NECAB2 is an endosomal protein important for striatal function.

Synaptic signaling depends on ATP generated by mitochondria. Dysfunctional mitochondria shift the redox balance towards a more oxidative environment. Due to extensive connectivity, the striatum is especially vulnerable to mitochondrial dysfunction. We found that neuronal calcium-binding protein 2 (NECAB2) plays a role in striatal function and mitochondrial homeostasis. NECAB2 is a predominantly endosomal striatal protein which partially colocalizes with mitochondria. This colocalization is enhanced by mild oxidative stress. Global knockout of Necab2 in the mouse results in increased superoxide levels, increased DNA oxidation and reduced levels of the antioxidant glutathione which correlates with an altered mitochondrial shape and function. Striatal mitochondria from Necab2 knockout mice are more abundant and smaller and characterized by a reduced spare capacity suggestive of intrinsic uncoupling respectively mitochondrial dysfunction. In line with this, we also found an altered stress-induced interaction of endosomes with mitochondria in Necab2 knockout striatal cultures. The predominance of dysfunctional mitochondria and the pro-oxidative redox milieu correlates with a loss of striatal synapses and behavioral changes characteristic of striatal dysfunction like reduced motivation and altered sensory gating. Together this suggests an involvement of NECAB2 in an endosomal pathway of mitochondrial stress response important for striatal function.

Single-molecule tracking dataset of histone H3 (Hht1) in Saccharomyces cerevisiae.

Single-Molecule Tracking (SMT) is a powerful method to quantify protein dynamics in live cells. Recently, we have established a data analysis pipeline for estimating various biophysical parameters (mean squared displacement, diffusion coefficient, bound fraction, residence time, jump distances, jump angles, and track statistics) from the single-molecule time-lapse movies acquired from yeast Saccharomyces cerevisiae. We acquired the time-lapse movies using different time intervals (i.e. 15 ms, 200 ms, and 1000 ms) to extract the diffusion parameters (from 15 ms time interval movies) and residence time (from 200 ms and 1000 ms time interval movies). We tracked the single molecules from these movies using three MATLAB-based software packages (MatlabTrack, TrackIT, DiaTrack (Sojourner, and Spot-On)) to quantify various biophysical parameters. In this article, we have quantified the biophysical parameters of chromatin-bound histone H3 (Hht1), labeled using JF646 HaloTag Ligand (HTL), and shared a few raw time-lapse SMT movies for the same. Histone H3 is a chromatin-bound protein and it serves as a benchmark for the stably bound molecules for the SMT experiments. Hence, this dataset can be used by various researchers to quantify the biophysical parameters of chromatin-bound molecules (Histone H3). Any newly developed tracking software can use this dataset to validate the accuracy of its tracking algorithms.

Predictors of Severity of Scrub Typhus in Children Requiring Pediatric Intensive Care Admission.

Objective of our study was to determine the clinical characteristics and laboratory profile of scrub typhus patients requiring pediatric intensive care admission and to find out risk factors for the severity of illness. This was a cross-sectional observational study conducted on 1-month to 12-year-old children admitted with scrub typhus in a tertiary care pediatric intensive care unit (PICU). Relevant demographic, clinical, laboratory, treatment, and outcome-related data were documented. The severity of the disease was measured in the form of multiple organ dysfunction syndrome (MODS). With further correlation, and univariate and multivariate analyses, factors associated with severe disease were identified. During the study period, out of 586 PICU admission, 62 patients (10.6%) were diagnosed with scrub typhus. The mean age was 63.85 ± 52.78 months, where infants constituted 32.3% of the total population. Fever was present in 100% of the cases. Common indications of PICU admission were: respiratory distress 42 (67.7%), altered sensorium 41 (66.1%), convulsion 37 (59.7%), and shock 31 (50%). Total number of patients with MODS was 40 (64.5%). The case fatality rate was 8%. On multivariate analysis, infant age group ( p = 0.02), altered sensorium ( p = 0.001), reduced urine output ( p = 0.02), thrombocytopenia ( p = 0.001), raised C-reactive protein ( p = 0.004), hyponatremia ( p = 0.005), hypoalbuminemia ( p = 0.01), deranged international normalized ratio ( p = 0.02), and hyperferritinemia ( p = 0.02) came out to be independent factors in predictability for development of MODS. Multiorgan dysfunction is a life-threatening manifestation of scrub typus in children, which necessitates PICU admission. Infant age group, presence of altered sensorium, reduced urine output, thrombocytopenia, elevated inflammatory markers, coagulopathy, hypoalbuminemia, and hyponatremia predict risk for MODS.

