RESUMO
Osteoarthritis (OA) is a potentially disabling disease whose progression is dependent on several risk factors. OA management usually involves the use of non-steroidal anti-inflammatory drugs (NSAIDs) that are the primary pharmacological treatments of choice. However, NSAIDs have often been associated with unwanted side effects. Cyclooxygenase (COX)-2 specific inhibitors, such as celecoxib, have been successfully used as an alternative in the past for OA treatment and have demonstrated fewer side effects. While abundant data are available for the clinical efficacy of drugs used for OA treatment, little is known about the disease-modifying effects of these agents. A previous review published by Zweers et al. (2010) assessed the available literature between 1990 and 2010 on the disease-modifying effects of celecoxib. In the present review, we aimed to update the existing evidence and identify evolving concepts relating to the disease-modifying effects of not just celecoxib, but also other NSAIDs. We conducted a review of the literature published from 2010 to 2016 dealing with the effects, especially disease-modifying effects, of NSAIDs on cartilage, synovium, and bone in OA patients. Our results show that celecoxib was the most commonly used drug in papers that presented data on disease-modifying effects of NSAIDs. Further, these effects appeared to be mediated through the regulation of prostaglandins, cytokines, and direct changes to tissues. Additional studies should be carried out to assess the disease-modifying properties of NSAIDs in greater detail.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Osteoartrite/tratamento farmacológico , Humanos , Resultado do TratamentoRESUMO
AIM: Several procedures have been described for rectovaginal fistula with a wide range of success, but there is little information on the long-term outcome. The aim of the present study was to investigate the long-term outcome after transvaginal anterior levatorplasty (ALP) for intractable rectovaginal fistula. METHOD: Data of 16 consecutive patients undergoing transvaginal ALP with fistulectomy and closure of the rectum and vagina between 1998 and 2011 were prospectively recorded and retrospectively investigated to study the long-term outcome. RESULTS: Birth injury (n = 7), low anterior resection for rectal cancer (n = 3), pouch surgery for ulcerative colitis (n = 2) and a procedure for prolapse and haemorrhoids (n = 2) were the main causes of the fistula. Nine patients had a covering stoma before surgery. All patients underwent ALP, with a covering stoma in two patients. Infection occurred in one patient and wound rupture after surgery in another patient. These patients underwent reoperation by ALP. All fistulae had healed at a median follow-up of 84 (8-193) months after initial surgery or stoma closure. CONCLUSION: Transvaginal ALP is effective for the treatment of mid or low rectovaginal fistula. The results show that a graft is not necessary regardless of whether or not previous surgery has been performed.
Assuntos
Cirurgia Endoscópica por Orifício Natural/métodos , Procedimentos de Cirurgia Plástica/métodos , Fístula Retovaginal/cirurgia , Reto/cirurgia , Retalhos Cirúrgicos , Adulto , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Tempo , Resultado do Tratamento , VaginaRESUMO
BACKGROUND: The aim of the present study was to classify the short-term outcomes of local correction of stoma prolapse with a stapler device. METHODS: The medical records of 11 patients undergoing local correction of stoma prolapse using a stapler device were retrospectively reviewed. RESULTS: No mortality or morbidity was observed after the surgery. Median operative time was 35 min (range 15-75 min), and blood loss was minimal. Median duration of follow-up was 12 months (range 6-55 months). One of the 11 patients had a recurrent stoma prolapse. CONCLUSIONS: This technique can be a feasible, safe and minimally invasive correction procedure for stoma prolapse.
Assuntos
Neoplasias Colorretais/cirurgia , Colostomia/efeitos adversos , Grampeadores Cirúrgicos , Prolapso Visceral/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Colectomia/métodos , Doenças do Colo/cirurgia , Neoplasias Colorretais/patologia , Colostomia/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Reoperação/métodos , Estudos Retrospectivos , Medição de Risco , Fatores de Tempo , Resultado do Tratamento , Prolapso Visceral/etiologiaRESUMO
Stomal prolapse is one of the common complications in transverse colostomy and can be managed conservatively in most cases; however, laparotomy and reconstruction of the stoma may sometimes be required, especially in case of irreducible colostomy prolapse. We have reported a simple local repair with reconstruction of the loop colostomy. We herein report a new more simple technique to avoid laparotomy and allow excision of the irreducible colostomy prolapse and complete closure of the distal limb of loop colostomy when no decompression is required in the distal limb of the stoma. In this procedure, the number of stapler and the time with blood loss for the operation can be saved.
