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We present achromatic frequency doubling of supercontinuum pulses from a hollow core fiber as a technique for obtaining tunable ultrashort pulses in the near UV and blue spectral range. Pulse energies are stable on a 1.1% level, averaged over 100 000 shots. By the use of conventional optics only, we compress a 0.2 µJ pulse at a center wavelength of 475 nm to a pulse duration of 12 fs, as measured by X-FROG. We test the capabilities of the approach by employing the ASHG-pulses as a pump in a transient absorption experiment on ß-carotene in solution.
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BACKGROUND: The AOK-proReha treatment approach determines the therapeutic measures during rehabilitation that should be taken in patients who have undergone total hip and total knee replacement surgery. The aim of our study is to scientifically assess the success of AOK-proReha in the routine care. METHODOLOGY: N=619 patients were surveyed about their health status at 3 time points. A historical control group was formed by referring to data of the QS-Reha(®) program and applying the propensity score matching method. RESULTS: We observed clear effects immediately after rehabilitation that revealed their maximum strength rising in most dimensions at the follow-up time point. Comparison with the control group demonstrated similar results in many dimensions. Some results were significantly better while none were significantly worse. CONCLUSION: The results of the AOK proReha approach are promising. Under appropriate conditions, the propensity score matching method is a sensible alternative to conducting a randomized controlled trial.
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Artroplastia de Quadril/reabilitação , Artroplastia do Joelho/reabilitação , Interpretação Estatística de Dados , Avaliação de Resultados em Cuidados de Saúde/métodos , Pontuação de Propensão , Idoso , Feminino , Alemanha , Humanos , Masculino , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The correction of genetic mutations by homologous recombination is an attractive approach to gene therapy. We used the DNA double-strand breaks introduced by the site-specific endonuclease I-Sce1 as a means of increasing homologous recombination of an exogenous DNA template in murine hematopoietic stem cells (mHSCs). To develop this approach, we chose an Artemis knockout (Art(-/-)) mouse in which exon 12 of the Artemis gene had been replaced by an I-Sce1 recognition site. The I-Sce1 enzyme and the Artemis correction template were each delivered by a self-inactivating (SIN)-integrase-defective lentiviral vector (SIN-IDLV-CMV-ISce1 and SIN-IDLV-Art, respectively). Transduction of Art(-/-) mHSCs with the two vectors successfully reverted the Art(-/-) phenotype in 2 of our 10 experiments. Even though the potential for genotoxicity has yet to be evaluated, this new approach to gene editing appears to be promising. Improving the efficacy of this approach will require further technical work.
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Endonucleases/genética , Terapia Genética/métodos , Células-Tronco Hematopoéticas/citologia , Recombinação Homóloga/genética , Proteínas Nucleares/genética , Animais , Quebras de DNA de Cadeia Dupla , Reparo do DNA , Desoxirribonucleases de Sítio Específico do Tipo II , Endonucleases/deficiência , Vetores Genéticos , Lentivirus/genética , Camundongos , Camundongos Knockout , Mutação , Proteínas Nucleares/deficiência , Proteínas de Saccharomyces cerevisiae , Transdução GenéticaRESUMO
We investigate the nature of the S* excited state in carotenoids by performing a series of pump-probe experiments with sub-20 fs time resolution on spirilloxanthin in a polymethyl-methacrylate matrix varying the sample temperature. Following photoexcitation, we observe sub-200 fs internal conversion of the bright S2 state into the lower-lying S1 and S* states, which in turn relax to the ground state on a picosecond time scale. Upon cooling down the sample to 77 K, we observe a systematic decrease of the S*/S1 ratio. This result can be explained by assuming two thermally populated ground state isomers. The higher lying one generates the S* state, which can then be effectively frozen out by cooling. These findings are supported by quantum chemical modeling and provide strong evidence for the existence and importance of ground state isomers in the photophysics of carotenoids.