Single-molecule tracking for studying protein dynamics and target-search mechanism in live cells of S. cerevisiae.

Single-molecule tracking (SMT) is a powerful approach to quantify the biophysical parameters of protein dynamics in live cells. Here, we describe a protocol for SMT in live cells of the budding yeast Saccharomyces cerevisiae. We detail how to genetically engineer yeast strains for SMT, how to set up image acquisition parameters, and how different software programs can be used to quantify a variety of biophysical parameters such as diffusion coefficient, residence time, bound fraction, jump angles, and target-search parameters. For complete details on the use and execution of this protocol, please refer to Mehta et al. 1 and Ball et al..2.

Control of protein synthesis and memory by GluN3A-NMDA receptors through inhibition of GIT1/mTORC1 assembly.

De novo protein synthesis is required for synapse modifications underlying stable memory encoding. Yet neurons are highly compartmentalized cells and how protein synthesis can be regulated at the synapse level is unknown. Here, we characterize neuronal signaling complexes formed by the postsynaptic scaffold GIT1, the mechanistic target of rapamycin (mTOR) kinase, and Raptor that couple synaptic stimuli to mTOR-dependent protein synthesis; and identify NMDA receptors containing GluN3A subunits as key negative regulators of GIT1 binding to mTOR. Disruption of GIT1/mTOR complexes by enhancing GluN3A expression or silencing GIT1 inhibits synaptic mTOR activation and restricts the mTOR-dependent translation of specific activity-regulated mRNAs. Conversely, GluN3A removal enables complex formation, potentiates mTOR-dependent protein synthesis, and facilitates the consolidation of associative and spatial memories in mice. The memory enhancement becomes evident with light or spaced training, can be achieved by selectively deleting GluN3A from excitatory neurons during adulthood, and does not compromise other aspects of cognition such as memory flexibility or extinction. Our findings provide mechanistic insight into synaptic translational control and reveal a potentially selective target for cognitive enhancement.

In-vivo Single-Molecule Imaging in Yeast: Applications and Challenges.

Single-molecule imaging has gained momentum to quantify the dynamics of biomolecules in live cells, as it provides direct real-time measurements of various cellular activities under their physiological environment. Yeast, a simple and widely used eukaryote, serves as a good model system to quantify single-molecule dynamics of various cellular processes because of its low genomic and cellular complexities, as well as its facile ability to be genetically manipulated. In the past decade, significant developments have been made regarding the intracellular labeling of biomolecules (proteins, mRNA, fatty acids), the microscopy setups to visualize single-molecules and capture their fast dynamics, and the data analysis pipelines to interpret such dynamics. In this review, we summarize the current state of knowledge for the single-molecule imaging in live yeast cells to provide a ready reference for beginners. We provide a comprehensive table to demonstrate how various labs tailored the imaging regimes and data analysis pipelines to estimate various biophysical parameters for a variety of biological processes. Lastly, we present current challenges and future directions for developing better tools and resources for single-molecule imaging in live yeast cells.

An aspartyl protease-mediated cleavage regulates structure and function of a flavodoxin-like protein and aids oxidative stress survival.

A family of eleven glycosylphosphatidylinositol-anchored aspartyl proteases, commonly referred to as CgYapsins, regulate a myriad of cellular processes in the pathogenic yeast Candida glabrata, but their protein targets are largely unknown. Here, using the immunoprecipitation-mass spectrometry approach, we identify the flavodoxin-like protein (Fld-LP), CgPst2, to be an interactor of one of the aspartyl protease CgYps1. We also report the presence of four Fld-LPs in C. glabrata, which are required for survival in kidneys in the murine model of systemic candidiasis. We further demonstrated that of four Fld-LPs, CgPst2 was solely required for menadione detoxification. CgPst2 was found to form homo-oligomers, and contribute to cellular NADH:quinone oxidoreductase activity. CgYps1 cleaved CgPst2 at the C-terminus, and this cleavage was pivotal to oligomerization, activity and function of CgPst2. The arginine-174 residue in CgPst2 was essential for CgYps1-mediated cleavage, with alanine substitution of the arginine-174 residue also leading to elevated activity and oligomerization of CgPst2. Finally, we demonstrate that menadione treatment led to increased CgPst2 and CgYps1 protein levels, diminished CgYps1-CgPst2 interaction, and enhanced CgPst2 cleavage and activity, thereby implicating CgYps1 in activating CgPst2. Altogether, our findings of proteolytic cleavage as a key regulatory determinant of CgPst2, which belongs to the family of highly conserved, electron-carrier flavodoxin-fold-containing proteins, constituting cellular oxidative stress defense system in diverse organisms, unveil a hidden regulatory layer of environmental stress response mechanisms.

