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Biomedical subjects

K Witte

Publications and source records attributed to K Witte.

At least 37 records · Page 2Linked to original sources
PubMed2001

Monolayers of derivatized poly(L-lysine)-grafted poly(ethylene glycol) on metal oxides as a class of biomolecular interfaces.

We report on the design and characterization of a class of biomolecular interfaces based on derivatized poly(l-lysine)-grafted poly(ethylene glycol) copolymers adsorbed on negatively charged surfaces. As a model system, we synthesized biotin-derivatized poly(l-lysine)-grafted poly(ethylene glycol) copolymers, PLL-g-[(PEGm)((1-x)) (PEG-biotin)(x)], where x varies from 0 to 1. Monolayers were produced on titanium dioxide substrates and characterized by x-ray photoelectron spectroscopy. The specific biorecognition properties of these biotinylated surfaces were investigated with the use of radiolabeled streptavidin alone and within complex protein mixtures. The PLL-g-PEG-biotin monolayers specifically capture streptavidin, even from a complex protein mixture, while still preventing nonspecific adsorption of other proteins. This streptavidin layer can subsequently capture biotinylated proteins. Finally, with the use of microfluidic networks and protein arraying, we demonstrate the potential of this class of biomolecular interfaces for applications based on protein patterning.

Binding Sites↗
PubMed2001

Time-resolved x-ray K-shell spectra from high density plasmas generated by ultrashort laser pulses.

We present time-resolved x-ray spectra of C, F, Na, and Al, generated by focusing ultrashort frequency doubled Ti:sapphire laser pulses on solid plane targets. Using a high resolution x-ray streak camera in combination with a laser triggered accumulation system, we achieved a time resolution of 1.7 ps when adding the x-ray signal of many thousands of laser shots. K-shell resonance line emission with a duration in the range of 2-4 ps is observed. Ly alpha emission is generally observed to be faster than He alpha emission and the x-ray pulse duration is observed to decrease with increasing atomic number. A hydrodynamic code in combination with an atomic kinetics code is applied for simulation of time-resolved plasma emission, showing good agreement with experimental data.

Journal Article↗
PubMed2001

Phage-display evolution of tyrosine kinases with altered nucleotide specificity.

The problem of identifying downstream targets of kinase phosphorylation remains a challenge despite technological advances in genomics and proteomics. A recent approach involves the generation of kinase mutants that can uniquely use "orthogonal" ATP analogs to phosphorylate substrates in vivo. Using structure-based design, mutants of several protein kinase superfamily members have been found; robust and general methods are needed, however, for altering the nucleotide specificity of the remaining kinases in the genome. Here we demonstrate the application of a new phage display technique for direct functional selection to the identification of a tyrosine kinase mutant with the ability to use N6-benzyl-ATP. Our method produces, in five rounds of selection, a mutant identical to the best orthogonal Src kinase found to date. In addition, we isolate from a larger library of kinase mutants a promiscuous clone capable of using many different ATP analogs. This approach to engineering orthogonal kinases, combined with others, will facilitate the mapping of phosphorylation targets of any kinase in the genome.

Adenosine Triphosphate↗
PubMed2001

Normalisation of blood pressure in hypertensive TGR(mREN2)27 rats by amlodipine vs. enalapril: effects on cardiac hypertrophy and signal transduction pathways.

