Using Electroencephalogram-Extracted Nonlinear Complexity and Wavelet-Extracted Power Rhythm Features during the Performance of Demanding Cognitive Tasks (Aristotle's Syllogisms) in Optimally Classifying Patients with Anorexia Nervosa.

Anorexia nervosa is associated with impaired cognitive flexibility and central coherence, i.e., the ability to provide an overview of complex information. Therefore, the aim of the present study was to evaluate EEG features elicited from patients with anorexia nervosa and healthy controls during mental tasks (valid and invalid Aristotelian syllogisms and paradoxes). Particularly, we examined the combination of the most significant syllogisms with selected features (relative power of the time-frequency domain and wavelet-estimated EEG-specific waves, Higuchi fractal dimension (HFD), and information-oriented approximate entropy (AppEn)). We found that alpha, beta, gamma, theta waves, and AppEn are the most suitable measures, which, when combined with specific syllogisms, form a powerful tool for efficiently classifying healthy subjects and patients with AN. We assessed the performance of triadic combinations of "feature-classifier-syllogism" via machine learning techniques in correctly classifying new subjects in these two groups. The following triads attain the best classifications: (a) "AppEn-invalid-ensemble BT classifier" (accuracy 83.3%), (b) "Higuchi FD-valid-linear discriminant" (accuracy 75%), (c) "alpha amplitude-valid-SVM" (accuracy 83.3%), (d) "alpha RP-paradox-ensemble BT" (accuracy 85%), (e) "beta RP-valid-ensemble" (accuracy 85%), (f) "gamma RP-valid-SVM" (accuracy 85%), and (g) "theta RP-valid-KNN" (accuracy 80%). Our findings suggest that anorexia nervosa has a specific information-processing style across reasoning tasks in the brain as measured via EEG activity. Our findings also contribute to further supporting the view that entropy-oriented, i.e., information-based features (the AppEn measure used in this study) are promising diagnostic tools (biomarkers) in clinical applications related to medical classification problems. Furthermore, the main EEG-specific frequency waves are extremely enhanced and become powerful classification tools when combined with Aristotle's syllogisms.

Early auditory-evoked potentials in body dysmorphic disorder: An ERP/sLORETA study.

Body dysmorphic disorder (BDD) is characterized by excessive preoccupation with imagined or slight physical defects in appearance. BDD is associated with cognitive impairments (attention, visual processing). Our study aims to evaluate the early neural responses (N100, P200) to prepulse inhibition (PPI) and prepulse facilitation (PPF), to investigate attentional processing of BDD in the auditory domain. Fifty-five adults took part: 30 BDD patients and 25 healthy controls. We compared their brain responses to PPI and PPF by analyzing global field power (GFP), event-related potentials (ERPs) and their respective sources. BDD exhibited reduced N100 amplitudes compared to healthy controls in response to the startle tone elicited by both PPI and PPF, potentially suggesting impaired allocation of attention. Interestingly, the lower the GFP at the N100, the higher the BDD severity. Source reconstruction analysis showed reduced activation for BDD during the N100 time window in PPI. Scalp responses and source activations in PPI were decreased overall compared to PPF, confirming the gating effect of PPI. We provided evidence that the N100 may serve as an electrophysiological marker of BDD, predicting its severity. Our study demonstrated the potential of using ERPs combined with behavioural PPI and PPF protocols to advance our understanding of BDD pathophysiology.

Test-retest reliability of brain oscillations in a prepulse inhibition and facilitation paradigm: effects of gender in healthy humans.

There is a growing interest in assessing the reliability of electroencephalographic (EEG) measures in clinical and research settings. Prepulse inhibition (PPI: representing attentional modulation) and facilitation (PPF: reflecting selective attention) paradigms have been used to study inhibitory function and selective attention, respectively. However, to date, little has been known with regards to the stability of brain oscillatory activity during PPI and PPF. We investigated the stability of event-related EEG oscillations during PPI and PPF in healthy humans over two monthly sessions. Power spectral densities were analysed at traditional frequency bands (delta, alpha, beta sub-bands, and gamma). We assessed test-retest reliability by calculating intraclass correlation coefficients (ICCs, absolute agreement definition) and examined potential effects of gender. The results showed good-to-excellent reproducibility of EEG power (both in PPI and PPF) over all frequency bands (ICCs > 0.75), except for delta (ICCs < 0.75), with alpha exhibiting the highest repeatability performance. In addition, females showed reduced reliability compared to males in both PPI and PPF, possibly attributed to menstrual cycle phase across our female participants. Overall, our findings suggest that brain oscillatory activity can be test-retest reliable, while gender needs to be controlled with caution. Finally, event-related EEG oscillations during both PPI and PPF could provide a complementary tool to study psychopathology in clinical practice.

