A systematic review of salivary biomarkers in Parkinson's disease

The search fo r reliable and easily accessible biomarkers in Parkinson's disease is receiving a growing emphasis,to detect neurodegeneration from the prodromal phase and to enforce disease-modifying therapies.Despite the need for non-invasively accessible biomarke rs,the majo rity of the studies have pointed to cerebrospinal fluid or peripheral biopsies biomarkers,which require invasive collection procedures.Saliva represents an easily accessible biofluid and an incredibly wide source of molecular biomarkers.In the present study,after presenting the morphological and biological bases for looking at saliva in the search of biomarkers for Parkinson's disease,we systematically reviewed the results achieved so far in the saliva of different cohorts of Parkinson's disease patients.A comprehensive literature search on PubMed and SCOPUS led to the discovery of 289articles.After screening and exclusion,34 relevant articles were derived fo r systematic review.Alpha-synuclein,the histopathological hallmark of Parkinson's disease,has been the most investigated Parkinson's disease biomarker in saliva,with oligomeric alphasynuclein consistently found increased in Parkinson's disease patients in comparison to healthy controls,while conflicting results have been reported regarding the levels of total alpha-synuclein and phosphorylated alpha-synuclein,and few studies described an increased oligomeric alpha-synuclein/total alpha-synuclein ratio in Parkinson's disease.Beyond alpha-synuclein,other biomarkers to rgeting diffe rent molecular pathways have been explored in the saliva of Parkinson's disease patients:total tau,phosphorylated tau,amyloid-β1-42(pathological protein aggregation biomarkers);DJ-1,heme-oxygenase-l,metabolites(alte red energy homeostasis biomarkers);MAPLC-3beta(aberrant proteostasis biomarker);cortisol,tumor necrosis factor-alpha(inflammation biomarkers);DNA methylation,miRNA(DNA/RNA defects biomarkers);acetylcholinesterase activity(synaptic and neuronal network dysfunction biomarkers);Raman spectra,proteome,and caffeine.Despite a few studies investigating biomarkers to rgeting molecular pathways different from alpha-synuclein in Parkinson's disease,these results should be replicated and observed in studies on larger cohorts,considering the potential role of these biomarkers in determining the molecular variance among Parkinson's disease subtypes.Although the need fo r standardization in sample collection and processing,salivary-based biomarkers studies have reported encouraging results,calling for large-scale longitudinal studies and multicentric assessments,given the great molecular potentials and the non-invasive accessibility of saliva.

Physical exercise and synaptic protection in human and pre-clinical models of multiple sclerosis

In multiple sclerosis,only immunomodulato ry and immunosuppressive drugs are recognized as disease-modifying therapies.Howeve r,in recent years,several data from pre-clinical and clinical studies suggested a possible role of physical exe rcise as disease-modifying therapy in multiple sclerosis.Current evidence is sparse and often conflicting,and the mechanisms underlying the neuroprotective and antinflammatory role of exercise in multiple sclerosis have not been fully elucidated.Data,mainly derived from pre-clinical studies,suggest that exe rcise could enhance longterm potentiation and thus neuroplasticity,could reduce neuroinflammation and synaptopathy,and dampen astrogliosis and microgliosis.In humans,most trials focused on direct clinical and MRI outcomes,as investigating synaptic,neuroinflammato ry,and pathological changes is not straightfo rward compared to animal models.The present review analyzed current evidence and limitations in research concerning the potential disease-modifying therapy effects of exercise in multiple sclerosis in animal models and human studies.