It is still a controversial issue whether different classes of antihypertensive drugs are equally effective in the regression of cardiac hypertrophy and associated complications. The present study compared the effects of prolonged treatment with the Ca2+-channel blocker amlodipine and the ACE inhibitor enalapril, respectively, in TGR(mREN2)27 rats (TGR), an animal model of renin-dependent hypertension. TGR were divided into three groups and received either amlodipine, enalapril or drinking water without addition, Sprague-Dawley rats (SPRD) served as normotensive control group. Cardiovascular parameters were monitored by radiotelemetry, and drug doses were titrated until 24-h blood pressure was reduced to approximately 140/90 mmHg in both active treatment groups. After 8 weeks of treatment left ventricular (LV) hypertrophy was completely reversed in both treatment groups despite a tenfold increase in plasma angiotensin II in amlodipine-treated TGR. In untreated TGR LV catecholamines were depleted, and beta1-adrenergic stimulation of adenylyl cyclase was blunted. Treatment of TGR with enalapril prevented both the depletion of tissue catecholamines and the desensitisation of LV beta1-adrenoceptors. Amlodipine had no effect on cardiac adrenergic signal transduction. Basal activity of LV soluble guanylyl cyclase was not different between TGR and SPRD, but its sensitivity to stimulation by nitric oxide was slightly reduced in TGR. Treatment had no effect on basal and stimulated guanylyl cyclase activity. The present study in an animal model of renin-dependent hypertension suggests that blood pressure reduction per se is sufficient for a regression of cardiac hypertrophy. However, beta-adrenergic desensitisation was prevented only in the enalapril-treated group, supporting a blood pressure-independent contribution of the renin-angiotensin system to the regulation of beta-adrenergic signal transduction.

Adenylyl Cyclases↗
PubMed2001

Sickness behaviour, an animal equivalent to human quality of life, is improved in septic rats by G-CSF and antibiotic prophylaxis.

BACKGROUND AND AIM: New biological response modifiers are usually tested in reductionistic, pharmacological animal models by the determination of mechanistic endpoints (mortality rate, cellular/physiological parameters). In the meantime, quality of life had become an important endpoint in clinical trials but adequate animal experiments are very rare. The aim of this study was to demonstrate alterations in the behavioural response of septic rats due to a prophylaxis with cytokine (G-CSF) plus antibiotics. METHODS: Sickness behaviour (locomotor activity, circadian rhythms of blood pressure, heart rate and temperature) was determined by the use of radio telemetry. Complex animal experiments in rats were performed including anaesthesia, antibiotic and G-CSF prophylaxis, volume substitution, laparotomy, contamination and infection with human faecal suspension and postoperative analgesia. RESULTS: Prior to infection, rats showed circadian rhythm in locomotor activity, blood pressure, heart rate and temperature. Sham operation did not alter these parameters significantly. Immediately after abdominal contamination and infection, locomotor activity was strongly reduced and circadian rhythm was lost in all parameters. Body temperature showed a continuous rise, peaking 38 h after infection. Untreated animals died in 63% (8/14) of cases. Antibiotic prophylaxis blunted the febrile response and markedly reduced mortality to 20% (2/10) or 0% (0/10) using G-CSF plus antibiotics. Blood pressure and heart rate were increased in parallel with the rise in temperature. These early physiological changes were not prevented by prophylaxis, but normal behaviour was restored faster with G-CSF plus antibiotic prophylaxis. CONCLUSIONS: In septic rats, sickness behaviour (locomotor activity) is significantly improved in parallel to the mortality rate by a prophylaxis with G-CSF plus antibiotics. Sickness behaviour can be considered as an equivalent to human quality of life.

Analysis of Variance↗
PubMed2001

Altered melatonin production in TGR(mREN2)27 rats: on the regulation by adrenergic agonists, antagonists and angiotensin II in cultured pinealocytes.

Transgenic TGR(mREN2)27 rats (TGR), carrying an additional mouse renin gene, are characterized by severe hypertension, an inverse circadian blood pressure profile, a blunted response to photic entrainment signals, and an increased nocturnal production of the pineal hormone melatonin. In order to evaluate the contribution of the over-expressed renin-angiotensin system to the function of the pineal gland in TGR, we studied the adrenergic and angiotensin II (Ang II)-mediated regulation of melatonin synthesis using dispersed pinealocytes from TGR and from Sprague-Dawley control rats (SDR). Isoproterenol was more effective in stimulating melatonin release in pinealocytes from TGR than from SDR, whereas the maximum effect of norepinephrine (NE) stimulation did not differ between the strains. Prazosin reduced the NE-mediated melatonin release only in SDR but not in TGR pinealocytes. Competition experiments with (+/-)-, (+)-, (-)-propranolol and (+/-)-atenolol revealed one homogeneous population of beta1-adrenoceptors. Ang II had no significant effect on basal or isoproterenol-induced melatonin release in either strain. In conclusion, TGR pinealocytes were more sensitive to beta-adrenergic stimulation than SDR pinealocytes, but lacked the alpha1-adrenergic potentiation of beta-adrenergic induced melatonin release. The renin-angiotensin system was not directly involved in the regulation of melatonin synthesis by rat pinealocytes in vitro.

