Let's talk about PFAS: Inconsistent public awareness about PFAS and its sources in the United States.

The presence of per- and polyfluoroalkyl substances (PFAS) in U.S. drinking water has recently garnered significant attention from the media, federal government, and public health professionals. While concerns for PFAS exposure continue to mount, the general public's awareness and knowledge of the contaminant has remained unknown. This exploratory study sought to fill this data gap by administering a nationwide survey in which the awareness of PFAS and community contamination, awareness of PFAS containing products and intentions to change product use, and awareness and concern about PFAS in drinking water were assessed. The results indicated that almost half the respondents had never heard of PFAS and do not know what it is (45.1%). Additionally, 31.6% responded that they had heard of PFAS but do not know what it is. A large portion of respondents (97.4%) also responded that they did not believe their drinking water had been impacted by PFAS. Demographic association did not influence knowledge of PFAS or levels of concern with PFAS in drinking water. The strongest predictor of PFAS awareness was awareness due to known community exposure. The respondents aware of community exposure were more likely to have knowledge of PFAS sources, change their use of items with potential PFAS contamination, and answer that their drinking water sources were also contaminated with PFAS. Based on the received responses, PFAS information and health risks need to be better communicated to the public to help increase awareness. These efforts should also be coordinated between government agencies, utilities, the research community, and other responsible entities to bolster their effectiveness.

HNA antibody-mediated neutrophil aggregation is dependent on serine protease activity.

BACKGROUND AND OBJECTIVES: Transfusion-related acute lung injury (TRALI) is often caused by antibodies against human neutrophil alloantigen-2 (HNA-2) and HNA-3a. Neutrophil aggregation is considered as a major cause of TRALI, but little is known about how HNA antibodies initiate this process. We explored mechanisms involved in neutrophil aggregation induced by HNA-2 and HNA-3a antibodies. MATERIALS AND METHODS: Isolated neutrophils were pretreated with broad-spectrum or specific inhibitors against different cell functions or proteases. Granulocyte agglutination test (GAT) was performed with serially diluted anti-HNA-2 and anti-HNA-3a plasmas or control plasma, and reactivity was evaluated microscopically. Reactive oxygen species (ROS) production in neutrophils was investigated using a lucigenin-based chemiluminescence assay. RESULTS: HNA-2 and HNA-3a antibody-mediated neutrophil aggregation was inhibited by pretreatment with formaldehyde, iodoacetamide and the serine protease inhibitors Pefabloc-SC, N-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and Nα-tosyl-L-lysine chloromethyl ketone hydrochloride (TLCK). In contrast, inhibition of actin polymerization, respiratory burst, cysteine proteases, metalloproteases or aspartic proteases did not affect neutrophil aggregation. Furthermore, HNA-3a antibodies did not directly cause ROS production in neutrophils. CONCLUSION: Aggregation of neutrophils induced by HNA-2 and HNA-3a antibodies is an active process and depends on trypsin- or chymotrypsin-like serine proteases but is not dependent on the production of ROS. These findings may open new prospects for the pharmacologic prevention of neutrophil-associated acute lung injury.

Probing the photoexcited states of rhodium corroles by time-resolved Q-band EPR. Observation of strong spin-orbit coupling effects.

The photoexcited states of two 5,10,15-tris(pentafluorophenyl)corroles (tpfc), hosting Rh(III) in their core, namely Rh(pyr)(PPh 3)(tpfc) and Rh(PPh 3)(tpfc), have been studied by time-resolved electron paramagnetic resonance (TREPR) combined with pulsed laser excitation. Using the transient nutation technique, the spin polarized spectra are assigned to photoexcited triplet states. The spectral widths observed for the two Rh(III) corroles crucially depend on the axial ligands at the Rh(III) metal ion. In case of Rh(PPh 3)(tpfc), the TREPR spectra are found to extend over 200 mT, which exceeds the spectral width of non-transition-metal corroles by more than a factor of 3. Moreover, the EPR lines of the Rh(III) corroles are less symmetric than those of the non-transition-metal corrroles. The peculiarities in the TREPR spectra of the Rh(III) corroles can be rationalized in terms of strong spin-orbit coupling (SOC) associated with the transition-metal character of the Rh(III) ion. It is assumed that SOC in the photoexcited Rh(III) corroles effectively admixes metal centered (3)dd-states to the corrole centered (3)pipi*-states detected in the TREPR experiments. This admixture leads to an increased zero-field splitting and a large g-tensor anisotropy as manifested by the excited Rh(III) corroles.

