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1.
Biomed Pharmacother ; 111: 1147-1155, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30841428

RESUMO

This article describes the work on synthesis, surface functionalization and characterization of hollow gold nanoshells (HGNs), and their application in a photothermal tumor therapy. The studied gold nanoshells were synthesized using a method based on a reduction of HAuCl4 onto silver nanoparticle templates. A selected aptamer - AS1411 - selective towards nucleolin, with a terminal thiol group was conjugated to HGNs. Surface functionalization of synthesized nanoparticles was necessary to gain the affinity to tumor cells, thus allowing the selective delivery of the modified nanoparticles to target cells. As expected, an increased selectivity of nanoconjugates towards tumor cells (A375) in a comparison to normal ones (HaCaT) was observed (2.67 times). In the framework of our studies the biological activity of the obtained nanoconjugates was evaluated using MTT assay. For this aim, both normal and tumor cell cultures respresenting skin tissue were exposed to gold nanoparticles solutions. It was found, that HGN-AS1411 conjugates cause less than 10% loss in a cell viability in the case of both cell lines at the highest tested concentration. The potential of the developed nanoconjugates as agents in PTT of skin cancer was investigated in a subsequent stage of our research. It was found, that A375 cell viability amounted to less than 40% for 75µM of nanoconjugates (expressed as a concentration of Au atoms) as a result of the laser irradiation of this cell culture after desired accumulation of the tested nanoconjugates inside A375 cells.


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Nanoconchas/química , Linhagem Celular , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Nanopartículas Metálicas/administração & dosagem , Nanoconchas/administração & dosagem , Fototerapia/métodos , Prata/química , Neoplasias Cutâneas/tratamento farmacológico , Nanomedicina Teranóstica/métodos
2.
Radiat Prot Dosimetry ; 180(1-4): 177-181, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29194515

RESUMO

Nanodosimetry is a branch of dosimetry for investigation and modeling of the interaction pattern of ionizing radiation in nanometre site-sizes (at unit density), which dates back to the 1970's (Pszona S. A track ion counter. Proceedings of Fifth Symposium on Microdosimetry EUR 5452 d-e-f, Published by the Commission of the European Communities, Luxemburg, pp. 1107-1122 (1976)). To date, the different experimental approaches have lead to developing of three fully functional nanodosimeters: the Jet Counter operated at NCBJ, the Ion Counter operated at PTB and Startrack Counter operated at INFN-LNL. Descriptions of each nanodosimeter as well as of the techniques used to investigate the track structure of ionizing particles are presented.


Assuntos
Modelos Teóricos , Nanotecnologia/instrumentação , Aceleradores de Partículas/instrumentação , Monitoramento de Radiação/instrumentação , Doses de Radiação
3.
Radiat Prot Dosimetry ; 180(1-4): 150-156, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29036364

RESUMO

The biological action of ionizing charged particles is initiated at the DNA level, and the effectiveness with which the initial physical effect changes into measurable biological damage is likely ruled by the stochastics of ionizations produced by the incident ions in subcellular nanometric volumes. Based on this hypothesis, experimental nanodosimetry aims at establishing a new concept of radiation quality that builds on measurable characteristics of the particle track structure at the nanometer scale. Three different nanodosimetric detection systems have been developed to date that allow measurements of the number of ionizations produced by the passage of a primary particle in a nanometer-size gas volume (in unit density scale). Within the Italian project MITRA (MIcrodosimetry and TRAck structure), funded by the Italian Istituto Nazionale di Fisica Nucleare (INFN) and the EMRP Joint Research Project 'BioQuaRT' (Biologically Weighted Quantities in Radiotherapy), experiments have been carried out, in which the frequency distribution of ionizations produced by proton and carbon ion beams of given energy was measured with the three nanodosimetric detectors. Descriptors of the track structure can be derived from these distributions. In particular, the first moment M1, representing the mean number of ionizations produced in the target volume, and the cumulative probability Fk of measuring a number ν ≥ k of ionizations. The correlation between measured nanodosimetric quantities and experimental radiobiological data available in the literature is here presented and discussed.


