Mineral particles in the human bronchial mucosa and lung parenchyma. II. Cigarette smokers without emphysema. The effects of cigarette smoke on the microanatomic deposition and retention pattern of exogenous mineral particles is unknown. To determine how cigarette smoke affects long-term particle retention in those with minimal smoke-related disease, we selected autopsy lungs from ten smokers without evidence of emphysema at autopsy, and used analytical electron microscopy to examine exogenous mineral particle concentration in the mucosa of seven different bronchi of varying sizes and four parenchymal sites fed by those bronchi. These data were compared with values from twelve lifetime nonsmokers. Overall, total mean particle burden in the parenchyma in the smokers and nonsmokers was similar, but there was a markedly decreased particle load retained in the smokers' airways. The consistent increase in particle burden seen in nonsmokers as airways became narrower and more distant from the carina was lost in many, but not all, of the smokers, especially for particles larger than 1 micron, and this effect did not appear to depend on amount of smoking. Rare polonium particles were found but were too sparse to use as a smoke distribution marker. However, calcium-containing particles, previously suggested by us to represent calcium carbonate actually derived from the smoke, were present in greatest concentration in the larger airways and distal parenchyma. These observations indicate the following in smokers without parenchymal smoke-induced structural damage: (1) Overall, cigarette smoking disrupts the normal retention (? deposition) pattern of particles in the airways. (2) Cigarette smoking leads to markedly decreased long-term retention of exogenous mineral particles in the airways without significantly affecting overall tissue particle retention. (3) The distribution of calcium particles in the airways and parenchyma in smokers is similar to that predicted in a recently published model of smoke particle deposition, and further supports the idea that calcium-containing particles can be used as a direct tracer of smoke particle deposition. In the present study this distribution indicates that the large airways and parenchyma receive the greatest smoke deposition. (4) Within the group of smokers, some people appear resistant to the disruptive effects of smoke on particle retention patterns, whereas other people smoking comparable amounts show markedly abnormal retention patterns. The latter may have unusual sensitivity to cigarette smoke and perhaps susceptibility to smoke-induced diseases.


Changes in the carbohydrate content of airway epithelium induced by human neutrophil elastase in the hamster. Hamster airway epithelial secretory cells were investigated by light and electron microscopic cytochemistry to study possible changes in their carbohydrate content induced by human neutrophil elastase (HNE), an agent known to cause replacement of Clara cells by mucous cells in hamster bronchi. Characterization of secretory cell carbohydrates by the AB/PAS, PA-TCH-SP, HID-TCH-SP, and LID-TCH-SP sequences indicated the existence of periodate-reactive acidic glycoconjugates, but the absence of sulfated or carboxylated glycoconjugates in both treated and control animals. Differences were seen in the quality and quantity of historeactive carbohydrates throughout various regions in the lower respiratory tract. This was especially evident in the HNE-treated animals. It is concluded that the HNE-induced expression of the mucous cell phenotype is associated with an increase in the amount of neutral and acidic nonsulfated and noncarboxylated polysaccharides stored in the secretory granules of these cells.


Inhibitory action of tetrandrine on macrophage production of interleukin-1 (IL-1)-like activity and thymocyte proliferation. Tetrandrine is a bisbenzylisoquinoline alkaloid which has been shown to exhibit antifibrotic activity against silicosis. Tetrandrine is characterized by its strong binding to alveolar macrophages and inhibition of particle-induced respiratory burst activity in these phagocytes. In contrast, tubocurine and tubocurarine are structurally similar to tetrandrine but exhibit little effect on fibrosis or activation of alveolar macrophages. The objective of the present study was to test the effect of tetrandrine on macrophage production of monokines in response to occupational dusts, and to determine tetrandrine's effect on monokine-medicated cell growth using a mouse thymocyte proliferation assay and lipopolysaccharide (LPS) as a positive control. Stimulation of alveolar macrophages by respirable silica dust resulted in a release of monokines which caused a fourfold increase in thymocyte proliferation. Coal dust, on the other hand, had no effect on macrophage production of this cytokine. Tetrandrine was found to exhibit a dose-dependent inhibition of monokine release from both silica and LPS-stimulated alveolar macrophages. In experiments where thymocytes were directly treated with tetrandrine, a dose-dependent inhibition of thymocyte proliferation was noted with both interleukin-1-(IL-1) specific and nonspecific mitogenic (concanavalin A) actions. In contrast to the inhibitory potency of tetrandrine, tubocurarine was found to have no effect on either the production of monokines by LPS-stimulated alveolar macrophages or IL-1-mediated thymocyte proliferation. These results provide a correlation between the antifibrotic effect of tetrandrine and inhibition of macrophage activation.


Monaural audiogenic seizures: evidence for control by parallel processes. SJL/J mice were binaurally sensitized and monaurally tested for susceptibility to sound-induced seizure. Most control subjects exhibited the expected biphasic pattern of running. In mice in the experimental groups, the acoustic stimulation was interrupted for less than 20 s at the conclusion of the initial burst of running. During this quiet period, the side of the blocked ear was reversed for some mice. These mice had an independent biphasic seizure when acoustic stimulation was reintroduced, indicating that the biphasic seizure progression characteristic of monaural testing is the result of a lateralized process. We conclude that the locus of this process is probably at the level of the inferior colliculus (IC).


Studies on pH regulatory mechanisms in cultured astrocytes of DBA and C57 mice. pH regulatory mechanisms in primary cultures of astrocytes from the cerebral cortex of neonatal audiogenic-seizure-susceptible DBA/2J (DBA) and genetically controlled C57BL/6J (C57) mice were studied with [14C]dimethyloxazolidine-2-4-dione (DMO) and [3H]-methyl-D-glucose (MDG). Effects of changing the concentration of Na+, K+, HCO3- or Cl- in medium, and/or of different transport blockers and metabolite inhibitor on intracellular pH (pHi) of cultured astrocytes were also studied. In nominal HCO3(-)-free HEPES-buffered Hanks' balanced salt solution (HEPES HBSS), when the pH of medium (pHo) was maintained at 7.4, the steady-state pHi of cultured astrocytes from DBA mice was 6.98 +/- 0.03, and that from C57 mice was 7.01 +/- 0.03. When the cells were incubated in HBSS containing 25 mM HCO3- and equilibrated with 5% CO2 (HCO3- HBSS, pHo = 7.4), pHi of both DBA and C57 astrocytes was approximately 0.1-0.15 pH units higher than that in HEPES HBSS. Reducing the pH or the Na+ concentration in media (pHo, [Na+]o) of either HEPES HBSS or HCO3- HBSS, pHi of both DBA and C57 astrocytes decreased markedly (0.25-0.45 pH units lower than the controls). The decrease in pHi was greater in HEPES HBSS than in HCO3- HBSS. Reducing the Cl- concentration ([Cl-]o) in either HEPES or HCO3- HBSS, pHi of astrocytes increased by 0.05-0.1 pH units. Increasing the K+ concentration ([K+]o) of or adding Ba2+ to the media increased the pHi of both DBA and C57 astrocytes accordingly. SITS, an anion transport inhibitor, decreased the pHi of both DBA and C57 astrocytes in HCO3- HBSS but not in HEPES HBSS. It enhanced the response of pHi to reduction in pHo. Amiloride, a Na(+)-H+ exchange inhibitor, decreased the pHi of both DBA and C57 astrocytes more in HEPES HBSS than in HCO3- HBSS. It enhanced the response of pHi to reduction in pHo and [Na+]o. Ouabain, an Na+,K(+)-ATPase inhibitor, decreased the pHi of cultured astrocytes in HEPES HBSS, but not in HCO3- HBSS. It also enhanced the response of pHi to changing pHo and [Na+]o in HEPES HBSS. Acetazolamide, a carbonic anhydrase inhibitor, decreased the pHi of astrocytes in both HEPES and HCO3- HBSS. Both bumetanide, an Na+,K+/Cl- cotransport blocker, and KCN, a metabolic inhibitor, produced no significant effect on the steady-state pHi or the response of pHi to changing ionic concentration in media in both DBA and C57 astrocytes.


Ontogeny of feline temporal lobe epilepsy, II: Stability of spontaneous sleep epilepsy in amygdala-kindled kittens. We previously described a model of spontaneous "sleep epilepsy" in kindled kittens with temporal lobe epilepsy (TLE). We now describe the postkindling course of this model from preadolescence to maturity and suggest pathophysiologic mechanisms. Spontaneous epilepsy, particularly generalized tonic-clonic convulsions (GTCs), developed 1h to 4 months after amygdala kindling and persisted to adulthood. At first, GTCs were detected only in sleep; later, convulsions also occurred during wakefulness. Two factors were consistently associated with the sequential onset of sleep and waking GTCs: seizure clusters and anatomic seizure localization. (1) Seizure clusters. Cats with infrequent or unclustered GTCs continued to exhibit "sleep epilepsy," defined by convulsions occurring exclusively during sleep. In contrast, cats with frequent seizure clusters developed recurrent or terminal convulsive status in conjunction with GTCs during waking and sleep. Severe seizure manifestations therefore appeared to contribute to the dissociation of convulsions from the sleep-wake cycle. (2) Anatomical seizure localization. Focal seizure origin appeared to differentiate sleep from waking GTCs. Onset during sleep was first recorded in the kindled amygdala, whereas onset during waking was initially detected outside the temporal lobe. Findings thus suggest secondary "kindling" of multifocal epilepsy. Secondary epileptogenesis is consistent with "transsynaptic" kindling effects. This phenomenon is defined in mature animals by rapid secondary site kindling (transfer) and subtle morphologic changes distal to the stimulating electrode. Transfer may be accentuated by youth, because kittens developed spontaneous seizure foci in previously unstimulated tissue. Moreover, multifocal interactions and diffuse cell loss were implicated as possible mechanisms. Collectively, the findings indicate complications with early onset TLE in kindled cats. Onset during youth can have an unfavorable prognosis, reflected by recurrent status epilepticus and multifocal epilepsy with convulsions distributed throughout the sleep-wake cycle.


Possible association of juvenile myoclonic epilepsy with HLA-DRw6. Juvenile myoclonic epilepsy (JME) is a clearly defined subform of idiopathic generalized epilepsy with a high aggregation of epilepsy in family members. With the HLA-system used as a genetic marker, a linkage between JME and the HLA region was demonstrated. Linkage with the HLA region suggests that JME may be associated with an HLA-antigen. An association could indicate that the gene lies in the HLA region and is in linkage disequilibrium with one of the HLA-antigens. Eighty-eight unrelated patients with JME were typed for the HLA-A and HLA-B locus, 77 were typed for the HLA-C locus, and 76 were typed for the DR locus. The antigen frequency was compared with those of healthy blood donors. The highest difference was noted in the frequency of DRw6 (39.5% in patients vs. 22.1% in controls). This weak association is open to question because DRw6 is known to split into DRw13 and DRw14.


