RESUMO
Background: Norovirus gastroenteritis is commonly an acute infection that lasts 2-3 days, but in immunocompromised patients norovirus can cause a chronic gastroenteritis lasting for years. Norovirus replicates in the gastrointestinal tract, but the pathway of viral clearance is not yet known. Promising results of enterally administered immunoglobulin in the treatment of chronic norovirus gastroenteritis in immunocompromised patients have previously been published. Case presentation: We report two individuals with common variable immunodeficiency and chronic debilitating norovirus gastroenteritis. Both patients were treated with enterally administered immunoglobulin via a duodenal feeding tube as other treatment modalities have been unsuccessful. The patients did not experience any immediate or long-term benefit of enterally administered immunoglobulin. Conclusion: Despite previous case reports of successful treatment of chronic norovirus infection among immunocompromised patients with enterally administered immunoglobulin, these two patients experienced no benefit of the treatment. This demonstrates the need for further research in treatment of chronic norovirus infection in immunocompromised patients.
RESUMO
Small-dose, multiple injections of GnRH given to seasonally anoestrous ewes induce final stages of the preovulatory follicle development, but result in an high incidence of defective CL unless animals are primed with progesterone, which completely eliminates luteal dysfunction. Progesterone priming upregulates luteal vascularization; however, its effect on follicular angiogenesis is poorly understood. This study tested the hypothesis that progesterone priming of seasonally anoestrous ewes treated with dose multiple injections of GnRH eliminates defective luteal function by altering the expression of vascular endothelial growth factor (VEGF), VEGF receptor-2, angiopoietin (ANG)-1, ANG-2, and TIE-2 during early and late preovulatory follicle development. Ten seasonally anoestrous ewes were given 20 mg of progesterone im 3 days before the start of GnRH treatment; 10 other animals served as controls. Intravenous injections of 500 ng GnRH were given to all animals every 2 hours for 28 hours, followed at 30 hours with a 300-µg GnRH bolus injection to synchronize the preovulatory LH surge. Ovaries were collected at 24 and 46 hours after the start of GnRH treatment. Small (2-2.5 mm) and large (>2.5 mm) follicles were analyzed for protein and mRNA expression of the angiogenic factors using immunohistochemistry and in situ hybridization assays. Progesterone priming did not have an influence on angiogenic factor levels in small follicles. However, progesterone-primed animals showed significantly (P ≤ 0.05) higher levels of VEGF, VEGFR-2, ANG-1, and ANG-2 in large follicles compared with nonprimed ones. These data suggest that progesterone priming alters the expression of angiogenic factors in large preovulatory follicles, ensuring adequate luteal development and function.
Assuntos
Indutores da Angiogênese/metabolismo , Folículo Ovariano/efeitos dos fármacos , Progesterona/farmacologia , Ovinos/fisiologia , Angiopoietina-1/metabolismo , Angiopoietina-2/metabolismo , Animais , Feminino , Imuno-Histoquímica , Folículo Ovariano/metabolismo , Receptor TIE-2/metabolismo , Estações do Ano , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Gonadotrophin releasing hormone (GnRH)-induced ovulation in seasonally anestrous ewes is associated with a high incidence of defective corpora lutea (CL), which can be completely eliminated by priming ewes with progesterone before GnRH treatment, but the physiological basis of this has remained elusive. This study tested the hypothesis that progesterone priming eliminates defective luteal function by altering the expression of Vascular Endothelial Growth Factor (VEGF), its receptor VEGFR-2, and angiopoietin (ANG)-1, ANG-2 and their receptor TIE-2 in the early CL. Fifteen seasonally anestrous ewes were treated by i.m. injection with 20 mg of progesterone 3 days before the start of GnRH treatment, while another 15 animals served as controls. Intravenous injections of 500 ng GnRH were given to all the ewes every 2 h for 28 h, followed by a 300 µg GnRH bolus injection to synchronize the preovulatory luteinizing hormone (LH) surge. Corpora lutea were collected 1, 2 and 4 days after ovulation and analyzed for protein and mRNA expression of VEGF, VEGFR-2, ANG-1, ANG-2 and Tie-2 using Western Immunoblotting and in situ hybridization. VEGF, VEGFR-2 and ANG-1 expression was significantly higher (P ≤ 0.05) in the CL of progesterone-primed animals compared to non-primed ones. However, no differences were observed in the ANG-2 or Tie-2 expression levels between the two treatment groups. These data suggest that progesterone priming of the preovulatory follicle alters the expression of some angiogenic growth factors in the early CL, leading to greater vascular stability and thereby normal luteal function.