Assuntos
Nanotecnologia/métodos , Nanotecnologia/tendências , Exposição Ocupacional/análise , Garantia da Qualidade dos Cuidados de Saúde , Monitoramento de Radiação/métodos , Proteção Radiológica/métodos , Animais , Células CHO , Carbono , Sobrevivência Celular , Cricetulus , Nanotecnologia/instrumentação , Prótons , Monitoramento de Radiação/instrumentação , Proteção Radiológica/instrumentação , Radiobiologia , Medição de Risco , Gestão da Segurança
4.
J Anim Physiol Anim Nutr (Berl) ; 98(5): 914-20, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24387699

RESUMO

The effect of two diets enriched with unsaturated fatty acids--one containing the addition of dried distillers grains with solubles (DGS) and the other the addition of false flax--Camelina sativa cake (CS)--on some metabolic parameters and hormone concentration in growing lambs was determined in this experiment. A total of 21 ram lambs of the Polish Whiteheaded mutton sheep were divided into three groups (the control, receiving DGS and CS). The diets were administered to animals for 6 weeks. During the experiment, peripheral blood was collected. Glucose (GL), total cholesterol (CH), triglycerides (TG), free fatty acids (FFA), insulin (IN), leptin (LE), triiodothyronine (T3) and thyroxine (T4) were assayed in serum. The age-dependent reduction in CH and TG limited by both experimental diets were observed. A significant increase in FFA concentration was observed in samples collected in the last, that is, third, time period. This was most probably caused by a 12-h pre-slaughter fasting period. A significantly lower dynamic of FFA increase in that experimental period was found in animals receiving the experimental feed. Insulin concentration in DGS-receiving lambs was increased, in contrast to the CS-receiving lambs, in which it was lower when compared to the control. LE concentration was decreased by both experimental diets, more markedly in the DGS-receiving animals. No differences between the experimental groups and the control were observed in T3 and T4 concentrations. The effect of 12-h pre-slaughter fasting was statistically highly significant for the levels of examined blood markers and hormones, except for TG and IN in the group of lambs receiving the experimental diet with CS.


Assuntos
Ração Animal/análise , Dieta/veterinária , Gorduras Insaturadas na Dieta/administração & dosagem , Metabolismo Energético/fisiologia , Ovinos/metabolismo , Envelhecimento , Fenômenos Fisiológicos da Nutrição Animal , Animais , Gorduras Insaturadas na Dieta/metabolismo , Lipídeos/sangue , Ovinos/sangue , Ovinos/crescimento & desenvolvimento
5.
Artigo em Inglês | MEDLINE | ID: mdl-23719413

RESUMO

The UV-Visible absorption spectra of six, newly synthesized donor-substituted 2-amino-1,3-dicyano-5,6,7,8-tetrahydronaphthalene have been measured in methylcyclohexane (MCH) and assigned with the help of quantum-chemical calculations. Our calculations have been performed to assess information regarding the electronic state energy values, corresponding oscillator strengths, x-, y-, z-components of the transition dipole moments and molecular orbitals involved in the main electronic transitions of the studied compounds. Additionally, the experimental absorption transition dipole moments were calculated, on the basis of spectroscopic data, and compared with results of our quantum-chemical calculations. On the basis of the experimental results and quantum-chemical calculations, it was shown that the long-wavelength absorption band involves an overlap of three electronic transitions of different character. For all studied donor-acceptor (D-A) compounds in vapour-phase, the long-wavelength transition (S0→S1) does not possess charge transfer character, whereas the S0→S2 transition possesses electron transfer character e.g., π-electrons of the acceptor moiety are moved to the donor part. Moreover, it is found that the electronic structure of the studied biphenyl derivatives can be approximately described within composite-model of decoupled moieties: donor and acceptor.


Assuntos
Elétrons , Modelos Moleculares , Naftalenos/química , Nitrilas/química , Teoria Quântica , Absorção , Corantes/química , Conformação Molecular , Termodinâmica
6.
Biol Sport ; 30(2): 137-43, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24744480