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Modelos Teóricos , Temperatura , Isomerismo , Luz , Nitrogênio/química , Polimetil Metacrilato/química , Teoria Quântica , Análise Espectral , Xantofilas/químicaRESUMO
AIMS: The demands being made on quality assurance and quality management in ambulatory care nursing and inpatient nursing facilities continue to grow. As opposed to health-care facilities such as hospitals and rehabilitation centres, we know of no other empirical studies addressing the current state of affairs in quality management in nursing institutions. The aim of this investigation was, by means of a questionnaire, to analyse the current (as of spring 2011) dissemination of quality management and certification in nursing facilities using a random sample as representative as possible of in- and outpatient institutions. METHOD: To obtain our sample we compiled 800 inpatient and 800 outpatient facilities as a stratified random sample. Federal state, holder and, for inpatient facilities, the number of beds were used as stratification variables. 24% of the questionnaires were returned, giving us information on 188 outpatient and 220 inpatient institutions. While the distribution in the sample of outpatient institutions is equivalent to the population distribution, we observed discrepancies in the inpatient facilities sample. As they do not seem to be related to any demonstrable bias, we assume that our data are sufficiently representative. RESULTS: 4 of 5 of the responding facilities claim to employ their own quality management system, however the degree to which the quality management mechanisms are actually in use is an estimated 75%. Almost 90% of all the facilities have a quality management representative who often possesses specific additional qualifications. Many relevant quality management instruments (i. e., nursing standards of care, questionnaires, quality circles) are used in 75% of the responding institutions. Various factors in our data give the impression that quality management and certification efforts have made more progress in the inpatient facilities. Although 80% of the outpatient institutions claim to have a quality management system, only 32.1% of them admit to having already been (or be in current preparation to be) certified, a figure that was 41.5% among the inpatient facilities. These percentages are smaller when one relies on information provided by the certifying institutions themselves rather on the nursing facilities. Most frequent is the certification according to the DIN EN ISO 9001 standard, since the care-specific certification procedures most widespread on the market enable facilities to combine a care-specific certificate with one according to DIN norms. CONCLUSION: Quality management has become very widespread in nursing facilities: every third institution claims to have been certified, and the trend to become certified has clearly intensified over the last few years. We observe overall very great acceptance of both internal quality management and external quality assurance. We suspect that the current use of quality management instruments in many nursing facilities will not fall behind such efforts in hospitals and rehabilitation centres.
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Assistência Ambulatorial/estatística & dados numéricos , Serviço Hospitalar de Enfermagem/estatística & dados numéricos , Enfermagem Ambulatorial/estatística & dados numéricos , Garantia da Qualidade dos Cuidados de Saúde/organização & administração , Gestão da Qualidade Total/organização & administração , Alemanha , Garantia da Qualidade dos Cuidados de Saúde/métodos , Gestão da Qualidade Total/métodosRESUMO
The kinetics of electrogenic events associated with the different steps of the light-induced proton pump of bacteriorhodopsin is well studied in a wide range of time scales by direct electric methods. However, the investigation of the fundamental primary charge translocation phenomena taking place in the functional energy conversion process of this protein, and in other biomolecular assemblies using light energy, has remained experimentally unfeasible because of the lack of proper detection technique operating in the 0.1- to 20-THz region. Here, we show that extending the concept of the familiar Hertzian dipole emission into the extreme spatial and temporal range of intramolecular polarization processes provides an alternative way to study ultrafast electrogenic events on naturally ordered biological systems. Applying a relatively simple experimental arrangement based on this idea, we were able to observe light-induced coherent terahertz radiation from bacteriorhodopsin with femtosecond time resolution. The detected terahertz signal was analyzed by numerical simulation in the framework of different models for the elementary polarization processes. It was found that the principal component of the terahertz emission can be well described by excited-state intramolecular electron transfer within the retinal chromophore. An additional slower process is attributed to the earliest phase of the proton pump, probably occurring by the redistribution of a H bond near the retinal. The correlated electron and proton translocation supports the concept, assigning a functional role to the light-induced sudden polarization in retinal proteins.
