Identifying canonical and replicable multi-scale intrinsic connectivity networks in 100k+ resting-state fMRI datasets.

Despite the known benefits of data-driven approaches, the lack of approaches for identifying functional neuroimaging patterns that capture both individual variations and inter-subject correspondence limits the clinical utility of rsfMRI and its application to single-subject analyses. Here, using rsfMRI data from over 100k individuals across private and public datasets, we identify replicable multi-spatial-scale canonical intrinsic connectivity network (ICN) templates via the use of multi-model-order independent component analysis (ICA). We also study the feasibility of estimating subject-specific ICNs via spatially constrained ICA. The results show that the subject-level ICN estimations vary as a function of the ICN itself, the data length, and the spatial resolution. In general, large-scale ICNs require less data to achieve specific levels of (within- and between-subject) spatial similarity with their templates. Importantly, increasing data length can reduce an ICN's subject-level specificity, suggesting longer scans may not always be desirable. We also find a positive linear relationship between data length and spatial smoothness (possibly due to averaging over intrinsic dynamics), suggesting studies examining optimized data length should consider spatial smoothness. Finally, consistency in spatial similarity between ICNs estimated using the full data and subsets across different data lengths suggests lower within-subject spatial similarity in shorter data is not wholly defined by lower reliability in ICN estimates, but may be an indication of meaningful brain dynamics which average out as data length increases.

Acute objective and subjective intoxication effects of legal-market high potency THC-dominant versus CBD-dominant cannabis concentrates.

As the market for cannabis concentrate products grows, the lack of research regarding the effects of concentrated THC and CBD becomes more glaring. The present study analyzes cannabinoid blood levels and subjective outcomes of physical sensation and affective state after ad libitum use of legal-market concentrate products. Recreational cannabis users were randomly assigned to THC- or CBD-dominant concentrate products, completing a baseline session, and an experimental mobile laboratory session consisting of timepoints before, immediately after, and one-hour after concentrate use. THC-dominant concentrates induced higher intoxication, and higher ratings of drug effect and drug liking than the CBD-dominant concentrate. Both products induced immediate feelings of elation, diminishing over the subsequent hour. Subjective outcomes in the CBD-dominant group revealed immediate decreases in tension and anxiety relative to pre-use, while the THC-dominant group only saw significant decreases in anxiety after one hour. Paranoia spiked immediately post-use in THC-dominant concentrate users, returning to baseline within an hour. Overall, the CBD-dominant concentrate invoked positive mood effects, lower intoxication and an absence of undesirable effects experienced with the THC-dominant concentrate, potentially mitigating negative effects when combined. Results support the need for further investigation into harm-reduction potential of concentrated CBD when used alone and with THC.

THC and CBD effects on alcohol use among alcohol and cannabis co-users.

OBJECTIVE: Conflicting evidence exists regarding the effects of cannabis on alcohol consumption, with some studies suggesting that cannabis is a substitute for alcohol, whereas others suggest that cannabis complements alcohol, thereby increasing drinking. Cannabidiol (CBD) has shown preclinical promise in decreasing alcohol consumption. This study explores the effects of cannabis containing different potencies of CBD and delta-9-tetrahydrocannabinol (THC) on alcohol consumption. METHOD: In this naturalistic observational study, 120 cannabis and alcohol-using adults (mean age = 33.2 years, 39.2% female, 83.3% white) were assigned to use one of three legal-market cannabis strains (predominantly THC, predominantly CBD, and CBD + THC) ad libitum for 5 days. Timeline Followback data on drinking and cannabis use were collected at a baseline session pertaining to the 30 days prior to the ad libitum period, and data regarding alcohol and cannabis use during the 5-day period were collected at follow-up (FU), immediately following the 5-day period. RESULTS: Regression models tested strain differences in drinking outcomes during the ad libitum period. Orthogonal contrast codes were created comparing the CBD group with the other two groups and comparing the THC group with the CBD + THC group. The CBD group drank fewer drinks per drinking day (p < .05), had fewer alcohol use days (p < .05), and fewer alcohol and cannabis co-use days (p < .05) compared with the other groups. No differences emerged between the THC and the CBD + THC group. CONCLUSIONS: Cannabinoid content should be considered in studies of alcohol and cannabis co-use. Findings are consistent with preclinical work, suggesting that CBD may be associated with decreased alcohol consumption. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

The CRHR1 gene, trauma exposure, and alcoholism risk: a test of G × E effects.

