Papillomas and probable in situ carcinoma in association with a novel papillomavirus in a red-billed gull (Chroicocephalus novaehollandiae scopulinus).

Numerous raised plaques were observed on the feet of a red-billed gull (Chroicocephalus novaehollandiae scopulinus) that had been found dead. The plaques consisted of thickened epidermis with cell changes indicative of papillomavirus (PV) infection prominent within affected areas. Evidence suggesting progression to neoplasia was visible in one lesion. A DNA sequence that was most similar, but only 68.3% identical, to duck PV type 3 was amplified from the papillomas, suggesting a novel PV type. Lesions containing PV DNA have only previously been reported in three avian species. This is the first evidence that PVs could cause neoplasia in birds.

Adeno-associated Virus Vectors Efficiently Transduce Mouse and Rabbit Sensory Neurons Coinfected with Herpes Simplex Virus 1 following Peripheral Inoculation.

UNLABELLED: Following infection of epithelial tissues, herpes simplex virus 1 (HSV-1) virions travel via axonal transport to sensory ganglia and establish a lifelong latent infection within neurons. Recent studies have revealed that, following intraganglionic or intrathecal injection, recombinant adeno-associated virus (rAAV) vectors can also infect sensory neurons and are capable of stable, long-term transgene expression. We sought to determine if application of rAAV to peripheral nerve termini at the epithelial surface would allow rAAV to traffic to sensory ganglia in a manner similar to that seen with HSV. We hypothesized that footpad or ocular inoculation with rAAV8 would result in transduction of dorsal root ganglia (DRG) or trigeminal ganglia (TG), respectively. To test this, we inoculated the footpads of mice with various amounts of rAAV as well as rAAV capsid mutants. We demonstrated that this method of inoculation can achieve a transduction rate of >90% of the sensory neurons in the DRG that innervate the footpad. Similarly, we showed that corneal inoculation with rAAV vectors in the rabbit efficiently transduced >70% of the TG neurons in the optic tract. Finally, we demonstrated that coinfection of mouse footpads or rabbit eyes with rAAV vectors and HSV-1 resulted in colocalization in nearly all of the HSV-1-positive neurons. These results suggest that rAAV is a useful tool for the study of HSV-1 infection and may provide a means to deliver therapeutic cargos for the treatment of HSV infections or of dysfunctions of sensory ganglia. IMPORTANCE: Adeno-associated virus (AAV) has been shown to transduce dorsal root ganglion sensory neurons following direct intraganglionic sciatic nerve injection and intraperitoneal and intravenous injection as well as intrathecal injection. We sought to determine if rAAV vectors would be delivered to the same sensory neurons that herpes simplex virus (HSV-1) infects when applied peripherally at an epithelial surface that had been treated to expose the underlying sensory nerve termini. For this study, we chose two well-established HSV-1 infection models: mouse footpad infection and rabbit ocular infection. The results presented here provide the first description of AAV vectors transducing neurons following delivery at the skin/epithelium/eye. The ability of AAV to cotransduce HSV-1-infected neurons in both the mouse and the rabbit provides an opportunity to experimentally explore and disrupt host and viral proteins that are integral to the establishment of HSV-1 latency, to the maintenance of latency, and to reactivation from latency in vivo.

Relief of pain induced by varicella-zoster virus in a rat model of post-herpetic neuralgia using a herpes simplex virus vector expressing enkephalin.

Acute and chronic pain (post-herpetic neuralgia or PHN) are encountered in patients with herpes zoster that is caused by reactivation of varicella-zoster virus (VZV) from a state of neuronal latency. PHN is often refractory to current treatments, and additional strategies for pain relief are needed. Here we exploited a rat footpad model of PHN to show that herpes simplex virus (HSV) vector-mediated gene delivery of human preproenkephalin (vHPPE) effectively reduced chronic VZV-induced nocifensive indicators of pain. VZV inoculated at the footpad induced prolonged mechanical allodynia and thermal hyperalgesia that did not develop in controls or with ultraviolet light-inactivated VZV. Subsequent footpad administration of vHPPE relieved VZV-induced pain behaviors in a dose-dependent manner for extended periods, and prophylactic vector administration prevented VZV-induced pain from developing. Short-term pain relief following low-dose vHPPE administration could be effectively prolonged by vector re-administration. HPPE transcripts were increased three- to fivefold in ipsilateral ganglia, but not in the contralateral dorsal root ganglia. VZV hypersensitivity and its relief by vHPPE were not affected by peripheral delivery of opioid receptor agonist or antagonist, suggesting that the efficacy was mediated at the ganglion and/or spinal cord level. These results support further development of ganglionic expression of enkephalin as a novel treatment for the pain associated with Zoster.

Spread of herpes simplex virus to the spinal cord is independent of spread to dorsal root ganglia.

Levels of herpes simplex virus 1 (HSV-1) and HSV-2 DNA in dorsal root ganglia (DRG) and spinal cord (SC) were quantified after inoculation of guinea pig genitals and footpads. In genital infection, viral DNA reached SC and DRG simultaneously (at 2 to 3 days after inoculation) but was more abundant in SC than in DRG. After inoculation of footpads, which lack parasympathetic innervation, the viruses spread more efficiently to DRG than to SC. These results show important differences between genital and footpad infections, including independence of spread to DRG and SC, and imply that autonomic neurons may play an important role in the pathogenesis of viral latency after genital inoculation.

Chikungunya virus arthritis in adult wild-type mice.

