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Biomedical subjects

S Brandt

Publications and source records attributed to S Brandt.

At least 37 records · Page 2Linked to original sources

Technical Advance: A rapid method for detection of plant gene transcripts from single epidermal, mesophyll and companion cells of intact leaves.

A protocol for the detection of gene transcripts from single plant cells in living, undamaged plant tissue is described. Samples of leaf epidermal, mesophyll and companion cells were extracted by using glass microcapillaries and directly subjected to RT-PCR without any purification steps nor time consuming construction of cDNA libraries. The procedure is not restricted to surface cells or outer cell layers. Even cells from the central region of leaves could be harvested. For identification, companion cells were labelled by expression of the green fluorescent protein under control of a companion cell specific promoter. The described method is applicable to a wide range of plants and genes with different expression levels.

Journal Article↗

Influenza A virus lacking the NS1 gene replicates in interferon-deficient systems.

The NS1 protein is the only nonstructural protein encoded by influenza A virus. It has been proposed that the NS1 performs several regulatory functions during the viral replication cycle, including the regulation of synthesis, transport, splicing, and translation of mRNAs. Through the use of reverse genetics, a viable transfectant influenza A virus (delNS1) which lacks the NS1 gene has been generated. Our results indicate that the NS1 of influenza A virus is an auxiliary (virulence) factor which plays a crucial role in inhibiting interferon-mediated antiviral responses of the host.

Animals↗

Photometry and spectroscopy of the GRB 970508 optical counterpart

An optical transient within the error box of the gamma ray burst GRB 970508 was imaged 4 hours after the event. It displayed a strong ultraviolet excess, and reached maximum brightness 2 days later. The optical spectra did not show any emission lines, and no variations on time scales of minutes were observed for 1 hour during the decline phase. According to the fireball and afterglow models, the intensity should rise monotonically before the observed optical maximum, but the data indicate that another physical mechanism may be responsible for the constant phase seen during the first hours after the burst.

Journal Article↗

A history of stress alters drought calcium signalling pathways in Arabidopsis.

Environmental stresses commonly encountered by plants lead to rapid transient elevations in cytosolic free calcium concentration ([Ca2+]cyt) (Bush, 1995; Knight et al., 1991). These cellular calcium (Ca2+) signals lead ultimately to the increased expression of stress-responsive genes, including those encoding proteins of protective function (Knight et al., 1996; Knight et al., 1997). The kinetics and magnitude of the Ca2+ signal, or 'calcium signature', differ between different stimuli and are thought to contribute to the specificity of the end response (Dolmetsch et al., 1997; McAinsh and Hetherington, 1998). We measured [Ca2+]cyt changes during treatment with mannitol (to mimic drought stress) in whole intact seedlings of Arabidopsis thaliana. The responses of plants which were previously exposed to osmotic and oxidative stresses were compared to those of control plants. We show here that osmotic stress-induced Ca2+ responses can be markedly altered by previous encounters with either osmotic or oxidative stress. The nature of the alterations in Ca2+ response depends on the identity and severity of the previous stress: oxidative stress pre-treatment reduced the mannitol-induced [Ca2+]cyt response whereas osmotic stress pretreatment increased the [Ca2+]cyt response. Therefore, our data show that different combinations of environmental stress can produce novel Ca2+ signal outputs. These alterations are accompanied by corresponding changes in the patterns of osmotic stress-induced gene expression and, in the case of osmotic stress pre-treatment, the acquisition of stress-tolerance. This suggests that altered Ca2+ responses encode a 'memory' of previous stress encounters and thus may perhaps be involved in acclimation to environmental stresses.

Aequorin↗

Identification of a K+ channel from potato leaves by functional expression in Xenopus oocytes.

Poly(A)+ mRNA was isolated from leaves of potato plants (Solanum tuberosum L. cv. Desiree) according to standard protocols. This poly(A)+ mRNA was injected via glass microcapillaries into oocytes that were surgically removed from the African clawed toad Xenopus laevis. As a control, oocytes were either injected with H2O or remained untreated. Three days after injection the oocytes were analyzed by two electrode voltage clamping. Current voltage analysis revealed that a K+ channel from potato was functionally expressed in injected oocytes. The identity of this K+ channel was confirmed by its substrate specificity and a shift in the reversal potential. In particular, when the outside K+ concentration was increased the reversal potential of poly(A)+ injected oocytes shifted to more positive values. Furthermore, K+ outward currents declined when the outside K+ concentration was raised from 0.1 to 100 mM. Inward currents increased with an elevation of the K+ concentration. Several pharmaceuticals were tested for their potential to block this K+ channel. As a result, the channel was completely blocked by BaCl2. A three state reaction kinetic model was used to simulate the currents through the K+ transport protein as function of the extracellular K+ concentration. In particular, the simulation revealed current voltage relations that exactly matched the measured ones. Saturation of current voltage curves emerged from the simulation as a consequence of high extracellular potassium concentration.

Animals↗

Transfectant influenza A viruses with long deletions in the NS1 protein grow efficiently in Vero cells.

