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PubMed2007

Study on the interaction between the chiral drug of propranolol and alpha1-acid glycoprotein by fluorescence spectrophotometry.

The interaction between the chiral drug of propranolol (PPL) and alpha1-acid glycoprotein (AGP, orosomucoid) has been first studied by fluorescence spectrophotometry. The fluorescence intensity of PPL increased due to the addition of AGP into PPL. The equation of Scatchard was employed to calculate the association constant and binding site number of the two enantiomers with AGP. The association constant is 2.62 x 10(5)M-1 for R-PPL and 8.57 x 10(5)M-1 for S-PPL and the binding site number is 0.41 for R-PPL and 1.17 for S-PPL at 17 degrees C respectively. The method of thermodynamics was applied to determine the binding type of S-PPL with AGP. The results suggested that the binding type is mainly van der waals force or hydrogen bond. At last the effect of three metal cations on the association constant and the binding site number of S-PPL with AGP was examined.

Orosomucoid↗
PubMedDate not supplied

Evaluation of fixed wavelength fluorescence and synchronous fluorescence spectrophotometry as a biomonitoring tool of environmental contamination.

A field study was carried out to investigate whether polyaromatic hydrocarbon (PAH) contamination in shore crabs (Carcinus maenas) could be measured using fluorescence spectrometry. Crabs were collected as part of an integrated study to assess PAH contamination in Southampton Water, UK. Urine was collected from crabs via a non-destructive technique and analysed using fixed wavelength fluorescence (FF) and synchronous fluorescence spectrophotometry (SFS) in order to determine the presence of PAH metabolites, indicative of PAH exposure. By scanning at wavelength pairs specific to a variety of PAH groups the fluorescence of metabolites was analysed and attributed to the following PAH groups; naphthalenes, pyrenes and benzo[a]pyrene (BAP). Naphthalene-type, pyrene-type and BAP-type metabolites were detected using FF and SFS techniques and two distinct parameters were measured; Peak Intensity and Peak Area. Results showed that crabs obtained near a petroleum refinery, a point source of PAH discharge, exhibited significantly higher PAH metabolites for all PAH groups, than at sites sampled along the estuary (ANOVA, p<0.05). This study highlights the effectiveness of using FF and SFS as a rapid and cost-effective biomonitoring tool for detecting PAH contamination.

Analysis of Variance↗
PubMed1999

The interaction between human and bovine serum albumin and zinc studied by a competitive spectrophotometry.

The binding of zinc to human serum albumin (HSA) and bovine serum albumin (BSA) has been studied by a competitive spectrophotometry. The indictor ligand, 5-Br-PAPS, as well as its metal complexes, ML and ML2, having intense colors with sufficient separate absorption peaks permits the analysis of the interaction between zinc and HSA or BSA. The results show that zinc binds to both albumins in the molar ratio of 1:1 at pH 6-8. The apparent association constants of Zn-HSA and Zn-BSA decrease with decreasing pH. The pH dependence of the constants for both Zn-HSA and Zn-BSA indicates the 1:1 competition between metal ions and hydrogen ions for the binding site. The intrinsic association constants of the metal and the proton are found to be log KZn-HSA = 7.1 +/- 0.2 and log KH-HSA = 8.2 +/- 0.3 for HSA and log KZn-BSA = 7.6 +/- 0.2 and log KH-BSA = 8.2 +/- 0.3 for BSA, respectively.

Animals↗
PubMed1997

A novel near infra-red spectrophotometry system using microprobes: its evaluation and application for monitoring neuronal activity in the visual cortex.

A novel near-infrared (NIR) spectrophotometry system with microprobes of optical transmitter and receivers (550 and 410 microm in diameter, respectively) has been developed. A three-dimensional profile of the signal source estimated in in vitro experiments showed two spindle-shaped regions around the respective probes, suggesting that the signal detected by the present system comes from a relatively restricted region around each probe. Next, we examined how the concentration of oxygen in inspired gas affected the NIR signals in the rat cerebral cortex in vivo. Calculated concentrations of oxyhemoglobin (HbO2) and deoxyhemoglobin (Hb) in the rat cortex changed in mirror-image fashion in response to the change in the O2 concentration in the inspired gas. Finally, NIR responses to visual stimulation were recorded from the striate cortex of conscious adult cats that had been monocularly deprived since the peak of the sensitive period. Corresponding to the results of previous electrophysiological studies, stimulation of the normal eye induced significant NIR signals, whereas that of the deprived eye evoked no response. These results indicate that this new NIR system can be applied to study changes in oxygen metabolism in relatively restricted regions following neuronal activation in the brain.

