RESUMO
PURPOSE: Progressive rod-cone degeneration (prcd) is an autosomal recessive retinal degeneration of dogs characterized by abnormalities in lipid metabolism. It has recently been mapped to the centromeric region of canine chromosome 9, homologous to human 17q, which contains the apolipoprotein H (apoH, protein; APOH, gene) gene involved in lipid metabolism and regulation of triglycerides. The present study was undertaken to evaluate APOH as a positional candidate for prcd. METHODS: Expression of APOH in the retina was examined by reverse transcription-polymerase chain reaction (RT-PCR) and by immunocytochemistry in normal and prcd-affected dogs. The level of apoH in the plasma was determined by western blot analysis. Intragenic polymorphic markers were identified and typed in the prcd pedigree. Canine-rodent hybrid cell lines were analyzed to detect canine APOH. RESULTS: ApoH has been localized to the photoreceptor outer segment layer by immunocytochemistry. Its expression in the retina of normal and prcd-affected dogs was confirmed by RT-PCR. The levels of antihuman apoH cross-reacting material in plasma were similar in all dogs, regardless of disease status. Finally, linkage analysis of the APOH gene with the disease locus in the prcd pedigree detected 3 recombinants among 70 informative offsprings (lod score 15.09 at 0 = 4.3 centimorgan [cM]). CONCLUSIONS: APOH is expressed in the retina and tightly linked to the prcd locus. However, despite its potential role in phenotypes of abnormal lipid metabolism associated with prcd, the gene has been excluded as a primary candidate for prcd by linkage analysis.
Assuntos
Doenças do Cão/genética , Glicoproteínas/genética , Degeneração Retiniana/veterinária , Animais , Sequência de Bases/genética , Western Blotting , Linhagem Celular , Mapeamento Cromossômico , Doenças do Cão/metabolismo , Cães , Expressão Gênica/fisiologia , Ligação Genética/genética , Glicoproteínas/metabolismo , Células Híbridas , Imuno-Histoquímica , Dados de Sequência Molecular , Linhagem , Polimorfismo Genético/genética , Retina/metabolismo , Degeneração Retiniana/genética , Degeneração Retiniana/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Roedores , beta 2-Glicoproteína IRESUMO
Photoreceptor dysplasia (pd) is one of a group of at least six distinct autosomal and one X-linked retinal disorders identified in dogs which are collectively known as progressive retinal atrophy (PRA). It is an early onset retinal disease identified in miniature schnauzer dogs, and pedigree analysis and breeding studies have established autosomal recessive inheritance of the disease. Using a gene-based approach, a number of retina-expressed genes, including some members of the phototransduction pathway, have been causally implicated in retinal diseases of humans and other animals. Here we examined seven such potential candidate genes (opsin, RDS/peripherin, ROM1, rod cGMP-gated cation channel alpha-subunit, and three subunits of transducin) for their causal association with the pd locus by testing segregation of intragenic markers with the disease locus, or, in the absence of informative polymorphisms, sequencing of the coding regions of the genes. Based on these results, we have conclusively excluded four photoreceptor-specific genes as candidates for pd by linkage analysis. For three other photoreceptor-specific genes, we did not find any mutation in the coding sequences of the genes and have excluded them provisionally. Formal exclusion would require investigation of the levels of expression of the candidate genes in pd-affected dogs relative to age-matched controls. At present we are building suitable informative pedigrees for the disease locus with a sufficient number of meiosis to be useful for genomewide screening. This should identify markers linked to the disease locus and eventually permit progress toward the identification of the photoreceptor dysplasia gene and the disease-causing mutation.