Far lateral lumbar disc herniation part 1: Imaging, neurophysiology and clinical features

Far lateral lumbar disc herniations(FLLDH)represent a separate category of disc pathology which includes both intraforaminal and extraforaminal lumbar disc herniations,that are characterized by a peculiar clinical presentation,diagnostic and treatment modalities as compared to the more frequent median and paramedian disc hernias.Surgical treatment often represents the only effective weapon for the cure of this disease and over the years different approaches have been developed that can reach the region of the foramen or external to it,with different degrees of invasiveness.The diagnosis is more demanding and still underestimated as it requires a more detailed knowledge in the spine anatomy and dedicated radiological studies.Computerized tomography and in particular magnetic resonance imaging are the appropriate tools for the diagnosis of FLLDH.Despite the widespread use of these diagnostic tests,many cases of FLLDH are overlooked due to insufficiently detailed radiological examinations or due to the execution of exams not focused to the foraminal or the extraforaminal region.Neurophysiological studies represent a valid aid in the diagnostic classification of this pathology and in some cases they can facilitate the differential diagnosis with other types of radiculopathies.In the present study,a comprehensive review of the clinical presentation,epidemiology,radiological study and the neurophysiological aspects is presented.

An eighteen-month follow-up study on the effects of Intravitreal Dexamethasone Implant in diabetic macular edema refractory to anti-VEGF therapy

AIM： To evaluate the long-term efficacy and safety of dexamethasone implants in subjects affected by diabetic macular edema（DME） resistant to anti-vascular endothelial growth factor（VEGF） therapy.METHODS： Thirty-two DME patients were enrolled.A700 microgram slow release Intravitreal Dexamethasone Implant（Ozurdex~） was placed in the vitreous cavity.All patients were followed for 18 mo.Best-corrected visual acuity（BCVA） measured with Early Treatment Diabetic Retinopathy Study（ETDRS） and central macular thickness（CMT） exams were carried out at baseline（T0）and after 1（T1）,3（T3）,4（T4）,6（T6）,9（T9）,12（T12）,15（T15）,and 18mo（T18） post injection. RESULTS： Repeated measures ANOVA showed an effect of treatment on ETDRS（P〈0.0001）.Post hoc analyses revealed that ETDRS values were significantly increased at T1,T3,T4,T9,and T15（P 〈0.001） as compared to baseline value（T0）.At T6,T12,and T18,ETDRS values were still statistically higher than baseline（P〈0.001 vs T0）.However,at these time points,we observed a trend to return to baseline conditions.ANOVA also showed an effect of treatment（P 〈0.0001）.CMT decreased significantly at T1,T3,T4,T9,and T15（P〈0.001）.At T6（P〈0.01）,T12 and T18（P〈0.001） CMT was also significantly lower than T0 although a trend to return to the baseline conditions was also observed.CONCLUSION： Our findings demonstrate that Intravitreal Dexamethasone Implant is a good option to improveBCVA and CMT in DME patients resistant to anti-VEGF therapy.Our data also show that the use of drugs administered directly into the vitreous allows achieving appropriate and long-lasting concentration at the site of disease without systemic side effects.

Mesenchymal stem cells in neurodegenerative diseases: Opinion review on ethical dilemmas

The treatment of neurodegenerative diseases presents a growing need for innovation in relation to recent evidence in the field of reconstructive therapy using stem cells. Understanding the molecular mechanisms underlying neurodegenerative disorders, and the advent of methods able to induce neuronal stem cell differentiation allowed to develop innovative therapeutic approaches offering the prospect of healthy and perfectly functional cell transplants, able to replace the sick ones. Hence the importance of deepening the state of the art regarding the clinical applications of advanced cell therapy products for the regeneration of nerve tissue. Besides representing a promising area of tissue transplant surgery and a great achievement in the field of neurodegenerative disease, stem cell research presents certain critical issues that need to be carefully examined from the ethical perspective. In fact, a subject so complex and not entirely explored requires a detailed scientific and ethical evaluation aimed at avoiding improper and ineffective use, rather than incorrect indications, technical inadequacies, and incongruous expectations. In fact, the clinical usefulness of stem cells will only be certain if able to provide the patient with safe, long-term and substantially more effective strategies than any other treatment available.The present paper provides an ethical assessment of tissue regeneration through mesenchymal stem cells in neurodegenerative diseases with the aim to rule out the fundamental issues related to research and clinical translation.