Adrenergic Agonists↗
PubMed2001

Reduced baroreflex sensitivity and blunted endogenous nitric oxide synthesis precede the development of hypertension in TGR(mREN2)27 rats.

Transgenic TGR(mREN2)27 (TGR) rats are an animal model of fulminant hypertension characterized by an inverse circadian blood pressure profile. The present study addressed the contribution of nitric oxide (NO) synthesis and baroreflex function to hypertension and the inverse blood pressure pattern. NO synthesis was measured at four different times of day indirectly by excretion of NO metabolites (NOx: NO2- and NO3-) in the urine of 5- and 11-week-old TGR and Sprague-Dawley (SPRD) controls. Blood pressure, heart rate, and motor activity were recorded in age-matched rats of both strains using an implantable telemetry system. Beat-to-beat recording of blood pressure and pulse interval was performed hourly in 6-week-old animals over 24 h. From these data, baroreflex sensitivity (BRS) was calculated by linear regression of spontaneous fluctuations of blood pressure and corresponding changes of pulse interval. Baroreflex sensitivity was lower in pre-hypertensive TGR rats than in SPRD rats, and the reduction was restricted to the daily resting period. In both strains, NOx excretion showed circadian rhythmicity, with peak values during the activity period at night. Interestingly, excretion of NOx was reduced during the resting period in 5-week-old TGR rats prior to the development of hypertension. Impairment of NO synthesis and baroreflex function precede the development of hypertension in TGR rats. The reduction of both parameters was restricted to the resting period and, therefore, could be involved in the development of the inverse circadian blood pressure profile of TGR rats.

Animals↗
PubMed2001

Influence of circadian time and age on glomerular angiotensin II receptors in normotensive Sprague-Dawley and transgenic hypertensive TGR(mREN2)27 rats.

In male heterozygous transgenic hypertensive rats, TGR(mREN2)27 (TGR), exhibiting an inverse blood pressure profile and in normotensive Sprague-Dawley (SPRD) controls, the density and affinity of angiotensin II receptors were determined at six circadian times in glomeruli of animals 11 weeks old kept under light-dark 12h:12 (LD 12:12) conditions. Angiotensin II receptors were also studied in rats 18-20 weeks old of both strains at 2h after light onset. As a measure of renal excretory functions, diuresis, creatinine, and protein excretion were monitored using metabolic cages. The expression of angiotensin II receptor mRNA was determined in renal arteries 2h-4h after light onset. The following results were obtained: (1) Renal excretory functions showed significant daily variation, with higher excretion rates in the dark span in both TGR and SPRD rats. (2) No circadian phase dependency was found in the glomerular angiotensin II receptors in both rat strains. However, receptor density was significantly lower in TGR than in SPRD rats. In both strains, receptor number increased with aging. (3) In renal arteries, the angiotensin II receptor mRNA of the main receptor subtype AT1A was neither strain nor age dependent, AT1B- and AT2-receptor mRNAs were significantly lower in TGR than SPRD rats. In conclusion, the results demonstrate that the overactive renin-angiotensin system in TGR rats led to a down-regulation of glomerular angiotensin II receptors that was not accompanied by a down-regulation of the mRNA of the dominant AT1A- receptor subtype. Circadian short-term variations in blood pressure in both TGR and SPRD rats are not reflected by daily variation in angiotensin II receptor density of renal glomeruli or by variation in receptor expression in renal vascular tissue.

Age Factors↗
PubMed2001

Cardiovascular regulation in TGR(mREN2)27 rats: 24h variation in plasma catecholamines, angiotensin peptides, and telemetric heart rate variability.

