RESUMEN
The generally accepted definition of ecthyma gangrenosum (EG) states that this condition is pathognomonic of Pseudomonas septicemia (Pseudomonas aeruginosa) and that it should usually be seen in immunocompromised patients, particularly those with underlying malignant disease. The cases described in the literature present a somewhat different picture. Our objective was to analyze this controversy. The review analyzes 167 cases of EG that were described in the literature from 1975 to 2014. All articles on EG cases with EG-specific tissue defect that had signs of general and/or local infection and skin necrosis were included and analyzed, whatever the etiology detected. Necrotic lesions of the skin diagnosed as EG have various microbiological etiology, can occur in immunocompetent or even healthy persons, and are not necessarily connected with septicemia. In published cases, P. aeruginosa was detected in 123 cases (73.65%); of them, there were only 72 cases (58.5%) with sepsis. Other bacterial etiology was detected in 29 cases (17.35%) and fungi were detected in 15 cases (9%). While the clinical picture of the disease and the treatment strategy remain the same, there is no need to invent two separate definitions for Pseudomonas and non-Pseudomonas cases. We suggest accepting a broader definition of EG.
Asunto(s)
Bacterias/aislamiento & purificación , Infecciones Bacterianas/epidemiología , Ectima/epidemiología , Ectima/patología , Hongos/aislamiento & purificación , Micosis/epidemiología , Bacterias/clasificación , Infecciones Bacterianas/microbiología , Infecciones Bacterianas/patología , Hongos/clasificación , Humanos , Micosis/microbiología , Micosis/patologíaRESUMEN
OBJECTIVE: To investigate the role of Pantoea agglomerans as an infectious agent that causes infection in a wound even after the wound was managed at the emergency department. METHOD: A retrospective cohort study, reviewing the medical records of patients with traumatic wounds that were admitted to the emergency department from 2007-20 12 and had signs of wound infection for more than I 0 days after the wound was managed. Bacteriological results, clinical picture,and treatment results were obtained. RESULTS: Nine cases were identified. Pantoea agglomerans was detected in all cases. After 1-2 months of ineffective treatment, patients were hospitalised and surgical revisions of the wounds were performed.In all cases, small foreign bodies of plant origin were detected. After surgical revision, wounds were healed in 2-3 days. CONCLUSION: In cases of prolonged healing of post-traumatic wounds, the presence of foreign bodies of plant origin infected with Pantoea agglomerans should be taken into account. Removal of such foreign bodies leads to rapid healing of the wounds.
Asunto(s)
Infecciones por Enterobacteriaceae/microbiología , Cuerpos Extraños/microbiología , Pantoea/aislamiento & purificación , Plantas/microbiología , Infección de Heridas/microbiología , Heridas Penetrantes/microbiología , Adolescente , Adulto , Niño , Terapia Combinada , Infecciones por Enterobacteriaceae/terapia , Femenino , Cuerpos Extraños/terapia , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Cicatrización de Heridas , Infección de Heridas/terapia , Heridas Penetrantes/terapiaRESUMEN
OBJECTIVE: Endoscopic endonasal Draf II frontal sinusotomy is indicated for a variety of pathologies such as mucocele and non-responsive chronic frontal sinusitis. However, this approach is challenged and controversial. The objectives were to evaluate the advantages, disadvantages, indications, and rate of complications of this approach, without the use of a navigation system. METHODS: The files and computed tomography (CT) scans of 25 patients who underwent endoscopic endonasal Draf II sinusotomy at Assaf Harofeh Medical Center between 1999 and 2002 were reviewed. RESULTS: Thirty-one frontal sinuses were operated on and follow-up was between 18 and 62 months (average 30.3). Twenty-two sinuses (71%) had previous surgery. The Draf II procedure was used in 3.7% of all cases during the survey period. The most frequent indication for surgery was inflammation (48%) followed by mucocele (28%). In all but 2 sinuses (93%), the frontal floor between the lamina papyracea and the middle concha was drilled out. Twenty-four patients (96%) were successfully ventilated. No major complications were noted. CONCLUSIONS: The Draf II approach can be used safely and successfully without a navigation system, including cases of revision endoscopic sinus surgery. Correct interpretation of the surgical field and a CT scan are crucial for success. Careful patient selection is essential for this procedure.
