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Objectives: In skeletal muscle, the capillaries have tight junctions (TJs) that are structurally similar to those in the blood‐brain barrier (BBB) and blood‐nerve barrier (BNB). Although the endomysial capillaries include endothelial cells and pericytes, the properties of the endomysial pericytes have not been well elucidated. The purpose of this study was to establish a human skeletal muscle pericyte cell (HSMPCT) line, and examine whether the established cell line has similar properties to pericytes from the BBB and BNB. Method: HSMPCTs were isolated and introduced with retroviruses harboring temperature‐sensitive SV40 T antigen and telomerase genes. Then, we examined whether this cell line modifies the barrier function of a human skeletal muscle microvascular endothelial cell line, TSM15. Results: A new HSMPCT cell line was successfully established. The conditioned media (CM) from HSMPCTs increased the transendothelial electrical resistance of the TSM15 cells and decreased permeability to 10kD‐dextran. Glial cell line‐derived neurotrophic factor (GDNF) and basic fibroblast growth factor (bFGF) increased the barrier function of TSM15 cells. The CM of HSMPCTs with anti‐GDNF or anti‐bFGF antibodies attenuated the effect of upregulating the barrier function of TSM15 cells. These results indicate that soluble factors, such as GDNF and bFGF, released from HSMPCTs strengthen the barrier function of TSM15 cells. Conclusion: GDNF and bFGF released from HSMPCTs increase the barrier function of endomysial endothelium. These HSMPCTs and TSM15 cells, which are recognized as an in vitro model of the endomysial capillary, might facilitate the analysis of the pathophysiology of the microvasculature in human skeletal muscle. [ABSTRACT FROM AUTHOR]

Delirium is a syndrome that can arise from many causes or underlying conditions, and though it has been reported in younger patients, it is more prevalent in older people, though it can occur in other age groups as well. Identifying delirium is challenging in older people because of the coexistence of underlying dementia or depression, which may further complicate the presentation. Drug-induced delirium is one of the major causes of delirium, and evaluation of this potential cause or contribution is an important component of the evaluation process, since it can lead to poor patient outcomes. Part one of this three part series reviewed the epidemiology, pathophysiology, evaluation, diagnostic process, and causes of delirium in older people, with a focus on the pharmacological causes. Part two of this series continues to review drugs and drug classes that can cause or contribute to delirium in older people. [ABSTRACT FROM AUTHOR]

Objective: To assess the utility of the preoperative Sarcopenia index (SI) as a predictive marker of the risk of postoperative complications following hip fracture surgery in older adults.Study Design: This observational study enrolled older adults with hip fracture who were hospitalized in the Department of Orthopedics of West China Hospital, Sichuan University, from December 7, 2010 - June 14, 2017, and who underwent hip fracture surgery.Primary Outcome and Measures: Clinical data were collected from medical records and serum creatinine and cystatin C were measured before surgery. Outcomes included postoperative complications such as pneumonia, urinary tract infection, respiratory failure, heart failure, and non-grade A healing. Binary logistic regression analyses were used to analyze association between SI and postoperative complications.Results: A total of 897 patients aged 60 years and over were enrolled in this study (age range: 60 - 100 years), of whom 306(34.1%)were male, and 591(65.9%)were female. Postoperative complications included pneumonia (12%), urinary tract infections (1.8%), respiratory failure (1.5%), heart failure (1.6%), and non-A- grade healing (3.6%). In the patient group that received joint replacements, the incidence of pneumonia was negatively associated with SI values. After adjusting for potential confounding factors, binary logistic regression analyses showed that a higher SI was independently associated with a lower risk of pneumonia after joint replacement surgery (OR:0.39, 95% CI:0.18-0.89, P<0.05). However, we did not find statistically significant association between SI and the risk of postoperative complications other than pneumonia among patients with two types of hip fracture surgery.Conclusion: The SI based on serum creatinine and cystatin C can predict pneumonia rather than other postoperative complications among older patients with hip fracture after joint replacement surgery. [ABSTRACT FROM AUTHOR]

Several neurological and psychiatric disorders have been associated with impairments in GABAergic inhibitory neurons in the brain. Thus, in the current era of accelerated development of molecular medicine and biologically-based drugs, there is a need to identify gene regulatory sequences that can be utilized for selectively manipulating the expression of nucleic acids and proteins in GABAergic neurons. This is particularly important for the use of viral vectors in gene therapy. In this Mini Review, we discuss the use of various gene regulatory elements for targeting GABAergic neurons, with an emphasis on adeno-associated viral vectors, the most widely used class of viral vectors for treating brain diseases. [ABSTRACT FROM AUTHOR]

This commentary provides an overview of the putamen as an established target site for gene therapy in treating aromatic l‐amino acid decarboxylase (AADC) deficiency and Parkinson's disease, two debilitating neurological disorders that involve motor dysfunction caused by dopamine deficiencies. The neuroanatomy and the function of the putamen in motor control provide good rationales for targeting this brain structure. Additionally, the efficacy and safety of intraputaminal gene therapy demonstrate that restoration of dopamine synthesis in the putamen by using low doses of adeno‐associated viral vector serotype 2 to deliver the hAADC gene is well tolerated. This restoration leads to sustained improvements in motor and nonmotor symptoms of AADC deficiency and improved uptake and conversion of exogenous l‐DOPA into dopamine in Parkinson's patients. [ABSTRACT FROM AUTHOR]