Assuntos
Doenças do Cão/genética , Glicoproteínas de Membrana , Células Fotorreceptoras de Vertebrados/patologia , Degeneração Retiniana/veterinária , Animais , GMP Cíclico/genética , Canais de Cátion Regulados por Nucleotídeos Cíclicos , Doenças do Cão/patologia , Cães , Proteínas do Olho/genética , Feminino , Marcadores Genéticos , Genótipo , Proteínas de Filamentos Intermediários/genética , Canais Iônicos/genética , Masculino , Proteínas de Membrana/genética , Proteínas do Tecido Nervoso/genética , Linhagem , Periferinas , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Degeneração Retiniana/patologia , Rodopsina/genética , Segmento Externo da Célula Bastonete/fisiologia , Transducina/genéticaRESUMO
PURPOSE: Progressive retinal atrophy (PRA) represents a heterogeneous group of retinal dystrophies, distinct forms of which occur in different canine breeds. The present study was undertaken to evaluate the gene for the alpha-1 subunit of the rod specific G-protein transducin (GNAT1), a member of the phototransduction pathway, as a candidate for progressive rod cone degeneration (pred) in poodles, early retinal degeneration (erd) in elkhounds, and rod cone dysplasia 2 (rcd2) in collies. METHODS: Oligonucleotide primers were designed from the consensus region of known cDNA sequences for GNAT1 from other species. Canine GNAT1 cDNA was cloned and sequenced after reverse transcription (RT) and polymerase chain reaction (PCR) of total retinal RNA, and PCR amplification of specific sequences from a canine retinal cDNA library. Large, intron containing fragments of the canine transducin alpha-1 subunit gene were amplified from genomic DNA of individuals in PRA informative pedigrees, using canine-specific primers. PCR products were digested with Nci I, to enable typing of individuals in the PRA affected pedigrees for a previously identified GNAT1 restriction fragment length polymorphism (RFLP). RESULTS: The sequence of canine GNAT1 cDNA is reported (GenBank accession no. U65376). Over the coding region, the canine GNAT1 cDNA sequence presented here shares 92-95% identity with human, bovine and murine sequences. The canine cDNA encodes a polypeptide of 350 amino acids; its theoretical translation is 98-99% identical with the corresponding GNAT1 sequence from each of the other 3 species and it has no unique amino acids. In rcd2 and erd pedigrees informative for both the disease locus and the GNAT1 Nci I RFLP, a minimum of 3 and 2 recombinants were identified, respectively. Similarly, in a prcd pedigree, 3 of 7 progeny informative for both prcd and this RFLP were obligate recombinants. CONCLUSIONS: The canine GNAT1 gene has been excluded as a candidate for prcd, erd and rcd2. Sequence information of canine GNAT1 gene will enable testing this locus as a candidate in other canine hereditary retinal degenerations.
Assuntos
DNA Complementar/genética , Retina/patologia , Degeneração Retiniana/genética , Células Fotorreceptoras Retinianas Bastonetes/química , Transducina/genética , Sequência de Aminoácidos , Animais , Atrofia/genética , Sequência de Bases , Bovinos , Clonagem Molecular , Primers do DNA/química , Cães , Eletroforese em Gel de Poliacrilamida , Feminino , Ligação Genética , Genótipo , Humanos , Masculino , Camundongos , Dados de Sequência Molecular , Linhagem , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Degeneração Retiniana/patologia , Homologia de Sequência de Aminoácidos , Transducina/isolamento & purificaçãoRESUMO
PURPOSE: To develop an improved diagnostic test for rod-cone dysplasia type 1 (rcd1). The rcd1 phenotype is an early onset, autosomal recessive disease caused by a mutation in the canine rod cyclic GMP phosphodiesterase beta-subunit (PDE6B) gene. A G to A transition in codon 807 at nucleotide position 2420 results in a stop codon. This is the only disease causing mutation detected so far in the canine PDE6B gene. METHODS: Allele specific primers were designed in which the 3 end had the nucleotide corresponding to either the wild type or the mutant rcd1 allele. PCR was done using the allele specific primers in combination with a common primer complementary to the opposite strand to distinguish between the wild type and the rcd1 alleles. RESULTS: The wild type and rcd1 alleles were identified successfully in two independent ASPCRs done with two different sets of allele specific primers. Further, both alleles could be amplified in a single tube and distinguished based on the size difference of the PCR products using one allele specific primer of altered length by the addition of a 9 nucleotide long linker. CONCLUSIONS: We have developed an improved diagnostic test for the disease based on ASPCR such that the presence or absence of different size amplified fragments provides direct determination of the genotype. In contrast to previously reported diagnostic tests, this method is more efficient because it eliminates the need for any further manipulation of the PCR product.