RESUMO

The aim of the study was to assess changes in the anaerobic threshold of young soccer players in an annual training cycle. A group of highly trained 15-18 year old players of KKS Lech Poznan were tested. The tests included an annual training macrocycle, and its individual stages resulted from the time structure of the sports training. In order to assess the level of exercise capacities of the players, a field exercise test of increasing intensity was carried out on a soccer pitch. The test made it possible to determine the 4 millimolar lactate threshold (T LA 4 mmol · l(-1)) on the basis of the lactate concentration in blood [LA], to establish the threshold running speed and the threshold heart rate [HR]. The threshold running speed at the level of the 4 millimolar lactate threshold was established using the two-point form of the equation of a straight line. The obtained indicators of the threshold running speed allowed for precise establishment of effort intensity used in individual training in developing aerobic endurance. In order to test the significance of differences in mean values between four dates of tests, a non-parametric Friedman ANOVA test was used. The significance of differences between consecutive dates of tests was determined using a post-hoc Friedman ANOVA test. The tests showed significant differences in values of selected indicators determined at the anaerobic threshold in various stages of an annual training cycle of young soccer players. The most beneficial changes in terms of the threshold running speed were noted on the fourth date of tests, when the participants had the highest values of 4.01 m · s(-1) for older juniors, and 3.80 m · s(-1) for younger juniors. This may be indicative of effective application of an individualized programme of training loads and of good preparation of teams for competition in terms of players' aerobic endurance.

7.
Solid State Nucl Magn Reson ; 39(3-4): 72-80, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21316931

RESUMO

Transmission line-based multi-channel solid state NMR probes have many advantages regarding the cost of construction, number of RF-channels, and achievable RF-power levels. Nevertheless, these probes are only rarely employed in solid state-NMR-labs, mainly owing to the difficult experimental determination of the necessary RF-parameters. Here, the efficient design of multi-channel solid state MAS-NMR probes employing transmission line theory and modern techniques of electrical engineering is presented. As technical realization a five-channel ((1)H, (31)P, (13)C, (2)H and (15)N) probe for operation at 7 Tesla is described. This very cost efficient design goal is a multi port single coil transmission line probe based on the design developed by Schaefer and McKay. The electrical performance of the probe is determined by measuring of Scattering matrix parameters (S-parameters) in particular input/output ports. These parameters are compared to the calculated parameters of the design employing the S-matrix formalism. It is shown that the S-matrix formalism provides an excellent tool for examination of transmission line probes and thus the tool for a rational design of these probes. On the other hand, the resulting design provides excellent electrical performance. From a point of view of Nuclear Magnetic Resonance (NMR), calibration spectra of particular ports (channels) are of great importance. The estimation of the π/2 pulses length for all five NMR channels is presented.


Assuntos
Eletricidade , Engenharia/métodos , Espectroscopia de Ressonância Magnética/métodos
8.
Nucleosides Nucleotides Nucleic Acids ; 26(10-12): 1211-5, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-18066754

RESUMO

DcpS (scavenger decapping enzyme) from nematode C. elegans readily hydrolyzes both monomethyl- and trimethylguanosine cap analogues. The reaction was followed fluorimetrically. The marked increase of fluorescence intensity after the cleavage of pyrophosphate bond in dinucleotides was used to determine K(m) and V(max)values. Kinetic parameters were similar for both classes of substrates and only slightly dependent on pH. The hydrolysis was strongly inhibited by methylene cap analogues (m(7)Gp(CH(2))ppG and m(7)Gpp(CH(2))pG) and less potently by ARCA (m(7,3' O)GpppG).


Assuntos
Proteínas de Caenorhabditis elegans/química , Guanina/análogos & derivados , Pirofosfatases/química , Análogos de Capuz de RNA/química , Animais , Proteínas de Caenorhabditis elegans/antagonistas & inibidores , Guanina/química , Hidrólise/efeitos dos fármacos , Cinética , Pirofosfatases/antagonistas & inibidores , Análogos de Capuz de RNA/farmacologia , Espectrometria de Fluorescência
10.
Ortop Traumatol Rehabil ; 3(4): 535-7, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-17984911

RESUMO

Neurodevelopmental rehabilitation methods for children with nonprogressive brain injury began to take shape in the late 1940s in the US and Europe. One of the most important centers continuously involved in promoting neurodevelopmental concepts is the Institute for the Achievement of Human Potential. The method of child neurodevelopment is based on the theory of neurological organization. In practice this global method seeks to provide the child's brain with ordered information suitable for his or her present level of development, with increasing duration, frequency, and intensity. As is the case with other rehabilitation methods, the theoretical foundations and clinical efficacy of these rehabilitation techniques have not yet been adequately documented, which has been pointed out by the numerous critics of these methods.

