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Outer membrane vesicles (OMVs) are nanostructures mostly produced by blebbing of the outer membrane in Gram negative bacteria. They contain biologically active proteins and perform a variety of processes. OMV production is also a typical response to events inducing stress in the bacterial envelope. In these cases, hypervesiculation is regarded as a strategy to avoid the dangerous accumulation of undesired products within the periplasm. Several housekeeping genes influence the biogenesis of OMVs, including those correlated with peptidoglycan and cell wall dynamics. In this work, we have investigated the relationship between OMV production and the lysis module of the E. coli DLP12 cryptic prophage. This module is an operon encoding a holin, an endolysin and two spannins, and is known to be involved in cell wall maintenance. We find that deleting the lysis module increases OMV production, suggesting that during evolution this operon has been domesticated to regulate vesiculation, likely through the elimination of non-recyclable peptidoglycan fragments. We also show that the expression of the lysis module is negatively regulated by environmental stress stimuli as high osmolarity, low pH and low temperature. Our data further highlight how defective prophages finely contribute to bacterial host fitness.

We recently demonstrated that HepaRG cells encapsulated into 1.5% alginate beads are capable of self-assembling into spheroids. They adequately differentiate into hepatocyte-like cells, with hepatic features observed at Day 14 post-encapsulation required for external bioartificial liver applications. Preliminary investigations performed within a bioreactor under shear stress conditions and using a culture medium mimicking acute liver failure (ALF) highlighted the need to reinforce beads with a polymer coating. We demonstrated in a first step that a poly-l-lysine coating improved the mechanical stability, without altering the metabolic activities necessary for bioartificial liver applications (such as ammonia and lactate elimination). In a second step, we tested the optimized biomass in a newly designed perfused dynamic bioreactor, in the presence of the medium model for pathological plasma for 6 h. Performances of the biomass were enhanced as compared to the steady configuration, demonstrating its efficacy in decreasing the typical toxins of ALF. This type of bioreactor is easy to scale up as it relies on the number of micro-encapsulated cells, and could provide an adequate hepatic biomass for liver supply. Its design allows it to be integrated into a hybrid artificial/bioartificial liver setup for further clinical studies regarding its impact on ALF animal models., (© 2020 Wiley Periodicals LLC.)

Efflux pumps (EPs) are present in all living cells and represent a large and important group of transmembrane proteins involved in transport processes. In bacteria, multidrug resistance efflux pumps (MDR EPs) confer resistance to antibiotics at different levels and are deeply implicated in the fast and dramatic emergence of antibiotic resistance. Recently, several reports have outlined the great versatility of MDR EPs in exporting a large variety of compounds other than antibiotics, thus promoting bacterial adaptation to a wide range of habitats. In several bacterial pathogens, MDR EPs contribute to increase the virulence potential and are directly involved in the crosstalk with host cells. In this work, we have investigated the possible role of MDR EPs in the infectious process of the adherent-invasive Escherichia coli (AIEC), a group of pathogenic E. coli that colonize the ileal mucosa of Crohn disease (CD) patients causing a strong intestinal inflammation. The results we have obtained indicate that, with the exception of mdtM , all MDR-EPs encoding genes present in E.coli K12 are conserved in the AIEC prototype strain LF82. The analysis of MDR EP expression during LF82 infection of macrophages and epithelial cells reveals that their transcription is highly modulated during the bacterial intracellular life. Notably, some EP genes are regulated in a cell-type specific manner, strongly suggesting that their function is required for LF82 successful infection. AIEC are able to adhere to and invade intestinal epithelial cells and, importantly, to survive and multiply within macrophages. Thus, we further investigated the role of EPs specifically induced by macrophage environment. We present evidence indicating that deletion of mdtEF genes, encoding an MDR EP belonging to the resistance nodulation division (RND) family, significantly impairs survival of LF82 in macrophages and that the wild type phenotype can be restored by trans-complementation with functional MdtEF pump. Altogether, our results indicate a strong involvement of MDR EPs in host pathogen interaction also in AIEC and highlight the contribution of MdtEF to the fitness of LF82 in the macrophage environment., (Copyright © 2020 Fanelli, Pasqua, Colonna, Prosseda and Grossi.)

