RESUMO
AIMS: Cytoplasmic accumulation of the nuclear protein transactive response DNA-binding protein 43 (TDP-43) is an early determinant of motor neuron degeneration in most amyotrophic lateral sclerosis (ALS) cases. We previously disclosed this accumulation in circulating lymphomonocytes (CLM) of ALS patients with mutant TARDBP, the TDP-43-coding gene, as well as of a healthy individual carrying the parental TARDBP mutation. Here, we investigate TDP-43 subcellular localization in CLM and in the constituent cells, lymphocytes and monocytes, of patients with various ALS-linked mutant genes. METHODS: TDP-43 subcellular localization was analysed with western immunoblotting and immunocytofluorescence in CLM of healthy controls (n = 10), patients with mutant TARDBP (n = 4, 1 homozygous), valosin-containing protein (VCP; n = 2), fused in sarcoma/translocated in liposarcoma (FUS; n = 2), Cu/Zn superoxide dismutase 1 (SOD1; n = 6), chromosome 9 open reading frame 72 (C9ORF72; n = 4), without mutations (n = 5) and neurologically unaffected subjects with mutant TARDBP (n = 2). RESULTS: TDP-43 cytoplasmic accumulation was found (P < 0.05 vs. controls) in CLM of patients with mutant TARDBP or VCP, but not FUS, in line with TDP-43 subcellular localization described for motor neurons of corresponding groups. Accumulation also characterized CLM of the healthy individuals with mutant TARDBP and of some patients with mutant SOD1 or C9ORF72. In 5 patients, belonging to categories described to carry TDP-43 mislocalization in motor neurons (3 C9ORF72, 1 TARDBP and 1 without mutations), TDP-43 cytoplasmic accumulation was not detected in CLM or in lymphocytes but was in monocytes. CONCLUSIONS: In ALS forms characterized by TDP-43 mislocalization in motor neurons, monocytes display this alteration, even when not manifest in CLM. Monocytes may be used to support diagnosis, as well as to identify subjects at risk, of ALS and to develop/monitor targeted treatments.
Assuntos
Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Monócitos/metabolismo , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , MutaçãoRESUMO
AIM: Between 15-30% of patients presenting with low back pain have some SI joint involvement. The diagnosis of SI joint involvement in low back pain is quite difficult and depends on a detailed combination of clinical manoeuvres and injection tests. In 5% of patients with SI joint pain, the joint is physically unstable (termed disruption) resulting in ineffective medical and conservative therapeutic options. In this study we present the results of the first 12 cases of SI joint disruption treated using a minimally invasive SI joint arthrodesis system in order to evaluate the safety and the efficacy of this system. METHODS: Medical charts at a single center were reviewed for demographics, perioperative metrics, patient reported outcomes for pain, function and quality of life (NRS, ODI and RDQ respectively), as well as satisfaction with surgery (yes/no) and results of postoperative CT scan. RESULTS: Mean age was 53 years (range 36-71) and all patients were female. Patient reported outcomes at follow up (range 8-18 months) improved clinically as well as statistically as evidenced by a mean improvement in pain on NRS of 4 points, back related function on ODI by 19.4 points, and in quality of life measured using RDQ of 13.6 points (all P=0.01). Local hematoma requiring drainage was apparent in 2 patients. Patient satisfaction was 100%. All 3 month CT scans showed initial fusion. CONCLUSION: The results of this study confirm that MIS SI joint fusion using the iFuse Implant System is safe and effective method of treating patients with SI joint disruption.