Assuntos
Angiopoietina-1/metabolismo , Corpo Lúteo/efeitos dos fármacos , Ciclo Estral/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Progesterona/farmacologia , Ovinos/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Angiopoietina-1/genética , Animais , Corpo Lúteo/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hormônio Luteinizante/sangue , Ovulação/metabolismo , Indução da Ovulação/métodos , Indução da Ovulação/veterinária , Estações do Ano , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
Bacteriophage lambda has been in use as a cloning vector for over 25 years, and has been used extensively as an expression vector. The efficiency of packaging and infection, and the simplicity of plaque screening are advantages of lambda as a cloning vector. A number of ingenious modifications help overcome the disadvantages associated with its mode of growth and its size. Some lambda vectors have been designed to be readily converted into plasmids or phagemids, and there are a variety of promoters and fusions that can be used to drive expression of foreign genes. Screening lambda libraries with antibodies or ligands is a powerful way of identifying novel genes.
Assuntos
Bacteriófago lambda/genética , Técnicas Genéticas , Vetores Genéticos , Clonagem Molecular , DNA/metabolismo , DNA Complementar/metabolismo , Biblioteca Gênica , Regiões Promotoras Genéticas , Proteínas Recombinantes de Fusão/metabolismoRESUMO
I have used an exercise involving domestic cats in the General Genetics course at the University of Nebraska-Lincoln for the past 5 years. Using a coherent set of traits in an organism familiar to the students makes it easy to illustrate principles of transmission and population genetics. The one-semester course consists primarily of sophomores and juniors who have either taken a one-semester introductory biology course, a one-semester cell biology course, or have a strong high school biology background. The students are given a handout and asked to determine the genotype at seven unlinked loci of at least one cat. To fill out the form, the students have to grasp such concepts as dominance, incomplete dominance, temperature-sensitive mutations, epistatic interactions, sex linkage, and variable expressivity. Completing the form reinforces these concepts as they observe the cat's phenotype and fill in the genotype. I then analyze the collected data and use it in my lectures on population genetics to illustrate the Hardy-Weinberg equilibrium, calculate allele frequencies, and use statistics. This allows the students to look at population genetics in a very positive light and provides concrete examples of some often misunderstood principles.