Assuntos
Doenças do Colo/cirurgia , Colostomia/efeitos adversos , Grampeadores Cirúrgicos , Perda Sanguínea Cirúrgica , Humanos , Prolapso , Fatores de TempoRESUMO
BACKGROUND: Although patients with Alzheimer's disease and other cognitive-related neurodegenerative disorders may benefit from early detection, development of a reliable diagnostic test has remained elusive. The penetration of digital voice-recording technologies and multiple cognitive processes deployed when constructing spoken responses might offer an opportunity to predict cognitive status. OBJECTIVE: To determine whether cognitive status might be predicted from voice recordings of neuropsychological testing. DESIGN: Comparison of acoustic and (para)linguistic variables from low-quality automated transcriptions of neuropsychological testing (n = 200) versus variables from high-quality manual transcriptions (n = 127). We trained a logistic regression classifier to predict cognitive status, which was tested against actual diagnoses. SETTING: Observational cohort study. PARTICIPANTS: 146 participants in the Framingham Heart Study. MEASUREMENTS: Acoustic and either paralinguistic variables (e.g., speaking time) from automated transcriptions or linguistic variables (e.g., phrase complexity) from manual transcriptions. RESULTS: Models based on demographic features alone were not robust (area under the receiver-operator characteristic curve [AUROC] 0.60). Addition of clinical and standard acoustic features boosted the AUROC to 0.81. Additional inclusion of transcription-related features yielded an AUROC of 0.90. CONCLUSIONS: The use of voice-based digital biomarkers derived from automated processing methods, combined with standard patient screening, might constitute a scalable way to enable early detection of dementia.
Assuntos
Disfunção Cognitiva , Humanos , Disfunção Cognitiva/diagnóstico , Idioma , Sensibilidade e Especificidade , Biomarcadores , CogniçãoRESUMO
BACKGROUND: Robot-assisted laparoscopic surgery has several advantages over conventional laparoscopy. However, population-based comparative studies for low anterior resection are limited. This article aimed to compare peri-operative results of robot-assisted low anterior resection (RALAR) and laparoscopy. METHODS: This retrospective cohort study used data from patients treated with RALAR or conventional laparoscopic low anterior resection (CLLAR) between October 2018 and December 2019, as recorded in the Japanese National Clinical Database, a data set registering clinical information, perioperative outcomes, and mortality. Of note, the registry does not include information on the tumour location (centimetres from the anal verge) and diverting stoma creation. Perioperative outcomes, including rate of conversion to open surgery, were compared between RALAR and CLLAR groups. Confounding factors were adjusted for using propensity score matching. RESULTS: Of 21 415 patients treated during the study interval, 20 220 were reviewed. Two homogeneous groups of 2843 patients were created by propensity score matching. The conversion rate to open surgery was significantly lower in the RALAR group than in the CLLAR group (0.7 versus 2.0 per cent; P < 0.001). The RALAR group had a longer operating time (median: 352 versus 283 min; P < 0.001), less intraoperative blood loss (15 versus 20 ml; P < 0.001), a lower in-hospital mortality rate (0.1 versus 0.5 per cent; P = 0.007), and a shorter postoperative hospital stay (median: 13 versus 14 days; P < 0.001) compared with the CLLAR group. The CLLAR group had a lower rate of readmission within 30 days (2.4 versus 3.3 per cent; P = 0.045). CONCLUSION: These data highlight the reduced conversion rate, in-hospital mortality rate, intraoperative blood loss, and length of postoperative hospital stay for rectal cancer surgery in patients treated using robot-assisted laparoscopic surgery compared with laparoscopic low anterior resection.