Asunto(s)
Endoscopía/métodos , Seno Frontal/cirugía , Enfermedades de los Senos Paranasales/cirugía , Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
A segment comprising 307,078 nucleotides of the pig major histocompatibility complex (SLA) was completely sequenced. The segment corresponded to the entire SLA classical class I-containing region of the serologically defined SLA H01 haplotype. In all, 11 genes were characterized, comprising 7 class I genes located on the centromeric part of the sequence (SLA-1, 2, 3, 4, 5, 9, and 11) and 4 ring finger-related family genes located on its telomeric part. No member of one family was intermingled with a member of the other or with any third-party gene. All class I genes except SLA-11 were similarly orientated. The SLA-1, 2, and 3 genes displayed both promoter and overall coding regions compatible with normal functions. The SLA-4, 11, and 9 genes were considered pseudogenes because they exhibited marked anomalies. Although the SLA-5 gene had a complete coding region, it displayed mutations in promoter elements which could modify its expression. The great molecular similarity observed among the class I genes extended far outside them, and resulted from segmental duplications. The ring finger genes exhibited great homology with their human counterparts. In pig, one of these genes appeared to correspond to a complete gene which in humans is probably a pseudogene. In all, the 11 genes characterized span about 20% of the total sequence. The remaining 80% consists of interspersed repeat elements. The present results, together with the sequence previously reported involving the SLA class I-related genes, open the way for a better understanding of pig MHC organization.
Asunto(s)
Genes MHC Clase I , Porcinos/genética , Alelos , Animales , Secuencia de Bases , Centrómero/genética , Proteínas de Unión al ADN/genética , Haplotipos , Antígenos de Histocompatibilidad Clase I/inmunología , Prueba de Histocompatibilidad , Humanos , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Homología de Secuencia de Ácido Nucleico , Telómero/genética , Regiones no Traducidas , Dedos de ZincRESUMEN
A segment of 158,063 nucleotides of the pig major histocompatibility complex (SLA) and corresponding to the junction of the class I and class III regions was sequenced entirely. The centromeric part of the segment contained six class III genes including the three tumor necrosis factor genes, while the telomeric part contained three genes belonging to the class I region. The order and the molecular organization of these genes were exactly conserved in the SLA and HLA complexes, except for the SC1 gene which displayed a shift of the reading frame in swine. The cluster of the three SLA class I-related genes (Ib) and the MIC1 and MIC2 genes were located in the middle of the segment, in the following order from the centromeric side onwards, SLA-6, SLA-7, SLA-8, MIC-1 and MIC-2. All three SLA Ib genes displayed an overall molecular structure compatible with the expression of membrane-anchored glycoproteins. The SLA-7 and SLA-8 genes bear greater resemblance than to the SLA-6 gene. Six SLA-6 alleles have been previously defined differing each from the other by unique point mutations. One of them, appeared to have arisen through the occurrence of a gene conversion event in which the SLA-7 gene served as template. Only MIC-2 gene might be functional, the second MIC-1 gene being truncated. In all, the 14 genes characterized spans 37% of the total sequence. The remaining 63% nucleotides comprised a number of repeat DNA motives, including LINE fragments, SINEs, microsatellites, and also numerous nucleotide stretches not yet defined in swine.
Asunto(s)
Genes MHC Clase I/genética , Complejo Mayor de Histocompatibilidad/genética , Porcinos/genética , Alelos , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Mapeo Cromosómico/métodos , Orden Génico/genética , Antígenos de Histocompatibilidad Clase I/química , Antígenos de Histocompatibilidad Clase I/genética , Antígenos de Histocompatibilidad Clase I/aislamiento & purificación , Humanos , Masculino , Datos de Secuencia Molecular , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido NucleicoRESUMEN
The physical alignment of the entire region of the pig major histocompatibility complex (MHC) has been almost completed. In swine, the MHC is called the SLA (swine leukocyte antigen) and most of its class I region has been sequenced. Over one hundred genes have been characterised, including the classical class I and class I-related genes, as well as the class II gene families. These results in swine provide new evidence for the striking conservation during the evolution of a general MHC framework, and are consistent with the location of the class I genes on segments referred to as permissive places within the MHC class I region. Recent results confirm the involvement of the SLA region in numerous quantitative traits.