Assuntos
Gatos/genética , Epistasia Genética , Genes Dominantes , Genes Recessivos , Genética/educação , Ensino/métodos , Alelos , Animais , Feminino , Frequência do Gene , Genética Populacional , Genótipo , Cor de Cabelo/genética , Masculino , Fenótipo , Fatores Sexuais , Estados UnidosRESUMO
We have cloned and sequenced a cDNA from Drosophila melanogaster that encodes a protein homologous to the peritrophins, a family of chitin-binding proteins from the peritrophic matrix of insects. Unexpectedly, the gene, Gasp, is expressed in the embryonic tracheae. We suggest that this family of proteins may be present in other tissues than the peritrophic matrix, particularly where nutrient or gas exchange are important, and/or where invasion by parasites or viruses is possible. We have also mapped two similar genes that had been sequenced by the Berkeley Drosophila Genome Project, and find that these three very similar genes are not clustered, but are located on three different chromosomes.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster/genética , Proteínas de Insetos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar , Drosophila melanogaster/embriologia , Expressão Gênica , Dados de Sequência Molecular , Análise de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Traqueia/embriologiaAssuntos
Bacteriófago lambda/genética , Clonagem Molecular/métodos , Vetores Genéticos/genética , Proteínas Recombinantes de Fusão/biossíntese , Bacteriófago T7/genética , Células Eucarióticas/fisiologia , Expressão Gênica , Biblioteca Gênica , Óperon Lac , Regiões Promotoras Genéticas , Seleção GenéticaRESUMO
AIMS: The aim of this study was to find an algorithm of better fit for early eye growth than the linear regression usually advanced. METHODS: The analysis is based on previously published around term data, the main material being axial ultrasound measurements in preterm (n = 101) and full term infants (n = 25). The postconceptional age of the infants ranged between 36 and 54 weeks. Previously published Danish data from eyes of aborted fetuses were also used, as were averaged values from the literature regarding eye size at age 1 year (20 mm), 3 years (22 mm), and a presumed 13 year endpoint of 23 mm. RESULTS: A second order exponential function fitted with the basic data within a standard deviation of 2%. CONCLUSIONS: A simple symbolic expression and tabulated values for eye growth in infancy and childhood were derived. This is clearly of practical value, for example, when following the development of eyes treated for congenital glaucoma or assessing other developmental anomalies and early eye diseases.
Assuntos
Olho/crescimento & desenvolvimento , Algoritmos , Pré-Escolar , Olho/diagnóstico por imagem , Feminino , Humanos , Lactente , Recém-Nascido , Gravidez , UltrassonografiaRESUMO
In the genome of Drosophila melanogaster there is a single locus, Triplo-lethal (Tpl), that causes lethality when present in either one or three copies in an otherwise diploid animal. Previous attempts to mutagenize Tpl produced alleles that were viable over a chromosome bearing a duplication of Tpl, but were not lethal in combination with a wild-type chromosome, as deficiencies for Tpl are. These mutations were interpreted as hypomorphic alleles of Tpl. In this work, we show that these alleles are not mutations at Tpl; rather, they are dominant mutations in a tightly linked, but cytologically distant, locus that we have named Suppressor-of-Tpl (Sul(Tpl)). Su(Tpl) mutations suppress the lethality associated with three copies of the Triplo-lethal locus and are recessive lethal. We have mapped Su(Tpl) to the approximate map position 3-46.5, within the cytological region 76B-76D.
Assuntos
Drosophila melanogaster/genética , Genes de Insetos , Genes Letais , Genes Supressores , Animais , Sequência de Bases , Mapeamento Cromossômico , Elementos de DNA Transponíveis , Deleção de Genes , Regulação da Expressão Gênica no Desenvolvimento , Genes Recessivos , Teste de Complementação Genética , Ligação Genética , Dados de Sequência Molecular , Mutagênese , Recombinação GenéticaRESUMO
An organism that seems to be identical to Orskov's 'Sarcina mirabilis' [Orskov, J. (1930) Acta Pathol Microbiol Scand Suppl III, 519-541] has been rediscovered in specimens from the upper respiratory tract of humans. Six strains were studied, and the results, which conformed to Orskov's description of S. mirabilis, were as follows. Rough to smooth colonies grow on many plated media and show extremely polymorphic cell morphology with round cells with diameters from 1 to > 10 microns. The smallest cells were often motile with circular movements. Strains were Gram-negative, facultatively anaerobic, oxidase and urease positive, and weakly catalase positive. Nitrate and nitrite were reduced, and glucose, fructose, sucrose and mannitol were fermented. Polysaccharide was produced on sucrose agar. Electron microscopy showed coccoid cells with a bundle of three to nine flagella, a Gram-negative cell-wall morphology, and aggregates of irregular cells held together by a common surface layer. The mean mol% (G+C) of the organisms was 65.0. 16S-ribosomal RNA sequencing revealed that the organism belongs to the beta subgroup of Proteobacteria, separate from all other described genera, but most closely related to Burkholderia. The name Lautropia mirabilis is proposed for this organism.