The corticotropin-releasing hormone type I receptor (CRHR1) gene has been implicated in the liability for neuropsychiatric disorders, particularly under conditions of stress. On the basis of the hypothesized effects of CRHR1 variation on stress reactivity, measures of adulthood traumatic stress exposure were analyzed for their interaction with CRHR1 haplotypes and single-nucleotide polymorphisms (SNPs) in predicting the risk for alcoholism. Phenotypic data on 2533 non-related Caucasian individuals (1167 alcoholics and 1366 controls) were culled from the publically available Study of Addiction: Genetics and Environment genome-wide association study. Genotypes were available for 19 tag SNPs. Logistic regression models examined the interaction between CRHR1 haplotypes/SNPs and adulthood traumatic stress exposure in predicting alcoholism risk. Two haplotype blocks spanned CRHR1. Haplotype analyses identified one haplotype in the proximal block 1 (P = 0.029) and two haplotypes in the distal block 2 (P = 0.026, 0.042) that showed nominally significant (corrected P < 0.025) genotype × traumatic stress interactive effects on the likelihood of developing alcoholism. The block 1 haplotype effect was driven by SNPs rs110402 (P = 0.019) and rs242924 (P = 0.019). In block 2, rs17689966 (P = 0.018) showed significant and rs173365 (P = 0.026) showed nominally significant, gene × environment (G × E) effects on alcoholism status. This study extends the literature on the interplay between CRHR1 variation and alcoholism, in the context of exposure to traumatic stress. These findings are consistent with the hypothesized role of the extra hypothalamic corticotropin-releasing factor system dysregulation in the initiation and maintenance of alcoholism. Molecular and experimental studies are needed to more fully understand the mechanisms of risk and protection conferred by genetic variation at the identified loci.

Brain-derived neurotrophic factor genotype is associated with brain gray and white matter tissue volumes recovery in abstinent alcohol-dependent individuals.

Neuroimaging studies have linked the methionine (Met) allele of the brain-derived neurotrophic factor (BDNF) gene to abnormal regional brain volumes in several psychiatric and neurodegenerative diseases. However, no neuroimaging studies assessed the effects of this allele on brain morphology in alcohol use disorders and its demonstrated change during abstinence from alcohol. Here we assessed the effects of the BDNF Val66Met (rs6265) polymorphism on regional brain tissue volumes and their recovery during short-term abstinence in treatment-seeking alcohol-dependent individuals. 3D T1 weighted magnetic resonance images from 62 individuals were acquired at 1.5 T at one week of abstinence from alcohol; 41 of the participants were rescanned at 5 weeks of abstinence. The images were segmented into gray matter (GM), white matter (WM) and cerebrospinal fluid and parcellated into regional volumes. The BDNF genotype was determined from blood samples using the TaqMan technique. Alcohol-dependent Val (Valine)/Met heterozygotes and Val homozygotes had similar regional brain volumes at either time point. However, Val homozygotes had significant GM volume increases, while Val/Met heterozygotes increased predominantly in WM volumes over the scan interval. Longitudinal increases in GM but not WM volumes were related to improvements in neurocognitive measures during abstinence. The findings suggest that functionally significant brain tissue volume recovery during abstinence from alcohol is influenced by BDNF genotype.

The role of genomics in health behavior change: challenges and opportunities.

The most common causes of morbidity and mortality in the western world can be accounted for by unhealthy patterns of behavior (e.g. smoking, sedentary lifestyle, unhealthy diet, and alcohol consumption). Interventions to improve health behavior are sorely needed. To fully realize the potential of health behavior change interventions, be they individual level, community level, social structural, or policy-based, a greater understanding of the extent to which genomics can inform efforts at health behavior change is warranted. In this commentary, we explore three relatively novel possible routes to the integration of genomics and health behavior: (1) genomics may influence health behavior indirectly through intermediate phenotypes, requiring well-defined theory-based and mechanistic models of health behavior, (2) genomics may moderate response to interventions to change health behavior, and (3) genomics, specifically epigenetic variation, may be influenced by health behavior. The integration of genomics into research on interventions to change health behavior is not without challenges and will certainly require transdisciplinary collaborative science to succeed. We provide specific action points for moving the science forward to explore the extent to which genomic information can be harnessed to ultimately decrease morbidity and mortality associated with unhealthy behavior.