Statistical measure of complexity and correlated behavior of Fermi systems.

We apply the statistical measure of complexity, introduced by López-Ruiz, Mancini, and Calbet (LMC), to uniform Fermi systems. We investigate the connection between information and complexity measures with the strongly correlated behavior of various Fermi systems as nuclear matter, electron gas, and liquid helium. We examine the possibility that LMC complexity can serve as an index quantifying correlations in the specific system and to which extent could be related with experimental quantities. Moreover, we concentrate on thermal effects on the complexity of ideal Fermi systems. We find that complexity behaves, both at low and high values of temperature, in a similar way as the specific heat.

Information entropy, information distances, and complexity in atoms.

Shannon information entropies in position and momentum spaces and their sum are calculated as functions of Z(2 < or = Z < or = 54) in atoms. Roothaan-Hartree-Fock electron wave functions are used. The universal property S = a + b ln Z is verified. In addition, we calculate the Kullback-Leibler relative entropy, the Jensen-Shannon divergence, Onicescu's information energy, and a complexity measure recently proposed. Shell effects at closed-shell atoms are observed. The complexity measure shows local minima at the closed-shell atoms indicating that for the above atoms complexity decreases with respect to neighboring atoms. It is seen that complexity fluctuates around an average value, indicating that the atom cannot grow in complexity as Z increases. Onicescu's information energy is correlated with the ionization potential. Kullback distance and Jensen-Shannon distance are employed to compare Roothaan-Hartree-Fock density distributions with other densities of previous works.

Streptococcus pyogenes clinical isolates and lipoteichoic acid.

Minimally subcultured clinical isolates of virulent nephritogenic and nonnephritogenic Streptococcus pyogenes of the same serotype showed major differences in lipoteichoic acid (LTA) production, secretion, and structure. These were related to changes in coccal adherence to and destruction of growing human skin cell monolayers in vitro. A possible relationship between cellular LTA content and group A streptococcal surface hydrophobicity was also investigated. Nephritogenic S. pyogenes M18 produced twice as much total (i.e., cellular and secretory) LTA as did the virulent, serologically identical, but nonnephritogenic isolate. Also, the LTAs from these organisms differed markedly. The polyglycerol phosphate chain of LTA from the nephritogenic isolate was longer (1.6 times) than was that from the nonnephritogenic isolate. Likewise, both LTAs indicated the presence of alanine and the absence of glucose. Amino sugars were found in LTA from only nephritogenic S. pyogenes. Teichoic acid, as a cellular component or secretory product, was not detected. The adherence of two different nephritogenic group A streptococcal serotypes (M18 and M2) exceeded that of the serologically identical but nonnephritogenic isolates (by about five times), indicating a correlation between virulent strains causing acute glomerulonephritis and adherence to human skin cell monolayers. Likewise, LTA from nephritogenic S. pyogenes M18 was more cytotoxic (1.5 times) than was that from the nonnephritogenic isolate for human skin cells, as determined by protein release. This difference was not perceptible by the more sensitive dye exclusion method (i.e., requiring less LTA), which emphasizes changes in host cell morphology and death. Also, the secretion of LTA by only virulent nephritogenic S. pyogenes M18 was exacerbated by penicillin (a maximum of four times). Finally, while the adherence of nephritogenic S. pyogenes M18 decreased markedly after continued subculturing in vitro, the surface hydrophobicity did not.

[Granulomatous cheilitis. Case report]. / Kokkiomatodes cheilitida. (Anaphora periptoseos)

A case of granulomatous cheilitis, an uncommon disease that is difficult to be treated is described. The disease is of unknown etiology and it may represents a monosymptomatic form of Melkersson-Rosenthal syndrome. The significance of the clinical lesions, the microscopic features as well as the recent aetiological aspects of the disease are discussed.

Differences in penicillin-binding proteins of Streptococcus pyogenes and two derived, stabilized L forms.