Video-EEG analysis of drop seizures in myoclonic astatic epilepsy of early childhood (Doose syndrome). We studied 36 drop seizures in 5 patients with myoclonic astatic epilepsy of early childhood (MAEE) with simultaneous split-screen video recording and polygraph. Sixteen were falling attacks and 20 were either less severe attacks exhibiting only deep head nodding or seizures equivalent to drop attacks in terms of ictal pattern but recorded in the supine position. All seizures except those that occurred in patients in the supine position showed sudden momentary head dropping or collapse of the whole body downward. Recovery to the preictal position was observed in 0.3-1 s. As a result of carefully repeated observations, the 36 seizures were classified as myoclonic flexor type in 9, myoclonic atonic type in 2, and atonic type, with and without transient preceding symptoms in the remaining 25. The MF seizure was characterized by sudden forward flexion of the head and trunk as well as both arms, which caused the patient to fall. In the myoclonic atonic seizure, patients showed brief myoclonic flexor spasms, immediately followed by atonic falling. The AT seizure showed abrupt atonic falling, with and without transient preceding facial expression change and/or twitching of extremities. The ictal EEGs of all 36 seizures exhibited generalized bilaterally synchronous single or multiple spike(s) and wave discharges. Atonic drop attacks appear to be a common cause of ictal epileptic falling in MAEE.


Sleep microstructure and EEG epileptiform activity in patients with juvenile myoclonic epilepsy. Clinical and EEG manifestations of juvenile myoclonic epilepsy (JME) occur in a strict relationship to the sleep-wake cycle, particularly to transition phases (awakening, falling asleep, afternoon relaxation after work). JME manifestations are deactivated during sleep. Because arousal fluctuations during NREM sleep may be controlled by the same neurophysiologic mechanisms regulating awakening, we studied the relationship between the cyclic alternating pattern (CAP) and JME manifestations. All-night polysomnographic recordings of 10 JME patients were analyzed for variations of epileptiform EEG abnormalities in relation to sleep stages and to different microstructural variables (NCAP, CAP, phases A and B). CAP rates (ratio between total CAP duration and total NREM sleep duration) were also calculated. Average CAP rate was 46.70%, significantly higher than that (23%) of an age-matched control group. Macrostructural analysis showed only a trend toward a slight predominance of EEG epileptiform activity during slow wave sleep but no significant correlation between spiking rates and sleep stages. Microstructural analysis confirmed the CAP modulation of EEG epileptiform activity, with maximum appearance of epileptiform abnormalities during phase A CAP (normalized spiking rate = 4.00 +/- 0.98) and strong inhibition during phase B (0.06 +/- 00.6). Intermediate values were noted during NCAP (0.54 +/- 0.27). No correlation was noted between spiking rates during NREM sleep and CAP rates, possibly indicating that in JME patients the increased CAP rate may be partially independent of epileptiform EEG activity. Our data suggest that in JME patients CAP may be a neurophysiologic oscillator organizing expression of the epileptiform discharges independent of the tendency of the individual patient to produce epileptiform EEG discharges.


Late-onset epilepsy and diffuse cryptogenous cerebral atrophy. Diffuse cerebral atrophy (CA), a frequent computed tomography (CT) finding in late-onset epilepsy, is sometimes of unknown origin or cryptogenous (CCA). From a series of 228 patients with late-onset epilepsy with diffuse CA, we excluded patients with a presumed etiology. The remaining 73 (36.8%) patients were studied for a mean of 7.2 years; 15.1% had a family history of epilepsy. CCA was cortical in 50.7%, subcortical in 6.8%, and corticosubcortical in 42.5%. Seizures, generalized in 71.2%, and focal in 28.8% were generally of low occurrence with good therapeutic response. Background EEG activity was normal in most patients. During follow-up, the clinical and EEG features remained unchanged. In a control series of 92 nonepileptic subjects with diffuse CA, CCA was noted in only 4 (4.3%). No variation in grade or distribution of CCA was noted in repeated CT scans of the two groups, except in 2 epileptic patients. Psychometric tests demonstrated no cognitive impairment during evolution. The results appear to confirm the hypothesis that late-onset epilepsy associated with CCA constitutes a syndrome that is related to the characteristics of the atrophy.


Gyratory epilepsy in a patient with a thalamic neoplasm. A patient with a right thalamic oligodendroglioma developed seizures characterized by circling behavior, speech arrest, and secondarily generalized seizures. Gyratory epilepsy is relatively uncommon and may either represent a benign form of primary generalized epilepsy or occur secondary to a focal cortical lesion. Thalamic stimulation has been shown experimentally to induce circling movements, but no other clinical cases with a thalamic lesion have been described.


Nonconvulsive status epilepticus in adults: thirty-two consecutive patients from a general hospital population. We studied all adult patients who between 1984 and 1989 were initially diagnosed at our hospital as having nonconvulsive status epilepticus. Thirty-two patients fulfilled the criteria, which included ictal EEG recordings. The annual incidence was 1.5 in 100,000 inhabitants. The median age at onset of status was 51 years. Ten patients had status as their first epileptic manifestation, but most patients had a previous history of epilepsy. Median duration of epilepsy at onset of status was 4 years. Fourteen patients had focal ictal seizure activity on EEG and thus met the criteria for complex partial status. Eighteen patients had generalized seizure activity on EEG, but only 6 of these had a history of absence epilepsy or juvenile myoclonic epilepsy. None had Lennox-Gastaut syndrome. The clinical features of status in the remaining 12 patients were in some respects similar to those of the patients with complex partial status. We hypothesize that the EEG seizure activity in these patients may have been generalized from an initial focus.


Neonatal hyperekplexia: a case report.

Neonatal hyperekplexia: a case report. We report the case of a baby with transient generalized stiffness noticeable from the first days of life, hyperreflexia, massive jerks in response to sudden tactile and acoustic stimuli, and long-lasting myoclonic jerks closely resembling epileptic seizures. The father and paternal grandfather both had hyperekplexia. At age 3 years, the child had normal psychomotor development and persistent abnormal startle response to unexpected sounds or touch.


Visuomotor coordination during focal and generalized EEG discharges. A new computer-based method for continuous gauging of momentary, visuomotor coordination under simultaneous EEG control proved both effective and suitable for children. Thirty-eight patients (age 4-17 years) who had different types of hypersynchronous activity were examined: None of the patients with focal discharges showed significant impairment in the test performance. Slight losses were evident in less than 3-s generalized, irregular spike-wave paroxysms. Deterioration was particularly evident in the bilateral synchronous 3-Hz spike-wave series, which lasted greater than 3 s.


Placebo-controlled pilot study of centromedian thalamic stimulation in treatment of intractable seizures. Stimulation of centromedian (CM) thalamic nuclei has been proposed as a treatment for seizures. We implanted programmable subcutaneous (s.c.) stimulators into CM bilaterally in 7 patients with intractable epilepsy to test feasibility and safety. Stimulation was on or off in 3-month blocks, with a 3-month washout period in a double-blind, cross-over protocol. Stimuli were delivered as 90-microseconds pulses at 65 pulses/s, 1 min of each 5 min for 2 h/day, with voltage set to half the sensory threshold. Stimulation was safe and well-tolerated, with a mean reduction of tonic-clonic seizure frequency of 30% with respect to baseline when stimulator was on versus a decrease of 8% when the stimulator was off. There was no improvement in total number of generalized seizures with stimulation, and treatment differences were not statistically significant. Stimulation at low intensity did not alter the EEG acutely, but high-intensity stimulation induced slow waves or 2-3 Hz spike-waves with ipsilateral frontal maximum. In an open-label follow-up segment with stimulator trains continuing for 24 h/day, 3 of 6 patients reported at least a 50% decrease in seizure frequency. There were no side effects. This pilot project demonstrated the feasibility of controlled study of thalamic stimulation in epilepsy, but further study will be needed to demonstrate efficacy.


Temporal neocorticectomy in management of intractable epilepsy: long-term outcome and predictive factors. We report the results of a long-term follow-up study of 50 patients who underwent removal of temporal neocortex with preservation of deeper limbic structures as surgical therapy for intractable temporal lobe epilepsy. The follow-up period ranged from 3 to 15 years. Preoperative EEG investigations were based on interictal discharges alone. Three factors were predictive of a good outcome: (a) A clear unilateral anterior-midtemporal focus (p less than 0.01), (b) stereotypical onset of temporal lobe seizure (p less than 0.005), and (c) greater volume of tissue removed at operation (p less than 0.05). Overall results showed that 62% of patients experienced an outcome of "cure" or "almost cure," as classified according to a modified version of Crandall's criteria (Crandall's I and II). Those who experienced a significant reduction in seizures but who continued to have intractable epilepsy (Crandall's III) were not considered to have had a good result. Overall outcome compares favorably with other that of centers using different surgical approaches and indicates that neocorticectomy is a suitable procedure in a highly selected population even when limited resources are available.


Anatomic correlates of interhippocampal seizure propagation time. The relation between interhippocampal seizure propagation time (IHSPT) and anatomic alterations in the human epileptic hippocampus may provide insight into the pathophysiology of temporal lobe epilepsy (TLE). Using depth electrode recordings, we measured the time required for spontaneous seizures with onset in one hippocampus to become manifest in the contralateral hippocampus in 50 patients who underwent resection of the temporal lobe of seizure origin. Cell densities in individual hippocampal subfields were determined and correlated with mean IHSPT for each patient. Mean IHSPT was significantly and inversely correlated with cell counts in CA4 only (r = -0.38, p less than 0.01, Pearson's product correlation; r = -0.52, p less than 0.001, Spearman's rank order correlation). In 5 patients with bilateral independent hippocampal seizure onset who had temporal lobectomy and a diagnosis of mesial temporal sclerosis, mean IHSPT was consistently longer from the sclerotic temporal lobe than to it. These observations suggest that anatomic changes associated with chronic epilepsy alter propagation patterns. Because CA4 is believed to modulate the output of dentate granule cells and also has commissural connections to the contralateral homotopic area, the association of decreased CA4 cells with prolongation of IHSPT suggests that the observed anatomic alterations may actively (through increased inhibition) or passively (through decreased recruitment) interfere with various routes of seizure propagation.


