RESUMO
Microorganisms residing within a biofilm become more tolerant to antibiotics and other types of adverse impact, and biofilm formation by pathogenic bacteria is an important problem of current medicine. Polysaccharides that prevent biofilm formation are among the promising candidates to help tackle this problem. Earlier we demonstrated the ability of a potato polysaccharide galactan to inhibit biofilm formation by a Pseudomonas aeruginosa clinical isolate. Here we investigate the effect of potato galactan on P. aeruginosa biofilms in more detail. Microscopic analysis indicated that the galactan did not interfere with the adhesion of bacterial cells to the substrate but prevented the build-up of bacterial biomass. Moreover, the galactan not only inhibited biofilm formation, but partially destroyed pre-formed biofilms. Presumably, this activity of the galactan was due to the excessive aggregation of bacterial cells, which prohibited the formation and maintenance of proper biofilm architecture, or due to some other mechanisms of biofilm structure remodeling. This led to an unexpected effect, i.e., P. aeruginosa biofilms treated with an antibiotic and the galactan retained more viable bacterial cells compared to biofilms treated with the antibiotic alone. Galactan is the first polysaccharide demonstrated to exert such effect on bacterial biofilms.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Galactanos/farmacologia , Pseudomonas aeruginosa/fisiologia , Aderência Bacteriana/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microscopia de Fluorescência , Pseudomonas aeruginosa/crescimento & desenvolvimento , Solanum tuberosum/metabolismoRESUMO
The aim of this work was to compare biological activities of three variants of bacterially expressed human recombinant erythropoietin (EPO) with additional protein domains: 6His-s-tag-EPO protein carrying the s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) at the N-terminus and HBD-EPO and EPO-HBD proteins containing heparin-binding protein domains (HBD) of the bone morphogenetic protein 2 from Danio rerio at the N- and C-termini, respectively. The commercial preparation Epostim (LLC Pharmapark, Russia) produced by synthesis in Chinese hamster ovary cells was used for comparison. The EPO variant with the C-terminal HBD domain connected by a rigid linker (EPO-HBD) possesses the best properties as compared to HBD-EPO with the reverse domain arrangement. It was ~13 times more active in vitro (i.e., promoted proliferation of human erythroleukemia TF-1 cells) and demonstrated a higher rate of association with the erythropoietin receptor. EPO-HBD also exhibited the greatest binding to the demineralized bone matrix (DBM) and more prolonged release from the DBM among the four proteins studied. Subcutaneous administration of EPO-HBD immobilized on DBM resulted in significantly more pronounced vascularization of surrounding tissues in comparison with the other proteins and DBM alone. Therefore, EPO-HBD displayed better performance with regard to all the investigated parameters than other examined EPO variants, and it seems promising to study the possibility of its medical use.
Assuntos
Eritropoetina/genética , Escherichia coli/genética , Domínios Proteicos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Animais , Matriz Óssea/metabolismo , Proteína Morfogenética Óssea 2/genética , Proliferação de Células/efeitos dos fármacos , Eritropoetina/biossíntese , Escherichia coli/metabolismo , Humanos , Neovascularização Fisiológica/efeitos dos fármacos , Ligação Proteica , Proteínas Recombinantes/biossíntese , Peixe-ZebraRESUMO
Staphylococcus simulans lysostaphin is an endopeptidase lysing staphylococcus cell walls by cleaving pentaglycine cross-bridges in their peptidoglycan. A synthetic gene encoding S. simulans lysostaphin was cloned in Escherichia coli cells, and producer strains were designed. The level of produced biologically active lysostaphin comprised 6-30% of total E. coli cell protein (depending on E. coli M15 or BL21 producer) under batch cultivation conditions. New methods were developed for purification of lysostaphin without affinity domains and for testing its enzymatic activity. As judged by PAGE, the purified recombinant lysostaphin is of >97% purity. The produced lysostaphin lysed cells of Staphylococcus aureus and Staphylococcus haemolyticus clinical isolates. In vitro activity and general biochemical properties of purified recombinant lysostaphin produced by M15 or BL21 E. coli strains were identical to those of recombinant lysostaphin supplied by Sigma-Aldrich (USA) and used as reference in other known studies. The prepared recombinant lysostaphin represents a potential product for development of enzymatic preparation for medicine and veterinary due to the simple purification scheme enabling production of the enzyme of high purity and antistaphylococcal activity.
Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Lisostafina/farmacologia , Staphylococcus/genética , Antibacterianos/biossíntese , Antibacterianos/isolamento & purificação , Proteínas de Bactérias/metabolismo , Biomassa , Clonagem Molecular , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Escherichia coli/metabolismo , Lisostafina/biossíntese , Lisostafina/isolamento & purificação , Peptidoglicano/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Staphylococcus/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus haemolyticus/efeitos dos fármacos , TemperaturaRESUMO
BACKGROUND: Optimization of myocardial protection during cardiac surgery with a long period of anoxia infarction using sevoflurane postconditioning of myocardium. THE AIM: to develop the optimal pharmacological postconditioning protocol with sevoflurane for infarction patients ,undergoing cardiac surgery. MATERIALS AND METHODS: Two groups were formedfor this study: CON] 00 (n-32) with aortic cross-clamping time 114±15 min and SEV100 group (n-34), where the myocardium anoxia was 119±22 minutes. According to previously developed in the pilot study Protocol, we added sevofturane in the circuit of extracorporeal circulation in a dose of 2.0 vol. % 20 minutes before removing the clamp from the aorta and the first 20 min of reperfusion in the group SE V100. In the group CON1 00 pharnacological postconditioning wasn't conducted. To assess the adequacy of the cardioprotection against ischemic damage in operated patients, we used the following clinical and laboratory parameters: changing the level of troponin T; the concentration of lactate and glucose as a marker of severity of anaerobic metabolism; concentration of proinflammatory cytokines IL-6, IL-8, TNF-alpha in blood serum as reperfusion injury markers. Also we used the registration of central hemodynamics data: measuring the mean invasive blood pressure; central venous pressure; Cardiac output was measured by the method of transesophageal echocardiography TEEcho-CG, calculated left ventricular ejection fraction by Simpson. We evaluated the clinical course of the perioperative period: incidence ofperioperative myocardial ischemia; the need and the duration ofuse of cardiotonic drugs in the perioperative period; the incidence of reperfusion arrhythmias; the frequency of self-recovery heart rate. RESULTS: According to the results of anaerobic metabolism markers, we can conclude that the period of the myocardium anoxia ofpatients in both groups experienced no significant difference. However; a completely different pattern was observed when comparing the proinflammatory cytokines, such as IL-6, IL-8, TNF-a. This confirms that the group SEV] 00 survived the reperfusion is much better than the group CON100. Instrumental examination also showed that the group ofpatients in which pharmacological postconditioning with sevofturane was held signficantly better suffered ischemia and reperfiision injury compared to control group. Self-recovery heart rate after removing the aorta clamp in the group CON100 was observed in 81%, in group SEV100 same - 93%. Similarly, the frequency of myocardial ischemia episodes on the ECG in reperfusion period was two times lower in the group SEV100 compared with group CON100 - 5.8% and 12.5% respectively. Reperfusion arrhythmia is almost 3 times more frequent in the group CON100 - 21,8%, in the group SEV100, where he conducted pharmacological postconditioning with sevoflurane is 8.8%.. CONCLUSION: Combined with sevoflurane cardioprotection FPC has a much better resistance to myocardial ischemia-reperfusion injury in patients with myocardial infarction time over 100 minutes than monoprotection with cardioplegic solution "Console ". This method can be recommended as an additional method ofprotection against myocardial ischemia-reperfusion injury.