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Bacteriorodopsinas/química , Elétrons , Halobacterium salinarum/química , Prótons , Radiação , Simulação por Computador , Transporte de Elétrons , Cinética , Óptica e Fotônica , Bombas de Próton , Fatores de TempoRESUMO
Exciton-exciton annihilation (EEA) is a ubiquitous phenomenon, which may limit the efficiency of photovoltaic devices. Conventional methods of determining EEA time scales rely on measuring the intensity dependence of third-order signals. In this work, we directly extract the annihilation rate of molecular excitons in a covalently joined molecular trimer without the need to perform and analyze intensity dependent data by employing fifth-order coherent optical spectroscopy signals emitted into ±2kâ1 ∓ 2kâ2 + kâ3 phase matching directions. Measured two-dimensional line shapes and their time traces are analyzed in the framework of the many-body version of the Frenkel exciton model, extended to incorporate annihilation dynamics. Combining double-sided Feynman diagrams with explicit simulations of the fifth-order response, we identify a single peak as a direct reporter of EEA. We retrieve an annihilation time of 30 fs for the investigated squaraine trimer.
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Time-resolved photoemission with ultrafast pump and probe pulses is an emerging technique with wide application potential. Real-time recording of nonequilibrium electronic processes, transient states in chemical reactions, or the interplay of electronic and structural dynamics offers fascinating opportunities for future research. Combining valence-band and core-level spectroscopy with photoelectron diffraction for electronic, chemical, and structural analyses requires few 10 fs soft X-ray pulses with some 10 meV spectral resolution, which are currently available at high repetition rate free-electron lasers. We have constructed and optimized a versatile setup commissioned at FLASH/PG2 that combines free-electron laser capabilities together with a multidimensional recording scheme for photoemission studies. We use a full-field imaging momentum microscope with time-of-flight energy recording as the detector for mapping of 3D band structures in (kx, ky, E) parameter space with unprecedented efficiency. Our instrument can image full surface Brillouin zones with up to 7 Å-1 diameter in a binding-energy range of several eV, resolving about 2.5 × 105 data voxels simultaneously. Using the ultrafast excited state dynamics in the van der Waals semiconductor WSe2 measured at photon energies of 36.5 eV and 109.5 eV, we demonstrate an experimental energy resolution of 130 meV, a momentum resolution of 0.06 Å-1, and a system response function of 150 fs.
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The Djungarian hamsters of our breeding colony show unstable daily activity patterns when kept under standard laboratory conditions. Moreover, part of them develops a delayed activity onset (DAO) or an arrhythmic phenotype. In former studies, we have shown that the system of photic entrainment works at its limits. If the period length (tau) increases, which is the case in DAO hamsters, the light-induced phase advances are too small to compensate the daily delay of the activity rhythm caused by tau being longer than 24 h. Accordingly, under natural conditions, there must be further (environmental) factors to enable a stable entrainment. One of these may be the higher level of motor activity. Animals must cover long distances to search for food, sexual partners and others. In the laboratory, hamsters are kept singly in small cages. This does restrict animals' options for motor activity. Also, there is less need for moving around as the hamsters are fed ad libitum. In the present study, a series of experiments was performed to investigate the putative effect of the activity level. To begin with, wild type (WT) and DAO animals were given access to running wheels. 50% of DAO hamsters developed a WT activity pattern. As the main reason for the DAO phenomenon is their long tau together with a too weak photic phase response, the effect of wheel running on these parameters was investigated in further experiments. With higher activity level, tau decreased in WT hamsters but increased in DAO animals even though the increase for the activity onset was only close to significance. Moreover, the photic phase responses were weaker though significant only for the activity offset of DAO hamsters. Based on the assumptions that running wheel activity will affect the phase response and/or the free running period, the results of the present paper do not provide an explanation for why part of DAO hamsters developed a WT phenotype when they had access to running wheels. Obviously, mechanisms downstream from the suprachiasmatic nuclei must be taken into account when investigating the stabilizing, improving circadian entrainment effect of motor activity.
