The phenotypic landscape of essential human genes.

Understanding the basis for cellular growth, proliferation, and function requires determining the roles of essential genes in diverse cellular processes, including visualizing their contributions to cellular organization and morphology. Here, we combined pooled CRISPR-Cas9-based functional screening of 5,072 fitness-conferring genes in human HeLa cells with microscopy-based imaging of DNA, the DNA damage response, actin, and microtubules. Analysis of >31 million individual cells identified measurable phenotypes for >90% of gene knockouts, implicating gene targets in specific cellular processes. Clustering of phenotypic similarities based on hundreds of quantitative parameters further revealed co-functional genes across diverse cellular activities, providing predictions for gene functions and associations. By conducting pooled live-cell screening of ∼450,000 cell division events for 239 genes, we additionally identified diverse genes with functional contributions to chromosome segregation. Our work establishes a resource detailing the consequences of disrupting core cellular processes that represents the functional landscape of essential human genes.

Optical Pooled Screens in Human Cells.

Genetic screens are critical for the systematic identification of genes underlying cellular phenotypes. Pooling gene perturbations greatly improves scalability but is not compatible with imaging of complex and dynamic cellular phenotypes. Here, we introduce a pooled approach for optical genetic screens in mammalian cells. We use targeted in situ sequencing to demultiplex a library of genetic perturbations following image-based phenotyping. We screened a set of 952 genes across millions of cells for involvement in nuclear factor κB (NF-κB) signaling by imaging the translocation of RelA (p65) to the nucleus. Screening at a single time point across 3 cell lines recovered 15 known pathway components, while repeating the screen with live-cell imaging revealed a role for Mediator complex subunits in regulating the duration of p65 nuclear retention. These results establish a highly multiplexed approach to image-based screens of spatially and temporally defined phenotypes with pooled libraries.

Exploring the fluorescence properties of tellurium-containing molecules and their advanced applications.

This review article explores the fascinating realm of fluorescence using organochalcogen molecules, with a particular emphasis on tellurium (Te). The discussion encompasses the underlying mechanisms, structural motifs influencing fluorescence, and the applications of these intriguing phenomena. This review not only elucidates the current state of knowledge but also identifies avenues for future research, thereby serving as a valuable resource for researchers and enthusiasts in the field of fluorescence chemistry with a focus on Te-based molecules. By highlighting challenges and prospects, this review sparks a conversation on the transformative potential of Te-containing compounds across different fields, ranging from environmental solutions to healthcare and materials science applications. This review aims to provide a comprehensive understanding of the distinct fluorescence behaviors exhibited by Te-containing compounds, contributing valuable insights to the evolving landscape of chalcogen-based fluorescence research.

Pooled genetic screens with image-based profiling.

Spatial structure in biology, spanning molecular, organellular, cellular, tissue, and organismal scales, is encoded through a combination of genetic and epigenetic factors in individual cells. Microscopy remains the most direct approach to exploring the intricate spatial complexity defining biological systems and the structured dynamic responses of these systems to perturbations. Genetic screens with deep single-cell profiling via image features or gene expression programs have the capacity to show how biological systems work in detail by cataloging many cellular phenotypes with one experimental assay. Microscopy-based cellular profiling provides information complementary to next-generation sequencing (NGS) profiling and has only recently become compatible with large-scale genetic screens. Optical screening now offers the scale needed for systematic characterization and is poised for further scale-up. We discuss how these methodologies, together with emerging technologies for genetic perturbation and microscopy-based multiplexed molecular phenotyping, are powering new approaches to reveal genotype-phenotype relationships.

Chitooligosaccharide-catechin conjugate loaded liposome using different stabilising agents: characteristics, stability, and bioactivities.