The penicillin-binding proteins (PBPs) of Streptococcus pyogenes and two of its derived, stabilized (i.e., nonreverting) L forms, an osmotically fragile L form and a physiologic isotonic L form, were compared. The numbers of PBPs in the membranes of these organisms were 6, 4, and 2 for the coccus and the osmotically fragile and physiologic isotonic L forms, respectively. Likewise, the relative amounts of total PBPs were 1.00: 1.48:0.32 for this coccus and the osmotically fragile and physiologic isotonic L forms, respectively. The two largest PBPs (PBPs 1 and 2) of the coccus were absent in both L forms, while the smallest PBPs (PBPs 5 and 6) were found in all three membranes. Deacylation (half-life) of three of the four PBPs in the osmotically fragile L form membrane required a significantly longer time than did deacylation of these presumed identical enzymes in the parental coccal membrane. Conversely, there was no such difference between the only two PBPs of the physiologic isotonic L form and the same coccal membrane proteins. Intact cells of all three organisms secreted PBPs and what appeared to be penicilloic acid and a minimal amount of free penicillin. A greater amount of these PBPs was secreted by both L forms than by the coccus. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns and ratios of secreted PBPs were identical to those from labeled membrane preparations. These differences are correlated with some of our previous findings and are discussed in terms of inhibition of cell wall synthesis and resulting membrane changes in these two derived, stabilized coccal L forms.

Adherence of Streptococcus agalactiae to synchronously growing human cell monolayers without lipoteichoic acid involvement.

Freshly isolated virulent and nonvirulent strains of Streptococcus agalactiae type III were used to study differences in coccal adherence to synchronously dividing, subconfluent human embryonic amnion and fetal lung monolayers in vitro. The adherence frequency by virulent isolates of mid-logarithmically growing cocci to amnion cells varied markedly with host cell age, being highest shortly after eucaryotic cell division. This variation was not observed with lung cell monolayers, suggesting that cyclic production or exposure of coccal receptor sites on the eucaryotic cell surface with age is not a common property of all primary human cells in vitro. However, and regardless of age, not all cells within these synchronously dividing populations bound virulent cocci, indicating that a very small segment of a population may always be unresponsive to host cell interactions with a coccal pathogen. By comparison, adherence of nonvirulent coccal isolates to amnion and lung cells remained constant and of a very low order, regardless of host cell age. Maximal adherence of virulent S. agalactiae to young host cells occurred at early and mid-logarithmic phases of growth. However, at the late stationary growth phase, adherence was reduced to almost that of nonvirulent isolates. Pretreatment of virulent S. agalactiae with anti-lipoteichoic acid (LTA) serum failed to inhibit coccal adherence to these different host cells. Heat negated adherence. Group B coccal LTA was cytotoxic for these host cells. However, pretreatment of amnion and lung cells with nontoxic levels of this amphiphile did not prevent attachment of virulent cocci. Finally, coccal pretreatment with pronase abrogated adherence to either host cell even though surface-exposed LTA was uneffected, as observed by the indirect fluorescent-antibody procedure. Likewise, no observable difference in surface LTA was detected when fresh isolates of virulent and nonvirulent coccal strains were compared by this procedure. These studies suggest that protein involvement, rather than LTA, is primarily responsible for mediating virulent S. agalactiae type III attachment to these synchronously growing, subconfluent eucaryotic monolayers in vitro.

An electron microscope study of kidney basement membrane changes in the mouse by lipoteichoic acid from Streptococcus pyogenes.

Mice injected repeatedly, intraperitoneally or intravenously, for approximately 1 month with a total of 1.04 mg lipoteichoic acid from a nephritogenic strain of Streptococcus pyogenes lost weight. Analysis by electron microscopy revealed that they also exhibited extensive kidney changes in basement membrane morphology which resembled, in part, those observed in human poststreptococcal glomerulonephritis. For example, the glomerular basement membrane became electron dense and exhibited at least a twofold increase in width sporadically within the same preparation after exposure to lipoteichoic acid. Also, whereas appreciable loss or reduction in epithelial foot processes as a result of fusion was clearly evident, epithelial slits and slit membranes or diaphragms between normal foot processes were not selectively affected. In addition, another mostly thickened, highly coiled or serpentinelike basement membrane with amorphous nodules appeared in these preparations. This type membrane was not observed surrounding the capillary lumina and was the most pronounced abnormality apparent in almost all preparations from mice exposed to lipoteichoic acid. Likewise, the proximal tubular basement membrane became variable in width and increased in electron density in mice given lipoteichoic acid as compared with controls. In addition, this membrane was often punctuated with various morphological protrusions originating from only its thickened areas and which extended away from, and not into, the capillary space. This change was only associated with the basement membrane of the proximal tubular capillaries. All membrane changes persisted but gradually subsided, with normal kidney membrane morphology reappearing on the 4th day following the last injection of lipoteichoic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of streptococcal lipoteichoic acid on prolyl hydroxylase activity as related to collagen formation in mouse fibroblast monolayers.

