Noncanonical Roles of Lipids in Different Cellular Fates.

Lipids are a diverse class of biomolecules. The biosynthesis and transport of these molecules are controlled by a considerable number of proteins, which facilitate spatiotemporal regulation of lipids during different fundamental cellular processes. Although lipids are traditionally considered as molecules for energy storage and as structural components of membranes, they are being increasingly recognized for their signaling roles. There is a growing appreciation of lipids' chemical diversity, which approaches that of proteins. In this Perspective, we discuss recent studies that suggest novel functions for distinct lipid species during different cellular processes. In particular, we discuss findings from our laboratory that illuminate the involvement of ceramides, polyunsaturated triacylglycerols, and very long chain fatty acids in different cellular fates. We also highlight recent innovative methods that have enabled the recognition of previously unknown lipid classes and/or roles of these molecules in different biological processes. We envision that advances in lipid identification, visualization, and perturbation will pave the way for broader investigations into this fascinating and influential class of biomolecules.

Regional but not global brain damage contributes to fatigue in multiple sclerosis.

PURPOSE: To use magnetic resonance (MR) imaging and advanced analysis to assess the role of lesions in normal-appearing white matter ( NAWM normal-appearing white matter ) and gray matter ( GM gray matter ) damage, global versus regional damage, and atrophy versus microstructural abnormalities in the pathogenesis of fatigue in multiple sclerosis ( MS multiple sclerosis ). MATERIALS AND METHODS: Local ethics committee approval and written informed consent were obtained. Dual-echo, double inversion-recovery, high-resolution T1-weighted and diffusion-tensor ( DT diffusion tensor ) MR was performed in 31 fatigued patients, 32 nonfatigued patients, and 35 control subjects. Global and regional atrophy and DT diffusion tensor MR measures of damage to lesions, NAWM normal-appearing white matter , and GM gray matter were compared (analysis of variance). RESULTS: Lesional, atrophy, and DT diffusion tensor MR measures of global damage to brain, white matter ( WM white matter ), and GM gray matter did not differ between fatigued and nonfatigued patients. Compared with nonfatigued patients and control subjects, fatigued patients experienced atrophy of the right side of the accumbens (mean volume ± standard deviation, 0.37 mL ± 0.09 in control subjects; 0.39 mL ± 0.1 in nonfatigued patients; and 0.33 mL ± 0.09 in fatigued patients), right inferior temporal gyrus ( ITG inferior temporal gyrus ) (Montreal Neurological Institute [ MNI Montreal Neurological Institute ] coordinates: 51, -51, -11; t value, 4.83), left superior frontal gyrus ( MNI Montreal Neurological Institute coordinates: -10, 49, 24; t value, 3.40), and forceps major ( MNI Montreal Neurological Institute coordinates: 11, -91, 18; t value, 3.37). They also had lower fractional anisotropy ( FA fractional anisotropy ) of forceps major ( MNI Montreal Neurological Institute coordinates: -17, -78, 6), left inferior fronto-occipital fasciculus ( MNI Montreal Neurological Institute coordinates: -25, 2, -11), and right anterior thalamic radiation ( ATR anterior thalamic radiation ) ( MNI Montreal Neurological Institute coordinates: 11, 2, -6) (P < .05, corrected). More lesions were found at T2-weighted imaging in fatigued patients. Multivariable model was used to identify right ITG inferior temporal gyrus atrophy (odds ratio, 0.83; 95% confidence interval [ CI confidence interval ]: 0.82, 0.97; P = .009) and right ATR anterior thalamic radiation FA fractional anisotropy (odds ratio, 0.74; 95% CI confidence interval : 0.61, 0.90; P = .003) as covariates independently associated with fatigue (C statistic, 0.85). CONCLUSION: Damage to strategic brain WM white matter and GM gray matter regions, in terms of microstructural abnormalities and atrophy, contributes to pathogenesis of fatigue in MS multiple sclerosis , whereas global lesional, WM white matter , and GM gray matter damage does not seem to have a role.

Changes of brain resting state functional connectivity predict the persistence of cognitive rehabilitation effects in patients with multiple sclerosis.