AIM: To determine the optimum condition for preparing chitooligosaccharide-catechin conjugate (COS-CAT) liposomes using different stabilising agents. METHODS: COS-CAT liposomes (0.1-1%, w/v) were prepared using soy phosphatidylcholine (SPC) (50-200 mM) and glycerol or cholesterol (25-100 mg). Encapsulation efficiency (EE), loading capacity (LC), physicochemical characteristics, FTIR spectra, thermal stability, and structure of COS-CAT liposomes were assessed. RESULTS: COS-CAT loaded liposome stabilised by cholesterol (COS-CAT-CHO) showed higher stability as shown by the highest EE (76.81%) and LC (4.57%) and the lowest zeta potential (ZP) (-76.51 mV), polydispersity index (PDI) (0.2674) and releasing efficiency (RE) (53.54%) (p < 0.05). COS-CAT-CHO showed the highest retention and relative remaining bioactivities of COS-CAT under various conditions (p < 0.05). FTIR spectra revealed the interaction between the choline group of SPC and -OH groups of COS-CAT. Phase transition temperature of COS-CAT-CHO was shifted to 184 °C, which was higher than others (p < 0.05). CONCLUSION: SPC and cholesterol-based liposome could be used as a promising vesicle for maintaining bioactivities of COS-CAT.

Inhibition Mechanism of Chitooligosaccharide-Polyphenol Conjugates toward Polyphenoloxidase from Shrimp Cephalothorax.

Crustaceans are perishable with a short shelf-life. They are prone to deterioration after capture, particularly during handling, processing, and storage due to melanosis caused by polyphenoloxidase (PPO). Therefore, inhibitory effects of chitooligosaccharide (CHOS) in comparison with CHOS-catechin (CHOS-CAT), CHOS-epigallocatechin gallate (CHOS-EGCG), and CHOS-gallic acid (CHOS-GAL) conjugates on Pacific white shrimp cephalothorax PPO were studied. IC50 of CHOS-CAT (0.32 mg/mL) toward PPO was less than those of all conjugates tested (p < 0.05). CHOS-CAT exhibited the mixed-type inhibition. Kic (0.58 mg/mL) and Kiu (0.02 mg/mL) of CHOS-CAT were lower than those of other conjugates (p < 0.05). CHOS-CAT showed static fluorescence-quenching, suggesting a change in micro-environment around the active site of PPO. Moreover, CHOS-CAT was linked with various amino acid residues, including Tyr208 or Tyr209 of proPPO via van der Waals, hydrophobic interaction, and hydrogen bonding as elucidated by the molecular docking of proPPO. Although CHOS-CAT had the highest PPO inhibitory activity, it showed a lower binding energy (-8.5 kcal/mol) than other samples, except for CHOS-EGCG (-10.2 kcal/mol). Therefore, CHOS-CAT could act as an anti-melanosis agent in shrimp and other crustaceans to prevent undesirable discoloration associated with quality losses.

Exploring haplotype block structure, runs of homozygosity, and effective population size among dairy cattle breeds of India.

The present study aimed to explore haplotype structure, runs of homozygosity (ROH), effective population size and persistence of gametic phase among three indigenous dairy cattle breeds, viz., Sahiwal (n = 19), Tharparkar (n = 17), and Gir (n = 16) by using BovineHD single nucleotide polymorphism (SNP) genotyping assay. The filtered SNPs after quality control ranged from 44% in Sahiwal to 53% in Gir. The highest number of haplotype blocks was observed in Tharparkar (15,640) and the lowest in Sahiwal (8027) spanning 17.3% and 7.8% of genome, respectively. The average block length was found close to 26 kb which suggests that multiple recombination events fragmented the ancestral haplotypes into smaller sizes. Gir cattle had the largest number of runs of homozygosity (ROH) regions (1762) followed by Tharparkar (1528) and Sahiwal (1138). Without pedigree information, inbreeding coefficients estimated from ROH (FROH) revealed that Gir had the highest FROH (0.099) proposing more inbreeding rate in this population. Effective population size (Ne) decreased slowly over the last 60 generations and at 13 generations ago; Ne was estimated as 70 for all the three dairy breeds. The highest gametic phase correlation (r = 0.78) was observed for Sahiwal and Tharparkar breed pair suggesting formulation of multi-breed reference population for successful implementation of genomic selection among dairy breeds. The decline in effective population size among native Indian cattle breeds may help in formulating strategies for conservation and genetic improvement of native germplasm for future use.