Assuntos
Procedimentos Cirúrgicos Cardíacos , Pós-Condicionamento Isquêmico/métodos , Éteres Metílicos/uso terapêutico , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Reperfusão Miocárdica/métodos , Biomarcadores/metabolismo , Humanos , Éteres Metílicos/administração & dosagem , Traumatismo por Reperfusão Miocárdica/metabolismo , Sevoflurano , Resultado do TratamentoRESUMO
Pseudomonas aeruginosa is one of the most widespread and troublesome opportunistic pathogens that is capable of colonizing various human tissues and organs and is often resistant to many currently used antibiotics. This resistance is caused by different factors, including the acquisition of specific resistance genes, intrinsic capability to diminish antibiotic penetration into the bacterial cell, and the ability to form biofilms. This situation has prompted the development of novel compounds differing in their mechanism of action from traditional antibiotics that suppress the growth of microorganisms or directly kill bacteria. Instead, these new compounds should decrease the pathogens' ability to colonize and damage human tissues by inhibiting the virulence factors and biofilm formation. The lectins LecA and LecB that bind galactose and fucose, as well as oligo- and polysaccharides containing these sugars, are among the most thoroughly-studied targets for such novel antibacterials. In this review, we summarize the results of experiments highlighting the importance of these proteins for P. aeruginosa pathogenicity and provide information on existing lectins inhibitors and their effectiveness in various experimental models. Particular attention is paid to the effects of lectins inhibition in animal models of infection and in clinical practice. We argue that lectins inhibition is a perspective approach to combating P. aeruginosa. However, despite the existence of highly effective in vitro inhibitors, further experiments are required in order to advance these inhibitors into pre-clinical studies.
RESUMO
AIM: To estimate different approaches to treatment of victims with pancreatic trauma with pancreatic trauma. MATERIAL AND METHODS: It was analyzedthe results of treatment of 342 victims with pancreatic trauma in N.V. Sklifosovskiy Research Institute of Emergency Care for the period 1991-2012. RESULTS: It was concluded that for the las decade curative and diagnostic tactics for pancreatic injury in victims with combined abdominal trauma has been changed; current diagnostic markers of pancreatic lesion and adequate intraoperative diagnosis are used. All of this together with timely specific therapy and adherence to guidelines of surgical treatment decreased mortality rate from 17.0 to 11.1% and suppurative complications incidence from 43.8 to 19.9%.
RESUMO
The sample included 43 patients with severe acute pancreatitis. Two coefficients of endogenic intoxication were applied: Kei1 = (AMP/ECA)x 100, AMP -average molecular peptides, ECA--effective concentration of albumin and Kei2 = (Kplp/aos/ECA) x 100, PLP--products of lipid peroxidation, AOS--indicators of antioxidant system. ECA--effective concentration of albumin. The comparative characteristic of both coefficients is given. It is established that Kei2 provides more informative indicators of endogenic intoxication in patients died at third day after operation. The study proved that both Kei can be applied for evaluation of endogenic intoxication and prognosis of generalized pancreonecrosis depending on resources of laboratory service.