Exposure to pulse-modulated radio frequency electromagnetic fields affects regional cerebral blood flow.

We investigated the effects of radio frequency electromagnetic fields (RF EMF) similar to those emitted by mobile phones on waking regional cerebral blood flow (rCBF) in 12 healthy young men. Two types of RF EMF exposure were applied: a 'base-station-like' and a 'handset-like' signal. Positron emission tomography scans were taken after 30 min unilateral head exposure to pulse-modulated 900 MHz RF EMF (10 g tissue-averaged spatial peak-specific absorption rate of 1 W/kg for both conditions) and sham control. We observed an increase in relative rCBF in the dorsolateral prefrontal cortex on the side of exposure. The effect depended on the spectral power in the amplitude modulation of the RF carrier such that only 'handset-like' RF EMF exposure with its stronger low-frequency components but not the 'base-station-like' RF EMF exposure affected rCBF. This finding supports our previous observation that pulse modulation of RF EMF is necessary to induce changes in the waking and sleep EEG, and substantiates the notion that pulse modulation is crucial for RF EMF-induced alterations in brain physiology.

Electromagnetic fields, such as those from mobile phones, alter regional cerebral blood flow and sleep and waking EEG.

Usage of mobile phones is rapidly increasing, but there is limited data on the possible effects of electromagnetic field (EMF) exposure on brain physiology. We investigated the effect of EMF vs. sham control exposure on waking regional cerebral blood flow (rCBF) and on waking and sleep electroencephalogram (EEG) in humans. In Experiment 1, positron emission tomography (PET) scans were taken after unilateral head exposure to 30-min pulse-modulated 900 MHz electromagnetic field (pm-EMF). In Experiment 2, night-time sleep was polysomnographically recorded after EMF exposure. Pulse-modulated EMF exposure increased relative rCBF in the dorsolateral prefrontal cortex ipsilateral to exposure. Also, pm-EMF exposure enhanced EEG power in the alpha frequency range prior to sleep onset and in the spindle frequency range during stage 2 sleep. Exposure to EMF without pulse modulation did not enhance power in the waking or sleep EEG. We previously observed EMF effects on the sleep EEG (A. A. Borbély, R. Huber, T. Graf, B. Fuchs, E. Gallmann and P. Achermann. Neurosci. Lett., 1999, 275: 207-210; R. Huber, T. Graf, K. A. Cote, L. Wittmann, E. Gallmann, D. Matter, J. Schuderer, N. Kuster, A. A. Borbély, and P. Achermann. Neuroreport, 2000, 11: 3321-3325), but the basis for these effects was unknown. The present results show for the first time that (1) pm-EMF alters waking rCBF and (2) pulse modulation of EMF is necessary to induce waking and sleep EEG changes. Pulse-modulated EMF exposure may provide a new, non-invasive method for modifying brain function for experimental, diagnostic and therapeutic purposes.

Structure of the P700(+ )A1(-) radical pair intermediate in photosystem I by high time resolution multifrequency electron paramagnetic resonance: analysis of quantum beat oscillations.