Assuntos
Doenças do Cão/diagnóstico , Doenças do Cão/genética , Diester Fosfórico Hidrolases , Reação em Cadeia da Polimerase/métodos , Degeneração Retiniana/veterinária , 3',5'-GMP Cíclico Fosfodiesterases/genética , Alelos , Animais , Sequência de Bases , Nucleotídeo Cíclico Fosfodiesterase do Tipo 6 , Primers do DNA , Doenças do Cão/enzimologia , Cães , Estudos de Avaliação como Assunto , Genes Recessivos , Mutação , Fenótipo , Degeneração Retiniana/diagnóstico , Degeneração Retiniana/genéticaRESUMO
Rod-cone dysplasia type 1 (rcd1) is one of several canine photoreceptor degenerations, collectively termed progressive retinal atrophy (PRA), that afflict different breeds of dogs. The rcd1 phenotype is an early onset autosomal recessive disease caused by a nonsense amber mutation, at codon 807, in the canine gene for the beta-subunit of rod cyclic GMP phosphodiesterase (canine PDEB). The mutation involves a G to A transition at nucleotide position 2420, which presumably would cause premature termination of the canine PDEB protein by 49 amino acid residues. In both a small pedigree study of Irish setters from the United Kingdom and in larger canine pedigree studies in the United States, this gene defect has been found to be the only mutation causing rcd1. Here we report development of a diagnostic test which unequivocally distinguishes the three genotypes at the rcd1 locus: rcd1/rcd1 (homozygous mutant, affected); rcd1/+ (heterozygous, carrier); and +/+ (homozygous normal, wildtype).
Assuntos
Doenças do Cão/diagnóstico , Biologia Molecular/métodos , Células Fotorreceptoras/anormalidades , Degeneração Retiniana/genética , Degeneração Retiniana/veterinária , 3',5'-GMP Cíclico Fosfodiesterases/genética , Animais , Sequência de Bases , Análise Mutacional de DNA , Primers do DNA/química , Doenças do Cão/genética , Cães , Genótipo , Dados de Sequência Molecular , Mutação Puntual/genética , Reação em Cadeia da Polimerase/veterinária , Degeneração Retiniana/diagnósticoRESUMO
PURPOSE: To determine if a previously reported nonsense mutation (G to A transition at nucleotide position 2420) in the canine rod cyclic GMP (cGMP) phosphodiesterase beta (PDEB) subunit gene cosegregates with the rod-cone dysplasia 1 disease allele (rcd1) in the rcd1-dog reference colony; to establish the prevalence of this mutation among rcd1-affected Irish setters in the United States; and to screen for this mutation in other forms of canine hereditary progressive retinal atrophy (PRA). METHODS: Exon 21 of canine PDEB, previously reported to contain a nonsense mutation in rcd1-affected dogs, was amplified by polymerase chain reaction from genomic DNA isolated from peripheral blood samples. The mutation was detected in amplified DNA by restriction enzyme digestion and double-stranded conformational polymorphism. Linkage between rcd1 and the PDEB mutation was tested using the computer program LIPED: RESULTS: Three different rcd1-informative canine pedigrees were tested for the PDEB nonsense mutation. The first was a multigenerational pedigree representing the rcd1 reference colony. The other two pedigrees represented purebred Irish setter breeding lines in which rcd1 was known to be segregating. In all three pedigrees, the same point mutation was present and segregated with no discordance with the rcd1 allele. Linkage analysis established a maximum logarithm of odds (LOD) score of 12.05 at a linkage distance (theta) of 0.0. In a representative sampling of Irish setters in the United States diagnosed clinically as affected with typical rcd1 phenotype, all dogs were demonstrated to have the same (codon 807) PDEB mutation. Three of four Irish setters affected with atypical, relatively slower disease also had this mutation, but one dog did not. This point mutation in the canine PDEB gene was absent in other forms of canine hereditary retinal degeneration. CONCLUSIONS: In three informative pedigrees, the codon 807 mutation in canine PDEB cosegregates with the rcd1 disease allele with zero discordance. A linkage distance (theta) of zero, with an LOD score of 12.05, indicates identity of this mutation and rcd1. This appears to be the only mutation causing rcd1 in the United States. In all other forms of canine hereditary retinal degeneration tested (cd, erd, prcd, rcd2, X-linked PRA, and in one Iris degeneration tested (cd, erd, prcd, rcd2, X-linked PRA, and in one Irish setter with late onset PRA), this PDEB point mutation was absent.