Assuntos
Artrodese/métodos , Dor Lombar/cirurgia , Articulação Sacroilíaca/cirurgia , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Satisfação do Paciente , Projetos Piloto , Qualidade de Vida , Resultado do TratamentoRESUMO
PURPOSE: To evaluate granulocyte colony-stimulating factor (G-CSF) efficacy in accelerating bone regeneration following opening-wedge high tibial valgus osteotomy for genu varum. METHODS: A phase II trial was conducted for evaluating the preoperative administration of G-CSF given at 10 µg/kg/day for 3 consecutive days with an additional half-dose 4 h before the opening-wedge high tibial valgus osteotomy. Overall, 12 patients (Group A) received G-CSF treatment, and the subsequent 12 patients (Group B) underwent surgery without G-CSF. The osteotomy gap was filled by a bone graft substitute. Bone marrow cell (BMC) mobilization was monitored by CD34+ve cell and clonogenic progenitor cell analysis. All patients underwent a clinical (Lysholm Knee Scale and SF-36) and radiographic evaluation preoperatively, as well as at given intervals postsurgery. RESULTS: All patients completed the treatment program without major side effects; G-CSF was well tolerated. BMC mobilization occurred in all Group A patients, with median peak values of circulating CD34+ve cells of 110/µL (range 29-256). Circulating clonogenic progenitors paralleled CD34+ve cell levels. A significant improvement in Lysholm Knee Scale was recorded at follow-up in Group A compared to Group B. At the radiographic evaluation, there was a significant increase in osseointegration at the bone-graft junction in Group A at 1, 2, 3 and 6 months postsurgery compared to Group B. The computerized tomography scan of the grafted area at 2 months postsurgery showed no significant difference in the quality of the newly formed bone between the two Groups. CONCLUSIONS: Although the limited number of patients does not allow firm conclusions, the study suggests that G-CSF can be safely administered preoperatively in subjects undergoing opening-wedge high tibial valgus osteotomy; in addition, the clinical, radiographic and CT monitoring indicate that G-CSF and/or mobilized BMCs may hasten bone graft substitute osseointegration. LEVEL OF EVIDENCE: I.
Assuntos
Substitutos Ósseos , Genu Varum/cirurgia , Fator Estimulador de Colônias de Granulócitos/farmacologia , Mobilização de Células-Tronco Hematopoéticas/métodos , Osseointegração/efeitos dos fármacos , Osteotomia/métodos , Tíbia/cirurgia , Adulto , Idoso , Esquema de Medicação , Estudos de Viabilidade , Feminino , Seguimentos , Genu Varum/complicações , Genu Varum/diagnóstico por imagem , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Indicadores Básicos de Saúde , Humanos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/efeitos dos fármacos , Articulação do Joelho/fisiologia , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/complicações , Osteoartrite do Joelho/diagnóstico por imagem , Osteoartrite do Joelho/cirurgia , Osteotomia/instrumentação , Cuidados Pré-Operatórios , Estudos Prospectivos , Tíbia/diagnóstico por imagem , Tíbia/efeitos dos fármacos , Tíbia/fisiologia , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
The aim of this study was to assess the efficacy and safety of deslorelin acetate implants on domestic queen puberty postponement. Thirty, 114.4 ± 12.7 days old, 1.5 ± 0.1 kg prepubertal crossbred female cats were included in this study. The animals were kept under a positive photoperiod and randomly assigned to deslorelin acetate 4.7 mg SC implants (n = 15) or to a non-treated control group (n = 15). The queens were followed up daily and weighed weekly until puberty. Vaginal cytology was also carried out three times a week. Puberty was diagnosed by the presence of the typical oestrous behaviour and vaginal cytology findings. At puberty, ovariectomy was performed and the gonads grossly described. Age (281.2 ± 21.6 vs 177.8 ± 10.8; p < 0.01) but not weight (2.6 ± 0.1 vs 2.5 ± 0.1; p > 0.1) at puberty differed between the deslorelin and control groups, respectively. One deslorelin-treated female showed an oestrous response and another showed clinical signs of pyometra after the implants. Deslorelin-treated ovaries appeared small, while control gonads were normal. It was concluded that long-term-release deslorelin, administered at approximately 50% adult body weight, postponed feline puberty without altering growing rate.
Assuntos
Gatos/crescimento & desenvolvimento , Hormônio Liberador de Gonadotropina/agonistas , Maturidade Sexual/efeitos dos fármacos , Pamoato de Triptorrelina/análogos & derivados , Animais , Preparações de Ação Retardada , Implantes de Medicamento , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/farmacologia , Feminino , Pamoato de Triptorrelina/administração & dosagem , Pamoato de Triptorrelina/farmacologiaRESUMO
Antimicrobial peptides are effector molecules of innate immunity that provide a first line of defense against pathogens. In mammals, they are stored in granules of leukocytes and are present in those sites that are exposed to microbial invasion, such as mucosal surfaces and skin. In the last decade, biochemical investigations and recombinant DNA technology have allowed the identification and characterization of several antimicrobial peptides from various animal and vegetal species. Most of the mammalian peptides have been grouped in two broad families: defensins and cathelicidin-derived peptides. Functional studies have shown that the toxicity mechanisms for many peptides consist of a rapid permeabilization of the target cell membrane. In addition to their microbicidal activity, some members of both families are multifunctional molecules, playing a modulating role in the inflammation and the antigen-driven immune response.