Dried and wet mouse fibroblast monolayers with labeled collagenous substrate were used to study the effects of lipoteichoic acid (LTA) on cellular prolyl hydroxylase activity. LTA is a scavenger of cations, and Fe2+ is essential for prolyl hydroxylase activity. Surprisingly, addition of LTA to dried monolayers resulted in increased prolyl hydroxylase activity, whereas preincubation of Fe2+ with LTA only negated this increase. However, significant inhibition of enzyme activity by wet monolayers occurred whether LTA was added directly to the test system or whether it was used after preincubation with Fe2+. These data suggest that LTA causes membrane perturbations. Also, that the binding of LTA to the membrane of dried and wet monolayers appears to be decidedly different when based on the subsequent availability of Fe2+ for cellular prolyl hydroxylase activity. The ability of LTA to act as a cationic exchanger and the presence of intracellular Fe2+ inaccessible to LTA probably accounted for the lack of complete inhibition of prolyl hydroxylase activity by this amphiphile in the wet cell system. Considerably less iron was needed to negate the partial inhibition of prolyl hydroxylase activity by LTA in viable cells than was needed to restore the increased enzyme activity by this amphiphile in equivalent dried preparations. These and other results showed that, although LTA does not affect collagen polypeptide chain formation in wet monolayers, its involvement at the molecular level does result in a marked decrease in the hydroxylation of collagenous peptidyl prolyl residues through LTA interaction with Fe2+. This reduction in prolyl hydroxylase activity equaled the reduction in hydroxylation of collagenous protein in fibroblast monolayers caused by LTA reported earlier (O. Leon and C. Panos, Infect. Immun. 40:785-794, 1983). Therefore, these data suggest that partial inhibition of prolyl hydroxylase activity is directly related to the synthesis of defective collagen by wet fibroblast monolayers exposed to minute amounts of group A, type 12 streptococcal LTA. Use of LTA also showed that complete inhibition of hydroxyproline formation is not required for the continued formation and accumulation of defective collagenous protein by these monolayers.

Teichoic acids of Streptococcus agalactiae: chemistry, cytotoxicity, and effect on bacterial adherence to human cells in tissue culture.

The ratio of teichoic acid to lipoteichoic acid (LTA) in a strain of Streptococcus agalactiae type III was found to be 8:1, with the total amount of LTA being 0.1% of the dry weight of the organism. Purified teichoic acid contained D-alanine and possibly a small amount of D-glucose and was approximately 22 glycerol phosphate units in length. The linkage between each of these units was 1-3. In addition, LTA contained a complex lipid, more glucose, and an unusually high content of a short-chain fatty acid, tridecanoic acid. This LTA was cytotoxic for a variety of human cell monolayers in tissue culture, including one derived from the human central nervous system. Established human cells were more sensitive than primary cell monolayers to this LTA, with as little as 12.5 micrograms of LTA per ml being cytotoxic for HeLa cells. Teichoic acid (250 micrograms/ml) was nontoxic under identical conditions. These cytotoxicity results suggest an LTA involvement in group B streptococcal pathogenesis. Also, the first model system for the study of group B streptococcal adherence to primary human embryonic amnion cells in tissue culture is detailed. This system was used to quantitate pronounced differences in tissue tropism between S. agalactiae and Streptococcus pyogenes and showed enhanced binding by this group A coccus over that of S. agalactiae for amnion cell monolayers. The adherence of both streptococcal species to only a portion (40%) of these amnion cells suggested that host cell receptor expression may vary for primary cells in vitro. Finally, this strain of S. agalactiae was shown to adhere to amnion cells by a non-LTA-mediated mechanism. The possibility of an LTA-mediated versus a protein-mediated adherence mechanism for host cells that is related to the virulence of S. agalactiae is discussed.

Morphological changes and pathology of mouse glomeruli infected with a streptococcal L-form or exposed to lipoteichoic acid.