Undesirable discoloration in edible fish muscle: Impact of indigenous pigments, chemical reactions, processing, and its prevention.

Fish is rich in proteins and lipids, especially those containing polyunsaturated fatty acids, which made them vulnerable to chemical or microbial changes associated with quality loss. Meat color is one of vital criteria indicating the freshness, quality, and acceptability of the meat. Color of meat is governed by the presence of various pigments such as hemoglobin, myoglobin (Mb), and so on. Mb, particularly oxy-form, is responsible for the bright red color of fish muscle, especially tuna, and dark fleshed fish, while astaxanthin (AXT) directly determines the color of salmonids muscle. Microbial spoilage and chemical changes such as oxidation of lipid/proteins result in the autoxidation of Mb or fading of AXT, leading to undesirable color with lower acceptability. The discoloration has been affected by chemical composition, post-harvesting handling or storage, processing, cooking, and so on . To tackle discoloration of fish meat, vacuum or modified atmospheric packaging, low- or ultralow-temperature storage, uses of artificial and natural additives have been employed. This review article provides information regarding the factors affecting color and other quality aspects of fish muscle. Moreover, promising methodologies used to control discoloration are also focused.

Chitooligosaccharide prevents vascular endothelial cell apoptosis by attenuation of endoplasmic reticulum stress via suppression of oxidative stress through Nrf2-SOD1 up-regulation.

CONTEXT: Endoplasmic reticulum (ER) stress contributes to endothelium pathological conditions. Chitooligosaccharides (COS) have health benefits, but their effect on endothelial cells is unknown. We demonstrate for the first time a protective effect of COS against ER-induced endothelial cell damage. OBJECTIVE: To evaluate the protective effect of COS on ER stress-induced apoptosis in endothelial cells. MATERIAL AND METHODS: Endothelial (EA.hy926) cells were pre-treated with COS (250 or 500 µg/mL) for 24 h, and then treated with 0.16 µg/mL of Tg for 24 h and compared to the untreated control. Apoptosis and necrosis were detected by Annexin V-FITC/propidium iodide co-staining. Reactive oxygen species (ROS) were measured with the DCFH2-DA and DHE probes. The protective pathway and ER stress markers were evaluated by reverse transcription-polymerase chain reaction, western blot, and immunofluorescence analyses. RESULTS: COS attenuated ER stress-induced cell death. The viability of EA.hy926 cells treated with Tg alone was 44.97 ± 1% but the COS pre-treatment increased cells viability to 74.74 ± 3.95% in the 250 µg/mL COS and 75.34 ± 2.4% in the 500 µg/mL COS treatments. Tg induced ER stress and ROS, which were associated with ER stress-mediated death. Interestingly, COS reduced ROS by upregulating nuclear factor-E2-related factor 2 (Nrf2), and the oxidative enzymes, superoxide dismutase1 (SOD1) and catalase. COS also suppressed up-regulation of the ER-related apoptosis protein, CHOP induced by Tg. CONCLUSIONS: COS protected against ER stress-induced apoptosis in endothelial cells by suppressing ROS and up-regulation Nrf2 and SOD1. These findings support the use of COS to protect endothelial cells.

Rapid quality deterioration of harpiosquillid mantis shrimp (Harpiosquilla raphidea) during iced storage.