Ictal and interictal activity in partial epilepsy recorded with multichannel magnetoelectroencephalography: correlation of electroencephalography/electrocorticography, magnetic resonance imaging, single photon emission computed tomography, and positron emission tomography findings. Ictal and interictal epileptic activity was recorded for the first time by multichannel magnetoencephalography (MEG) in three patients with partial epilepsy. Pre- and intra-operative localization of the epileptogenic region was compared. The interictal epileptic activity was localized at the same region of the temporal or frontal lobe as the ictal activity. Main zones of ictal activity were shown to evolve from the tissue at the centers of interictal activity. Pre- and intra-operative electrocorticography (ECoG) as well as postoperative outcome confirmed localization in the temporal and frontal lobe. Results also correlated with findings from scalp EEG, interictal and ictal single photon emission computed tomography (SPECT), positron emission tomography (PET), and magnetic resonance imaging (MRI). Combined multichannel MEG/EEG recording permitted dipole localization of interictal and ictal activity.


Magnetic resonance imaging in localization of EEG depth electrodes for seizure monitoring. Preoperative evaluation of patients with medically refractory complex partial seizures requires intensive EEG monitoring. Part of this process may include insertion of depth electrodes into temporal lobe structures. Although many imaging methods have been used for stereotactic placement of the electrodes, only computed tomography (CT) has been used to verify position after implantation. We studied 30 patients in whom magnetic resonance imaging (MRI) was used to visualize the position of stainless-steel electrodes stereotactically implanted in the temporal lobes. The procedure is noninvasive, does not utilize radiation, and has the additional advantages of multiplanar capability, increased soft tissue contrast, and lack of bony artifacts commonly associated with CT.


Seizures during cognitive testing in patients with temporal lobe epilepsy: possibility of seizure induction by cognitive activation. In 18 of 185 patients under consideration for epilepsy surgery, 20 seizures were observed during neuropsychological evaluation. We wished to determine whether the task at seizure onset corresponded neuropsychologically to lateralization of the epileptic focus. The patients' characteristics and the circumstances of the seizures were as follows: Fourteen patients had right temporal lobe epilepsy (RTE) and four had left temporal lobe epilepsy (LTE). Although a wide range of cognitive functions had been tested, all but one seizure occurred during assessment of memory performance. In the RTE patients, 12 of 16 seizures occurred during visual memory testing. Two seizures were observed during a verbal memory task, and one seizure was observed during mental rotation. In two LTE patients, seizures were elicited during verbal memory testing. Two LTE patients with seizures during visual memory testing had speech dominance of the right hemisphere. This high correspondence between the eliciting performance and the focus localization suggests that cognitive performances ipsilateral to the epileptic focus may affect seizure threshold in focal epilepsies.


Psychosocial difficulties and outcome after temporal lobectomy. One hundred fifteen consecutive patients in the Austin Hospital Comprehensive Epilepsy Program (Melbourne, Australia) were surveyed to document the psychosocial and rehabilitation difficulties after temporal lobectomy. During the follow-up period (mean 4 years) 3 patients died, 5 patients were lost to follow-up, and 107 patients with family and friends were interviewed. Eighty-four patients (78%) had been seizure-free for the year preceding the interview; 13 others had seizure reduction greater than 75%. Success in ablation or reduction in seizures correlated with the amount of postoperative gain, but in this series, analysis of work and dependency outcome did not emphasize areas of success. Although improvement in work and financial status, interpersonal relations and sexuality were all recorded, successful patients deemed that most advance had been made in the areas of newly acquired independence, enhanced career potential, and social freedom. Significant postoperative anxiety, especially after left temporal lobectomy, was noted, possibly explained by benzodiazepine antiepileptic drug (AED) discontinuation. Although 1 patient committed suicide, neither depression nor psychosis was common in the rehabilitation period, in contrast to results in previous series. Significant sociodomestic problems emerged from this survey, however: 35% of patients considered successes reported postoperative problems stemming from the necessity to restructure family dynamics; in 6%, this resulted in divorce. Moreover, 20% of patients and relatives reported significant behavioral problems in coping with the seizure-free lifestyle. Finally, the problems of the worsened situation after surgical failure indicated the counterproductive potential of ineffective lobectomy. These results indicate the necessity for a preoperative counseling program to prevent these problems.


Antiepileptic drug treatment after temporal lobe epilepsy surgery: a randomized study comparing carbamazepine and polytherapy. Temporal lobectomy is an effective treatment in selected patients with medically intractable temporal lobe epilepsy (TLE). Postoperative antiepileptic drug (AED) treatment guidelines have not been established, and patients are often treated with polytherapy postoperatively. We prospectively randomized 40 patients undergoing temporal lobectomy to monotherapy with carbamazepine (CBZ, 20) or to continuation of their presurgical polytherapy (20) to assess the efficacy and safety of each regimen during the first year after operation. No significant differences between groups were noted with respect to seizure recurrence rate and type or time of recurrence. Patients in the polytherapy group had a 30% incidence of drug-related side effects as compared with only 10% in the CBZ group. These results suggest that after temporal lobectomy for intractable epilepsy, patients can be safely treated with CBZ monotherapy and that treatment with multiple AEDs is not necessary.


Lidocaine in refractory status epilepticus: confirmation of efficacy with continuous EEG monitoring. Lidocaine was efficacious in 2 patients with refractory status epilepticus (RSE) unresponsive to several antiepileptic drugs (AEDs), including high-dose barbiturates. We confirmed the efficacy of lidocaine with, for the first time in adults, continuous EEG monitoring. Lidocaine, when properly used, may be a treatment option in RSE.


Gamma-vinyl GABA (vigabatrin) in epilepsy: clinical, neurochemical, and neurophysiologic monitoring in epileptic patients. We report long-term clinical, neurochemical, and electrophysiologic data of gamma-vinyl GABA (GVG, vigabatrin) in three groups of patients. GVG was started as add-on therapy for 75 patients with refractory complex partial seizures (group A) and for 36 mentally handicapped patients with severe epilepsy (group B). The third group (C) consisted of 20 patients with carbamazepine (CBZ) monotherapy, in half of whom GVG monotherapy was substituted. After 3 months, 55% of patients in group A and 42% in group B were responders (reduction in seizure frequency greater than 50%). After 6 (group A) and 3 years (group B) of follow-up, 27 and 33% of the patients, respectively, still had good response to GVG. Neurochemical measurements showed a twofold increase in CSF GABA concentrations and minimal or no changes in other neurotransmitter-related parameters. In group C, substitution of GVG as medication tended to normalize the lengthened latencies in somatosensory evoked potentials (SEPs) observed during CBZ treatment.


Decreased sialidase activity in alveolar macrophages of guinea pigs exposed to coal mine dust. The origin of immune dysfunctions that are observed in pneumoconiotic miners still remains unknown. There is evidence that the carbohydrate moiety of membrane glycoconjugates is of primary importance in many functions of immunocompetent cells. The glycosylation, and especially the sialylation level of membrane components of various lymphocyte and macrophage subsets, vary depending on the state of cellular differentiation and activation. Sialidases, which may regulate the amount of sialic acids exposed on the cell membrane, can thus be considered as immunoregulatory enzymes. In this report, the sialidase activity has been measured in alveolar macrophages (AM) and in cell-free bronchoalveolar lavage fluid (BALF) from guinea pigs exposed for 4 months to coal mine dust at a concentration of 300 mg/m3. The samples were collected by bronchoalveolar lavage 2 months after cessation of exposure. The sialidase activity in the cell-free fluid and in the purified alveolar macrophages showed a 10-fold decrease (p less than 0.001). Kinetic parameters of the enzyme such as Km and optimum pH did not change. This changed activity was specific for sialidase, as two other lysosomal glycosidases, beta-galactosidase and N-acetylglucosaminidase, showed unchanged activities. These results suggest the possibility that, by inducing a decreased sialidase activity, exposure to coal mine dust may lead to a modified expression of AM membrane-associated sialic acids giving rise to altered immune functions (i. e., phagocytosis, antigen processing, response to cytokines, etc.).


Gamma-vinyl GABA (vigabatrin): relationship between dosage, plasma concentrations, platelet GABA-transaminase inhibition, and seizure reduction in epileptic children. The relationship between vigabatrin gamma-vinyl GABA (GVG, vigabatrin) daily dosage or steady-state plasma concentrations (CSS), platelet GABA-transaminase (GABA-T) inhibition, and seizure reduction were studied in 16 children with refractory epilepsy. After 2 months of observation and 1 month of single-blind add-on placebo, a fixed GVG dosage was added for 2 months. The dosage was then adjusted in two 2-month periods each, based on the patient's clinical response. In the fixed-dose period, GVG dosages of 56.8 mg/kg/day and CSS of 8.1 mg/L reduced GABA-T activity from 13.9 to 5.1 pmol/min/mg protein (p less than 0.001) and that of seizures from 51.4 to 22.3 seizures per month (p less than 0.01). Seizure reduction was correlated with dosage (r = 0.83, p less than 0.001), but not with CSS or with platelet GABA-T inhibition. After the GVG dose-adjustment periods, in which dosages of 84.4 mg/kg/day and CSS of 10.6 mg/L were reached, only a slight reduction was observed in both GABA-T activity (from 5.1 to 4.9 pmol/min/mg protein) and seizures (from 22.3 to 18.1 seizures per month). In GVG-responsive patients (excluding placebo-sensitive and GVG-resistant patients), a greater reduction of seizures was achieved (from 17.0 to 7.1 seizures per month, p less than 0.05), which was not accompanied by greater inhibition of GABA-T. GVG treatment in children should be started with a dosage of 50 mg/kg/day, increased to 75 or even 100 mg/kg/day when a partial response is observed. If seizures do not improve or if they become worse, the patient should be considered resistant and GVG should be discontinued.


Total cholesterol, high-density lipoprotein cholesterol, and triglycerides in children receiving antiepileptic drugs. The influence of antiepileptic drug (AED) therapy on total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, and triglycerides was studied in 208 epileptic children compared with 175 normal children. A significant increase in TC plasma levels was observed with carbamazepine (CBZ), phenobarbital (PB), and phenytoin (PHT). The patients receiving valproate (VPA) showed levels very similar to those of the control population. The results may be explainable by the different biotransformation pathway of these drugs. HDL cholesterol and triglycerides were not altered by any of the AEDs. We recommend monitoring TC level in patients receiving CBZ, PB, and PHT and prescription of diet treatment, at least during the time of AED treatment.