Assuntos
3',5'-GMP Cíclico Fosfodiesterases/genética , Códon/genética , Doenças do Cão/genética , Mutação , Células Fotorreceptoras/patologia , Degeneração Retiniana/veterinária , Células Fotorreceptoras Retinianas Bastonetes/enzimologia , Animais , Atrofia , Sequência de Bases , DNA/análise , Doenças do Cão/patologia , Cães , Feminino , Ligação Genética/genética , Escore Lod , Masculino , Dados de Sequência Molecular , Linhagem , Reação em Cadeia da Polimerase , Prevalência , Degeneração Retiniana/genética , Degeneração Retiniana/patologiaRESUMO
Necrotizing funisitis is an umbilical cord lesion characterized by perivascular bands of necrotic Wharton jelly containing inflammatory cells in various stages of degeneration. Sixty cases were reviewed histologically. Clinical information was available in 45. Forty-five age-matched infants with acute (nonspecific) funisitis only were used as controls. Infants with necrotizing funisitis had more stillbirths, birth weights below the tenth percentile (small for gestational age [SGA]), infectious complications, and necrotizing enterocolitis. No consistent infectious agents or predisposing maternal factors were found. Cord neovascularization correlated with SGA infants. Necrotizing funisitis occurred in 0.1% of deliveries greater than 20 weeks' gestation. The perivascular bands, likened to the pattern of an Ouchterlony diffusion plate, suggest the presence of a diffusible toxin in the amniotic fluid. The stillbirths and SGA infants may represent the toxin's effect on the fetus. The lack of perivascular necrotic bands around vessels on the placental surface suggests neutralization or more effective clearing of the agent in this region, for reasons as yet undetermined. The factors underlying the cord lesion may contribute to superimposed acute nonspecific vasculitis and chorioamnionitis.
Assuntos
Doenças Fetais/patologia , Cordão Umbilical/patologia , Doença Aguda , Peso ao Nascer , Feminino , Doenças Fetais/epidemiologia , Seguimentos , Humanos , Recém-Nascido , Doenças do Recém-Nascido/epidemiologia , Doenças do Recém-Nascido/etiologia , Doenças do Recém-Nascido/mortalidade , Inflamação/epidemiologia , Inflamação/patologia , Necrose , Doenças Placentárias , Gravidez , Complicações na Gravidez , PrevalênciaRESUMO
In three fetuses, congenital intestinal atresia was associated with linear ulcerations of the umbilical cord. In two cases, hemorrhage was seen from the cord ulcer. Both fetuses required emergency cesarean section for fetal distress and were born anemic. The third fetus was mildly hydropic, attributed to hemorrhage, and was stillborn. The mechanism of the association could not be determined. These cases suggest a risk of prenatal umbilical cord hemorrhage in infants with intestinal atresia.
Assuntos
Obstrução Duodenal/congênito , Doenças Fetais/patologia , Atresia Intestinal/patologia , Jejuno/anormalidades , Cordão Umbilical , Adulto , Obstrução Duodenal/patologia , Feminino , Hemorragia/congênito , Hemorragia/etiologia , Hemorragia/patologia , Humanos , Recém-Nascido , Jejuno/patologia , Masculino , Úlcera/complicações , Úlcera/congênito , Úlcera/patologia , Cordão Umbilical/patologiaRESUMO
A brother and a sister presented with a malformation syndrome consisting of facial anomalies, distal arthrogryposis with camptodactyly of fingers and "hammer toes," severe mental retardation, and hypopituitarism. The girl is now 6 1/2 years old and exhibits severe mental retardation. She has abnormal secretion of growth hormone and responded to growth hormone therapy. Her brother was born with the same facial manifestations, distal contractures, and hypopituitarism. He died unexpectedly at age 3 months of no apparent cause. The occurrence of the syndrome in 2 sibs of different sex suggests autosomal recessive inheritance.
