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E Esposito

Publications and source records attributed to E Esposito.

At least 37 records · Page 2Linked to original sources

Pectin-based microspheres: a preformulatory study.

This paper reports on (a) the production of pectin microspheres and (b) the influence of different experimental parameters and ionic crosslinking on morphological and dimensional characteristics of pectin microspheres. Morphological and dimensional characteristics of pectin were analyzed as a function of the type of pectin, the dispersing phase, the stirring speed, and the emulsifying agent. Crosslinking by calcium chloride and the encapsulation of antibiotics (i.e., metronidazol and tetracycline) gave particles morphologically similar to empty particles but with slower swelling kinetic.

Chemistry, Pharmaceutical↗

Liposomes as carriers for DNA-PNA hybrids.

Peptide nucleic acids (PNAs) are DNA mimics composed of N-(2-aminoethyl)glycine units. This structure gives to PNAs (a) resistance to DNases and proteinases, (b) capacity to hybridize with high affinity to complementary sequences of single-stranded RNA and DNA, and (c) capacity to form highly stable (PNA)(2)-RNA triplexes with RNA targets. Furthermore, DNA-PNA hybrid molecules are capable to reversibly interact with DNA-binding proteins, being therefore of interest for studies on regulation of gene expression by the decoy approach. The major conclusion of this paper is that cationic liposomes are able to efficiently complexate DNA-PNA hybrid molecules and mediate their binding to target cells. Our results are of some interest, since, unlike commonly used nucleic acids analogs, PNA oligomers are not taken up spontaneously into the cells. In addition, they are not suitable for an efficient delivery with commonly used liposomal formulations. Transfection of PNA-DNA hybrid molecules to in vitro cultured cells could be of great interest to determine the applications of these new reagents to experimental alteration of gene expression.

DNA↗

Biochemical and electrophysiological evidence that RO 60-0175 inhibits mesolimbic dopaminergic function through serotonin(2C) receptors.

In vivo microdialysis and electrophysiological techniques were used to elucidate the role of the 5-HT(2) receptor family on the control of mesolimbic dopaminergic system exerted by serotonin (5-HT). Administration of RO 60-0175 (1 mg/kg, i.p.), a selective 5-HT(2C) receptor agonist, significantly decreased dopamine (DA) release by 26+/-4% (below baseline) 60 min after injection. Moreover, RO 60-0175 (80-320 microg/kg, i.v.) dose-dependently decreased the basal firing rate of DA neurons in the ventral tegmental area (VTA), reaching its maximal inhibitory effect (53.9+/-15%, below baseline) after the dose of 320 microg/kg. The selective 5-HT(2C) receptor antagonist SB 242084 completely blocked the inhibitory action of RO 60-0175 on accumbal DA release and on the firing rate of VTA DA cells. On the contrary, both (+/-)-DOI, a mixed 5-HT(2A/2C) receptor agonist, and the selective 5-HT(2B) agonist BW 723C86, did not affect either DA release in the nucleus accumbens or the firing rate of VTA DA cells. Taken together, these data confirm that central 5-HT system exerts an inhibitory control on the mesolimbic DA system and that 5-HT(2C) receptors are involved in this effect.

Animals↗

Influence of formulation and process parameters on pellet production by powder layering technique.

The goal of the present study was to evaluate the influence of the formulation and operating conditions on pellet preparation by pan technique. To this end, a new pelletization process, typified by the application of powdered drug on sugar-based cores using the GS coating system was studied. Inert cores were intermittently treated with micronized drug powder and adhesive solution. This treatment led to the formation of multiple layers of drug particles around an inert core resulting in the production of pellets that can further be coated by different polymers to obtain modified release formulations. Different procedures have been used to evaluate a series of important parameters such as initial core weight; speed of powder application; speed, type, and position of the atomizers; atomization degree; temperature; and air cap. Good yield of drug layering was obtained by adjusting the quantity of both the drug powder to apply and the binder solution. Pellets obtained following the optimal operating conditions (defined in a pre-formulation study) were film coated with the acrylic polymer Eudragit L30D in order to produce a model formulation consisting of enteric polymer-coated pellets containing ibuprofen. During its preparation, the formulation showed no degradation of the drug; moreover, a low percentage of residual humidity was obtained, indicating that this system is very efficient for the production of highly stable formulations. This study showed the good performance of the GS automated pan-coating system in obtaining enteric coated pellets prepared by powder layering technique using aqueous solutions.