CHRNB2 promoter region: association with subjective effects to nicotine and gene expression differences.

Smoking behavior is a complex, which includes multiple stages in the progression from experimentation to continued use and dependence. The experience of subjective effects, such as dizziness, euphoria, heart pounding, nausea and high, have been associated with varying degrees of persistence and subsequent abuse/dependence of marijuana, cocaine, tobacco and alcohol (Grant et al. 2005, Wagner & Anthony 2002). Previous studies have reported associations between neuronal nicotinic receptor (CHRN) genes and subjective effects to nicotine. We sought to replicate and expand this work by examining eight single nucleotide polymorphisms (SNPs) in a sample of adult smokers (n = 316) who reported subjective effects following cigarette smoking in a controlled laboratory environment. Two SNPs each in the CHRNB2, CHRNB3, CHRNA6 and CHRNA4 genes were examined. A significant association was found between two SNPs and physical effects reported after smoking the first experimental cigarette. SNP rs2072658 is upstream of CHRNB2 (P-value = 0.0046) and rs2229959 is a synonymous change in exon 5 of CHRNA4 (P value = 0.0051). We also examined possible functional relevance of SNP rs2072658 using an in vitro gene expression assay. These studies provided evidence that the minor allele of rs2072658 may lead to decreased gene expression, using two separate cell lines, P19 and SH-SY5Y (18% P < 0.001 and 26% P < 0.001 respectively). The human genetic study and functional assays suggest that variation in the promoter region of CHRNB2 gene may be important in mediating levels of expression of the ß2 nicotinic receptor subunit, which may be associated with variation in subjective response to nicotine.

Genetic contributions to avoidance-based decisions: striatal D2 receptor polymorphisms.

Individuals differ in their tendencies to seek positive decision outcomes or to avoid negative ones. At the neurobiological level, our model suggests that phasic changes in dopamine support learning to reinforce good decisions via striatal D1 receptors, and to avoid maladaptive choices via striatal D2 receptors. Accordingly, in a previous study individual differences in positive and negative learning were strongly modulated by two genetic polymorphisms factors related to striatal D1 and D2 function, respectively. Nevertheless, whereas the role for dopamine in positive learning is relatively well accepted, that in learning to avoid negative outcomes is more controversial. Here we further explore D2-receptor-related genetic contributions to probabilistic avoidance in humans, in light of recent data showing that particular DRD2 polymorphisms are associated with functional modulation of receptor expression [Zhang Y, Bertolino A, Fazio L, Blasi G, Rampino A, Romano R, Lee M-LT, Xiao T, Papp A, Wang D, Sadée W (2007) Polymorphisms in human dopamine d2 receptor gene affect gene expression, splicing, and neuronal activity during working memory. Proc Natl Acad Sci U S A 104(51):20552-20557]. We find that a promoter polymorphism rs12364283 associated with transcription and D2 receptor density was strongly and selectively predictive of avoidance-based decisions. Two further polymorphisms (rs2283265 and rs1076560) associated with relatively reduced presynaptic relative to postsynaptic D2 receptor expression were predictive of relative impairments in negative compared to positive decisions. These previously undocumented effects of DRD2 polymorphisms were largely independent of those we reported previously for the C957T polymorphism (rs6277) associated with striatal D2 density. In contrast, effects of the commonly studied Taq1A polymorphism on reinforcement-based decisions were due to indirect association with C957T. Taken together these findings suggest multiple D2-dependent genetic mechanisms contributing to avoidance. We discuss these effects in the context of neurocomputational models of reinforcement leaning in the basal ganglia.

Human gamma-aminobutyric acid A receptor alpha2 gene moderates the acute effects of alcohol and brain mRNA expression.