A mutation in the ZNF687 gene that is responsible for the severe form of Paget's disease of bone causes severely altered bone remodeling and promotes hepatocellular carcinoma onset in a knock-in mouse model

Paget’s disease(PDB)is a late-onset bone remodeling disorder with a broad spectrum of symptoms and complications.One of the most aggressive forms is caused by the P937R mutation in the ZNF687 gene.Although the genetic involvement of ZNF687 in PDB has been extensively studied,the molecular mechanisms underlying this association remain unclear.Here,we describe the first Zfp687 knock-in mouse model and demonstrate that the mutation recapitulates the PDB phenotype,resulting in severely altered bone remodeling.Through microcomputed tomography analysis,we observed that 8-month-old mutant mice showed a mainly osteolytic phase,with a significant decrease in the trabecular bone volume affecting the femurs and the vertebrae.Conversely,osteoblast activity was deregulated,producing disorganized bone.Notably,this phenotype became pervasive in 16-month-old mice,where osteoblast function overtook bone resorption,as highlighted by the presence of woven bone in histological analyses,consistent with the PDB phenotype.Furthermore,we detected osteophytes and intervertebral disc degeneration,outlining for the first time the link between osteoarthritis and PDB in a PDB mouse model.RNA sequencing of wild-type and Zfp687 knockout RAW264.7 cells identified a set of genes involved in osteoclastogenesis potentially regulated by Zfp687,e.g.,Tspan7,Cpe,Vegfc,and Ggt1,confirming its role in this process.Strikingly,in this mouse model,the mutation was also associated with a high penetrance of hepatocellular carcinomas.Thus,this study established an essential role of Zfp687 in the regulation of bone remodeling,offering the potential to therapeutically treat PDB,and underlines the oncogenic potential of ZNF687.

Percutaneous coronary intervention in nonagenarians: pros and cons

Percutaneous coronary intervention is a mainstay in the management of symptomatic or high-risk coronary artery disease. The bulk of clinical evidence and experience underlying this fact relies, however, on relatively young patients. Indeed, few data of very limited quality are available which adequately define the risk-benefit and cost-benefit profile of coronary angioplasty and stenting in very old subjects, such as those of 90 years of age or older (i.e., nonagenarians). The aim of this review is to provide a concise, yet practical, synthesis of the available evidence on percutaneous coronary revascularization in the very elderly. The main arguments elaborated upon are to what extent we can extrapolate findings from studies including younger patients to nonagenarians, whether we should provide higher priority to prognosis or quality of life in such patients, and whether we can afford to allocate vast resources to care for such subjects in an era of financial constraints. Our review of 18 studies and 1082 patients suggest that percutaneous coronary intervention is feasible and associated with acceptable short- and long-term results in this population, which is nonetheless fraught with a high mortality risk irrespective of the revascularization procedure. Accordingly, the pros and cons of percutaneous coronary intervention should be carefully weighed when considering this treatment in nonagenarians.

Glyco-sphingo biology: a novel perspective for potential new treatments in Huntington's disease

Huntington＇s disease （HD） is the most common dominantly inherited neurodegenerative disorder, mainly characterized by the progressive striatal and cortical neurodegeneration and as- sociated motor, cognitive and behavioural disturbances （Zuccato et al., 2010）. The disease-causing mutation is an expansion of a GAG trinucleotide repeat （〉 36 repeats） encoding a polygluta- mine stretch in the N-terminal region ofhuntingtin （Htt） （Zuc- cato et al., 2010）, a ubiquitous protein whose function is still unclear （Zuccato et al., 2010）. Expansion of the polyQ stretch endows mutant Htt （mHtt） with toxic properties, and results in the development of a broad array of undesirable effects in both neuronal and non-neuronal cells （Zuccato et al., 2010）. Among all cellular dysfunctions and biochemical imbalances classically associated with HD, perturbed metabolism of （glyco） sphingolipids appears to play a crucial role in the pathogenesis of the disease. Over the last years, we and other have extensively contributed to these findings （Desplats et al., 2007;