Wire electrical discharge machining and microstructural analysis of hot-pressed boron carbide.

Boron carbide powder was hot-pressed at 2070 °C with 30 MPa uniaxial pressure and 90 min soaking. The mechanical, microstructure and other related properties were evaluated. XRD of the boron carbide powder and sintered samples, shows the presence of B13C2 phase of high electrical conductivity. Crystal lattice parameters, space group, cell angle, cell parameters, etc. were found from Rietveld refinement. The micro Vicker's hardness was 26.98 ± 0.98 GPa at 4.9 N load, fracture toughness 3.54 ± 0.26 MPa m and Young's modulus 461.50 ± 4.5 GPa. The hot-pressed boron carbide was found to be electrically conducting, which can be machined using a wire electrical discharge machine (WEDM).

New Probucol Analogues Inhibit Ferroptosis, Improve Mitochondrial Parameters, and Induce Glutathione Peroxidase in HT22 Cells.

Probucol, a hypocholesterolemic compound, is neuroprotective in several models of neurodegenerative diseases but has serious adverse effects in vivo. We now describe the design and synthesis of two new probucol analogues that protect against glutamate-induced oxidative cell death, also known as ferroptosis, in cultured mouse hippocampal (HT22) cells and in primary cortical neurons, while probucol did not show any protective effect. Treatment with both compounds did not affect glutathione depletion but still significantly decreased glutamate-induced production of oxidants, mitochondrial superoxide generation, and mitochondrial hyperpolarization in HT22 cells. Both compounds increase glutathione peroxidase (GPx) 1 levels and GPx activity, also exhibiting protection against RSL3, a GPx4 inactivator. These two compounds are therefore potent activators of GPx activity making further studies of their neuroprotective activity in vivo worthwhile.

Anomalous kinetic roughening in growth of MoS2 films under pulsed laser deposition.

Growth dynamics of thin films expressed by scaling theory is a useful tool to quantify the statistical properties of the surface morphology of the thin films. To date, the growth mechanism for 2D van der Waals materials has been rarely investigated. In this work, an experimental investigation was carried out to identify the scaling behavior as well as the growth mechanism of 2D MoS2 thin films, grown on glass substrates by pulsed laser deposition for different deposition time durations, using atomic force microscopy images. The growth of MoS2 films evolved from layer-by-layer to layer plus island with the increase in deposition time from 20 s to 15 min. The film surface exhibited anisotropic growth dynamics in the vertical and lateral directions where RMS roughness varied with deposition time as w ∼ t ß with the growth exponent ß = 0.85 ± 0.11, while the lateral correlation length ξ was ξ = t 1/z with 1/z = 0.49 ± 0.09. The films showed a local roughness exponent α loc = 0.89 ± 0.01, global roughness exponent α = 1.72 ± 0.14 and spectral roughness exponent α s = 0.85 ± 0.03, suggesting that the growth of MoS2 thin films followed intrinsic anomalous scaling behavior (α s < 1, α loc = α s ≠ α). Shadowing owing to conical incoming particle flux distribution towards the substrate during deposition has been attributed to the anomalous growth mode. The optical properties of the films, extracted from ellipsometric analysis, were also correlated with RMS roughness and cluster size variation which unveiled the important role played by surface roughness and film density.

Correlation between surface scaling behavior and surface plasmon resonance properties of semitransparent nanostructured Cu thin films deposited via PLD.