Gamma-aminobutyric acid A (GABA(A)) receptors moderate several of the behavioral effects of alcohol. In fact, recent studies have shown an association between the gene for the alpha2-subunit of the GABA(A) receptor (GABRA2) and alcoholism. In the present study, we examined the functional relevance of the GABRA2 gene in alcohol dependence by assessing brain GABRA2 mRNA and GABA(A)alpha2-subunit protein levels in post-mortem prefrontal cortical tissue collected from control and alcohol-dependent individuals. In addition, using an endophenotype approach, we tested whether the GABRA2 gene moderates sensitivity to the acute effects of alcohol in two independent samples from distinct human alcohol challenge studies. Results indicated that GABRA2 mRNA levels significantly differed by GABRA2 genotype. GABRA2 single nucleotide polymorphisms (rs573400, rs279871 and rs279858) were significantly associated with sensitivity to the acute effects of alcohol. Specifically, there was a significant main effect of GABRA2 x breath alcohol concentration on several measures of subjective responses to alcohol, including the hedonic value of alcohol. Importantly, reanalysis of a previous intravenous alcohol administration study confirmed the results of the oral alcohol challenge study. In summary, these results extend previous findings and provide new insights into the putative biobehavioral mechanisms that may moderate the association between the GABRA2 gene, sensitivity to the acute effects of alcohol and ultimately alcohol dependence.

Genome plasticity in the mouse oocyte and early embryo.

In dissecting the molecules and molecular mechanisms that control mammalian oocyte-to-embryo transition, we found abundant transcripts representing developmentally regulated ERVs (endogenous retroviruses) in mouse oocyte and two-cell stage embryo cDNA libraries. These retrotransposons can act as alternative promoters and first exons for diverse genes, synchronizing their expression. Heritable genetic change due to replication of these retrotransposons probably occurs specifically in oocytes and early embryos. ERVs are usually epigenetically silenced, through DNA methylation and chromatin-based mechanisms. Their activation and silencing indicates a change in the epigenetic state of the genome. The thousands of endogenous retro-elements in the mouse genome provides potential scope for large-scale co-ordinated epigenetic fluctuations and leads to the hypothesis that differential transposable element expression triggers sequential reprogramming of the embryonic genome during the oocyte-to-embryo transition.

Comparison of the University of California at Los Angeles Line-by-Line Equivalent Radiative Transfer Model and the Moderate-Resolution Transmission Model for accuracy assessment of the National Polar-Orbiting Operational Environmental Satellite System's Visible-Infrared Imager-Radiometer Suite cloud algorithms.

To support the verification and implementation of the National Polar-Orbiting Operational Environmental Satellite System's Visible-Infrared Imaging-Radiometric Suite (VIIRS) algorithms used for inferring cloud environmental data records, an intercomparison effort has been carried out to assess the consistency between the simulated cloudy radiances-reflectances from the University of California at Los Angeles Line-by-Line Equivalent Radiative Transfer Model and those from the Moderate-Resolution Transmission Model (MODTRAN) with the 16 stream Discrete Ordinate Radiative Transfer Model (DISORT) incorporated. For typical ice and water cloud optical depths and particle sizes, we found discrepancies in the visible and near-infrared reflectances from the two models, which presumably are due to the difference in phase function (nonspherical versus Henyey-Greenstein), different numbers of phase function expansion terms (16 versus 200 terms), and different treatment of forward peak truncation in each model. Using the MODTRAN4, we also found substantial differences in the infrared radiances for optically thick clouds. These differences led to the discovery by MODTRAN4 developers of an inconsistency in the MODTRAN4-DISORT interface. MODTRAN4 developers corrected the inconsistency, which provided dramatic reductions in the differences between the two radiative transfer models. The comparison not only affects the prospective test plan for the VIIRS cloud algorithms but also should lead to improvements in future MODTRAN releases.

Regional scale flood modeling using NEXRAD rainfall, GIS, and HEC-HMS/RAS: a case study for the San Antonio River Basin Summer 2002 storm event.

This paper develops a framework for regional scale flood modeling that integrates NEXRAD Level III rainfall, GIS, and a hydrological model (HEC-HMS/RAS). The San Antonio River Basin (about 4000 square miles, 10,000 km2) in Central Texas, USA, is the domain of the study because it is a region subject to frequent occurrences of severe flash flooding. A major flood in the summer of 2002 is chosen as a case to examine the modeling framework. The model consists of a rainfall-runoff model (HEC-HMS) that converts precipitation excess to overland flow and channel runoff, as well as a hydraulic model (HEC-RAS) that models unsteady state flow through the river channel network based on the HEC-HMS-derived hydrographs. HEC-HMS is run on a 4 x 4 km grid in the domain, a resolution consistent with the resolution of NEXRAD rainfall taken from the local river authority. Watershed parameters are calibrated manually to produce a good simulation of discharge at 12 subbasins. With the calibrated discharge, HEC-RAS is capable of producing floodplain polygons that are comparable to the satellite imagery. The modeling framework presented in this study incorporates a portion of the recently developed GIS tool named Map to Map that has been created on a local scale and extends it to a regional scale. The results of this research will benefit future modeling efforts by providing a tool for hydrological forecasts of flooding on a regional scale. While designed for the San Antonio River Basin, this regional scale model may be used as a prototype for model applications in other areas of the country.

