Disease evolution and risk-based disease trajectories in congestive heart failure patients.

Congestive Heart Failure (CHF) is among the most prevalent chronic diseases worldwide, and is commonly associated with comorbidities and complex health conditions. Consequently, CHF patients are typically hospitalized frequently, and are at a high risk of premature death. Early detection of an envisaged patient disease trajectory is crucial for precision medicine. However, despite the abundance of patient-level data, cardiologists currently struggle to identify disease trajectories and track the evolution patterns of the disease over time, especially in small groups of patients with specific disease subtypes. The present study proposed a five-step method that allows clustering CHF patients, detecting cluster similarity, and identifying disease trajectories, and promises to overcome the existing difficulties. This work is based on a rich dataset of patients' records spanning ten years of hospital visits. The dataset contains all the health information documented in the hospital during each visit, including diagnoses, lab results, clinical data, and demographics. It utilizes an innovative Cluster Evolution Analysis (CEA) method to analyze the complex CHF population where each subject is potentially associated with numerous variables. We have defined sub-groups for mortality risk levels, which we used to characterize patients' disease evolution by refined data clustering in three points in time over ten years, and generating patients' migration patterns across periods. The results elicited 18, 23, and 25 clusters respective to the first, second, and third visits, uncovering clinically interesting small sub-groups of patients. In the following post-processing stage, we identified meaningful patterns. The analysis yielded fine-grained patient clusters divided into several finite risk levels, including several small-sized groups of high-risk patients. Significantly, the analysis also yielded longitudinal patterns where patients' risk levels changed over time. Four types of disease trajectories were identified: decline, preserved state, improvement, and mixed-progress. This stage is a unique contribution of the work. The resulting fine partitioning and longitudinal insights promise to significantly assist cardiologists in tailoring personalized interventions to improve care quality. Cardiologists could utilize these results to glean previously undetected relationships between symptoms and disease evolution that would allow a more informed clinical decision-making and effective interventions.

Immobilized random peptide mixtures exhibit broad antimicrobial activity with high selectivity.

In the current study, we evaluated the antimicrobial activity of randomly-sequenced peptide mixtures (RPMs) bearing hydrophobic and cationic residues that were immobilized on beads. We showed that these beads exhibit high and broad bactericidal activity against various pathogenic bacteria while possessing minimal hemolytic activity.

Combining micelle-clay sorption to solar photo-Fenton processes for domestic wastewater treatment.

A tertiary treatment of effluent from a biological domestic wastewater treatment plant was tested by combining filtration and solar photocatalysis. Adsorption was carried out by a sequence of two column filters, the first one filled with granular activated carbon (GAC) and the second one with granulated nano-composite of micelle-montmorillonite mixed with sand (20:100, w/w). The applied solar advanced oxidation process was homogeneous photo-Fenton photocatalysis using peroxymonosulfate (PMS) as oxidant agent. This combination of simple, robust, and low-cost technologies aimed to ensure water disinfection and emerging contaminants (ECs, mainly pharmaceuticals) removal. The filtration step showed good performances in removing dissolved organic matter and practically removing all bacteria such as Escherichia coli and Enterococcus faecalis from the secondary treated water. Solar advanced oxidation processes were efficient in elimination of trace levels of ECs. The final effluent presented an improved sanitary level with acceptable chemical and biological characteristics for irrigation.

Removal of cyanobacteria and cyanotoxins from lake water by composites of bentonite with micelles of the cation octadecyltrimethyl ammonium (ODTMA).

Cyanobacteria and their toxins present potential hazard to consumers of water from lakes, reservoirs and rivers, thus their removal via water treatment is essential. The capacity of nano-composites of Octadecyltrimethyl-ammonium (ODTMA) complexed with clay to remove cyanobacterial and their toxins from laboratory cultures and from lake water, was evaluated. Column filters packed with micelles of ODTMA complexed with bentonite and granulated were shown to significantly reduce the number of cyanobacteria cells or filaments and their corresponding toxins from laboratory cultures. Fluorescence measurements demonstrated that cyanobacteria cells lost their metabolic activity (photosynthesis) upon exposure to the micelle (ODTMA)-bentonite complex, or ODTMA monomers. The complex efficiently removed cyanobacteria toxins with an exceptional high removal rate of microcystins. The effectiveness of the complex in elimination of cyanobacteria was further demonstrated with lake water containing cyanobacteria and other phytoplankton species. These results and model calculations suggest that filters packed with granulated composites can secure the safety of drinking water in case of a temporary bloom event of toxic cyanobacteria.