Assuntos
Peptídeos Catiônicos Antimicrobianos/fisiologia , Imunidade Inata , Leucócitos/imunologia , Animais , Ânions/química , Peptídeos Catiônicos Antimicrobianos/classificação , Bactérias/química , Catelicidinas , Permeabilidade da Membrana Celular/efeitos dos fármacos , Grânulos Citoplasmáticos/metabolismo , Citotoxicidade Imunológica , Defensinas/fisiologia , Desenho de Fármacos , Humanos , Inflamação/metabolismo , Leucócitos/metabolismo , Mamíferos/imunologia , Mamíferos/metabolismo , Lipídeos de Membrana/química , Camundongos , Fagossomos/fisiologia , Fosfolipídeos/química , Estrutura Terciária de Proteína , Coelhos , Ratos , Células Tumorais CultivadasRESUMO
Fibronectin (FN) polymorphism is caused by alternative splicing patterns in at least three regions (ED-A, ED-B and IIICS) of the primary transcript of a single gene. Using monoclonal antibodies, we previously demonstrated that transforming growth factor-beta (TGF-beta) preferentially increases the accumulation of the FN isoforms containing the ED-A sequence in cultured normal human fibroblasts [Balza et al., (1988) FEBS Lett. 228, 42-44]. To determine the basis of this effect, we have examined through S1 nuclease analysis, the levels of ED-A- and ED-B-containing mRNAs in cultured normal human skin fibroblasts before and after TGF-beta treatment. These experiments have shown that TGF-beta increases the relative amount of m-RNA for ED-A- and ED-B-containing FN isoforms. These data demonstrate that a growth factor may regulate the splicing pattern of a pre-mRNA.
Assuntos
Fibronectinas/genética , Linfotoxina-alfa/farmacologia , Precursores de RNA/genética , Splicing de RNA , RNA Mensageiro/genética , Linhagem Celular , Sondas de DNA , Éxons , Fibroblastos/metabolismo , Humanos , Hibridização de Ácido Nucleico , Endonucleases Específicas para DNA e RNA de Cadeia SimplesRESUMO
SMAP-29 is a cathelicidin-derived peptide deduced from sheep myeloid mRNA. The C-terminally amidated form of this peptide was chemically synthesized and shown to exert a potent antimicrobial activity. Antibiotic-resistant clinical isolates highly susceptible to this peptide include MRSA and VREF isolates, that are a major worldwide problem, and mucoid Pseudomonas aeruginosa associated with chronic respiratory inflammation in CF patients. In addition, SMAP-29 is also active against fungi, including Cryptococcus neoformans isolated from immunocompromised patients. SMAP-29 causes significant morphological alterations of the bacterial surfaces, as shown by scanning electron microscopy, and is also hemolytic against human, but not sheep erythrocytes. Its potent antimicrobial activity suggests that this peptide is an excellent candidate as a lead compound for the development of novel antiinfective agents.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Proteínas Sanguíneas , Leucócitos/química , Peptídeos , Proteínas/farmacologia , Animais , Catelicidinas , Dicroísmo Circular , Relação Dose-Resposta a Droga , Desenho de Fármacos , Escherichia coli/metabolismo , Hemólise , Humanos , Cinética , Microscopia Eletrônica de Varredura , Conformação Proteica , Estrutura Secundária de Proteína , Proteínas/química , Ovinos , Staphylococcus aureus/metabolismoRESUMO
Pulmonary sarcoidosis is a disease characterized by increased numbers of T lymphocytes within the alveolar structures and the consequent spontaneous release of a variety of mediators relevant to the pathogenesis of this disorder. This phenomenon is associated with a different expansion of the T cell subpopulations present in the lung. Using monoclonal antibodies specific for T cell subsets with helper functions, we have evaluated the different T lymphocyte subpopulations present in patients with active and inactive pulmonary sarcoidosis. The small fraction of helper T cells recognized by the 5/9 monoclonal antibody appear to be preferentially expanded in the lung of patients with active disease. The functions of the 5/9+ lung T cells in pulmonary sarcoidosis were evaluated by considering the spontaneous release of monocyte chemotactic factor, the help in immunoglobulin production, and the spontaneous production of interleukin-2 by the 5/9+ lung T cells from patients with active disease. These T cell functions appeared to be restricted to the 5/9+ T cell subset. The sensitivity of the 5/9+ lung T cells to corticosteroid treatment in pulmonary sarcoidosis was studied by performing bronchoalveolar lavage in patients with active disease before and after oral prednisone therapy. Evaluation of lymphocyte subsets after 3 months of therapy showed a marked reduction of 5/9+ T cell percentages even though the overall proportion of lavage cells that were T lymphocytes was still elevated. Thus the 5/9 monoclonal antibody may be considered a good marker in gauging the activity of the alveolitis in pulmonary sarcoidosis because it recognizes the T cell subsets responsible for many activities relevant to the pathogenesis of this disorder. In addition, analysis of the proportions of 5/9+ lung T cells may result in a useful means to evaluate the early response to therapy.
