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Biomedical subjects

M Neu

Publications and source records attributed to M Neu.

At least 19 recordsLinked to original sources
PubMed2005

Nano-carriers for DNA delivery to the lung based upon a TAT-derived peptide covalently coupled to PEG-PEI.

Gene therapy aimed at the respiratory epithelium holds therapeutic potential for diseases such as cystic fibrosis and lung cancer. Polyethylenimine (PEI) has been utilized for gene delivery to the airways. In this study, we describe a new modification of PEI, in which an oligopeptide related to the protein transduction domain of HIV-1 TAT was covalently coupled to 25 kDa PEI (PEI) through a heterobifunctional polyethylenglycol (PEG) spacer resulting in a TAT-PEG-PEI conjugate. Improved DNA reporter gene complexation and protection was observed for small (approximately 90 nm) polyplexes as well as significantly improved stability against polyanions, Alveofact, bronchial alveolar lining fluid and DNase. To determine polyplex toxicity in vitro, MTT assays were performed and, for in vivo testing, the mice bronchial alveolar lavage was investigated for total cell counts, quantity of neutrophils, total protein and TNF-alpha concentration. All parameters suggest significantly lower toxicity for TAT-PEG-PEI. Transfection efficiencies of both PEI and TAT-PEG-PEI polyplexes with DNA were studied under in vitro conditions (A549) and in mice after intratracheal instillation. While luciferase expression in A549 cells was much lower for TAT-PEG-PEI (0.2 ng/mg protein) than for PEI (2 ng/mg), significantly higher transfection efficiencies for TAT-PEG-PEI were detected in mice. Reporter gene expression was distributed through bronchial and alveolar tissue. Thus, TAT-PEG-PEI represents a new approach to non-viral gene carriers for lung therapy, comprising protection for plasmid DNA, low toxicity and significantly enhanced transfection efficiency under in vivo conditions.

Animals↗
PubMed2002

Unusual craniocerebral injury caused by a pneumatic nail gun.

A man was found unconscious near a ladder in a house. After resuscitation he was brought to a hospital and X-rays of the skull showed that two 12-cm long nails had completely penetrated the cranial cavity. The nails were operatively removed and after treatment for 5 weeks, the patient was transferred to a rehabilitation centre with a decreasing hemiparesis on the left side and general deterioration and then, after an attempted suicide to a psychiatric hospital. The perforating cranio-cerebral injury from a pneumatic nail gun known to reach only low muzzle velocities is a very unusual finding.

Accidents, Occupational↗
PubMed2001

The crystal and molecular structures of diferric porcine and rabbit serum transferrins at resolutions of 2.15 and 2.60 A, respectively.

The serum transferrins are monomeric proteins with a molecular mass of around 80 kDa and are responsible for the transport of iron in vertebrates. The three-dimensional structures of diferric porcine and rabbit serum transferrin have been refined against X-ray diffraction data extending to 2.15 and 2.60 A, respectively. Data for both proteins were collected using synchrotron radiation at temperatures of 277 K. The porcine protein crystallizes in the space group C2, with unit-cell parameters a = 223.8, b = 44.9, c = 78.9 A, beta = 105.4 degrees with one molecule in the asymmetric unit. The structure was solved by molecular-replacement methods using rabbit serum transferrin as the search model. The structure was refined using REFMAC, with a final residual of 13.8% (R(free) = 18.2% for a 5% data sample) for all data to 2.15 A. The final model comprises 5254 protein atoms, two Fe(3+) cations and two CO(3)(2-) anions, one N-acetyl glucosamine moiety and 494 water molecules. The rabbit protein crystallizes in space group P4(3)2(1)2, with unit-cell parameters a = 127.2, c = 144.9 A and one molecule per asymmetric unit. The structure was solved using the method of multiple isomorphous replacement and refined using REFMAC to give a final residual of 18.6% (R(free) = 22.2% for a 5% data sample) for all data to 2.60 A. The final model comprises 5216 protein atoms, two Fe(3+) cations and two CO(3)(2-) anions, a Cl(-) anion and 206 solvent molecules; there is no clear indication of the carbohydrate moiety attached to Asn490 (rabbit serum numbering). Both molecules adopt a bilobal structure typical for members of the transferrin family. Each of the structurally homologous lobes contains two dissimilar domains with a single iron-binding site buried within the interdomain cleft. The porcine serum protein lacks an interdomain disulfide bridge close to the connecting peptide between the lobes, but this seems to have little effect on the overall orientation of the lobes. The N-lobes of both proteins possess lysine residues, one from each of the two domains, that lie in close proximity to one another to form the so-called dilysine trigger. The more acid-labile release of iron from serum transferrins than from lactoferrins is discussed.