In liver tissue engineering, cell culture in spheroids is now well recognized to promote the maintenance of hepatic functions. However, the process leading to spheroids formation is time consuming, costly, and not easy to scale-up for further use in human bioartificial liver (BAL) applications. In this study, we encapsulated HepaRG cells (precursors of hepatocyte-like cells) in 1.5% alginate beads without preforming spheroids. Starting from a given hepatic biomass, we analyzed cell differentiation and metabolic performance for further use in a fluidized-bed BAL. We observed that cells self-rearranged as aggregates within the beads and adequately differentiated over time, in the absence of any differentiating factors classically used. On day 14 postencapsulation, cells displayed a wide range of hepatic features necessary for the treatment of a patient in acute liver failure. These activities include albumin synthesis, ammonia and lactate detoxification, and the efficacy of the enzymes involved in the xenobiotic metabolism (such as CYP1A1/2). Impact statement It has been recognized that culturing cells in spheroids (SPHs) is advantageous as they better reproduce the three-dimensional physiological microenvironment. This approach can be exploited in bioartificial liver applications, where obtaining a functional hepatic biomass is the major challenge. Our study describes an original method for culturing hepatic cells in alginate beads that makes possible the autonomous formation of SPHs after 3 days of culture. In turn, the cells differentiate adequately and display a wide range of hepatic features. They are also capable of treating a pathological plasma model. Finally, this setup can easily be scaled-up to treat acute liver failure.

Efflux pumps represent an important and large group of transporter proteins found in all organisms. The importance of efflux pumps resides in their ability to extrude a wide range of antibiotics, resulting in the emergence of multidrug resistance in many bacteria. Besides antibiotics, multidrug efflux pumps can also extrude a large variety of compounds: Bacterial metabolites, plant-produced compounds, quorum-sensing molecules, and virulence factors. This versatility makes efflux pumps relevant players in interactions not only with other bacteria, but also with plant or animal cells. The multidrug efflux pumps belonging to the major facilitator superfamily (MFS) are widely distributed in microbial genomes and exhibit a large spectrum of substrate specificities. Multidrug MFS efflux pumps are present either as single-component transporters or as tripartite complexes. In this review, we will summarize how the multidrug MFS efflux pumps contribute to the interplay between bacteria and targeted host cells, with emphasis on their role in bacterial virulence, in the colonization of plant and animal host cells and in biofilm formation. We will also address the complexity of these interactions in the light of the underlying regulatory networks required for the effective activation of efflux pump genes., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Patients with allergic rhinitis (AR) can suffer from mood disorders. The aim of this study was to investigate the clinical effect of a liposomal nasal spray (LN) containing vitamins A and E on the nasal mucosa in patients suffering from AR who had refused any type of anti-allergic treatment. For this purpose, the results of nasal cytology, Visual Analog Scale (VAS), Sino-Nasal Outcome Test-22 (SNOT-22), and Hospital Anxiety and Depression Scale (HADS) test were analyzed. Moreover, we evaluated the relationship between SNOT-22 and nasal cytology and between nasal symptoms and HADS scores. Statistical analysis revealed a significant decrease of scores at T1 in the LN treatment group as concerns VAS, SNOT-22, HADS-Anxiety test and a remarkable reduction of inflammatory cells detected with nasal cytology. Our study showed that higher levels of SNOT-22 corresponded to a higher level of HADS-Anxiety. The mechanisms underlying this relationship in AR patients are currently unknown, but we can suppose that improving mucosal trophism may contribute to the decrease of nasal symptoms and anxiety scores. The improvement of nasal symptoms, as measured by SNOT-22, was significantly correlated with the objective results of nasal cytology. These relationships between SNOT-22 and nasal cytology and between anxiety and cytology were investigated for the first time in our research.