Assuntos
Cocos Anaeróbios Gram-Negativos/classificação , Cocos Anaeróbios Gram-Negativos/isolamento & purificação , Boca/microbiologia , Técnicas de Tipagem Bacteriana , Divisão Celular , Movimento Celular , Cocos Anaeróbios Gram-Negativos/genética , Cocos Anaeróbios Gram-Negativos/ultraestrutura , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
One of the most extreme examples of gene dosage sensitivity is the Triplo-lethal locus (Tpl) on the third chromosome of Drosophila melanogaster, which is lethal when present in either one or three copies. Increased dosage of an unlinked locus, Isis, suppresses the triplo-lethal phenotype of Tpl, but not the haplo-lethal phenotype. We have mapped Isis to the X chromosome region 7E3-8A5, and shown that the suppression is a gene dosage effect. Altered dosage of Isis in the presence of two copies of Tpl has no obvious effects. By examining the interactions between Isis dosage and Tpl we suggest that Isis does not directly repress Tpl expression, but acts downstream on the triplo-lethal phenotype of Tpl.
Assuntos
Drosophila melanogaster/genética , Animais , Mapeamento Cromossômico , Feminino , Genes Letais , Ligação Genética , Masculino , Família Multigênica , Mutação , Fenótipo , Supressão Genética , Translocação Genética , Trissomia , Cromossomo X , Cromossomo YRESUMO
A locus associated with a severe Minute effect has been mapped at 7C on the X chromosome of Drosophila melanogaster. Previous work has suggested that this Minute encodes ribosomal proteins S14A and S14B. We have made a chromosomal deficiency that removes the S14 ribosomal protein genes, yet does not display the Minute phenotype. These data suggest that the S14 genes do not actually correspond to the Minute locus.
Assuntos
Drosophila melanogaster/genética , Proteínas Ribossômicas/genética , Animais , Bandeamento Cromossômico , Mapeamento Cromossômico , Cruzamentos Genéticos , DNA/genética , Feminino , Cariotipagem , Masculino , Mutagênese , Hibridização de Ácido Nucleico , Fenótipo , Cromossomo XRESUMO
Vision is a highly complex process common to many vertebrate and invertebrate species. The visual system of Drosophila represents a valuable model in which to study this process since its chromosomal makeup has been well mapped and many mutants containing specific photoreceptor defects have been described. S-antigen is a well-characterized photoreceptor cell protein which plays a fundamental role in the visual process. In this study, we used a panel of four monoclonal antibodies (MAbs) specific for amino, mid and carboxy terminal amino acid sequences in S-antigen in order to examine S-antigen immunoreactivity in the Drosophila visual system. Two MAbs, MAb5C6.47 and MAbC10-C10, localized S-antigen to the retinules and ocelli. In contrast, MAbA2-G5 and MAbA9-C6 did not stain. The presence and strong antigenic similarity of an S-antigen homolog in Drosophila suggests that in both vertebrates and invertebrates S-antigen plays a major role in the regulation of the phototransduction of vision.
Assuntos
Antígenos/análise , Drosophila melanogaster/imunologia , Proteínas do Olho/análise , Sequência de Aminoácidos , Animais , Arrestina , Autoantígenos/análise , Western Blotting , Imuno-HistoquímicaRESUMO
The Triplo-lethal (Tpl) locus of Drosophila is the only known locus which is lethal when present in three copies rather than the normal two. After recovering a hybrid-dysgenesis-induced mutation of Tpl we used a rapid combination of transposon tagging, chromosome microdissection and PCR to clone the P element that had transposed into the Tpl region. That P element is located within the gene for a new and unique member of the RNA helicase family. This new helicase differs from all others known by having glycine-rich repeats at both the amino and carboxyl termini. Curiously, genetic analysis shows that the P element inserted into this gene is not responsible for the Tpl mutant phenotype. We present possible explanations for these findings.