The surface scaling behavior of nanostructured Cu thin films, grown on glass substrates by the pulsed laser deposition technique, as a function of the deposition time has been studied using height-height correlation function analysis from atomic force microscopy (AFM) images. The scaling exponents α, ß, 1/z and γ of the films were determined from AFM images. The local roughness exponent, α, was found to be ∼0.86 in the early stage of growth of Cu films deposited for 10 minutes while it increased to 0.95 with a longer time of deposition of 20 minutes and beyond this, it was nearly constant. Interface width w (rms roughness) scales with depositing time (t) as ∼ t ß , with the value of the growth exponent, ß, of 1.07 ± 0.11 and lateral correlation length ξ following ξ = t 1/z and the value of 1/z = 0.70 ± 0.10. These exponent values convey that the growth dynamics of PLD Cu films can be best described by a combination of local and non-local models under a shadowing mechanism and under highly sticking substrate conditions. From the scaling exponents and power spectral density function, it is concluded that the films follow a mound like growth mechanism which becomes prominent at longer deposition times. All the Cu films exhibited SPR properties where the SPR peak shifts towards red with increasing correlation length (ξ) whereas bandwidth increases initially with ξ and thereafter decreases gradually with ξ.

Metformin reverses early cortical network dysfunction and behavior changes in Huntington's disease.

Catching primal functional changes in early, 'very far from disease onset' (VFDO) stages of Huntington's disease is likely to be the key to a successful therapy. Focusing on VFDO stages, we assessed neuronal microcircuits in premanifest Hdh150 knock-in mice. Employing in vivo two-photon Ca2+ imaging, we revealed an early pattern of circuit dysregulation in the visual cortex - one of the first regions affected in premanifest Huntington's disease - characterized by an increase in activity, an enhanced synchronicity and hyperactive neurons. These findings are accompanied by aberrations in animal behavior. We furthermore show that the antidiabetic drug metformin diminishes aberrant Huntingtin protein load and fully restores both early network activity patterns and behavioral aberrations. This network-centered approach reveals a critical window of vulnerability far before clinical manifestation and establishes metformin as a promising candidate for a chronic therapy starting early in premanifest Huntington's disease pathogenesis long before the onset of clinical symptoms.

The role of Ca2+ in cell death caused by oxidative glutamate toxicity and ferroptosis.

Ca2+ ions play a fundamental role in cell death mediated by oxidative glutamate toxicity or oxytosis, a form of programmed cell death similar and possibly identical to other forms of cell death like ferroptosis. Ca2+ influx from the extracellular space occurs late in a cascade characterized by depletion of the intracellular antioxidant glutathione, increases in cytosolic reactive oxygen species and mitochondrial dysfunction. Here, we aim to compare oxidative glutamate toxicity with ferroptosis, address the signaling pathways that culminate in Ca2+ influx and cell death and discuss the proteins that mediate this. Recent evidence hints toward a role of the machinery responsible for store-operated Ca2+ entry (SOCE), which refills the endoplasmic reticulum (ER) after receptor-mediated ER Ca2+ release or other forms of store depletion. Pharmacological inhibition of SOCE or transcriptional downregulation of proteins involved in SOCE like the ER Ca2+ sensor STIM1, the plasma membrane Ca2+ channels Orai1 and TRPC1 and the linking protein Homer protects against oxidative glutamate toxicity and direct oxidative stress caused by hydrogen peroxide or 1-methyl-4-phenylpyridinium (MPP+) injury, a cellular model of Parkinson's disease. This suggests that SOCE inhibition might have some potential therapeutic effects in human disease associated with oxidative stress like neurodegenerative disorders.

The thiol switch C684 in Mitofusin-2 mediates redox-induced alterations of mitochondrial shape and respiration.

Mitofusin-2 (MFN2) is a GTPase in the outer mitochondrial membrane involved in the regulation of mitochondrial fusion and bioenergetics. MFN2 also plays a role in mitochondrial fusion induced by changes in the intracellular redox state. Adding oxidized glutathione (GSSG), the core cellular stress indicator, to mitochondrial preparations stimulates mitochondrial fusion by inducing disulphide bond-mediated oligomer formation of MFN2 and its homolog MFN1 which involve cysteine 684 (C684) of MFN2. Mitochondrial hyperfusion represents an adaptive stress response that confers transient protection by increasing mitochondrial ATP production but how this depends on the thiol switch C684 in MFN2 has not been investigated. We now studied mitochondrial function using high-resolution respirometry in cells stably expressing wildtype or C684A MFN2 in MFN2-deficient fibroblasts in response to alterations of the redox state. Empty vector and untransfected cells served as controls. A single treatment of cells with 100 µM hydrogen peroxide 24 h before analysis had no effect on wildtype cells, but normalized the otherwise increased respiration of knockout cells and significantly increased respiration in C684A cells. In line with this, treating permeabilized cells for 10 min with 1 mM GSH greatly reduced respiration only in C684A cells. Our data indicate that mutation of this cysteine which forms disulphide bridges in an oxidative state, apparently renders MFN2 more susceptible to alterations of the redox environment. It remains to be investigated whether other posttranslational modifications like glutathionylation might play an additional role.