Animals↗
PubMed2001

Clinical and neurophysiological observations after superior petrosal vein obstruction during surgery of the cerebellopontine angle: case report.

IMPORTANCE: Preservation of venous drainage during surgery of the cerebellopontine angle has received little attention. CLINICAL PRESENTATION: We describe changes in brainstem auditory evoked potentials after temporary obstruction of the superior petrosal vein during surgical resection of a small meningioma at the petrous apex via a standard suboccipital-lateral approach. Temporary clipping of the petrosal vein resulted in deterioration of the brainstem auditory evoked potentials. The tumor was removed with preservation of the superior petrosal vein. CONCLUSION: A transient postoperative cochlear nerve deficit emphasizes the importance of venous drainage and its preservation during surgery for small lesions of the cerebellopontine angle that do not distort normal anatomic structures.

Cerebellopontine Angle↗
PubMed2001

Generalizability and clinical utility of a practice-based infant pain assessment instrument.

The purpose of this study was to determine the clinical usefulness and generalizability of an infant pain assessment instrument. Earlier work showed that this instrument--an algorithm derived from a model of infant pain assessment-possessed excellent content validity, criterion-like validity, and 3-month stability (test-retest reliability). In this study, generalizability was determined by comparing the percentage agreement between inexperienced pediatric nurses and one author, both using the tool to assess pain of infants in various clinical settings, and by comparing the percentage agreement between one author who used the tool to assess pain and the infant's pediatric nurse caretaker who used his or her clinical expertise, not the tool, to assess pain across various clinical settings. The results show excellent generalizability.

Humans↗
PubMed2001

Vasoactive treatment for hearing preservation in acoustic neuroma surgery.

OBJECT: Delayed hearing loss following surgery for acoustic neuroma indicates anatomical and functional preservation of the cochlear nerve and implies that a pathophysiological mechanism is initiated during surgery and continues thereafter. Intraoperative brainstem auditory evoked potentials (BAEPs) typically demonstrate gradual reversible loss of components in these patients. METHODS: Based on this BAEP pattern, a consecutive series of 41 patients with unilateral acoustic neuromas was recruited into a prospective randomized study to investigate hearing outcomes following the natural postoperative course and recuperation after vasoactive medication. Both groups were comparable in patient age, tumor size, and preoperative hearing level. Twenty patients did not receive postoperative medical treatment. In 70% of these patients anacusis was documented and in 30% hearing was preserved. Twenty-one patients were treated with hydroxyethyl starch and nimodipine for an average of 9 days. In 66.6% of these patients hearing was preserved and in 33.3% anacusis occurred. CONCLUSIONS: These results are statistically significant (p < 0.05, chi2 = 5.51) and provide evidence that these surgically treated patients suffer from a disturbed microcirculation that causes delayed hearing loss following removal of acoustic neuromas.

Adult↗
PubMed2000

Validity and reliability of a practice-based infant pain assessment instrument.

The purpose of this study was to determine the validity and reliability of an infantpain assessment instrument. This instrument is an algorithm derived from a model of infant pain assessment that emerged from interviews with pediatric nurses and includes methods for evaluating clinical data that suggest the Iikelihood of pain. Unlike other instruments, this instrument is not restricted to use in clinical situations where the likelihood of pain is evident. This instrument determines the three levels of pain that guide subsequent nursing interventions: no pain, mild pain (discomfort), and severe pain. Content validity was determined by obtaining and incorporating feedback from a focus group of practicing pediatric nurses on various developmental stages of the instrument. Criterion-like validity was excellent: Mean Pearson correlations between the pain ratings of 18 videotaped infants by 24 nursing students using the instrument with ratings performed by a panel of 5 expert nurses not using any instrument was 0.96. Test-retest reliability, as measured by Pearson correlations between pain ratings 3 months apart, was also excellent (r = 0.91).

Humans↗
PubMed1999

X-ray crystal structure of human dopamine sulfotransferase, SULT1A3. Molecular modeling and quantitative structure-activity relationship analysis demonstrate a molecular basis for sulfotransferase substrate specificity.