Assuntos
Pancreatite Necrosante Aguda/sangue , Albuminas/metabolismo , Dipeptídeos/metabolismo , Humanos , Peroxidação de Lipídeos , Pancreatite Necrosante Aguda/diagnóstico , Pancreatite Necrosante Aguda/terapia , Resultado do TratamentoRESUMO
We estimated the optimal dosage of inhalation anesthetic sevoflurane, for the maximum cardioprotective effect with minimal angioparalytic action. 25 pigs were included in this study, they were divided into 5 groups, depending on the sevoflurane dosage used for pharmacological postconditioning (PPC): control group - PPC has't been conducted, a group of PPC 0.5 - sevoflurane PPC in a dose of 0.5 V%, a group of PPC 1.0 - sevoflurane PPC in a dose of 1.0 V%, a group of PPC 1.5 - sevoflurane PPC in a dose of 1.5 V%, a group of PPC 2.0 - sevoflurane PPC in a dose of 2.0 V%, a group of PPC 2.5 - sevoflurane PPC in a dose of 2.5 V%. Ischemia was simulated by left coronary artery crossclamping. Further PPC was held according to the following Protocol: 20 min before left coronary artery clamp off and first 20 min of reperfusion sevoflurane was given into CPB circuit. Myocardial ischemia period was 60 min in all groups. It was found and experimentally proved that the optimal sevoflurane dosage for PPC is 2 V%
Assuntos
Pós-Condicionamento Isquêmico/métodos , Éteres Metílicos/administração & dosagem , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Animais , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ventrículos do Coração/metabolismo , Ventrículos do Coração/patologia , Éteres Metílicos/uso terapêutico , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Sevoflurano , SuínosRESUMO
Ability of site-specific nickase BspD6I (Nt.BspD6I) to oligomerize at concentrations > or = 0.5 microM (> or = 0.035 mg/mL) is studied. Three states of Nt.BspD6I are registered via electrophoretic studies both in the presence and in the absence of DNA. Estimation of their molecular mass allows assigning them as a monomer, a dimer and a trimer. Both dimeric and monomeric Nt.BspD6I are shown to hydrolyze its DNA substrate with the identical specificity. Calculation of the electrostatic potential distribution on the Nt.BspD6I globule surface shows that the protein molecule is a dipole. The Nt. BspD6I oligomeric forms are likely to be the result of ionic protein interactions.
Assuntos
Proteínas de Ligação a DNA/química , Desoxirribonuclease I/química , Estrutura Terciária de Proteína , Bacillus/enzimologia , DNA/química , Multimerização ProteicaRESUMO
Among identified in the army for one year of active tuberculosis patients constitute 38.7% of the number of soldiers from young recruits, including 59.7% of infiltrative pulmonary tuberculosis. At the same time during the examination of these individuals revealed only about half of patients. Timely delivery of fluorographic examination recruitment plays a crucial role in preventing the spread of tuberculosis in the Armed Forces. From the military doctors need more work in groups at increased risk of tuberculosis, early identification of need in the dispensary dynamic observation and conducting a full range of therapeutic and preventive measures in foci of tuberculous infection.
Assuntos
Militares , Tuberculose Pulmonar/epidemiologia , Humanos , Radiografia Pulmonar de Massa , Militares/estatística & dados numéricos , Morbidade , Federação Russa/epidemiologia , Tuberculose Pulmonar/diagnóstico por imagem , Tuberculose Pulmonar/prevenção & controleRESUMO
Persistence is a form of interaction of pathogenic bacteria with a host aimed to promote their long-term survivalby means of inactivation of the host's protective systems via modulation of intracellular signal pathways. Persistent forms of a pathogen are refractory to traditional antibiotic therapy and cause chronic infectious diseases. Directed search for protein targets and new antibacterial drugs using computer simulation and experimental testing for the development of innovative preparations to treat chronic bacterial infections appears to have good prospects as a method for the management of persistent infections. A stepwise strategy for realization of such approach is exemplified by the search of preparations against chlamydial infection.
Assuntos
Antibacterianos/farmacologia , Fenômenos Fisiológicos Bacterianos/efeitos dos fármacos , Infecções por Chlamydia/tratamento farmacológico , Chlamydia , Desenho de Fármacos , Animais , Proteínas de Bactérias/metabolismo , Chlamydia/efeitos dos fármacos , Chlamydia/metabolismo , Chlamydia/patogenicidade , Infecções por Chlamydia/microbiologia , Doença Crônica , Simulação por Computador , Humanos , Fatores de Virulência/metabolismoRESUMO
Toll-like receptor (TLR) signaling in naive enterocytes is rapidly inhibited, leading to the establishment of tolerance. To gain insight into tolerance at the level of the proinflammatory mitogen-activated protein kinase (MAPK) p38, we characterized TLR-mediated induction of the p38-specific phosphatase MKP-1. In cultured enterocytes, ligands of TLR3, TLR4, TLR5, and TLR9, but not TLR2, induce MKP-1 at 30-60 min, coincident with dephosphorylation of p38 following the peak of TLR ligand-induced phosphorylation. Induction of MKP-1 is blocked by inhibitors of nuclear factor (NF)-kappaB, but not of MAPK. Small interfering RNA knockdown of IkBalpha prolongs the expression of MKP-1. Rat MKP-1 promoter contains two NF-kappaB-binding sites, mutations in which additively impair lipopolysaccharide-induced transcription from the MKP-1 promoter. In the intestine, MKP-1 is expressed in the crypts, the epithelial compartment that also displays bacteria-dependent activating phosphorylation of p38. Thus, NF-kappaB-dependent expression of MKP-1 may contribute, by desensitization of p38, to the rapid establishment of unresponsiveness to several TLR ligands in enterocytes.