Assuntos
Corticosteroides/uso terapêutico , Pneumopatias/imunologia , Pulmão/imunologia , Sarcoidose/imunologia , Linfócitos T/imunologia , Anticorpos Monoclonais/análise , Linfócitos B/imunologia , Separação Celular , Fatores Quimiotáticos/metabolismo , Humanos , Interleucina-2/análise , Pulmão/efeitos dos fármacos , Pneumopatias/tratamento farmacológico , Ativação Linfocitária , Monócitos/imunologia , Sarcoidose/tratamento farmacológicoRESUMO
Monoclonal antibodies (mAbs) against cultured human ovary carcinoma cells were produced. We obtained 7 mAbs which reacted diffusely with carcinoma of the ovary but only weakly with vessels and the surface epithelial layer of normal ovary. Biochemical characterization of these mAbs indicated that 3 out of 7 were specific for the alpha 3 chain of the Vla-3 integrin, a receptor for fibronectin, collagen and laminin. Using one of these mAbs, we have studied, by immunohistochemical methods, the distribution of alpha 3 beta 1 integrin in mucinous, serous and endometrioid cystoadenocarcinoma of the ovary and in their normal equivalent: endocervical, tubal and endometrial epithelia. The results show that alpha 3 beta 1 is present in cell-cell contact areas and more abundantly at the junction between epithelial cells and basement membrane in endocervical, tubal epithelia, in epithelium lining the cavity of the uterus and in surface epithelium of the ovary. However, endometrial glands showed only weak and fragmented positivity at the basal pole of the cells. 26 out of 31 ovarian cancers studied, expressed the alpha 3 beta 1 integrin. However, basal localization, typical of normal epithelia, is not prominent or disappears in tumors and is replaced by a more diffuse reaction with variable immunohistochemical staining of the neoplastic cells. Furthermore, a comparative analysis of the expression of alpha 3 beta 1 and its ligands, laminin (LM), fibronectin (FN) and collagen IV (Coll IV), demonstrated that basal polarization of Vla-3 was always correlated with the presence of laminin and Coll IV, intrinsic components of the basement membranes.
Assuntos
Integrinas/análise , Neoplasias Ovarianas/química , Ovário/química , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/biossíntese , Colágeno/análise , Epitélio/química , Feminino , Fibronectinas/análise , Humanos , Imuno-Histoquímica , Integrina alfa3beta1 , Laminina/análise , Camundongos , Dados de Sequência Molecular , Células Tumorais CultivadasRESUMO
Two different methods, immunofluorescence and lactoperoxidase-catalysed radioiodination, were used to investigate mIg on two different myeloma cell lines. The data indicate that, even though both techniques are highly sensitive, different results can be obtained in the same cell line depending on the technique employed.
Assuntos
Membrana Celular/análise , Imunoglobulina G/isolamento & purificação , Proteínas do Mieloma/isolamento & purificação , Receptores de Antígenos de Linfócitos B , Animais , Linhagem Celular , Células Clonais , Imunofluorescência , Radioisótopos do Iodo , Marcação por Isótopo , Lactoperoxidase , Métodos , CamundongosRESUMO
Here we describe a case of pyometra coexisting with gestation in a 4.5 year-old miniature short-haired Dachshund. The dog exhibited depression, vaginal discharge, polydipsia and dehydration. Ultrasound examination revealed the presence of low to moderate anechoic fluid collection in the left uterine horn. Blood analysis revealed mild neutrophilia with a left shift. Based on these findings a presumptive diagnosis of pyometra was made and the bitch was treated using amoxicillin-clavulanate with dopaminergic agonist (cabergoline). A second ultrasound scan revealed the presence of two gestational vesicles in the right uterine horn that were successfully carried to term. Unusually, while pyometra persisted in the left uterine horn, two viable puppies were delivered by caesarean section from the right uterine horn.