With great expectations, can two "wrongs" prime a "right"?

The proportion of related prime-target pairs (relatedness proportion, RP) and prime-target stimulus onset asynchrony (SOA) was varied to determine the involvement of strategic priming mechanisms in the reduction in semantic priming that occurs when a target follows an unmasked prime that itself receives immediate repetition priming from a masked prime. At 300-ms and 1,200-ms SOAs, (a) strategic semantic priming was operating, in that priming from a nonrepeated prime increased as RP increased from .25 to .75, and (b) for both RPs, prime repetition reduced semantic priming. At a 167-ms SOA, (a) priming from a nonrepeated prime was unaffected by RP, suggesting that strategic priming was not operating, and (b) for both RPs, prime repetition did not reduce semantic priming. Because prime repetition did not reduce priming at the 167-ms SOA (when only spreading activation should have been mediating semantic priming), the reduction in semantic priming produced by prime repetition is not evidence against spreading activation automaticity. Possible mechanisms through which prime repetition reduces semantic priming are discussed.

Dynamic NMR studies of a potential chiroptical switch based on dithiocarbamate-iminodithiolane interconversion.

[reaction: see text]. Variable temperature NMR spectra of the chiral spiro[(4-N,N-dimethyldithiocarbamato)-(2-N,N-dimethylimino)-1,3-dithiolane-5,9'-xanthene] show complex dynamics including degenerate interconversion of the dithiocarbamate and iminodithiolane groups. The rate of this switching process can be controlled by chemical modification: the analogous spiro[dithiolane-fluorene] derivative shows no interconversion. These novel materials have potential application as molecular switching elements in information storage devices.

Attentional capture by irrelevant color singletons.

Four experiments investigated attentional capture by a color singleton in visual search for a nonsingleton target. B. S. Gibson and E. M. Kelsey (1998) found that a color singleton in a precuing array facilitated target discrimination at that location if the same color also signaled the target array onset. The authors found similar cuing effects regardless of whether the singleton color matched the target array and regardless of whether subjects could anticipate the singleton or target-array color. In Experiment 4, a color singleton captured attention when it appeared in the precuing array but not when it appeared in the target array. The results indicate that attentional control settings for displaywide attributes are imprecise: Although subjects may anticipate a specific color, they cannot avoid attentional capture by other irrelevant colors. In addition, the effect of irrelevant singletons depends on whether a target is simultaneously present in the array.

Bonding of Hg(II) to reduced organic sulfur in humic acid as affected by S/Hg ratio.

Organic matter is an important sorbent of heavy metals in soils and sediments. The heterogeneity of organic matter, including the presence of various reactive O-, N-, and S-bearing ligands, makes it difficult to precisely characterize the nature of metal-ligand binding sites. The objective of this research was to characterize the extent and nature of Hg(II) bonding with reduced organic S in soil organic matter. Sulfur-rich humic acid (0.7 +/- 0.1 mol of S kg-1) was extracted from samples of surface soil from a marine wetland. Synchrotron X-ray absorption near-edge structure (XANES) analysis at the S K edge indicated that 70 +/- 3 mol % of the organic S was in a reduced oxidation state. Aqueous solutions containing 2 mmol of Hg kg-1, 0.1 M NaNO3, and humic acid added at various S/Hg molar ratios at pH 5.60 +/- 0.02 were characterized using extended X-ray absorption fine structure (EXAFS) spectroscopy at the Hg LIII edge. Spectral fitting showed that as the total S/Hg ratio increased from 0.6 to 5.6 (reduced S/Hg of 0.4-4.0), the fraction of Hg-S bonding relative to Hg-O (or Hg-N) bonding increased from 0.4 to 0.9. Results demonstrated preferential bonding of Hg(II) to reduced organic S sites and indicated that multiple sulfur ligands were coordinated with Hg2+ ions at high S/Hg ratios, which corresponded to low levels of complexed Hg(II).