Chemistry, Pharmaceutical↗

Preferential modulation of mesolimbic vs. nigrostriatal dopaminergic function by serotonin(2C/2B) receptor agonists: a combined in vivo electrophysiological and microdialysis study.

Electrophysiological and in vivo microdialysis were used to investigate and compare the effect of tonic activation of serotonin(2C/2B) (5-HT(2C/2B)) receptors on nigrostriatal and mesolimbic dopaminergic (DA) function. Thus, extracellular single unit recordings of neurochemically-identified DA neurons in the SNc and the VTA, as well as simultaneous monitoring of striatal and accumbal DA release were performed following the administration of the unselective 5-HT(2C/2B) agonists, mCPP (m-chlorophenylpiperazine) and MK 212 [6-chloro-2-(1-piperazinyl)piperazine]. Both mCPP (5-320 microg/kg i. v.) and MK 212 (5-320 microg/kg i.v.) dose-dependently decreased the firing rate of VTA DA neurons. The maximal effect was reached at the cumulative dose of 320 microg/kg mCPP and MK 212, which caused a decrease of 42.6 +/- 12.8% and 56.4 +/- 12.6%, respectively. In addition, the total number of events in bursts and the number of bursts of VTA DA cells were significantly reduced by both mCPP and MK 212. On the other hand, mCPP (5-320 microg/kg i.v.) and MK 212 (5-320 microg/kg i.v.) induced a slight decrease in the basal firing rate, but not in bursting activity of SNc DA neurons. Consistent with electrophysiological data, dialysate DA levels in the nucleus accumbens decreased significantly, reaching the maximum of 26.6 +/- 9.6% below baseline levels 120 min after mCPP (1 mg/kg i.p.) administration, and of 25.2 +/- 5.5% 140 min after MK 212 (1 mg/kg i. p.) injection. DA outflow in the striatum was unaffected by both drugs. The inhibitory effect of both mCPP and MK 212 on VTA DA cell activity was blocked completely by pretreatment with the selective 5-HT(2C) antagonist SB 242084 ¿6-chloro-5-methyl-1-[2-(2-methylpyridyl-3-oxy)-pyrid-5-yl carbamoyl] indoline¿ (200 microg/kg), given intravenously 10 min before the first injection of the 5-HT(2C/2B) agonists. SB 242084 (2. 5 mg/kg i.p.) antagonized also the decrease in DA release induced by mCPP and MK 212 in the nucleus accumbens. Taken together, these data indicate that mCPP and MK 212 selectively inhibit mesolimbic dopaminergic function by acting on 5-HT(2C) receptors. Therefore, selective 5-HT(2C) receptor agonists might be useful in clinical conditions where it is necessary to reduce the mesolimbic dopaminergic activity without affecting the nigrostriatal function.

Aminopyridines↗

Rheologic and NMR characterization of monoglyceride-based formulations.

This paper describes the production and characterization of semi-solid formulations based on monoglycerides from canola oil and water as drug-delivery systems. In order to obtain new formulations with different characteristics in terms of viscosity, bioadhesiveness, and solubilization capacity, a third component was added to the monoglyceride-water system. Nine excipients were tested, namely soy oil, isopropylmyristate, isopropylpalmitate, tripalmitin, tristearin, glyceryl monostearate, glycerol, propylene glycol, and ethanol. In particular, the effect of each excipient on the viscosity and stability of the formulation was investigated. It was found that ethanol dramatically influenced the viscosity of the monoglyceride-water system, resulting in the formation of stable forms. In addition, ethanol suitably could be used for the solubilization of water-insoluble lipophilic drugs. This promising ternary system was characterized by microscopic, NMR spectroscopic, and rheologic techniques. (1)H and (13)C NMR studies were made of Myverol to verify the molecular structure and isomer distributions of this commercial monoacylglycerol mixture. The microstructure of an isotropic solution consisting of Myverol [1.8% (w/w)], ethanol (42.9%), and water (55.3%) was studied by the multi-component self-diffusion NMR method. From the self-diffusion coefficient (D) of the monoglycerides (8.8 x 10-11 m(2)/s), an "apparent spherical hydrodynamic radius" of ca. 2.48 nm was calculated for the micellar aggregate. A nearly spherical shape is consistent with these values since the extended hydrocarbon chain of the longest monoglyceride (17 carbons) is ca. 2.2 nm long. The D's of water and ethanol reveal they do not associate (no attractive nonbonding interactions) appreciably with the fatty acid micelles.