We established a reverse genetics system for the nonstructural (NS) gene segment of influenza A virus. This system is based on the use of the temperature-sensitive (ts) reassortant virus 25A-1. The 25A-1 virus contains the NS gene from influenza A/Leningrad/134/57 virus and the remaining gene segments from A/Puerto Rico (PR)/8/34 virus. This particular gene constellation was found to be responsible for the ts phenotype. For reverse genetics of the NS gene, a plasmid-derived NS gene from influenza A/PR/8/34 virus was ribonucleoprotein transfected into cells that were previously infected with the 25A-1 virus. Two subsequent passages of the transfection supernatant at 40 degreesC selected viruses containing the transfected NS gene derived from A/PR/8/34 virus. The high efficiency of the selection process permitted the rescue of transfectant viruses with large deletions of the C-terminal part of the NS1 protein. Viable transfectant viruses containing the N-terminal 124, 80, or 38 amino acids of the NS1 protein were obtained. Whereas all deletion mutants grew to high titers in Vero cells, growth on Madin-Darby canine kidney (MDCK) cells and replication in mice decreased with increasing length of the deletions. In Vero cells expression levels of viral proteins of the deletion mutants were similar to those of the wild type. In contrast, in MDCK cells the level of the M1 protein was significantly reduced for the deletion mutants.

Animals↗

Nocturnal polyuria and antidiuretic hormone levels in spinal cord injury.

OBJECTIVE: To establish baseline ADH levels in spinal cord injury patients and to evaluate whether spinal cord patients have attenuation of diurnal variation of ADH similar to children with enuresis and elderly with nocturnal polyuria. DESIGN: Twenty-seven healthy quadriplegic patients, ASIA impairment scale A, were evaluated for serum ADH levels at night and during the day. MAIN OUTCOME MEASURES: Evaluation of whether bladder overdistention caused by attenuation of diurnal variation of ADH is responsible for the episodes of autonomic dysreflexia and recurrent urinary tract infections in spinal cord injury patients who are on intermittent catheterization for bladder management. RESULTS: A lack of diurnal variation of ADH in the subject population. CONCLUSIONS: A trial of desaminocystein-D-arginine vasopressin (DDAVP) should be considered for patients with established attenuation of the diurnal variation ADH.

Adult↗

ClC-6 and ClC-7 are two novel broadly expressed members of the CLC chloride channel family.

We cloned two novel members of the CLC chloride channel family from rat and human brain. ClC-6 is a 97-kDa protein, and ClC-7 a 89-kDa protein roughly 45% identical with ClC-6. Together they define a new branch of this gene family. Both genes are very broadly expressed, e.g. in brain, testes, muscle and kidney. In mouse embryos, both genes are expressed as early as day 7. While the human gene for ClC-6 is located on human chromosome 1p36 and shares this region with hClC-Ka and hClC-Kb, ClC-7 is on 16p13. ClC-6 has a highly conserved glycosylation site between transmembrane domains D8 and D9, while ClC-7 is the only known eukaryotic ClC protein which lacks this site. Hydropathy analysis indicates that domain D4 cannot serve as a transmembrane domain. Both ClC-6 and ClC-7 cannot be expressed as chloride channels in Xenopus oocytes, either singly or in combination.

Amino Acid Sequence↗

Coat protein gene sequence of an Austrian isolate of grapevine fanleaf virus.

The nucleotide sequence of the coat protein cistron of an Austrian isolate of grapevine fanleaf virus (GFLV-FC) from Vitis vinifera cv. French Colombard was determined. It shows small differences at the RNA level as well as at the protein level compared to the sequences of already published grapevine fanleaf virus strains. The differences may be a result of the natural variation among virus populations or a consequence of selection in a special host plant. As the virus RNA sequence reported here was isolated directly from its natural woody host by an immunocapture-PCR technique, passage of the virus through a herbaceous host could be avoided and a possible bias introduced by a different host environment was excluded. The sequence similarity of known GFLV coat protein cistrons to the sequence described is as high as among the other strains.

Amino Acid Sequence↗

Single dose pharmacokinetics of (S)-atenolol administered orally as a single enantiomer formulation and as a racemic mixture (Tenormin).

The purpose of this study was to describe the pharmacokinetics of and heart rate and blood pressure responses to (S)-atenolol (SATN) and (R)-atenolol (RATN) after oral administration of (S)-atenolol and (R,S)-atenolol (Tenormin) in man. Eight male subjects were given single oral doses of 50 mg of SATN as a single enantiomer formulation (SEF) and 100 mg of Tenormin (TMN) using a randomized, double-blind, 2-period, complete crossover study design. Subjects performed exercise tolerance tests (Bruce Protocol) before and 2, 4, 6, 8, 12, and 24 h after drug administration. Plasma samples were obtained 2 min before and 30 min, 1, 1.5, 2, 3, 4, 5, 6, 8, 10, 12, 16, and 24 h after dosing. Urine was collected for the first 48 h after dosing. Plasma and urine samples were analyzed for SATN and RATN by an enantioselective HPLC method. SEF and Tenormin attenuate exercise-induced increases in heart rate and systolic blood pressure. Mean changes in exercise heart rates 4 h after dosing were -38 +/- 3 bpm and -37 +/- 3 bpm for SEF and TMN, respectively, P = 0.792. Mean changes in exercise systolic blood pressure were -42 +/- 12 mm Hg and -55 +/- 14 mm Hg for SEF and TMN, respectively, P = 0.484. Mean area under the plasma level time curve (AUC0-24) and mean Cmax for SATN for SEF were significantly lower than for SATN after TMN.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

[High altitude lung edema in the East Alps].