Assuntos
Anormalidades Múltiplas/genética , Artrogripose/genética , Face/anormalidades , Hipopituitarismo/genética , Deficiência Intelectual/genética , Criança , Feminino , Genes Recessivos , Transtornos do Crescimento/genética , Humanos , Lactente , Masculino , SíndromeRESUMO
We report on 2 patients who were apparently normal at birth but later developed characteristics of Wiedemann-Beckwith syndrome (WBS). Both had hypoglycemia neonatally and gradually developed coarse facial changes, umbilical hernia, and macroglossia. Renal sonography done after the macroglossia developed showed large kidneys in both. The placentas were carefully examined in both cases but findings described as typical of WBS were only found in one. The clinical evolution of these infants suggests that some WBS manifestations may have their onset postnatally in some cases. We postulate that the cellular hyperplasia and hypertrophy characteristic of WBS may be caused by persistent rests of embryonal cells that secrete paracrine and/or endocrine growth factors.
Assuntos
Síndrome de Beckwith-Wiedemann/diagnóstico , Humanos , Hipoglicemia , Lactente , Recém-Nascido , Rim/diagnóstico por imagem , Masculino , RadiografiaRESUMO
Selective intervention in multiple pregnancy is being used to enhance the chances of survival of at least one conceptus when the risks for the combined conceptuses and mother are considered too great. These procedures have been applied to induced polyembryonic conceptions (selective continuance) and discordant dichorionic twins (selective birth). We report attempts at selective intervention in three monochorionic twin gestations affected by twin-to-twin transfusion syndrome. In all three cases, both fetuses seemed doomed and the mother was in significant distress. The selected survivor in the first case is doing well; both twins were stillborn in the second case; in the third case, the selected survivor died as a neonate but the other twin survived and is doing well. We suggest possible explanations for the clinical outcome of each case based on detailed pathologic examination of the delivered placentas and autopsy examination of the nonsurviving twins. The shared chorionic circulation is the source of both the clinical disorder and the potential complications of any attempt to alleviate the disorder. This situation is unique to monochorionic twins, and we discuss the implications of this for intrauterine therapy of twin-to-twin transfusion syndrome.
Assuntos
Aborto Induzido , Doenças em Gêmeos , Transfusão Feto-Fetal/terapia , Adulto , Líquido Amniótico/metabolismo , Feminino , Retardo do Crescimento Fetal/complicações , Transfusão Feto-Fetal/complicações , Transfusão Feto-Fetal/patologia , Humanos , Hidropisia Fetal/complicações , Placenta/patologia , Poli-Hidrâmnios/complicações , GravidezRESUMO
Two cases of tumors caused by osteoblastoma in children less than 3 years old are presented. This report points out that although most osteoblastomas appear in patients less than 30 years old, it is uncommon for the lesion to present in the very young. The authors note that osteoblastoma should be considered in the differential diagnosis of a lytic lesion because patients may be too young to communicate their complaints.
Assuntos
Osteoma Osteoide/diagnóstico , Neoplasias da Medula Espinal/diagnóstico , Pré-Escolar , Diagnóstico Diferencial , Humanos , Recém-Nascido , Masculino , Osteoma Osteoide/diagnóstico por imagem , Osteoma Osteoide/patologia , Osteoma Osteoide/cirurgia , Radiografia , Neoplasias da Medula Espinal/diagnóstico por imagem , Neoplasias da Medula Espinal/patologia , Neoplasias da Medula Espinal/cirurgiaRESUMO
Selective embryocide was performed as a two-stage procedure in a patient with a quintuplet pregnancy in the first trimester. No complications occurred, and the patient was delivered of healthy twins at term. This procedure may be offered to selected patients with pregnancies with greater than five embryos.