Biocompatible Materials↗

Acute administration of amitriptyline and mianserin increases dopamine release in the rat nucleus accumbens: possible involvement of serotonin2C receptors.

Previous studies of conventional tricyclic and non-tricyclic antidepressants have suggested that a number of these drugs display considerable pharmacological activity at 5-HT2C receptors in the brain. There is evidence that 5-HT2C receptors are involved in the control of the activity of the central dopaminergic system. Therefore, the effects of amitriptyline (5 mg/kg and 10 mg/kg i.p.) and of the atypical antidepressant mianserin (2.5 mg/kg and 5 mg/kg i.p.) were studied on the extracellular concentration of dopamine (DA) in the nucleus accumbens of chloral hydrate-anesthetized rats, using intracerebral microdialysis. Amitriptyline and mianserin significantly increased DA release (+31.1 +/- 7.9% and +33.6 +/- 4.3%, respectively) at the higher doses. In addition, lower doses of mianserin (2.5 mg/kg i.p.) and amitriptyline (5 mg/kg i.p.) blocked the inhibitory action of RO 60-0175 (1 mg/kg i.p.), a selective 5-HT2C receptor agonist, on DA release. The effect of RO 60-0175 (1 mg/kg i.p.) was completely blocked by SB 242084 (2.5 mg/kg i.p.), a selective and powerful 5-HT2C receptor antagonist. Taken together, these data indicate that amitriptyline and mianserin increase DA release in the nucleus accumbens by blocking 5-HT2C receptors.

Adrenergic Uptake Inhibitors↗

In vitro antiproliferative activity of isothiocyanates and nitriles generated by myrosinase-mediated hydrolysis of glucosinolates from seeds of cruciferous vegetables.

A comparison of the effect of isothiocyanates and nitriles derived from some glucosinolates, namely, epi-progoitrin, sinalbin, glucotropaeolin, glucocheirolin, and glucoraphenin, on human erythroleukemic in vitro cultured cells was studied. Many studies have in fact evidenced that a consumption of vegetable containing glucosinolates could reduce the development of colorectal cancer. In the experimental conditions used, the production of isothiocyanates and nitriles from glucosinolates is almost quantitative as confirmed by HPLC or GC-MS analysis. The obtained results demonstrated that in general nitriles are considerably less potent than the corresponding isothiocyanates in inhibiting cancer cell growth. Particularly, the isothiocyanates inhibitory activity on K562 cells growth is higher in the case of products derived from epi-progoitrin, glucotropaeolin, glucoraphenin, and glucocheirolin; while for nitriles the higher activity in inhibiting K562 cells growth is showed by sinalbin-derived product. Considering the antiproliferative activity found for isothiocyanates and nitriles, further studies will be aimed to the possible application of glucosinolate-derived products as chemopreventive cancer agents for the reduction of colorectal cancer.

Brassicaceae↗

Effect of acute administration of hypericum perforatum-CO2 extract on dopamine and serotonin release in the rat central nervous system.

The hydromethanolic extract of Hypericum perforatum has been shown to be an effective antidepressant, although its mechanism of action is still unclear. In this study, in vivo microdialysis was used to investigate the effects of Hypericum perforatum-CO2 extract on dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) release in various areas of brain. Administration of Hypericum perforatum extract (1 mg/kg, p.o.) caused a slight, but significant increase of DA outflow both in the nucleus accumbens and the striatum. The maximal increase of DA efflux (+19.22+/-1.93%, relative to the control group) in the nucleus accumbens occurred 100 min after administration of Hypericum perforatum. In the striatum, the extract maximally enhanced DA outflow (+24.83+/-7.49 %, relative to the control group) 80 min after administration. Extraneuronal DOPAC levels were not significantly affected by Hypericum perforatum treatment. Moreover, Hypericum perforatum (1 mg/kg, p.o.) did not produce any significant effect on either 5-HT or 5-HIAA efflux in the ventral hippocampus. This study shows for the first time that Hypericum perforatum extract is capable of increasing in vivo DA release.

