Role of HSV-1 in Alzheimer's disease pathogenesis: A challenge for novel preventive/therapeutic strategies

Herpes simplex virus-1 (HSV-1) is a ubiquitous DNA virus able to establish a life-long latent infection in host sensory ganglia. Following periodic reactivations, the neovirions usually target the site of primary infection causing recurrent diseases in susceptible individuals. However, reactivated HSV-1 may also reach the brain resulting in severe herpetic encephalitis or in asymptomatic infections. These have been reportedly linked to neurodegenerative disorders, such as Alzheimer's disease (AD), suggesting antiviral preventive or/therapeutic treatments as possible strategies to counteract AD onset and progression. Here, we provide an overview of the AD-like mechanisms driven by HSV-1-infection in neurons and discuss the ongoing trials repurposing anti-herpetic drugs to treat AD as well as preventive strategies aimed at blocking virus infection

Recurrent herpes simplex virus-1 infection induces hallmarks of neurodegeneration and cognitive deficits in mice

Herpes simplex virus type 1 (HSV-1) is a DNA neurotropic virus, usually establishing latent infections in the trigeminal ganglia followed by periodic reactivations. Although numerous findings suggested potential links between HSV-1 and Alzheimer's disease (AD), a causal relation has not been demonstrated yet. Hence, we set up a model of recurrent HSV-1 infection in mice undergoing repeated cycles of viral reactivation. By virological and molecular analyses we found: i) HSV-1 spreading and replication in different brain regions after thermal stress-induced virus reactivations; ii) accumulation of AD hallmarks including amyloid-\u3b2 protein, tau hyperphosphorylation, and neuroinflammation markers (astrogliosis, IL-1\u3b2 and IL-6). Remarkably, the progressive accumulation of AD molecular biomarkers in neocortex and hippocampus of HSV-1 infected mice, triggered by repeated virus reactivations, correlated with increasing cognitive deficits becoming irreversible after seven cycles of reactivation. Collectively, our findings provide evidence that mild and recurrent HSV-1 infections in the central nervous system produce an AD-like phenotype and suggest that they are a risk factor for AD

M2 receptors activation affects proliferation, migration and neurotrophic factors production in rat Schwann-like cells.

Introduction: Schwann cells (SCs) are key regulators of peripheral nerve regeneration and axonal myelination. Although SCs are an attractive therapeutic option for peripheral nerve injuries, there are several restrictions on their clinical application. Adipose derived stem cells (ASCs) represent an attractive source for cell therapies. When exposed to selective growth factors, they can acquire a SC-like phenotype (dASCs) expressing key SCs markers and adopting a spindle-shape morphology. Our group has demonstrated that Acetylcholine, via M2 muscarinic receptors, causes a reversible arrest of cell proliferation in SCs, increasing myelin proteins expression. dASCs also express muscarinic receptors. In the present work we evaluate if M2 muscarinic receptor activation may contribute to dASCs proliferation and phenotype. Methods: Cell growth was analysed by MTS assay. By ELISA and western blot analysis, two different NGF isoforms were detected and their release measured. Cell migration was analysed by wound healing assay. Results: M2 receptor activation causes a reversible arrest of the cell growth, without affecting cell survival. Moreover, they inhibit dASCs migration. M2 receptor stimulation also causes a significant decrease in neurotrophins expression, accompanied by a decreased release of proNGF and mNGF forms. In particular, the apoptotic proNGF isoform (25kDa) is strongly reduced after M2 receptor stimulation. Conclusions: Our data clearly demonstrates that M2 receptor activation inhibits dASCs proliferation and migration, negatively modulating neurotrophins expression and maturation. This indicates that M2 receptors may be relevant for dASCs maturation and myelination, perhaps in a similar way to that observed in SCs

Electroacupuncture in rats normalizes the diabetes-induced alterations in the septo-hippocampal cholinergic system

Diabetes induces early sufferance in the cholinergic septo-hippocampal system, characterized by deficits in learning andmemory, reduced hippocampal plasticity and abnormal pro-nerve growth factor (proNGF) release from hippocampal cells, all linked to dysfunctions in the muscarinic cholinergic modulation of hippocampal physiology. These alterations are associated with dysregulation of several cholinergic markers, such as the NGF receptor system and the acetylcholine biosynthetic enzyme choline-acetyl transferase (ChAT), in the medial septum and its target, the hippocampus. Controlled and repeated sensory stimulation by electroacupuncture has been proven effective in counteracting the consequences of diabetes on cholinergic system physiology in the brain. Here, we used a wellestablished Type 1 diabetes model, obtained by injecting young adult male rats with streptozotocin, to induce sufferance in the septo-hippocampal system.We then evaluated the effects of a 3-week treatmentwith low-frequency electroacupuncture on: (a) the expression and protein distribution of proNGF in the hippocampus, (b) the tissue distribution and content of NGF receptors in the medial septum, (c) the neuronal cholinergic and glial phenotype in the septo-hippocampal circuitry. Twice-a-week treatment with low-frequency electroacupuncture normalized, in both hippocampus and medial septum, the ratio between the neurotrophic NGF and its neurotoxic counterpart, the precursor proNGF. Electroacupuncture regulated the balance between the two major proNGF variants (proNGF-A and proNGF-B) at both gene expression and protein synthesis levels. In addition, electroacupuncture recovered to basal level the pro-neurotrophic NGF receptor tropomyosin receptor kinase-A content, down-regulated in medial septum cholinergic neurons by diabetes. Electroacupuncture also regulated ChAT content in medial septum neurons and its anterograde transport toward the hippocampus. Our data indicate that repeated sensory stimulation can positively affect brain circuits involved in learning and memory, reverting early impairment induced by diabetes development. Electroacupuncture could exert its effects on the septo-hippocampal cholinergic neurotransmission in diabetic rats, not only by rescuing the hippocampal muscarinic responsivity, as previously described, but also normalizing acetylcholine biosynthesis and NGFmetabolismin the hippocampus