Elastin-like polypeptides (ELPs) are biocompatible-engineered polypeptides, with promising interest in tissue engineering due to their intrinsic biological and physical properties, and their ease of production. The IKVAV (Ile-Lys-Val-Ala-Val) laminin-1 sequence has been shown to sustain neuron attachment and growth. In this study, the IKVAV adhesion sequence, or a scrambled VKAIV sequence, were incorporated by genetic engineering in the structure of an ELP, expressed in Escherichia coli and purified. The transition temperatures of the ELP-IKVAV and ELP-VKAIV were determined to be 23 °C. Although the phase transition was fully reversible for ELP-VKAIV, we observed an irreversible aggregation for ELP-IKVAV. The corresponding aggregates shared some characteristics with amyloid-like polypeptides. The two ELPs were then reacted with functionalized polyethylene glycol (PEG) to form hydrogels. These hydrogels were characterized for rheological properties, tested with cultures of rat primary sensory neurons, and implanted subcutaneously in mice for 4 weeks. Sensory neurons cultured on high IKVAV concentration hydrogels (20%) formed longer neurite than those of neurons grown on hydrogels containing the scrambled IKVAV sequence. Finally, in vivo evaluation showed the absence of detectable inflammation. In conclusion, this functionalized ELP-IKVAV biomaterial shows interesting properties for tissue engineering requiring neurotization., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Efflux pumps are membrane protein complexes conserved in all living organisms. Beyond being involved in antibiotic extrusion in several bacteria, efflux pumps are emerging as relevant players in pathogen-host interactions. We have investigated on the possible role of the efflux pump network in Shigella flexneri, the etiological agent of bacillary dysentery. We have found that S. flexneri has retained 14 of the 20 pumps characterized in Escherichia coli and that their expression is differentially modulated during the intracellular life of Shigella. In particular, the emrKY operon, encoding an efflux pump of the Major Facilitator Superfamily, is specifically and highly induced in Shigella-infected U937 macrophage-like cells and is activated in response to a combination of high K + and acidic pH, which are sensed by the EvgS/EvgA two-component system. Notably, we show that following S. flexneri infection, macrophage cytosol undergoes a mild reduction of intracellular pH, permitting EvgA to trigger the emrKY activation. Finally, we present data suggesting that EmrKY is required for the survival of Shigella in the harsh macrophage environment, highlighting for the first time the key role of an efflux pump during the Shigella invasive process.

For patients with severe kidney or liver failure the best solution is currently organ transplantation. However, not all patients are eligible for transplantation and due to limited organ availability, most patients are currently treated with therapies using artificial kidney and artificial liver devices. These therapies, despite their relative success in preserving the patients' life, have important limitations since they can only replace part of the natural kidney or liver functions. As blood detoxification (and other functions) in these highly perfused organs is achieved by specialized cells, it seems relevant to review the approaches leading to bioengineered organs fulfilling most of the native organ functions. There, the culture of cells of specific phenotypes on adapted scaffolds that can be perfused takes place. In this review paper, first the functions of kidney and liver organs are briefly described. Then artificial kidney/liver devices, bioartificial kidney devices, and bioartificial liver devices are focused on, as well as biohybrid constructs obtained by decellularization and recellularization of animal organs. For all organs, a thorough overview of the literature is given and the perspectives for their application in the clinic are discussed., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Among the intestinal pathogenic Escherichia coli , enteroinvasive E. coli (EIEC) are a group of intracellular pathogens able to enter epithelial cells of colon, multiplicate within them, and move between adjacent cells with a mechanism similar to Shigella , the ethiological agent of bacillary dysentery. Despite EIEC belong to the same pathotype of Shigella , they neither have the full set of traits that define Shigella nor have undergone the extensive gene decay observed in Shigella . Molecular analysis confirms that EIEC are widely distributed among E. coli phylogenetic groups and correspond to bioserotypes found in many E. coli serogroups. Like Shigella , also in EIEC the critical event toward a pathogenic life-style consisted in the acquisition by horizontal gene transfer of a large F-type plasmid (pINV) containing the genes required for invasion, intracellular survival, and spreading through the intestinal mucosa. In Shigella , the ample gain in virulence determinants has been counteracted by a substantial loss of functions that, although important for the survival in the environment, are redundant or deleterious for the life inside the host. The pathoadaptation process that has led Shigella to modify its metabolic profile and increase its pathogenic potential is still in infancy in EIEC, although maintenance of some features typical of E. coli might favor their emerging relevance as intestinal pathogens worldwide, as documented by recent outbreaks in industrialized countries. In this review, we will discuss the evolution of EIEC toward Shigella -like invasive forms going through the epidemiology, including the emergence of new virulent strains, their genome organization, and the complex interactions they establish with the host.