3,4-Dihydroxyphenylacetic Acid↗

Production of Eudragit microparticles by spray-drying technique: influence of experimental parameters on morphological and dimensional characteristics.

The aim of this study was to evaluate the influence of operating parameters on the characteristics of methacrylate microparticles prepared by spray-drying technique. Eudragit microparticles were prepared by a spray-drying method. The influence of different experimental parameters (i.e., solvent, feed rate, air flow rate, air-drying temperature, and aspiration flow rate) on microparticle morphology, size distribution, and recovery was studied. In addition, different Eudragit types and Eudragit RS concentrations were employed. Optical and electron microscopy were employed to analyze microparticle morphology and dimensional distribution. Finally, prednisolone as model drug was encapsulated in Eudragit RS microparticles. Low feed rate values yielded the best results in terms of microparticle morphology. Changes in nebulizing air flow did not result in a corresponding effect on microparticle characteristics. An increase of air-drying temperature resulted in a reduction of microparticle dimension and recovery. A low flow of drying air was preferable because this resulted in microparticles with an optimal morphology. The polymer concentration affected both morphology and dimensions of microparticles. The encapsulation of prednisolone led to good incorporation efficiencies without altering percentage of recovery, morphology, and mean dimension of the microparticles. The selection of appropriate parameters yielded spray-dried Eudragit RS microparticles characterized by good morphology and narrow dimensional distribution.

Acrylates↗

Effect of DNA complexation and freeze-drying on the physicochemical characteristics of cationic liposomes.

We describe the use of saccharides, such as sorbitol, mannitol, sucrose, maltodextrin, and dextran, as cyoprotectants for freeze-drying cationic liposomes. Saccharides can protect liposomes either by interacting with phospholipid headgroups or by forming an amorphous glass surrounding the vesicles, thus preventing aggregation, mechanical rupture of membrane, fusion of liposomes, and drug leakage. We have particularly considered liposome characteristics, such as size, zeta potential, and ability in complexing DNA, before and after freeze-drying. Our study indicates that cationic liposomes are able to maintain liposome characteristics after lyophilization and rehydration and maintain the ability to complex DNA even if the strength of the interaction forces was of lower intensity with respect to liposomes before lyophilization.

Binding Sites↗

Urinary BTA-stat, BTA-trak and NMP22 in surveillance after TUR of recurrent superficial transitional cell carcinoma of the bladder.

OBJECTIVES: To evaluate NMP22, BTA-Stat and BTA-Trak tests in monitoring recurrent transitional cell carcinoma of the bladder. METHODS: The tests were performed in 179 selected patients being followed up for recurrent superficial bladder tumors: 55 patients had bladder recurrence and 124 patients were recurrence free. The NMP22 test was obtained in all patients; BTA-Stat and BTA-Trak in the last 96 and 74 patients, respectively. Sixty-four patients (51.6%) were undergoing adjuvant intravesical chemotherapy. RESULTS: Sensitivity was 74, 57 and 62% and specificity was 55, 62 and 79% for NMP22, BTA-Stat and BTA-Trak, respectively. A high percentage of patients submitted to intravesical chemotherapy had false-positive tests. Positive predictive values of the NMP22, BTA-Stat and BTA-Trak tests were 42.2, 40 and 45.4%, and negative predictive values were 82.9, 76.9 and 88.4%, respectively. CONCLUSIONS: NMP22, BTA-Stat and BTA-Trak tests cannot replace cystoscopy and cannot be adopted as routine tools in surveillance after TUR in patients with superficial bladder cancer.

Adult↗

[Prophylactic postoperative radiotherapy in the treatment of supraglottic tumors].