OBJECTIVE: We investigated whether the efficacy of 12-week cognitive rehabilitation in MS patients persists six months after treatment termination and, together with resting state (RS) functional connectivity (FC), changes on neuropsychological performance at follow-up. METHODS: Eighteen MS patients with cognitive deficits, assigned randomly either to undergo treatment (n=9) or not (n=9), underwent neuropsychological evaluation at baseline (t0), after 12 weeks of rehabilitation (t1) and at six-month follow-up (t2). RS fMRI was obtained at t0 and t1. Changes in neuropsychological performance and their correlations with RS FC modifications were assessed using longitudinal linear models. RESULTS: At t2 vs. t0, compared with the control group, treated group patients improved in tests of attention, executive function, depression and quality of life (QoL). Neuropsychological scores in these tests at t2 were significantly correlated with RS FC changes in cognitive-related networks and RS FC of the anterior cingulum. RS FC changes in the default mode network predicted cognitive performance and less severe depression, whereas RS FC changes of the executive network predicted better QoL. DISCUSSION: Changes in RS FC of cognitive-related networks helps to explain the persistence of the effects of cognitive rehabilitation after several months in relapsing-remitting multiple sclerosis patients and their improvement on depression and QoL scales.

Quantitative DigitoGraphy: a Comprehensive Real-Time Remote Monitoring System for Parkinson's Disease.

People with Parkinson's disease (PWP) face critical challenges, including lack of access to neurological care, inadequate measurement and communication of motor symptoms, and suboptimal medication management and compliance. We have developed QDG-Care: a comprehensive connected care platform for Parkinson's disease (PD) that delivers validated, quantitative metrics of all motor signs in PD in real time, monitors the effects of adjusting therapy and medication adherence and is accessible in the electronic health record. In this article, we describe the design and engineering of all components of QDG-Care, including the development and utility of the QDG Mobility and Tremor Severity Scores. We present the preliminary results and insights from the first at-home trial using QDG-Care. QDG technology has enormous potential to improve access to, equity of, and quality of care for PWP, and improve compliance with complex time-critical medication regimens. It will enable rapid "Go-NoGo" decisions for new therapeutics by providing high-resolution data that require fewer participants at lower cost and allow more diverse recruitment.

SREBP activation contributes to fatty acid accumulations in necroptosis.

Necroptosis is a type of programmed cell death. It is characterized by membrane permeabilization and is associated with the release of intracellular components due to compromised membrane integrity which induces a strong inflammatory response. We recently showed that the accumulation of very long chain fatty acids (VLCFAs) contributes to membrane permeabilization during necroptosis. However, the mechanisms that result in the accumulation of these cytotoxic lipids remain unknown. Using comparative transcriptomics and digital PCR validations, we found that several target genes of sterol regulatory element-binding proteins (SREBPs) were upregulated during necroptosis, suggesting that they might be responsible for the accumulation of VLCFA in this process. We demonstrated that activation of SREBPs during necroptosis exacerbates the permeability of the plasma membrane and cell death. Consistent with these observations, targeting sterol regulatory element-binding protein cleavage-activating protein (SCAP), a protein involved in SREBP activation, reversed the accumulation of VLCFAs, and restored cell death and membrane permeabilization during necroptosis. Collectively, our results highlight a role for SREBP in regulating lipid changes during necroptosis and suggest SREBP-mediated lipid remodeling as a potential target for therapeutics to reduce membrane permeabilization during necroptosis.

Prediction of subcutaneous drug absorption - Characterization of subcutaneous interstitial fluids as a basis for developing biorelevant in vitro models.