11.
Neurol Neurochir Pol ; 34 Suppl 1: 67-75, 2000.
Artigo em Polonês | MEDLINE | ID: mdl-10768147

RESUMO

The paper presented the significance of EEG activation in diagnosing epilepsy in children and adolescents. EEG records of 108 patients between 3-19 years of age with the occurrence of various types of paroxysmal disorders who were admitted to Chair and Department of Developmental Neurology University of Medical Sciences in Poznan were analysed. Standard EEG was administered to all children twice: the first examination at rest with hyperventilation (HV) and photostimulation (FS) and the second one after total or partial sleep deprivation (DS). At awaking state normal records were stated in 57% of cases, in 15% paroxysmal changes in HV or FS appeared, in the rest of cases (28%) generalised paroxysmal disorders, localised changes, lateralised or diffused were noted. Among EEG activation methods DS occurred to be the most effective after it, 52 patients (48%) were diagnosed for generalised paroxysmal changes, 20% for localised, 3% lateralised and for 6% for diffused changes. Normal EEG were recorded in 22% of examined children and adolescents. The epilepsy were diagnosed in 59 (55%) of patients mostly with generalised seizures (28) partial secondarily generalised (16) and partially complex (11). In 49 (45%) children were diagnosed with other paroxysmal disorders.


Assuntos
Eletroencefalografia , Epilepsia/diagnóstico , Privação do Sono/diagnóstico , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Hiperventilação/diagnóstico , Masculino , Estimulação Luminosa/métodos , Índice de Gravidade de Doença , Vigília/fisiologia
12.
Przegl Lek ; 56(1): 48-50, 1999.
Artigo em Polonês | MEDLINE | ID: mdl-10375926

RESUMO

The objective of the paper was an evaluation of relationship between oncologically suspicious cytologic smear and colposcopic images, and the results of histopathological examination of samples collected from the vaginal portion of uterine cervix and from cervical canal scrapings. The material consisted of 60 patients with cytologic group 3 and 13 patients with group 4 and 5 according to Papanicolaou, who had been selected during a two-year preventive examination period. Cytologic group 3 patients (group 1) were subjected to cytological assessment and colposcopic examination again in order to determine further diagnostic indications. Cytologic group 4 and 5 women (group 2) were collected samples from the vaginal portion of uterine cervix under colposcopic supervision, without waiting for the result of cytologic re-assessment. All these patients were also collected samples from the vaginal portion of uterine cervix. In patients of cytologic group 3 the cytologic and colposcopic examination results were found to be compatible with histopathologic examination results in 53.3% cases. In patients of cytologic group 4 or 5 this factor was established at 92.3%.


Assuntos
Programas de Rastreamento/métodos , Neoplasias do Colo do Útero/diagnóstico , Adulto , Idoso , Colposcopia/métodos , Feminino , Humanos , Pessoa de Meia-Idade , Teste de Papanicolaou , Neoplasias do Colo do Útero/prevenção & controle , Esfregaço Vaginal
13.
Ann Emerg Med ; 32(2): 129-38, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9701293

RESUMO

STUDY OBJECTIVE: To evaluate intramuscular dihydroergotamine in direct comparison with opioid analgesia in the treatment of acute migraine headache. METHODS: This was a prospective, multicenter, double-blind trial performed in the emergency departments of 11 general hospitals in the United States. One hundred seventy-one patients between the ages of 18 and 60 years who presented to the ED with acute migraine headache were enrolled. Patients were randomly assigned to receive either 1 mg dihydroergotamine (DHE) or 1.5 mg/kg meperidine (MEP) by intramuscular injection. The anti-nauseant hydroxyzine (H) was coadministered in both treatment groups. RESULTS: One hundred fifty-six patients were evaluable. Treatment groups were comparable in sample size, demographics, and baseline measurements of headache pain. Reduction of headache pain as measured on a 100-mm visual analog scale was 41+/-33 mm (53.5% reduction) for the DHE group, and 45+/-30 mm (55.7% reduction) for the MEP group at 60 minutes after treatment (difference=2.2%; 95% confidence interval [CI] -10%, 14.5%; P=.81). Reduction in the severity of nausea and improvement in functional ability were similar between treatment groups. Central nervous system adverse events were less common in the DHE group (DHE 23.5% versus MEP 37.6%, difference-14.1%: 95% CI -28%, 0%). In particular, dizziness was reported less commonly with DHE than MEP (2% versus 15%, difference=-13%: 95% CI -21%, -5%). CONCLUSION: In this prospective, double-blind trial of a convenience sample of ED patients randomly assigned to one of two treatment regimens, DHE and MEP were comparable therapies for acute migraine. The use of DHE avoids several problems associated with opioid analgesia, including dizziness.