Innovative mouse model mimicking human-like features of spinal cord injury: efficacy of Docosahexaenoic acid on acute and chronic phases

Traumatic spinal cord injury has dramatic consequences and a huge social impact. We propose a new mouse model of spinal trauma that induces a complete paralysis of hindlimbs, still observable 30 days after injury. The contusion, performed without laminectomy and deriving from the pressure exerted directly on the bone, mimics more closely many features of spinal injury in humans. Spinal cord was injured at thoracic level 10 (T10) in adult anesthetized female CD1 mice, mounted on stereotaxic apparatus and connected to a precision impactor device. Following severe injury, we evaluated motor and sensory functions, and histological/morphological features of spinal tissue at different time points. Moreover, we studied the effects of early and subchronic administration of Docosahexaenoic acid, investigating functional responses, structural changes proximal and distal to the lesion in primary and secondary injury phases, proteome modulation in injured spinal cord. Docosahexaenoic acid was able i) to restore behavioural responses and ii) to induce pro-regenerative effects and neuroprotective action against demyelination, apoptosis and neuroinflammation. Considering the urgent health challenge represented by spinal injury, this new and reliable mouse model together with the positive effects of docosahexaenoic acid provide important translational implications for promising therapeutic approaches for spinal cord injuries

Different responses of PC12 cells to different pro-nerve growth factor protein variants

The present work aimed to explore the innovative hypothesis that different transcript/protein variants of a pro-neurotrophin may generate different biological outcomes in a cellular system. Nerve growth factor (NGF) is important in the development and progression of neurodegenerative and cancer conditions. Mature NGF (mNGF) originates from a precursor, proNGF, produced in mouse in two major variants, proNGF-A and proNGF-B. Different receptors bind mNGF and proNGF, generating neurotrophic or neurotoxic outcomes. It is known that dysregulation in the proNGF/mNGF ratio and in NGF-receptors expression affects brain homeostasis. To date, however, the specific roles of the two major proNGF variants remain unexplored. Here we attempted a first characterization of the possible differential effects of proNGF-A and proNGF-B on viability, differentiation and endogenous ngf gene expression in the PC12 cell line. We also investigated the differential involvement of NGF receptors in the actions of proNGF. We found that native mouse mNGF, proNGF-A and proNGF-B elicited different effects on PC12 cell survival and differentiation. Only mNGF and proNGF-A promoted neurotrophic responses when all NGF receptors are exposed at the cell surface. Tropomyosine receptor kinase A (TrkA) blockade inhibited cell differentiation, regardless of which NGF was added to culture media. Only proNGF-A exerted a pro-survival effect when TrkA was inhibited. Conversely, proNGF-B exerted differentiative effects when the p75 neurotrophin receptor (p75NTR) was antagonized. Stimulation with NGF variants differentially regulated the autocrine production of distinct proNgf mRNA. Overall, our findings suggest that mNGF and proNGF-A may elicit similar neurotrophic effects, not necessarily linked to activation of the same NGF-receptor, while the action of proNGF-B may be determined by the NGF-receptors balance. Thus, the proposed involvement of proNGF/NGF on the development and progression of neurodegenerative and tumor conditions may depend on the NGF-receptors balance, on specific NGF trancript expression and on the proNGF protein variant ratio

Herpes Simplex Virus-1 in the Brain: The Dark Side of a Sneaky Infection

Herpes simplex virus-1 (HSV-1) establishes latency preferentially in sensory neurons of peripheral ganglia. A variety of stresses can induce recurrent reactivations of the virus, which spreads and then actively replicates to the site of primary infection (usually the lips or eyes). Viral particles produced following reactivation can also reach the brain, causing a rare but severe form of diffuse acute infection, namely herpes simplex encephalitis. Most of the time, this infection is clinically asymptomatic. However, it was recently correlated with the production and accumulation of neuropathological biomarkers of Alzheimer's disease. In this review we discuss the different cellular and molecular mechanisms underlying the acute and long-term damage caused by HSV-1 infection in the brain