Dynamic molecular processes, such as short- or long-range proton (H+) and ion (M+) motions, and molecular rotations in electrical conducting and magnetic molecular assemblies enable the fabrication of electron--H+ (or M+) coupling systems, while crystal lattice dynamics and molecular conformation changes in hydrogen-bonded molecular crystals have been utilised in external stimuli responsive reversible gas-induced gate opening and molecular adsorption/desorption behavior. These dynamics of the polar structural units are responsible for the dielectric measurements. The H+ dynamics are formed from ferroelectrics and H+ conductors, while the dynamic M+ motions of Li+ and Na+ involve ionic conductors and coupling to the conduction electrons. In n-type organic semiconductors, the crystal lattices are modulated by replacing M+ cations, with cations such as Li+, Na+, K+, Rb+, and Cs+. The use of polar rotator or inversion structures such as alkyl amides, m-fluoroanilinium cations, and bowl-shaped trithiasumanene p-cores enables the formation of ferroelectric molecular assemblies. The host--guest molecular systems of ESIPT fluorescent chromic molecules showed interesting molecular sensing properties using various bases, where the dynamic transformation of the crystal lattice and the molecular conformational change were coupled to each other. [ABSTRACT FROM AUTHOR]

Knowledge regarding the relationship between muscles and the corresponding motor neurons would allow therapeutic genes to transport into specific spinal cord segments. Retrograde tracing technique by targeting the motor endplate (MEP), a highly specialized structure that offers direct access to the spinal motor neurons, has been used to elucidate the connectivity between skeletal muscles and the innervating motor neuron pools. However, current injection strategies mainly based on blind injection or the local MEP region might lead to an underestimation of the motor neuron number due to the uneven distribution of MEP in skeletal muscles. In this work, we proposed a novel intramuscular injection strategy based on the 3D distribution of the MEPs in skeletal muscles, applied the 3D intramuscular injection to the gastrocnemius and tibialis anterior for retrograde tracing of the corresponding motor neurons, and compared this with the existing injection strategy. The intramuscular diffusion of the tracer demonstrated that 3D injection could maximize the retrograde transport by ensuring a greater uptake of the tracer by the MEP region. In combination with optical clearing and imaging, we performed 3D mapping and quantification of the labeled motor neurons and confirmed that 3D injection could label more motor neurons than the current injection method. It is expected that 3D intramuscular injection strategy will help elucidate the connective relationship between muscles and motor neurons faithfully and becomes a promising tool in the development of gene therapy strategies for motor neuron diseases. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is one of the most common neurodegenerative diseases, defined as motor and non-motor symptoms associated with the loss of dopaminergic neurons and a decreased release of dopamine (DA). Currently, PD patients are believed to have a neuropathological basis denoted by the presence of Lewy bodies (LBs) or Lewy neurites (LNs), which mostly comprise α-synuclein (α-syn) inclusions. Remarkably, there is a growing body of evidence indicating that the inclusions undergo template-directed aggregation and propagation via template-directed among the brain and peripheral organs, mainly in a prion-like manner. Interestingly, some studies reported that an integral loop was reminiscent of the mechanism of Parkinson's disease, denoting that α-syn as prionoid was transmitted from the periphery to the brain via specific pathways. Also the systematic life cycle of α-syn in the cellular level is illustrated. In this review, we critically assess landmark evidence in the field of Parkinson's disease with a focus on the genesis and prion-like propagation of the α-syn pathology. The anatomical and cell-to-cell evidences are discussed to depict the theory behind the propagation and transferred pathways. Furthermore, we highlight effective therapeutic perspectives and clinical trials targeting prion-like mechanisms. Major controversies surrounding this topic are also discussed. [ABSTRACT FROM AUTHOR]

Single molecules that co-transport cations as well as anions across lipid membranes are few despite their high biological utility. The elegant yet simple lipidomimmetic peptide design herein enables efficient HCl transport without the use of any external additives for proton transport. The carboxylic acids in the dipeptide scaffold provide a handle to append two long hydrophobic tails and also provide a polar hydrophilic carboxylate group. The peptide central unit also provides NH sites for anion binding. Protonation of the carboxylate group coupled with the weak halide binding of the terminal NH group results in HCl transport with transport rates of H + >Cl - . The lipid-like structure also facilitates seamless membrane integration and flipping of the molecule. The biocompatibility, design simplicity, and potential pH regulation of these molecules open up several avenues for their therapeutic use., (© 2023 Wiley-VCH GmbH.)

In this work, it was first found that tectoridin can bind to the fat mass and obesity-associated protein (FTO) by isothermal titration calorimetry (ITC). Spontaneous exothermic reactions occurred between tectoridin and FTO driven by entropy. A static quenching mechanism was developed for this interaction by using the spectroscopic technique. Results of both ITC and fluorescence spectroscopy indicated that hydrophobic interactions are predominant in the formation of the complex. The binding parameters obtained from ITC are comparable to those obtained from fluorescence spectroscopy. The inhibition of tectoridin on the demethylase activity of FTO was confirmed by enzymatic activity assays. This study will provide the basis for the development of inhibitors for FTO demethylase. [ABSTRACT FROM AUTHOR]

We show that multipodal polycationic receptors function as anion‐responsive light‐emitters in water. Prevailing paradigms utilize rigid holes and cavities for ion recognition. We instead built open amphiphilic scaffolds that trigger polar‐to‐nonpolar environment transitions around cationic fluorophores upon anion complexation. This ion‐pairing and aggregation event produces a dramatic enhancement in the emission intensity, as demonstrated by perchlorate as a non‐spherical hydrophobic anion model. A synergetic interplay of C−H⋅⋅⋅anion hydrogen bonding and tight anion–π+ contacts underpins this supramolecular phenomenon. By changing the aliphatic chain length, we demonstrate that the response profile and threshold of this signaling event can be controlled at the molecular level. With appropriate molecular design, inherently weak, ill‐defined, and non‐directional van der Waals interaction enables selective, sensitive, and tunable recognition in water. [ABSTRACT FROM AUTHOR]