Humans are one of the few species that produce large amounts of catecholamine sulfates, and they have evolved a specific sulfotransferase, SULT1A3 (M-PST), to catalyze the formation of these conjugates. An orthologous protein has yet to be found in other species. To further our understanding of the molecular basis for the unique substrate selectivity of this enzyme, we have solved the crystal structure of human SULT1A3, complexed with 3'-phosphoadenosine 5'-phosphate (PAP), at 2.5 A resolution and carried out quantitative structure-activity relationship (QSAR) analysis with a series of phenols and catechols. SULT1A3 adopts a similar fold to mouse estrogen sulfotransferase, with a central five-stranded beta-sheet surrounded by alpha-helices. SULT1A3 is a dimer in solution but crystallized with a monomer in the asymmetric unit of the cell, although dimer interfaces were formed by interaction across crystallographic 2-fold axes. QSAR analysis revealed that the enzyme is highly selective for catechols, and catecholamines in particular, and that hydrogen bonding groups and lipophilicity (cLogD) strongly influenced K(m). We also investigated further the role of Glu(146) in SULT1A3 using site-directed mutagenesis and showed that it plays a key role not only in defining selectivity for dopamine but also in preventing many phenolic xenobiotics from binding to the enzyme.

Alanine↗
PubMed1999

The prognostic value of intraoperative BAEP patterns in acoustic neurinoma surgery.

Based on a consecutive series of 70 hearing patients with unilateral acoustic neurinomas and intraoperative monitoring of brain-stem auditory evoked potentials (BAEP), 4 dynamic BAEP patterns could be characterized. These patterns correspond with early and late postoperative hearing outcome. All patients with stable wave V (pattern 1) showed definite hearing preservation, all patients with irreversible abrupt loss of BAEP (pattern 2) lost their hearing, despite early hearing preservation in two cases. All patients with irreversible progressive loss of either wave I or wave V (pattern 3) eventually suffered from definite postoperative hearing loss, despite early hearing preservation in two cases. Those cases with intraoperative reversible loss of BAEP (pattern 4) showed variable short and long term hearing outcome. In 34% hearing was preserved, 44% suffered from postoperative hearing loss, the remaining 22% showed postoperative hearing fluctuation, either as a delayed hearing loss or as reversible hearing loss. Postoperative hearing fluctuation indicates anatomical and functional preservation of the cochlear nerve during surgery and is suggestive of a pathophysiological mechanism initiated during the surgical procedure and continuing thereafter. Patients at risk for delayed hearing loss can be identified during surgery by a characteristic BAEP pattern and may benefit from vasoactive treatment.

Audiometry, Pure-Tone↗
PubMed1999

Testing a model of the nursing assessment of infant pain.

The purpose of this study was to test whether elements of an infant pain assessment model interacted as postulated by the model. The elements are the infant's response to comfort measures and the principle of consolability. Four different scenarios for each of 16 videotaped infants were prepared. Each scenario represented one of four different combinations of likelihood of pain and consolability and consisted of a videotape plus written clinical information. Forty-eight volunteer pediatric nurses assessed infant pain of 16 scenarios, each depicting one of the 16 infants. Mean level of assessed pain was highest for the "high likelihood of pain and difficult to console" group, second highest for the "high likelihood of pain and easily consoled" group, third highest for the "low likelihood of pain and difficult to console" group and least for the "low likelihood of pain and easily consoled" group. Findings supported the infant pain assessment model.

Female↗
PubMed1999

The influence of background clinical data on infant pain assessments.

The purpose of this study was to determine the importance of knowledge of clinical background data on nursing assessments of infant pain. In a quasi-experimental design, the infant pain assessments of two groups of pediatric nurses were compared. Both groups assessed the levels of pain of the same videotaped infants. One group also read clinical background data about each infant; the other group did not. The group who read clinical background data evidenced higher mean levels of assessed pain per videotaped infant than did the group who only viewed the videotapes. Findings underscore the importance of clinical data and clinical context in the process of assessing infant pain and the risk of underestimating pain when all factors are not considered.

Age Factors↗
PubMed1998

The nature of ligand-induced conformational change in transferrin in solution. An investigation using X-ray scattering, XAFS and site-directed mutants.