Combined adsorption and degradation of the off-flavor compound 2-methylisoborneol in sludge derived from a recirculating aquaculture system.

Off-flavor in fish poses a serious threat for the aquaculture industry. In the present study, removal of 2-methylisoborneol (MIB), an off-flavor causing compound, was found to be mediated by adsorption and bacterial degradation in sludge derived from an aquaculture system. A numerical model was developed which augmented Langmuir equations of kinetics of adsorption/desorption of MIB with first order degradation kinetics. When laboratory-scale reactors, containing sludge from the aquaculture system, were operated in a recirculating mode, MIB in solution was depleted to undetectable levels within 6 days in reactors with untreated sludge, while its depletion was incomplete in reactors with sterilized sludge. When operated in an open flow mode, removal of MIB was significantly faster in reactors with untreated sludge. Efficient MIB removal was evident under various conditions, including ambient MIB levels, flow velocities and sludge loads. When operated in an open flow mode, the model successfully predicted steady MIB removal rates with time. During steady state conditions, most of the MIB removal was found to be due to microbial degradation of the adsorbed MIB. Findings obtained in this study can be used in the design of reactors for removal of off-flavor compounds from recirculating aquaculture systems.

Paracetamol biodegradation by activated sludge and photocatalysis and its removal by a micelle-clay complex, activated charcoal, and reverse osmosis membranes.

Kinetic studies on the stability of the pain killer paracetamol in Al-Quds activated sludge demonstrated that paracetamol underwent biodegradation within less than one month to furnish p-aminophenol in high yields. Characterizations of bacteria contained in Al-Quds sludge were accomplished. It was found that Pseudomonas aeruginosa is the bacterium most responsible for the biodegradation of paracetamol to p-aminophenol and hydroquinone. Batch adsorptions of paracetamol and its biodegradation product (p-aminophenol) by activated charcoal and a composite micelle (octadecyltrimethylammonium)-clay (montmorillonite) were determined at 25°C. Adsorption was adequately described by a Langmuir isotherm, and indicated better efficiency of removal by the micelle-clay complex. The ability of bench top reverse osmosis (RO) plant as well as advanced membrane pilot plant to remove paracetamol was also studied at different water matrixes to test the effect of organic matter composition. The results showed that at least 90% rejection was obtained by both plants. In addition, removal of paracetamol from RO brine was investigated by using photocatalytic processes; optimal conditions were found to be acidic or basic pH, in which paracetamol degraded in less than 5 min. Toxicity studies indicated that the effluent and brine were not toxic except for using extra low energy membrane which displayed a half maximal inhibitory concentration (IC-50) value of 80%.

Stability and removal of spironolactone from wastewater.

Stability and removal of spironolactone (SP) from wastewater produced at Al-Quds University Campus were investigated. Kinetic studies on both pure water and wastewater coming from secondary treatment (activated sludge) demonstrated that the potassium-sparing diuretic (water pill), spironolactone, underwent degradation to its hydrolytic derivative, canrenone, in both media. The first-order hydrolysis rate of SP in activated sludge at 25°C (3.80 × 10(-5) s(-1)) was about 49-fold larger than in pure water (7.4 × 10(-7) s(-1)). The overall performance of the wastewater treatment plant (WWTP) installed in the University Campus was assessed showing that more than 90% of spiked SP was removed together with its newly identified metabolites. In order to look for a technology to supplement or replace ultra-filtration membranes, the effectiveness of adsorption and filtration by micelle-clay filters for removing SP was tested in comparison with activated charcoal. Batch adsorption in aqueous suspensions was well described by Langmuir isotherms, showing a better removal by the micelle-clay material. Filtration of SP water solutions by columns filled with a mixture of sand and a micelle-clay complex showed complete removal of the drug at concentrations higher than in sand/activated-charcoal filled filters.

Stability and removal of atorvastatin, rosuvastatin and simvastatin from wastewater.

