The olfactory bulb: A link between environmental agents and narcolepsy, from the standpoint of autoimmune etiology.

Narcolepsy type 1 is a lifelong sleep disorder characterized by the loss of hypocretin-producing neurons in the brain. Environmental agents, including influenza, neurotoxic metals, and combustion smoke, have been implicated in the pathogenesis, especially in carriers of the human leukocyte antigen class II DQB1*06:02 allele. Sensitive experimental approaches have recently revealed hypocretin-autoreactive CD4+ and CD8+ T cells in the blood of narcoleptic patients. However, such potentially harmful cells are also detectable, to a lesser degree, in control DQB1*06:02 carriers, suggesting that the integrity of the blood-brain barrier (BBB) provides a neuroprotective effect. Here, we present the hypothesis that external toxic agents induce neuroinflammation in the olfactory bulb and concomitant overproduction of proinflammatory cytokines (e.g., tumor necrosis factor-α and interferon-γ); this, in turn, compromises the BBB, allowing autoimmune cells to access and kill hypocretinergic neurons. Such sequential pathological alterations could occur insidiously, passing unnoticed and consequently being underestimated. The elevated number of autoreactive T cells in narcoleptics relative to controls might reflect externally induced immunomodulation rather than a direct disease trigger.

The olfactory bulb: A link between environmental agents and narcolepsy.

Narcolepsy with cataplexy is a lifelong sleep disorder associated with orexin/hypocretin deficiency in the central nervous system. In addition to a genetic predisposition, a variety of environmental factors, such as influenza viruses, have been implicated in the pathogenesis of the disease. In this article, a hypothesis is proposed that environmental agents access the olfactory bulb and trigger neuroinflammation, which in turn induces neurodegeneration of orexinergic neurons in the lateral hypothalamus and other neuronal subpopulations regulating the sleep-wake cycle, which triggers the development of narcolepsy.

Changes in Basic Movement Ability and Activities of Daily Living After Hip Fractures: Correlation Between Basic Movement Scale and Motor-Functional Independence Measure Scores.

OBJECTIVE: The aim of this study was to examine the correlation between basic movement ability and activities of daily living (ADL) in elderly patients after hip fracture surgery and predict ADL outcomes from changes in basic movement ability. DESIGN: Fifty-four patients receiving rehabilitation after hip fracture surgery were collected prospectively. Ambulatory ability was evaluated using a Basic Movement Scale (BMS), and ADL was evaluated using the motor subscale of the Functional Independence Measure (motor-FIM). From the results of evaluating BMS and motor-FIM weekly, the important postoperative period to regain ADL was investigated. RESULTS: There was a close correlation between BMS and motor-FIM scores at each evaluation point (r = 0.971, P < 0.001) and a significant correlation between weekly BMS and motor-FIM gains (r = 0.741, P < 0.001). Cluster analysis of BMS scores from postoperative week (POW) 2 to 12 showed three patterns of change, with BMS scores at POW 2 reflecting the outcome. CONCLUSIONS: The very strong correlation between BMS and motor-FIM scores suggests that BMS is a favorable indicator of changes in ADL. Because basic movement ability at POW 2 also reflected the prognosis, constructive interventions should be implemented early to help patients ambulate and regain other basic movements by no later than POW 2.

Olfactory vector hypothesis for encephalitis lethargica.

Viruses have long been implicated in the pathogenesis of classical encephalitis lethargica, which was first described by Constantin von Economo in 1917. In this article, I propose the hypothesis that an airborne virus travels along the olfactory conduit to infect the olfactory bulb; this local infection or induced neuroinflammation, in turn, retrogradely targets certain neuronal populations with sleep-wake regulatory functions in the hypothalamus and midbrain, leading to the development of wakeful inactivity, a hallmark clinical feature of the disease. Furthermore, the olfactory vector hypothesis may also explain the pathomechanism of the debilitating complication of the disease, i.e., postencephalitic parkinsonism, in terms of a recently discovered nigro-olfactory projection.

Viremic attack explains the dual-hit theory of Parkinson's disease.

The dual-hit theory of Parkinson's disease proposes that an airborne pathogen attacks both the olfactory and enteric nervous systems to initiate the Lewy pathology, gradually leading to devastating neurodegenerative processes within the brain. Based on published literatures, this article proposes a hypothesis that viruses with viremic potential can simultaneously attack both of these nervous systems via viremia due to the lack of a blood-nerve barrier in these tissues, thereby explaining the dual-hit theory. Understanding the precise mechanisms underlying the neuropathology will facilitate development of better prophylactic and early intervention strategies against Parkinson's disease.

Herpes simplex virus US3 protein kinase regulates host responses and determines neurovirulence.

The US3 of HSV encodes a serine/threonine protein kinase that is highly conserved among members of the alphaherpesviruses. It is an accessory gene that is not required for viral replication in cultured cells but appears essential for viral survival in humans. Although accumulating in vitro evidence suggested that the viral protein kinase is multifunctional, little information is available about its functions in vivo. Several reports point out that, upon invasion into the peripheral nervous system, HSV blocks virus-induced neuronal apoptosis, while presumably subverting host immune responses, largely through actions of the US3 protein kinase. In addition, the US3 protein kinase confers the viral neurovirulence. In the present article, functions of the HSV US3 protein kinase are briefly reviewed, with special attention given to its role in regulating host responses and neurovirulence.