Assuntos
Laparoscopia , Neoplasias Retais , Robótica , Humanos , Japão/epidemiologia , Neoplasias Retais/cirurgia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The role of Ser-308 of murine D-aspartate oxidase (mDASPO), particularly its side chain hydroxyl group, was investigated through the use of site-specific mutational analysis of Ser-308. Recombinant mDASPO carrying a substitution of Gly, Ala, or Tyr for Ser-308 was generated, and fused to either His (His-mDASPO), or glutathione S-transferase, His, and S (GHS-mDASPO) at its N-terminus. Wild-type His-mDASPO or GHS-mDASPO or their mutant derivatives were expressed in Escherichia coli and purified by affinity chromatography. All purified recombinant proteins had functional DASPO activity. The Gly-308 and Ala-308 mutants had significantly higher catalytic efficiency towards D-Asp and N-methyl-D-Asp, and a higher affinity for flavin adenine dinucleotide (FAD) compared to the wild-type enzyme. The Tyr-308 mutant had lower catalytic efficiency and binding capacity. These results suggest that the side chain hydroxyl group of a critical residue of mDASPO, Ser-308, down-regulates enzymatic activity, substrate binding, and FAD binding. This study provides information on the active site of DASPO that will considerably enhance our understanding of the biological significance of this enzyme.
Assuntos
Substituição de Aminoácidos , D-Aspartato Oxidase/química , Serina/química , Animais , Sítios de Ligação/genética , Catálise , D-Aspartato Oxidase/genética , D-Aspartato Oxidase/metabolismo , Flavina-Adenina Dinucleotídeo/química , Flavina-Adenina Dinucleotídeo/metabolismo , Camundongos , Mutagênese Sítio-Dirigida , Ligação Proteica/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Serina/genéticaRESUMO
A synthetic pathway was engineered in Escherichia coli to produce isopropanol by expressing various combinations of genes from Clostridium acetobutylicum ATCC 824, E. coli K-12 MG1655, Clostridium beijerinckii NRRL B593, and Thermoanaerobacter brockii HTD4. The strain with the combination of C. acetobutylicum thl (acetyl-coenzyme A [CoA] acetyltransferase), E. coli atoAD (acetoacetyl-CoA transferase), C. acetobutylicum adc (acetoacetate decarboxylase), and C. beijerinckii adh (secondary alcohol dehydrogenase) achieved the highest titer. This strain produced 81.6 mM isopropanol in shake flasks with a yield of 43.5% (mol/mol) in the production phase. To our knowledge, this work is the first to produce isopropanol in E. coli, and the titer exceeded that from the native producers.
Assuntos
2-Propanol/metabolismo , Vias Biossintéticas , Escherichia coli/genética , Escherichia coli/metabolismo , Acetil-CoA C-Acetiltransferase/genética , Oxirredutases do Álcool/genética , Proteínas de Bactérias/genética , Carboxiliases/genética , Clonagem Molecular , Clostridium acetobutylicum/enzimologia , Clostridium acetobutylicum/genética , Clostridium beijerinckii/enzimologia , Clostridium beijerinckii/genética , Escherichia coli/enzimologia , Expressão Gênica , Thermoanaerobacter/enzimologia , Thermoanaerobacter/genéticaRESUMO
The concentrations of free amino acids in plasma change coordinately and their profiles show distinctive features in various physiological conditions; however, their behavior can not always be explained by the conventional flow-based metabolic pathway network. In this study, we have revealed the interrelatedness of the plasma amino acids and inferred their network structure with threshold-test analysis and multilevel-digraph analysis methods using the plasma samples of rats which are fed diet deficient in single essential amino acid. In the inferred network, we could draw some interesting interrelations between plasma amino acids as follows: 1) Lysine is located at the top control level and has effects on almost all of the other plasma amino acids. 2) Threonine plays a role in a hub in the network, which has direct links to the most number of other amino acids. 3) Threonine and methionine are interrelated to each other and form a loop structure.
Assuntos
Aminoácidos Essenciais/deficiência , Lisina/sangue , Metionina/sangue , Treonina/sangue , Aminoácidos Essenciais/farmacologia , Animais , Dieta , Masculino , Redes e Vias Metabólicas , Ratos , Ratos Sprague-DawleyRESUMO
Creatol (CTL, 5-hydroxycreatinine) is a creatinine (Cr) oxidative metabolite, which was originally isolated from the urine of patients with chronic renal failure, representing a candidate for a uremic toxin. The effectiveness of CTL as an indicator of oxidative stress after kidney transplantation has not been reported. Therefore, we examined the relation between the change in oxidative stress (using CTL) in renal transplant patients and their change in renal function (using Cr). The serum Cr and serum and urine CTL were examined in five renal transplant patients. Serum CTL closely correlated with serum Cr. Serum CTL also correlated with urine CTL, but in some cases there was a time lag. Both the ratio of CTL/(Cr) and serum CTL observed in patient 2 were slow to improve after transplantation. The process through which oxidative stress was reduced was shown by the index of renal damage correlated with kidney function oxidative stress (using serum CTL) after transplantation. Our data suggested that CTL/Cr may be a good indicator of graft prognosis.