Changes in physicochemical, textural, microbial, and sensory quality of harpiosquillid mantis shrimp (Harpiosquilla raphidea) (HMS) during 10 days of iced storage were studied. Weight and cooking losses were increased during storage (p < 0.05). Drastic decrease in myosin heavy chain was found after 2 days of storage. Increases in total volatile basic nitrogen, trimethylamine, peroxide value, and thiobarbituric acid reactive substances with coincidentally augmented pH were found during the storage (p < 0.05). For microbiological analyses, total viable counts exceeded the limit at day 6. Melanosis score increased with a decreased L* value as storage time increased. Rapid decreases in hardness, springiness, cohesiveness, gumminess, and chewiness were associated with pasty and softened texture, which was supported by looser arrangement of muscle fiber along with gapping. This was reconfirmed by lowered shear force. Based on the quality evaluation, HMS could maintain the freshness and quality for not longer than 2 days in ice.

Effect of sodium bicarbonate on textural properties and acceptability of gel from unwashed Asian sea bass mince.

Asian sea bass mince gels having different adjusted moisture/water content (80 and 85%; w/w) were prepared with addition of sodium bicarbonate (SB) at various concentrations (0, 0.05 and 0.1%; w/w). Fish mince gels of 80% water content added with 0.05 and 0.1% SB (G80-0.05 and G80-0.1, respectively) had the highest increase (135-139%) in breaking force (BrF) than the respective control gel (G80) (P < 0.05). For gel with 85% water content, a lower increase (17-28%) in BrF was found with the addition of SB as compared to their corresponding control (G85). Whiteness of all samples was continuously decreased with increasing amount of SB, however the water holding capacity was increased drastically with augmenting levels of SB, regardless of the water content (P < 0.05). A loss in the elasticity of gel was attained with the addition of SB as indicated by decreasing storage modulus. A finer and more compact network was detected in a gel containing SB, irrespective of water content. Based on sensory scores, gel having 85% water content added with 0.05 and 0.1% SB had similar acceptability to the control gel (G80) containing 80% water content (commercial level). Therefore, SB at the appropriate level could improve the gelling properties with higher water holding ability of the mince gel with high acceptability.

Vascular and immunopathological role of Asymmetric Dimethylarginine (ADMA) in Experimental Autoimmune Encephalomyelitis.

Asymmetric dimethylarginine (ADMA) is an endogenous nitric oxide synthase (NOS) inhibitor/uncoupler inducing vascular pathology. Vascular pathology is an important factor for the development and progression of CNS pathology of MS, yet the role of ADMA in MS remains elusive. Patients with multiple sclerosis (MS) are reported to have elevated blood levels of ADMA, and mice with experimental autoimmune encephalomyelitis (EAE, an animal model of MS) generated by auto-immunization of myelin oligodendrocyte glycoprotein (MOG) and blood-brain barrier (BBB) disruption by pertussis toxin also had increased blood ADMA levels in parallel with induction of clinical disease. To explore the role of ADMA in EAE pathogenesis, EAE mice were treated with a daily dose of ADMA. It is of special interest that ADMA treatment enhanced the BBB disruption in EAE mice and exacerbated the clinical and CNS disease of EAE. ADMA treatment also induced the BBB disruption and EAE disease in MOG-immunized mice even without pertussis toxin treatment, suggesting the role of ADMA in BBB dysfunction in EAE. T-cell polarization studies also documented that ADMA treatment promotes TH 1- and TH 17-mediated immune responses but without affecting Treg-mediated immune response in EAE mice as well as in in vitro T-cell culture. Taken together, these data, for the first time, document the vascular and immunopathogenic roles of ADMA in EAE, thus pointing to the potential of ADMA-mediated mechanism as a new target of potential therapy for MS.

Asymmetric dimethylarginine exacerbates cognitive dysfunction associated with cerebrovascular pathology.