Dysfunction of the sympathetic nervous system might play an important role in disturbed 24h blood pressure regulation in transgenic hypertensive TGR (mREN2)27 (TGR) rats. Our study was performed to determine possible differences in activity of the sympathetic nervous system in TGR rats in comparison to their normotensive Sprague-Dawley (SPRD) controls; we measured plasma catecholamine and angiotensin concentrations throughout 24h under synchronized light-dark 12h:12H (LD 12:12) conditions. In the TGR rat strain, rhythms of plasma catecholamines were blunted, and the concentrations were significantly decreased. In addition, TGR rats showed increased plasma angiotensin I and II concentrations without any significant rhythm. An impaired autonomic regulation was confirmed by monitoring heart rate variability in TGR rats. Data showed that the TGR rat strain is characterized by a reduction in plasma catecholamines and an increase in angiotensin peptides. At present, it is not clear whether the reduction in catecholamines represents a decrease in sympathetic tone mediated by baroreflex activation or an increased catecholamine turnover induced by elevated angiotensin II. However, the blunted, but normally phased, rhythms in plasma catecholamines in TGR rats make it unlikely that the sympathetic nervous system is mainly responsible for the inverse circadian blood pressure rhythm in the transgenic strain.

Angiotensin I↗
PubMed2001

Circadian pacemaker function and entrainment during maturation of transgenic hypertensive TGR(mREN2)27 and Sprague-Dawley rats.

TGR(mREN2)27 (TGR) transgenic rats develop hypertension due to the mouse mRen-2 gene inserted in their genome. At 5 weeks of age, the blood pressure of TGR rats starts rising, until a maximum is reached at 10 weeks of age. Adult TGR rats show peak values of blood pressure (BP) during the light phase, while heart rate (HR) and motor activity (MA) peak at night. In the present experiment, we evaluated the evolution of circadian rhythms in motor activity, heart rate, and blood pressure of TGR and Sprague-Dawley (SD) rats under 12h light-dark cycles (LD 12:12). Results confirmed that the blood pressure of TGR rats starts to increase at 5 weeks of age, reaching a plateau by the 11th week. Parallel to the increase in blood pressure levels, there was a decrease in the period length of the blood pressure rhythm, a delay in the onset of the alpha phase of the blood pressure rhythm with respect to that of motor activity and heart rate, and a decrease in heart rate levels. In all of the variables studied, the alpha phase of SD rats always started before darkness, whereas that of TGR rats started after lights off. In general, heart rate and motor activity levels of TGR rats were higher than those of SD rats. The amplitude of the circadian rhythms studied was greater in TGR rats than in SD rats. The present results suggest that the different evolution of circadian rhythms in TGR and SD rats might be due to differences in the functioning of the entrainment pathway or the circadian clock itself, which can be detected in young rats and that are probably caused by the expression of the mouse transgene.

Aging↗
PubMed2001

Effect of short light-dark cycles on young and adult TGR(mREN2)27 rats.

Animals placed under short light-dark (LD) cycles show a dissociation of their circadian rhythms. However, this effect has only been studied in Wistar rats and with the motor activity (MA) rhythm. Thus, in the present experiment, we studied in TGR(mREN2)27 (TGR) rats, a strain of hypertensive rats, the effect of a short LD cycle on the circadian rhythms of MA, heart rate (HR), and blood pressure (BP). Our aim was (1) to investigate whether the exposure of TGR rats to a short LD cycle induced a dissociation of their circadian rhythms, (2) to study the effect of short LD cycles on the development of the circadian rhythms of TGR rats, and (3) to compare the effect of short LD cycles on young and adult TGR rats. One group of TGR rats was maintained under LD cycles of 22h periods (group G22). The progress in time of their rhythms was compared to that of TGR rats of the same age that had been kept under LD cycles of 24h periods (group G24). For the third point, the rhythms of a group of 5-week-old TGR rats kept under LD 22h cycles (young rats) were compared to those of a group of 11-week-old TGR rats (adult rats). Results showed that there is a dissociation of the circadian rhythms of all the variables monitored in TGR rats maintained under LD 22h cycles, independent of age. We have also found that group G22 showed a higher increase in BP with age and a higher mortality due to malignant hypertension compared to group G24. Finally, it seems that it is harder for young rats to entrain to short LD cycles than for adult rats, and young rats have a higher mortality due to malignant hypertension than adult rats. In conclusion, we demonstrated that short LD cycles produce a dissociation in the HR, BP, and MA circadian rhythms. The results of this experiment, compared to those previously obtained in Wistar rats, suggest that the light perception, the responses of the circadian system to light, or both are altered in the TGR rats.