Atorvastatin (ATO), rosuvastatin (RST) and simvastatin (SIM) are commonly used drugs that belong to the statin family (lowering human blood cholesterol levels) and have been detected as contaminants in natural waters. Stability and removal of ATO, RST and SIM from spiked wastewater produced at the Al-Quds University campus were investigated. All three statins were found to undergo degradation in wastewater (activated sludge). The degradation reactions of the three drugs in wastewater at room temperature follow first-order kinetics with rate constants of 2.2 × 10⁻7 s⁻¹ (ATO), 1.8 × 10⁻7 s⁻¹ (RST) and 1.8 × 10⁻6 s⁻¹ (SIM), which are larger than those obtained in pure water under the same conditions, 1.9 × 10⁻8 s⁻¹ (ATO), 2.2 × 10⁻8 s⁻¹ (RST) and 6.2 × 10⁻7 s⁻¹ (SIM). Degradation products were identified by LC-MS and LC/MS/MS. The overall performance of the wastewater treatment plant (WWTP) installed in the Al-Quds University campus towards the removal of these drugs was assessed showing that more than 90% of spiked ATO, RST and SIM were removed. In order to evaluate the efficiency of alternative removal methods to replace ultra-filtration membranes, adsorption isotherms for the three statins were investigated using both activated carbon and clay-micelle complex as adsorbents. The batch adsorption isotherms for the three statins were found to fit the Langmuir equation, with a larger number of adsorption sites and binding affinity for micelle-clay composite compared with activated carbon and filtration experiments of the three statins and their corresponding metabolites demonstrated a more efficient removal by micelle-clay filters.

Removal of amoxicillin and cefuroxime axetil by advanced membranes technology, activated carbon and micelle-clay complex.

Two antibacterials, amoxicillin trihydrate and cefuroxime axetil spiked into wastewater were completely removed by sequential wastewater treatment plant's membranes, which included activated sludge, ultrafiltration (hollow fibre and spiral wound membranes with 100 and 20 kDa cut-offs), activated carbon column and reverse osmosis. Adsorption isotherms in synthetic water which employed activated carbon and micelle-clay complex (octadecyltrimethylammonium-montmorillonite) as adsorbents fitted the Langmuir equation. Qmax of 100 and 90.9 mg g(-1), and K values of 0.158 and 0.229 L mg(-1) were obtained for amoxicillin trihydrate using activated carbon and micelle-clay complex, respectively. Filtration of antibacterials in the ppm range, which yielded variable degrees of removal depending on the volumes passed and flow rates, was simulated and capacities for the ppb range were estimated. Stability study in pure water and wastewater revealed that amoxicillin was totally stable for one month when kept at 37°C, whereas cefuroxime axetil underwent slow hydrolysis to cefuroxime.

Filtration of triazine herbicides by polymer-clay sorbents: coupling an experimental mechanistic approach with empirical modeling.

Triazine herbicides detected in surface and groundwater pose environmental and health risks. Removal of triazine herbicides (simazine, atrazine and terbuthylazine) by polymer-clay composites was studied and modeled. Their binding by a poly 4-vinyl pyridine co styrene-montmorillonite (HPVP-CoS-MMT) composite was especially high due to specific interactions between the herbicides and polymer, mainly hydrogen bonds and π-π stacking. The binding kinetics to the composite was in the order of simazine > atrazine > terbuthylazine, which was in accord with their equilibrium Langmuir binding coefficients; 44,000, 17,500 and 16,500 M(-1), respectively, which correlated with herbicide accessibility to form specific interaction with the polymer. Simazine binding kinetics to the composite was significantly faster than to granulated activated carbon (GAC), reaching 93% vs 38% of the maximal adsorption within 10 min, respectively. Herbicide filtration by composite columns was adequately fitted by a model which considers convection and employs Langmuir formalism for kinetics of adsorption/desorption. Filtration of simazine (10 µg L(-1)) by composite columns (40 cm long, which included 26 g composite mixed with sand 1:40 (weight ratio)), was well predicted by the model with nearly 120 L purified, i.e., effluent concentrations were below regulation limit (3 µg L(-1)). Effluent concentrations from GAC columns exceeded the limit after filtering 5 L. Experimental results and model predictions suggest that while GAC has a high capacity for simazine binding, the composite has higher affinity towards the herbicide and its adsorption is faster, which yields more efficient filtration by composite columns.

Removal of waterborne microorganisms by filtration using clay-polymer complexes.