The geometry of the secondary radical pair P700(+)A1(-), in photosystem I (PSI) from the deuterated and 15N-substituted cyanobacterium Synechococcus lividus, has been determined by high time resolution electron paramagnetic resonance (EPR), performed at three different microwave frequencies. Structural information is extracted from light-induced quantum beats observed in the transverse magnetization of P700(+)A1(-) at early times after laser excitation. A computer analysis of the two-dimensional Q-band experiment provides the orientation of the various magnetic tensors of with respect to a magnetic reference frame. The orientation of the cofactors of the primary donor in the g-tensor system of is then evaluated by analyzing time-dependent X-band EPR spectra, extracted from a two-dimensional data set. Finally, the cofactor arrangement of P700(+)A1(-) in the photosynthetic membrane is deduced from angular-dependent W-band spectra, observed for a magnetically aligned sample. Thus, the orientation of the g-tensor of P700(+) with respect to a chlorophyll based reference system could be determined. The angle between the g1(z) axis and the chlorophyll plane normal is found to be 29 +/- 7 degrees, while the g1(y) axis lies in the chlorophyll plane. In addition, a complete structural model for the reduced quinone acceptor, A1(-), is evaluated. In this model, the quinone plane of is found to be inclined by 68 +/- 7 degrees relative to the membrane plane, while the P700(+)-A1(-) axis makes an angle of 35 +/- 6 degrees with the membrane normal. All of these values refer to the charge separated state, observed at low temperatures, where forward electron transfer to the iron-sulfur centers is partially blocked. Preliminary room temperature studies of P700(+)A1(-), employing X-band quantum beat oscillations, indicate a different orientation of A1(-) in its binding pocket. A comparison with crystallographic data provides information on the electron-transfer pathway in PSI. It appears that quantum beats represent excellent structural probes for the short-lived intermediates in the primary energy conversion steps of photosynthesis.

Tetrahydrobiopterin increases myocardial blood flow in healthy volunteers: a double-blind, placebo-controlled study.

OBJECTIVES: Tetrahydrobiopterin (BH4) is a regulatory cofactor for the activity of nitric oxide synthases. Vasodilating properties of BH4 have been reported in vitro and in vivo. The influence of BH4 on myocardial blood flow (MBF), however, is largely unknown. We therefore performed a double-blind, placebo-controlled study to investigate the effect of intravenous BH4 on MBF in healthy volunteers. METHODS AND RESULTS: Resting MBF was assessed in 15 subjects receiving either intravenous BH4 (10 mg/kg) or placebo using positron emission tomography (PET) and [13N]ammonia. From a mean baseline MBF of 0.91 +/- 0.09 ml/min/g, MBF increased to 1.18 +/- 0.10 ml/min/g after BH4 (n = 10; p = 0.0042). In contrast, in the group receiving placebo mean MBF remained unchanged (non-significant decrease from 0.97 +/- 0.19 to 0.84 +/- 0.11 ml/min/g; n = 5; p = 0.36). Systemic haemodynamics and ECGs remained unaffected in both groups. BH4 was very well tolerated. CONCLUSION: Systemically administered BH4 is safe and effectively increases resting MBF in healthy volunteers.

Increased myocardial blood flow during acute exposure to simulated altitudes.

BACKGROUND: Although only poor data exist on changes in myocardial blood flow (MBF) under acute hypoxia, patients with known coronary artery disease are advised not to exceed a moderate altitude exposure of about 2000 m above sea level. METHODS AND RESULTS: We measured MBF with positron emission tomography using O-15--labeled water in 8 healthy human volunteers (aged 26 +/- 3 years [mean +/- SD]) at baseline (450 m above sea level, Zurich, Switzerland) and during acute hypoxic hypoxemia induced by inhalation of 2 hypoxic gas mixtures corresponding to altitudes of 2000 and 4500 m. MBF remained unchanged at 2000 m (increase of 10%, not significant) but increased significantly at 4500 m (62%, P <.001), exceeding the relative increase in rate pressure product. CONCLUSIONS: Our results may explain why exposure to an altitude of 2000 m (corresponding to the cabin pressure in most airplanes during flight) is clinically well tolerated, even by patients with reduced coronary flow reserve, such as those with coronary artery disease. However, at an altitude of 4500 m, MBF increases significantly, supporting the recommendation that patients with impaired flow reserve avoid exposure to higher altitudes.