Assuntos
Fosfatase 1 de Especificidade Dupla/metabolismo , Enterócitos/metabolismo , NF-kappa B/metabolismo , Receptores Toll-Like/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Células CACO-2 , Fosfatase 1 de Especificidade Dupla/genética , Fosfatase 1 de Especificidade Dupla/imunologia , Enterócitos/imunologia , Enterócitos/patologia , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/genética , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Tolerância Imunológica , Imunidade Inata , Intestinos/patologia , NF-kappa B/antagonistas & inibidores , Peptídeos/farmacologia , Regiões Promotoras Genéticas/genética , Ligação Proteica/efeitos dos fármacos , RNA Interferente Pequeno/genética , Ratos , Receptores Toll-Like/genética , Receptores Toll-Like/imunologiaRESUMO
ETS proteins comprise a family of widespread transcription factors regulating expression of many animal genes. Structurally ETS proteins are characterized by conserved DNA-binding ETS domain, recognizing DNA sequences with trinucleotide GGA. Comparative analysis of structural features of ETS domain-DNA complexes was carried out and conserved contacts, important in terms of interaction stability and specificity were identified. The analysis revealed 9 conserved hydrogen bonds with DNA backbone phosphates and 2 conserved bidentate hydrogen bonds with DNA major groove atoms, one conserved hydrophobic cluster on protein-DNA interface, important for binding site recognition, and 12 conserved water molecules, possibly mediating ETS domain-DNA interaction. The results are represented in specialized data bank of protein-DNA complexes NPIDB.
Assuntos
DNA/química , Proteínas Proto-Oncogênicas c-ets/química , Animais , DNA/genética , DNA/metabolismo , Humanos , Camundongos , Ligação Proteica/fisiologia , Estrutura Terciária de Proteína/fisiologia , Proteínas Proto-Oncogênicas c-ets/genética , Proteínas Proto-Oncogênicas c-ets/metabolismoRESUMO
The use of videolaparoscopic methods for the treatment of penetrating stomach and duodenal ulcers, acute cholecystitis, acute pancreatitis, acute appendicitis, intestinal obstruction, acute gynecological diseases and abdominal trauma is analyzed. Laparoscopic methods at urgent abdominal surgery improves the quality of diagnosis and treatment, decrease the rate of postoperative complications and lethality, reduce the hospital stay.
Assuntos
Abdome/cirurgia , Laparoscopia , Cirurgia Vídeoassistida , Traumatismos Abdominais/cirurgia , Adulto , Algoritmos , Apendicectomia/métodos , Apendicite/cirurgia , Colecistite Aguda/cirurgia , Úlcera Duodenal/cirurgia , Emergências , Feminino , Doenças dos Genitais Femininos/cirurgia , Humanos , Obstrução Intestinal/cirurgia , Laparoscopia/mortalidade , Laparotomia , Tempo de Internação , Masculino , Pancreatite/cirurgia , Gravidez , Complicações na Gravidez/cirurgia , Úlcera Gástrica/cirurgia , Fatores de TempoRESUMO
Results of examination and treatment of 958 patients with acute pancreatitis are analyzed. The study group consisted of 372 patients treated in accordance with special clinical protocols (2001-2004). The control group consisted of 586 patients treated before these protocols extensive use (1995-2001). The two groups were similar by the main clinical characteristics. It was demonstrated that the treatment according standard protocols permitted to reduce general lethality from 3.9% (control group) to 2.7% (study group), postoperative lethality - from 14.3 to 11.5%. General lethality at severe acute pancreatitis decreased from 18.5 to 12.7%, postoperative lethality - from 26 to 19%.