Antioxidant responses in the earthworm Aporrectodea caliginosa of eastern Slovakia: application of principal component analysis as a tool to identify metal contaminated areas.

The soil of Slovak Republic is severely contaminated with heavy metals, creating hazards to soil health. In order to assess the current status with the prospect of selecting the appropriate treatment methods and land use, this investigation aimed to determine a panel of complementary and ecologically relevant biomarkers that reflect adverse biological responses towards terrestrial pollutants. To attain this objective, the concentration of reduced glutathione and enzymes of glutathione antioxidant system were assessed in clitellate earthworm, Aporrectodea caliginosa sampled from selected sites of eastern Slovakia along with the pH and total metal concentration (As, Cd, Pb, Cr, Hg, Mn, Fe, Co, Ni, Cu, Zn) of soils. Positive, significant (p < 0.05) induction of glutathione peroxidase, glutathione reductase, and glutathione-S-transferase activities and depletion of reduced glutathione level (negative correlation) were associated with the increased soil metal concentrations. Metal interference was found in the detoxification process and antioxidant defense mechanism does not efficiently counteract the oxidative stress induced by chronic metal exposure. The tested biomarkers confirmed sensitive and affective response to the pollution of soil contaminants, in this case metals. This has a potential use in ecotoxicological field monitoring. The proposed principal component analysis is a multivariate model of data analysis that represents a cost-effective approach to differentiate metalliferous soils of eastern Slovakia with different health status.

Fabrication of photoluminescent nc-Si:SiO2 thin films prepared by PLD.

In the present report, the structural, compositional, morphological, and photoluminescence properties of nanostructured non-stoichiometric silicon oxide (nc-Si:SiO2 or SiOx) thin films fabricated by pulsed-laser ablation of silicon in the presence of oxygen pressure, from 10-4 to 0.5 mbar, are presented. X-ray diffraction spectra and Raman spectra confirmed the formation of nanocrystalline Si within the films while electron diffraction X-ray spectroscopy confirmed the increase in oxygen content with increasing O2 pressure. Scanning electron microscopy images of the SiOx films showed spreading of the micron-sized clusters on the otherwise uniform background, while Raman maps confirm the presence of nanocrystalline Si in these clusters embedded in a uniform matrix comprising oxidized amorphous silicon. A systematic blue shift in the band gap energy from 1.55 to 2.80 eV was observed with increasing O2 pressure in the SiOx films due to a shift in the stoichiometry of the films from x = 0.03 to 2.14 respectively. The films with higher oxygen content exhibited broad and intense PL emissions with multiple peaks originating from quantum confined (QC) Si nanocrystals as well as oxygen defects like NBOH and VO centers. The variation in PL intensity as a function of excitation intensity displays an initial linear increase followed by saturation, a characteristic feature of emissions from QC nc-Si.

GluN3A promotes dendritic spine pruning and destabilization during postnatal development.

Synaptic rearrangements during critical periods of postnatal brain development rely on the correct formation, strengthening, and elimination of synapses and associated dendritic spines to form functional networks. The correct balance of these processes is thought to be regulated by synapse-specific changes in the subunit composition of NMDA-type glutamate receptors (NMDARs). Among these, the nonconventional NMDAR subunit GluN3A has been suggested to play a role as a molecular brake in synaptic maturation. We tested here this hypothesis using confocal time-lapse imaging in rat hippocampal organotypic slices and assessed the role of GluN3A-containing NMDARs on spine dynamics. We found that overexpressing GluN3A reduced spine density over time, increased spine elimination, and decreased spine stability. The effect of GluN3A overexpression could be further enhanced by using an endocytosis-deficient GluN3A mutant and reproduced by silencing the adaptor protein PACSIN1, which prevents the endocytosis of endogenous GluN3A. Conversely, silencing of GluN3A reduced spine elimination and favored spine stability. Moreover, reexpression of GluN3A in more mature tissue reinstated an increased spine pruning and a low spine stability. Mechanistically, the decreased stability in GluN3A overexpressing neurons could be linked to a failure of plasticity-inducing protocols to selectively stabilize spines and was dependent on the ability of GluN3A to bind the postsynaptic scaffold GIT1. Together, these data provide strong evidence that GluN3A prevents the activity-dependent stabilization of synapses thereby promoting spine pruning, and suggest that GluN3A expression operates as a molecular signal for controlling the extent and timing of synapse maturation.

