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Biomedical subjects

C Koch

Publications and source records attributed to C Koch.

At least 271 records · Page 15Linked to original sources

Antibody response in rabbits infected with Rochalimaea henselae, Rochalimaea quintana and Afipia felis.

Antibody responses in three pairs of rabbits inoculated with live Rochalimaea henselae, Rochalimaea quintana and Afipia felis were studied by enzyme immunoassay with whole-cell and bacterial sonicates as antigen. No differences in measured antibody responses were found with the two types of antigen preparation. Two rabbits did not respond with antibody production. In the remaining rabbits there was a low-titred antibody response that showed no significant cross-reaction with related bacteria. After rechallenge the antibody response rose significantly and there was significant cross-reaction with related bacteria. The antigens involved in the antibody response were examined by crossed immunoelectrophoresis. After the initial inoculation 5-7 precipitin lines of the reference diagrams were deflected, including lines which cross-reacted with antigens found in related bacterial species. After reinoculation several more precipitin lines were deflected, including additional lines cross-reacting with antigens present in related bacteria and common bacterial antigens.

Animals↗

Linearized models of calcium dynamics: formal equivalence to the cable equation.

The dynamics of calcium and other diffusible second messengers play an important role in intracellular signaling. We show here the conditions under which nonlinear equations governing the diffusion, extrusion, and buffering of calcium can be linearized. Because the resulting partial differential equation is formally identical to the one-dimensional cable equation, quantities analogous to the input resistance, space constant, and time constant--familiar from the study of passive electrical propagation--can be defined. Using simulated calcium dynamics in an infinite cable and in a dendritic spine as examples, we bound the errors due to the linearization, and show that parameter uncertainty is so large that most nonlinearities can usually be ignored: robust phenomena in the nonlinear model are also present in the linear model.

Animals↗

Power spectrum analysis of bursting cells in area MT in the behaving monkey.

It is widely held that visual cortical neurons encode information primarily in their mean firing rates. Some proposals, however, emphasize the information potentially available in the temporal structure of spike trains (Optican and Richmond, 1987; Bialek et al., 1991), in particular with respect to stimulus-related synchronized oscillations in the 30-70 Hz range (Eckhorn et al., 1988; Gray et al., 1989; Kreiter and Singer, 1992) as well as via bursting cells (Cattaneo et al., 1981a; Bonds, 1992). We investigate the temporal fine structure of spike trains recorded in extrastriate area MT of the trained macaque monkey, a region that plays a major role in processing motion information. The data were recorded while the monkey performed a near-threshold direction discrimination task so that both physiological and psychophysical data could be obtained on the same set of trials (Britten et al., 1992). We identify bursting cells and quantify their properties, in particular in relation to the behavior of the animal. We compute the power spectrum and the distribution of interspike intervals (ISIs) associated with individual spike trains from 212 cells, averaging these quantities across similar trials. (1) About 33% of the cells have a relatively flat power spectrum with a dip at low temporal frequencies. We analytically derive the power spectrum of a Poisson process with refractory period and show that it matches the observed spectrum of these cells. (2) About 62% of the cells have a peak in the 20-60 Hz frequency band. In about 10% of all cells, this peak is at least twice the height of its base. The presence of such a peak strongly correlates with a tendency of the cell to respond in bursts, that is, two to four spikes within 2-8 msec. For 93% of cells, the shape of the power spectrum did not change dramatically with stimulus conditions. (3) Both the ISI distribution and the power spectrum of the vast majority of bursting cells are compatible with the notion that these cells fire Poisson-distributed bursts, with a burst-related refractory period. Thus, for our stimulus conditions, no explicitly oscillating neuronal process is required to yield a peak in the power spectrum. (4) We found no statistically significant relationship between the peak in the power spectrum and psychophysical measures of the monkeys' performance on the direction discrimination task.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Structural analysis of chicken factor B-like protease and comparison with mammalian complement proteins factor B and C2.