Detailing the ultrastructure’s increase of prion protein in pancreatic adenocarcinoma

BACKGROUND Recent evidences have shown a relationship between prion protein(PrPc)expression and pancreatic ductal adenocarcinoma(PDAC).Indeed,PrPc could be one of the markers explaining the aggressiveness of this tumor.However,studies investigating the specific compartmentalization of increased PrPc expression within PDAC cells are lacking,as well as a correlation between ultrastructural evidence,ultrastructural morphometry of PrPc protein and clinical data.These data,as well as the quantitative stoichiometry of this protein detected by immuno-gold,provide a significant advancement in understanding the biology of disease and the outcome of surgical resection.AIM To analyze quantitative stoichiometry and compartmentalization of PrPc in PDAC cells and to correlate its presence with prognostic data METHODS Between June 2018 and December 2020,samples from pancreatic tissues of 45 patients treated with pancreatic resection for a preoperative suspicion of PDAC at our Institution were collected.When the frozen section excluded a PDAC diagnosis,or the nodules were too small for adequate sampling,patients were ruled out from the present study.Western blotting was used to detect,quantify and compare the expression of PrPc in PDAC and control tissues,such as those of non-affected neighboring pancreatic tissue of the same patient.To quantify the increase of PrPc and to detect the subcellular compartmentalization of PrPc within PDAC cells,immuno-gold stoichiometry within specific cell compartments was analyzed with electron microscopy.Finally,an analysis of quantitative PrPc expression according to prognostic data,such as cancer stage,recurrence of the disease at 12 mo after surgery and recurrence during adjuvant chemotherapy was made.RESULTS The amount of PrPc within specimen from 38 out of 45 patients was determined by semi-quantitative analysis by using Western blotting,which indicates that PrPc increases almost three-fold in tumor pancreatic tissue compared with healthy pancreatic regions[242.41±28.36 optical density(OD)vs 95±17.40 OD,P<0.0001].Quantitative morphometry carried out by using immuno-gold detection at transmission electron microscopy confirms an increased PrPc expression in PDAC ductal cells of all patients and allows to detect a specific compartmentalization of PrPc within tumor cells.In particular,the number of immuno-gold particles of PrPc was significantly higher in PDAC cells respect to controls,when considering the whole cell(19.8±0.79 particles vs 9.44±0.45,P<0.0001).Remarkably,considering PDAC cells,the increase of PrPc was higher in the nucleus than cytosol of tumor cells,which indicates a shift in PrPc compartmentalization within tumor cells.In fact,the increase of immuno-gold within nuclear compartment exceeds at large the augment of PrPc which was detected in the cytosol(nucleus:12.88±0.59 particles vs 5.12±0.32,P<0.0001;cytosol:7.74.±0.44 particles vs 4.3±0.24,P<0.0001).RESULTS In order to analyze the prognostic impact of PrPc,we found a correlation between PrPc expression and cancer stage according to pathology results,with a significantly higher expression of PrPc for advanced stages.Moreover,24 patients with a mean follow-up of 16.8 mo were considered.Immuno-blot analysis revealed a significantly higher expression of PrPc in patients with disease recurrence at 12 mo after radical surgery(360.71±69.01 OD vs 170.23±23.06 OD,P=0.023),also in the subgroup of patients treated with adjuvant CT(368.36±79.26 OD in the recurrence group vs 162.86±24.16 OD,P=0.028),which indicates a correlation with a higher chemo-resistance.CONCLUSION Expression of PrPc is significantly higher in PDAC cells compared with control,with the protein mainly placed in the nucleus.Preliminary clinical data confirm the correlation with a poorer prognosis.