RESUMEN
To evaluate the regulation of plasma von Willebrand factor (vWF) and its in situ production by endothelial cells (ECs), 12 swine leukocyte antigen (SLA)-compatible left lung transplantations were performed. Normal lungs were transplanted into 10 pigs homozygous for von Willebrand disease and into 2 normal pigs. Additionally, 1 normal pig underwent pneumonectomy, and 1 SLA-incompatible lung transplantation between normal pigs was performed. None of the transplanted animals received immunosuppressive therapy. Plasma vWF level was evaluated by ELISA and multimeric pattern. EC vWF content was assessed by immunohistochemistry. Global hemostasis was assessed by standardized ear bleeding time. Six of 12 SLA-compatible lung transplantations and the incompatible transplantation were successful and were used for the study. The functions and the viability of ECs, reflected by their ability to produce vWF and normal multimeric plasma vWF pattern, were preserved in SLA-compatible and -incompatible lung transplantations. vWF production was preserved in ECs that initially synthesized it. EC constitutive and storage pathways are modulated differently according to transplantation compatibility and severity of rejection. In SLA-compatible lung transplantations without histological evidence of rejection, the production of vWF was preserved, whereas constitutive vWF secretion appeared to be altered in cases with minor histological signs of rejection. In pigs with von Willebrand disease that were transplanted with normal lungs without sign of rejection, plasma vWF was significantly increased in an amount expected from the estimated production of a normal lung. In the transplanted normal lung, there was no vWF overexpression by the ECs and no recruitment of ECs that initially did not express vWF. In SLA-incompatible transplantation, ECs were morphologically normal with increased and blurred vWF labeling, whereas plasma vWF levels remained normal, reflecting that EC activation is associated with an increased vWF production with probable diversion to storage pathway. This model depicts the changes of EC regulation of vWF secretion in pig lung transplants. However, this model cannot be directly extrapolated to human organ transplantation because animals did not receive any immunosuppressive therapy, which may be toxic to ECs.
Asunto(s)
Trasplante de Pulmón , Alveolos Pulmonares/metabolismo , Factor de von Willebrand/biosíntesis , Factor de von Willebrand/genética , Enfermedad Aguda , Anastomosis Quirúrgica , Animales , Antígenos , Tiempo de Sangría , Endotelio/química , Endotelio/metabolismo , Expresión Génica/fisiología , Rechazo de Injerto/inmunología , Rechazo de Injerto/metabolismo , Homocigoto , Leucocitos/química , Leucocitos/inmunología , Necrosis , Fenotipo , Neumonectomía , Alveolos Pulmonares/patología , Porcinos , Insuficiencia del Tratamiento , Factor de von Willebrand/análisisRESUMEN
Bacterial artificial chromosome (BAC) clones were assigned within the pig major histocompatibility complex (Mhc) by polymerase chain reaction-screening and Southern blot hybridization using sequence-tagged site (STS) markers and BAC end-rescued sequences. In all, 35 BAC clones were discovered containing 12 anchor genes of the SLA class I region and two genes of the SLA class III region. Twenty of these 35 clones comprised two distinct class I gene clusters, each spanning about 100 kilobases. One cluster enclosed three class I related genes (SLA-6 to -8) and two genes (MIC-1 and MIC-2) more distantly related to class I. The other cluster enclosed typical class I genes, of which three (SLA-1, -2, and -3) were transcribed by fibroblasts homozygous for the H01 haplotype which we used to construct a pig BAC library. Ordered clones are certainly helpful in isolating agronomically, biologically, and medically important genes. They would also be useful for inducing genetic modifications in pig cell lines.
Asunto(s)
Genes MHC Clase I , Porcinos/genética , Animales , Secuencia de Bases , Southern Blotting , Células Cultivadas , Cromosomas Bacterianos/genética , Electroforesis en Gel de Campo Pulsado , Fibroblastos/metabolismo , Biblioteca de Genes , Vectores Genéticos/genética , Rechazo de Injerto/prevención & control , Haplotipos/genética , Humanos , Complejo Mayor de Histocompatibilidad/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Porcinos/inmunología , Porcinos Enanos/genética , Transcripción Genética , Trasplante Heterólogo/inmunologíaRESUMEN
A porcine bacterial artificial chromosome (BAC) library was constructed using the pBeloBAC11 vector. It comprised 107,520 clones with an average insert size of 135 kb, representing an almost fivefold coverage of the swine haploid genome. Screening of the library allowed recovery of one to eight clones for 142 unique markers located all over the genome, while it failed for only one marker. About 4% chimeric clones were found. The library was also screened for the protease gene of type C porcine endoviral sequences (PERVs), and 62 clones were recovered, all but two of which contained one protease gene. We found 20 protease sequences (PERV-1 to PERV-20) which, despite differing by point mutations, were all coding sequences. The most frequent sequence, PERV-2, was 100% similar to a protease sequence expressed in the porcine PK-15 cell line. Most of the clones harbored envelope genes. Thirty-three BAC clones were mapped by fluorescence in situ hybridization to 22 distinct locations on 14 chromosomes, including the X and Y chromosomes. These overall results indicate that there is generally one PERV copy per integration site. Although PERV sequences were not tandemly arranged, clusters of integration sites were observed at positions 3p1.5 and 7p1.1. Southern blot experiments revealed 20-30 PERV copies in the Large White pig genome studied here, and variations in PERV content among pigs of different breeds were observed. In conclusion, this BAC collection represents a significant contribution to the swine large genomic DNA cloned insert resources and provides the first detailed map of PERV sequences in the swine genome. This work is the first step toward identification of potential active sites of PERV elements.
