RESUMO
Retrograde autologous priming (RAP) has been routinely applied in cardiac pediatric cardiopulmonary bypass (CPB). However, this technique is performed in pediatric patients weighing more than 20 kg, and research about its application in pediatric patients weighing less than 20 kg is still scarce. This study explored the clinical application of RAP in CPB in pediatric patients undergoing cardiac surgery. Sixty pediatric patients scheduled for cardiac surgery were randomly divided into control and experimental groups. The experimental group was treated with CPB using RAP, while the control group was treated with conventional CPB (priming with suspended red blood cells, plasma and albumin). The hematocrit (Hct) and lactate (Lac) levels at different perioperative time-points, mechanical ventilation time, hospitalization duration, and intraoperative and postoperative blood usage were recorded. Results showed that Hct levels at 15 min after CPB beginning (T2) and at CPB end (T3), and number of intraoperative blood transfusions were significantly lower in the experimental group (P<0.05). There were no significant differences in CPB time, aortic blocking time, T2-Lac value or T3-Lac between the two groups (P>0.05). Postoperatively, there were no significant differences in Hct (2 h after surgery), mechanical ventilation time, intensive care unit time, or postoperative blood transfusion between two groups (P>0.05). RAP can effectively reduce the hemodilution when using less or not using any banked blood, while meeting the intraoperative perfusion conditions, and decreasing the perioperative blood transfusion volume in pediatric patients.
Assuntos
Transfusão de Sangue/estatística & dados numéricos , Procedimentos Cirúrgicos Cardíacos/métodos , Ponte Cardiopulmonar/métodos , Peso Corporal , Pré-Escolar , Feminino , Humanos , Tempo de Internação , Masculino , Período Perioperatório , Resultado do TratamentoRESUMO
This study aimed to reveal the genetic and epigenetic variations involved in a resynthesized Brassica napus (AACC) generated from a hybridization between a B. rapa (AA) landrace and B. alboglabra (CC). Amplified fragment length polymorphism (AFLP), methylation-sensitive amplified polymorphism, and the cDNA-AFLP technique were performed to detect changes between different generations at the genome, methylation, and transcription levels. We obtained 30 lines of resynthesized B. napus with a mean 1000-seed weight of over 7.50 g. All of the lines were self-compatible, probably because both parents were self-compatible. At the genome level, the S0 generation had the lowest frequency of variations (0.18%) and the S3 generation had the highest (6.07%). The main variation pattern was the elimination of amplified restriction fragments on the CC genome from the S0 to the S4 generations. At the methylation level, we found three loci that exhibited altered methylation patterns on the parental A genome; the variance rate was 1.35%. At the transcription level, we detected 43.77% reverse mutations and 37.56% deletion mutations that mainly occurred on the A and C genomes, respectively, in the S3 generation. Our results highlight the genetic variations that occur during the diploidization of resynthesized B. napus.
Assuntos
Brassica/genética , Cruzamentos Genéticos , Genoma de Planta , Poliploidia , Metilação de DNA , Testes Genéticos , Variação Genética , MutaçãoRESUMO
Retrograde autologous priming (RAP) has been routinely applied in cardiac pediatric cardiopulmonary bypass (CPB). However, this technique is performed in pediatric patients weighing more than 20 kg, and research about its application in pediatric patients weighing less than 20 kg is still scarce. This study explored the clinical application of RAP in CPB in pediatric patients undergoing cardiac surgery. Sixty pediatric patients scheduled for cardiac surgery were randomly divided into control and experimental groups. The experimental group was treated with CPB using RAP, while the control group was treated with conventional CPB (priming with suspended red blood cells, plasma and albumin). The hematocrit (Hct) and lactate (Lac) levels at different perioperative time-points, mechanical ventilation time, hospitalization duration, and intraoperative and postoperative blood usage were recorded. Results showed that Hct levels at 15 min after CPB beginning (T2) and at CPB end (T3), and number of intraoperative blood transfusions were significantly lower in the experimental group (P<0.05). There were no significant differences in CPB time, aortic blocking time, T2-Lac value or T3-Lac between the two groups (P>0.05). Postoperatively, there were no significant differences in Hct (2 h after surgery), mechanical ventilation time, intensive care unit time, or postoperative blood transfusion between two groups (P>0.05). RAP can effectively reduce the hemodilution when using less or not using any banked blood, while meeting the intraoperative perfusion conditions, and decreasing the perioperative blood transfusion volume in pediatric patients.