Asymmetric dimethylarginine (ADMA), an endogenous inhibitor and uncoupler of nitric oxide synthase, has gained attention as a risk factor for cardiac disease, metabolic syndrome, and cerebrovascular disease. In this study, we investigated the role of systemic ADMA overburden in cerebromicrovascular pathology associated with cognitive dysfunction using APPSwDI transgenic mice expressing human ß-amyloid precursor protein Swedish (Tg-SwDI), a model of cerebrovascular ß-amyloidosis. To induce systemic overburden of ADMA, Tg-SwDI mice were treated with a daily dose of exogenous ADMA. ADMA treatment resulted in elevated ADMA levels in the blood and brain of Tg-SwDI mice. ADMA treatment induced the brain nitrosative stress and inflammation as well as enhanced the brain Aß deposition and cognitive impairment in Tg-SwDI mice. However, ADMA treatment had no such effects on wild type mice. ADMA treatment also exacerbated brain microvascular pathology in Tg-SwDI mice as observed by increased blood-brain barrier dysfunction, loss of tight junction proteins, increased endothelial stress fibers, and decreased microvessel density in the brain. In addition, similar observations were made in cultured human brain microvessel endothelial cells, where ADMA in the presence of VEGF-induced endothelial cell signaling for F-actin stress fiber inducing endothelial barrier dysfunction. Overall, these data document the potential role of ADMA in the cognitive pathology under conditions of cerebrovascular ß-amyloidosis.

Environmental pollution reducing strategy for scouring of undegummed sisal fibers using xylanase and pectinase enzymes.

This study was undertaken to investigate the potential of bioscouring in the processing of undegummed sisal fibers, using xylano-pectinolytic enzymes. Optimum bioscouring was obtained at pH 8.5 and 50 mM buffer molarity, using xylanase (10 IU) and pectinase (8 IU), with a material to liquor proportion of 1:25 (g:ml), EDTA (2 mM) and Tween 80 (0.5%), at 50 °C temperature with agitation rate of 55 rpm and treatment period of 60 min. Enzymatic treatment of sisal fibers enhanced the brightness and whiteness by 11.52 and 6.83%, respectively, and reduced the yellowness by 7.14% in comparison to control. The use of xylanase and pectinase enzymes completely replaced the chemical scouring method for removing non-cellulosic impurities. Thus, enzymatic scouring is energy saving and ecofriendly, since it completely eliminated the use of toxic chemicals used in alkaline scouring. An increase of 23.75% and 11.58% in brightness and whiteness of enzymatically scoured cum bleached fibers, as compared to chemically scoured cum bleached fibers was finally obtained, along with reduction in yellowness by 27.99%. This is the first report demonstrating environmentally sustainable enzymatic approach for scouring of undegummed sisal fibers, using enzymes, simultaneously produced from a bacterial isolate.

Preparation and characterisation of liposome loaded with chitosan-epigallocatechin gallate conjugate.

Liposomes (LS) were prepared using chitosan-epigallocatechin gallate (CE) conjugate (0.1 and 0.5%, w/v) and soy phosphatidylcholine (SPC)/cholesterol as a lipid phase (LP) (30 and 60 µmol mL-1). The encapsulation efficiency (EE), particle diameter, zeta potential, and polydispersity index of LS were observed. The highest EE (76.96%) was found when LS was prepared using 0.5% (w/v) of CE conjugate and 60 µmol mL-1 of LP (CELP-60-0.5) (p < 0.05). FTIR analysis showed the interaction between choline present in SPC and OH-groups of CE conjugate. The phase transition temperature of CELP-60-0.5 was 134.67 °C, higher than other samples (p < 0.05). CELP-60-0.5 showed inhibitory action against Gram-positive and Gram-negative bacteria. Higher retention of antioxidant and antimicrobial activities of CELP-60-0.5 was observed than unencapsulated CE conjugate sample when stored for 28 days at 30 °C (p < 0.05). LS might be used as an efficient vesicle for maintaining bioactivities of CE conjugate, plausibly when used as a preservative in foods.

Regulation of endothelial barrier integrity by redox-dependent nitric oxide signaling: Implication in traumatic and inflammatory brain injuries.