Animals↗
PubMed2003

Cerebral oxygenation determined by near-infrared spectrophotometry in patients with fulminant hepatic failure.

BACKGROUND/AIMS: In severe cases of acute liver failure (ALF), cerebral hyperperfusion may result in high intracranial pressure and brain damage. The aim of this study was to determine if near-infrared spectrophotometry (NIRS) could detect a raise in cerebral blood flow and oxygenation induced by noradrenaline (NA) infusion. METHODS: In seven ALF patients (five females and two males; median age 49 years (range 20-70)) changes in cerebral concentration of oxy-(deltaHbO(2)) and total-haemoglobin (deltaHbT) were compared to the jugular bulb saturation (SvjO(2)) and cerebral blood flow velocity (Vmean) during NA infusion. RESULTS: Mean arterial pressure increased from 68 (64-86) to 103 (87-118) mmHg and the cerebral perfusion pressure from 61 (53-79) to 95 (74-110) mmHg (P<0.05), while the intracranial pressure (7 (6-15) mmHg) was not significantly changed. In six patients cerebral deltaHbO(2) and deltaHbT increased 2.7 (0.3-9.6) and 2.0 (0.3-14.8) micromol l(-1), respectively, but cerebral oxygenation decreased in one patient. SvjO(2) increased from 68 (55-76) to 74 (64-78) % (P<0.05) concomitant with an increase in Vmean from 47 (34-65) to 68 (50-86) cm s(-1) (P<0.05). deltaHbO(2) covariated with changes in SvjO(2) during NA in all but one patient. CONCLUSIONS: In ALF patients, a change in cerebral perfusion was detected by NIRS. The combination of NIRS and transcranial Doppler sonography may be valuable non-invasive techniques to detect cerebral hyperperfusion before intracranial hypertension becomes manifest.

Adult↗
PubMed2003

Detecting river pollution using fluorescence spectrophotometry: case studies from the Ouseburn, NE England.

Recent advances in fluorescence spectrophotometry enable the analysis of river dissolved organic matter. We investigate the potential of detecting sewage pollution in a small, urbanised catchment. Downstream sampling highlighted a summer maximum in tryptophan fluorescence intensity during low flow. No correlation is observed between ammonia and tryptophan fluorescence intensity. In contrast, two sewage related point-pollution events had both high tryptophan fluorescence intensity and ammonia, suggesting that the summer tryptophan increase does not original from foul sewage. Sewage inputs to the river were therefore monitored at summer base flow. This demonstrated that > 10% of the rivers' discharge is provided by sewerage inputs and that these inputs could be grouped by their fluorescence and ammonia properties: (1) 'clean' storm waters with low ammonia and tryptophan intensity (2) 'grey' waters with high tryptophan intensity and low ammonia concentration, and (3) 'foul' waters with high tryptophan intensity and ammonia concentration. All three types of sewerage input occurred irrespective of flow conditions,suggesting that sewerage cross connections are occurring.

Ammonia↗
PubMed1992

Photocycle of phoborhodopsin from haloalkaliphilic bacterium (Natronobacterium pharaonis) studied by low-temperature spectrophotometry.

Phoborhodopsin (pR) is the fourth retinal pigment of Halobacterium halobium and works as a photoreceptor for the negative phototactic response. A similar pigment was previously found in haloalkaliphilic bacterium (Natronbacterium pharaonis) and also works as the receptor of the negative phototactic response; this pigment is called pharaonis phoborhodopsin (ppR). In this paper, the photocycle of ppR was investigated by means of low-temperature spectrophotometry. The absorption maximum of ppR is located at 498 nm, while that of pR is at 487 nm. The absorption spectra of the two have similar vibrational structures. Irradiation of ppR below -100 degrees C produced a K-like intermediate (ppRK) which was a composite of two components. The original ppR and ppRK were perfectly photoreversible. On warming, ppRK was directly converted to an M-like intermediate without formation of the L-like intermediate. The M-like intermediate was converted to the O-like intermediate at pH 7.2, but the O-like intermediate was not detected at pH 9.0. The O-like intermediate then reverted to the original pigment. On the basis of these findings, the photocycle and the primary photochemical process of ppR are presented.