No difference in brain activation during cognitive performance between ecstasy (3,4-methylenedioxymethamphetamine) users and control subjects: a [H2(15)O]-positron emission tomography study.

The long-term use of the serotonin-releaser and uptake-inhibitor 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") has been associated with memory impairments and increased liability to depressive mood and anxiety attacks. It is unclear, however, whether these psychologic deviations are reflected in alterations of the underlying neurophysiologic substrate. The authors compared mood and regional cerebral blood flow (rCBF) profiles between regular polytoxic Ecstasy users and Ecstasy-naive controls. Brain activity as indexed by rCBF was measured during cognitive activation by an attentional task using positron emission tomography and [H2(15)O]. Mood was assessed by means of the Hamilton Rating Scale for Depression (HAM-D) and the EWL Mood Rating Scale. Statistical parametric mapping revealed that brain activity did not differ between the two groups. Both groups also performed equally on the cognitive task requiring sustained attention. However, significantly higher levels of depressiveness as determined by the HAM-D and EWL scales were found in Ecstasy-using subjects. These data indicate that, despite differences in mood, polytoxic Ecstasy users do not differ from Ecstasy-naive controls in terms of local brain activity. Heightened depressiveness in the Ecstasy group was consistent with results from previous studies and could be related to serotonergic hypofunction resulting from repeated MDMA consumption. However, this study cannot exclude the possibility that the observed differences are preexisting rather than a result of Ecstasy use.

3,4-Methylenedioxymethamphetamine (MDMA) modulates cortical and limbic brain activity as measured by [H(2)(15)O]-PET in healthy humans.

[H(2)(15)O]-Positron Emission Tomography (PET) was used to examine regional cerebral blood flow (rCBF) after administration of a single oral dose of the serotonin realeaser and uptake inhibitor MDMA (1.7 mg/kg) or placebo to 16 MDMA-naïve subjects. Psychological changes were assessed by psychometric rating scales. MDMA produced distributed changes in regional blood flow including increases in ventromedial frontal and occipital cortex, inferior temporal lobe and cerebellum; and decreases in the motor and somatosensory cortex, temporal lobe including left amygdala, cingulate cortex, insula and thalamus. Concomitant with these changes, subjects experienced heightened mood, increased extroversion, slight derealization and mild perceptual alterations. MDMA also produced increases in blood pressure and several side effects such as jaw clenching, lack of appetite and difficulty concentrating. These results indicate that a distributed cluster of brain areas underlie the various effects of MDMA in humans.

Radiation exposure to sonographers from fluorine-18-FDG PET patients.

OBJECTIVE: We estimated the amount of radiation exposure to sonographers from patients who were injected with 18F-fluorodeoxyglucose (FDG) at 2 and 3 h postinjection. METHODS: We studied 8 patients who were given between 380-420 MBq 18F-FDG. The patients were measured with a RADOS RDS-120 dosimeter between 2 and 3 h after FDG injection. The dosimetry measurement was taken at a distance of 0.5 m from the injected patient, a distance used by a sonographer to perform an abdominal ultrasound. Measurements were taken at the levels of the sonographer's shoulder, abdomen, and gonads. RESULTS: At the first measurement at 2 h, the mean exposures to the shoulder, abdomen, and gonads of the sonographer in pSv/h were 31.9+/-11.3, 37.1+/-9.5, and 32.8+/-11.8, respectively. At 3 h, the mean exposures to the shoulder, abdomen, and gonads were 21.5+/-4.2, 20.2+/-5.8, and 19.6+/-4.9, respectively. CONCLUSION: The amount of radiation exposure to a sonographer is minimal. Radiation exposure risks should be considered, however, if the sonographer comes into daily, repeated contact with patients who have been given 18F-FDG.

Functional neuroanatomy of human sleep states after zolpidem and placebo: a H215O-PET study.