The morphology and pathology of cultured mouse glomeruli were examined at the cellular and subcellular levels after infection with a physiological isotonic L-form of Streptococcus pyogenes type 12 or exposure to streptococcal lipoteichoic acid. These changes, as viewed by light microscopy, were identical regardless of the method used to induce glomerular cytotoxicity. They were characterized by an initial reduction in the outgrowth of cells, some cellular granulation, and later, destruction of the confluent monolayer. Once initiated, cytotoxicity could not be reversed by refeedings, and complete glomerular destruction resulted after 2 weeks. Electron microscope studies revealed that the basement membrane of intact glomeruli exposed to streptococcal lipoteichoic acid had become greatly thickened (two- to fourfold) and electron dense. Our recent biochemical findings have shown that streptococcal lipoteichoic acid increases the amount of collagen formed and retained by mouse fibroblasts in tissue culture as well as causing a reduction in the hydroxylation of proline in both intracellular and secreted collagenous material (Leon and Panos, Infect. Immun. 40:785-794, 1983). These results, together with the present findings, suggest that the thickening of the glomerular basement membrane may be due to defective collagen biosynthesis as a result of streptococcal lipoteichoic acid. The use of cultured glomeruli as a model system for studying the earliest basement membrane alterations in the absence of an immune response as a result of streptococcal lipoteichoic acid is suggested.

Mechanism of inducer expulsion in Streptococcus pyogenes: a two-step process activated by ATP.

The mechanism of methyl-beta-D-thiogalactoside-phosphate (TMG-P) expulsion from Streptococcus pyogenes was studied. The expulsion elicited by glucose was not due to exchange vectorial transphosphorylation between the expelled TMG and the incoming glucose since more beta-galactoside was displaced than glucose taken up, and the stoichiometry between TMG and glucose transport was inconstant. Instead, two distinct and sequential reactions, intracellular dephosphorylation of TMG-P followed by efflux of free TMG, mediated the expulsion. This was shown by temporary accumulation of free TMG effected by competitive inhibition of its efflux and by the aid of arsenate, which arrested dephosphorylation of TMG-P but did not affect efflux of free TMG formed intracellularly before arsenate addition. The competitive inhibition of TMG efflux by its structural analogs suggests that a transport protein facilitates the expulsion. Iodoacetate or fluoride prevented TMG-P dephosphorylation and its expulsion. However, provision of ATP via the arginine deiminase pathway restored these activities in the presence of the glycolytic inhibitors and stimulated expulsion in their absence. Other amino acids tested did not promote this restoration, and canavanine or norvaline severely inhibited it. Arginine without glucose neither elicited the dephosphorylation nor evoked the expulsion of TMG-P. Ionophores or ATPase inhibitors did not prevent the expulsion as elicited by glucose or its restoration by arginine. The results suggest that activation of the dephosphorylation-expulsion mechanism occurs independently of a functional glycolytic pathway, requires ATP provision, and is possibly due to protein phosphorylation controlled by a yet unknown metabolite. The in vivo phosphorylation of a protein (approximate molecular weight - 10,000) under the conditions of expulsion was demonstrated.

Synthesis of 3,4-dihydro-4-oxoquinazoline derivatives as potential anticonvulsants.

Twenty-three substituted 3,4-dihydro-4-oxoquinazolines or 3,4-dihydro-4-oxoazaquinazolines have been synthesized utilizing 2-amino-3-cyano-4,5-dimethylfuran and methyl acrylate as precursors for synthesis of the required substituted anthranilates. Six additional azaquinazolones were synthesized from 2-aminonicotinic or 3-aminopicolinic acid for comparison studies. All compounds were evaluated in mice with the maximal electroshock (MES) seizure and pentylenetetrazol (sc Met) seizure threshold tests for potential anticonvulsant activity and in the rotorod test to evaluate neurotoxicity. Nine of the twenty-nine compounds in the series demonstrated anticonvulsant action. The azaquinazolones were found to possess the most significant activity.

Cytotoxicity and inhibition of normal collagen synthesis in mouse fibroblasts by lipoteichoic acid from Streptococcus pyogenes type 12.