Expression of cocaine-evoked synaptic plasticity by GluN3A-containing NMDA receptors.

Drug-evoked synaptic plasticity in the mesolimbic dopamine (DA) system reorganizes neural circuits that may lead to addictive behavior. The first cocaine exposure potentiates AMPAR excitatory postsynaptic currents (EPSCs) onto DA neurons of the VTA but reduces the amplitude of NMDAR-EPSCs. While plasticity of AMPAR transmission is expressed by insertion of calcium (Ca(2+))-permeable GluA2-lacking receptors, little is known about the expression mechanism for altered NMDAR transmission. Combining ex vivo patch-clamp recordings, mouse genetics, and subcellular Ca(2+) imaging, we observe that cocaine drives the insertion of NMDARs that are quasi-Ca(2+)-impermeable and contain GluN3A and GluN2B subunits. These GluN3A-containing NMDARs appear necessary for the expression of cocaine-evoked plasticity of AMPARs. We identify an mGluR1-dependent mechanism to remove these noncanonical NMDARs that requires Homer/Shank interaction and protein synthesis. Our data provide insight into the early cocaine-driven reorganization of glutamatergic transmission onto DA neurons and offer GluN3A-containing NMDARs as new targets in drug addiction.

Self-assembly of ketals of arjunolic acid into vesicles and fibers yielding gel-like dispersions.

Ten aliphatic and aromatic ketals of arjunolic acid, a renewable, nanosized triterpenic acid which is obtainable from Terminalia arjuna, have been synthesized upon condensation with aldehydes. Self-assembly properties of the ketals have been studied in a wide range of organic liquids. With the exception of the p-nitrobenzylidene derivative, low concentrations of the ketals self-assemble and form gel-like dispersions in many of the organic liquids examined. The morphologies of the assemblies, studied at different distance scales by optical, electron, and atomic-force microscopies, consisted of fibrillar networks and vesicles which were able to entrap 5(6)-carboxyfluorescein as a guest molecule. X-ray diffractograms indicate that the fibrillar objects are crystalline. A charge-transfer complex was formed from a 1:1 mixture of ketal derivatives with electron-donating and electron-accepting groups, and the 9-anthrylidene derivative in its fibrillar network dimerized upon irradiation. Results demonstrate that subtle changes in the ketal structures can lead to very different aggregation pathways.

Self-assembly of esters of arjunolic acid into fibrous networks and the properties of their organogels.

Nine esters of a naturally occurring triterpenoid, arjunolic acid (from Terminalia arjuna), with alkyl chains have been synthesized, and their self-assembly has been studied in organic liquids. All of the esters examined were found to be excellent gelators. No birefringence was detected in optical micrographs of the transparent toluene gels with 5% (w/w) ethyl arjunolate or 5% (w/w) p-nitrobenzyl arjunolate as the gelator, but a spherulitic-type pattern was seen for a gel of 1.2% (w/w) p-nitrobenzyl arjunolate in 1/1 (w/w) chloroform/cyclohexane. Electron microscope images revealed self-assembled fibrillar network (SAFIN) structures with right-handed helical ribbons in some gels. With increasing concentration of the gelators, the gel-to-sol transition temperature (T(gel)) increased and then approached plateau values. Differential scanning thermograms demonstrated that the heats for transition from transparent gels to sols of ethyl arjunolate or p-nitrobenzyl ajunolate in toluene are very small. Powder X-ray diffractograms revealed that the molecular packing in the SAFIN of the 5% (w/w) ethyl aijunoate in the toluene gel was amorphous and similar to the diffractogram recorded for the neat gelator. Although the diffractogram of neat p-nitrobenzyl arjunolate consisted of broad peaks, suggesting disordered packing, the low-angle peaks of the corresponding toluene gel were much sharper; these results indicate more crystalline packing in the SAFIN than in the neat gelator. The kinetics and growth of the transformation of sols of p-nitrobenzyl arjunolate in 1/1 (w/w) chloroform/cyclohexane to their gels have been investigated at different incubation temperatures by circular dichroism spectroscopy. The data have been analyzed to probe the mechanism of SAFIN formation and the relationship between the molecular structures of the esters of arjunolic acid and their abilities to function as gelators of a wide variety of organic liquids.