Asunto(s)
Cromosomas Bacterianos/genética , Endopeptidasas/genética , Gammaretrovirus/genética , Porcinos/genética , Secuencia de Aminoácidos , Animales , Southern Blotting , Mapeo Cromosómico , ADN/genética , Biblioteca Genómica , Hibridación Fluorescente in Situ , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Proteínas Virales/genéticaRESUMEN
In swine, the major histocompatibility complex (Mhc) or swine leukocyte antigen (SLA) is located on chromosome 7 and divided by the centromere. Thus, the telomeric class I and more centromeric class III regions are located on the p arm and the class II region is located on the q arm. The SLA region spans about 2 Mb, in which more than 70 genes have so far been characterized. Despite its division by the centromere, the spatial relationships between the genes in the class II and class III regions, and between the well-conserved non-class I genes of the class I region, are similar to those found in the human HLA complex. On the other hand, no orthologous relationships have been found between the Mhc class I genes in man and swine. In swine, the 12 SLA class I sequences constitute two distinct clusters. One cluster comprises six classical class I-related sequences, while the other comprises five class I-distantly related sequences including two swine homologous genes of the HLA Mhc class I chain-related gene (MIC) sequence family. The number of functional SLA classical class I genes, as defined by serology, probably varies from one to four, depending on the haplotype. Some of the SLA class I-distantly related sequences are clearly transcribed. As regards the SLA class II genes, some of them clearly code for at least one functional SLA-DR and one SLA-DQ heterodimer product, but none code for any DP product. The amino acid alignment of the variable domains of 33 SLA classical class I chains, and 62 DR beta and 20 DQ beta chains confirmed the exceptionally polymorphic pattern of these polypeptides. Among the class II genes, the genes are either monomorphic, like the DRA gene, or oligomorphic, like the DQA genes. In contrast, the DRB and DQB genes display considerable polymorphism, which seems more marked in DRB than DQB genes.
Asunto(s)
Complejo Mayor de Histocompatibilidad , Porcinos/genética , Porcinos/inmunología , Secuencia de Aminoácidos , Animales , Humanos , Datos de Secuencia MolecularRESUMEN
To evaluate the relative role of plasma and platelet von Willebrand factor (vWF) pools in hemostasis and arterial thrombogenesis, pigs with vW disease (vWD) were injected with vWF concentrate and/or grafted with bone marrow from a normal pig. Hemostasis was assessed by measurement of ear immersion bleeding time, factor VIII (FVIII) activity, and plasma and platelet vWF antigen levels. The thrombotic process was explored at 650 s(-1) and 1600 s(-1) in an ex vivo cylindrical perfusion chamber. Pigs with vWD exhibited a prolonged bleeding time (>30 minutes) compared with normal pigs (<5 minutes); in addition, they showed normal platelet adhesion and thrombus formation at 650 s(-1) but profoundly reduced platelet adhesion and thrombus formation at 1600 s(-1). Each experiment was performed before and 3 and 24 hours after injection of vWF concentrate. In our bleeding time study, only plasma vWF restoration induced a partial but delayed correction (24 hours postinjection), which was correlated with the highest measured level of FVIII activity. In the perfusion chamber model, restoration of plasma or platelet vWF pools resulted in similar partial correction of platelet adhesion and average thrombus size. In the perfused pigs, the maximum correction occurred 3 hours postinjection. Platelet deposition reached normal values after vWF concentrate was injected into a grafted pig. The present results suggest that when both plasma and platelet vWF levels are restored in vWD pigs, bleeding time and the thrombotic process are normalized according to different kinetics and with differing degrees of effectiveness.