Assuntos
Aborto Induzido , Gravidez Múltipla , Quíntuplos , Adulto , Embrião de Mamíferos/efeitos dos fármacos , Feminino , Humanos , Indução da Ovulação , Gravidez , Cloreto de Sódio/administração & dosagem , Gêmeos , UltrassonografiaRESUMO
We report a case of choriocarcinoma in situ arising from a term placenta in an otherwise normal pregnancy that resulted in fetal hydrops and intrauterine fetal death from chronic fetal-maternal hemorrhage (FMH). The clinical and pathologic features are described and compared with the few similar cases reported and with an additional placental choriocarcinoma found in our files. We also describe the clinical and pathologic observations of two chorangiomas that caused massive FMH and led to fetal death.
Assuntos
Coriocarcinoma/patologia , Transfusão Feto-Materna/etiologia , Doenças Placentárias/patologia , Complicações Neoplásicas na Gravidez/patologia , Adulto , Coriocarcinoma/complicações , Coriocarcinoma/ultraestrutura , Feminino , Hemangioma/patologia , Hemangioma/ultraestrutura , Humanos , Necrose , Placenta/patologia , Doenças Placentárias/complicações , GravidezRESUMO
We describe the prenatal diagnosis of a fetus at risk for mucopolysaccharidosis (MPS) Type IVA (Morquio syndrome) using enzyme analysis of chorionic villus tissue. The family had two previous affected children, one with progressive nonimmune hydrops fetalis presenting at 16 weeks gestation and one mildly affected 5 year old. The parents had decreased levels of N-acetyl galactosamine-6-sulphate sulphatase in cultured skin fibroblasts indicating that carrier detection is possible for Morquio A syndrome.
Assuntos
Vilosidades Coriônicas/enzimologia , Edema/etiologia , Doenças Fetais/diagnóstico , Mucopolissacaridose IV/diagnóstico , Diagnóstico Pré-Natal , Adulto , Pré-Escolar , Condroitina Sulfatases/metabolismo , Feminino , Idade Gestacional , Humanos , Mucopolissacaridoses , Gravidez , Pele/enzimologiaRESUMO
Twin transfusion syndrome is a serious complication of monozygotic twin pregnancy. Diagnostic ultrasound now allows early diagnosis of this condition, but therapy has remained more elusive. In this article we present a case of severe twin transfusion syndrome diagnosed early in the second trimester. At 25 weeks' gestation, severe hydramnios, premature labor, and growth retardation of the donor twin suggested that selective feticide be contemplated to allow continuation of the pregnancy for the remaining twin. This was accomplished successfully by an in utero approach with subsequent follow-up and delivery of a healthy female infant at 37 weeks' gestation. Discussion of diagnosis, management, follow-up, and pathologic features is provided.
Assuntos
Morte Fetal/induzido quimicamente , Transfusão Feto-Fetal/terapia , Solução Salina Hipertônica/administração & dosagem , Cloreto de Sódio/administração & dosagem , Adulto , Feminino , Monitorização Fetal , Humanos , Injeções/métodos , Gravidez , UltrassonografiaRESUMO
A brother and sister from consecutive pregnancies had rigid and tightly adherent skin in association with generalized contractures, unusual facies, pulmonary hypoplasia, an abnormal placenta, and a short umbilical cord. Both died shortly after birth. Pathologic examination of the skin by light and electron microscopy showed structural abnormalities of the epidermis, dermis, and subcutaneous fat. An abnormal pattern of keratin proteins was determined biochemically using extracted epidermal proteins. Autopsy showed a normal spinal cord and muscle histology. It is postulated that the defective skin severely restricted movement and secondarily led to the other abnormalities. Familial occurrence is most consistent with autosomal recessive transmission. These patients and the primary skin defect are discussed within the framework of the Fetal Akinesia or Hypokinesia Deformation Sequence.