Archaeal Proteins↗
PubMed2000

Determination of trace selenium in human plasma and hair with ternary inclusion compound-fluorescent spectrophotometry.

A method for determination of selenium in plasma and hair with ternary inclusion compound-fluorescent spectrophotometry has been developed. The determination of selenium in plasma and hair can be performed directly in aqueous solution. Blood and hair samples were destroyed by oxygen flask combustion. The linear range was 10-500 ng mL-1 for plasma and 10-100 ng mL-1 for hair. Within-day and day-to-day precisions for plasma ranged from 5.4% to 9.3% (n = 7) and from 3.5% to 14.5% (n = 7), respectively. Within-day precisions for hair ranged from 0.6% to 6.2% (n = 7). Recoveries ranged from 91.0% to 97.8% for plasma and from 95.0% to 102.0% for hair. The blood samples from 15 Hans and 20 Uygurs in Xinjiang Uygur automatic region and 23 Hans in Liaoning province were collected and determined. It was indicated that no statistically significant difference in plasma selenium concentration of the Hans between Xinjiang and Liaoning was found (F = 1.36, P > 0.05). However, there were statistically significant differences between the Hans and the Uygurs in Xinjiang (F = 1.01, P < 0.01) and between males and females in the two areas (P < 0.01). There was a low correlation between plasma selenium concentration and hair selenium concentration. The ratio of hair selenium concentration to plasma selenium concentration was 2.17, with a range of 1.63-2.88.

Hair↗
PubMed1976

Studies by electron-paramagnetic-resonance spectroscopy and stopped-flow spectrophotometry on the mechanism of action of turkey liver xanthine dehydrogenase.

Studies by e.p.r. (electron-paramagnetic-resonance) spectroscopy and by stopped-flow spectrophotometry on turkey liver xanthine dehydrogenase revealed strong similarities to as well as important differences from the Veillonella alcalescens xanthine dehydrogenase and milk xanthine oxidase. The turkey enzyme is contaminated by up to three non-functional forms, giving molybdenum e.p.r. signals designated Resting I, Resting II and Slow. Slow and to a lesser extent Resting I signals are like those from the Veillonella enzyme, whereas Resting II is very like a resting signal described by K. V. Rajagopolan, P. Handler, G. Palmer & H. Beinert (1968) (J. Biol. Chem. 243, 3784-3796) for aldehyde oxidase. Another non-functional form that gives the Inhibited signal is produced on treatment of the enzyme with formaldehyde. Stopped-flow measurements at 450 nm show that, as for the milk enzyme, reduction by xanthine is rate-limiting in enzyme turnover. The active enzyme gives rise to Very Rapid and Rapid molybdenum(V) e.p.r. signals, as well as to an FADH signal. That these signals are almost indistinguishable from those of the milk enzyme, confirms the similarities between the active sites. There are two types of iron-sulphur centres that give signals like those in the milk enzyme, though with slightly different parameters. Quantitative reduction titration of the functional enzyme with xanthine revealed two important differences between the turkey and the milk enzymes. First, the turkey enzyme FADH/FADH2 system has a redox potential sufficiently low that xanthine is incapable of reducing the flavin completely. This finding presumably explains the very low oxidase activity. Secondly, whereas the Fe/S II chromophore in the milk enzyme has a relatively high redox potential, for the turkey enzyme the value of this potential is lower and similar to that of its Fe/S I chromophore.

Animals↗
PubMed2000

Optimizing cerebral oxygenation during cardiac surgery: a review of experimental and clinical investigations with near infrared spectrophotometry.