Asunto(s)
Plaquetas/fisiología , Hemostasis/fisiología , Trombosis/fisiopatología , Factor de von Willebrand/fisiología , Animales , Tiempo de Sangría , Susceptibilidad a Enfermedades , Perfusión , Adhesividad Plaquetaria/fisiología , Estrés Mecánico , PorcinosRESUMEN
The major histocompatibility complex in swine (swine leucocyte antigen: SLA) is located on chromosome 7 with the class I and class III regions separated by the centromere from the class II region. The overall molecular organisation of the class I and III regions is well known, but further research is needed to establish that of the class II region. Approximately sixty genes have been characterised to date, including ten tightly packed SLA class I sequences. The exact number of functional polymorphic class I genes, as defined by serology, probably varies from one to four, depending on the haplotype. At least two other distantly class I-related gene families exist. The numerous and significant associations reported between SLA haplotypes and physiological traits are described. These traits include immune responsiveness to a variety of microbes and metazoan parasites, and male and female production and reproduction performance. The results obtained suggest that selection for specific SLA haplotypes may assist in the improvement of porcine production.
Asunto(s)
Antígenos de Histocompatibilidad/genética , Complejo Mayor de Histocompatibilidad , Porcinos/inmunología , Animales , Mapeo Cromosómico , Haplotipos , Complejo Mayor de Histocompatibilidad/genética , Porcinos/genéticaRESUMEN
Fibrosis is characterized by proliferation of fibroblasts and deposition of extracellular matrix (ECM). As alterations in the composition of ECM may account for its chronic extension, we studied the expression of the tenascin-C (TN-C) and tenascin-X (TN-X) ECM glycoproteins in our pig model of the effects of accidental exposures to radiation, in which cutaneous and muscle fibrosis developed after the induction of necrosis after a high single dose (160 Gy at the skin surface) of gamma rays. We found that, in the healed fibrotic dermis and underlying muscle fibrosis, the amount of TN-C mRNA was increased up to 18- and 39-fold, respectively, compared to normal dermis, whereas the level of TN-X mRNA remained almost unchanged. In analyses by Western blotting, the two main TN-C isoforms of 235-240 and 190-200 kDa increased up to 45- and 105-fold in fibrotic tissues, respectively. The large isoform was expressed more strongly than the smaller, although in healed fibrotic scar tissues their ratio was lower in protein than in RNA. Compared to unirradiated skin, an immunohistological study revealed stronger TN-C staining at the dermo-epidermal junction and in areas of remodeling in healed skin. An intense extracellular staining was observed around myofibroblasts in muscle fibrosis. Therefore, the gene encoding TN-C is highly up-regulated in fibrotic tissues, and mechanisms regulating the levels of TN-C variants occur at both the RNA and protein levels. Each isoform might play a distinct role in the chronic activation of fibrosis by differentially regulating mechanisms like cell adhesion, migration or proliferation.
Asunto(s)
Rayos gamma , Músculos/efectos de la radiación , Piel/efectos de la radiación , Tenascina/genética , Regulación hacia Arriba , Empalme Alternativo , Animales , Femenino , Fibrosis , Inmunohistoquímica , Músculos/metabolismo , Músculos/patología , Necrosis , ARN Mensajero/genética , ARN Mensajero/metabolismo , Dosis de Radiación , Piel/metabolismo , Piel/patología , Porcinos , Tenascina/biosíntesisRESUMEN
A map of the SLA complex, or swine major histocompatibility complex (MHC), class I region was constructed by alignment of yeast artificial chromosomes (YACs) harboring MHC class I genes as well as anchor genes already mapped within the human MHC complex (HLA). Five YACs containing 9 anchor genes built a contig of about 1.0-1.2 Mb between the SLA class III BAT1 locus and the olfactory receptor-like genes OLF42. Ten different SLA class I sequences, including putative allelic forms of published classical and non-classical SLA class I genes, were assigned to the 400-kb enclosing centromeric part of the contig. Three additional YACs comprising the OLF89 genes and two YACs containing the butyrophilin gene were located telomeric to the contig. Comparison between the human and porcine MHC complexes showed a perfect conserved order of anchor genes, whereas no orthologous relationships were found for the class I loci.