The appropriateness of treatment of supraglottic carcinoma with post-operative radiotherapy is still a controversial issue. The purpose of the present work has been to define the effectiveness of post-operative radiotherapy in the treatment of supraglottic carcinomas and to discuss, on the basis of the data reported in the literature, the usefulness of combined surgery-radiotherapy. The study involved 97 subjects suffering from spinocellular carcinoma of the laryngeal vestibule (95 males, 2 females; average age: 58; age range: 38-89 years) who underwent horizontal supraglottic laryngectomy together with bilateral laterocervical lymph node dissection. Of these patients 35 (36%, group A) had undergone a cycle of prophylactic radiotherapy (60-70 Gy in fractions of 2 Gy/die, administered with two side fields), after surgery. The remaining 62 cases (64%, group B) did not have any additional therapy after surgery. For both groups the overall actuarial survival and 5-year corrected survival rates were calculated; statistical significance was calculated using the Wicoxon test. For group A the overall actuarial survival and corrected actuarial survival rates were, respectively, 74% and 90%. For group B these rates were, respectively 61% and 80%. A statistical comparison of both parameters did not show any statistically significant difference (p = 0.2 and 0.4, respectively). In reference to tumor extension, established both on the basis of clinical (T) and surgical-anatompatholgical (pT) findings, no significant differences were encountered between the overall actuarial survival and corrected actuarial survival rates for the two groups of patients (p > 0.05). In NO patients significantly higher overall actuarial survival rate in those patients who underwent the combined surgery-radiotherapy treatment than in those treated with surgery alone (p = 0.01) was seen. This was not, however, confirmed by the corrected actuarial survival rates and comparative analysis of the groups for surgical staging of the lymph node metastases (p > 0.05). Most likely the different behavior seen in the two groups depends on possible errors in the clinical staging of N due to the presence of reactive lymphadenitis and micrometastases. In conclusion, the data derived from the present research indicate that post-operative radiotherapy, performed as prophylaxis in cases undergoing supraglottic laryngectomy does not yield any statistically significant improvement in prognosis. Therefore, the radiotherapy association should be ruled out in those patients where the oncological radicality of the surgery is reasonably certain. It can possibly be considered as integration to surgery, only in those cases where radicality is in doubt.

Adult↗

Preparation and characterization of cationic microspheres for gene delivery.

The production and characterization of cationic microparticles based on Eudragit RS and cationic agents (i.e. a cationic acrylic polymer and three different cationic surfactants) for the delivery of nucleic acids is here described. It was found that morphological and dimensional characteristics of microparticles were influenced by the type and concentration of cationic agent employed and by some experimental parameters such as stirring speed, emulsifying agent and type of rotor. The desoxiribonucleotide Defibrotide (DFT) was associated with positively charged microparticles and its in vitro release kinetics from microparticles were determined. A study of the in vitro toxicity of cationic microparticles on cultured human cell line K562 was also performed, demonstrating that DDAB(18) microparticles display very low cytotoxicity.

Acrylic Resins↗

SB 242084, a selective serotonin2C receptor antagonist, increases dopaminergic transmission in the mesolimbic system.

Electrophysiological techniques and in vivo microdialysis were used to investigate the effect of SB 242084, a potent and selective 5-HT2C receptor antagonist in the control of nigro-striatal and mesolimbic dopaminergic function. Thus, extracellular single unit recordings were performed from neurochemically-identified dopamine (DA) neurons in the substantia nigra, pars compacta (SNc) and the ventral tegmental area (VTA), as well as monitoring of striatal and accumbal basal DA release in anesthetized rats following the administration of SB 242084 and RO 60-0175. Administration of SB 242084 (160-640 microg/kg, i.v.) caused a dose-dependent increase in the basal firing rate of VTA DA neurons, reaching its maximum (27.8+/-6%, above baseline) after 640 microg/kg. Moreover, bursting activity was significantly enhanced by SB 242084 in the VTA. On the other hand, SB 242084 (160-640 microg/kg, i.v.) did not cause any significant change in the basal firing rate and bursting activity of DA neurons in the SNc. Injection of the 5-HT2C receptor agonist RO 60-0175 (80-320% microg/kg, i.v.) dose-dependently decreased the basal firing of DA neurons in the VTA but not in the SNc. RO 60-0175 exerted its maximal inhibitory effect (53.9+/-15.1%, below baseline) in the VTA at the dose of 320 microg/kg. Basal DA release (34.8+/-9%, above baseline) and dihydroxyphenylacetic acid (DOPAC) efflux (19.7+/-7%, above baseline) were significantly enhanced in the nucleus accumbens following the intraperitoneal administration of 10 mg/kg SB 242084. Intraperitoneal injection of 5 mg/kg SB 242084 significantly increased DA release (16.4+/-6%, above baseline) in the nucleus accumbens, but did not affect DOPAC efflux. In the striatum, SB 242084 (5 and 10 mg/kg, i.p.) only slightly increased DA release above baseline (3.5+/-4 and 11.2+/-6%, respectively), without affecting DOPAC efflux in this area. However, the effect of SB 242084 in the striatum was rendered more evident by the fact that injection of the vehicle used to dissolve the drug in a group of control rats, significantly reduced basal DA output by 19.6+/-7%. Stimulation of 5-HT2C receptors by RO 60-0175 (1 mg/kg, i.p.) significantly decreased DA release in the nucleus accumbens by 26.1+/-4% (below baseline) 60 min after injection. On the other hand, RO 60-0175 (1 mg/kg, i.p.) did not cause any significant change of DA release in the striatum. However, DOPAC efflux was reduced by RO 60-0175 (1 mg/kg, i.p.) both in the striatum and the nucleus accumbens. Taken together, these data indicate that the central 5-HT system exerts a tonic and phasic inhibitory control on mesolimbic DA neuron activity and that 5-HT2C receptor subtypes are involved in this effect. Moreover, these findings might open new possibilities for the employment of 5-HT2C receptor antagonists in the treatment of neuropsychiatric disorders related to a hypofunction of central DA neurons.