In Pseudomonas aeruginosa , the ferric uptake regulator (Fur) protein controls both metabolism and virulence in response to iron availability. Differently from other bacteria, attempts to obtain fur deletion mutants of P. aeruginosa failed, leading to the assumption that Fur is an essential protein in this bacterium. By investigating a P. aeruginosa conditional fur mutant, we demonstrate that Fur is not essential for P. aeruginosa growth in liquid media, biofilm formation, and pathogenicity in an insect model of infection. Conversely, Fur is essential for growth on solid media since Fur-depleted cells are severely impaired in colony formation. Transposon-mediated random mutagenesis experiments identified pyochelin siderophore biosynthesis as a major cause of the colony growth defect of the conditional fur mutant, and deletion mutagenesis confirmed this evidence. Impaired colony growth of pyochelin-proficient Fur-depleted cells does not depend on oxidative stress, since Fur-depleted cells do not accumulate higher levels of reactive oxygen species (ROS) and are not rescued by antioxidant agents or overexpression of ROS-detoxifying enzymes. Ectopic expression of pch genes revealed that pyochelin production has no inhibitory effects on a fur deletion mutant of Pseudomonas syringae pv. tabaci , suggesting that the toxicity of the pch locus in Fur-depleted cells involves a P. aeruginosa -specific pathway(s). IMPORTANCE Members of the ferric uptake regulator (Fur) protein family are bacterial transcriptional repressors that control iron uptake and storage in response to iron availability, thereby playing a crucial role in the maintenance of iron homeostasis. While fur null mutants of many bacteria have been obtained, Fur appears to be essential in Pseudomonas aeruginosa for still unknown reasons. We obtained Fur-depleted P. aeruginosa cells by conditional mutagenesis and showed that Fur is dispensable for planktonic growth, while it is required for colony formation. This is because Fur protects P. aeruginosa colonies from toxicity exerted by the pyochelin siderophore. This work provides a functional basis to the essentiality of Fur in P. aeruginosa and highlights unique properties of the Fur regulon in this species., (Copyright © 2017 American Society for Microbiology.)

Introduction: Our objective is to describe the dietary habits, food preferences and adherence to Mediterranean diet (MeDi) of a large sample of Italian Parkinson's Disease (PD) patients compared to a group of controls., Methods: Dietary habits of 600 PD patients from throughout Italy and 600 controls matched by gender, age, education, physical activity level and geographical residence, were collected using the ON-GP Food Frequency Questionnaire. Then, we compared patients by disease duration and the presence of swallowing disturbances., Results: Overall, adherence of PD patients (males, 53.8%; mean disease duration, 9.2 ± 7.0 years) to MeDi was similar to controls (score, 4.8 ± 1.7 vs. 4.9 ± 1.6; P = 0.294). Patients consumed less alcohol and fish and drank significantly less water, coffee, and milk which resulted also in lower total fluids intake. On the contrary, they ate more fruit, cooked vegetables, cereals and baked items, more dressings and more sweets in general. Disease duration was associated with increased intake of several food groups but it was not associated with changes in MeDi score (P = 0.721). Patients with swallowing disturbances (n = 72) preferred softer and more viscous food but preferences did not result in differences in dietary pattern. However, patients with dysphagia drank less fluids (P = 0.043)., Discussion: PD patients presented different dietary habits and food preferences compared to the general population and adherence to MeDi was not associated with disease duration. Self-reported dysphagia was associated with reduced intake of fluids. These aspects may be amenable to change in order to improve the management of nutritional issues in this patient population., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background & Aims: Parkinson's disease (PD) patients can benefit considerably from appropriate nutritional care, particularly from diet. However, there is limited evidence on the eating habits of PD patients and their relationship with the features of the disease., Methods: We conducted a large case-control study. Consecutive PD patients (N = 600) receiving systematic nutritional care and healthy controls (N = 600) matched (1:1) for age, gender, education, physical activity level and residence were studied using a 66-item food frequency questionnaire. The relationship between dietary habits and the following features of PD were investigated in patients: body weight, energy balance, constipation, and levodopa therapy (dose) and its related motor complications., Results: PD patients had lower BMI and reported higher food intake than controls. BMI was found to be inversely associated with disease duration and severity, and levodopa-related motor complications, whereas energy intake was positively associated with these variables. An increase in protein intake by 10 g over physiological requirements (0.8 g/kg/day) corresponded to a mean increase in levodopa dose of 0.7 mg/kg/day. Constipation was also associated with higher levodopa requirements. Finally, protein intake and its distribution throughout the day influenced levodopa-related motor complications., Conclusion: The management of protein intake and the treatment of constipation should be considered to be an integral part of the care of PD patients. Attention should always be focused on energy intake also. This would result in the maintenance of nutritional status, the optimization of levodopa-therapy and the minimization of its related motor complications., (Copyright © 2016 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.)