Assuntos
Analgésicos não Narcóticos/uso terapêutico , Analgésicos Opioides/uso terapêutico , Antieméticos/uso terapêutico , Di-Hidroergotamina/uso terapêutico , Hidroxizina/uso terapêutico , Meperidina/uso terapêutico , Transtornos de Enxaqueca/tratamento farmacológico , Doença Aguda , Adolescente , Adulto , Analgésicos não Narcóticos/administração & dosagem , Analgésicos não Narcóticos/efeitos adversos , Analgésicos Opioides/administração & dosagem , Analgésicos Opioides/efeitos adversos , Antieméticos/administração & dosagem , Antieméticos/efeitos adversos , Di-Hidroergotamina/administração & dosagem , Di-Hidroergotamina/efeitos adversos , Método Duplo-Cego , Combinação de Medicamentos , Serviço Hospitalar de Emergência , Feminino , Seguimentos , Humanos , Hidroxizina/administração & dosagem , Hidroxizina/efeitos adversos , Injeções Intramusculares , Masculino , Meperidina/administração & dosagem , Meperidina/efeitos adversos , Pessoa de Meia-Idade , Náusea/tratamento farmacológico , Medição da Dor , Estudos Prospectivos , Fatores de Tempo
14.
Arch Immunol Ther Exp (Warsz) ; 46(1): 51-7, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9510947

RESUMO

Epithelial cell cultures became an experimental model employed for both animal and human studies. We established a modified method of culture of human nasal epithelial cells from nasal polyps using serum-free, hormonally supplemented Ham's F-12 medium and Vitrogen 100 collagen matrix. Cells reached confluent monolayer structures in 5 to 7 days (mean time 6 +/- 0.89 days) of culture. The confluent cultures consisted of pure epithelial cells, which was confirmed by light and electron microscopy, and immunohistochemical staining with anticytokeratin antibody. The success ratio of cultures was 61.5%. The culture system described is efficient enough to provide pure epithelial cells for further functional studies.


Assuntos
Técnicas de Cultura de Células/métodos , Colágeno , Células Epiteliais/citologia , Pólipos Nasais , Separação Celular , Células Cultivadas , Células Epiteliais/ultraestrutura , Humanos , Imuno-Histoquímica , Queratinas
15.
J Anim Sci ; 75(8): 2106-16, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9263058

RESUMO

The pale, soft, exudative (PSE) phenomenon in turkey pectoralis major (breast) muscle was studied using a combination of biochemical, meat quality, microscopic, and gel electrophoresis techniques. Breast muscle samples were collected from turkeys characterized by slow vs fast postmortem glycolysis assessed by muscle pH at 20 min after death. The PSE group was characterized by lower muscle ATP (P < .05) and higher lactate levels (P < .05) compared with the normal group. Excess water-holding capacity and cooking yield were significantly lower (P < .05) in the PSE group than in normal turkeys. Breast muscle of the PSE group was also lighter (P < .05) than that in the normal group as determined by Minolta L* values. The SDS-PAGE, Western blotting, and immunofluorescence microscopy revealed that phosphorylase, a soluble enzyme, became tightly associated with the myofibrils in muscle from the PSE group. Also, less myosin could be solubilized from PSE vs normal myofibril samples. The results indicate that irreversible myosin insolubility due to low pH and high-temperature conditions is decisive in the development of PSE turkey breast muscle.