Nitric oxide (NO) synthesized by eNOS plays a key role in regulation of endothelial barrier integrity but underlying cell signaling pathway is not fully understood at present. Here, we report opposing roles of two different redox-dependent NO metabolites; peroxynitrite (ONOO-) vs. S-nitrosoglutathione (GSNO), in cell signaling pathways for endothelial barrier disruption. In cultured human brain microvessel endothelial cells (hBMVECs), thrombin induced F-actin stress fiber formation causes barrier disruption via activating eNOS. Thrombin induced eNOS activity participated in cell signaling (e.g. RhoA and calcium influx mediated phosphorylation of myosin light chain) for F-actin stress fiber formation by increasing ONOO- levels. On the other hand, thrombin had no effect on intracellular levels of S-nitrosoglutathione (GSNO), another cellular NO metabolite. However, exogenous GSNO treatment attenuated the thrombin-induced cell signaling pathways for endothelial barrier disruption, thus suggesting the role of a shift of NO metabolism (GSNO vs. ONOO-) toward ONOO- synthesis in cell signaling for endothelial barrier disruption. Consistent with these in vitro studies, in animal models of traumatic brain injury and experimental autoimmune encephalomyelitis (EAE), ONOO- scavenger treatment as well as GSNO treatment were effective for attenuation of BBB leakage, edema formation, and CNS infiltration of mononuclear cells. Taken together, these data document that eNOS-mediated NO production and following redox-dependent NO metabolites (ONOO- vs. GSNO) are potential therapeutic target for CNS microvascular disease (traumatic and inflammatory) pathologies.

Biological, nutritional, and therapeutic significance of Moringa oleifera Lam.

The genus Moringa Adans. comprises 13 species, of which Moringa oleifera Lam. native to India and cultivated across the world owing to its drought and frost resistance habit is widely used in traditional phytomedicine and as rich source of essential nutrients. Wide spectrum of phytochemical ingredients among leaf, flower, fruit, seed, seed oil, bark, and root depend on cultivar, season, and locality. The scientific studies provide insights on the use of M. oleifera with different aqueous, hydroalcoholic, alcoholic, and other organic solvent preparations of different parts for therapeutic activities, that is, antibiocidal, antitumor, antioxidant, anti-inflammatory, cardio-protective, hepato-protective, neuro-protective, tissue-protective, and other biological activities with a high degree of safety. A wide variety of alkaloid and sterol, polyphenols and phenolic acids, fatty acids, flavanoids and flavanol glycosides, glucosinolate and isothiocyanate, terpene, anthocyanins etc. are believed to be responsible for the pragmatic effects. Seeds are used with a view of low-cost biosorbent and coagulant agent for the removal of metals and microbial contamination from waste water. Thus, the present review explores the use of M. oleifera across disciplines for its prominent bioactive ingredients, nutraceutical, therapeutic uses and deals with agricultural, veterinarian, biosorbent, coagulation, biodiesel, and other industrial properties of this "Miracle Tree."

S-Nitrosoglutathione Mimics the Beneficial Activity of Endothelial Nitric Oxide Synthase-Derived Nitric Oxide in a Mouse Model of Stroke.