Clay-polymer composites were designed for use in filtration processes for disinfection during the course of water purification. The composites were formed by sorption of polymers based on starch modified with quaternary ammonium ethers onto the negatively charged clay mineral bentonite. The performance of the clay-polymer complexes in removal of bacteria was strongly dependent on the conformation adopted by the polycation on the clay surface, the charge density of the polycation itself and the ratio between the concentrations of clay and polymer used during the sorption process. The antimicrobial effect exerted by the clay-polymer system was due to the cationic monomers adsorbed on the clay surface, which resulted in a positive surface potential of the complexes and charge reversal. Clay-polymer complexes were more toxic to bacteria than the polymers alone. Filtration employing our optimal clay-polymer composite yielded 100% removal of bacteria after the passage of 3L, whereas an equivalent filter with granular activated carbon (GAC) hardly yielded removal of bacteria after 0.5L. Regeneration of clay-polymer complexes saturated with bacteria was demonstrated. Modeling of the filtration processes permitted to optimize the design of filters and estimation of experimental conditions for purifying large water volumes in short periods.

Stability and removal of dexamethasone sodium phosphate from wastewater using modified clays.

Stability and removal of dexamethasone sodium phosphate (DSP) from wastewater produced at Al-Quds University Campus were investigated. Kinetic studies in both pure water and wastewater coming from secondary treatment (activated sludge) demonstrated that the anti-inflammatory DSP underwent degradation to its hydrolytic derivative, dexamethasone, in both media. The first-order hydrolysis rate of DSP in activated sludge at 25 degrees C (3.80 x 10(-6) s-1) was about 12-fold larger than in pure water (3.25 x 10(-7) s-1). The overall performance of the wastewater treatment plant (WWTP) installed in the University Campus was also assessed showing that 90% of spiked DSP was removed together with its newly identified metabolites by the ultra-filtration (UF) system, which consists of a UF hollow fibre (HF) with a 100-kDa cutoff membrane as the pre-polishing stage for the UF spiral wound with a 20-kDa cutoffmembrane. In testing other technologies, the effectiveness of adsorption and filtration by micelle-clay (MC) preparation for removing DSP was ascertained in comparison with activated charcoal. Batch adsorption in aqueous suspensions of the MC composite and activated carbon was well described by Langmuir isotherms showing the best results for MC material. Filtration of DSP water solutions demonstrated a significant advantage of columns filled in with a mixture of sand and MC complex in comparison with activated carbon/sand filters.

Removal of Cr(VI) from aqueous environments using micelle-clay adsorption.

Removal of Cr(VI) from aqueous solutions under different conditions was investigated using either clay (montmorillonite) or micelle-clay complex, the last obtained by adsorbing critical micelle concentration of octadecyltrimethylammonium ions onto montmorillonite. Batch experiments showed the effects of contact time, adsorbent dosage, and pH on the removal efficiency of Cr(VI) from aqueous solutions. Langmuir adsorption isotherm fitted the experimental data giving significant results. Filtration experiments using columns filled with micelle-clay complex mixed with sand were performed to assess Cr(VI) removal efficiency under continuous flow at different pH values. The micelle-clay complex used in this study was capable of removing Cr(VI) from aqueous solutions without any prior acidification of the sample. Results demonstrated that the removal effectiveness reached nearly 100% when using optimal conditions for both batch and continuous flow techniques.

Efficiency of membrane technology, activated charcoal, and a micelle-clay complex for removal of the acidic pharmaceutical mefenamic acid.

The efficiency of sequential advanced membrane technology wastewater treatment plant towards removal of a widely used non-steroid anti-inflammatory drug (NSAID) mefenamic acid was investigated. The sequential system included activated sludge, ultrafiltration by hollow fibre membranes with 100 kDa cutoff, and spiral wound membranes with 20 kDa cutoff, activated carbon and a reverse osmosis (RO) unit. The performance of the integrated plant showed complete removal of mefenamic acid from spiked wastewater samples. The activated carbon column was the most effective component in removing mefenamic acid with a removal efficiency of 97.2%. Stability study of mefenamic acid in pure water and Al-Quds activated sludge revealed that the anti-inflammatory drug was resistant to degradation in both environments. Batch adsorption of mefenamic acid by activated charcoal and a composite micelle (otadecyltrimethylammonium (ODTMA)-clay (montmorillonite) was determined at 25.0°C. Langmuir isotherm was found to fit the data with Qmax of 90.9 mg g(-1) and 100.0 mg g(-1) for activated carbon and micelle-clay complex, respectively. Filtration experiment by micelle-clay columns mixed with sand in the mg L(-1) range revealed complete removal of the drug with much larger capacity than activated carbon column. The combined results demonstrated that an integration of a micelle-clay column in the plant system has a good potential to improve the removal efficiency of the plant towards NSAID drugs such as mefenamic acid.