RESUMO
Interhemispheric connections have been demonstrated between the motor cortex controlling muscle pairs. However, these investigations have tended to concentrate upon hand muscles. We have extended these investigations to proximal muscles that control the scapula upon the trunk and help to move and stabilise the shoulder. Using a paired pulse transcranial magnetic stimulation protocol, the interhemispheric interactions between different shoulder girdle muscle pairs, serratus anterior, upper trapezius and lower trapezius were investigated. Test motor evoked potentials were conditioned using conditioning pulse intensities of 80% and 120% of active motor threshold at three different condition-test intervals, during three different tasks. Interhemispheric inhibition was observed in upper trapezius using a conditioning intensity of 120% and condition-test interval of 8 ms (17 ± 18%, p < 0.007). A trend towards inhibition was observed in lower trapezius and serratus anterior using a conditioning intensity of 120% and a condition-test interval of 8 ms (13 ± 22%; p < 0.07 and 10 ± 19% respectively; p < 0.07). No interhemispheric facilitation was evoked. The study demonstrates that a low level of interhemispheric inhibition rather than interhemispheric facilitation could be evoked between these muscle pairs.
Assuntos
Lateralidade Funcional/fisiologia , Córtex Motor/fisiologia , Movimento/fisiologia , Músculo Esquelético/fisiologia , Estimulação Magnética Transcraniana/métodos , Adulto , Eletromiografia , Potencial Evocado Motor/fisiologia , Feminino , Mãos/fisiologia , Voluntários Saudáveis , Humanos , Masculino , Força Muscular/fisiologia , Inibição Neural/fisiologia , Tempo de Reação/fisiologia , Escápula/fisiologia , Tórax/fisiologia , Tronco/fisiologiaRESUMO
The bovine digital vasculature contractility has been implicated in the development of laminitis. To investigate the effect of hypoxia/reoxygenation on the contractility of isolated peripheral bovine digital veins (BDVs), vessel rings were studied under isometric conditions and submitted to 30 min of hypoxia (95%N(2)-5%CO(2)) and reoxygenation (95%O(2)-5%CO(2)) conditions, respectively. The BDVs contracted with a high K(+) depolarizing solution, developed hypoxia-induced relaxation, followed by an increase in tension upon reoxygenation. In contrast, phenylephrine-contracted BDVs displayed a rapid, sustained and reversible hypoxia-induced contraction. Reoxygenation caused a rapid relaxation in phenylephrine-contracted BDVs. The presence of the endothelium did not modify the hypoxia/reoxygenation effects and hypoxia-induced contraction was still observed in a nominal Ca(2+)-free Krebs, however, the last effect was not maintained over time. The hypoxia-induced contraction in an isolated peripheral vein may contribute to the understanding of the physiology and pathophysiology of superficial venous smooth muscle contractility, particularly in the alteration of bovine digital haemodynamics under hypoxia/reoxygenation conditions.
Assuntos
Membro Anterior/irrigação sanguínea , Hipóxia/veterinária , Contração Muscular/fisiologia , Músculo Liso Vascular/fisiopatologia , Vasoconstrição/fisiologia , Veias/fisiopatologia , Animais , Cálcio/farmacologia , Bovinos , Relação Dose-Resposta a Droga , Hipóxia/fisiopatologia , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Fenilefrina/farmacologia , Vasoconstrição/efeitos dos fármacos , Vasoconstritores/farmacologia , Veias/efeitos dos fármacosRESUMO
A promising approach for musculoskeletal repair and regeneration is mesenchymal-stem-cell- (MSC-)based tissue engineering. The aim of the study was to apply a simple protocol based on mincing the umbilical cord (UC), without removing any blood vessels or using any enzymatic digestion, to rapidly obtain an adequate number of multipotent UC-MSCs. We obtained, at passage 1 (P1), a mean value of 4, 2 × 10(6) cells (SD 0,4) from each UC. At immunophenotypic characterization, cells were positive for CD73, CD90, CD105, CD44, CD29, and HLA-I and negative for CD34 and HLA-class II, with a subpopulation negative for both HLA-I and HLA-II. Newborn origin and multilineage potential toward bone, fat, cartilage, and muscle was demonstrated. Telomere length was similar to that of bone-marrow (BM) MSCs from young donors. The results suggest that simply collecting UC-MSCs at P1 from minced umbilical cord fragments allows to achieve a valuable population of cells suitable for orthopaedic tissue engineering.