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Comportamento Animal , Ritmo Circadiano , Cricetinae/fisiologia , Ciência dos Animais de Laboratório , Fotoperíodo , Condicionamento Físico Animal , Animais , LuzRESUMO
OBJECTIVE: To determine whether changes in hand skin blood flow in diabetic men could be demonstrated with liquid crystal contact thermography and to assess the relative effects of autonomic neuropathy and microangiopathy on these changes. RESEARCH DESIGN AND METHODS: Thirty-four diabetic and 12 age-matched nondiabetic men comprised the study. The diabetic men were categorized according to standard cardiovascular autonomic function tests and the presence or absence of background or proliferative retinopathy and/or proteinuria. Bilateral hand thermograms were measured at rest and after immersion of the right hand in ice-cold water. RESULTS: Diabetic men with definite or severe autonomic neuropathy (n = 13) had a high frequency of anisothermal baseline thermograms (77 vs. 25% in nondiabetic subjects, P less than 0.05). After ice-cold water immersion, right-hand recovery was abnormally slow (514 +/- 157 arbitrary U, area under the curve) compared with nondiabetic men (685 +/- 135 arbitrary U, P less than 0.01). Diabetic men with proliferative retinopathy (n = 8) all had definite or severe autonomic neuropathy and showed the same abnormalities. Diabetic men with nor or early autonomic changes showed normal thermographic patterns. CONCLUSIONS: These results are consistent with increased palmar arteriovenous shunt blood flow or capillary closure in the hands of diabetic patients with definite or severe autonomic neuropathy. They indicate that thermoregulatory reflex changes in hand skin blood flow are controlled by the autonomic nervous system. It is possible, however, that diabetic microangiopathy, associated with the presence of proliferative retinopathy, also independently affects hand skin blood flow.
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Angiopatias Diabéticas/fisiopatologia , Neuropatias Diabéticas/fisiopatologia , Pele/irrigação sanguínea , Adulto , Retinopatia Diabética/fisiopatologia , Mãos , Humanos , Masculino , Pessoa de Meia-Idade , Fluxo Sanguíneo Regional , TermografiaRESUMO
The genus Phodopus consists of three species--P. campbelli (Pc), P. sungorus (Ps), and P. roborovskii (Pr). They inhabit steppes, semi-deserts, and deserts in continental Asia with a climate changing from a moderate to a hard Continental one with extreme daily and seasonal variations. These different environmental challenges are likely to have consequences for hamsters' morphology, physiology, and behavior. Hamsters of all three species were investigated during the course of the year in the laboratory though using natural lighting and temperature conditions. Motor activity and body temperature were measured continuously, and body mass, testes size, and fur coloration every 1-2 weeks. With regard to the pattern of activity, nearly twice as many Pc as Ps hamsters (25 vs. 14%) failed to respond to changes of photoperiod, whereas all Pr hamsters did. Body mass and testes size were high in summer and low in winter, with the biggest relative change in Ps and the lowest in Pr hamsters. Changes of fur coloration were found in Ps hamsters only. All responding animals (that is excluding Pr), exhibited regular torpor bouts during the short winter days. In autumn, seasonal changes started considerably earlier in Ps hamsters. To investigate the putative causes of these different time courses, a further experiment was performed, to identify the critical photoperiod. Hamsters were kept for 10 weeks under different photoperiods, changing from 16 to 8 h light per day. Motor activity was recorded continuously, to identify responding and non-responding animals. Body mass was measured at the beginning and the end of the experiment, testes mass only at the end. The critical photoperiod was found to be similar in all three species. Though in a further experiment, Pc and Pr hamsters showed a delayed response, whereas the changes in Ps hamsters started immediately following transfer to short-day conditions. The results show that interspecific differences in seasonal adaptation exist, even between the closely related Ps and Pc hamsters, possibly due to different conditions in their natural habitat. Also, the impact of environmental factors like climatic conditions and food resources may differ between species.