We present a case of high-altitude pulmonary edema in a trained skier who undertook a three-day tour in the eastern Alps. This clinical condition can occur in hitherto healthy, predisposed persons at a critical altitude of 2500-3000 m and has to be differentiated above all from pneumonia. Extreme physical exertion, especially at low temperatures, the intake of alcohol or hypnotics and, presumably, preceding infection of the respiratory tract favor the occurrence of high-altitude pulmonary edema. The history of having spent the night at or above the critical altitude and the spontaneous restitutio ad integrum with rapid clinical recovery and dissolution of the radiographic pulmonary infiltrates within a few days after evacuation to lower altitudes are decisive features in establishing the diagnosis. Without therapy or evacuation the mortality rate is 50%. In spite of the low incidence in the eastern Alps, as opposed to the western Alpine countries, the potentially fatal course of high-altitude pulmonary edema should make it mandatory to equip huts and refuges above 3000 m even in the eastern Alps with the first-aid medicament nifedipine and therapeutic oxygen.

Adult↗

Postnatal development of central nervous alpha 2-adrenergic binding sites: an in vitro autoradiography study in the tree shrew.

The postnatal development of the alpha 2-adrenoceptor pattern was investigated by in vitro receptor autoradiography with the antagonist [3H]rauwolscine in the brains of tree shrews (Tupaia belangeri). At birth, high numbers of [3H]rauwolscine binding sites are diffusely distributed in the whole brain with exception of the neocortex which is very weakly labeled at this time. While the number of [3H]rauwolscine binding sites in the cerebellum decreases to low levels during the first three postnatal weeks, several brain regions show a significant increase in binding sites which are progressively concentrated in distinct nuclei. In the medulla oblongata, the diffuse labeling pattern changes so that binding sites become centralized in the dorsomedial nuclei. In the pons, similar changes can be observed with a moderate labeling of the locus coeruleus on postnatal day 10 and a strong labeling in the adult. In the thalamus, a transient appearance of high numbers of [3H]rauwolscine binding sites can be observed during the second and third postnatal week in specific nuclei. In the preoptic area and hypothalamus, there are only minor postnatal changes but the numbers of [3H]rauwolscine binding sites decrease between postnatal day 5 and adulthood. The high number of binding sites in the limbic system does not significantly change after birth. In the neocortex and the superior colliculus, the [3H]rauwolscine labeling pattern which is characteristic for the adult is achieved not before the third postnatal week. Competition experiments demonstrate that [3H]RAUW binds with high affinity to alpha 2-adrenoceptors in the postnatal as well as in the adult brain. Therefore, this study demonstrates region specific developmental profiles of the pattern of alpha 2-adrenoceptors in the postnatal tree shrew brain.

Aging↗

The visual system and levels of perception: properties of neuromental organization.

To see whether the mental and the neural have common attributes that could resolve some of the traditional dichotomies, we review neuroscientific data on the visual system. The results show that neuronal and perceptual function share a parallel and hierarchical architecture which is manifest not only in the anatomy and physiology of the visual system, but also in normal perception and in the deficits caused by lesions in different parts of the system. Based on the description of parallel hierarchical levels of active information processing in the visual brain, we suggest a concept of dissociable levels of perception, advocating that the phenomenal perception and recognition is realized in the functional integrity of a network of reciprocal cortico-cortical connections. The properties shared by neuronal and perceptional functions provide a basis for a neuromental monism in which both functions are attributed a causal role.

Cerebral Cortex↗

Acceptance of direct physician access to a computer-based patient record in a managed care setting.

Kaiser Permanente Mid-Atlantic States has developed a fully integrated outpatient information system which currently runs on an IBM ES9000 on a VM platform written in MUMPS. The applications include Lab, Radiology, Transcription, Appointments. Pharmacy, Encounter tracking, Hospitalizations, Referrals, Phone Advice, Pap tracking, Problem list, Immunization tracking, and Patient demographics. They are department specific and require input and output from a dumb terminal. We have developed a physician's work station to access this information using PC compatible computers running Microsoft Windows and a custom Microsoft Visual Basic 2.0 environment which draws from these 14 applications giving the physician a comprehensive view of all electronic medical records. Through rapid prototyping, voluntary participation, formal training and gradual implementation we have created an enthusiastic response. 95% of our physician PC users access the system each month. The use ranges from 0.2 to 3.0 screens of data viewed per patient visit. This response continues to drive the process toward still greater user acceptance and further practice enhancement.

Ambulatory Care Information Systems↗