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Transfusão de Sangue/estatística & dados numéricos , Ponte Cardiopulmonar/métodos , Procedimentos Cirúrgicos Cardíacos/métodos , Peso Corporal , Resultado do Tratamento , Período Perioperatório , Tempo de InternaçãoRESUMO
We investigated the effect of selective cerebral ultra-profound hypothermic blood flow occlusion on brain tissue and cell metabolism to ascertain the efficacy and safety of selective deep hypothermic technologies using proton magnetic resonance spectroscopy ((1)H-MRS). The bilateral carotid artery was blocked at room temperature for 10 min. Other neck vessels were then blocked through cold perfusion of the internal carotid artery and reflux of the ipsilateral jugular vein. Thus, selective cerebral extracorporeal circulation was established. Brain temperature was reduced to 15.1° ± 0.9°C. After 60 min, cerebral blood flow recovered naturally. Routine magnetic resonance imaging (MRI), diffusion-weighted imaging (DWI), and (1)H-MRS examination of the bilateral frontal cortex and basal ganglia were performed prior to surgery and 4, 24, 72 h, 21 days after recovery. The formants and areas under the curve (AUC) of N-acetyl aspartate (NAA), choline (Cho), creatine/phosphocreatine (Cr/Cr2) were analyzed using 1H-MRS. The pre- and postoperative AUC of NAA and Cho at different time points were compared. Conventional MRI and DWI showed no abnormal signal changes in the brain parenchyma or right basal ganglia before and after surgery (P > 0.05). There was no significant difference in the ratio between NAA/(Cr+Cr2) and Cho/(Cr+Cr2) before and after surgery in the bilateral basal ganglia and frontoparietal regions of the cortex (P > 0.05). Quantitative (1)H-MRS showed that selective deep cerebral hypothermia significantly improved the brain's tolerance to ischemia and hypoxia. Our results could provide a better understanding of the efficacy and safety of selective deep hypothermia and blood flow occlusion.
Assuntos
Encéfalo/irrigação sanguínea , Encéfalo/metabolismo , Transtornos Cerebrovasculares/patologia , Espectroscopia de Prótons por Ressonância Magnética/métodos , Animais , Ácido Aspártico/análogos & derivados , Ácido Aspártico/metabolismo , Estenose das Carótidas , Transtornos Cerebrovasculares/metabolismo , Colina/metabolismo , Parada Circulatória Induzida por Hipotermia Profunda/métodos , Creatina/metabolismo , Feminino , Macaca mulatta , Masculino , RessuscitaçãoRESUMO
The single photon emission microscope (SPEM) is an instrument developed to obtain high spatial resolution single photon emission computed tomography (SPECT) images of small structures inside the mouse brain. SPEM consists of two independent imaging devices, which combine a multipinhole collimator, a high-resolution, thallium-doped cesium iodide [CsI(Tl)] columnar scintillator, a demagnifying/intensifier tube, and an electron-multiplying charge-coupling device (CCD). Collimators have 300- and 450-µm diameter pinholes on tungsten slabs, in hexagonal arrays of 19 and 7 holes. Projection data are acquired in a photon-counting strategy, where CCD frames are stored at 50 frames per second, with a radius of rotation of 35 mm and magnification factor of one. The image reconstruction software tool is based on the maximum likelihood algorithm. Our aim was to evaluate the spatial resolution and sensitivity attainable with the seven-pinhole imaging device, together with the linearity for quantification on the tomographic images, and to test the instrument in obtaining tomographic images of different mouse organs. A spatial resolution better than 500 µm and a sensitivity of 21.6 counts·s-1·MBq-1 were reached, as well as a correlation coefficient between activity and intensity better than 0.99, when imaging 99mTc sources. Images of the thyroid, heart, lungs, and bones of mice were registered using 99mTc-labeled radiopharmaceuticals in times appropriate for routine preclinical experimentation of <1 h per projection data set. Detailed experimental protocols and images of the aforementioned organs are shown. We plan to extend the instrument's field of view to fix larger animals and to combine data from both detectors to reduce the acquisition time or applied activity.
RESUMO
The single photon emission microscope (SPEM) is an instrument developed to obtain high spatial resolution single photon emission computed tomography (SPECT) images of small structures inside the mouse brain. SPEM consists of two independent imaging devices, which combine a multipinhole collimator, a high-resolution, thallium-doped cesium iodide [CsI(Tl)] columnar scintillator, a demagnifying/intensifier tube, and an electron-multiplying charge-coupling device (CCD). Collimators have 300- and 450-µm diameter pinholes on tungsten slabs, in hexagonal arrays of 19 and 7 holes. Projection data are acquired in a photon-counting strategy, where CCD frames are stored at 50 frames per second, with a radius of rotation of 35 mm and magnification factor of one. The image reconstruction software tool is based on the maximum likelihood algorithm. Our aim was to evaluate the spatial resolution and sensitivity attainable with the seven-pinhole imaging device, together with the linearity for quantification on the tomographic images, and to test the instrument in obtaining tomographic images of different mouse organs. A spatial resolution better than 500 µm and a sensitivity of 21.6 counts·s-1·MBq-1 were reached, as well as a correlation coefficient between activity and intensity better than 0.99, when imaging 99mTc sources. Images of the thyroid, heart, lungs, and bones of mice were registered using 99mTc-labeled radiopharmaceuticals in times appropriate for routine preclinical experimentation of <1 h per projection data set. Detailed experimental protocols and images of the aforementioned organs are shown. We plan to extend the instrument's field of view to fix larger animals and to combine data from both detectors to reduce the acquisition time or applied activity.