Unlike orally administered drugs, the absorption profile of subcutaneously injectable drugs in humans is difficult to predict from preclinical studies. Since the subcutaneous interstitial fluid (ISF) is the first fluid interacting with the administered formulation before the respective drug is absorbed, it could critically affect bioavailability. The aim of the present study was to gain a better understanding of the similarities and differences of ISF of different species. For this purpose, ISF was isolated from subcutaneous tissues of five preclinical animal species, i.e., mice, rats, minipig, landrace pig, non-human primates, and humans, using a centrifugation method, and characterized with respect to its major constituents and physicochemical properties. The results show trends between animal species, with ISF from non-human primates differing significantly from that of the other preclinical species for most parameters analyzed and showing similarities to ISF of human origin. Although from a statistical point of view it will be necessary to further increase the existing data sets, the presented data provide valuable information for the development of biorelevant in vitro models to predict the in vivo performance of subcutaneously administered formulations, as they provide fundamental information for the design of biorelevant ISF media for both preclinical species and humans.

Pet Presence Can Reduce Anxiety in the Elderly: The Italian Experience during COVID-19 Lockdown Assessed by an Electronic Survey.

The lockdown imposed in Italy due to the COVID-19 outbreak required restrictions that severely limited individual freedom to protect the population and reduce virus diffusion. This situation psychologically challenged the entire Italian population but mostly the elderly. The "Digital mental health approach" employs digital tools to evaluate and prevent increasing mental health problems. "Anonymous online electronic surveys" are digital tools that assess rates of mental health outcomes (using for example self-assessment/awareness tools). Immediately at the beginning of restrictions, we designed an electronic survey a) to remotely investigate the psychological impact of the lockdown and b) to compare the anxiety between pet owners and not-pet owners. A total of 3905 subjects filled out the survey; we focused our study on 781 (20%) elderly subjects. Dividing elderly patients between pet-owners (n = 405) and not-pet owners (n = 376), the pet owners showed a Zung scale score significantly lower in respect to the not-pet owners. We observed that, during the COVID-19 outbreak, the pet presence could have a positive effect on anxiety in the elderly subject. These results: (A) encourage the use of mobile technologies for the assessment of psychological disorders that can be promptly employed in emergencies such as the COVID-19 outbreak; (B) highlight the positive effect of pet interaction to mitigate the psychological distress in elderly people.

Protein acylation by saturated very long chain fatty acids and endocytosis are involved in necroptosis.

Necroptosis is a form of cell death characterized by receptor-interacting protein kinase activity and plasma membrane permeabilization via mixed-lineage kinase-like protein (MLKL). This permeabilization is responsible for the inflammatory properties of necroptosis. We previously showed that very long chain fatty acids (VLCFAs) are functionally involved in necroptosis, potentially through protein fatty acylation. Here, we define the scope of protein acylation by saturated VLCFAs during necroptosis. We show that MLKL and phosphoMLKL, key for membrane permeabilization, are exclusively acylated during necroptosis. Reducing the levels of VLCFAs decreases their membrane recruitment, suggesting that acylation by VLCFAs contributes to their membrane localization. Acylation of phosphoMLKL occurs downstream of phosphorylation and oligomerization and appears to be, in part, mediated by ZDHHC5 (a palmitoyl transferase). We also show that disruption of endosomal trafficking increases cell viability during necroptosis, possibly by preventing recruitment, or removal, of phosphoMLKL from the plasma membrane.

Membrane Disruption by Very Long Chain Fatty Acids during Necroptosis.

Necroptosis is a form of regulated cell death which results in loss of plasma membrane integrity, release of intracellular contents, and an associated inflammatory response. We previously found that saturated very long chain fatty acids (VLCFAs), which contain ≥20 carbons, accumulate during necroptosis. Here, we show that genetic knockdown of Fatty Acid (FA) Elongase 7 (ELOVL7) reduces accumulation of specific very long chain FAs during necroptosis, resulting in reduced necroptotic cell death and membrane permeabilization. Conversely, increasing the expression of ELOVL7 increases very long chain fatty acids and membrane permeabilization. In vitro, introduction of the VLCFA C24 FA disrupts bilayer integrity in liposomes to a greater extent than a conventional C16 FA. To investigate the microscopic origin of these observations, atomistic Molecular Dynamics (MD) simulations were performed. MD simulations suggest that fatty acids cause clear differences in bilayers based on length and that it is the interdigitation of C24 FA between the individual leaflets that results in disorder in the region and, consequently, membrane disruption. We synthesized clickable VLCFA analogs and observed that many proteins were acylated by VLCFAs during necroptosis. Taken together, these results confirm the active role of VLCFAs during necroptosis and point to multiple potential mechanisms of membrane disruption including direct permeabilization via bilayer disruption and permeabilization by targeting of proteins to cellular membranes by fatty acylation.