Factors affecting the in vitro dissolution of cobalt oxide. In a recent interspecies comparison of the lung clearance of cobalt oxide (57Co3O4), differences of up to 4-fold were found in the translocation rates of 57Co to blood between seven different animal species, including man. This study investigated some factors that could influence the dissolution of this material in vitro. The effect of bicarbonate and citrate concentrations (over physiological ranges) and medium pH on in vitro dissolution of 57Co from 57Co3O4 particles was measured in a simple noncellular system. pH levels of 4.5, 6.1, and 7.2 were used to correspond to those in the alveolar macrophage lysosome, its cytoplasm, and the extracellular lung fluid. Measurements of the fractional dissolution rate were made weekly for 3 months. pH had the greatest effect on dissolution rates, with particles suspended in the lowest pH medium (4.5) dissolving at a significantly faster rate than at higher pH values. Increasing citrate concentrations resulted in slightly higher dissolution rates, but there was no effect of bicarbonate concentration. There was no evidence of synergism between the factors studied.


Electrochemistry of anticonvulsants: electron transfer as a possible mode of action. Reduction potentials were determined for various anticonvulsants, including progabide, SL 75.102, CGS 9896, pyridazines, zonisamide, 1,2,3-triazoles, and copper complexes. The values generally were in the range of about -0.1 to -0.6 V for the protonated drugs and the metal complexes. Reduction potentials provide information on the feasibility of electron transfer (ET) in vivo. If the value is relatively positive (greater than about -0.6 V), the agent can act catalytically as an electron acceptor from an appropriate cellular donor. A concomitant favorable influence on abnormal neuronal processes associated with epilepsy could occur. We describe ET as a possible mode of action of anticonvulsants as well as some antiepileptic agents with no electrochemical data based on this hypothetical ET approach.


Recovery of rat alveolar macrophages by bronchoalveolar lavage under normal and activated conditions. When rat (female Wistar) lungs were lavaged (bronchoalveolar lavage, BAL) six times with physiological saline, approximately the same number of alveolar macrophages (AM) were found in the first and second BAL, whereas in the third fourth, fifth, and sixth BAL, the number of AM decreased exponentially. Morphometric counting of the number of AM in histological sections of lung tissue showed that only 14% of the AM population had been recovered by BAL. Although additives to the BAL fluid such as lidocaine and/or fetal calf serum increased the AM count in the first washing considerably, the total number of AM washed out remained unaltered. Addition of the phagocytosis stimulant zymosan increased the AM count in BAL by a factor of more than 2. On stimulation of the lungs with an inert dust (silicon carbide), the AM count in the BAL and the lung was only slightly increased 8 weeks after intratracheal instillation. In contrast, after exposure to fibrogenic and cytotoxic quartz, the AM count in BAL and lung was significantly increased, and the recovery of AM had also increased by a factor of approximately 2. The experiments show that it is the micromilieu of the alveoli and the condition of the AM (certain physiological activation states, such as phagocytic activity) that essentially determine the degree of recovery.


Intraphagolysosomal pH in canine and rat alveolar macrophages: flow cytometric measurements. Intracellular dissolution of inhaled inorganic particles is an important clearance mechanism of the lung and occurs in phagolysosomal vacuoles of phagocytes. Flow cytometric measurements of intraphagolysosomal pH in alveolar macrophages (AM) obtained from beagle dogs, Wistar rats, and from a baboon were made using fluorescein isothiocyanate-labeled amorphous silica particles (FSP). AM were obtained by bronchoalveolar lavage. FSP were phagocytized by AM in cell suspensions incubated in full media for 24 hr up to 6 days. Dual laser flow cytometry was performed and six-parameter list mode data were recorded from forward scatter, side scatter, and fluorescence intensities at 530 nm excited at 457 nm and 488 nm as well as logarithmic fluorescence intensity at wavelengths 630 nm excited at 488 nm. In this way it was possible to discriminate viable AM with phagocytized FSP from lysing AM with phagocytized FSP and from cells without FSP and from free FSP. Viable cells were distinguished from lysing cells by staining with propidium iodide immediately before the flow cytometric measurement. A calibration curve for the pH value was determined from FSP suspended in buffered media at pH values ranging from 3.5 to 7.5. First flow cytometrical results indicated that after an incubation time of 24 hr, the mean intraphagolysosomal pH of viable AM was 4.7 +/- 0.3 for dogs and 5.1 +/- 0.5 for rats. The intraphagolysosomal pH of the baboon AM was 4.5.


Intracellular particle dissolution in alveolar macrophages. Aerosol particles deposited in the lungs that are not readily soluble in the epithelial lining fluid will be phagocytized by alveolar macrophages (AM). Inside the phagolysosomal vacuole, the constituents of the plasma allow dissolution of a variety of compounds at a higher rate than dissolution in extracellular lung fluids. Chelator concentration and a pH value of about 5 were found to control intracellular particle dissolution (IPD). Hence, IPD is the initial step of translocation of dissolved material to blood, which is an important lung clearance mechanism for particles retained long term. IPD rates of uniform test particles determined in human, baboon, and canine AM cultures were similar to initial translocation rates determined in lung clearance studies of the same species after inhalation of the same test particles. IPD rate in cultured AM proved to be a sensitive functional parameter of AM, which was used to identify changes in the clearance mechanism of translocation during different exposure conditions.


In vitro dissolution of uranium oxide by baboon alveolar macrophages. In vitro cellular dissolution tests for insoluble forms of uranium oxide are technically difficult with conventional methodology using adherent alveolar macrophages. The limited number of cells per flask and the slow dissolution rate in a large volume of nutritive medium are obvious restricting factors. Macrophages in suspension cannot be substituted because they represent different and poorly reproducible functional subtypes with regard to activation and enzyme secretion. Preliminary results on the dissolution of uranium oxide using immobilized alveolar macrophages are promising because large numbers of highly functional macrophages can be cultured in a limited volume. Cells were obtained by bronchoalveolar lavages performed on baboons (Papio papio) and then immobilized after the phagocytosis of uranium octoxide (U3O8) particles in alginate beads linked with Ca2+. The dissolution rate expressed as percentage of initial uranium content in cells was 0.039 +/- 0.016%/day for particles with a count median geometric diameter of 3.84 microns(sigma g = 1.84). A 2-fold increase in the dissolution rate was observed when the same number of particles was immobilized without macrophages. These results, obtained in vitro, suggest that the U3O8 preparation investigated should be assigned to inhalation class Y as recommended by the International Commission on Radiological Protection. Future experiments are intended to clarify this preliminary work and to examine the dissolution characteristics of other particles such as uranium dioxide. It is recommended that the dissolution rate should be measured over an interval of 3 weeks, which is compatible with the survival time of immobilized cells in culture and may reveal transformation states occurring with aging of the particles.


In vitro dissolution of curium oxide using a phagolysosomal simulant solvent system. Detailed study of actinide oxide behavior in alveolar macrophages (AM) in vitro is limited because of the short life span of these cells in culture. We created an in vitro dissolution system that could mimic the acidic phagolysosomal environment for the actinide and be maintained for an indefinite period so that dissolution of more insoluble materials could be measured. The dissolution system for this investigation, consisting of nine different solutions of HCl and the chelating agent diethylenetriamine pentaacetate (DTPA) in distilled water, is called the phagolysosomal simulant solvent (PSS). In this system, both the pH and the amount of DTPA were varied. We could observe the effect of altering pH within a range of 4.0-6.0 (similar to that of the phagolysosome) and the effect of the molar ratio of DTPA to curium at 1000:1, 100:1, or 10:1. We chose curium sequioxide (244Cm2O3) to validate the PSS for actinide dissolution versus that occurring in AM in vitro because it dissolves significantly in less than 1 week. The polydisperse 244Cm2O3) aerosol was generated, collected on filters, resuspended, and added to the PSS solutions and to cultured canine AM. By comparing dissolution in the two systems directly, we hoped to arrive at an optimum PSS for future dissolution studies. PSS and cell culture samples were taken daily for 7 days after exposure and tested for the solubilized curium. The amount of soluble material was determined by ultracentrifugation to separate the insoluble Cm2O3 from the soluble curium in the PSS solutions and filtration for the cell-containing material.(ABSTRACT TRUNCATED AT 250 WORDS)


Blood-brain barrier penetration of felbamate. The brain uptake index (BUI) method of Oldendorf was used to examine blood-brain barrier (BBB) drug transport in mice, rats, and rabbits; felbamate (FBM) extraction (E) in a single transcapillary passage was 5-20%, and drug uptake in rat brain was not concentration-dependent. Like diazepam, FBM was retained in mouse brain. To ensure that radioactivity measurements reflected the disposition of parent drug and not some metabolite, extracts of mouse brain were prepared for further analysis. No FBM metabolites were detected in brain 5 min after administration: In silica gel thin-layer chromatography (TLC), a single [14C]FBM peak was detected--Rf = 0.504 (70:30 acetone:hexane). Confirmatory high-performance liquid chromatography (HPLC) separations [30% methanol, 1.3 ml/min, C18 column, ultraviolet (UV) detection 254 nm] indicated a single peak containing greater than 93% of the radioactivity in the FBM fraction (12-min retention time). In a single transit through the liver (a nonbarrier tissue with fenestrated capillaries), FBM E was 82%. The octanol:buffered saline partition coefficient of FBM was (log PFBM =) 0.54 +/- 0.01. Thus, lipid-mediated BBB penetration of FBM is similar to that of phenytoin (PHT) and phenobarbital (PB). Plasma proteins do not affect FBM entry to the brain: neither human serum, nor bovine or human serum albumin (BSA, HSA), nor human alpha 1 acid glycoprotein (orosomucoid) significantly modified BBB FBM extraction. Erythrocyte-borne FBM may also dissociate and gain access to the brain in a single transcapillary passage. Differences between newborn and adult rabbit BBB FBM extraction and between different anesthetic agents are attributable to cerebral blood flow (CBF) rates. The permeability-surface area products (PS = [CBF].[E]) for FBM in rats, rabbits, and mice were 0.09, 0.16 and 0.30 ml/min/g, respectively. Preliminary autoradiographic analyses of frozen brain sections suggest that [14C]FBM distributes relatively uniformly throughout the brain and that minor variations apparently are a function of differing CBF rates.