Adhesive coatings for implantable biomaterials can be designed to prevent material displacement from the site of implant. In this paper, a strategy based on the use of melanin-like nanoparticles (MNPs) for the development of adhesive coatings for polysaccharidic membranes was devised. MNPs were synthesized in vitro and characterized in terms of dimensions and surface potential, as a function of pH and ionic strength. The in vitro biocompatibility of MNPs was investigated on fibroblast cells, while the antimicrobial properties of MNPs in suspension were evaluated on E. coli and S. aureus cultures. The manufacturing of the adhesive coatings was carried out by spreading MNPs over the surface of polysaccharidic membranes; the adhesive properties of the nano-engineered coating to the target tissue (intestinal serosa) were studied in simulated physiological conditions. Overall, this study opens for novel approaches in the design of naturally inspired nanostructured adhesive systems., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Shigella, the etiological agent of bacillary dysentery (shigellosis), is a highly adapted human pathogen. It evolved from an innocuous ancestor resembling the Escherichia coli strain by gain and loss of genes and functions. While the gain process concerns the acquisition of the genetic determinants of virulence, the loss is related to the adaptation of the genome to the new pathogenic status and occurs by pathoadaptive mutation of antivirulence genes. In this study, we highlight that the SRRz/Rz 1 lambdoid lysis cassette, even though stably adopted in E. coli K12 by virtue of its beneficial effect on cell physiology, has undergone a significant decay in Shigella. Moreover, we show the antivirulence nature of the SRRz/Rz 1 lysis cassette in Shigella. In fact, by restoring the SRRz/Rz 1 expression in this pathogen, we observe an increased release of peptidoglycan fragments, causing an unbalance in the fine control exerted by Shigella on host innate immunity and a mitigation of its virulence. This strongly affects the virulence of Shigella and allows to consider the loss of SRRz/Rz 1 lysis cassette as another pathoadaptive event in the life of Shigella., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Objectives: To estimate prevalence of sarcopenia and dynapenia in outpatients with Parkinson disease (PD) and to investigate their association with the features of the disease., Design: Cross-sectional study., Setting: A specialized tertiary care center., Participants: Consecutive patients (n = 364) aged 65 years or older, affected by parkinsonian syndromes., Measurements: Skeletal muscle mass (SMM), as well as strength and gait speed (GS) were assessed by bioimpedence analysis, handgrip dynamometry, and the 4-meter walking test, respectively. Based on these assessments, sarcopenia was diagnosed using the European Working Group on Sarcopenia in Older People criteria. Dynapenia was defined as handgrip strength less than 30 kg in men and less than 20 kg in women., Results: In total, 235 patients (64.6%) had a diagnosis of idiopathic PD. Low SMM index was recorded in 27 patients. Due to gait disturbances and postural instability, GS could not be measured in 98 patients and was found to be reduced in 61.3% of those assessed. Prevalence of sarcopenia and dynapenia was 6.6% (95% confidence interval [CI] 4.3-9.7) and 75.5% (95% CI 70.8-79.9), respectively. Sarcopenia tended to be higher in patients unable to perform GS assessment and was unrelated to the type of parkinsonian syndrome. It was associated with older age, longer disease duration, more severe disease, and higher disability in activities of daily living, as assessed by disease-specific clinical rating scale. Dynapenia was directly associated with parkinsonism other than PD, older age, and disability, whereas regular physical therapy appeared to be a preventive factor. However, it was unrelated to disease duration and severity. Finally, the disability score of activities of daily living was inversely correlated with handgrip strength and GS, whereas no association was found with SMM index., Conclusion: Being primarily motor disorders, parkinsonian syndromes are characterized by progressive disability in performing activities of daily living. Impaired functional status is a prominent feature of this patient population, independently of disease duration and severity. Sarcopenia is mainly related to advancing disease and, due to a significant sparing of SMM, is an infrequent condition, likely to play a minor role in disability. Several factors could be responsible for this favorable body composition (eg, motor symptoms, levodopa therapy) and deserve further investigation. The prognostic impact of sarcopenia also needs to be addressed., (Copyright © 2016 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.)