OBJECTIVES: Near-infrared spectrophotometry (NIRS) is a promising method for non-invasive monitoring of cerebral oxygenation and hemodynamics. This paper reviews studies in which we aimed to validate NIRS in an experimental model of cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) (validation study), use the method in experimental settings to optimize cerebral oxygenation during CPB (oxygenation study), and test its utility during routine cardiac surgery (clinical study). METHODS: Validation study: Forty 8-10 kg piglets underwent 60 min of DHCA at 15 degrees C, were extubated on the first postoperative day, and sacrificed on the 4th postoperative day for histologic investigations. During CPB and DHCA, the animals were investigated by NIRS (monitoring of cerebral oxygenated hemoglobin (HbO2) and oxidized cytochrome aa3 (Cytox)) and magnetic resonance spectroscopy (MRS) (monitoring of cerebral adenosine triphosphate (ATP) and phosphocreatine (PCr). Oxygenation study: A normoxic (n = 5) and a hyperoxic group (n = 5) of piglets underwent 120 min of DHCA and 6 h of reperfusion with NIRS monitoring. Neuronal damage was evaluated by histology. Clinical study: Patients (n = 41) undergoing routine cardiac surgery were investigated by NIRS and neuropsychological testings. RESULTS: Validation study: Reductions of CytOx and HbO2 values were closely correlated with decreases in ATP, PCr, and pHi. The changes in CytOx and PCr showed the strongest correlation (r = 0.623). Maximal CytOx reduction during DHCA predicted brain damage. Oxygenation study: Histology revealed a significant increase in brain damage in the normoxic group. Cytox and HbO2 tended to be lower during DHCA (p = 0.16). Clinical study: During CPB, arterial PCO2, pH and temperature were closely related to CytOx and HbO2. Patients who suffered from neuropsychological deficits had a lower CytOx minimum compared to those without these. CONCLUSIONS: NIRS measurements, especially the CytOx signal, correlate well with high energy phosphates and have a high sensitivity to predict histologic brain damage. NIRS is an important and useful tool in studies investigating cerebral oxygenation during CPB. The CytOx signal predicted impaired neuropsychological outcome in patients. Therefore, the CytOx signal holds much interest for future studies.

Animals↗
PubMed2002

A method for determining crystalline silica in bulk samples by Fourier transform infrared spectrophotometry.

A method for determining crystalline silica (alpha-quartz) in bulk samples has been developed. This procedure requires the bulk sample to be homogenized and sieved down to particle sizes <10 micro m in diameter. The obtained fraction is analysed by Fourier transform infrared spectrophotometry. Method validation was carried out using 55 real-world bulk samples, containing between 1 and 75% (w/w) alpha-quartz. The method is capable of quantifying alpha-quartz down to 1% (w/w) in routine analysis. However, the method is unlikely to allow routine quantification at 0.1% (w/w) as currently mandated by regulations.

Particle Size↗
PubMed2002

Ictal cerebral haemodynamics of childhood epilepsy measured with near-infrared spectrophotometry.

Near-infrared spectrophotometry (NIRS) is a new technique that allows continuous non-invasive monitoring of tissue oxygenation and haemodynamics in the brain. We used NIRS in various types of paediatric epileptic seizures in order to understand the pathophysiology of epileptic seizures in childhood epilepsy. This study examined 15 children ranging in age from 1.5 months to 16 years (nine males and six females), with different types of epilepsy. Six series of tonic spasms and 67 isolated seizures were recorded. The results demonstrated that several pathophysiological processes exist during epileptic seizures in childhood. (i) Convulsive seizures were associated with a remarkable increase in cerebral blood volume (CBV), while absence seizures were associated with a mild decrease or no change in CBV of the frontal cortex. (ii) An initial transient decrease in CBV was observed in some types of convulsive seizures. (iii) An ictal increase in CBV changed to an ictal decrease in the course of tonic status epilepticus. (iv) There was definite heterogeneity in the CBV changes during tonic spasms in patients with West syndrome. NIRS is easily applicable to paediatric patients with epilepsy and may provide new insights into the pathophysiology of various types of epileptic seizure.

Adolescent↗
PubMed1991

Benzo[a]pyrene-globin adducts detected by synchronous fluorescence spectrophotometry: method development and relation to lung DNA adducts in mice.