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Ecossistema , Phodopus/fisiologia , Estações do Ano , Ciclos de Atividade , Adaptação Fisiológica , Animais , Regulação da Temperatura Corporal , Peso Corporal , Cricetinae , Cabelo , Cor de Cabelo , Masculino , Atividade Motora , Tamanho do Órgão , Phodopus/crescimento & desenvolvimento , Fotoperíodo , Especificidade da Espécie , Temperatura , Testículo/crescimento & desenvolvimento , Fatores de TempoRESUMO
The heat stable inhibitor of cAMP-dependent protein kinase (PKIalpha) contains both a nuclear export signal (NES) and a high affinity inhibitory region that is essential for inhibition of the catalytic subunit of the kinase. These functions are sequentially independent. Two-dimensional NMR spectroscopy was performed on uniformly [15N]-labeled PKIalpha to examine its structure free in solution. Seventy out of 75 residues were identified, and examination of the CaH chemical shifts revealed two regions of upfield chemical shifts characteristic of alpha-helices. When PKIalpha was fragmented into two functionally distinct peptides for study at higher concentrations, no significant alterations in chemical shifts or secondary structure were observed. The first ordered region, identified in PKIalpha (1-25), contains an alpha-helix from residues 1-13. This helix extends by one turn the helix observed in the crystal structure of a PKIalpha (5-24) peptide bound to the catalytic subunit. The second region of well-defined secondary structure, residues 35-47, overlaps with the nuclear export signal in the PKIalpha (26-75) fragment. This secondary structure consists of a helix with a hydrophobic face comprised of Leu37, Leu41, and Leu44, followed by a flexible turn containing Ile46. These four residues are critical for nuclear export function. The remainder of the protein in solution appears relatively unstructured, and this lack of structure surrounding a few essential and well-defined signaling elements may be characteristic of a growing family of small regulatory proteins that interact with protein kinases.
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Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Núcleo Celular/metabolismo , Sequência de Aminoácidos , Transporte Biológico , Domínio Catalítico , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Dados de Sequência Molecular , Fragmentos de Peptídeos/metabolismo , Estrutura Secundária de Proteína , Relação Estrutura-AtividadeRESUMO
Specific cytotoxicity of human CD56+NK and LAK cells was quantitatively inhibited by acetylated mannose, galactose and glucose (Scand. J. Immunol., in press). The respective NK cell receptors were isolated from human leukocyte lysates by affinity chromatography based on 60% deacetylated penta-acetates of mannose, galactose and glucose. All three affinity isolates contained a main component with +/- 87 kDa molecular mass exhibiting about the same patterns of isoforms at pI 4.90, 4.75, 4.60 and 4.50 in isoelectric focusing. Moreover, preincubation of tumor target cells with the three 87-kDa receptors revealed very similar inhibitory potentials for human NK and LAK cytotoxicity showing dose-dependent inhibition between 20 (no inhibition) and 700 pmol/ml (100% inhibition) receptor concentration. The data support the assumption that the three affinity isolates contain the same type of receptor directed against a unique epitope common to acetylated mannose, galactose and glucose.
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Antígeno CD56/imunologia , Hexoses/metabolismo , Células Matadoras Ativadas por Linfocina/imunologia , Células Matadoras Naturais/imunologia , Receptores de Superfície Celular/metabolismo , Ligação Competitiva , Cromatografia de Afinidade , Citotoxicidade Imunológica , Humanos , Leucócitos/química , Ligação Proteica/imunologiaRESUMO
Target cell recognition and cytotoxicity of human CD56+ NK and LAK cells is readily inhibited by acetylated mannose. Two respective NK cell receptor candidates were isolated from human leukocyte lysates by mannose acetate affinity chromatography. The 87-kDa receptor showed sequence homologies with lactoferrin and the 59-kDa receptor represented a complex of two Ca-binding proteins MRP-8 and MRP-14 reportedly expressed only by cells of myeloid origin. The 87-kDa receptor exhibited heterogeneity in isoelectric focusing and behaved entirely differently from lactoferrin. Preincubation of tumor target cells with the 87-kDa receptor inhibited competitively target cell recognition and cytotoxicity of human CD56+ NK and LAK cells.
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Células Matadoras Ativadas por Linfocina/imunologia , Células Matadoras Naturais/imunologia , Lectinas Tipo C , Lectinas de Ligação a Manose , Receptores de Superfície Celular/imunologia , Receptores de Superfície Celular/isolamento & purificação , Acetilação , Sequência de Aminoácidos , Antígenos de Diferenciação/química , Proteínas de Ligação ao Cálcio/química , Calgranulina A , Calgranulina B , Citotoxicidade Imunológica , Eletroforese em Gel de Poliacrilamida , Humanos , Lactoferrina/química , Receptor de Manose , Dados de Sequência Molecular , Peso Molecular , Receptores de Superfície Celular/química , Homologia de Sequência de Aminoácidos , Células Tumorais CultivadasRESUMO
In a randomized prospective study of patients having cardiac surgery, autologous blood collected from mediastinal tubes was autotransfused preferentially in 63 patients (ATS), whereas 51 patients received bank blood for transfusion (control). Comparison of the two groups showed no significant difference in regard to age, sex, operations performed, or total postoperative bleeding (ATS 813 +/- 121 ml. per square meter versus control 711 +/- 93 ml. per square meter; N.S.) Although mean postoperative blood replacement was similar in the two groups (ATS 4.3 +/- 0.6 units per patient versus control 4.8 +/- 0.6 units per patient), requirements for transfusion of stored bank blood were reduced by 50 percent in the ATS group (ATS 2.4 +/- 0.3 units per patient versus control 4.8 +/- 0.6 units per patient; p less than 0.005). Coagulation studies demonstrated that this blood was defibrinogenated; yet it contains significantly more platelets and clotting factors than does bank blood. In this study, autotransfusion of shed mediastinal blood was safe and simple. It significantly reduced bank blood requirements and resulted in substantial financial savings for the patients and the hospital.