Assuntos
Creatinina/análogos & derivados , Creatinina/sangue , Testes de Função Renal , Transplante de Rim/fisiologia , Sistema ABO de Grupos Sanguíneos , Biomarcadores/sangue , Incompatibilidade de Grupos Sanguíneos , Creatinina/urina , Família , Feminino , Humanos , Falência Renal Crônica/sangue , Falência Renal Crônica/urina , Doadores VivosRESUMO
8-Hydroxy-2'-deoxyguanosine (8-OHdG) is an oxidant of deoxyguanosine, a base in the construction of DNA. We examined the extent of DNA damage to the graft through oxidative stress after renal transplantation. Seven renal transplant patients were enrolled in this study. Before reperfusion of the grafts, the level of serum 8-OHdG was measured using enzyme-linked immunosorbent assay. No relationship was found between the level of serum 8-OHdG just before reperfusion and the postoperative course. In all cases, the level of serum 8-OHdG increased after reperfusion and decreased within 2 hours. In six cases, it remained almost the same as the preoperative level. A faster rate of decrease from the first peak of serum 8-OHdG was associated with a lower nadir serum creatinine and reduced occurrence of acute rejection. This study suggested that immediate calming of the oxidative stress following reperfusion may potentially have a positive influence on graft prognosis. In addition, biomarkers of oxidative stress may predict graft prognosis.
Assuntos
Desoxiguanosina/análogos & derivados , Transplante de Rim/fisiologia , Doadores Vivos , 8-Hidroxi-2'-Desoxiguanosina , Adulto , Creatinina/sangue , Desoxiguanosina/sangue , Monitoramento Ambiental/métodos , Monitoramento Epidemiológico , Feminino , Rejeição de Enxerto/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Operatório , Prognóstico , Resultado do TratamentoRESUMO
A re-evaluation of the dielectric studies on isolated mitochondria (Pauly, H., Packer, L. and Schwan, H.P. (1960) J. Biophys. Biochem. Cytol. 7, 589-601, and ibid. 7, 603-612) is presented. The suspensions of 'mitoplasts' prepared from rat liver mitochondria by a hyposmotic (10 mM KCl) treatment showed a dielectric dispersion with its characteristic frequency lying in the 1-100 MHz range. In the analysis of data special emphasis was put on the choice of the theoretical models to employ after scrutiny of their applicability to the suspensions tested. As such we adopted the theory of Hanai et al. (Hanai, T., Asami, K., and Koizumi, N. (1979) Bull. Inst. Chem. Res., Kyoto Univ. 57, 297-305) that was advanced to include concentrated suspensions of shelled spheres. Curve fittings based on that theory resulted in a better agreement with experiment than the fittings based on a conventional theory for dilute suspensions. Major findings from our analyses on the swollen mitoplasts are that: (i) the limiting membrane of the mitoplasts has a specific electrical capacity of 1 microF/cm2, (ii) the ratio of permittivity (or dielectric constant) for the mitoplast interior and permittivity for the external medium is 0.6-0.7, and (iii) the conductivity ratio between the interior phase and the medium is approx. 0.6. Reasons for discrepancy between the results of Pauly et al. and ours are discussed.
Assuntos
Mitocôndrias Hepáticas/fisiologia , Modelos Biológicos , Animais , Eletrodos , Eletrofisiologia , Matemática , Microscopia Eletrônica , RatosAssuntos
Deficiência Intelectual/diagnóstico , Espasmos Infantis/diagnóstico , Microglobulina beta-2/urina , Doença Aguda , Adolescente , Biomarcadores/urina , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Deficiência Intelectual/urina , Japão , Síndrome de Lennox-Gastaut , Masculino , Espasmos Infantis/urinaRESUMO
We report what we believe to be the first known case of linear nevus sebaceus syndrome with infantile spasms followed by Lennox-Gastaut syndrome. The EEG revealed hemihypsarrhythmia and then asymmetrical slow spike-wave complexes. A pneumoencephalogram showed dilation and distortion of the lateral ventricle ipsilateral to the facial nevus on the left side. A computerized tomographic scan disclosed low density in the left parietotemporal region, a feature not reported previously. The findings indicated extensive lateralized cerebral lesions.