Changes in the functional organization of the brain during the course of sleep and waking are reflected by different patterns of regional cerebral blood flow (rCBF). To investigate the effect of the hypnotic zolpidem, a benzodiazepine receptor agonist, drug or placebo were administered to eight young, healthy men prior to bedtime. The subjects were sleep-deprived to promote sleep during the 4-h recording period in the positron emission tomography scanner. Intravenous injections of labelled water were administered during pre-drug wakefulness, and during Stage 2, Stage 4 and rapid eye movement (REM) sleep, each injection being followed by an emission scan. Statistical parametric mapping was used to investigate the effects of treatment and sleep states. During sleep (combined Stages 2 and 4, and REM sleep) relative rCBF was lower after zolpidem than after placebo in the basal ganglia and insula, and higher in the parietal cortex. A 'multiple study' analysis of REM sleep revealed that rCBF in the anterior cingulum was lower after zolpidem than after placebo, whereas rCBF in the occipital and parietal cortex, parahippocampal gyrus and cerebellum was higher. When the pooled data (drug and placebo) of Stages 2 and 4 were compared with wakefulness, rCBF was lower in prefrontal cortex and insula, and higher in the occipital and parietal cortex. The results indicate that some differences in rCBF from wakefulness to non-REM sleep are further augmented by zolpidem.

Non-small cell lung cancer: nodal staging with FDG PET versus CT with correlative lymph node mapping and sampling.

PURPOSE: To prospectively compare the accuracy of positron emission tomography (PET) with 2-[fluorine-18] fluoro-2-deoxy-D-glucose (FDG) with that of computed tomography (CT) in the nodal staging of non-small cell lung cancer. MATERIALS AND METHODS: PET and contrast material-enhanced CT were performed in 47 patients suspected of having or with newly diagnosed non-small cell lung cancer. Each imaging study was evaluated separately, and nodal stations were localized according to the American Thoracic Society mapping system. Extensive lymph node sampling (599 nodes from 191 nodal stations) of the ipsi- and contralateral tracheobronchial and mediastinal nodal stations was performed at thoracotomy and/or mediastinoscopy. Imaging findings were correlated with histopathologic staging results. RESULTS: The sensitivity of PET and CT was 89% and 57%, respectively, for the staging of N2 or N3 disease in mediastinal nodes; specificity was 99% and 94%, respectively; positive predictive value was 96% and 76%, respectively; negative predictive value was 97% and 87%, respectively; and accuracy was 96% and 85%, respectively. In assigning the correct N stage, PET was correct in 96% and CT in 79% of cases. CONCLUSION: PET with FDG appears to be superior to CT for nodal staging of non-small cell lung cancer.

Whole-body PET: physiological and artifactual fluorodeoxyglucose accumulations.

UNLABELLED: The purpose of this study was to semiquantitatively identify artifactual and physiological soft-tissue accumulations in whole-body FDG-PET scans with the aim of defining their frequency and anatomic distribution. METHODS: Fifty whole-body FDG-PET scans performed for the staging of malignant melanoma were obtained from transaxial scans and reconstructed without absorption correction by filtered backprojection in the form of coronal and sagittal sections. The patients were asked to stay n.p.o. for at least 4 hr and interrogated about their physical activity prior to injection and until scanning. Classification of FDG organ accumulations was done using grades 0-6. Means and standard deviations on this scale were then calculated for multiple organs and muscle groups and tabulated. RESULTS: On this grading scale, viscera showed uptake grades between 1.7 +/- 0.5 and 2.05 +/- 1.0. Except for the intestines, the activity in these organs was homogeneously distributed. Relatively high average uptake values of 2.0-4.2 (s.d. > or = 2.3) were found in various muscle groups, especially the orbital musculature. Myocardial uptake was visible in 90% of the scans. Reconstruction artifacts were seen around the renal collecting system and the bladder. CONCLUSION: Most of the "normal" accumulations of FDG in nonattenuation corrected whole-body PET are readily recognized and distinct from the usually focal FDG accumulation associated with metastatic disease, but the diagnostician must be familiar with them. Muscular FDG uptake is related to physical activity prior and immediately following injection and can be minimized by proper patient instructions and positioning.