The toxicity of lipoteichoic acid (LTA) from Streptococcus pyogenes type 12 was investigated by using mouse fibroblasts in culture in the absence of serum. Morphologically, while low concentrations of LTA elicited a subtle effect characterized by progressive cellular degeneration with practically no release of protein, larger concentrations (greater than 50 micrograms/ml) of this amphiphile resulted in rapid death of cell monolayers. Metabolic studies utilized a concentration of LTA (17.5 micrograms/ml) which caused the smallest change in cell morphology in the least number of mouse fibroblast cells per monolayer. Under these conditions, cell monolayers showed an increase of 450% in their content of collagenous protein after exposure to LTA. However, the amount of such material secreted remained unchanged. Also, changes in the type of collagenous protein formed were observed after exposure to LTA. Collagenous protein accumulating intracellularly was found to be practically hydroxyproline-free. However, collagenous protein secreted by this cell line showed a significantly reduced content of hydroxyproline as compared with control cells unexposed to this coccal membrane component. Column chromatographic studies confirmed that the collagenous protein secreted by monolayers exposed to LTA was defective (under hydroxylated). It was concluded that LTA does not affect the amount of collagenous protein secreted. However, it does increase the amount of this protein formed and retained by this cell line as well as causing a reduction in the hydroxylation of proline in both intracellular and secreted collagenous material. A possible relationship between abnormal basement membrane morphology and disturbed collagen synthesis in post-streptococcal glomerulonephritis as related to LTA is discussed.

Transport of alpha-aminoisobutyric acid by Streptococcus pyogenes and its derived L-form.

We studied the uptake of alpha-aminoisobutyric acid (AIB) in Streptococcus pyogenes and its physiologically isotonic L-form. S. pyogenes cells starved for glucose or treated with carbonyl cyanide-m-chlorophenyl hydrazone accumulated limited amounts of AIB. A high apparent K(m) value characterized the glucose-independent transport of AIB. The rate and extent of AIB accumulation significantly increased in the presence of glucose. Two saturable transport components with distinct apparent K(m) values characterized glycolysis-coupled transport of AIB. A biphasic Lineweaver-Burk plot was also obtained for l-alanine transport by glycolyzing S. pyogenes cells. AIB seems to share a common transport system(s) with glycine, l- and d-alanine, l-serine, and l-valine. This was shown by the competitive inhibition of AIB uptake by these compounds and their ability to induce competitive exchange efflux of accumulated AIB. About 30% of the AIB uptake was not inhibited by a saturating amount of l-valine, indicating the existence of more than one system for AIB transport. p-Chloromercuribenzoate markedly inhibited the accumulation of AIB by both glycolyzing and glucose-starved cells. In contrast, carbonyl cyanide-m-chlorophenyl hydrazone affected only metabolism-dependent uptake of AIB, which was also sensitive to dinitrophenol, N-ethylmaleimide, iodoacetate, fluoride (NaF), arsenate, and N,N'-dicyclohexylcarbodiimide. These results are interpreted according to the chemiosmotic theory of Mitchell, whereby a proton motive force constitutes the driving force for AIB accumulation. AIB was not accumulated by the L-form. However, a temporary accumulation of AIB by a counterflow mechanism and a saturable system with a low apparent affinity were demonstrated for AIB transport by this organism. We suggest that a deficiency in the coupling of energy to AIB transport is responsible for the apparent lack of active AIB accumulation by the L-form.

Long-chain fatty acid perturbations in Mycoplasma pneumoniae.

The fatty acid content of Mycoplasma pneumoniae increased 2.5- to 9.6-fold when the growth medium was supplemented with a saturated, unsaturated, or beta-hydroxy fatty acid, the greatest increase occurring with palmitic acid. The amount of each supplemented fatty acid found within this organism was 2.8 to 5.5% of the total fatty acid content; the exception was palmitic acid. Up to 57% of the palmitic acid was utilized from the supplemented medium, whereas only 0.2 to 10% of the other fatty acids was utilized. Chromatographic and isotopic analyses revealed that 22% of the labeled palmitic acid incorporated from the palmitic acid-supplemented medium remained free in this organism. Also, even though complex lipid synthesis increased a minimum of 3.8-fold under these conditions, this mycoplasma continued to incorporate intact complex lipids from the growth medium. Bacteriostatic and bactericidal studies which used high concentrations of various long-chain fatty acids showed that only palmitic, myristic, and beta-hydroxydecanoic acids were not bactericidal. The addition of palmitic acid to the growth medium resulted in the formation of exceedingly long, filamentous cells in approximately 25% of the population. Osmotic fragility and electron spin resonance spectroscopy studies showed a correlation among this increased fatty acid content, decreased membrane fluidity, and the increased osmotic fragility of palmitic acid-grown cells. In addition, these cells had a lowered cholesterol content. The effect of such compositional changes on osmotic fragility is discussed in this paper. Finally, the profound increase in the total fatty acid content of palmitic acid-grown cells altered neither sensitivity to tetracycline or erythromycin nor the amount of hydrogen peroxide secreted.