Inspired by calcium-induced reversible assembly and disassembly of membrane proteins found in nature, here we developed a phosphorylated amphiphile (PA) that contains an oligo(phenylene–ethynylene) unit as a hydrophobic unit and a phosphate ester group as a hydrophilic calcium-binding unit. We demonstrated that PA can assemble and disassemble in a reversible manner in response to the sequential addition of calcium chloride and ethylene-diaminetetraacetic acid within the lipid bilayer membranes for the first time as a synthetic molecule. [ABSTRACT FROM AUTHOR]

Background: The simultaneous assessment of visceral adiposity and muscle mass might be useful to monitor the risk of non-alcoholic fatty liver disease (NAFLD) progression in large population. We aimed to investigate the value of serum creatinine-to-cystatin C ratio (CCR) in evaluating these two parameters and predicting liver steatosis and fibrosis. Methods: 154 overweight/obese inpatients (49 males, 105 females) scheduled for bariatric surgery and 49 non-overweight/obese volunteers (18 males, 31 females) responded to the hospital advertisement were involved in the cross-sectional study. Liver steatosis and fibrosis were diagnosed with transient elastography (TE). The psoas muscle area (PMA) and visceral fat area (VFA) were measured using magnetic resonance imaging. Results: The body mass index, insulin resistance, and lipid profiles showed significant differences between the CCR tertiles. Multiple regression analyses revealed that the CCR was significantly associated with the controlled attenuation parameter (β = −0.30, P = 0.006 in males; β = −0.19, P = 0.017 in females) and liver stiffness measurements in males (β = −0.246, P = 0.044). A low CCR was associated with moderate-to-severe steatosis (P < 0.001), significant liver fibrosis (P < 0.01), and excellent predictive power for these two conditions (P < 0.01). The CCR had a negative correlation with the VFA/PMA ratio (r = −0.584, P < 0.001 in males; r = −0.569, P < 0.001 in females). Conclusions: The CCR is a serum marker for muscle-adjusted visceral fat mass, and a low CCR is associated with an increased risk of progressive NAFLD. [ABSTRACT FROM AUTHOR]

Introduction: We sought to provide an overview of the published and currently ongoing movement disorders clinical trials employing gene therapy, defined as a technology aiming to modulate the expression of one or more genes to achieve a therapeutic benefit. Methods: We systematically reviewed movement disorders gene therapy clinical trials from PubMed and ClinicalTrials.gov using a searching strategy that included Parkinson disease (PD), Huntington disease (HD), amino acid decarboxylase (AADC) deficiency, multiple system atrophy (MSA), progressive supranuclear palsy (PSP), dystonia, tremor, ataxia, and other movement disorders. Data extracted included study characteristics, investigational product, route of administration, safety/tolerability, motor endpoints, and secondary outcomes (i.e., neuroimaging, biomarkers). Results: We identified a total of 46 studies focusing on PD (21 published and nine ongoing), HD (2 published and 5 ongoing), AADC deficiency (4 published and 2 ongoing), MSA (2 ongoing), and PSP (1 ongoing). In PD, intraparenchymal infusion of viral vector-mediated gene therapies demonstrated to be safe and showed promising preliminary data in trials aiming at restoring the synthesis of dopamine, enhancing the production of neurotrophic factors, or modifying the functional interaction between different nodes of the basal ganglia. In HD, monthly intrathecal delivery of an antisense oligonucleotide (ASO) targeting the huntingtin protein (HTT) mRNA proved to be safe and tolerable, and demonstrated a dose-dependent reduction of the cerebrospinal fluid levels of mutated HTT, while a small phase-I study testing implantable capsules of cells engineered to synthesize ciliary neurotrophic factor failed to show consistent drug delivery. In AADC deficiency, gene replacement studies demonstrated to be relatively safe in restoring catecholamine and serotonin synthesis, with promising outcomes. Ongoing movement disorders clinical trials are focusing on a variety of gene therapy approaches including alternative viral vector serotypes, novel recombinant genes, novel delivery techniques, and ASOs for the treatment of HD, MSA, and distinct subtypes of PD (LRRK2 mutation or GBA1 mutation carriers). Conclusion: Initial phase-I and -II studies tested the safety and feasibility of gene therapy in PD, HD, and AADC deficiency. The ongoing generation of clinical trials aims to test the efficacy of these approaches and explore additional applications for gene therapy in movement disorders. [ABSTRACT FROM AUTHOR]

At present there is a significant unmet need for clinically available treatments for Parkinson's disease (PD) patients to stably restore balance to dopamine network function, leaving patients with inadequate management of symptoms as the disease progresses. Gene therapy is an attractive approach to impart a durable effect on neuronal function through introduction of genetic material to reestablish dopamine levels and/or functionally recover dopaminergic signaling by improving neuronal health. Ongoing clinical gene therapy trials in PD are focused on enzymatic enhancement of dopamine production and/or the restoration of the nigrostriatal pathway to improve dopaminergic network function. In this review, we discuss data from current gene therapy trials for PD and recent advances in study design and surgical approaches. [ABSTRACT FROM AUTHOR]