In vitro chemical and cellular tests applied to uranium trioxide with different hydration states. A simple and rapid in vitro chemical solubility test applicable to industrial uranium trioxide (UO3) was developed together with two in vitro cellular tests using rat alveolar macrophages maintained either in gas phase or in alginate beads at 37 degrees C. Industrial UO3 was characterized by particle size, X-ray, and IR spectra, and chemical transformation (e.g., aging and hydration of the dust) was also studied. Solvents used for the in vitro chemical solubility study included carbonates, citrates, phosphates, water, Eagle's basal medium, and Gamble's solution (simulated lung fluid), alone, with oxygen, or with superoxide ions. Results, expressed in terms of the half-time of dissolution, according to International Commission on Radiological Protection (ICRP) classification (D,W,Y), varied for different hydration states of UO3, showing a lower solubility of hydrated UO3 in solvents compared to basic UO3 or UO3 heated at 450 degrees C. Two in vitro cellular tests on cultured rat alveolar macrophages (cells maintained in gas phase and cells immobilized in alginate beads) were used on the same UO3 samples and generally showed a lower solution transfer rate in the presence of macrophages than in the culture medium alone. The results of in vitro chemical and cellular tests were compared, with four main conclusions: a good reproducibility of the three tests in Eagle's basal medium the effect of hydration state on solubility, the classification of UO3 in terms of ICRP solubility criteria, and the ability of macrophages to decrease uranium solubility in medium.


Pharmacokinetics of felbamate in pediatric and adult beagle dogs. The relative bioavailability and pharmacokinetics of felbamate (FBM) after a single oral dose and after 10 once-daily oral doses of 60 mg/kg were investigated in adult and pediatric dogs of both sexes. The pediatric and adult dogs were aged 4-6 weeks and 1-2 years, respectively. Analysis of variance (ANOVA) was performed on the bioavailability parameters among all groups and between the first and last doses. No sex-related differences in bioavailability and pharmacokinetic parameters were observed. The bioavailability of FBM in pediatric dogs was significantly less as compared with that in adult dogs. Rapid overall elimination of the drug in pediatric dogs appears to be responsible for the lower bioavailability. The bioavailability of FBM after the last dose was also significantly lower than after the first dose for both age groups. No major differences in the rate constant of FBM absorption (ka) and volume of distribution at steady state (VSS) were observed between the two age groups. As with other clinically useful antiepileptic drugs (AEDs), higher doses of FBM may be required in pediatric populations to achieve optimum drug levels, assuming that age-related changes in FBM disposition will also be confirmed in humans.


Role of alveolar macrophages in precipitation of mineral elements inhaled as soluble aerosols. The lysosomes of several varieties of cells such as the tubular proximal cell of the kidney and the alveolar macrophage have the ability to concentrate and precipitate several elements inhaled in water-soluble form, usually as phosphate. The mechanism involved is attributed to the high acid phosphatase activity of lysosomes and can be considered as an in vivo Gomori reaction. Among the elements studied, most of them are chemotoxic or radiotoxic (Cr; group IIIA: Al, Ga, In; rare earths: La, Ce, Tm; actinides: Th, U). In the lung macrophage, this mechanism of intralysosomal concentration and precipitation may prevent the diffusion of these toxic elements through the alveolar membrane.


Phagolysosomal pH and dissolution of cobalt oxide particles by alveolar macrophages. We studied phagolysosomal pH in rabbit alveolar macrophages (AM) incubated with 0-15 microM chloroquine. There was a dose-related increase in pH with chloroquine concentration. Electron microscopy showed that chloroquine increased lysosomal size. In a second experiment we studied dissolution of radiolabeled cobalt oxide particles by rabbit AM, phagolysosomal pH, and lysosomal size. The cells were incubated for 2 days with 0, 2.5, and 10 microM chloroquine. Size and pH increased with chloroquine concentration. Dissolution of cobalt particles by the AM did not clearly change with pH. In a third experiment, dissolution of cobalt oxide particles in 0.1 M acetate buffer in saline with pH 4.0, 5.0, and 6.0 was studied. At the same pH, dissolution in acetate buffer was faster than in the AM, and the dissolution appeared to decrease faster with increasing pH than in the AM. A simple model for dissolution of a particle in a phagolysosome was proposed. This model predicts the types of differences in dissolution between AM and buffered saline.


The effect of gamma-irradiation on the adherent capacity and iron metabolism of alveolar macrophages in mice and rats. The effect of external gamma-irradiation on the survival of alveolar macrophages (AM) and on the extracellular release of 59Fe from AM-ingested [59Fe]iron hydroxide colloid was investigated in vitro using cells from C3H mice and Wistar rats. 59Fe release from mouse AM was enhanced by irradiation in a dose-dependent fashion, but irradiation up to 112 Gy did not affect the release from rat AM. Cell survival of AM, measured by direct counting of cell nuclei of adherent AM or vital staining with crystal violet, decreased dose dependently in mice and rats, but mouse AM were more radiosensitive than rat AM.


Effect of 60Co gamma-irradiation on the nonspecific cytotoxicity of alveolar macrophages in vitro. This paper reports on the effect of radiation on the nonspecific cytotoxicity of rat alveolar macrophages (AM). AM (effector cells) of bacille Calmette-Guerin-activated Wistar rats were irradiated with 60Co gamma rays in vitro to give doses of 0, 100, 300, and 500 Gy. Three hours after irradiation, the AM were cultured with human lung adenocarcinoma AGZY83-a and HeLa target cells in 125I-deoxyuridine-containing media for 6 hr and the cytotoxicity indexes determined. The results indicated that the cytotoxicity indexes of AM against human lung adenocarcinoma cells and HeLa cells were 94.3 +/- 0.3% and 81.3 +/- 1.9%, respectively. The cytotoxicity indexes using an effector/target cell ratio (E/T) of 10 seemed to be greater than with ratios of 20 and 30. The cytotoxicity indexes of AM (7 rats), irradiated to give doses of 0, 100, 300, and 500 Gy, against adenocarcinoma cells at an E/T ratio of 10 were 87.9 +/- 8.4%, 65.4 +/- 14.1%, 47.5 +/- 17.5%, and 36.7 +/- 9.7%, respectively. The significance of the nonspecific cytotoxicity of AM in the immunological elimination of tumors and the inhibitory effect of radiation on AM cytotoxicity are discussed.


Activity testing of alveolar macrophages and changes in surfactant phospholipids after irradiation in bronchoalveolar lavage: experimental and clinical data. This study presents results of bronchoalveolar lavage (BAL) after irradiation to the lungs in mice as well as clinical data. The number of BAL cells, mainly macrophages, lymphocytes, and granulocytes, changed in a time-dependent manner. The phagocytic activity of the macrophages measured as the phagocytosis of microbeads and measured as the esterase activity also showed a strong time-dependent increase during the acute phase up to 21 days after irradiation. The contents of surfactant phospholipids (SF) and sphingomyelin (SPH; as a parameter for cell death) were quantified by HPLC. Both were significantly changed between day 2 and 21 after irradiation. Three BALs of a patient with idiopathic interstitial pneumonitis, who had received an allogenic bone marrow graft after total body irradiation with 10 Gy, showed similar effects in the cellular and surfactant parameters. These data indicate that there are positive interactions between the number of different BAL cells, macrophage activity, and SF and SPH content in the preclinical model of the mouse as well as in the clinical situation after lung irradiation.


Pulmonary and thoracic macrophage subpopulations and clearance of particles from the lung. Pulmonary macrophages consist of several subpopulations that can be defined by their anatomical locations as well as by other criteria. In addition to the well-known alveolar macrophages that reside on the alveolar surface, pulmonary macrophages also occur in the conducting airways, in various pulmonary interstitial regions, and, in some mammalian species, in the lung's intravascular compartment. Other thoracic macrophages of relevance to pulmonary defense and some lung disease processes are the pleural macrophages resident in the pleural space and macrophages present in regional lymph nodes that receive lymphatic drainage from the lung. Of the above subpopulations of pulmonary and thoracic macrophages, the alveolar macrophages have received the most experimental attention in the context of the pulmonary clearance and retention of deposited particles. Accordingly, less information is currently available regarding the roles other pulmonary and thoracic populations of macrophages may play in the removal of particles from the lower respiratory tract and associated tissue compartments. This report provides an overview of the various subpopulations of pulmonary and thoracic macrophages, as defined by their anatomical locations. The known and postulated roles of macrophages in the pulmonary clearance and retention of particles are reviewed, with particular emphasis on macrophage-associated processes involved in the pulmonary clearance of relatively insoluble particles.


Reaction of alveolar macrophages to inhaled metal aerosols. For more than a decade we have exposed rabbits to different metals, usually in soluble form, and investigated the effects on the lungs. The metal concentrations have been around 1 mg/m3,i.e., not more than a factor of 10 above occupational threshold limit values. The exposure periods have been 1-8 months (6 hr/day, 5 days/week). We have studied especially the morphology and function of alveolar macrophages (AM), the morphology of alveolar type I and type II epithelial cells, and analyzed lung phospholipids. Several metals produce specific, complex effects. For example, metallic and soluble nickel (NiCl2) increase both number and size of the type II cells, increase the production of surfactant, and affect morphology and function of AM. Cobalt (CoCl2) induces a different effect on type II cells from nickel, causing the formation of nodules in these cells. Trivalent chromium [Cr(NO3)3] does not affect either type II cells or the amount of surfactant significantly, but markedly affects AM. The administered metals affect AM both directly and indirectly. For example, nickel induces an increased production of surfactant, resulting in overfed AM with an increased metabolic activity. However, nickel also induces a direct decrease in the release of lysozyme activity by AM. Our results emphasize the complexity of the effects on the lungs of inhaled agents, which can act both directly and indirectly on AM.


Irreversible pulmonary changes induced in rat lung by dust overload. The objective of this study was to investigate whether the effects of dust overload are reversible upon cessation of subchronic exposure to test toner. Female rats were exposed 6 hr/day, 5 days/week for 3 months to a test toner at 0, 10, and 40 mg/m3. The retained quantity of test toner in the lungs at the end of exposure was 0.4 and 3.0 mg for the low and high exposure groups, respectively. Fifteen months later, the corresponding values were 0.12 and 2.65 mg in the lungs. Alveolar clearance of tracer aerosols as well as cytologic and enzymatic parameters in the bronchoalveolar fluid was investigated at the end of exposure and subsequently up to 15 months later. The alveolar clearance of 59Fe2O3, 51Cr-polystyrene, and 85Sr-polystyrene tracer aerosols was slightly retarded at the low and substantially impaired at the high exposure level. At the low exposure level, there was some recovery in the clearance behavior up to 6 months after exposure. In contrast, at the high exposure level there was no indication of a reversal of the impaired clearance. For the beta-glucuronidase activity and the number of polymorphonuclear cells, the pattern of the effects was similar to the effects on the half-time tracer particle clearance. In conclusion, the dust overload at a lung burden of 3 mg test toner in rats was persistent for at least 15 months after termination of exposure.