Assuntos
Proteínas Musculares/fisiologia , Músculos Peitorais/fisiologia , Rigor Mortis/veterinária , Perus/fisiologia , Trifosfato de Adenosina/análise , Animais , Western Blotting/métodos , Western Blotting/veterinária , Fracionamento Celular/métodos , Eletroforese em Gel de Poliacrilamida/métodos , Eletroforese em Gel de Poliacrilamida/veterinária , Concentração de Íons de Hidrogênio , Imuno-Histoquímica , Lactatos/análise , Masculino , Carne/normas , Proteínas Musculares/análise , Miofibrilas/química , Miofibrilas/fisiologia , Miofibrilas/ultraestrutura , Miosinas/análise , Miosinas/fisiologia , Músculos Peitorais/química , Fosfatos/análise , Rigor Mortis/fisiopatologia , Temperatura , Fatores de Tempo
16.
Emerg Med Clin North Am ; 15(1): 251-60, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9056579

RESUMO

There are physiologic and logistic concerns involved in caring for patients with inflight emergencies. This article presents the physiologic changes that occur at altitude, the types of medical problems that can be encountered, and management strategies. An overview is also presented of the medical training of flight attendants and the medical equipment that is available on commercial aircraft.


Assuntos
Medicina Aeroespacial , Aeronaves , Viagem , Altitude , Causas de Morte , Emergências , Pessoal de Saúde/educação , Humanos , Morbidade
17.
Ginekol Pol ; 68(12): 633-5, 1997 Dec.
Artigo em Polonês | MEDLINE | ID: mdl-9686475

RESUMO

The Potter syndrome was discussed based on the reference materials. The aim of the work is to present the course of pregnancy of 19-year-old nullipara ended by the delivery of male newborn with signs of above described syndrome.


Assuntos
Anormalidades Múltiplas/diagnóstico por imagem , Retardo do Crescimento Fetal , Rim/anormalidades , Pulmão/anormalidades , Adulto , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Resultado da Gravidez , Diagnóstico Pré-Natal , Síndrome , Ultrassonografia
19.
Am J Emerg Med ; 12(3): 326-8, 1994 May.
Artigo em Inglês | MEDLINE | ID: mdl-8179741

RESUMO

Aortic dissection usually presents with chest pain, abnormal pulses, and a widened mediastinum on chest radiograph. It is rarely associated with the superior vena cava syndrome (SVCS), which more commonly occurs in the setting of malignant disease. A patient who had SVCS as a result of a painless aortic dissection is presented and compared with other previously reported cases of simultaneous SVCS and aortic dissection.


Assuntos
Dissecção Aórtica/complicações , Síndrome da Veia Cava Superior/etiologia , Idoso , Dissecção Aórtica/fisiopatologia , Dissecção Aórtica/cirurgia , Diagnóstico Diferencial , Humanos , Masculino , Síndrome da Veia Cava Superior/diagnóstico
20.
Plant Mol Biol ; 24(5): 733-41, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8193298

RESUMO

Poly(A)+ RNA isolated from flower buds of Cynara cardunculus has been used to prepare a cDNA library. Screening of the cDNA after expression of cloned DNA with antibodies raised against the large subunit of cyprosin 3 resulted in the isolation of six positive clones. One of these clones (cypro1s; a 1.7 kb Eco RI fragment) codes for cyprosin. The nucleotide sequence contain a 1419 bp open reading frame coding for 473 amino acids (aa) including a putative full-length mature protein (440 aa) and a partial prosequence (33 aa). Cypro1s contains a 162 bp 3' non-coding region followed by a poly(A) tail. The deduced amino acid sequence shows high homology to other plant aspartic proteinases. The homology to mammalian and microbial aspartic proteinases is somewhat lower. Plant aspartic proteinases contain an insert of around 100 aa. We are modelling where this plant-specific insert will appear in the structure of cyprosin. Using cypro1s as a probe in northern blot analysis, the expression of cyprosin in developing flowers and other tissues has been studied. The signal on the northern blot increased for RNA samples from early (flower buds 6 mm in length) to later stages of floral development (flower buds up to 40 mm in length). In late stages of floral development (open flowers 50 mm in length and styles from such flowers) no hybridization signal was visualized showing that the synthesis of mRNA encoding the cyprosin starts in early stages of floral development and switches off at maturation of the flower.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Ácido Aspártico Endopeptidases/genética , Plantas/enzimologia , Plantas/genética , Sequência de Aminoácidos , Ácido Aspártico Endopeptidases/biossíntese , Quimosina/genética , Clonagem Molecular , DNA Complementar/genética , Escherichia coli/genética , Regulação da Expressão Gênica , Genoma , Modelos Moleculares , Dados de Sequência Molecular , Proteínas Recombinantes/biossíntese , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Distribuição Tecidual
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