Asunto(s)
Mapeo Cromosómico , Genes MHC Clase I , Porcinos/genética , Porcinos/inmunología , Animales , Secuencia de Bases , Cromosomas Artificiales de Levadura/genética , ADN/genética , Cartilla de ADN/genética , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Ácido Nucleico , Especificidad de la EspecieRESUMEN
A swine DNA genomic library was constructed in yeast artificial chromosome (YAC) using the pYAC4 vector and the AB1380 strain. The DNA prepared from two Large White males was partially digested with EcoRI and size selected after both digestion and ligation. The YAC library contained 33792 arrayed clones with an average size of 280 kb as estimated by analysis of 2% of the clones, thus representing a threefold coverage of the swine haploid genome. The library was organized in pools to facilitate the PCR screening. The complexity of the library was tested both for unique and centromeric repeated sequences. In all, 20 out of 22 primer sets allowed the characterization of one to six clones containing specific unique sequences. These sequences are known to be on Chromosomes (Chrs) 1, 2, 5, 6, 7, 8, 13, 14, 15, 17, and X. Eight additional clones carrying centromeric repeat units were also isolated with a single primer set. The sequencing of 37 distinct repeat units of about 340 bp subcloned from these eight YACs revealed high sequence diversity indicating the existence of numerous centromeric repeat unit subfamilies in swine. Furthermore, the analysis of the restriction patterns with selected enzymes suggested a higher order organization of the repeat units. According to preliminary FISH experiments on a small number of randomly chosen YACs and YACs carrying specific sequences, the chimerism appeared to be low. In addition, primed in situ labeling experiments favored the idea that the YACs with centromeric repeat sequences were derived from a subset of metacentric and submetacentric chromosomes.
Asunto(s)
Centrómero , Cromosomas Artificiales de Levadura , Biblioteca Genómica , Secuencias Repetitivas de Ácidos Nucleicos , Porcinos/genética , Animales , Secuencia de Bases , Mapeo Cromosómico , ADN , Hibridación Fluorescente in Situ , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Saccharomyces cerevisiae/genéticaRESUMEN
In swine, distinct centromeric satellite DNA families have been described that correspond to either all the metacentric chromosomes except the Y (Mc1) or all the acrocentric chromosomes (Ac2). Using primed in situ (PRINS) labeling, we show here that primers derived from various sequences specifically label the centromeres of different subgroups of chromosomes. Among five primers derived from centromeric sequences of acrocentric chromosomes reported to be very homogeneous, four recognize all the acrocentric chromosomes, whereas one labels prominently chromosome 17. For the metacentric chromosomes, six primers have been derived from several divergent sequences. Among these primers, two recognize all the metacentric chromosomes except 5, 10, and 12. Three other primers label small subsets of metacentric chromosomes, including the X and one or two additional chromosomes. The last primer is specific to chromosome 1. These preliminary results suggest that it should be possible to define specific primers for almost every swine chromosome. Already, some of the primers reported here permit a distinction between swine chromosomes difficult to differentiate without banding, such as the X chromosome and chromosome 9. Therefore, the PRINS technique using centromeric motifs constitutes an additional tool for cytogenetic studies in swine.
Asunto(s)
Centrómero/genética , ADN Satélite/genética , Secuencias Repetitivas de Ácidos Nucleicos , Porcinos/genética , Animales , Secuencia de Bases , Mapeo Cromosómico , Secuencia de Consenso , Hibridación Fluorescente in Situ , Datos de Secuencia MolecularRESUMEN
A highly significant genetic association has been found between some alleles of the swine Major Histocompatibility Complex SLA (Swine Leukocyte Antigen genetic complex) and the cytosolic malic enzymatic activity level in muscles. The aim of this study was to find out whether this genetic association was due to a close linkage of the SLA region and the gene coding for the enzyme. Since no swine cytosolic malic enzyme sequence (ME1) was available, we isolated several overlapping fragments that spanned the almost entire malic enzyme transcript both by screening of a swine cDNA library and by RT-PCR. The results indicated the existence of two transcripts of 2. 0 and 3.1 kb, which probably correspond to two alternative forms of one gene. The sequence of the transcript was highly similar to the other published mammalian cytosolic NADP+-dependent malic enzyme cDNA, especially within the four functional domains. Two major bands at 3.7 and 2.4 kb were detected on Northern blots containing the RNA from 25 tissues from fetuses and adult pigs. A high expression level was found in the adrenal gland, muscle, liver, and peripheral nerves. The analysis of malic enzyme RFLPs in five SLA informative families revealed an independent segregation of the ME1 gene from the SLA region. In situ hybridization results localized the cytosolic malic enzyme on the swine Chromosome (Chr) 1p1.2, except that the association between SLA and the malic enzyme activity level was due to a physical genetic linkage. Thus, the mechanisms underlying this association remain to be elucidated.