RESUMO
The peroxisome proliferator-activated receptor gamma (PPAR-γ) is a key molecule in adipocyte differentitation; it transactivates multiple target genes in lipid metabolic pathways. Using PCR-SSCP and DNA sequencing, we evaluated a potential association of an SNP (72472 G﹥T in exon7) of the bovine PPAR-γ gene with carcass and meat quality traits in 660 individuals from five Chinese indigenous cattle breeds, Qinchuan (QC), Luxi (LX), Nanyang (NY), Jiaxian (JX), and Xianan (XN). This 72472 G﹥T mutation identified a missense mutation, Q448H. Two alleles were named C and D. Allele frequencies of PPAR-γ-C/D in the five breeds were 0.7815/0.2185, 0.9/0.1, 0.7442/0.2558, 0.7051/0.2949, and 0.8333/0.1667 for QC, NY, JX, LX, and XN, respectively. Except for the XN breed, all breeds were in Hardy-Weinberg equilibrium at this locus. The polymorphism information content was low for NY and XN (0.16 and 0.24, respectively), while it was moderately high for QC, JX, and LX (0.28, 0.31 and 0.33, respectively). Correlation analysis showed significant association of this missense mutation with carcass length, backfat thickness and water holding capacity in the QC breed. Animals with the genotype CD had significantly greater carcass length than those with genotypes CC and DD, while animals with genotype CC had significantly greater backfat thickness than those with genotypes CD and DD. Animals with genotype CC had lower water holding capacity than those with the genotypes CD and DD. In conclusion, this locus is a candidate for a major quantitative trait locus affecting production traits and could be used for beef breeding selection.
Assuntos
Cruzamento , Bovinos/genética , Estudos de Associação Genética , Carne/normas , Mutação de Sentido Incorreto/genética , PPAR gama/genética , Característica Quantitativa Herdável , Sequência de Aminoácidos , Animais , Sequência de Bases , China , Éxons/genética , Frequência do Gene/genética , Loci Gênicos/genética , Genótipo , Dados de Sequência Molecular , PPAR gama/química , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples/genética , Análise de Sequência de DNARESUMO
A 1.6-kb tandem repeat sequence had previously been identified in the subtelomeric region of mini- and megabase chromosomes from Leishmania braziliensis. Southern hybridisation was used to demonstrate that the repeat is complex specific. The sequence was characterised in strains representing four species of the L. braziliensis complex. This data allowed an assessment of the evolutionary relationship of the four species. PCR primers targeted to the repeat amplify only DNA from species of the L. braziliensis complex. Titration assays indicate that a minimum of 50 fg of parasite DNA can be detected by PCR alone. Southern hybridisation increases the limit of detection to 5 fg. Interspecies variation in the repeat sequence enabled restriction enzyme digestion of PCR products to distinguish individual species within the L. braziliensis complex.
Assuntos
DNA de Protozoário/química , Leishmania braziliensis/genética , Leishmaniose Cutânea/diagnóstico , Sequências de Repetição em Tandem/genética , Telômero/genética , Animais , Sequência de Bases , Southern Blotting , Sequência Conservada , Primers do DNA/química , DNA Ribossômico/química , Leishmania braziliensis/classificação , Leishmania braziliensis/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Mapeamento por Restrição , Alinhamento de Sequência , Especificidade da EspécieRESUMO
Leishmania braziliensis M2903 contains a highly amplified small chromosome. This work is aimed at resolving its structural organization and determining whether this unusual chromosome contains specific genes encoding proteins with important functions in disease pathology or drug resistance. Our results show that the M2903 250-kb small chromosome contains LD1 sequences and has an inverted repeat structure. The LD1 sequences and two cDNAs (cDNA2 and cDNA53) were mapped on a cosmid contig, and the two cDNAs and the corresponding genomic fragments from the small chromosome were sequenced. The gene encoding cDNA2 predicts a putative GTP-binding protein with homology to other GTP-binding proteins only in the G-1 domain region; however, four other conserved motifs can be recognized. Sequence similarity to cDNA53 is located in at least five chromosomes, and its small chromosome copy is a pseudogene. An open reading frame downstream of the cDNA53 pseudogene predicts another GTP-binding protein that belongs to a new G-protein family with an unusual conserved GTP-binding domain and a newly characterized conserved sequence motif. A portion of this GTP-binding protein gene was studied previously in L. aethiopica as a recombinant antigen that reacts with human antibodies.