Apparent Splitting of S Waves Propagating Through an Isotropic Lowermost Mantle.

Observations of shear wave anisotropy are key for understanding the mineralogical structure and flow in the mantle. Several researchers have reported the presence of seismic anisotropy in the lowermost 150-250 km of the mantle (i.e., D '' layer), based on differences in the arrival times of vertically (SV) and horizontally (SH) polarized shear waves. By computing waveforms at a period > 6 s for a wide range of 1-D and 3-D Earth structures, we illustrate that a time shift (i.e., apparent splitting) between SV and SH may appear in purely isotropic simulations. This may be misinterpreted as shear wave anisotropy. For near-surface earthquakes, apparent shear wave splitting can result from the interference of S with the surface reflection sS. For deep earthquakes, apparent splitting can be due to the S wave triplication in D '' , reflections off discontinuities in the upper mantle, and 3-D heterogeneity. The wave effects due to anomalous isotropic structure may not be easily distinguished from purely anisotropic effects if the analysis does not involve full waveform simulations.

The role of pemetrexed in recurrent epithelial ovarian cancer: A scoping review.

Ovarian cancer is the leading cause of mortality among gynecologic malignancies, with most cases diagnosed at an advanced stage. Despite an initial response, most develop a recurrence and subsequent resistance to standard therapies. Pemetrexed (AlimtaTM) is a new generation multi-targeted antifolate initially approved for the treatment of malignant pleural mesothelioma. In recent years, it has shown promise in the treatment of recurrent epithelial ovarian cancer. In this review, we outline the current literature and discuss the future of pemetrexed in the setting of recurrent epithelial ovarian cancer.

Turning the Spotlight on Lipids in Non-Apoptotic Cell Death.

Although apoptosis has long dominated the spotlight, studies in the past two decades have expanded the repertoire of programmed cell death (PCD). Several forms of non-apoptotic regulated cell death have been identified, with important links to organismal homeostasis and different disease pathologies. Necroptosis, ferroptosis, pyroptosis, and NETosis are the major forms of PCD that have attracted attention. Clear biochemical distinctions differentiate these forms of non-apoptotic PCD at the protein and membrane levels. For instance, pore formation at the plasma membrane is a hallmark of necroptosis and pyroptosis; however, different proteins facilitate pore formation in these processes. Here, we will highlight the role of lipids in different forms of non-apoptotic PCD. In particular, we discuss how lipids can trigger or facilitate the membrane-related changes that result in cell death. We also highlight the use of small molecules in elucidating the mechanisms of non-apoptotic PCD and the potential of lipid biosynthetic pathways to perturb these processes for therapeutic applications as a future avenue of research.

Cholinesterase inhibition and alterations of hepatic metabolism by oral acute and repeated chlorpyrifos administration to mice.

Chlorpyrifos (CPF) is a broad spectrum organophosphorus insecticide bioactivated in vivo to chlorpyrifos-oxon (CPFO), a very potent anticholinesterase. A great majority of available animal studies on CPF and CPFO toxicity are performed in rats. The use of mice in developmental neurobehavioural studies and the availability of transgenic mice warrant a better characterization of CPF-induced toxicity in this species. CD1 mice were exposed to a broad range of acute (12.5-100.0mg/kg) and subacute (1.56-25mg/kg/day from 5 to 30 days) CPF oral doses. Functional and biochemical parameters such as brain and serum cholinesterase (ChE) and liver xenobiotic metabolizing system, including the biotransformation of CPF itself, have been studied and the no observed effect levels (NOELs) identified. Mice seem to be more susceptible than rats at least to acute CPF treatment (oral LD(50) 4.5-fold lower). The species-related differences were not so evident after repeated exposures. In mice a good correlation was observed between brain ChE inhibition and classical cholinergic signs of toxicity. After CPF-repeated treatment, mice seemed to develop some tolerance to CPF-induced effects, which could not be attributed to an alteration of P450-mediated CPF hepatic metabolism. CPF-induced effects on hepatic microsomal carboxylesterase (CE) activity and reduced glutathione (GSH) levels observed at an early stage of treatment and then recovered after 30 days, suggest that the detoxifying mechanisms are actively involved in the protection of CPF-induced effects and possibly in the induction of tolerance in long term exposure. The mouse could be considered a suitable experimental model for future studies on the toxic action of organophosphorus pesticides focused on mechanisms, long term and age-related effects.