Herpes simplex virus type 1 persists in the aged brain through hypothetical expression of accessory genes.

Herpes simplex virus type 1 persists in the brain of most aged individuals and may contribute to the pathogenesis of Alzheimer's disease. The virus likely utilizes accessory genes for neural spread within the nervous system and herpes simplex virus type 1 may regulate various host responses through an array of accessory genes. This mini-review focuses on these viral accessory genes that may shed light on the potential mechanisms of this enigmatic phenomenon in the elderly brain.

Interleukin-10 inhibits tumor necrosis factor-alpha production in lipopolysaccharide-stimulated RAW 264.7 cells through reduced MyD88 expression.

The mechanism of interleukin (IL)-10-mediated inhibition of tumor necrosis factor (TNF)-alpha production was studied by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. IL-10 inhibited TNF-alpha production transiently at an early stage after LPS stimulation. IL-10 inhibited the activation of nuclear factor (NF)-kappaB, p38 and stress-activated protein kinase (SAPK) in LPS-stimulated RAW 264.7 cells. Although the level of MyD88 protein increased in response to LPS, IL-10 prevented the LPS-induced MyD88 augmentation. There was no significant difference in the MyD88 mRNA expression between the cells pretreated with or without IL-10 in response to LPS. Therefore, IL-10 was suggested to inhibit LPS-induced TNF-alpha production via reduced MyD88 expression.

Evaluation of a virus derived from MDCK cells infected persistently with influenza A virus as a potential live-attenuated vaccine candidate in the mouse model.

A temperature-sensitive mutant virus unable to replicate at 38 degrees C was recovered from passage 189 (IVpi-189) of Madin-Darby canine kidney cells infected persistently with influenza A. Immunofluorescent staining of the IVpi-189 virus-infected cells revealed disrupted transport of the matrix (M) 1 protein into the nucleus at non-permissive temperatures, resulting in retention of the nucleoprotein (NP) in the nucleus. Upon comparison with the parental influenza A E61-24-P15 strain used to establish persistent infection, amino acid exchanges were found in the M1 protein of IVpi-189 virus; arginine to glutamine at position 72 and threonine to alanine at position 139. When mice were inoculated intranasally with IVpi-189 virus, virus growth in the lungs was restrained and terminated rapidly. Prior intranasal inoculation with only a small dose of IVpi-189 virus induced humoral and cellular immune responses and protected mice against subsequent virulent virus challenge. These results indicate that IVpi-189 virus, an avirulent temperature-sensitive mutant, is a promising candidate for use as a live-attenuated vaccine.

Oncolytic virotherapy for malignant melanoma with herpes simplex virus type 1 mutant HF10.

BACKGROUND: Many viruses have been engineered and evaluated for their potential as therapeutic agents in the treatment of malignant neoplasm, including malignant melanoma. OBJECTIVE: In this study, we investigated the efficacy of HF10, an attenuated, replication-competent HSV, in immunocompetent animal models with malignant melanoma. METHODS: For in vitro study, viral cytotoxicity assays and replication assays were performed both in human and mouse melanoma cells. For the study in vivo, intraperitoneally disseminated or subcutaneous melanoma models were prepared in DBA/2 mice using clone M3 cells, then HF10 was inoculated intraperitoneally or intratumorally. Therapeutic efficacy of HF10 was assessed by survival, tumor growth, and histopathological analysis. RESULTS: HF10 infection produced cytolytic effects in melanoma cells at various multiplicities of infection (MOI). In the intraperitoneal melanoma model, all mice survived when given intraperitoneal injections of HF10 compared with 100% fatality in the control mice. In the subcutaneous tumor model, intratumoral inoculation of HF10 significantly reduced tumor growth. Histology and immunohistochemistry showed tumor lysis and inflammatory cell infiltration after intratumoral HF10 inoculation. Viral antigen was retained at the inoculation site until 7 days post-infection. HF10-treated intraperitoneal tumor mice were also protected against tumor rechallenge. HF10 also affected the non-inoculated contralateral tumor when injected into the ipsilateral tumor of mice, suggesting that HF10 can induce systemic antitumor immune responses in mice. CONCLUSION: Oncolytic viral therapy using HF10 was effective in melanoma mouse models, and intratumoral injection of HF10 induced systemic antitumor responses. These results suggest that HF10 is a promising agent for the treatment of advanced melanoma.

Fetal calf serum inhibits virus genome expression in Madin-Darby canine kidney cells persistently infected with influenza A virus.

A cell line of Madin-Darby canine kidney (MDCK) cells persistently infected with human influenza A virus has been established and designated as MDCK-IVpi cells. Production of progeny virus in MDCK-IVpi cells was suppressed when the cells were incubated in the presence of 10% fetal calf serum (FCS). FCS impaired virus mRNA synthesis in MDCK-IVpi cells, which resulted in a scarcity of virus proteins for virion formation. However, MDCK-IVpi cells well supported the growth of superinfecting heterologous influenza viruses, even in the presence of FCS. A certain fetuin-like substance in FCS might be responsible for the observed inhibition of virus replication.