Ligand-induced conformational change in transferrins has been studied by site-directed mutagenesis of human serum half molecule (N-lobe), X-ray absorption fine structure (XAFS) spectroscopy and X-ray solution scattering. Use of recent advances in data analysis has been made for extracting model-independent molecular shapes from X-ray solution scattering data for the intact, the half molecule and its mutants. Clear evidence is provided that the transferrin molecule (intact as well as N-lobe), in its apo and holo forms, exists for the majority of the time in well-defined specific conformations representing the "fully opened" and "closed" states of the molecule, respectively. Evidence is also provided for the existence of an additional conformation, referred to here as the "intermediate" conformation for simplicity, which is trapped in the case of some of the mutants in the iron-bound form. We suggest that domain closure in the transferrin molecule is a two-step process, with the intermediate conformation representing the first stage of domain closure (approximately 20 degrees hinge-twist of domain II). Our data are not inconsistent with the ligand-free molecule sampling the closed states occasionally (< or = 10%) but are not in support of a continuous conformational search between the fully opened and closed states in the absence of iron.

Animals↗
PubMed1998

Preservation of hearing in large acoustic neurinomas following removal via suboccipito-lateral approach.

A consecutive series of 61 patients with pre-operative hearing underwent surgical removal of a large acoustic neurinoma via the lateral suboccipital approach between 1984 and 1996. Brain-stem auditory evoked potentials (BAEP) were present in all cases before surgery and all patients underwent intra-operative monitoring of BAEP. The average tumour size including the portion within the meatus acusticus internus was 30, 5 mm (range 20 mm-49 mm). Complete tumour removal was achieved in all but three cases. In 43.1% of patients with complete tumour removal hearing was preserved initially after surgery. Delayed postoperative hearing loss was observed in 11 patients and hearing recovery in 2 patients leading to 27.5% definite hearing preservation. Hearing preservation was achieved in 37% of cases with tumour size between 20 mm-29 mm and in 23.5% of case with tumour size larger than 30 mm. These results indicate that even in large neurinoms hearing preservation should be attempted in all patients with documented pre-operative hearing and BAEP.

Adult↗
PubMed1997

Rab7: NMR and kinetics analysis of intact and C-terminal truncated constructs.

Rab proteins are a family of approximately 25kD ras-related GTPases which are associated with distinct intracellular membranes where they control vesicle traffic between intracellular compartments. The late-endosomal rab protein rab7(1-207), (lacking only the C-terminal lipids of the native molecule) and three C-terminal truncated constructs rab7(1-202), rab7(1-197) and rab7(1-182) were purified using an E. coli expression system. The C-terminal tail region of rab proteins is of special interest because it is thought to target rab proteins to particular intracellular membranes. A comparison of TOCSY-NMR spectra from intact rab7(1-207) and the tail-less construct rab7(1-182) suggested that much of the C-terminal tail is flexible in solution. The GTP hydrolysis, and GDP association and dissociation rates for all the truncated and intact constructs were similar, showing that the tail region of rab7(1-207) has little influence on the hydrolysis and exchange rates of the nucleotide.

GTP-Binding Proteins↗
PubMed1997

Rab7: crystallization of intact and C-terminal truncated constructs complexed with GDP and GppNHp.

The GTP/GDP conformational switch of members of the rab family of ras-related GTP-ases control specific intracellular vesicle transport pathways. We report the crystallization of the late-endosomal rab protein rab7, in both GTP and GDP conformations. X-ray data from crystals of rab7(1-207) GppNHp (i.e., intact rab7, without C-terminal bound lipid, complexed with a non-hydrolysable GTP analog), rab7(1-197)GppNHp and rab7(1-197)GDP were collected to 1.9A (0 degree C), 1.76A (100 degrees K) and 1.75A (100 degrees K) respectively. Rab7-GDP crystals diffract to at least 1.35A.

Crystallography, X-Ray↗
PubMed1995

Quick Fluorescence-EXAFS: an Improved Method for Collection of Conventional XAFS Data, an Improved Method for Collection of Conventional XAFS Data and for Studying Reaction Intermediates in Dilute Systems.

The quick EXAFS (QuEXAFS) technique provides an alternative way of recording X-ray absorption fine-structure (XAFS) data where the scan time is reduced by moving the monochromator at a constant angular speed and recording the data ;on the fly'. Results are presented to show that the use of fluorescence detection with QuEXAFS is eminently suitable for studying reactions in dilute systems such as metalloproteins at a sub-minute time scale. In addition, we show that the fluorescence-QuEXAFS technique can reduce the overall time for normal data collection by some 50% compared with conventional step-by-step scanning EXAFS using the same optical system, thus reducing the total X-ray exposures of the samples. The use of X-rays for studying in situ redox reactions is demonstrated.

Journal Article↗