BACKGROUND: The nitric oxide (NO)-producing activity of endothelial nitric oxide synthase (eNOS) plays a significant role in maintaining endothelial function and protecting against the stroke injury. However, the activity of the eNOS enzyme and the metabolism of major NO metabolite S-nitrosoglutathione (GSNO) are dysregulated after stroke, causing endothelial dysfunction. We investigated whether an administration of exogenous of GSNO or enhancing the level of endogenous GSNO protects against neurovascular injury in wild-type (WT) and eNOS-null (endothelial dysfunction) mouse models of cerebral ischemia-reperfusion (IR). METHODS: Transient cerebral ischemic injury was induced by middle cerebral artery occlusion (MCAO) for 60 minutes in male adult WT and eNOS null mice. GSNO (0.1 mg/kg body weight, intravenously) or N6022 (GSNO reductase inhibitor, 5.0 mg/kg body weight, intravenously) was administered 30 minutes before MCAO in preinjury and at the reperfusion in postinjury studies. Brain infarctions, edema, and neurobehavioral functions were evaluated at 24 hours after the reperfusion. RESULTS: eNOS-null mice had a higher degree (P< .05) of injury than WT. Pre- or postinjury treatment with either GSNO or N6022 significantly reduced infarct volume, improved neurological and sensorimotor function in both WT and eNOS-null mice. CONCLUSION: Reduced brain infarctions and edema, and improved neurobehavioral functions by pre- or postinjury GSNO treatment of eNOS knock out mice indicate that GSNO can attenuate IR injury, likely by mimicking the eNOS-derived NO-dependent anti-ischemic and anti-inflammatory functions. Neurovascular protection by GSNO/N6022 in both pre- and postischemic injury groups support GSNO as a promising drug candidate for the prevention and treatment of stroke injury.

Use of ultrasonicated squid ovary powder as a replacer of egg white powder in cake.

Ultrasonicated squid ovary powder (USOP) was used to replace egg white powder (EWP) at different substitution levels (12.5-100%) and its effects on properties of batter and cake were investigated. High elastic modulus (G') and average bubble size of batter added with 100% USOP resulted in higher volume and lower baking loss, when compared to the control cake (100% EWP). For textural analysis, the lowest values of hardness, gumminess and chewiness were noticeable for cake containing 100% USOP. Crust showed the lower moisture content than crumb and bottom part of all the cakes. No difference in moisture content was observed for all parts of the cakes containing 25-100% USOP, while lower moisture content was obtained for cake added with 12.5% USOP and the control. The color difference (ΔE*) between the control and cake added with USOP was increased with increasing USOP levels. When 100% of USOP was added into cake, higher likeness score was obtained for firmness and overall likeness. Microstructure study of cake added with 100% USOP revealed that oil phase was distributed in gluten matrix more uniformly, when compared to the control. Thus, replacement of EWP with 100% USOP resulted in the production of cake with superior quality and increased overall acceptance by consumers.

Combination therapy of lovastatin and AMP-activated protein kinase activator improves mitochondrial and peroxisomal functions and clinical disease in experimental autoimmune encephalomyelitis model.

Recent studies report that loss and dysfunction of mitochondria and peroxisomes contribute to the myelin and axonal damage in multiple sclerosis (MS). In this study, we investigated the efficacy of a combination of lovastatin and AMP-activated protein kinase (AMPK) activator (AICAR) on the loss and dysfunction of mitochondria and peroxisomes and myelin and axonal damage in spinal cords, relative to the clinical disease symptoms, using a mouse model of experimental autoimmune encephalomyelitis (EAE, a model for MS). We observed that lovastatin and AICAR treatments individually provided partial protection of mitochondria/peroxisomes and myelin/axons, and therefore partial attenuation of clinical disease in EAE mice. However, treatment of EAE mice with the lovastatin and AICAR combination provided greater protection of mitochondria/peroxisomes and myelin/axons, and greater improvement in clinical disease compared with individual drug treatments. In spinal cords of EAE mice, lovastatin-mediated inhibition of RhoA and AICAR-mediated activation of AMPK cooperatively enhanced the expression of the transcription factors and regulators (e.g. PPARα/ß, SIRT-1, NRF-1, and TFAM) required for biogenesis and the functions of mitochondria (e.g. OXPHOS, MnSOD) and peroxisomes (e.g. PMP70 and catalase). In summary, these studies document that oral medication with a combination of lovastatin and AICAR, which are individually known to have immunomodulatory effects, provides potent protection and repair of inflammation-induced loss and dysfunction of mitochondria and peroxisomes as well as myelin and axonal abnormalities in EAE. As statins are known to provide protection in progressive MS (Phase II study), these studies support that supplementation statin treatment with an AMPK activator may provide greater efficacy against MS.