Chicken complement factor B-like protease is a glycoprotein of 95 kDa. Activation of chicken serum complement with inulin cleaved the B-like protease into an N-terminal Ba fragment of 37 kDa and a C-terminal Bb fragment of 60 kDa. The whole protein and the two fragments were purified by affinity chromatography using mAb to chicken Ba or Bb followed by ion exchange chromatography. Amino acid sequencing showed that chicken B-like protease was cleaved at a site homologous to that cleaved in mammalian complement components B and C2 on activation. Limited tryptic digestion of the B-like protease generated fragments similar to Ba and Bb. More than 200 residues of the Ba sequence and two N-linked glycosylation sites were established by amino acid sequencing of peptides derived by digestion with four proteases. Comparison of human and mouse C2 and B sequences indicated a slower evolutionary rate for B (85% sequence identity) than for C2 (74% sequence identity). Comparison of chicken Ba to human and mouse C2b and Ba showed 42 to 45% sequence identity with respect to C2b fragments, and 46 to 49% sequence identity with respect to Ba fragments. Taking the slower evolutionary rate of factor B into account, chicken factor B-like protease seems to be equally related to mammalian complement components B and C2, and the B-like protease most likely represents the present-day descendant of a common ancestral protein for mammalian B and C2. This conclusion is in agreement with the requirement for the B-like protease in both classical and alternative activation pathways for chicken complement, and with the apparent lack of a chicken serum protein with exclusive C2 activity.

Amino Acid Sequence↗

A role for the transcription factors Mbp1 and Swi4 in progression from G1 to S phase.

In budding yeast genes that encode G1 cyclins and proteins involved in DNA synthesis are transcriptionally activated in late G1. A transcription factor, called SBF, is composed of Swi4 and Swi6 proteins and activates transcription of G1 cyclin genes. A different, but related, complex called MBF binds to MCB elements (Mlu I cell cycle box) found in the promoter of most DNA synthesis genes. MBF contains Swi6 and a 120-kilodalton protein (p120). MBF was purified and the gene encoding p120 (termed MBP1) was cloned. A deletion of MBP1 was not lethal but led to deregulated expression of DNA synthesis genes, indicating a direct regulatory role for MBF in MCB-driven transcription. Mbp1 is related to Swi4. Strains deleted for both MBP1 and SWI4 were inviable, demonstrating that transcriptional activation by MBF and SBF has an important role in the transition from G1 to S phase.

Amino Acid Sequence↗

Monoclonal antibodies against brain acetylcholinesterases which recognize the subunits bearing the hydrophobic anchor.

Monoclonal antibodies were raised against amphiphilic detergent-soluble (DS) acetylcholinesterase (AChE) from human brain caudate nucleus. Three mAb, 132-4 (IgG1), 132-5 (IgG1) and 132-6 (IgG3), specific for brain DS-AChE were selected and subcloned. These mAb reacted with native as well as heat-denatured and SDS-denatured DS-AChE, indicating that the epitopes to which mAb bound are continuous determinants. The mAb cross-reacted with DS-AChE from bovine and mouse brain and with brain DS-AChE from river trout (Salmo trutta forma fario) and lake trout (Salmo trutta forma lacustris). No cross-reaction was detected with the following antigens: salt-soluble (SS) AChE from bovine brain, glycophospholipid-anchored AChE from human and bovine erythrocytes, DS-butyrylcholinesterase and SS-butyrylcholinesterase (BtChE) from the brains of human and bovine, DS-BtChE from chicken and BtChE from human serum. Deglycosylation of brain DS-AChE with N-glycosidase F did not abolish the binding of mAb to DS-AChE. After reduction of brain DS-AChE by dithiothreitol, the mAb no longer reacted with the antigen, indicating that a disulfide bridge is important for the epitope. Monomerization of brain DS-AChE by trypsin and limited proteinase K treatment also abolished the binding of mAb to DS-AChE. Sucrose-density-gradient centrifugation showed that mAb reacted only with native tetrameric forms, but not with dimeric and monomeric forms. Western blot, after SDS/PAGE under non-reducing conditions, showed that mAb reacted with those subunits carrying the hydrophobic anchor (i.e. tetramers, trimers and heavy dimers) but not with those devoid of it (light dimers or monomers). Since mAb 132-4, 132-5 and 132-6 recognized DS-AChE from fish up to mammalian brain in the evolutionary tree, it is concluded that the epitope to which these mAb bind, is conserved in nature.

Acetylcholinesterase↗

Transcription factors important for starting the cell cycle in yeast.