Efficiency of advanced wastewater treatment plant system and laboratory-scale micelle-clay filtration for the removal of ibuprofen residues.

The efficiency of Al-Quds Waste Water Treatment Plant (WWTP), which includes sequential elements as activated sludge, ultrafiltration, activated carbon column and reverse osmosis, to remove spiked ibuprofen, a non steroid anti inflammatory drug (NSAID), was investigated. Kinetic studies in pure water and in the activated sludge indicated that the drug was stable during one month of observation. Besides, the overall performance of the integrated plant showed complete removal of ibuprofen from wastewater. Activated carbon column, which was the last element in the sequence before the reverse osmosis system, yielded 95.7% removal of ibuprofen. Batch adsorptions of the drug by using either activated charcoal or composite micelle-clay system were determined at 25°C and well described by Langmuir isotherms. Octadecyltrimethylammonium (ODTMA) bromide and montmorillonite were used to prepare the micelle-clay adsorbent, for which the adsorption kinetics are much faster than activated charcoal. Results suggest that integrating clay-micelle complex filters within the existing WWTP may be promising in improving removal efficiency of the NSAID.

Factors affecting the design of slow release formulations of herbicides based on clay-surfactant systems. A methodological approach.

A search for clay-surfactant based formulations with high percentage of the active ingredient, which can yield slow release of active molecules is described. The active ingredients were the herbicides metribuzin (MZ), mesotrione (MS) and flurtamone (FL), whose solubilities were examined in the presence of four commercial surfactants; (i) neutral: two berols (B048, B266) and an alkylpolyglucoside (AG6202); (ii) cationic: an ethoxylated amine (ET/15). Significant percent of active ingredient (a.i.) in the clay/surfactant/herbicide formulations could be achieved only when most of the surfactant was added as micelles. MZ and FL were well solubilized by berols, whereas MS by ET/15. Sorption of surfactants on the clay mineral sepiolite occurred mostly by sorption of micelles, and the loadings exceeded the CEC. Higher loadings were determined for B266 and ET/15. The sorption of surfactants was modeled by using the Langmuir-Scatchard equation which permitted the determination of binding coefficients that could be used for further predictions of the sorbed amounts of surfactants under a wide range of clay/surfactant ratios. A possibility was tested of designing clay-surfactant based formulations of certain herbicides by assuming the same ratio between herbicides and surfactants in the formulations as for herbicides incorporated in micelles in solution. Calculations indicated that satisfactory FL formulations could not be synthesized. The experimental fractions of herbicides in the formulations were in agreement with the predicted ones for MS and MZ. The validity of this approach was confirmed in in vitro release tests that showed a slowing down of the release of a.i. from the designed formulations relative to the technical products. Soil dissipation studies with MS formulations also showed improved bioactivity of the clay-surfactant formulation relative to the commercial one. This methodological approach can be extended to other clay-surfactant systems for encapsulation and slow release of target molecules of interest.

Removal of diclofenac potassium from wastewater using clay-micelle complex.

The presence of an ionized carboxyl group in the widely used non-steroidal anti-inflammatory (NSAID) drug diclofenac potassium results in a high mobility of diclofenac and in its low sorption under conditions of slow sand filtration or subsoil passage. No diclofenac degradation was detected in pure water or sludge during one month. Tertiary treatments of wastewater indicated that the effective removal of diclofenac was by reverse osmosis, but the removal by activated carbon was less satisfactory. This study presents an efficient method for the removal of diclofenac from water by micelle-clay composites that are positively charged, have a large surface area and include large hydrophobic domains. Adsorption of diclofenac in dispersion by charcoal and a composite micelle (otadecyltrimethylammonium [ODTMA] and clay [montmorillonite]) was investigated. Analysis by the Langmuir isotherm revealed that charcoal had a somewhat larger number of adsorption sites than the composite, but the latter had a significantly larger binding affinity for diclofenac. Filtration experiments on a solution containing 300 ppm diclofenac demonstrated poor removal by activated carbon, in contrast to very efficient removal by micelle-clay filters. In the latter case the weight of removed diclofenac exceeded half that of ODTMA in the filter. Filtration of diclofenac solutions at concentrations of 8 and 80 ppb yielded almost complete removal at flow rates of 30 and 60 mL min(-1). One kilogram of ODTMA in the micelle-clay filter has been estimated to remove more than 99% of diclofenac from a solution of 100 ppb during passage of more than 100 m3.