Role of the alveolar macrophage in lung injury: studies with ultrafine particles. We conducted a series of experiments with ultrafine particles (approximately 20 nm) and larger particles (less than 200 nm) of "nuisance" dusts to evaluate the involvement of alveolar macrophages (AM) in particle-induced lung injury and particle translocation in rats. After intratracheal instillation of both ultrafine particles and larger particles of TiO2, we found a highly increased interstitial access of the ultrafine particles combined with a large acute inflammatory reaction as determined by lung lavage parameters. An additional experiment revealed that intratracheal instillation of phagocytized ultrafine TiO2 particles (inside AM) prevented both the pulmonary inflammatory reaction and the interstitial access of the ultrafine particles. Another experiment showed that the influx of polymorphonuclear cells (PMN) into the alveolar space unexpectedly decreased with higher doses of ultrafine particles, whereas alveolar epithelial permeability (protein leakage) increased. The divergence between PMN influx into the alveolar space and changes in alveolar epithelial permeability implies that they are separate events. Pulmonary inflammatory parameters determined by lung lavage analysis correlated best with the surface area of the retained particles rather than with their mass, volume, or numbers. Because higher doses resulted in an increased interstitialized fraction of particles, we suggest that inflammatory events induced by particles in the interstitial space can modify the inflammation in the alveolar space detectable by lung lavage. Our results demonstrate the dual role of AM for modifying particle-induced lung injury, i.e., both preventing such injury and contributing to it. We conclude that the increased pulmonary toxicity of ultrafine particles is related to their larger surface area and to their increased interstitial access.(ABSTRACT TRUNCATED AT 250 WORDS)


Dosimetric implications of pulmonary macrophage clusters observed within lungs of rats that have inhaled enriched UO2 particles. Twenty-four Fischer 344 rats were exposed to enriched uranium dioxide (UO2) aerosols to give a mean initial lung burden of 291 +/- 89 (SD) micrograms. Groups of rats were killed at 1, 7, 180, 360, 540, and 720 days post-inhalation (PI). Their lungs were fixed and inflated. Sections cut from all five lung lobes were used to prepare CR-39 neutron-induced 235U fission fragment autoradiographs. A single traverse across a CR-39 autoradiograph of a tissue section, from the left lung of all the rats, was made using a motorized microscopic stage. The traverse was divided into 10 fields. The track counts per field were used to test for homogeneity of track distribution and to assess if there was any tendency for tracks to be related to the peripheral region of the lung. Full raster scans across the entire tissue image were made on left lung autoradiographs from two animals killed at each time point to assess the homogeneity of fission fragment track distribution throughout the entire section. There was no evidence of any temporal change in the proportion of tracks associated with the lung periphery. At all time points PI, the track distribution was significantly nonhomogeneous, suggesting a nonuniform pattern of tissue irradiation from the 234U alpha particles. At time points from 180 to 720 days PI, large clusters of macrophages were observed in some of the sections taken from all five lung lobes. The total number of macrophage clusters increased with time PI. These macrophage clusters produced many 235U fission fragment tracks within the CR-39 autoradiographs.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of bronchopulmonary lavage on lung retention and clearance of particulate material in hamsters. Hamsters were exposed to an aerosol of fused aluminosilicate particles (FAP) labeled with 57Co. Three groups of animals were given bronchopulmonary lavage, beginning at either 1 week, 1 month, or 6 months after exposure. Each treated group was lavaged eight times over a period of 25 days. Each lavage involved 10 saline washes of the lungs. For each group, about 60-70% of the body content of 57Co at the start of lavage treatment was removed; nearly half of this was recovered in the first two lavages. A positive correlation was demonstrated between the macrophage content and 57Co activity of the washings. The subsequent fractional clearance rate of 57Co from lavaged animals was not significantly different from that in a group of untreated control animals.


Alveolar macrophages and lung lesions after combined exposure to nickel, cobalt, and trivalent chromium. In earlier inhalation exposures of rabbits, nickel increased the production of surfactant by type II cells, with secondary effects on morphology and function of alveolar macrophages. Cobalt induced mainly a nodular growth pattern of the type II cells. Trivalent chromium seemed to impair the capacity of macrophages to catabolize surfactant but did not affect the type II cells. We exposed rabbits by inhalation to combinations of nickel (0.6 mg/m3 as NiCl2) and trivalent chromium [1.2 mg/m3 as Cr(NO3)3] (Ni-Cr), cobalt (0.5 mg/m3 as CoCl2) and nickel (0.5 mg/m3) (Co-Ni), or cobalt (0.5 mg/m3) and chromium (1.2 mg/m3) (Co-Cr) for 4 months, 5 days/week, 6 hr/day. Alveolar macrophages, alveolar type II cells, and lung content of phospholipids were determined. All combined exposures induced more pronounced lung lesions than exposures for each of the metals. Phospholipid concentrations were significantly higher. There were significantly higher percentages of macrophages filled with surfactant-like inclusions and a smooth surface. Accumulations of macrophages in alveoli were more widespread. Chromium potentiated the effects of nickel and cobalt on the type II cells, which led to secondary effects on the macrophages. Nickel potentiated the specific effects of cobalt, i.e., type II cell nodule formation. The result indicates that noxious effects could also be induced in man by combined exposure to nickel, cobalt, and trivalent chromium in concentrations similar to those occurring in some occupational settings.


Epithelial and extracellular matrix injury in quartz-inflamed lung: role of the alveolar macrophage. The bronchoalveolar leukocytes from quartz-inflamed lung were separated into macrophage-enriched and neutrophil-enriched populations on density gradients. Neutrophil-enriched populations showed the greatest activity in causing injury to epithelial cells and fibronectin in vitro. Inflammatory macrophage-enriched populations from quartz-exposed lung had the ability to cause fibronectin degradation but could not cause detachment injury to epithelial cells over and above that caused by control alveolar macrophages. Fibronectin damage in vivo could be an important factor in disordering the connective tissue scaffold of the lung, thereby favoring fibrosis. In vitro quartz stimulated more production of cytokines by alveolar macrophages than the inert particulate titanium dioxide. Cytokines could be important in upregulating adhesion molecules in the membranes of lung cells in vivo; this process could aid leukocyte/lung cell contact, allowing epithelial injury to be expressed, and could also be a factor leading to pathological change.


Sequential respiratory, psychologic, and immunologic studies in relation to methyl isocyanate exposure over two years with model development. Of 113 methyl isocyanate (MIC)-exposed subjects studied initially at Bhopal, India, 79, 56, 68, and 87 were followed with clinical, lung function, radiographic, and immunologic tests at 3, 6, 12, 18, and 24 months. Though our cohort consisted of subjects at all ages showing a varied severity of initial illness, fewer females and young subjects were seen. Initially all had eye problems, but dominant symptoms were exertional dyspnea, cough, chest pain, sputum, and muscle weakness. A large number showed persistent depression mixed with anxiety, with disturbances of personality parameters. The early radiographic changes were lung edema, overinflation, enlarged heart, pleural scars, and consolidation. The persistent changes seen were interstitial deposits. Lung functions showed mainly restrictive changes with small airway obstruction; there was impairment of oxygen exchange. Oxygen exchange improved at 3-6 months, and spirometry improved at 12 months, only to decline later. The expiratory flow rates pertaining to large and medium airway function improved, but those for small airways remained low. There were changes of alveolitis in bronchoalveolar lavage fluid on fiber optic bronchoscopy, and in 11 cases positive MIC-specific antibodies to IgM, IgG, and IgE were demonstrated. On follow up, only 48% of the subjects were clinically stable, while 50% showed fluctuations. Thirty-two percent of the subjects had lung function fluctuations. Detailed sequential behavior over 2-4 years was predicted for dyspnea, forced vital capacity, maximum expiratory flow rate (0.25-0.75), peak expiratory flow rate, VO2, and depression score. A model for clinical behavior explained a total variance of 52.4% by using the factors of cough, PCO2 and X-ray zones in addition to above five parameters. The behavior of the railway colony group (1640 patients) revealed a similar pattern of illness. When this observed pattern of changes was transferred to the affected Bhopal city sections (with an equitable age-sex distribution), our model results were again validated. Thus the picture of MIC-induced disease seems similar despite the differences for age-sex and initial severity of illness in our cohort and in the population of Bhopal city as predicted by our model.


Nonrandom distribution of breakpoints in the karyotypes of workers occupationally exposed to benzene. The distribution of the breakpoints in the karyotypes of workers occupationally exposed to benzene was tested. Fifty-six workers exposed to benzene were investigated cytogenetically. A significant increase in chromosome aberrations was observed. The distribution of the breakpoints in the karyotypes of examined workers was significantly nonrandom (p less than or equal to 0.001). The breakpoints accumulated mainly on chromosomes 2, 4, and 7.


Agricultural chemical utilization and human health. The public is justifiably concerned about the human health effects of agricultural chemicals. The many gaps in information about the mechanisms of toxic action, human exposures, and the nature and extent of human health effects are large. Very few older pesticides, in particular, have been tested for human health effects. Workers who produce, harvest, store, transport, process, and prepare food and fibers are exposed to many chemicals that are potentially hazardous and that are used in agriculture. The occupational health of these workers has not been adequately studied, and protective efforts have sometimes been minimal. Valid and accurate risk assessment is best based on sound information about how chemicals, in this case agricultural chemicals, are involved in toxic events--their mechanisms of action. These health effects include tumor promotion, chronic and acute neurotoxicity, immunotoxicity, and reproductive and developmental toxicity. Another key part of risk assessment is exposure assessment. Fundamental studies of the toxicology of target organisms and nontarget organisms exposed to agricultural chemicals are needed to discover and develop better solutions to the problems of agricultural pest control, including better formulations, optimal application rates and public education in safety and alternative agricultural practices. The large number of pesticides that have never been adequately tested for effects on human health is particularly worrisome in light of emerging information about delayed nervous system effects.