Assuntos
Nevo Pigmentado/complicações , Neoplasias Cutâneas/complicações , Espasmos Infantis/complicações , Biópsia , Encéfalo/diagnóstico por imagem , Ventriculografia Cerebral , Eletroencefalografia , Humanos , Lactente , Deficiência Intelectual/complicações , Masculino , Pneumoencefalografia , Pele/patologia , Síndrome , Tomografia Computadorizada por Raios XRESUMO
We report compositional changes in glycoconjugates in mouse kidney cortex due to aging, as analyzed by lectin histochemistry and Western blot analysis. Mouse kidney tissues of prenatal and postnatal ages (prenatal, 19 days of gestation; postnatal 2 and 8 days, 4 and 13 weeks, and 10 months) were fixed in 4% paraformaldehyde and cryosections were made. They were stained with 16 kinds of biotinylated lectin, followed by ABC, for light microscopy. Tissue homogenate was also examined by Western blotting for WGA, ConA, and Lotus. Changes in glycoconjugates due to prenatal and postnatal aging were detected by both lectin histochemistry and Western blotting.
Assuntos
Envelhecimento/metabolismo , Glicoconjugados/metabolismo , Rim/metabolismo , Lectinas , Animais , Western Blotting , Sequência de Carboidratos , Técnicas Imunoenzimáticas , Camundongos , Dados de Sequência MolecularRESUMO
We report on the immunohistochemical demonstration of an enzyme at the electron microscopic level using specimens processed by rapid freezing and the freeze-substitution technique without the use of any chemical fixatives. Fresh rat liver tissue blocks were rapidly frozen by the metal contact method using liquid nitrogen, and were freeze-substituted with acetone without any chemical fixatives at -80 degrees C. Some of the freeze-substituted tissues were embedded in Lowicryl K4M at -20 degrees C; the others were returned to room temperature and embedded in Epok 812 at 60 degrees C. Ultra-thin sections were stained using anti-peroxisomal catalase antibody by the protein A-gold technique. The ultrastructure of the hepatocytes was very well preserved compared with that of conventionally processed tissues. The labeling for catalase was confined to peroxisomes. When the labeling density was compared among freeze-substituted tissues and conventionally processed tissues, that of freeze-substituted and Lowicryl K4M-embedded tissues was the most intense. These results show the usefulness of freeze-substituted tissues for immunohistochemical analysis of cell organelles.
Assuntos
Catalase/metabolismo , Congelamento , Técnicas Histológicas , Imuno-Histoquímica/métodos , Fígado/enzimologia , Animais , Fixadores , Ouro , Fígado/citologia , Fígado/ultraestrutura , Microcorpos/ultraestrutura , Microscopia Eletrônica , Ratos , Resinas Vegetais/farmacologia , Proteína Estafilocócica ARESUMO
We studied the localization of calcineurin by immunoblotting analysis and immunohistochemistry as a first step in clarifying the role of calcineurin in the retina. Rat, bovine, and human retinal tissues were examined with subtype-nonspecific and subtype-specific antibodies for the A alpha and A beta isoforms of its catalytic subunit. In mature retinas of the three species, calcineurin was localized mainly in the cell bodies of ganglion cells and the cells in the inner nuclear layer, in which amacrine cells were distinctively positive. The calcineurin A alpha and A beta isoforms were differentially localized in the nucleus and the cytoplasm of the ganglion cell, respectively. Calcineurin was also present in developing rat retinas, in which the ganglion cells were consistently positive for it. The presence of calcineurin across mammalian species and regardless of age shown in the present study may reflect its importance in visual function and retinal development, although its function in the retina has not yet been clarified. (J Histochem Cytochem 49:187-195, 2001)
Assuntos
Calcineurina/metabolismo , Retina/crescimento & desenvolvimento , Retina/metabolismo , Idoso , Idoso de 80 Anos ou mais , Animais , Animais Recém-Nascidos , Bovinos , Humanos , Immunoblotting , Imuno-Histoquímica , Masculino , Ratos , Ratos Wistar , Retina/embriologiaRESUMO
We examined the immunohistochemical distribution of the two mammalian isoforms of calcineurin catalyic subunits, A alpha and A beta, in central nervous system (CNS) tissues of cows, rats, and humans. Cryostat sections and paraffin sections of parformaldehyde-fixed tissues were stained with antipeptide antibodies for each isoform. The same localization pattern was observed in both cryostat and paraffin sections. In the latter, the intensity of the staining was dramatically enhanced by microwave irradiation. Calcineurin isoforms were localized in a variety of nerve cells but not in neuroglial cells. Their differential expression as the A alpha isoform in the nucleus and the A beta isoform in the cytoplasm was present in a variety of CNS nerve cells, most distinctively in Purkinje cells of the cerebellum and pyramidal cells of the cerebrum, irrespective of species. These results suggest that each isform has distinct intracellular sites of action in CNS neurons and that the phenomenon has been conserved during mammalian evolution.