ABCB1, also called MDR1 or P‐glycoprotein, exports various hydrophobic compounds and plays an essential role as a protective physiological barrier in several organs, including the brain, testis, and placenta. However, little is known about the structural mechanisms that allow ABCB1 to recognize hydrophobic compounds of diverse structures or the coupling of ATP hydrolysis to uphill substrate export. High‐resolution X‐ray crystal structures of the pre‐ and post‐transport states and FRET analyses in living cells have revealed that an aromatic hydrophobic network at the top of the inner cavity is key for the conformational change in ABCB1 that is triggered by a hydrophobic substrate. ATP binding, but not hydrolysis, induces a progressive network that results in a twisting motion of the whole protein, squeezing out the substrate directly to the extracellular space. This twist‐and‐squeeze mechanism by which ABCB1 exports hydrophobic substrates is distinct from those of other transporters. [ABSTRACT FROM AUTHOR]

Background: Response to thymectomy in myasthenia gravis (MG) is influenced by various patient-, disease-, and therapy-related factors.Methods: Retrospective analysis of 128 patients with MG who underwent maximal thymectomy over 15 years was done to identify the determinants of suboptimal clinical outcome.Results: Among the 128 patients, 62 (48.4%) were females with a mean age of 38.97 (12.29) years. Thymomatous MG occurred in 66 (51.6%). Overall improvement from preoperative status was noted in 88 (68.8%) patients after mean follow-up of 51.68 (33.21) months. The presence of thymoma was the major predictor of suboptimal clinical outcome (P = 0.001), whereas age, gender, preoperative disease severity, and seropositive status did not attain significance. Patients with better outcome had received higher steroid dose preoperatively (P = 0.035).Conclusions: Suboptimal response after thymectomy occurred in one-third of MG patients, more commonly with thymomatous MG. Relationship of preoperative steroid therapy to remission merits evaluation. [ABSTRACT FROM AUTHOR]

The study aimed to investigate the influence of shrunken pore syndrome (SPS), defined as a cystatin C (CysC)-based estimated glomerular filtration rate (eGFRCysC) <60% of the creatinine (Cr)-based eGFR (eGFRCr), on bone mineral density (BMD) in patients with rheumatic diseases. A total of 831 patients with rheumatic diseases were enrolled in the study. Patients were classified into the SPS group (G-SPS) and non-SPS group (G-nSPS). The correlation between the presence of SPS and BMD of the lumbar spine (BMD_LS), BMD of the femoral neck (BMD_FN), serum parathyroid hormone (PTH) level, chronic kidney dysfunction (CKD), and parameters were evaluated statistically. The prevalence of SPS was 4.0%. Serum PTH level, tartrate-resistant acid phosphatase-5b (TRACP-5b), and eGFRCr in the G-SPS were significantly higher than in the G-nSPS, whereas BMD_LS and BMD_FN in the G-SPS were significantly lower than in the G-nSPS. Serum PTH level was significantly correlated with CysC. BMD_LS had no significant correlation with BMD_FN. The presence of SPS was the only factor that demonstrated significant negative correlation with both BMD_LS and BMD_FN. Relationship between BMD_LS and the presence of SPS was present regardless of CKD stage; however, the negative relationship between BMD_LS and serum PTH was observed only in CKD stage 1 and 2 patients. BMD_FN demonstrated significant negative correlation with serum PTH in the group with progression of CKD. These results suggest that there is a serious potential risk of osteoporosis in patients with SPS and increased PTH, and BMD_LS poses a higher risk in CKD stage 1 and 2. [ABSTRACT FROM AUTHOR]

Decades of preclinical and clinical studies developing gene therapy for hemophilia are poised to bear fruit with current promising pivotal studies likely to lead to regulatory approval. However, this recent success should not obscure the multiple challenges that were overcome to reach this destination. Gene therapy for hemophilia A and B benefited from advancements in the general gene therapy field, such as the development of adeno-associated viral vectors, as well as disease-specific breakthroughs, like the identification of B-domain deleted factor VIII and hyperactive factor IX Padua. The gene therapy field has also benefited from hemophilia B clinical studies, which revealed for the first time critical safety concerns related to immune responses to the vector capsid not anticipated in preclinical models. Preclinical studies have also investigated gene transfer approaches for other rare inherited bleeding disorders, including factor VII deficiency, von Willebrand disease, and Glanzmann thrombasthenia. Here we review the successful gene therapy journey for hemophilia and pose some unanswered questions. We then discuss the current state of gene therapy for these other rare inherited bleeding disorders and how the lessons of hemophilia gene therapy may guide clinical development. [ABSTRACT FROM AUTHOR]

A series of peptide-appended bisresorcinarenes were synthesized, which adopted tubular conformation induced by intramolecular hydrogen bonds. The derivatives formed unimolecular artificial transmembrane channels in lipid bilayers to enable selective transport of monovalent cations. Importantly, the channels exhibited reversible ligand-gating behavior in response to alkyl amine and Cu2+. [ABSTRACT FROM AUTHOR]

Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare genetic disorder belonging to the group of vacuolating leukodystrophies. It is characterized by megalencephaly, loss of motor functions, epilepsy, and mild mental decline. In brain biopsies of MLC patients, vacuoles were observed in myelin and in astrocytes surrounding blood vessels. It is mainly caused by recessive mutations in MLC1 and HEPACAM (also called GLIALCAM) genes. These disease variants are called MLC1 and MLC2A with both types of patients sharing the same clinical phenotype. Besides, dominant mutations in HEPACAM were also identified in a subtype of MLC patients (MLC2B) with a remitting phenotype. MLC1 and GlialCAM proteins form a complex mainly expressed in brain astrocytes at the gliovascular interface and in Bergmann glia at the cerebellum. Both proteins regulate several ion channels and transporters involved in the control of ion and water fluxes in glial cells, either directly influencing their location and function, or indirectly regulating associated signal transduction pathways. However, the MLC1/GLIALCAM complex function and the related pathological mechanisms leading to MLC are still unknown. It has been hypothesized that, in MLC, the role of glial cells in brain ion homeostasis is altered in both physiological and inflammatory conditions. There is no therapy for MLC patients, only supportive treatment. As MLC2B patients show an MLC reversible phenotype, we speculated that the phenotype of MLC1 and MLC2A patients could also be mitigated by the re-introduction of the correct gene even at later stages. To prove this hypothesis, we injected in the cerebellar subarachnoid space of Mlc1 knockout mice an adeno-associated virus (AAV) coding for human MLC1 under the control of the glial-fibrillary acidic protein promoter. MLC1 expression in the cerebellum extremely reduced myelin vacuolation at all ages in a dose-dependent manner. This study could be considered as the first preclinical approach for MLC. We also suggest other potential therapeutic strategies in this review. [ABSTRACT FROM AUTHOR]

Transmembrane anion transport is an important biological process in maintaining cellular functions. Thus, synthetic anion transporters are widely developed for their biological applications. Imidazolinium was introduced as anion recognition site to a multiblock amphiphilic structure that consists of octa(ethylene glycol) and aromatic units. Ion transport assay using halide‐sensitive lucigenin and pH‐sensitive 8‐hydroxypyrene‐1,3,6‐trisulfonate (HPTS) revealed that imidazolinium‐based multiblock amphiphile (IMA) transports anions and showed high selectivity for nitrate, which plays crucial roles in many biological events. Temperature‐dependent ion transport assay using 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine (DPPC) indicated that IMA works as a mobile carrier. 1H NMR titration experiments indicated that the C2 proton of the imidazolinium ring recognizes anions via a (C−H)+⋅⋅⋅X− hydrogen bond. Furthermore, all‐atom molecular dynamics simulations revealed a dynamic feature of IMA within the membranes during ion transportation. [ABSTRACT FROM AUTHOR]

Gene therapy to treat pharmacoresistant temporal lobe epilepsy in humans is now being developed using an AAV vector (CG01) that encodes the combination of neuropeptide Y and its antiepileptic receptor Y2. With this in mind, the present study aimed to provide important preclinical data on the effects of CG01 on the duration of transgene expression, cellular tropism, and potential side effects on body weight and cognitive function. The CG01 vector was administered unilaterally into the dorsal and ventral hippocampus of adult male rats and expression of both transgenes was found to remain elevated without a sign of decline at 6 months post-injection. CG01 appeared to mediate expression selectively in hippocampal neurons, without expression in astrocytes or oligodendrocytes. No effects were seen on body weight as well as on short- or long-term memory as revealed by testing in the Y-maze or Morris water maze tests. Thus these data show that unilateral CG01 vector treatment as future gene therapy in pharmacoresistant temporal lobe epilepsy patients should result in stable and long-term expression predominantly in neurons and be well tolerated without side effects on body weight and cognitive function. [ABSTRACT FROM AUTHOR]

ATP‐binding cassette (ABC) transporters are fascinating molecular machines that are capable of transporting a large variety of chemically diverse compounds. The energy required for translocation is derived from binding and hydrolysis of ATP. All ABC transporters share a basic architecture and are composed of two transmembrane domains and two nucleotide binding domains (NBDs). The latter harbor all conserved sequence motifs that hallmark the ABC transporter superfamily. The NBDs form the nucleotide binding sites (NBSs) in their interface. Transporters with two active NBSs are called canonical transporters, while ABC exporters from eukaryotic organisms, including humans, frequently have a degenerate NBS1 containing noncanonical residues that strongly impair ATP hydrolysis. Here, we summarize current knowledge on degenerate ABC transporters. By integrating structural information with biophysical and biochemical evidence of asymmetric function, we develop a model for the transport cycle of degenerate ABC transporters. We will elaborate on the unclear functional advantages of a degenerate NBS. [ABSTRACT FROM AUTHOR]

The human multidrug transporter ABCG2 is required for physiological detoxification and mediates anticancer drug resistance. Here, we identify pivotal residues in the first intracellular loop (ICL1), constituting an intrinsic part of the transmission interface. The architecture includes a triple helical bundle formed by the hot spot helix of the nucleotide‐binding domain, the elbow helix, and ICL1. We show here that the highly conserved ICL1 residues G462, Y463, and Y464 are essential for the proper cross talk of the closed nucleotide‐binding domain dimer with the transmembrane domains. Hence, ICL1 acts as a molecular spring, triggering the conformational switch of ABCG2 before substrate extrusion. These data suggest that the ABCG2 transmission interface may offer therapeutic options for the treatment of drug‐resistant malignancies. [ABSTRACT FROM AUTHOR]

ATP‐binding cassette (ABC) proteins play diverse roles in all living organisms, making them an attractive model for evolution. Early evolution of ancestral unicellular organisms entailed the acquisition of at least three types of ABC proteins: type 1 ABC proteins to import nutrients, and type 2 and 3 ABC proteins to generate the outer cell membrane by flopping and loading lipids onto acceptors, respectively. To export various toxic lipophilic compounds, cells evolutionarily acquired a fourth type of ABC protein. This suggests that ABC proteins may have played an important role in evolution, especially when life became terrestrial, protecting plants and animals from water loss and pathogen infection. ABC proteins are also assumed to have accelerated the evolution of vertebrates by allowing cholesterol to function for intramembrane signaling. In this review, we discuss the roles of ABC proteins in the evolution of bacteria, plants, and animals. [ABSTRACT FROM AUTHOR]