Influenza A virus derived from persistently virus-infected cells shows attenuated cytotoxicity in cultured cells but virulent pathogenicity in mice.

The IVpi-43 strain of influenza A virus, a progeny virus derived from persistently virus-infected Madin-Darby canine kidney (MDCK) cells, showed a more attenuated nature in cytopathology in cultured cells than the parental wild-type influenza virus (IVwt) that was used for establishment of the virus carrier culture. Upon infection of MDCK cells, growth of the IVpi-43 virus was restrained with an impaired synthesis of virus structural proteins in the cells. Apoptosis induced by IVpi-43 virus was confined at a low level. The IVpi-43 virus was able to easily cause persistent infection in fresh MDCK cells. In contrast to the in vitro phenotype, the IVpi-43 virus proved highly virulent in mice, with massive and broadly disseminated virus multiplication in the lungs. It was suggested that impaired activity of the neuraminidase molecule of the IVpi-43 virus was responsible for the delayed and faint appearance of apoptosis in the IVpi-43 virus-infected respiratory cells, which made it possible for the virus to replicate for a longer period and to spread to a broader area of the lungs and that abundant numbers of the virus-infected lung cells were killed within a short period by the subsequently established virus-specific immune responses, leading to unrecoverable serious pneumonia.

Differential expression of autophagy in Hodgkin lymphoma cells treated with various anti-cancer drugs.

AIM: autophagy is a pivotal physiological process for survival during starvation, differentiation and normal growth control. It is defined as the process of sequestrating cytoplasmic proteins or even entire organelles into the lytic compartment (lysosome/vacuole). This study investigates the expression of autophagy in Hodgkin lymphoma cells treated with various anti-cancer drugs. METHODS: Hodgkin's lymphoma cells (HD-My-Z cells) were cultured with various anti-cancer drugs, such as bleomycin, adriamycin, gemcitabine and paclitaxel. Autophagy was detected by fluorescent pattern of light chain 3(LC3) proteins and the apoptotic cell death was determined by annexin V binding. RESULTS: autophagy was detected in HD-My-Z cells treated with gemcitabine, but not with bleomycin, adriamycin and paclitaxel. Adriamycin exhibited the strongest cytotoxic action, and the cytotoxic action of bleomycin and gemcitabine was less marked compared with adriamycin. Paclitaxel did not cause significant cell death in the cells. CONCLUSION: autophagy was differentially expressed in Hodgkin lymphoma cells treated with anti-cancer drugs and the expression did not correspond to the apoptotic cell death.

Replication-competent, oncolytic herpes simplex virus type 1 mutants induce a bystander effect following ganciclovir treatment.

Cells expressing herpes simplex virus (HSV) thymidine kinase (tk) are killed by ganciclovir (GCV). Adjacent cells without HSV-tk also die, a phenomenon known as the 'bystander effect'. However, there is no evidence that replication-competent HSV induces a bystander effect in the presence of GCV. Therefore, we investigated the bystander effect in HEp-2 cells infected with replication-competent, oncolytic HSV-1 mutants, hrR3 and HF10. In cells infected at a multiplicity of infection (MOI) of 3, GCV did not induce apoptosis. At low MOIs of 0.3 and 0.03, however, a number of adjacent, uninfected cells apoptosed following GCV treatment. Irrespective of GCV treatment, HEp-2 cells expressed minimal levels of connexin 43 (Cx43). However, Cx43 expression was enhanced by GCV in response to infection with HF10 at an MOI of 0.3, but not at an MOI of 3. Expression of other proteins involved in gap junctions, including Cx26 and Cx40, was not augmented under these conditions. The PKA and PI3K signal transduction pathways are likely involved in enhanced Cx43 expression as inhibitors of these pathways prevented Cx43 upregulation. These results suggest that infection with replication-competent HSV-1 induces the bystander effect in cells treated with GCV because of efficient intercellular transport of active GCV through abundant gap junctions.

Herpes simplex virus protein UL11 but not UL51 is associated with lipid rafts.

The UL11 and UL51 gene products of herpes simplex virus (HSV) are membrane-associated tegument proteins that are incorporated into the HSV virion. UL11 and UL51 are conserved throughout the herpesvirus family. Both UL11 and UL51, either singly or in combination, are involved in virion envelopment and/or egress. Both proteins are fatty acylated: UL11 is both acylated by myristoic and palmitoic acids and UL51 is monoacylated by palmitoic acid. Using confocal microscopy and sucrose gradient fractionations in transfected or HSV-infected cells, we found that HSV-2 UL11 but not UL51 was associated with lipid rafts. The dual acylation of UL11 was necessary for lipid raft association, as mutations in the myristoylation or palmitoylation sites prevented lipid raft association. These differences in lipid raft association may contribute to the functional differences between UL11 and UL51.