Haptoglobin Modulates Beta-Amyloid Uptake by U-87 MG Astrocyte Cell Line

Accumulation of beta-amyloid (Aβ) in the extracellular space, which is one of the hallmarks of Alzheimer’s disease (AD), depends on the balance between its synthesis and clearance. The physiological role of extracellular chaperones, capable of affecting early events in the amyloid cascade, is increasingly being investigated by many research groups. Among these proteins, we focused on haptoglobin, which we recently found to form a complex with beta-amyloid in brain tissues or cerebrospinal fluids from patients with AD.We also previously reported that haptoglobin increases with age in rat hippocampus. Major aimof this study was to evaluate whether haptoglobin influences Aβ interaction with astrocytes and its internalization into these cells. Haptoglobin effect on Aβ- induced cell death was also explored. We report here that haptoglobin impairs Aβ uptake by human glioblastoma–astrocytoma cell line U-87 MG and limits the toxicity of this peptide on these cells. Of note, our data also show that Aβ can stimulate haptoglobin release by astrocyte cell lines. The study of the risk of developing AD should be focused not only on the analysis ofAβ but also on the level of critical ligands, such as haptoglobin, able to influence peptide aggregation or clearance

Effect of Lupinus albus as protein supplement on yield, constituents, clotting properties and fatty acid composition in ewes' milk

The effect of feeding lupin seeds (Lupinus albus L.) as an alternative protein source in ewe diets was investigated. Two groups of 18 Sarda ewes were fed two different isonitrogenous diets: with lupin (L) seed, given after 12 h soaking, or soybean meal (SBM) as the main protein source. DMI, variations of body weight and milk production were unaffected by the treatment. Although not statistically significant, in the group fed L diet the production of milk fat and protein was higher. Clotting properties of milk were similar for the two treatments, probably due to the small differences in the milk protein contents. The fatty acid profile of milk was affected by treatment with a larger content of short (14.19 wt% versus 12.26 wt%)- and medium (49.37 wt% versus 47.76 wt%)-chain fatty acids in milk from ewes fed the L diet. CLA content was unaffected by treatment. Triglyceride content of fat from the two diets reflects the milk fatty acid composition. Indeed, milk from L diet showed a higher level of medium-chain triglycerides, which are of particular interest to consumers with concerns over health and heart disease. The inclusion of lupin seed in the diet of lactation ewes can be a means of achieving a more desirable triglyceride profile in milk fat. Milk with enhanced nutritive quality may promote wider market penetration of sheep dairy products

A Novel null homozygous mutation confirms <i>CACNA2D2</i> as a gene mutated in epileptic encephalopathy

Contribution to epileptic encephalopathy (EE) of mutations in CACNA2D2, encoding α2δ-2 subunit of Voltage Dependent Calcium Channels, is unclear. To date only one CACNA2D2 mutation altering channel functionality has been identified in a single family. In the same family, a rare CELSR3 polymorphism also segregated with disease. Involvement of CACNA2D2 in EE is therefore not confirmed, while that of CELSR3 is questionable. In a patient with epilepsy, dyskinesia, cerebellar atrophy, psychomotor delay and dysmorphic features, offspring to consanguineous parents, we performed whole exome sequencing (WES) for homozygosity mapping and mutation detection. WES identified extended autozygosity on chromosome 3, containing two novel homozygous candidate mutations: c.1295delA (p.Asn432fs) in CACNA2D2 and c.G6407A (p.Gly2136Asp) in CELSR3. Gene prioritization pointed to CACNA2D2 as the most prominent candidate gene. The WES finding in CACNA2D2 resulted to be statistically significant (p = 0.032), unlike that in CELSR3. CACNA2D2 homozygous c.1295delA essentially abolished α2δ-2 expression. In summary, we identified a novel null CACNA2D2 mutation associated to a clinical phenotype strikingly similar to the Cacna2d2 null mouse model. Molecular and statistical analyses together argued in favor of a causal contribution of CACNA2D2 mutations to EE, while suggested that finding in CELSR3, although potentially damaging, is likely incidental

Study of the effects of High Hydrostatic Pressure (HHP) and Pulsed Light (PL) on BSA structure and hydrolysis

Non-thermal technologies, such as High Hydrostatic Pressure (HHP) and Pulsed light (PL), affect protein inducing modifications in its conformational structure. For this reason the hydrolysis reaction of the protein can be modulated if it is conducted in combination with these technologies which are able to change the availability of peptide bonds exposed to the enzymatic action. The aim is to study the effects of HHP and PL on the structure and the extent of hydrolysis reaction of a target protein: the Bovine Serum Albumin (BSA). BSA protein (5 mg/mL) in sodium phosphate buffer (50 mM, pH = 8) were treated with PL and HHP at different processing conditions, namely pressure level and treatment time in the case of HHP and treatment time and energy input in the case of PL. Structural modification of the protein solutions were analyzed by determining the sulphidrilic groups and the changes of the secondary structure. The effect of the two treatments on the hydrolysis degree (HD) at 37 °C was also evaluated by OPA method. Chymotrypsin and trypsin (E/S ratio = 1/10) were used to hydrolyze the BSA protein solutions. The hydrolysis was carried out in HHP assisted or PL assisted conditions or the protein solutions were treated with HHP or PL processes and immediately after hydrolyzed with the enzymes. Results obtained so far demonstrated that the two technologies tested are able to induce protein modifications and the occurrence and importance of this phenomenon depends on processing parameters causing protein unfolding, namely pressure level and number of pulses. When the maximum protein unfolding is obtained, higher HD values are detected. The highest HD value is obtained in HHP assisted hydrolysis with longer treatment time, and when, before undergoing hydrolysis, the PL treatment is applied to the solution placed at the higher distance from the lamp