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Transfusão de Sangue Autóloga , Procedimentos Cirúrgicos Cardíacos , Contagem de Células Sanguíneas , Plaquetas , Transfusão de Sangue , Procedimentos Cirúrgicos Cardíacos/mortalidade , Coagulação Intravascular Disseminada/etiologia , Feminino , Hemorragia/etiologia , Humanos , Masculino , Maryland , Mediastino , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Estudos ProspectivosRESUMO
A case report of a patient with Marfan's syndrome who developed acute supravalvular aortic stenosis following aortic valve replacement and reconstruction of the ascending aorta is presented. Early recognition of this complication permitted reoperation and successful repair. One similar case has been reported previously.
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Aorta/cirurgia , Estenose da Valva Aórtica/etiologia , Valva Aórtica/cirurgia , Próteses Valvulares Cardíacas , Síndrome de Marfan/cirurgia , Complicações Pós-Operatórias , Doença Aguda , Adulto , Humanos , MasculinoRESUMO
An autotransfusion technique has been developed for collection and reinfusion of shed mediastinal blood. This system has been routinely applied in the postoperative management of 592 consecutive adult and 108 pediatric cardiac surgical patients. Two hundred seventy-one adult patients (46%) and thirty-six pediatric patients (33%) actually received autologous blood. Autotransfusion volume ranged from 50 to 21,350 ml per patient. In 1976 at our institution, homologous transfusion requirements averaged 8.4 +/- 0.7 units per adult patient. During 1978, with the routine use of postoperative autotransfusion, bank blood transfusions were lowered to 4.2 +/- 0.3 units per patient (p less than 0.001). In contrast to perioperative autotransfusion techniques, collection and reinfusion of shed mediastinal blood is particularly useful for intravascular volume replacement in patients with serious postoperative bleeding.
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Transfusão de Sangue Autóloga/métodos , Procedimentos Cirúrgicos Cardíacos , Adulto , Coleta de Amostras Sanguíneas/instrumentação , Transfusão de Sangue Autóloga/economia , Transfusão de Sangue Autóloga/instrumentação , Procedimentos Cirúrgicos Cardíacos/economia , Criança , Ponte de Artéria Coronária , Custos e Análise de Custo , Equipamentos Descartáveis , Aneurisma Cardíaco/cirurgia , Doenças das Valvas Cardíacas/cirurgia , Próteses Valvulares Cardíacas , Humanos , Mediastino , Métodos , Cuidados Pós-OperatóriosRESUMO
In this work we analyze how nuclear coherences modulate diagonal and off-diagonal peaks in two-dimensional electronic spectroscopy. 2D electronic spectra of pinacyanol chloride are measured with 8 fs pulses, which allows coherent excitation of the 1300 cm(-1) vibrational mode. The 2D spectrum reveals both diagonal and off-diagonal peaks related to the vibrational mode. On early time scales, up to 30 fs, coherent dynamics give rise to oscillations in the amplitudes, positions, and shapes of the peaks in the 2D spectrum. We find an anticorrelation between the amplitude and the diagonal width of the two diagonal peaks. The measured data are reproduced with a model incorporating a high frequency mode coupled to an electronic two-level-system. Our results show that these anticorrelated oscillations occur for vibrational wavepackets and not exclusively for electronic coherences as has been assumed previously.