Aging↗
PubMed2001

Day-night variation in the in vitro contractility of aorta and mesenteric and renal arteries in transgenic hypertensive rats.

TGR(mREN2)27 (TGR) rats develop severe hypertension and an inverted circadian blood pressure profile with peak blood pressure in the day-time rest phase. The present study investigated the in vitro responsiveness of different arteries of TGR rats during day and night. Twelve-week-old TGR rats and normotensive Sprague-Dawley (SPRD) controls, synchronized to 12h light, 12h dark (LD 12:12) (light 07:00-19:00), were killed at 09:00 (during rest) and 21:00 (during activity), and endothelium-dependent relaxation by acetylcholine and vascular contraction by angiotensin II were studied by measuring isometric force in ring segments of abdominal aorta and mesenteric and renal arteries. In SPRD rats, consistent day-night variation was found, with greater responses to angiotensin II during the daytime rest span. In TGR rats, biological time-dependent differences were found in the renal vasculature, but not in the aorta and mesenteric artery. Relaxation of SPRD rat aorta and mesenteric artery by acetylcholine was greater at 09:00, whereas in TGR rats, day-night variation was absent (mesenteric artery) or inverted (aorta). In conclusion, based on the study of two time points, day-night variation in vascular contractility of aorta and mesenteric artery is blunted in TGR rats, whereas renal artery segments showed an unchanged day-night pattern compared to SPRD controls.

Acetylcholine↗
PubMed2000

Loss of 24 h rhythm and light-induced c-fos mRNA expression in the suprachiasmatic nucleus of the transgenic hypertensive TGR(mRen2)27 rat and effects on cardiovascular rhythms.

Immediate early genes, especially c-fos, are thought to play an essential role in photic entrainment of circadian rhythms. A special characteristic of the transgenic hypertensive TGR(mRen2)27 rat strain, expressing an additional mouse renin2 gene, is the inverse blood pressure rhythm in relation to those in heart rate and activity resulting in internal desynchronisation of these physiological rhythms. Assessment of c-fos mRNA expression by microdissection and RT-PCR in the suprachiasmatic nucleus showed, that in contrast to normotensive Sprague-Dawley rats the 24 h and circadian rhythm of c-fos mRNA expression in TGR(mRen2)27 rats is abolished. Moreover, light-induced c-fos expression within the nucleus could be found in the normotensive controls, but was absent in transgenic hypertensive rats. The light pulse applied during the subjective night, at CT 14, significantly phase delayed rhythms in blood pressure, heart rate and activity in the normotensive rats by about 2 h, whereas in the transgenic hypertensive animals rhythms in blood pressure and heart rate were unaffected, only activity showed a slight phase shift. In conclusion, these data suggest that the transgene in TGR leads not only to a disturbance of the cardiovascular system but also influences the light entrainment response, which is accompanied by a suppressed c-fos mRNA expression in the suprachiasmatic nucleus.

Animals↗
PubMed2000

Contribution of the renin-angiotensin system to subsensitivity of soluble guanylyl cyclase in TGR(mREN2)27 rats.

Soluble guanylyl cyclase activity and its stimulation by diethylamineNONOate was measured in aortae from hypertensive TGR(mREN2)27 rats (TGR) and Sprague-Dawley controls. Superoxide dismutase was added in vitro to evaluate the contribution of oxidative breakdown of nitric oxide (NO) by superoxide anions. Expression of soluble guanylyl cyclase was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Basal and stimulated soluble guanylyl cyclase activity was significantly reduced in TGR rats, addition of superoxide dismutase had no effect. Expression of soluble guanylyl cyclase subunits was not different between strains. The independent contribution of hypertension and the overactive renin-angiotensin system to soluble guanylyl cyclase subsensitivity was assessed after normalization of TGR's blood pressure by the Ca(2+)-channel blocker amlodipine or the angiotensin converting enzyme-inhibitor enalapril. Soluble guanylyl cyclase activity in TGR was slightly increased by amlodipine and almost completely restored by enalapril. In conclusion, TGR showed desensitized vascular soluble guanylyl cyclase, depending on their overactive renin-angiotensin system.