Comparative Oral Absorption of Different Citicoline and Homotaurine Formulations: A Single-Dose, Two-Period Crossover Trial in the Dog

Background: Citicoline and homotaurine are compounds with a potent neuroprotective activity and they have been administered for many years in the treatment of numerous neurodegenerative and ophthalmological diseases, including glaucoma. Initially available only as liquid form, through parenteral route, nowadays citicoline can be administered also as tablet but no data on bioavailability of these different forms are available. In the present study, pharmacokinetics of citicoline in tablet versus vials, each at the therapeutic dose of 500 mg, in addition to 50 mg of homotaurine was investigated. Materials and methods: Ten mixed breed dogs received a single dose of 50 mg oral homotaurine and 500 mg citicoline in tablet and vials with the same dose were administered after a seven days wash-out period. Parameters assessed for citicoline metabolites (cytidine, uridine and choline) were AUC0&#8722;t, Cmax and Tmax. Results: Citicoline bioavailability appeared to be slightly higher for the tablet compared to the vial formulation. Cytidine is equivalent in absorption dynamics both for tablet and liquid form;uridine for tablet reaches its maximum and is reabsorbed more quickly while choline for the liquid form reaches the maximum first and is reabsorbed more quickly. Conclusions: Citicoline in tablet and liquid formulation have pharmacokinetic properties leading to a very similar bioavailability.

γ氨基丁酸转运体1在大脑皮质小胶质细胞中的表达

小胶质细胞是脑的免疫细胞。通过感知微环境,小胶质细胞可以使大脑正常发育和发挥功能。它们与其他神经胶质细胞和神经元通讯,释放能对周围细胞产生影响的许多分子,并且能对这些分子做出反应。其中,神经递质,尤其是γ-氨基丁酸(GABA)最近引起了研究者对这一过程的兴趣。我们证明了GABA转运体1(GAT-1)在小胶质细胞的胞体和突起均有表达。我们发现,用强效的和选择性的GAT-1抑制剂NNC-711处理小胶质细胞,可显着降低其钠离子依赖性GABA的摄取。另一方面,用GAT-2/3抑制剂SNAP-5114处理细胞能显著增加其对GABA的摄取。该作用可被肉毒杆菌毒素Bo NT/C1完全阻断,该毒素特异性切割和灭活突触融合蛋白1A(STX1A)。总而言之,这些发现表明小胶质细胞表达GAT-1,且STX1A在小胶质细胞中GAT-1依赖的GABA摄取调控中起着重要作用。

小胶质细胞控制成年小鼠海马中的谷氨酸能突触

小胶质细胞是调节大脑突触发育和可塑性的重要细胞类型,但其影响突触的正常功能的机制尚不清楚。在本研究中,我们通过PLX5622造成小胶质细胞耗竭,并观察其对成年野生型小鼠海马CA3-CA1突触的影响。在小胶质细胞耗竭后,与树突棘密度降低相关的自发和诱发谷氨酸能活动的减少,出现未成熟突触特征以及突触的可塑性提高。小胶质细胞耗竭的小鼠在新物体识别任务的获取方面表现出缺陷。海马星形胶质细胞出现增生,但并没有神经炎症反应。在Cx3cr1-/-小鼠中,PLX不能导致海马出现上述改变。这说明CX3CL1/CX3CR1轴在小胶质细胞对突触功能的控制中有重要作用。PLX5622停用后,小胶质细胞的重新增殖,海马突触恢复,小鼠的学习功能也出现恢复。综上所述,小胶质细胞对维持成人大脑的突触的正常功能用重要的作用,去除小胶质细胞会导致谷氨酸能突触组织和活动的可逆变化。