Assuntos
Proteínas de Ligação a Calmodulina/análise , Sistema Nervoso Central/química , Micro-Ondas , Fosfoproteínas Fosfatases/análise , Sequência de Aminoácidos , Animais , Antígenos/análise , Calcineurina , Formaldeído , Humanos , Imuno-Histoquímica , Masculino , Dados de Sequência Molecular , Inclusão em Parafina , Ratos , Ratos Wistar , Fixação de TecidosRESUMO
PURPOSE: The extracellular materials (ECMs) in the trabecular meshwork (TM) are thought to play a crucial role in aqueous outflow resistance. Immunohistochemical localization of elastin, one of the major ECMs in the normal and glaucomatous human TM, was examined ultrastructurally. METHODS: Eight normal eye bank eyes and 16 trabeculectomy specimens of primary open angle glaucoma (POAG, 11 eyes from 8 cases), congenital glaucoma (2 eyes from 1 case), and juvenile glaucoma (3 eyes from 2 cases) were embedded in Lowicryl K4M at low temperature. The distribution of elastin was studied by the protein A-gold technique. RESULTS: In normals, the gold particles indicating the antigenic sites for elastin existed mainly in the central amorphous element of the elastic-like fibers, and a few gold particles were observed within the area containing fine granular-like material and fine fibrillar-like material. No labeling was observed in cellular materials or other ECMs. In congenital and juvenile glaucoma, labeling was similar to that observed in normals. In POAG specimens compared to normals, there was an increased amount of elastin-bound immunogold particles along the inner canal endothelium. The increased gold particles, which did not have a fibrillar arrangement and were not enclosed by electron-dense microfibrils, were found within the area containing fine fibrillar-like material. However, labeling within the elastic-like fibers was similar to that observed in normals. CONCLUSIONS: Under electron microscopy, elastin could be localized in the normal and glaucomatous human TM. The results of this investigation suggest that elastin may play an important role in the etiology of POAG.
Assuntos
Elastina/metabolismo , Glaucoma de Ângulo Aberto/metabolismo , Glaucoma/metabolismo , Malha Trabecular/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias , Pré-Escolar , Glaucoma/congênito , Glaucoma/cirurgia , Glaucoma de Ângulo Aberto/cirurgia , Coloide de Ouro , Humanos , Imuno-Histoquímica , Microscopia Imunoeletrônica , Pessoa de Meia-Idade , TrabeculectomiaRESUMO
New polymeric solid-phase matrices for cell affinity chromatography were prepared and their advantageous characteristics compared with conventional matrices were highlighted. These new matrices are derivatives of poly(2-hydroxyethyl methacrylate) (PHEMA) containing a slight quantity of amino compounds as a co-monomer. They were applied to immunoaffinity selection between IgG+ and IgG- lymphocytes of the rat mesenteric lymph node. Simple physical adsorption was sufficient for anti-rat IgG antibodies to be immobilized on these matrices, allowing us to omit the laborious procedure of covalent-linking of antibodies on a matrix. As these matrices themselves showed extremely low non-specific adsorption of lymphocytes, a very dilute solution of antibody (0.02-0.08 mg/ml) was enough for column conditioning. This separation method gave IgG- lymphocytes of more than 90% purity and almost 95% yield within as short a time as 7 min. Further, IgG+ lymphocytes were obtained in good yield (80-90% of loaded number) by recovering the adsorbing cell fraction from the column by gentle pipetting of the matrix.