Very Long Chain Fatty Acids Are Functionally Involved in Necroptosis.

Necroptosis is a form of regulated cell death that is linked to various human diseases. Distinct membrane-related, thus lipid-dependent, alterations take place during necroptosis. However, little is known about the roles of specific lipids in this process. We used an untargeted LC-MS-based approach to reveal that distinct lipid species are regulated at the molecular level during necroptosis. We found that ceramides and very long chain fatty acids accumulate during this process. Intrigued by the specificity of very long chain fatty acid accumulation, we focused on characterizing their involvement during necroptosis. Biochemical characterizations suggested that activated fatty acid biosynthesis and elongation could be responsible for these accumulations. We further showed that inhibition of fatty acid biosynthesis and depletion of very long chain fatty acids prevented loss of plasma membrane integrity and cell death, strongly suggesting that very long chain fatty acids are functionally involved in necroptosis.

Wet sol-gel derived silica for controlled release of proteins.

The potential of wet sol-gel derived silica gels as new matrices for the entrapment and sustained release of proteins was investigated. Model proteins, BSA, ribonuclease-A and avidin, with differing molecular weights and/or isoelectric points, were entrapped in two silica polymer formulations having different silica contents (4% and 12% wt/v). The conformational stability of the proteins after entrapment and their release after immersion into physiological conditions were measured. Circular dichroism analysis showed that protein conformation is maintained after entrapment and stability is enhanced. Protein-free formulations were injected intramuscularly into BALB/c mice to monitor the in vivo fate of the matrix, and the results showed that the gel is totally reabsorbed, without any apparent surrounding inflammation process. The time required for matrix bioerosion varied between one to three weeks, depending on its SiO(2) content. Erosion was also measured in vitro and the contribution of erosion and diffusion to the release of the embedded proteins was quantified. These data indicate that wet silica polymers obtained by the sol-gel route are promising matrices for the sustained release of protein drugs.

What providers want from the Primary Care Extension Service to facilitate practice transformation.

BACKGROUND AND OBJECTIVES: While several experts have shared their visions of the Primary Care Extension Service (PCES) as called for in the Affordable Care Act (ACA), little is known about providers' perspective. We aimed to identify the most and least desired resources that primary care providers want from the PCES. METHODS: A 70-question survey was administered to primary care providers (n=556) in Pennsylvania, one of four initial states chosen to develop the PCES infrastructure. Analysis focused on the highest and lowest ranked questions. RESULTS: The most desired PCES services include (1) identifying and coordinating mental health services, (2) improving office efficiency, (3) increasing overall revenues, and (4) strategies to help implement evidence-based clinical guidelines. The least desired PCES services include (1) implementing e-prescribing, (2) implementing an electronic medical record (EMR) system, (3) implementing group visits, (4) recruiting new patients, and (5) implementing open or advanced access scheduling. CONCLUSIONS: Despite expert models presented for the PCES, there is a critical need to ask primary care providers what they need from such a service. Our findings identified some divergences from key patient-centered medical home (PCMH) components, including the low ranking of services related to EMRs and increasing patient access. With interest growing in developing a PCES that would help spread innovation as outlined in the ACA, it's important to take a demand-side approach to the services providers most desire versus the more traditional supply-side approach that assumes the assistance providers need.

Functional correlates of preserved naming performance in amnestic Mild Cognitive Impairment.

