.
10. Figure 6: The amino acid sequences (indicated in red) as they appear across families.
Note that the equids share the S sequence across species within the family but outside
the family, neither the R nor S sequence appear.
In further research it will be beneficial to further investigate if the sequence is
present in this family to determine if this allele is newly evolved to the equids,
specifically the horse, or present in tapirs as well. It would also be beneficial to
continue testing more samples as only a few rhinoceridae (n=14) and tapiridae (n=10)
were available. The allele could possibly be conserved but not as prominently thus
our available samples may not correctly reflect the entire population. More samples
of non-horse equids should also be analyzed to confirm that the samples accurately
reflect the population.
11. IX. Conclusion
In conclusion, the null hypothesis stating that the CXCL16 gene is under
strong selection and other members of Perissodactyla Order have the same two alleles
present is rejected. Thus, the null hypothesis was accepted stating that the CXCL16
gene is under different selection pressure and other members of the Perissodactyl
Order do not have the same two alleles present. It was found that there was
susceptible allele conservation within equidae and the resistant allele only appeared in
equus caballus. This could indicate that the allele is newly evolved in horses.
Rhinoceridae sequences were unlike the susceptible and resistant genotypes
incidating that this family could be under different selective pressure for the CXCL16
gene.
X. Acknowledgements
Special thanks to Dr. Ernie Bailey for supervising this project and John Eberth
for designing primers and teaching necessary skills to complete research. A
special thanks also to Allison Sparling for providing test samples from associate
laboratories.
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