Role of Metal Ions and Hydrogen Bond Acceptors in the Tautomeric Equilibrium of Nitro-9[(Alkylamino)Amino]-Acridine Drugs

3-nitro-9-[2-(dialkylamino)ethyl)]aminoacridines (alkyl = methyl or ethyl) have been used as ligands towards platinum(If). The end product is a complex in which the acridine acts as a tridentate ligand contributing the two exocyclic nitrogen atoms and one of the two peri carbons. The metallation takes place predominantly at the peri position of the unsubstituted ring. The coordinated acridine is in the imino tautomeric form although, in the free state, it occurs exclusively in the amino form (both in the solid state and in solution). The imino tautomer is considered to be the biologically active form. In the platinated species the N(10)H of the acridine can be involved in strong hydrogen bonding with a chloride ion leading to formation of an association complex, the formation constant has been found to be 1.4±103 M−1. The N(10)H┄CI interaction can influence the tautomeric equilibrium of the acridine dye also in the uncoordinated species, however, the shift in favor of the imino tautomer is not complete

Isolated office hypertension : A 3-year follow-up study

The study aimed to evaluate, over a 3-year period, the progression towards sustained hypertension and left ventricular (LV) changes in patients with isolated office (IO) hypertension (office BP140 and/or 90 mmHg, daytime BP130/80 mmHg). After 3 years from the basal evaluation, 38 subjects with basal normal BP and 42 subjects with basal IO hypertension underwent a second 24-h BP monitoring and echocardiography; 19 patients of the basal IO hypertension group were not revaluated because they had already developed ambulatory hypertension and were on antihypertensive treatment. At the second evaluation, the 38 normotensive subjects had unchanged BP and LV parameters; 25 IO hypertensives have developed sustained hypertension. Considering them together with the 19 patients already treated, 72% of 61 IO hypertensives developed ambulatory hypertension over a 3-year period. The patients who subsequently developed hypertension differed from the group who did not only for lower basal values of LV diastolic parameters; all the patients with basal LV hypertrophy and/or preclinical diastolic impairment subsequently developed sustained hypertension. In conclusion, IO hypertensive patients show a high rate of progression towards sustained hypertension. Basal LV hypertrophy and/or preclinical diastolic dysfunction were the only markers of a greater risk of becoming hypertensives

Exercise stress testing in clinical practice

Exercise stress testing is an important diagnostic tool for evaluating patient’s cardiovascular performance. The present review describes the accuracy and the value of exercise stress testing in different settings: after an acute coronary event, after percutaneous coronary intervention or coronary artery bypass graft; in patients risk assessment before non-cardiac surgery; in diabetic population; in patients with baseline electrocardiographic abnormalities. Moreover, this review provides insights relating to test accuracy in women and geriatric patients. Finally, this review explores new variables/parameters (dyspnea, chronotropic incompentence, heart rate recovery, functional capacity, integrated scores) that in the last few years added an incremental value to conventional analysis of exercise-induced angina or electrocardiographic changes

A web-based tumor registration system for a regional Canine Cancer Registry in Umbria, central Italy

The Umbria Canine Cancer Registry (CCR) is a web-based platform for cancer registrationset up in order to estimate the incidence of spontaneous tumors. It is an integralpart of the regional canine demographic registry in which veterinary practitioners andpathologists interact. Veterinary pathologists perform double-blind comparisons andclassify neoplasms in an automated classification process using the WHO criteria forcanine neoplasms and the ICD-O tumor topographical and morphological keys. Herewe describe the organization, on-line procedures and the methods used to assess caninedemography, a pre-requisite for accurately estimating the incidence of cancer.In its first 4 years the CCR recruited 4857 cases of suspected tumors, as diagnosed bypractitioners, clinics and a veterinary hospital. After the first year the number of enrolledcases increased by 63%, suggesting growing interest from the regional veterinarycommunity

Anti-neuropathic effects of Rosmarinus officinalis L. terpenoid fraction: Relevance of nicotinic receptors

Traditional uses and current results highlight the neuroprotective properties of Rosmarinus officinalis L. The compelling need for novel strategies able to relieve neuropathic pain encouraged us to analyze different rosemary leaf extracts in rats following chronic constriction injury (CCI) of sciatic nerve. Ethanol, acetone, and the innovative ultrasound-hexane extractive methods were used to obtain: EE, AE, and for hexane extracts UREprel and URE. Extracts were characterized in terms of typical constituents and repeatedly administered to CCI-rats (13-days treatment, from the day of surgery). URE showed the best efficacy and potency in reducing hypersensitivity to noxious- and non-noxious stimuli and spontaneous pain. URE contained the higher quantity of the terpenoid carnosic acid (CA) and its efficacy was compared to pure CA. Histological analysis of the sciatic nerve revealed that URE prevented axon and myelin derangement, edema and inflammatory infiltrate. In the dorsal horn of the spinal cord, URE did not reduce astrocyte activation. Both the pain reliever and the neuroconservative effects of URE were significantly prevented by the nicotinic receptor (nAChR) antagonist mecamylamine. In conclusion, the hexane-ultrasound rosemary extract is able to reduce neuropathic hypersensitivity and protect nervous tissues. Effectiveness is mainly related to the terpenoid fraction by mechanisms involving nAChRs