Assuntos
Antibacterianos/uso terapêutico , Imunossupressores/uso terapêutico , Laparoscopia/métodos , Octreotida/uso terapêutico , Pancreatite Necrosante Aguda/diagnóstico , Pancreatite Necrosante Aguda/terapia , Esfinterotomia Endoscópica/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia por Agulha Fina , Diagnóstico Diferencial , Feminino , Seguimentos , Fármacos Gastrointestinais/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Pancreatite Necrosante Aguda/mortalidade , Estudos Retrospectivos , Federação Russa/epidemiologia , Taxa de Sobrevida , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
Sarcoidosis is a chronic multi-system granulomatous disease of unknown etiology, which is characterized by development of epitheliocellular granulomas. Its therapy is not substantiated enough. The incidence of sarcoidosis among the called up servicemen is 1,1 and among the contract servicemen--3,5. According to the structure of contingents with initial diagnosis the called up servicemen constitute 18,6%, officers and praporschiks--57,7% and the contract servicemen without commissioned ranks--23,7%. The comparative analysis has shown that among the contract servicemen suffering from sarcoidosis the number of women was 2,1 times more than among the patients with tuberculosis. The medical personnel in the sarcoidosis group is observed more often (1,5 times more) than in the tuberculosis.
Assuntos
Assistência Ambulatorial/métodos , Militares , Sarcoidose Pulmonar , Adulto , Diagnóstico Diferencial , Feminino , Humanos , Incidência , Masculino , Federação Russa , Sarcoidose Pulmonar/complicações , Sarcoidose Pulmonar/diagnóstico , Sarcoidose Pulmonar/epidemiologia , Sarcoidose Pulmonar/terapia , Tuberculose dos Linfonodos/complicações , Tuberculose dos Linfonodos/diagnóstico , Tuberculose dos Linfonodos/epidemiologiaRESUMO
Results of different x-ray methods applied in 151 patients early after operations for destructive pancreatitis were analyzed. The technique of fistulography is described. Plain roentgenography of the chest demonstrated changes in 77.4% cases, most often occurred disk-shaped atelectases and pleural effusion. Examination of the abdominal cavity revealed functional disorders in 69.8% cases, direct symptoms of purulent process -- in 52.9%, radiographic signs of aseptic sequestration -- in 47.1% patients. X-ray signs of purulent process in the retroperitoneal fat and aseptic sequestration of the infiltrate or fat are outlined. The value of fistulography for localization and determination of the shape of drained cavities, position of the drainage tubes in the cavity, condition of the adjacent tissue, progression of purulent process, adequacy of drain of the purulent cavities is shown. Based on the results of complex x-ray examination with ultrasound investigation and CT data, the indications to repeated procedure (surgery or additional drainage) were formulated in 82 (54.3%) of 151 patients.
Assuntos
Fístula/diagnóstico por imagem , Fístula/etiologia , Omento/diagnóstico por imagem , Pancreatite Necrosante Aguda/diagnóstico por imagem , Pancreatite Necrosante Aguda/cirurgia , Doenças Peritoneais/diagnóstico por imagem , Doenças Peritoneais/etiologia , Complicações Pós-Operatórias , Tomografia Computadorizada por Raios X , Fístula/epidemiologia , Humanos , Doenças Peritoneais/epidemiologiaRESUMO
The results of the immunological study of blood in 89 patients with severe acute pancreatitis (SAP) were analyzed. Immune reactions in SAP were characterized by the presence of moderate leukocytosis, an increase in the phagocytic and metabolic activity of neutrophils, relative lymphopenia, a decrease in the number of B-lymphocytes. The average values of the parameters of the immunogram of patients with the favorable course of the disease on days 2 - 3 were analogous to the average population values in 89 examinees. The direction and the degree of manifestation were slightly variable and could be registered within one standard quadratic deviation. Such heterogeneity in the character of immune reactions made it possible to regard the average values of the immunogram parameters of these patients as "the norm". For the objective evaluation of the immune status of SAP patients a scale for the evaluation, in points, of disturbances in immune reactions to the destructive process in the pancreas was developed. This scale for the evaluation, in points, of the severity of disturbances in the immune status of SAP patients made it possible to prognosticate the development of complications.