A simple synchronous fluorescence spectrophotometry (SFS) to detect benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-globin adducts is described. SFS for BPDE-DNA, which measures detached benzo[a]pyrene (B[a]P)-tetrols after acid hydrolysis of DNA, was applied for BPDE-globin adducts in B[a]P-treated C57BL/6 (B6) mice. Unlike DNA samples, globin is not measurable as such after acid hydrolysis because proteins give a background in SFS. Furthermore, proteinase incubation before acid hydrolysis of globin gave too much background even after purification to be useful in this assay. Of several purification procedures tried after acid hydrolysis (protein precipitation, elution through Sep-Pak C18, filtration, ether extraction of tetrols), the lowest background fluorescence was obtained with ether extractions of B[a] moieties. Ether phases were evaporated to dryness and the remainder dissolved in distilled water (1 ml), which was measured by SFS. Compared to DNA, somewhat milder hydrolysis conditions were optimal for globin samples (0.05 M HCl, 1.5 h, + 90 degrees C). Globin samples from B[a]P-treated mice gave a peak at the same wavelength (345 nm excitation) as the hydrolysis products of BPDE-DNA adducts, indicating B[a]P-tetrols and triols in the sample. Less than half of B[a]P measured in globin was from covalently bound BPDE. In mice injected i.p. with 1-160 mg/kg of B[a]P there was a dose-dependent increase in the amount of BPDE adducts in globin and a positive correlation with lung and liver DNA. Globin adducts were a more sensitive indicator of B[a]P exposure than DNA adducts because more globin can be used for the assay. Although both covalently and non-covalently bound BPDE in globin are detected by SFS, this method is the simplest described so far, reproducible and theoretically sensitive enough for human biomonitoring.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMedDate not supplied

Near infrared spectrophotometry reflects cerebral metabolism during hypothermic circulatory arrest in adults.

Near-infrared spectrophotometry (NIRS) is assumed to reflect cerebral oxygenation during hypothermic circulatory arrest (HCA). However, the rationale for the use of NIRS as a marker of cerebral metabolism remains to be elucidated. We examined whether cerebral oxygenation measured by NIRS correlated with cerebral metabolic function assessed by cerebral oxygen extraction ratio (OER) during HCA in adults. NIRS was continuously monitored in 14 patients undergoing thoracic aortic surgery using HCA (17.9 +/- 2.9 degrees C esophageal temperature at HCA onset). Jugular venous oxygen saturation and OER were also monitored. OER was significantly reduced from 37.3 +/- 7.2% at the onset of cardiopulmonary bypass to 13.6 +/- 6.2% at the onset of HCA (p < 0.0001). A linear decrease in oxygenated-hemoglobin (oxy-Hb) and increase in deoxygenated-hemoglobin (deoxy-Hb) were found during HCA, which returned to baseline levels after rewarming. The rate of decrease in oxy-Hb and increase in deoxy-Hb were -0.63 +/- 0.45 and 0.51 +/- 0.30 (10(-5) OD/cm per sec), respectively, both of which significantly correlated with OER at the onset of HCA (R2=0.739 and 0.633; p < 0.0001 and p = 0.0007, respectively). NIRS may serve as a reliable diagnostic modality for monitoring cerebral metabolism during aortic surgery using HCA.

Adult↗
PubMed1971

Red and far red effects on phenylalanine ammonia-lyase in raphanus and sinapis seedlings do not correlate with phytochrome spectrophotometry.

In seedlings of Raphanus sativus (radish) and Sinapis alba (mustard), irradiation for 6 hours with far red light significantly increases the extractable activity of phenylalanine ammonia-lyase by the end of the light period. A schedule of 10 minutes red light-110 minutes darkness-10 minutes red-110 minutes darkness-10 minutes red-110 minutes darkness has no effect as compared to dark controls. However, the red light program maintains a level of far red-absorbing phytochrome always measurable by in vivo spectrophotometry during the 6-hour experimental period. We conclude that the far red effect on this enzyme and for this specific material cannot be explained solely by formation and maintenance of far red-absorbing phytochrome.

Journal Article↗
PubMed1991

Measurement of legume nodule respiration and o(2) permeability by noninvasive spectrophotometry of leghemoglobin.