Aminopyridines↗

Selective blockade of serotonin-2C/2B receptors enhances mesolimbic and mesostriatal dopaminergic function: a combined in vivo electrophysiological and microdialysis study.

Electrophysiological techniques and in vivo microdialysis were used to investigate the relative contribution of central serotonin-2C/2B and serotonin-2A receptor subtypes in the control of mesolimbic and nigrostriatal dopaminergic function. Thus, extracellular single-unit recordings were performed from neurochemically identified dopamine neurons in the ventral tegmental area and the substantia nigra pars compacta, as well as simultaneous monitoring of accumbal and striatal basal dopamine release in anesthetized rats following the administration of serotonin-2C/2B (SB 206553), serotonin-2A (SR 46349B) or serotonin-2A/2B/2C (ritanserin) antagonists. Administration of SB 206553 (40-160 microg/kg, i.v.) caused a dose-dependent increase in the basal firing rate of ventral tegmental area and nigral dopamine neurons, reaching its maximum (45.2 and 28.5%, respectively) following 160 microg/kg. Moreover, burst activity was significantly enhanced by SB 206553 in the ventral tegmental area only. In contrast, injection of SR 46349B (40-160 microg/kg, i.v.), and ritanserin (40-160 microg/kg, i.v.) did not cause any significant change in the basal activity of these neurons. Basal dopamine release was significantly enhanced in both the nucleus accumbens (42%) and the striatum (33%) following the intraperitoneal administration of 5 mg/kg SB 206553. In contrast, SR 46349B (0.5 mg/kg, s.c.) and ritanserin (0.63 mg/kg, i.p.) failed to affect basal dopamine output in both regions. Taken together, these data indicate that the central serotonergic system exerts a tonic inhibitory control of mesolimbic and nigrostriatal dopaminergic pathway activity and that the serotonin-2C/2B receptor subtypes are involved in this effect. Moreover, these findings might open new possibilities for the employment of serotonin-2C/2B receptor antagonists in the treatment of neuropsychiatric disorders related to a hypofunction of central dopaminergic neurons.

Animals↗

Reduced chaos of interspike interval of midbrain dopaminergic neurons in aged rats.

In this study, the nonlinear prediction method combined with Gaussian-scaled surrogate data was used to quantify, as a first goal, the chaotic behavior of the interspike interval of ventral tegmental area dopaminergic neurons, extracellularly recorded in vivo, in anesthetized rats. The second goal was to determine the differences in chaotic content as a function of age. Comparisons were made among three different groups of rats: young (two to four weeks of age), adult (three to four months of age) and aged (16-19 months of age). It has been found that the degree of complexity of action potential trains is reduced with aging. The chaotic content of ventral tegmental area dopamine neurons within each group and the decrease of chaos with aging cannot be explained in terms of standard characteristics of neuronal activity (firing rate, bursting activity). These data can be rationalized in the light of recent findings on the role of deterministic chaos in the functional behavior of complex biological systems, and suggests that nonlinear analysis may provide an additional method in characterizing neuronal activity.

Aging↗