Background: Vasomotor rhinitis (VMR) and irritable bowel syndrome (IBS) are two of the most widespread pathologies in industrialized countries, and they have a substantial impact on health-related quality of life., Objective: To investigate the relationship between VMR and IBS to provide evidence to classify VMR and IBS as a sole disorder., Methods: The study included 150 patients from San Salvatore Hospital, L'Aquila: 50 with VMR, 50 with IBS, and 50 healthy volunteers., Results: With regard to a probable link between VMR and IBS, 62 patients of the whole population, 150 patients (41.3%) were affected by both VMR and IBS, 33 subjects (22%) were not affected by VMR or IBS, 25 patients (16.67%) only had VMR, and 30 patients (20%) only had IBS. When considering the 87 subjects affected by VMR, 62 of them also had IBS (71.26%). In turn, when considering all 92 subjects with IBS, 62 (67.39%) had VMR. We found a significant association among IBS and nasal obstruction, rhinorrhea, and turbinates, and among VMR and a change in stool consistency and abdominal pain. Among the patients affected by nonallergic rhinitis (NAR), we found a prevalence of NAR with eosinophils (31%) compared with the other cytologic types., Conclusion: This study expanded the knowledge of the link between VMR and IBS, also the correlation between typical VMR symptoms and IBS, and about the correlation between typical IBS symptoms and VMR. For future implications, the diagnosis of NAR indicated the need to check for the presence of IBS, by using the Rome III criteria, and a diagnosis of IBS indicated the need to check for the presence of VMR. More studies are needed to find the pathogenetic mechanisms to explain the clinical correlation between VMR and IBS as seen in the present study.

In Pseudomonas aeruginosa the Gac signaling system and the second messenger cyclic diguanylate (c-di-GMP) participate in the control of the switch between planktonic and biofilm lifestyles, by regulating the production of the two exopolysaccharides Pel and Psl. The Gac and c-di-GMP regulatory networks also coordinately promote the production of the pyoverdine siderophore, and the extracellular polysaccharides Pel and Psl have recently been found to mediate c-di-GMP-dependent regulation of pyoverdine genes. Here we demonstrate that Pel and Psl are also essential for Gac-mediated activation of pyoverdine production. A pel psl double mutant produces very low levels of pyoverdine and shows a marked reduction in the expression of the pyoverdine-dependent virulence factors exotoxin A and PrpL protease. While the exopolysaccharide-proficient parent strain forms multicellular planktonic aggregates in liquid cultures, the Pel and Psl-deficient mutant mainly grows as dispersed cells. Notably, artificially induced cell aggregation is able to restore pyoverdine-dependent gene expression in the pel psl mutant, in a way that appears to be independent of iron diffusion or siderophore signaling, as well as of recently described contact-dependent mechanosensitive systems. This study demonstrates that cell aggregation represents an important cue triggering the expression of pyoverdine-related genes in P. aeruginosa, suggesting a novel link between virulence gene expression, cell-cell interaction and the multicellular community lifestyle.

Muscle insulin resistance is linked to oxidative stress and decreased mitochondrial function. However, the exact cause of muscle insulin resistance is still unknown. Since offspring of patients with type 2 diabetes mellitus (T2DM) are susceptible to developing insulin resistance, they are ideal for studying the early development of insulin resistance. By using primary muscle cells derived from obese non-diabetic subjects with (FH+) or without (FH-) a family history of T2DM, we aimed to better understand the link between mitochondrial function, oxidative stress, and muscle insulin resistance. Insulin-stimulated glucose uptake and glycogen synthesis were normal in FH+ myotubes. Resting oxygen consumption rate was not different between groups. However, proton leak was higher in FH+ myotubes. This was associated with lower ATP content and decreased mitochondrial membrane potential in FH+ myotubes. Surprisingly, mtDNA content was higher in FH+ myotubes. Oxidative stress level was not different between FH+ and FH- groups. Reactive oxygen species content was lower in FH+ myotubes when differentiated in high glucose/insulin (25mM/150pM), which could be due to higher oxidative stress defenses (SOD2 expression and uncoupled respiration). The increased antioxidant defenses and mtDNA content in FH+ myotubes suggest the existence of compensatory mechanisms, which may provisionally prevent the development of insulin resistance., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We report a case of massive cerebellar infarction in which CT and MRI led to the diagnosis of acute hydrocephalus. All the neurological symptoms cleared after CSF shunting.