Olanzapine reduces urge to drink after drinking cues and a priming dose of alcohol.

RATIONALE: Haloperidol, a D2 antagonist, has been shown to moderate the effects of alcohol consumption on craving. OBJECTIVE: The present study was designed to determine whether a single 5-mg dose of olanzapine (a D2/5-HT2 antagonist) would influence responses to alcohol cues or an alcohol challenge. It was hypothesized that olanzapine would attenuate cue-elicited urge to drink, attenuate the effects of alcohol consumption on urge to drink, and reduce the rewarding effects of alcohol. METHODS: To test these hypotheses, 26 heavy social drinkers were randomized to receive either 5 mg olanzapine or placebo approximately 8 h before each of two experimental sessions. Participants consumed a moderate dose of alcohol in one experimental session and a non-alcohol control beverage in another session. RESULTS: Results indicated that mere exposure to alcohol cues and consumption of alcohol increased urge to drink and that olanzapine attenuated these effects. Results also indicated that alcohol increased subjective stimulation and high while olanzapine did not moderate these effects. CONCLUSIONS: These results suggest that olanzapine did not influence the rewarding effects of alcohol but did attenuate the effects of alcohol cues and an alcohol challenge on urge to drink.

A content analysis of smoking craving.

The purpose of this study was to conduct a content analysis of smoking craving in order to investigate more precisely the subjective nature of the construct with the goal of informing assessment. Thirty-two smokers interested in cessation treatment provided free response written descriptions of the level of craving they normally experience. These responses were analyzed for subjective content along five theoretical domains: physiological, affective, cognitive, behavioral, and synonyms (of craving). Although there were no differences in the relative proportion of broad content terms smokers used to describe their craving (e.g., cognitive versus affective), this analysis revealed considerable diversity in the specific terms smokers used. Some smokers described their craving in purely physiological terms whereas others used primarily cognitive terms, and still others used affective terms. To assume that smoking craving is qualitatively similar across persons, then, may mask important variations that define the individual experience of craving.

Toward bridging the gap between biological, psychobiological and psychosocial models of alcohol craving.

Urge to drink ("craving") has been a central focus of many theories and treatments, but some researchers question the importance of urges during recovery. Several studies assessed reactions to the presence of beverage alcohol (cue-reactivity) or to simulated high-risk situations (role plays). Higher urges in response to role plays predicted more drinking during the 6 months after treatment. However, urges in response to beverage cues were inconsistently predictive of outcome while measures of awareness or attention to cues predicted less drinking. Urge to drink might reflect a conflict between motivation to drink and awareness of danger. Whether urges predict increased risk of drinking should be a function of factors that affect motivation to drink, awareness of risk and effectiveness of coping. Cue-reactivity assessment has recently been used to bridge the gap between psychosocial and biomedical approaches in several ways: (1) salivation to cues predicts increased drinking independent of urge or attention, showing the value of including both physiological and psychosocial measures; (2) naltrexone has been shown to decrease cue-elicited urge to drink, illustrating the value of this assessment methodology for medications evaluation and (3) pre-pulse inhibition of startle response is being used to investigate the role of dopaminergic pathways in cue-elicited urge. Thus, this laboratory based program of research has the potential to add to knowledge of both biomedical and psychosocial mechanisms involved in urge and relapse, leading to greater integration of models.

Immunomagnetic purification of Colletotrichum lindemuthianum appressoria.

We developed a method to purify appressoria of the bean anthracnose fungus Colletotrichum lindemuthianum for biochemical analysis of the cell surface and to compare appressoria with other fungal structures. We used immunomagnetic separation after incubation of infected bean leaf homogenates with a monoclonal antibody that binds strongly to the appressoria. Preparations with a purity of >90% could be obtained. Examination of the purified appressoria by transmission electron microscopy showed that most had lost their cytoplasm. However, the plasma membrane was retained, suggesting that there is some form of attachment of this membrane to the cell wall. The purified appressoria can be used for studies of their cell surface, and we have shown that there are clear differences in the glycoprotein constituents of cell walls of appressoria compared with mycelium.