Physiological regulation of nodule gas permeability has a central role in the response of legumes to such diverse factors as drought, defoliation, and soil nitrate. A new method for quantifying nodule respiration and O(2) permeability, based on noninvasive spectrophotometry of leghemoglobin, was evaluated using intact, attached nodules of Lotus corniculatus. First, the relationship between nodule respiration (O(2) consumption) rate and internal O(2) concentration was determined from the rate of decrease in fractional oxygenation of leghemoglobin (FOL) under N(2). The rate of increase of FOL under 100% O(2) was then used to calculate nodule O(2) permeability, after correcting for respiration. Inactivation of nitrogenase by exposure to 100% O(2) for 15 minutes led to decreases in both permeability and O(2)-saturated respiration (V(max)), but the brief (<15 seconds) exposures to 100% O(2) required by the assay itself had little effect on either parameter. A gradual increase in external O(2) concentration from 20 to 40% resulted in a reversible decrease in permeability, but no change in V(max). The new method is likely to be useful for research on nodule physiology and might also be applicable to agronomic research and crop improvement programs.

Journal Article↗
PubMed2002

Comparison of three methods to measure absolute cerebral hemoglobin concentration in neonates by near-infrared spectrophotometry.

Three methods by which to determine absolute total cerebral hemoglobin concentration (tHb in micromol/L) by near-infrared spectrophotometry (NIRS) have evolved: (1) tHbo, requiring oxygenation changes and arterial oxygen saturation measurements as a reference using a relative NIRS algorithm, (2) tHbg, using a geometrical multidistance principle and (3) tHbgo, a combination of both. The aim of this study was to compare the three methods quantitatively. Sixteen clinically stable preterm infants with a mean gestational age of 29.6 (range of 25.1-36.4) weeks, birthweight of 1386 (680-2820) g and a postnatal age of 2.5 (0.5-6) days, who needed supplemental oxygen, were enrolled. The mean+/-standard deviation tHbg was 150.2+/-41.8 micromol/L (range of 61.6-228.9 micromol/L), the tHbo was 62.1+/-27.2 micromol/L (26.0-110.8 micromol/L) and the tHbgo was 89.3+/-45.6 micromol/L (26.5-195.9 micromol/L). The correlation coefficient among the three methods were tHbg and tHbgo r=0.736; tHbo and tHbgo r=0.938; tHbg and tHbo r=0.598. A multiple regression with variable selection by Mellow's C(p) showed, that tHbg was correlated to the birthweight, the postnatal age, the heart rate and the pCO2 (r(2)=0.588), tHbo and tHbgo were associated with the hemoglobin concentration in the blood, the mean arterial blood pressure and the pCO2 (r(2)=0.493 and 0.406, respectively). The three methods (tHbg, tHbo, and tHbgo) give systematically different tHb readings and large intersubject variability.

Brain Chemistry↗
PubMed1999

CO2 reactivity of the cerebral hemoglobin concentration in healthy term newborns measured by near infrared spectrophotometry.

CO2 reactivity of cerebral hemoglobin concentration was studied in 16 healthy term neonates on days 1 and 4 after birth using the near infrared spectrophotometry (NIRS) technique. The aim was to establish data on the physiological range of CO2 reactivity in healthy newborns and to investigate the influence of postnatal age on it. The CO2 reactivity measured by NIRS is expressed as the change of the total cerebral hemoglobin concentration (tHbR) per change of CO2 tension in micromol/l/kPa. We evaluated CO2 reactivity during increases and decreases of transcutaneous CO2 partial pressure and found in our methodological setting the data of the increases more reliable. In all infants but 1 we found a tHbR on day 1 with a mean value of 8.19 micromol/l/kPa (-1.39 to 18.87), in all infants on day 4 with a mean value of 9.54 micromol/l/kPa (2.76-25. 88). There is a trend to higher values between day 1 and day 4 (difference = 2.25 micromol/l/kPa; p = 0.08). The noninvasive NIRS technique enabled us to test the cerebrovascular CO2 reactivity of the tHbR for the first time in healthy term newborns. Data on its physiologic range and variability are presented and compared to findings from ventilated infants and other age groups. As the CO2 reactivity might be an indicator for infants at risk of cerebral damage, it is necessary to have data on the physiological range of this parameter.

Brain↗