Cancer risks from arsenic in drinking water. Ingestion of arsenic, both from water supplies and medicinal preparations, is known to cause skin cancer. The evidence assessed here indicates that arsenic can also cause liver, lung, kidney, and bladder cancer and that the population cancer risks due to arsenic in U.S. water supplies may be comparable to those from environmental tobacco smoke and radon in homes. Large population studies in an area of Taiwan with high arsenic levels in well water (170-800 micrograms/L) were used to establish dose-response relationships between cancer risks and the concentration of inorganic arsenic naturally present in water supplies. It was estimated that at the current EPA standard of 50 micrograms/L, the lifetime risk of dying from cancer of the liver, lung, kidney, or bladder from drinking 1 L/day of water could be as high as 13 per 1000 persons. It has been estimated that more than 350,000 people in the United States may be supplied with water containing more than 50 micrograms/L arsenic, and more than 2.5 million people may be supplied with water with levels above 25 micrograms/L. For average arsenic levels and water consumption patterns in the United States, the risk estimate was around 1/1000. Although further research is needed to validate these findings, measures to reduce arsenic levels in water supplies should be considered.


Mechanisms, measurement, and significance of lung macrophage function. Macrophages exist throughout the body. They have critical roles in the peritoneal cavity, bone marrow, skin, spleen, liver, and elsewhere. Their migratory patterns, phagocytic behavior, immunologic roles, and secretory potential are pivotal to both defense mechanisms and to the pathogenesis of disease. Macrophages have been implicated recently in such diverse disease processes as arthritis, AIDS, and juvenile onset diabetes. It is important to recognize the existence of other lung macrophages besides alveolar macrophages. Macrophages exist in small and large airways above and below the mucus. They may release chemotactic factors and a variety of mediators. They ingest and degrade antigens and are microbicidal. Interstitial macrophages are in direct contact with the extracellular matrix as well as other cells in pulmonary connective tissue such as fibroblasts. Thus, release of mediators or enzymes by interstitial macrophages can have a profound effect. Pulmonary intravascular macrophages are resident cells within the pulmonary capillaries of some species. They avidly remove particles and pathogens from circulating blood and secrete inflammatory mediators. Finally, pleural macrophages are involved in the fate and consequences of inhaled particles, especially fibers. A key attribute of macrophages is motility. Movement is an essential step in phagocytosis. There can be no particle binding or ingestion unless macrophage-particle contact occurs. To what extent and by what mechanisms do alveolar macrophages move on the alveolar epithelium? We have used optical methods as well as magnetometry to describe macrophage motility. Lung macrophages express an array of contractile proteins that are responsible for spreading, migration, phagocytosis, and the controlled intracellular motions of phagosomes and lysosomes.(ABSTRACT TRUNCATED AT 250 WORDS)


Heterogeneity of alveolar macrophages in experimental silicosis. The alveolar macrophage (AM) population has been shown to be heterogeneous in composition as well as in function. The aim of our study was to assess morphological and functional features of AM in an experimental model of quartz-induced lung fibrosis by flow cytometric methods. Twelve cynomolgus monkeys were exposed 8 hr/day, 5 days/week for 26 months to either normal atmosphere (n = 5) or 5 mg/m3 DQ12 less than 5 microns quartz dust (n = 7). After 20 months of exposure, we studied AM phagocytosis by incubating bronchoalveolar lavage cells with fluorescent polystyrene microspheres (mean diameter 1.91 microns). Using a fluorescence-activated cell sorter analyzer, AM subpopulations were identified via their volume/side scatter properties. After selective electronic "gating" of the AM populations, both the percentage of phagocytic AM and the mean number of ingested microspheres per AM were determined. In addition, a phagocytic index (microspheres/AM x % phagocytic AM x 10(-2) and a hypothetical total phagocytic capacity of one lung (phagocytic index x total number of AM x 10(-6) were calculated. The total bronchoalveolar lavage cell counts rose (75.6 +/- 11.3 x 10(6) versus 10.1 +/- 0.8 x 10(6)) significantly after quartz exposure. In contrast, the percentage of phagocytic AM was significantly (p less than 0.05) reduced (43.5 +/- 5.0% versus 74.2 +/- 1.4%). Flow cytometric measurements revealed the appearance of an AM subpopulation characterized by size/granularity features identical to blood monocytes. Only minimal numbers of these cells were found under normal conditions, but they constituted 50% of the entire AM population in the quartz group.(ABSTRACT TRUNCATED AT 250 WORDS)


Alveolar macrophage kinetics after inhalation of 239PuO2 by CBA/Ca mice: changes in synthesis of DNA. For workers in the nuclear industry, the primary route for the entry of radioactive materials into the body is by inhalation, and the rate of clearance of particles from the pulmonary region of the lung is an important factor in determining radiation dose. It is the function of alveolar macrophages (AM) to maintain the sterility of the lung and to remove insoluble particles from the respiratory surfaces and airways. The AM population is not static, and under normal conditions the loss of macrophages from the alveoli via the conducting airways is balanced by renewal. Studies of the effects of external irradiation on the kinetics of AM are numerous, but to date little is known about the effects of inhaled radioactive particles. In this investigation the effects of inhaled 239PuO2 (plutonium dioxide) particles on the synthesis of DNA by AM were studied at times up to 77 days after exposure. We also measured the number of cells recovered by bronchoalveolar lavage and the incidence of AM with nuclear aberrations. The latter provides a sensitive indicator of the effects of radiation. One of the earliest effects observed after exposure to 239PuO2 is a reduction in the number of AM recovered by lavage. This reduction is associated with a 3-fold reduction in the proportion of AM undergoing DNA synthesis at early times after exposure. The overall mean pulse labeling index of AM recovered from sham-exposed mice is 1.68%, and no trend is observed with time.(ABSTRACT TRUNCATED AT 250 WORDS)


Secretory and accessory cell functions of the alveolar macrophage. We have attempted to address the requirements necessary for alveolar macrophage accessory cell function. We have also examined the in vitro and in vivo factors that must be taken into account when interpreting results from experimental studies. Differences in phenotypic expression by rat alveolar pleural and peritoneal macrophages are noted, as well as the differing expression of major histocompatibility complex (MHC) class II molecules. Furthermore, alveolar macrophages, harvested from rat lung, do not express the interleukin (IL)-1 cytokines, and lipopolysaccharide (LPS) treatment of quiescent cells (after 24-hr in vitro culture) induces low levels of expression of IL-1 alpha and IL-1 beta. Short-term inhalation of refractory ceramic fibers, however, results in markedly increased IL-1 beta expression after stimulation with LPS. We suggest that, in vivo, IL-1 beta may be involved in the initial recruitment and activation of inflammatory cells rather than in induction of immune responses. We also postulate, based on recent published evidence, that alveolar macrophages activate the dendritic cells within the respiratory epithelium. Thus alveolar macrophages would release cytokines critical for the activation of dendritic cells during the afferent limb of the immune response, and they would respond to products of sensitized T-cells such as interferon-gamma and IL-4 to interact with T-helper cells in an antigen-specific MHC-restricted manner during the efferent limb of the response.


Persistent inflammation and impaired chemotaxis of alveolar macrophages on cessation of dust exposure. Rats were exposed by inhalation to coal mine dust, titanium dioxide, or quartz. The magnitude of the consequent inflammatory response was assessed by counting numbers and types of leukocytes in the bronchoalveolar lavage fluid. The magnitude of the inflammatory response reflected the toxicity of the dusts, with quartz eliciting the greatest recruitment of inflammatory leukocytes, coal mine dust less than quartz, and titanium dioxide eliciting no inflammation. To assess the persistence of the inflammation, groups of rats were maintained in room air for 30 or 60 days after cessation of dust exposure and then numbers of leukocytes were assessed. Bronchoalveolar leukocytes in rats exposed to coal mine dust were reduced after exposure, but in the quartz-exposed rats the numbers increased with time after exposure. The chemotactic responses of bronchoalveolar leukocytes from rats inhaling coal mine dust and quartz were reduced and remained so after a 30-day recovery period. Their reduced ability to chemotact did not fully prevent macrophages from leaving the bronchoalveolar region of dust-exposed rats. However, it is likely that the delayed removal of inflammatory leukocytes with the potential to injure the lung tissue may contribute to septal damage and so contribute to the pathogenesis of pneumoconiosis.


Effects of ozone exposure on lipid metabolism in human alveolar macrophages. Alveolar macrophages (AM) store arachidonic acid (AA), which is esterified in cellular phospholipids until liberated by phospholipase A2 or C after exposure to inflammatory stimuli. After release, there can be subsequent metabolism of AA into various potent, biologically active mediators including prostaglandins and platelet-activating factor (PAF). To examine the possibility that these mediators may account for some of the pathophysiologic alterations seen in the lung after ozone (O3) exposure, human AM were collected by bronchoalveolar lavage of normal subjects, plated into tissue culture dishes, and the adherent cells were incubated with [3H]AA or [3H]lysoPAF. Human AM exposed to 1.0 ppm O3 for 2 hr released 65 +/- 12% more tritium, derived from [3H]AA, than paired, air-exposed controls into media supernatants. In other studies using a similar O3 exposure protocol, there was also a significant increase in human AM prostaglandin E2 production (2.0 +/- 0.5-fold increase above air-exposure values, p less than 0.01, n = 17). In additional studies, using a similar O3 exposure protocol (1.0 ppm for 1 hr), there was also a significant increase in human AM PAF content (1.7 +/- 0.2-fold increase above air-exposure values, p less than 0.02, n = 5). These potent lipid mediators, originally derived from human AM, may play an important role in the mechanisms of O3 lung toxicity.


A new simplified dietary history method for measuring intake of energy and macronutrients. In this dietary history method, which preserves the characteristic features of the full dietary history, the principle of simplification is to group several food items into common codes. This enables us to complete both an interview and all nutrient calculations within 35-45 min. In this study we investigated whether the method could replace a 4-day food record for assessing individual intakes of energy and macronutrients. Dietary data were collected from 75 individuals (57 males and 18 females). Protein intake was validated by comparison with 24 h urinary nitrogen excretion (33 subjects), and energy intake was compared to weight maintenance energy intake from a later controlled diet study (19 subjects). When average intakes according to the two methods were compared, dietary history intakes were slightly higher than food record results (0-13%) except for alcohol. Comparison with urinary nitrogen excretion and energy intake data indicate that this is due to an overestimation by the dietary history method, rather than an underestimation by the food record. For individuals most of the correlations between the two methods were in the range 0.5-0.7, but alcohol correlation was down to 0.2. About 75% (53-93%) of the individuals at the extremes of the intake distribution were classified similarly by both methods. Although the ability of the dietary history to predict the individual food record results was unreliable the two methods agreed better when classifying individuals relative to certain cut-off points (sensitivity and specificity values of 0.7-0.9, relative to the food records).(ABSTRACT TRUNCATED AT 250 WORDS)


Metabolic responses to starch in bread containing intact kernels versus milled flour. In the present study, the potential of including intact kernels from different cereals was evaluated as a means of developing bread with 'lente' characteristics. Postprandial glucose and insulin responses to bread products were studied in healthy subjects. In parallel, the in-vitro enzymic starch availability was investigated. Also studied were the contents of in-vitro indigestible starch. Coarse bread (CB) products composed of 80% pre-boiled kernels from wheat, rye, oats or barley and 20% white wheat flour were baked. In the case of barley, two forms for pre-treatments was used, boiling and scalding. A bread with 80% wholemeal barley flour and 20% white wheat flour (WMB) was also included and a white wheat bread (WWB) was used as reference. The glycaemic and insulinaemic indexes (GI and II, respectively) were calculated from the 95 and 120 min incremental blood glucose and insulin areas. The GIs were significantly lower with CB from wheat, rye and barley than with WWB. In contrast, the GIs with CB from oats and WMB from barley were similar to that with WWB. The GIs and IIs were generally closely correlated. However, the II with CB from oats was significantly lower than with WWB despite similar GI. The GIs, and in particular IIs, were closely correlated with the hydrolysis rate index (HI) obtained in vitro, and this procedure can be recommended as a tool for ranking of starchy food. It is concluded that the botanical structure is an important determinant of the enzymic availability and hence of the metabolic responses. The in-vitro indigestible starch content was highest in CB from barley (1.2% dry weight basis) and lowest in CB from oats (0.5%).