Structural data on ABCG5/G8 and ABCG2 reveal a unique molecular architecture for subfamily G ATP‐binding cassette (ABCG) transporters and disclose putative substrate‐binding sites. ABCG5/G8 and ABCG2 appear to use several unique structural motifs to execute transport, including the triple helical bundles, the membrane‐embedded polar relay, the re‐entry helices, and a hydrophobic valve. Interestingly, ABCG2 shows extreme substrate promiscuity, whereas ABCG5/G8 transports only sterol molecules. ABCG2 structures suggest a large internal cavity, serving as a binding region for substrates and inhibitors, while mutational and pharmacological analyses support the notion of multiple binding sites. By contrast, ABCG5/G8 shows a collapsed cavity of insufficient size to hold substrates. Indeed, mutational analyses indicate a sterol‐binding site at the hydrophobic interface between the transporter and the lipid bilayer. In this review, we highlight key differences and similarities between ABCG2 and ABCG5/G8 structures. We further discuss the relevance of distinct and shared structural features in the context of their physiological functions. Finally, we elaborate on how ABCG2 and ABCG5/G8 could pave the way for studies on other ABCG transporters. [ABSTRACT FROM AUTHOR]

Serum creatinine (Cr) is a biosynthetic product of creatine phosphate metabolism in muscles and is closely related to total muscle mass, but it is not easily affected by diet. Several studies have tried to explore the role of serum Cr levels in amyotrophic lateral sclerosis (ALS), but the results were inconsistent. Therefore, our study aims to explore the differences of serum Cr levels between ALS patients and controls and whether serum Cr at baseline is an independent predictor of survival. Methods: We searched all the related studies that probed into the association between Serum Cr levels and ALS based on PubMed, EMBASE and Cochrane library from October 1952 to February 2019. The quality of the included studies was evaluated by using Newcastle-Ottawa Scale (NOS), and all the statistical analysis of this meta-analysis was performed by Stata version 12.0. Results: Eight studies with a total of 11377 ALS patients and 937 controls were included. Among them, five studies indicated that ALS patients had lower serum Cr levels (SMD = −0.78, 95%CI [−0.97, −0.60]) compared to controls, and three studies showed that higher serum Cr levels in ALS patients were related to lower overall mortality (HR 0.89, 95%CI [0.80, 0.99]). Conclusion: The levels of serum Cr in ALS patients are significantly lower than those in controls, and they are inversely related to overall mortality in ALS patients. Therefore, the serum Cr, an easily accessible serological factor, may serve as a prognostic biomarker. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a complex, multisystem, progressive, degenerative disorder characterized by severe, debilitating motor dysfunction, cognitive impairments, and mood disorders. Although preclinical research has traditionally focused on the motor deficits resulting from the loss of nigrostriatal dopaminergic neurons, up to two thirds of PD patients present separate and distinct behavioral changes. Loss of basal forebrain cholinergic neurons occurs as early as the loss of dopaminergic cells and contributes to the cognitive decline in PD. In addition, attentional deficits can limit posture control and movement efficacy caused by dopaminergic cell loss. Complicating the picture further is intracellular α-synuclein accumulation beginning in the enteric nervous system and diffusing to the substantia nigra through the dorsal motor neurons of the vagus nerve. It seems that α-synuclein's role is that of mediating dopamine synthesis, storage, and release, and its function has not been completely understood. Treating a complex, multistage network disorder, such as PD, likely requires a multipronged approach. Here, we describe a few approaches that could be used alone or perhaps in combination to achieve a greater mosaic of behavioral benefit. These include (1) using encapsulated, genetically modified cells as delivery vehicles for administering neuroprotective trophic factors, such as GDNF, in a direct and sustained means to the brain; (2) immunotherapeutic interventions, such as vaccination or the use of monoclonal antibodies against aggregated, pathological α-synuclein; (3) the continuous infusion of levodopa-carbidopa through an intestinal gel pad to attenuate the loss of dopaminergic function and manage the motor and non-motor complications in PD patients; and (4) specific rehabilitation treatment programs for drug-refractory motor complications. [ABSTRACT FROM AUTHOR]

The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned common marmoset has been used extensively to model Parkinson's disease, l-3,4-dihydroxyphenylalanine (l-DOPA)-induced dyskinesia and, more recently, dopaminergic psychosis. Whereas several experimental drugs have been tested in this primate, many of which subsequently underwent clinical trials, efficacy thresholds in the marmoset that would predict efficacy in the clinic are lacking. Here, we aimed to determine such efficacy end points that would be indicative of likely efficacy in clinical studies. To do so, we used the evidence-based medicine reviews published by the International Parkinson and Movement Disorder Society (IPMDS) to select drugs that were rated as clinically efficacious, likely efficacious or not efficacious for the treatment of parkinsonism, dyskinesia and psychosis. We then reviewed the literature in the MPTP-lesioned marmoset and identified articles reporting the effects of drugs that were included in the IPMDS recommendations, following which we estimated efficacy thresholds in the marmoset that would predict efficacy at the clinical level. We propose that, when drugs are administered as monotherapy, ≥ 50% reduction of global parkinsonism may be necessary to predict the possibility of clinical efficacy. As adjunct to a low dose of l-DOPA, we propose that an additional reduction of global parkinsonism ≥ 25% might predict clinical efficacy. As adjunct to an optimal dose of l-DOPA, we propose that additional anti-parkinsonian benefit ≥ 20%, with global parkinsonism as the end point, might predict clinical efficacy. For the treatment of dyskinesia, we suggest that the predictability threshold be set at ≥ 25% reduction of peak dose dyskinesia, while we believe that this threshold should be > 50% reduction of peak dose psychosis-like behaviours for psychosis-related end points. This article represents the first step in determining what efficacy might be necessary to achieve in pre-clinical studies in the MPTP-lesioned marmoset prior to confidently advancing drugs to clinical trials. We hope that it will help in the drug discovery and development process, notably by avoiding exposing patients to drugs that have little probability of clinical efficacy based upon pre-clinical experiments. [ABSTRACT FROM AUTHOR]