Assuntos
Pancreatite Necrosante Aguda/imunologia , Adulto , Linfócitos B/imunologia , Feminino , Humanos , Leucocitose/imunologia , Linfopenia/imunologia , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Neutrófilos/metabolismo , Pancreatite Necrosante Aguda/diagnóstico , Fagocitose , PrognósticoRESUMO
Some nuclear proteins of human HeLa and HepG2 cells are capable of binding to GCC-triplet repeats--(GCC)n > 3 in 5'-regulatory regions of a number of mammalian genes--G-C-elements. According to our previous data, nucleotide sequence (GCC)4 in promoter of mouse ribosomal protein L32 gene (rpL32) between 17 and 6 bp upstream of transcription start site interacts to nuclear proteins from HepG2 cells, and may be considered as a GCC-element. We suggest that one of those proteins, with molecular weight about 52 kDa, which may interact with rpL32 GCC-element, is a known conservative mammalian transcription factor ZF5. DNA-binding domain of ZF5 contains a few Kruppel-like Zn-fingers (Cys2His2-type) interacting with the GC-rich nucleotide sequences in 5'-regulatory regions of a number of mammalian genes. Our results (obtained by EMSA) showed that recombinant GST-ZF5 fused protein containing ZF5 DNA-binding domain specifically binds a few GS-rich sequences: (GCC)g-9riplet repeats, 5'-GCGCGC-3' (known ZF5 consensus binding site) and (more preferable) the fragment (-24...+1 bp) of rpL32 promoter. The high affinity of ZF5 DNA-domain binding with the latter may be explained by the presence in this fragment of two overlapped subsequences, each being capable of binding to ZF5: (GCC)4 and 5'-GCGCGC- 3'. Zf5 cDNA was cloned from HepG2 cells by RT-PCR method, and then used for construction of the gene expression vector. It has been shown that Zf5 cDNA expression vector specifically down-regulates (in luciferase assays) the activity of rpL32 promoter (-155...+159) including the above mentioned GC-rich subsequences by cotransfection of HepG2 cells. Therefore, our results enable us to consider GCC-elements as a novel class of ZF5 targets in 5'-regulatory regions of mammalian genes.
Assuntos
Proteínas de Ligação a DNA/genética , Genes Reguladores , Proteínas Recombinantes de Fusão/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Repetições de Trinucleotídeos , Linhagem Celular Tumoral , DNA Complementar , Regulação Neoplásica da Expressão Gênica , Humanos , Fatores de Transcrição Kruppel-Like , Reação em Cadeia da Polimerase , Regiões Promotoras GenéticasRESUMO
An analysis of data of immunological investigation of blood of 89 patients with a severe acute pancreatitis (SAP) on the 2-3 day of the disease has revealed changes in immunogram characteristic also of other pathological conditions and pointing to a systemic inflammatory reaction. Among the patients who did not have purulent complications against the background of therapy of SAP, the character and degree of changes in immunogram were in line with average values, and so they could be considered as "a norm of pathology". A system of scores of impairments of the immune response to the destructive process was developed for an objective evaluation of the immune status of patients with SAP. In patients with a normal physiological immune response (sum of scores 0-5) the prognosis of the disease is considered to be good. No immunocorrectors should be included in the complex of treatment measures. In patients with an inadequate pathological immune response (sum of scores 9 and more) the prognosis is unfavorable. The complex of treatment measures for such patients should include immunocorrectors.