Membrane-proximal external HIV-1 gp41 motif adapted for destabilizing the highly rigid viral envelope.

Electron microscopy structural determinations suggest that the membrane-proximal external region (MPER) of glycoprotein 41 (gp41) may associate with the HIV-1 membrane interface. It is further proposed that MPER-induced disruption and/or deformation of the lipid bilayer ensue during viral fusion. However, it is predicted that the cholesterol content of this membrane (∼45 mol %) will act against MPER binding and restructuring activity, in agreement with alternative structural models proposing that the MPER constitutes a gp41 ectodomain component that does not insert into the viral membrane. Here, using MPER-based peptides, we test the hypothesis that cholesterol impedes the membrane association and destabilizing activities of this gp41 domain. To that end, partitioning and leakage assays carried out in lipid vesicles were combined with x-ray reflectivity and grazing-incidence diffraction studies of monolayers. CpreTM, a peptide combining the carboxyterminal MPER sequence with aminoterminal residues of the transmembrane domain, bound and destabilized effectively cholesterol-enriched membranes. Accordingly, virion incubation with this peptide inhibited cell infection potently but nonspecifically. Thus, CpreTM seems to mimic the envelope-perturbing function of the MPER domain and displays antiviral activity. As such, we infer that CpreTM bound to cholesterol-enriched membranes would represent a relevant target for anti-HIV-1 immunogen and inhibitor development.

Destabilization exerted by peptides derived from the membrane-proximal external region of HIV-1 gp41 in lipid vesicles supporting fluid phase coexistence.

The human immunodeficiency virus (HIV) envelope is enriched in cholesterol and sphingomyelin, two lipids that sustain the formation of laterally segregated liquid-ordered fluid domains in model systems. Several evidences indicate that the high lipid order existing at the envelope may play a role in HIV pathogenesis. A putative mechanism might involve the modulation of the membrane-perturbing function of the gp41 membrane-proximal external region (MPER). To test such hypothesis, we investigate here the effect of lipid phase coexistence on the membrane-restructuring properties of NpreTM and CpreTM, two peptides based on the amino- and carboxy-terminal MPER sequences, respectively. Fluid phase coexistence elicited the fusogenic activity of NpreTM at high membrane doses and stimulated "graded" leakage at low doses. By comparison, the effect on CpreTM was restricted to an enhancement of "all-or-none" leakage that was consistent with the promotion of its surface aggregation. Confocal microscopy of single vesicles revealed the preference of both peptides for liquid-disordered domains. Accordingly, we speculate that confinement into envelope fluid nanodomains might boost the distinct capacities of HIV MPER hydrophobic modules for inducing membrane defects during fusion.

Distinct mechanisms of lipid bilayer perturbation induced by peptides derived from the membrane-proximal external region of HIV-1 gp41.

The conserved, membrane-proximal external region (MPER) of the human immunodeficiency virus type-1 envelope glycoprotein 41 subunit is required for fusogenic activity. It has been proposed that MPER functions by disrupting the virion membrane. Supporting its critical role in viral entry as a membrane-bound entity, MPER constitutes the target for broadly neutralizing antibodies that have evolved mechanisms to recognize membrane-inserted epitopes. We have analyzed here the molecular mechanisms of membrane permeabilization induced by N-preTM and PreTM-C, two peptides derived from MPER sequences showing a tendency to associate with the bilayer interface or to transfer into the hydrocarbon core, respectively. Both peptides contained the full epitope sequence recognized by the 4E10 monoclonal antibody (MAb4E10), which was subsequently used to probe peptide accessibility from the water phase. Capacities of N-preTM and PreTM-C for associating with vesicles and inducing their permeabilization were comparable. However, MAb4E10 specifically blocked the permeabilization induced by N-preTM but did not appreciably affect that induced by PreTM-C. Supporting the existence of different membrane-bound lytic structures, N-preTM was running as a monomer on SDS-PAGE and induced the graded release of vesicular contents, whereas PreTM-C migrated on SDS-PAGE as dimers and permeabilized vesicles following an all-or-none mechanism, reminiscent of that underlying melittin-induced membrane lysis. These results support the functional segmentation of gp41 membrane regions into hydrophobic subdomains, which might expose neutralizing epitopes and induce membrane-disrupting effects following distinct patterns during the fusion cascade.