Postprandial thermogenesis in obese children before and after weight reduction. The thermic effect of a meal (TEM) was measured in a group of 10 prepubertal obese children before (OB) and after (OA) weight reduction, and in a group of 10 age-matched control children (C) of normal body weight. Following a hypocaloric balanced diet for 6 +/- 1 months, the obese children lost 5.2 +/- 1.3 kg i.e. 11% of their initial body weight. The thermic response to the mixed liquid meal - fed at an energy level corresponding to 30% of the 24 h premeal resting metabolic rate - was found to be significantly lower in OB than in C children (61 +/- 25 kJ.3h-1 vs 79 +/- 21 kJ.3h-1, P less than 0.05), despite their higher test meal energy. After slimming, the TEM of obese children increased towards the controls' values (73 +/- 30 kJ.3h-1). These results support the hypothesis of the existence of a moderate thermogenic defect in some obese children which represents a consequence rather than an aetiological factor of obesity.


The variation in blood lipid levels described by various measures of overall and abdominal obesity in Danish men and women aged 35-65 years. The purpose of the study was to describe the proportion of the variation in blood lipid levels [high-density lipoprotein cholesterol (HDLC), low-density lipoprotein cholesterol (LDLC), total cholesterol, very-low-density lipoprotein cholesterol (VLDLC) and triglycerides] explained by different measures of overall obesity [body fat (kg), percentage body fat, or body mass index (kg/m2)] and abdominal obesity [waist/hip ratio, waist/thigh ratio or waist circumference (cm)]. This was done in a Danish population sample of 1523 men and 1464 women aged 35-65 years. This was done to assess, on a population level, the effects on the different lipid levels to be expected from a possible reduction in the level of obesity. The proportion of the variation in lipid levels explained by the various measures of overall obesity differed only slightly, as did the proportion of the variation in lipid levels explained by the various measures of abdominal obesity. In men more of the variation in the blood lipids could be explained by overall obesity than by abdominal obesity, whereas in women the reverse was true. More of the variation in the lipids was explained by overall obesity in men than in women, but more of the variation was explained by abdominal obesity in women than in men. In women the obesity measures predicted between 0% and 11% of the variation in lipid level, and in men the obesity measures predicted between 0% and 14% of the variation. Between 16% and 30% in women and between 5% and 21% in men of the variation in the lipid levels could be explained by obesity, age and several lifestyle variables.(ABSTRACT TRUNCATED AT 250 WORDS)


Obesity: an inherited metabolic deficiency in the control of macronutrient balance? It has generally been assumed that the body is 'energy blind' and calories from all three macronutrients contribute with the same value to energy balance. There is, however, accumulating evidence to suggest that during ad-libitum conditions energy balance is achieved by a separate regulation of carbohydrate, fat and protein balances. Regulation of carbohydrate balance has the highest priority in the hierarchy, which is appropriate because the limited glycogen stores are only capable of covering the carbohydrate oxidation for a few days. Due to the higher satiating power of carbohydrate and protein compared with fat, a reduction in the dietary fat/carbohydrate ratio produces a negative fat balance in normal subjects consuming the diet ad libitum, while an increase in dietary fat/carbohydrate ratio results in a positive fat balance and weight gain. Subjects with a genetically determined predisposition to obesity become obese when they are exposed to a particular range of environmental conditions. The available knowledge suggests that the genetic propensity to weight gain is caused by a susceptibility to dietary fat due to an impaired capacity to increase their lipid/carbohydrate oxidation when fed a high-fat/low-carbohydrate diet. This in turn promotes lipid storage, depletion of carbohydrate stores and increases appetite. By enlarging the fat stores, the accompanying insulin resistance and higher levels of circulating non-esterified fatty acids increase lipid oxidation until it is commensurate with the dietary fat intake. The development of obesity may therefore be viewed as a regulatory mechanism by which the impaired lipid oxidation rate is raised to match a high fat intake. However, by decreasing the dietary ratio of fat to carbohydrate, macronutrient balance may be achieved with a high energy expenditure and a normal body composition. The results support current dietary recommendations, but with less emphasis on carbohydrate source, and they are also applicable for the prevention and treatment of obesity.


Effects of an oat bran concentrate on serum lipids in free-living men with mild to moderate hypercholesterolaemia. An oat bran concentrate was prepared by removing non-fibre components by cold-water wet-milling, resulting in a 2- to 3-fold concentration of soluble fibre, with beta-D-glucan as its main component. The concentrate was baked in bread which was consumed for 8 weeks by free-living men with mild to moderate hypercholesterolaemia. The effects on serum lipids were assessed in a randomized, double-blind, placebo-controlled trial. Despite the large daily dose (11.2 g) of beta-glucan, the beta-glucan-enriched bread had only a small and statistically non-significant effect on serum lipid concentrations. Probable reasons for the weakness of the effect could be the poor solubility of beta-glucan in the preparation, its enzymatic hydrolysis after ingestion, and the consequently low viscosity in the intestine.


Influence of sterilization, drying and oat bran enrichment of pasta on glucose and insulin responses in healthy subjects and on the rate and extent of in vitro starch digestion. The effects of sterilization and oat bran enrichment of pasta on the glucose and insulin responses in healthy subjects were evaluated. Cooked and canned spaghetti and cooked fettucini without and with enrichment with oat bran (28%) were compared. Further, the effects of various low- and high-temperature drying conditions for spaghetti, cooking time and sterilization on the starch digestion rate and content of enzyme-resistant starch (RS) in vitro were also studied. Various cooking quality data were also determined to allow interpretation of results. The incremental glucose area (0-120 min) produced by canned spaghetti was twice the area of that produced by cooked spaghetti (69.03 vs 35.45 mmol/l x min, P less than 0.01). The incremental insulin area (0-120 min) was also significantly higher with canned spaghetti (17,500 vs 12,600 pmol/l x min, P less than 0.05). The rapid digestion was caused by excessive swelling of starch during sterilization that promoted a very soft texture of the spaghetti. Enrichment of fettucini with oat bran reduced slightly the incremental insulin area (15,600 vs 20,100 pmol/l x min, P less than 0.05, for 0-120 min), but did not significantly reduce the glucose area. Drying conditions and cooking times could be varied within broad limits without affecting the rate of starch digestion in vitro of cooked spaghetti. In sterilized spaghetti the content of resistant starch was higher than that found in cooked 'al dente' spaghetti (2.2-3.4 vs 0.5 mg/100 mg total starch). In conclusion, sterilization influences the nutritional properties of starch in pasta by substantially increasing the glucose and insulin responses and by formation of resistant starch. The effect of oatbran environment is restricted mainly to a slight decrease in the insulin response.


An in vitro procedure based on chewing to predict metabolic response to starch in cereal and legume products. A new method for measuring the rate of in-vitro starch digestion in products with a structure 'as eaten' is introduced. An equivalent amount of potentially available starch from each product was chewed by subjects, expectorated into a beaker and incubated with pepsin. The incubate was thereafter transferred to a dialysis tubing and incubated with pancreatic alpha-amylase for 3 h. Samples were removed from the dialysate at time intervals and the degree of hydrolysis was calculated as the proportion of the potentially available starch degraded to maltose. A hydrolysis index (HI) was calculated as the area under the hydrolysis curve with the product as a percentage of the corresponding area with white wheat bread. The method was applied to 21 cereal and legume products, chosen to cover as wide a range as possible with respect to metabolic response, and to include several of the proposed mechanisms to differences in metabolic behaviour of starch. The accuracy of the in-vitro method was evaluated versus the metabolic responses obtained with the same products in healthy subjects. A significant correlation between HI and glycaemic index (GI) was obtained in cereal as well as in legume products. A significant correlation was also obtained between HI and insulin index (II) with pooled data from all products. However, in the case of II no correlation was obtained with the legume products only. It is concluded that the presently described in-vitro procedure offers a good potential to predict the metabolic behaviour of starchy foods.


The effects of a 2-year switch from a mixed to a lactovegetarian diet on trace element status in hypertensive subjects. Trace element status of 20 hypertensive subjects (14 women and 6 men, mean age 52 years) was compared to that of normotensives. The changes in trace element status, body weight and blood pressure after a 2-year switch from a mixed to a lactovegetarian diet were also compared between these groups. The concentration of copper in plasma and that of lead in hair were higher in hypertensive subjects than in normotensives, but the concentrations of zinc, magnesium and selenium in plasma, urine and hair were similar to those of normotensives. In the hypertensive subjects, 3 months after the diet shift there was a decrease in the concentrations of zinc in plasma, hair and urine, that of copper in plasma and hair, of magnesium in urine, of selenium in plasma and hair, and an increase in the magnesium content in plasma and hair. Also the concentrations of mercury, lead and cadmium in hair decreased after the diet switch. Among the hypertensives, the relative increase of magnesium in plasma was greater than that of normotensives; their relative decreases of selenium and lead in hair were lower, that of cadmium greater and that of copper in urine was lower. Four years after the start of the experiment when most subjects had resumed a mixed diet, mineral and trace element concentrations in plasma, hair and urine were similar to their baseline levels. Three months after the diet switch the relative decrease in body weight was more marked among hypertensive women (5%) than in female normotensives (3%), but similar among men of both groups (5%).(ABSTRACT TRUNCATED AT 250 WORDS)