Aging↗
PubMed2000

Impact of a worker notification program: assessment of attitudinal and behavioral outcomes.

BACKGROUND: Hundreds of worker notification programs are conducted each year to communicate occupational risks to workers. However, few attempts have been made to evaluate their effectiveness and few have described how communication theories are applied in developing notification messages. We developed and assessed the effectiveness of a worker notification program at a beryllium machining plant. METHODS: We compared self-protective attitudinal and behavioral responses among workers in two plants: (1) an intervention plant that received beryllium risk notification and (2) a matched control plant that did not receive notification. RESULTS: Workers receiving notification reported significantly stronger perceptions of threat and efficacy, more positive attitudes toward safety practices, and engaged in more protective behaviors than the workers at the control plant. CONCLUSIONS: This study demonstrates the utility of applying communication theories in the development of notification messages and the results suggest that mass presentations may be just as effective, if not more so, than one-on-one notifications. Am. J. Ind. Med. 37:205-213, 2000. Published 2000 Wiley-Liss, Inc.

Adult↗
PubMed2000

Preventing vehicle crashes with trains at grade crossings: the risk seeker challenge.

A formative evaluation for a communication campaign to decrease rail-automobile accidents was conducted with a survey of 891 randomly selected residents in Michigan, USA. The formative evaluation was theoretically grounded in the extended parallel process model. The results of the study suggest that the majority of respondents engage in safe driving behaviors around railways. However, 10-20% reported extremely risky behaviors such as trying to 'beat the train' (labeled 'risk seekers'). Further analyses revealed that the risk seekers were disproportionately male with strong sensation seeking tendencies to engage in new and novel experiences and to avoid boredom. The results suggest that high sensation seeking tendencies cause one to experience greater frustration and exhibit greater judgment distortions around rail crossings, which in turn, cause one to try and beat the train. Specific recommendations are given for campaign developers and limitations to the study are addressed.

Accidents, Traffic↗
PubMed2000

Experimental heart failure in rats: effects on cardiovascular circadian rhythms and on myocardial beta-adrenergic signaling.

OBJECTIVES: Patients with chronic heart failure frequently show blunted circadian blood pressure profiles. The mechanisms involved in the loss of physiological day-night variation are still unclear, but a continuously active sympathetic nervous system could play a role. The present study evaluated long-term consequences of rat heart failure on cardiovascular circadian patterns in vivo, and on density and function of cardiac beta-adrenoceptor subtypes in vitro, as a marker of cardiac adrenergic drive. METHODS: Heart failure in rats was induced by coronary artery ligation leading to infarct sizes of >30% of left ventricular circumference. Blood pressure and heart rate were monitored for 10 weeks after infarction using radiotelemetry. Density and function of cardiac beta(1) and beta(2)-adrenoceptors were measured by radioligand binding and adenylyl cyclase stimulation. RESULTS: During the activity period at night blood pressure and heart rate were lower in rats with heart failure than in sham controls, leading to reduced night-day variation in the heart failure group. Depression of circadian rhythmicity in blood pressure was found over the whole study period, while that in heart rate occurred with a lag-time of several weeks. In failing left ventricles beta-adrenoceptors showed reduced high affinity agonist binding, a shift in the beta(1):beta(2) ratio towards the beta(2)-subtype, and decreased beta(1)-adrenergic stimulation of adenylyl cyclase. In right ventricles no differences were found between failing and control rats. The blunted nocturnal increase in blood pressure and heart rate as well as beta(1)-adrenergic desensitization were correlated with the severity of left ventricular dysfunction. CONCLUSIONS: Heart failure in rats leads to disturbed circadian patterns in blood pressure and heart rate, and to desensitization of cardiac beta(1)-adrenoceptors, indicating chronic sympathetic overactivity.

5'-Nucleotidase↗