Assuntos
Elementos de DNA Transponíveis , Drosophila/genética , Ligação Genética , RNA Nucleotidiltransferases/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA/genética , Drosophila/enzimologia , Éxons , Genes , Íntrons , Dados de Sequência Molecular , Mutação , Hibridização de Ácido Nucleico , Fenótipo , Reação em Cadeia da Polimerase , Mapeamento por Restrição , Transcrição GênicaRESUMO
The Triplo-lethal locus (Tpl) is unique in its dosage sensitivity; no other locus in Drosophila has been identified that is lethal when present in three doses. Tpl is also haplo-lethal, and its function is still a mystery. Previous workers have found it nearly impossible to mutationally inactive Tpl other than by completely deleting the chromosomal region in which Tpl resides (83DE). We have utilized P-M hybrid dysgenesis in an effort to obtain new mutations of Tpl. We recovered 19 new duplications of Tpl, 15 hypomorphic mutations of Tpl (a previously rare class of mutation), and no null mutations. Surprisingly, 14 of the 15 hypomorphic alleles have no detectable P element sequences at the locus. The difficulty in recovering null mutations in Tpl suggests that it may be a complex locus, perhaps consisting of several genes with redundant functions. The relative ease with which we recovered hypomorphic alleles is in sharp contrast to previous attempts by others to mutagenize Tpl. A higher mutation rate with hybrid dysgenesis than with radiation or chemicals also suggests a peculiar genetic organization for the locus.
Assuntos
Drosophila melanogaster/genética , Genes Letais , Mutação , Alelos , Animais , Bandeamento Cromossômico , Cruzamentos Genéticos , Elementos de DNA Transponíveis , Feminino , MasculinoRESUMO
In the genome of Drosophila melanogaster there is only one locus, Tpl, that is triplo-lethal; it is also haplo-lethal. Previous work has identified 3 hypomorphic alleles of Tpl which rescue animals carrying a duplication of Tpl, but which are not dominant lethals as null mutations or deficiencies would be. We have found that all three hypomorphic alleles act as site-specific hotspots for recombination when heterozygous with a wild-type homolog. Recombination between the flanking markers ri and Ki is increased 6.5-10.5-fold in the presence of Tpl hypomorphic alleles. The increased recombination was found to occur between Tpl and Ki, while recombination in other adjacent regions is unchanged. The use of isogenic Tpl+ controls, and the use of flanking intervals in the mutant chromosomes allows us to rule out the interchromosomal effect as a cause. We have also observed premeiotic recombination occurring at the Tpl hypomorphic alleles in male heterozygotes. We hypothesize that transposons are responsible for both the hypomorphic phenotype and the high frequency of recombination.
Assuntos
Drosophila melanogaster/genética , Genes Letais , Recombinação Genética , Alelos , Animais , Elementos de DNA Transponíveis , Feminino , Masculino , FenótipoRESUMO
Using a heterologous rat cDNA probe, we have identified a 14.7 kbp Drosophila melanogaster genomic clone containing the X-linked gene Pgd+, which encodes the enzyme 6-phosphogluconate dehydrogenase (6PGD). We used in situ hybridization to larval polytene chromosomes, a somatic transient expression assay for enzyme activity, and the rescue of the lethal Pgd- phenotype by germline transformation to verify the identity of the gene. A 7.4 kbp fragment including the gene and approximately 1.2 kbp of upstream and 1.8 kbp of downstream sequences was relocated to autosomal ectopic sites by germline transformation; this transduced gene exhibits levels of enhanced activity in males comparable to those of the indigenous gene at its normal X chromosome locus. We conclude that the sequences responsible for dosage compensation of Pgd+ are included in this fragment.