RESUMO
Two experiments were conducted to investigate the effects of the GnRH antagonist acyline (330 microg/kg, given sc) on ovarian follicular development and ovulation, as well as on pregnancy maintenance in domestic cats. In the first experiment, seven queens in proestrus (total of 24 proestrus periods), were randomly assigned to treatment with either acyline (ACY; n=17) or a placebo (PLC; n=7). All queens were mated with a fertile tomcat. In the ACY and PLC groups, cessation of estrus occurred (mean+/-SEM) 7.0+/-1.3 and 7.0+/-1.7 d after treatment (P>0.1), ovulation occurred in 2 of 17 and all seven estrus periods (P<0.05), and pregnancy rates were 1 of 16 and 7 of 7 (P<0.05), respectively. In the ACY and PLC groups, intervals from treatment to the onset of the ensuing proestrus were 18.4+/-1.7 and 120+/-17.2 d. In the second experiment, 14 pregnant queens were randomly allocated, according to their mating date, to treatment with acyline in early pregnancy (from 20 to 25 d, n=3), mid pregnancy (from 26 to 45 d; n=4), late pregnancy (> 45 d; n=3), or injection of a placebo in early (n=1), mid (n=2), or late pregnancy (n=1). Ultrasonographic assessments of the uterus were done every second day for 2 wk post treatment, and serum progesterone (P(4)) concentrations were determined before treatment, and at 7 and 14 d after treatment. No pregnancies were prematurely terminated and post-treatment P(4) concentrations did not differ among treatment groups (P>0.1). In conclusion, in the domestic cat, GnRH withdrawal by acyline prevented ovulation when given in early follicular phase (proestrus), but did not significantly affect luteal function during pregnancy.
Assuntos
Gatos/fisiologia , Corpo Lúteo/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Oligopeptídeos/efeitos adversos , Folículo Ovariano/efeitos dos fármacos , Ovulação/efeitos dos fármacos , Animais , Corpo Lúteo/fisiologia , Feminino , Idade Gestacional , Masculino , Oligopeptídeos/administração & dosagem , Folículo Ovariano/crescimento & desenvolvimento , Placebos , Gravidez , Proestro , Progesterona/sangue , Ultrassonografia , Útero/diagnóstico por imagemAssuntos
Anticorpos Antivirais/análise , Vírus da Dengue/imunologia , Dengue/epidemiologia , Argentina , Feminino , Humanos , MasculinoAssuntos
Doença de Chagas/veterinária , Doenças do Cão , Animais , Argentina , Cardiomiopatia Chagásica/diagnóstico , Cardiomiopatia Chagásica/patologia , Cardiomiopatia Chagásica/veterinária , Doença de Chagas/diagnóstico , Modelos Animais de Doenças , Doenças do Cão/diagnóstico , Doenças do Cão/patologia , Cães , Eletrocardiografia/veterinária , Feminino , Masculino , Miocárdio/patologia , Testes Sorológicos/veterináriaRESUMO
Umbilical cord blood (UCB) is a source of hematopoietic progenitor cells and is used as an alternative to the bone marrow or peripheral blood for treatment of several onco-hematological diseases. Because of the limited number of CD34+ hematopoietic stem cells present in UCB units and of the elevated costs of cryopreservation, it is of paramount importance to select the UCB units that are clinically useful before storage and optimize banking efficiency by designing reliable procedures to process and freeze the selected units. Among the different parameters characterizing UCB, nucleated cell (NC) and CD34+ cell content provides useful criteria to select UCB units since clinical data documented that the infused cell load (both NC and CD34+ cells) plays an important role in the successful outcome of transplants. By evaluating volume, CD34+ cell content, NC total amount, and NC density of 117 UCB units, we found a significant association between CD34+ cell content and NC density and total amount, indicating these parameters as useful to decide UCB clinical utility. Furthermore, we set up a fast procedure to process UCB units for storage. A system for NC separation and volume reduction of UCB samples in a dedicated, germ-free, closed circuit was developed, where plasma and red blood cells (RBC) depletion was obtained by sedimentation in the presence of a 3.5% Polygeline solution. By this separation system, both RBC depletion and high NC and CD34+ cell recoveries were achieved in 60 min, and the yield was comparable to the one obtained by other separation methods. Since Polygeline has been clinically used as a plasma expander and no toxic effects on patients were reported, the protocol can be applied in the large-scale banking of UCB.