通过EC2~黏附放置头皮电极在长时间视频EEG监护中的作用

Objective: To evaluate the usefulness of an adhesive paste named EC2(Grass-Telefactor) in comparison with collodion, for scalp electrodes placement in patients with drug resistant partial epilepsy monitored by long-term video-EEG. Methods: A total of 40 patients with drug resistant partial epilepsy participated in the study. In 20 patients, electrode placement on the scalp was made with collodion (group C)-whereas in the remaining patients EC2was used (Group P). After the electrode placement (T1) and after 24 h of recording (T2), the impedance of the electrodes was measured. Moreover, the time required to apply the electrodes and for their daily maintenance was calculated and recorded for all patients who entered the study. Results: At each observation, group C showed mean values of electrode impedance significantly higher that the group P (T1: 16.8 kΩ; T2: 6.5 kΩvs T1: 2.4 kΩ; T2: 4.0 kΩ, respectively) (P < 1×10-5). The time required to make the montage and to provide its daily maintenance was significantly shorter in group P than in group C [20.8 and 10.5 min vs 44.3 and 19.7 min, respectively (P < 1×10-5)]. Conclusions: We found that the use of EC2 paste in scalp electrode attachment is less time consuming, with better recording quality as a result of lower electrode impedance values, than the use of collodion. Significance: EC2 paste may substitute collodion in electrode placement for long-term video-EEG monitoring, with an optimal cost-benefit ratio in terms of recording performance, time consumption, and safety.

促纤维增生／结节型髓母细胞瘤病理学特点及相关信号通路研究

目的探讨促纤维增生／结节型髓母细胞瘤（DNMB）的组织病理学特征和瘤细胞分化相关的信号通路。方法采用HE染色、Reticulin染色和免疫组织化学染色等方法，对803例髓母细胞瘤石蜡样本进行分析。结果103例样本（103／803）确诊为DNMB，HE染色显示DNMB存在无网状纤维的结节样苍白岛，并由致密的多形性细胞及网状纤维围绕。所有样本Reticulin染色后，可见结节间存在银染成黑色的网状纤维。免疫组化结果：结节内细胞呈现胞质NSE、Syn阳性表达，胞核Neu-N阳性表达；结节内Ki-67标记指数与结节间相比明显较低；GABl和filaminA抗体染色显示结节间及周围细胞胞质阳性，结节内细胞部分表达；B-catenin抗体染色显示结节内、外细胞胞质阳性，但胞核均为阴性。结论DNMB是髓母细胞瘤中组织学形态较为特殊的亚型，Reticulin染色是重要的诊断方法，该肿瘤细胞分化与SHh信号通路激活存在密切联系。

Childhood absence epilepsy and benign epilepsy with centro-temporal spikes: a narrative review analysis

Background:Recent studies have shown a possible coexistence of absence seizures with other forms of epilepsy.The purpose of this study was to ascertain the possible contemporary or subsequent presence of childhood absence epilepsy (CAE) and benign epilepsy with centro-temporal spikes (BECTS) in pediatric epileptic patients.Data sources:A PubMed systematic search indexed for MEDLINE,PubMed and EMBASE was undertaken to identify studies in children including articles written between 1996 and 2015.Retrospective studies,meta-analysis and case reports were included.The list of references of all the relevant articles was also studied.The date of our last search was December 2015.Results:Review of the literature revealed 19 cases,8 females and 11 males,reporting a consecutive or contemporary coexistence of CAE and BECTS within the same patients.Patient's age ranged between 4 and 12 years.Three out of 19 patients presented concomitant features of both syndromes,whereas 16 patients experienced the two syndromes at different times.Conclusions:BECTS and CAE may be pathophysiologically related,and the two epileptic phenotypes may indicate a neurobiological continuum.Further studies are needed to elucidate a probable genetic or functional link between partial and primarily generalized electro-clinical patterns in idiopathic childhood epilepsies.

CUBE, a Cable-driven Device for Limb Rehabilitation

In this paper, a novel cable-driven parallel robot, CUBE, is introduced for the assistance of patients in rehabilitation exercising of both upper and lower limbs. The system is characterized by a lightweight structure that is easy to set-up and operate, for both clinical and home usage for both pre-determined and customized exercises, with control over the position of the end-effector while locking its rotation around the horizontal axes. Its cable-driven design makes it inherently safe in human/robot interactions also due to the extremely low inertia. While a novel end-effector design makes the device wearable both on the upper and lower limbs without having to disassemble any part of the structure. The design is presented with its kinematic analysis. Then, the manufacturing through 3D?printing and commercial components of a first prototype is reported. Finally, the system is validated through motion tests along simple trajectories and two different spatial exercises.