Malignant melanoma: staging with whole-body positron emission tomography and 2-[F-18]-fluoro-2-deoxy-D-glucose.

PURPOSE: To evaluate whole-body positron emission tomography (PET) with 2-[fluorine-18]-fluoro-2-deoxy-D-glucose (FDG) in the detection of metastasis from melanoma. MATERIALS AND METHODS: Whole-body PET was performed in 33 patients with either known metastatic or newly diagnosed melanoma. Patients with suspected metastases also underwent computed tomography, magnetic resonance imaging, or both. Diagnoses were confirmed with histologic examination or with at least one imaging modality in addition to PET. Blinded interpretations of PET scans were performed. RESULTS: Forty of 53 lesions evaluated proved to be melanoma metastases. Whole-body PET correctly depicted 37 sites of metastases. Three cutaneous metastases (< 3 mm) were missed. PET correctly excluded malignancy in 10 cases where suspicious lesions were found with conventional cross-sectional imaging modalities but later ruled out with fine-needle biopsy. In six patients, PET depicted new metastases. The sensitivity for the detection of malignant lesions was 92%; the specificity for reading the PET images without clinical information was 77% and with clinical information was 100%. CONCLUSIONS: These results suggest that whole-body FDG PET is an effective imaging modality to screen for metastases from malignant melanoma.

Staging of metastatic melanoma by whole-body positron emission tomography using 2-fluorine-18-fluoro-2-deoxy-D-glucose.

Metastatic melanoma was staged in 15 patients using whole-body positron emission tomography (PET) and the radiopharmaceutical 2-fluorine-18-fluoro-2-deoxy-D-glucose (FDG). PET correctly demonstrated 30 metastases in lung, brain, pancreas, nasal cavity, skin and subcutaneous tissue, and lymph nodes. It detected 97% of all metastases exceeding its spatial resolution (> 5 mm). Two cutaneous metastases (approximately 3 mm) did not show increased FDG uptake; the overall detection sensitivity was 91%. Two false-positive lesions in one patient were due to severe wound infection. PET correctly excluded malignancy in four cases where suspicious lesions were found with conventional cross-sectional imaging modalities but later ruled out by fine-needle biopsy. PET therefore proved to be an excellent method for staging of metastatic melanoma. Due to its high sensitivity for malignant lesions and the possibility of covering the whole body in one examination, it can replace staging techniques employing multiple imaging modalities: chest X-ray, ultrasonography and computed tomography. Furthermore, it provides information on the malignant potential of the detected lesion.

Dual-isotope SPET 201Tl rest/99mTc-MIBI stress: one-day protocol for pre-operative myocardial imaging?

We compared the results of a rest 201TI/stress 99mTc-MIBI protocol, both by means of separate single isotope and simultaneous dual isotope acquisition, with a standard stress/rest 99mTc-MIBI 2-day protocol in 11 patients with low probability of CAD and 14 patients with chronic CAD. In patients with CAD 406 segments (sgs) were analysed. In the standard protocol 119 sgs were classified as pathological of which 50.4% were fixed and 49.6% reversible defects. With the MIBI-stress/TI-rest single 33% of 119 pathological sgs were fixed and 67% reversible defects. With the MIBI-stress/TI-rest dual only 20% were fixed and 80% reversible defects. The reversibility of 59 MIBI-stress/MIBI-rest reversible defects was quantified: MIBI-stress/MIBI-rest 35 +/- 16% MIBI-stress/TI-rest single 50 +/- 26% and MIBI-stress/TI-rest dual 48 +/- 22%. The results of 99mTc-MIBI rest and 201TI rest studies in patients with chronic CAD are not the same. Dual-isotope 1-day 201TI-rest/99mTc-MIBI-stress SPET data, acquired separately, may give fast and complete information on myocardial perfusion at stress and rest, respectively, and on myocardial viability.