Unlike early embryonic cleavage divisions in certain animals, cell-cycle progression in yeast and probably also in all metazoan somatic cells requires the periodic transcriptional activation of certain key genes. Thus far, the only clear examples are genes that encode a class of unstable 'cyclin' proteins, which bind and activate the cdc2/Cdc28 protein kinase: the G1-specific cyclins encoded by CLN1 and CLN2, a B-type cyclin implicated in DNA replication encoded by CLB5; and four B-type cyclins involved in mitosis encoded by CLB1, 2, 3, 4. CLN1, CLN2, and CLB5 are transcribed in late G1, as cells undergo Start. A transcription factor composed of Swi4 and Swi6 proteins (called SBF) activates CLN1 and CLN2 transcription via a positive feedback loop in which Cln proteins activate their own transcription. A different but related transcription factor called MBF seems responsible for the late G1-specific transcription of most DNA replication genes including CLB5. We have purified MBF and shown that it contains Swi6 and a 110-120 kDa protein distinct from Swi4 (p120) that contacts DNA. Thus, we propose that SBF and MBF share a common regulatory subunit (Swi6) but recognize their promoter elements via distinct DNA binding subunits.

Base Sequence↗

Good vibes.

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Brain↗

Anti-hirudin monoclonal antibodies directed toward discontinuous epitopes of the hirudin amino-terminal and epitopes involving the carboxy-terminal hirudin amino acids.

A panel of eight monoclonal antibodies (MAbs) was obtained against recombinant hirudin variant 2 (rHV2). Specificities of the eight MAbs indicate that four of them recognize C-terminal amino acid residues (Group A) and four are directed against discontinuous epitopes and recognize a determinant (or determinants) within the 43 N-terminal residues (Group B). Using these antibodies recombinant hirudins missing one or more C-terminal amino acids can be distinguished from molecules with an intact C-terminus either in enzyme immunoassays (EIAs) or by immunoaffinity chromatography. A sandwich EIA using the combination of two antibodies, one from each group, can quantitate both recombinant hirudin variant 1 (rHV1) and rHV2 with a detection range from 1 to 10 ng/ml in either buffer or plasma. Using only one MAb a competitive antibody capture EIA can quantitate recombinant or natural hirudin variants 1, 2, and 3 with a detection range from 5 to 100 ng/ml for rHV2 with a lysine in position 47 (rHV2K47). None of the antibodies recognizes hirudin after it is complexed to alpha-thrombin. The ability of any one of these anti-rHV2 antibodies to interfere with hirudin binding to alpha-thrombin as measured by inhibition of thrombin's amidolytic activity correlates with the range of MAb affinity constants (KD = 3.5 x 10(-9) to 1 x 10(-6) M). Incubating hirudin with one antibody from Group A (KD = 3.5 x 10(-8) M) and one from Group B (KD = 6.0 x 10(-9) M) completely blocks the ability of hirudin to bind alpha-thrombin. This MAb panel is thus useful for probing the recombinant C-terminal integrity of hirudin, for sensitive free hirudin quantitations, and the combined use of two MAbs has potential applications as an antidote for hirudin in vivo.

Amino Acid Sequence↗

An oscillation-based model for the neuronal basis of attention.

We propose a model for the neuronal implementation of selective visual attention based on the temporal structure of neuronal activity. In particular, we set out to explain the electrophysiological data from areas V4 and IT in monkey cortex of Moran and Desimone [(1985) Science, 229, 782-784] using the "temporal tagging" hypothesis of Crick and Koch [(1990a) Cold Spring Harbor Symposiums in Quantitative Biology, LV, 953-962; (1990b) Seminars in the neurosciences (pp. 1-36)]. Neurons in primary visual cortex respond to visual stimuli with a Poisson distributed spike train with an appropriate, stimulus-dependent mean firing rate. The firing rate of neurons whose receptive fields overlap with the "focus of attention" is modulated with a periodic function in the 40 Hz range, such that their mean firing rate is identical to the mean firing rate of neurons in "non-attended" areas. This modulation is detected by inhibitory interneurons in V4 and is used to suppress the response of V4 cells associated with non-attended visual stimuli. Using very simple single-cell models, we obtain quantitative agreement with Moran and Desimone's (1985) experiments.

Action Potentials↗

Computational approaches to cognition: the bottom-up view.

How can higher level aspects of cognition, such as figure-ground segregation, object recognition, selective focal attention and ultimately even awareness, be implemented at the level of synapses and neurons? A number of theoretical studies emerging out of the connectionist and the computational neuroscience communities are starting to address these issues using neural plausible models.

Animals↗

Pulmonary function in adolescents with childhood asthma.