Optogenetics is an invasive neuromodulation technology involving the use of light to control the activity of individual neurons. Even though optogenetics is a relatively new neuromodulation tool whose various implications have not yet been scrutinized, it has already been approved for its first clinical trials in humans. As optogenetics is being intensively investigated in animal models with the aim of developing novel brain stimulation treatments for various neurological and psychiatric disorders, it appears crucial to consider both the opportunities and dangers such therapies may offer. In this review, we focus on the memory-modifying potential of optogenetics, investigating what it is capable of and how it differs from other memory modification technologies (MMTs). We then outline the safety challenges that need to be addressed before optogenetics can be used in humans. Finally, we re-examine crucial neuroethical concerns expressed in regard to other MMTs in the light of optogenetics and address those that appear to be unique to the memory-modifying potential of optogenetic technology. [ABSTRACT FROM AUTHOR]

Thermally selected male rainbow trout selectively bred at high temperatures were crossed with females of the Nikko strain with normal thermal tolerance. The F2 pedigrees, comprising 1641 individuals, were divided into 26 batches. The fish were acclimated to 17 °C, then their thermal tolerance determined by measuring the effective time (ET) required for them to lose equilibrium at an approximate upper lethal temperature (28 °C). The fish were categorized into 0- to < 20-, 20- to < 40-, 40- to < 60-, and ≤ 60-min ET groups. The individuals that survived the first treatment were subjected to a second experiment to determine the ET again for their respective groups. High reproducibility, i.e., 100%, for the 0- to < 20-min ET group was observed in the third experiment for individuals grouped into this ET category in the second experiment; 85.5% of individuals classified into the ≤ 60-min ET in the first experiment were recovered in the second experiment. Thus, our findings demonstrate that ET is a useful indicator for discriminating phenotypes according to their thermal tolerance. [ABSTRACT FROM AUTHOR]

Movement disorders are neurological conditions in which patients manifest a diverse range of movement impairments. Distinct structures within the basal ganglia of the brain, an area involved in movement regulation, are differentially affected for every disease. Among the most studied movement disorder conditions are Parkinson's (PD) and Huntington's disease (HD), in which the deregulation of the movement circuitry due to the loss of specific neuronal populations in basal ganglia is the underlying cause of motor symptoms. These symptoms are due to the loss principally of dopaminergic neurons of the substantia nigra (SN) par compacta and the GABAergic neurons of the striatum in PD and HD, respectively. Although these diseases were described in the 19th century, no effective treatment can slow down, reverse, or stop disease progression. Available pharmacological therapies have been focused on preventing or alleviating motor symptoms to improve the quality of life of patients, but these drugs are not able to mitigate the progressive neurodegeneration. Currently, considerable therapeutic advances have been achieved seeking a more efficacious and durable therapeutic effect. Here, we will focus on the new advances of several therapeutic approaches for PD and HD, starting with the available pharmacological treatments to alleviate the motor symptoms in both diseases. Then, we describe therapeutic strategies that aim to restore specific neuronal populations or their activity. Among the discussed strategies, the use of Neurotrophic factors (NTFs) and genetic approaches to prevent the neuronal loss in these diseases will be described. We will highlight strategies that have been evaluated in both Parkinson's and Huntington's patients, and also the ones with strong preclinical evidence. These current therapeutic techniques represent the most promising tools for the safe treatment of both diseases, specifically those aimed to avoid neuronal loss during disease progression. [ABSTRACT FROM AUTHOR]

Background: Ataxia with oculomotor apraxia type 1 (AOA1) is a rare autosomal recessive cerebellar ataxia, caused by mutations in the APTX gene. The disease is characterized by early-onset cerebellar ataxia, oculomotor apraxia and severe axonal polyneuropathy. The aim of this study was to detect the disease-causing variants in two unrelated consanguineous Jordanian families with cerebellar ataxia using whole exome sequencing (WES), and to correlate the identified mutation(s) with the clinical and cellular phenotypes. Methods: WES was performed in three affected individuals and segregation analysis of p.W279* APTX candidate variant was performed. Expression levels of APTX were measured in patients' skin fibroblasts and peripheral blood mononuclear cells, followed by western blot analysis in skin fibroblasts. Genotoxicity assay was performed to detect the sensitivity of APTX mutated cells to H2O2, MMC, MMS and etoposide. Results: A recurrent homozygous nonsense variant in APTX gene (c.837G>A, p.W279*) was revealed in all affected individuals. qRT-PCR showed normal APTX levels in peripheral blood and lower levels in fibroblast cells. However, western blot showed the absence of APTX protein in patients' skin fibroblasts. Significant hypersensitivity to H2O2, MMC and etoposide and lack of sensitivity to MMS were noted. Conclusions: This is the first study to report the identification of a nonsense variant in the APTX gene (c.837G>A; p.W279*) in AOA1 patients within the Jordanian population. This study confirmed the need of WES to assist in the diagnosis of cerebellar ataxia and it emphasizes the importance of studying the pathophysiology of the APTX gene. [ABSTRACT FROM AUTHOR]