The authors report the preliminary results of a prospective study on intra-arterial chemotherapy and radiotherapy of advanced cervix carcinoma. The drugs were delivered by a totally implanted Infusaid Model 400 pump and patients received cyclic therapy consisting of Cis-Platinum, Methotrexate and Bleomycin. The catheter tips were positioned during a laparotomy in the hypogastric arteries with sutures and the pump was implanted in a subcutaneous pouch in the abdomen. After chemotherapy all patients had radiotherapy. Five patients entered in the study. Preliminary results suggest that long-term intra-arterial infusion chemotherapy for the treatment of advanced cervix carcinoma is practical for many patients. Two of five patients, after intra-arterial chemotherapy and radiotherapy, had radical hysterectomy.

256 patients (187 men and 69 women, aged 60 on the average) with carotid artery bruits, underwent digital subtraction angiography of the epiaortic vessels. The carotid artery stenoses of more than 50% and the carotid artery occlusions have been considered angiographically and compared with the evidences of the auscultation over the carotid bifurcation. The sensibility, specificity and overall accuracy of the carotid bruits were estimated. The sensibility was of 95%, the specificity of 83% and the overall accuracy of 92% when a carotid artery stenosis of more than 50% was detected. When a carotid occlusion was found the sensibility was of 71%, the specificity of 84% and the overall accuracy of 80%. When both the carotid stenosis of more than 50% and the carotid occlusion have been considered together, the sensibility was of 92%, the specificity of 84% and the overall accuracy of 90%. The Authors suggest that the carotid bruits are trustworthy witnesses of occlusive carotid artery disease.

Different dosages, varying from 250 to 500 mg/die, of ticlopidine per os have been employed in 46 patients (32 men and 14 women), average age 66, who had been admitted for acute non-embolic cerebral infarction. In the two clinical examinations, effected on average 5.4 and 18.3 days after the admission, significant alterations in 9 among the considered items have been noticed, i.e.: central deficit of the VIIth cranial nerve, motor function and gait impairment, Babinski's sign, loss of sensibility, dysarthria, visual field defect, cerebellar signs, motor and/or receptive dysphasia. A global improvement of these clinical signs has been noticed on average in 48.2% of the patients, while an exacerbation has occurred in 2.1% of the cases. None of the patients who have been considered in this study has decreased or has shown relapses of ischemic cerebral attacks during the observation. The treatment with ticlopidine has been discontinued in 3 cases (6.5%) for the occurrence of temporary adverse side-effects.

The results of the carotid artery Doppler ultrasound examination were compared with those of digital subtraction angiography in 256 patients (187 men and 69 women) with carotid bruit. The hemodynamically significant stenoses of more than 50% and the occlusions in the 512 carotid arteries examined were considered and the sensibility, the specificity and the overall accuracy of the Doppler ultrasound examination were calculated. When a carotid stenosis was diagnosed a sensibility of 97%, a specificity of 95% and an overall accuracy of 96% was found, while, for the diagnosis of carotid occlusion, the extimated sensibility, specificity and overall accuracy was 95%. Therefore, the ability to recognize a normal or an occluded carotid artery was of 95% (we found 125 out of 131 normal carotid arteries and 52 out of 55 occluded carotid arteries), whereas a stenotic carotid artery was identified in the 97% of cases (316 out of 326 stenotic carotid arteries). When both diagnoses of carotid stenosis and carotid occlusion were considered together, the extimated sensibility, specificity and overall accuracy were of 98%, 95% and 98% respectively. So the probability of misdiagnosing was of 2% (we made 10 wrong diagnoses out of 512 carotid arteries examined).