The aim of this study was to determine the pulmonary function in former and present asthmatics. We examined 77 persons aged 12-24 years, classified into four groups: 1) healthy subjects (controls) (n = 19), 2) former asthmatics (n = 19), 3) present mild asthmatics (n = 20), and 4) present severe asthmatics (n = 19). Although exhibiting no respiratory symptoms, former asthmatics had reduced airflow values measured by FEV1 (median (range) 89.7 (83-99) vs 101.4 (91-110)) and MEF25 (76.5 (68-94) vs 103.0 (97-124)), as compared with controls. Furthermore, former asthmatics had significantly increased PEF variability, as compared with controls, whereas no significant differences were found in static lung parameters, i.e. total lung capacity and residual volume, as compared with controls. In conclusion, former asthmatics, although now exhibiting no respiratory symptoms, were found to have obstructive airflow limitation, increased bronchial responsiveness, and normal lung volumes.

Adolescent↗

Diabetes mellitus in cystic fibrosis: genetic and immunological markers.

Family history, as well as genetic and immunological markers of diabetes mellitus, were studied in cystic fibrosis (CF) patients with and without diabetes mellitus. Positive family history of diabetes mellitus in first-degree relatives was found in only 6 of 210 (3%) CF patients, with no difference between non-diabetic and diabetic patients. The frequency distributions of the HLA types DR3, DR4 and DR3/4, which normally confer susceptibility to insulin-dependent diabetes mellitus and of HLA-DR2, which normally confers resistance to insulin-dependent diabetes mellitus, were not different in non-diabetic CF patients, diabetic CF patients and normal subjects. The genotypic frequencies of tumor necrosis factor-beta and of heat shock protein 70, located within the HLA region on chromosome 6, in CF patients with diabetes were not different from those in patients with insulin-dependent diabetes mellitus, while non-diabetic CF patients and normal subjects shared other patterns. The frequencies of the interleukin-1 beta alleles, located on chromosome 2, were not different in non-diabetic and diabetic CF patients, insulin-dependent diabetic patients and normal subjects. Islet cell cytoplasmic antibodies, measured before, at and after the diagnosis of diabetes in 33 diabetic CF patients and in 32 matched non-diabetic CF patients, were detected in only 2 of 236 (0.8%) serum samples: in a pre-diabetic patient and in a non-diabetic control patient. Birth weights were not different in diabetic and non-diabetic CF patients, arguing against the importance of the intrauterine environment as a determinant in the transmission of diabetes mellitus in CF patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Increased cyclic guanosine monophosphate production and overexpression of atrial natriuretic peptide A-receptor mRNA in spontaneously hypertensive rats.

Atrial natriuretic peptide (ANP) specifically stimulates particulate guanylate cyclase, and cyclic guanosine monophosphate (cGMP) has been recognized as its second messenger. Spontaneously hypertensive rats (SHR) have elevated plasma ANP levels, but manifest an exaggerated natriuretic and diuretic response to exogenous ANP when compared to normotensive strains. In isolated glomeruli, the maximal cGMP response to ANP corresponds to a 12- to 14-fold increase over basal levels in normotensive strains (Wistar 13 +/- 2; Wistar-Kyoto 12 +/- 2; Sprague-Dawley 14 +/- 2) while a maximal 33 +/- 3-fold elevation occurs in SHR (P < 0.001). This hyperresponsiveness of cGMP is reproducible in intact glomeruli from SHR from various commercial sources. Furthermore, this abnormality develops early in life, even before hypertension is clearly established, and persists despite pharmacological modulation of blood pressure, indicating that it is a primary event in hypertension. In vitro studies have revealed a higher particulate guanylate cyclase activity in membranes from glomeruli and other tissues from SHR. This increase is not accounted for by different patterns of ANP binding to its receptor subtypes between normotensive and hypertensive strains, as assessed by competitive displacement with C-ANP102-121, an analog which selectively binds to one ANP receptor subtype. The hyperactivity of particulate guanylate cyclase in SHR and its behavior under basal, ligand (ANP), and detergent-enhanced conditions could be attributed either to increased expression or augmented sensitivity of the enzyme. Radiation-inactivation analysis does not evoke a disturbance in the size of regulatory elements normally repressing enzymatic activity, while the expression of particulate guanylate cyclase gene using mutated standard of A- and B-receptors partial cDNAs, quantified by polymerase chain reaction (PCR) transcript titration assay, manifests a selective increase of one guanylate cyclase subtype. Our data suggest that in hypertension, genetic overexpression of the ANP A-receptor subtype is related to the exaggerated biological response to ANP in this disease.

Affinity Labels↗