Naming abilities are typically preserved in amnestic Mild Cognitive Impairment (aMCI), a condition associated with increased risk of progression to Alzheimer's disease (AD). We compared the functional correlates of covert picture naming and word reading between a group of aMCI subjects and matched controls. Unimpaired picture naming performance was associated with more extensive activations, in particular involving the parietal lobes, in the aMCI group. In addition, in the condition associated with higher processing demands (blocks of categorically homogeneous items, living items), increased activity was observed in the aMCI group, in particular in the left fusiform gyrus. Graph analysis provided further evidence of increased modularity and reduced integration for the homogenous sets in the aMCI group. The functional modifications associated with preserved performance may reflect, in the case of more demanding tasks, compensatory mechanisms for the subclinical involvement of semantic processing areas by AD pathology.

Virulence properties of type VII Streptococcus agalactiae (group B streptococci) and immunochemical analysis of capsular type polysaccharide.

Strains of a new polysaccharide type of group B streptococci (GBS), type VII, have been isolated from human carriers and invasive infections. Some of these strains bear the protein antigen c or R, as do other GBS serotypes. The capsular type polysaccharide is sialylated and this residue is involved in the immunodeterminant structure. All type VII strains examined were virulent in CD-1 mice; the LD50 after intraperitoneal (i.p.) challenge was 4.57 (SD 0.12) x10(7) cfu for the reference strain and 5.49 (SD 1.5) x10(7) cfu for clinical isolates. A particular feature of this serotype was the ability to induce septic arthritis not only when injected intravenously (i.v.), but also when injected i.p. Rabbit antiserum against the capsular type VII polysaccharide exhibited opsonic activity in a phagocytosis assay and protective activity against infection.

Simple and rapid technique for monitoring the quality of meningococcal polysaccharides by high performance size-exclusion chromatography.

The molecular size of meningococcal polysaccharides is an important physico-chemical parameter which correlates with immunogenicity. This paper describes the experimental conditions for high-performance size-exclusion chromatography on a PL Aquagel-OH 60 column to follow changes in the size distribution and therefore in the distribution coefficient (K(D)) of the meningococcal polysaccharides of groups A, C, Y and W-135 used to formulate anti-Neisseria meningitidis vaccines. The experimental conditions were also found to be suitable for a rapid monitoring of the quality (no group A polysaccharide depolymerization) of the tetravalent meningococcal polysaccharide vaccine.

Cognitive rehabilitation correlates with the functional connectivity of the anterior cingulate cortex in patients with multiple sclerosis.

We investigated how resting state (RS) functional connectivity (FC) of the anterior cingulate cortex (ACC) correlates with cognitive rehabilitation in relapsing remitting multiple sclerosis (RRMS) patients. A neuropsychological assessment and RS fMRI at baseline and after 12 weeks were obtained from 20 RRMS patients, who were assigned randomly to undergo treatment (n = 10) (treatment group-TG), which entailed computer-assisted cognitive rehabilitation of attention/information processing and executive functions for 3 days/week, or not to receive any cognitive rehabilitation (n = 10) (control group-CG). Voxel-wise changes of ACC RS FC were assessed using SPM8. In both groups, at the two study time points, ACC activity was correlated with the bilateral middle and inferior frontal gyrus, basal ganglia, posterior cingulate cortex, cerebellum, precuneus, middle temporal gyrus, and inferior parietal lobule (IPL). At follow up, compared to baseline, the TG showed an increased FC of the ACC with the right middle frontal gyrus (MFG) and right IPL, while the CG showed a decreased FC of the ACC with the right cerebellum and right inferior temporal gyrus (ITG). A significant "treatment × time" interaction was found for the increased FC of the right IPL and for the decreased FC of the right ITG. In the TG only, significant correlations (p < 0.001) were found between improvement of PASAT performance and RS FC of the ACC with the right MFG (r = 0.88) and right IPL (r = 0.76). In MS, cognitive rehabilitation correlates with changes in RS FC of brain regions subserving the trained functions. fMRI might be useful to monitor rehabilitative strategies in MS.