Gene therapy (GT) has tremendous potential for the treatment of neurological disorders to transform patient care. The successful application of virus-mediated GT to treat spinal muscular atrophy is a significant milestone, serving to accelerate similar progress in a spectrum of neurological conditions, with more than 50 clinical trials currently underway, across neurodevelopmental, neurodegenerative, muscular dystrophy, epilepsy, chronic pain and neoplastic diseases. This review provides an overview of the key features of virus-mediated GT, paradigms of delivery and dosing, potential risks and highlights ongoing research to optimise safe and effective delivery of vectors into the nervous system. Examples of the application of GT in various neurological diseases alongside clinical development challenges will be presented. As the development and translation of GTs gain pace, success can only ultimately be realised for patients following implementation in the health system. The challenges and controversies of daunting costs, ethics, early diagnosis and health system readiness will require innovative pricing schemes, regulatory policies, education and organisation of a skilled workforce to deliver of high-quality care in clinical practice as we prepare for advanced therapeutics in neurology. [ABSTRACT FROM AUTHOR]

Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is clinically characterized by early-onset dementia, stroke, spondylosis deformans, and alopecia. In CARASIL cases, brain magnetic resonance imaging reveals severe white matter hyperintensities (WMHs), lacunar infarctions, and microbleeds. CARASIL is caused by a homozygous mutation in high-temperature requirement A serine peptidase 1 (HTRA1). Recently, it was reported that several heterozygous mutations in HTRA1 also cause cerebral small vessel disease (CSVD). Although patients with heterozygous HTRA1 -related CSVD (symptomatic carriers) are reported to have a milder form of CARASIL, little is known about the clinical and genetic differences between the two diseases. Given this gap in the literature, we collected clinical information on HTRA1 -related CSVD from a review of the literature to help clarify the differences between symptomatic carriers and CARASIL and the features of both diseases. Forty-six symptomatic carriers and 28 patients with CARASIL were investigated. Twenty-eight mutations in symptomatic carriers and 22 mutations in CARASIL were identified. Missense mutations in symptomatic carriers are more frequently identified in the linker or loop 3 (L3)/loop D (LD) domains, which are critical sites in activating protease activity. The ages at onset of neurological symptoms/signs were significantly higher in symptomatic carriers than in CARASIL, and the frequency of characteristic extraneurological findings and confluent WMHs were significantly higher in CARASIL than in symptomatic carriers. As previously reported, heterozygous HTRA1 -related CSVD has a milder clinical presentation of CARASIL. It seems that haploinsufficiency can cause CSVD among symptomatic carriers according to the several patients with heterozygous nonsense/frameshift mutations. However, the differing locations of mutations found in the two diseases indicate that distinct molecular mechanisms influence the development of CSVD in patients with HTRA1 -related CSVD. These findings further support continued careful examination of the pathogenicity of mutations located outside the linker or LD/L3 domain in symptomatic carriers. [ABSTRACT FROM AUTHOR]

Complementary DNA encoding a part of myosin heavy chain was cloned from fast skeletal muscle of Japanese codling Physiculus japonicus. Japanese codling, a frequent catch off the Pacific coast of northeastern Japan, is a typical underutilized fish species, especially in summer. Phylogenetic analysis of the deduced amino acid sequence of Japanese codling myosin heavy chain loop 2 region revealed that this fish belongs to a group comprising white croaker Pennahia argentata and walleye pollack Gadus chalcogrammus, which are commonly used as raw materials for surimi-based products. Thus Japanese codling meat was considered a promising candidate ingredient for surimi-based products. Salt-ground meat was prepared from Japanese codling in the presence of 0.5 M NaCl and subjected to a two-step heating procedure: pre-heating at 15–70 °C for 30 min, and secondary heating at 85 °C for 20 min. Thermal gels with a maximum breaking strength of > 10 N were produced by this process. The breaking strength was comparable to that of white croaker and walleye pollack. On the basis of the phylogenetic analysis of myosin heavy chains, and the properties of the heat-induced surimi gel, we conclude that Japanese codling is a promising raw material for the production of surimi-based products. [ABSTRACT FROM AUTHOR]

Electrochemical profiles of Pueraria bouffordii, P. montana var. lobata, P. montana var. montana, P. montana var. thomsonii, Haymondia wallichii, Teyleria stricta and Toxicopueraria yunnanensis were recorded from leaf tissue after different solvent extractions. The voltammetric data recorded after different solvent extractions can be derived as patterns for species identification. The electrochemical behavior of plant tissue contains its electrochemical active compounds profile. As the distribution of chemical compounds in plants is controlled by genes, these profiles can reflect differences at the genetic level between species. The dendrogram deduced from the electrochemical profile has been used for polyphyly analysis. The result suggests the Teyleria stricta showed very distant relationships with other species. P. montana var. lobata, P. montana var. montana and P. montana var. thomsonii showed a close relationship because they were varietas. Interestingly, H. wallichii showed a close infrageneric relationship within these species, which disagrees with other morphological studies. In addition, the result also provides insight into phylogenetic status of the regionally Toxicopueraria yunnanensis. [ABSTRACT FROM AUTHOR]