Identification of Races 0 and 2 of Cladosporium fulvum (syn Passalora fulva) on Tomato in the Cinturón Hortícola de La Plata, Argentina

Surveys aimed at evaluating the incidence and severity of a new disease that developed in greenhouses cultivated with tomato (Solanum lycopersicum L.) were performed during 2009 and 2010 in greenhouses of the cultivars Elpida (Enza Zaden) and Colibrí (Clause) in an area of tomato production known as the Cinturón Hortícola de La Plata (the “horticultural belt of La Plata”). The disease had a 100% prevalence and 90% incidence within the ten 250 m2 greenhouses that were monitored in 2009, 2010, and 2011. In two consecutive assays, severity was 40%. The wide distribution of the disease suggests that the tomato hybrids under use lack resistance genes. The upper surface of diseased leaves had pale green to yellowish, 1- to 1.5-cm spots with undefined margins that progressed to a yellowish brown color, while on the lower side they had pale brown to brown sporulation of fungal conidiophores and conidia. Monosporic fungal cultures were obtained by needle transfer of conidia from sporulating areas of leaves (n = 20) to water agar medium. On 2% potato dextrose agar (PDA) the colonies of the relatively low growing fungus were strongly pigmented, greenish grey, and black on the reverse of the plate. The fungus developed one-celled, pale olive-green, ovoid conidia on long, branched chains, which arose from pigmented conidiophores, corresponding to the description of Cladosporium fulvum made by Joosten and de Wit (1). The identity of two isolates was confirmed by amplifying the 580-bp ITS sequences by means of primers ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) and ITS5 (5′-GAATTCGGAAGTAAAAGTCGTAACAAGG-3′) (ITS sequence Race 0 JQ768324.1 and Race 2 JQ768325.1). Both were 100% homologous to the ITS sequences of C. fulvum strains ATCC44962 (AF393700) and ATCC44960 (AF303701). Monosporic cultures of four isolates, each obtained from leaves collected from different plants growing in different greenhouses, were inoculated on a set of differential genotypes of tomato: cvs. Moneymaker, Cf-0, Cf-2, Cf-4, Cf-5, and Cf-9 (kindly provided by the Laboratory of Phytopathology of Wageningen University). Three plants of each tomato genotype at the 5 to 6 true leaf stage were inoculated by spraying a 105 conidia/ml conidial suspension of C. fulvum on the leaflets of the 3rd and 4th leaf. Inoculation tests of each isolate were repeated at least twice. After inoculation, plants were grown in the greenhouse at 13 to 29°C and 99% relative humidity. However, for the first 20 h after inoculation, plants were kept in the dark. They were regularly monitored and were scored as resistant or susceptible at 20 days after inoculation. Susceptible genotypes developed pale green to yellow spots on the abaxial leaf surface and pale brown to olivaceous brown sporulation on the adaxial side. Plants lacking disease symptoms were considered resistant. Inoculated fungi were reisolated from infected tissue and the identity of the fungal cultures confirmed based on morphology and the ITS sequence. Based on the reactions of the tomato genotypes, two races were identified, three isolates (race 2) developed symptoms only in cv. MM Cf-2, while the remaining isolate (race 0) provoked symptoms only in cv. MM Cf-0.Fil: Rollan, Cristina. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Protto, V. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Medina, Rocio. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: López, Silvina Marianela Yanil. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Vera Bahima, José. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Ronco, Blanca L.. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Saparrat, Mario Carlos Nazareno. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Balatti, Pedro Alberto. Universidad Nacional de la Plata. Facultad de Ciencias Agrarias y Forestales. Departamento de Ciencias Biológicas. Centro de Investigaciones de Fitopatología. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigaciones de Fitopatología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentin

Effects of intranasally-delivered pro-nerve growth factors on the septo-hippocampal system in healthy and diabetic rats

Pro-nerve growth factor (proNGF) is the predominant form of NGF in the brain and its levels increase in neurodegenerative diseases. The balance between NGF receptors may explain the contradictory biological activities of proNGF. However, the specific role of the two main proNGF variants is mostly unexplored. proNGF-A is prevalently expressed in healthy brain, while proNGF-B content increases in the neuro-degenerating brain. Recently we have investigated in vitro the biological action of native mouse proNGF variants. To gain further insights into the specific functions of the two proNGFs, here we intranasally delivered mouse-derived proNGF-A and proNGF-B to the brain parenchyma of healthy and diabetic rats, the latter characterized by dysfunction in spatial learning and memory, in the septo-hippocampal circuitry and by relative increase in proNGF-B hippocampal levels. Exogenous proNGF-B induces depression of hippocampal DG-LTP and impairment of hippocampal neurogenesis in healthy animals, with concomitant decrease in basal forebrain cholinergic neurons and cholinergic fibers projecting to the hippocampus. proNGF-A, while ineffective in healthy animals, rescues the diabetes-induced impairment in DG-LTP and hippocampal neurogenesis, promoting the concomitant recovery of the basal forebrain cholinergic phenotype. Our experimental evidences suggest that the balance between different proNGFs may influence the development and progression of neurodegenerative diseases