[Hydrophobic acridine dyes for fluorescent staining of mitochondria in living cells. 3. Specific accumulation of the fluorescent dye NAO on the mitochondrial membranes in HeLa cells by hydrophobic interaction. Depression of respiratory activity, changes in the ultrastructure of mitochondria due to NAO. Increase of fluorescence in vital stained mitochondria in situ by irradiation]. / Uber hydrophobe Acridinfarbstoffe zur Fluorochromierung von Mitochondrien in lebenden Zellen. 3. Mitteilung: Spezifische Akkumulation des Farbstoffs NAO an die Mitochondrienmembranen von HeLa-Zellen durch hydrophobe Wechselwirkung. Hemmung der Atmungsaktivität, Veränderung der Ultrastruktur der Mitochondrien durch NAO. Intensivierung der Fluoreszenz vital gefärbter Mitochondrien in situ bei Bestrahlen.

The hydrophobic fluorescence dye 10-n-nonyl-acridinium-orange-chloride, NAO, stains specifically the mitochondria of living HeLa-cells. A dye concentration of 1 X 10(-8) M is sufficient for vital staining and at 5 X 10(-7) M an incubation time less than 1 min is enough to generate the bright green fluorescence of the mitochondria. The retention of NAO by the mitochondria is longer than 7 days. The dye accumulation is not affected by the ionophores valinomycin, nigericin, gramicidin, the uncoupling agents DNP, CCCP or by ouabain. In contrast to Rh 123 the trans-membrane potential is not the driving force of the NAO accumulation. We assume that NAO is bound to the hydrophobic lipids and proteins in the mitochondrial membranes by hydrophobic interaction. With valinomycin, 500 ng/ml, 10 min, the mitochondria in HeLa-cells swell. Now it is possible to observe some details in the enlarged mitochondria by light microscopy. After vital staining with NAO, 5 X 10(-7) M, 10 min, the periphery of the swollen mitochondria shows an intense green fluorescence, the inner part is dark. Obviously the dye is bound to the membranes. By electron microscopy it can be shown that the valinomycin treated and NAO stained mitochondria have outer and inner membranes and cristae. They differ from untreated mitochondria mainly in the size. After incubation of the HeLa-cells with relatively high NAO concentrations, 5 X 10(-6) M, 10 min, the mitochondria show a weak orange fluorescence. It is generated by the dimers D of NAO. Therefore the dye concentration in the mitochondrial membranes is locally very high and causes dye dimerisation. The weak orange fluorescence is instable and disappears within a few seconds. Instead we observe a green fluorescence with growing intensity that is generated by the monomers M of NAO. The intensity has its maximum value after a few seconds. Using low NAO concentrations for incubation, 1 X 10(-7) M, 10 min, we observe only the green fluorescence with increasing intensity. In this case the orange fluorescence is too weak for observation (concentration quenching). It can be shown by experiments and quantum mechanics that the orange fluorescence is assigned to an optical forbidden, the green fluorescence to an allowed electronic transition of D or M respectively. Our results indicate a dissoziation of D in 2 M by irradiation of the mitochondria under the fluorescence microscope.(ABSTRACT TRUNCATED AT 400 WORDS)

[Effect of acridine dyes on the ultrastructure of mitochondria in HeLa and LM cells]. / Uber die Wirkung von Acridinfarbstoffen auf die Ultrastruktur von Mitochondrien in HeLa- und LM-Zellen.

Fluorescent staining of mitochondria in living HeLa cells and mouse fibroblasts (LM) with acridine dyes has been reported recently. The investigations are now extended to electron microscopy. Dyes used in our vital staining experiments are 10-ethyl- and 10-(2-iodethyl)acridine orange hydrochloride (EAO, IAO). The mitochondria of vitally stained cells are compared with untreated material. There are dramatic changes of the ultrastructure of HeLa mitochondria within the short incubation time of a few minutes. During the same time only small alterations of LM mitochondria could be observed.