Chikungunya virus is a mosquito-borne arthrogenic alphavirus that has recently reemerged to produce the largest epidemic ever documented for this virus. Here we describe a new adult wild-type mouse model of chikungunya virus arthritis, which recapitulates the self-limiting arthritis, tenosynovitis, and myositis seen in humans. Rheumatic disease was associated with a prolific infiltrate of monocytes, macrophages, and NK cells and the production of monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor alpha (TNF-alpha), and gamma interferon (IFN-gamma). Infection with a virus isolate from the recent Reunion Island epidemic induced significantly more mononuclear infiltrates, proinflammatory mediators, and foot swelling than did an Asian isolate from the 1960s. Primary mouse macrophages were shown to be productively infected with chikungunya virus; however, the depletion of macrophages ameliorated rheumatic disease and prolonged the viremia. Only 1 microg of an unadjuvanted, inactivated, whole-virus vaccine derived from the Asian isolate completely protected against viremia and arthritis induced by the Reunion Island isolate, illustrating that protection is not strain specific and that low levels of immunity are sufficient to mediate protection. IFN-alpha treatment was able to prevent arthritis only if given before infection, suggesting that IFN-alpha is not a viable therapy. Prior infection with Ross River virus, a related arthrogenic alphavirus, and anti-Ross River virus antibodies protected mice against chikungunya virus disease, suggesting that individuals previously exposed to Ross River virus should be protected from chikungunya virus disease. This new mouse model of chikungunya virus disease thus provides insights into pathogenesis and a simple and convenient system to test potential new interventions.

Novel betapapillomavirus associated with hand and foot papillomas in a cynomolgus macaque.

Betapapillomavirus is a genus of papillomaviruses (PVs) commonly found in human skin and associated with both benign and malignant skin lesions. Only 2 previous beta-PVs have been fully characterized in nonhuman species. This report describes a novel beta-PV, named Macaca fascicularis PV type 2 (MfPV2), isolated from exophytic skin papillomas on the hands and feet of a 2-year-old male cynomolgus monkey (M. fascicularis). On histology the papillomas were composed of diffusely thickened epidermis with superficial foci of cytomegaly, cytoplasmic pallor, marginalized chromatin, and rare eosinophilic intranuclear inclusion bodies. Positive immunostaining for p16 and the proliferation marker Ki67 was present multifocally within affected epidermis, most prominently within basal-type cells. Complete sequence identity (100%) was noted between PV genomes fully sequenced from hand and foot lesions. The MfPV2 genome was 7632 base pairs in length and included putative open reading frames (ORFs) for E1, E2, E4, E6, E7, L1, and L2 genes, similar to other PVs. The closest relatives to MfPV2 based on the L1 ORF sequence were all beta-PVs. These included human PV (HPV) 9, HPV115, HPV76, HPV75, and MfPV1 (60-70% pairwise identity for all), the latter of which was also isolated from hand and foot papillomas in a cynomolgus macaque. Phylogenetic analysis placed MfPV2 in a new species group (beta-6), distinct from HPVs (beta-1 to beta-5) and MfPV1 (beta-1). These findings characterize a new nonhuman beta-PV and provide additional support for the idea that tissue tropism among ancestral primate PVs developed prior to divergence of certain Old World primate lineages.

Contribution of rearranged actin structures to the spread of Ectromelia virus infection in vitro.

We describe here a contribution of virus-induced actin tails and filopodia in transmission of Ectromelia virus (ECTV) infection in permissive cells detected by the immunofluorescence and confocal microscopy. Immunoblot analysis revealed profoundly decreased beta-actin levels during ECTV replicative cycle in the infected cells 24 hrs post infection (p.i.). These results provided a basis for the further analysis of ECTV motion in the infected cells as well as for impact of ECTV infection on the cytoskeletal proteins.

Reduction in severity of a herpes simplex virus type 1 murine infection by treatment with a ribozyme targeting the UL20 gene RNA.

Hammerhead ribozymes were designed to target mRNA of several essential herpes simplex virus type 1 (HSV-1) genes. A ribozyme specific for the late gene U(L)20 was packaged in an adenovirus vector (Ad-U(L)20 Rz) and evaluated for its capacity to inhibit the viral replication of several HSV-1 strains, including that of the wild-type HSV-1 (17syn+ and KOS) and several acycloguanosine-resistant strains (PAAr5, tkLTRZ1, and ACGr4) in tissue culture. The Ad-U(L)20 Rz was also tested for its ability to block an HSV-1 infection, using the mouse footpad model. Mouse footpads were treated with either the Ad-U(L)20 Rz or an adenoviral vector expressing green fluorescent protein (Ad-GFP) and then infected immediately thereafter with 10(4) PFU of HSV-1 strain 17syn+. Ad-U(L)20 ribozyme treatment consistently led to a 90% rate of protection for mice from lethal HSV-1 infection, while the survival rate in the control groups was less than 45%. Consistent with this protective effect, treatment with the Ad-U(L)20 Rz reduced the viral DNA load in the feet, the dorsal root ganglia, and the spinal cord relative to that of the Ad-GFP-treated animals. This study suggests that ribozymes targeting essential genes of the late kinetic class may represent a new therapeutic strategy for inhibiting HSV infection.

Assessment of cryotherapy by liquid nitrogen in the treatment of hand and feet warts.

BACKGROUND: Cryotherapy with liquid nitrogen is the most widely used method to treat hand and feet warts. Therapeutic response to this method depends on many factors related to warts and to the patient. The aim of this study is to determine factors influencing therapeutic response of warts to cryotherapy by liquid nitrogen. METHODS: It was a prospective transversal study including 100 patients with warts of the hands and/or feet treated by cryotherapy with liquid nitrogen (cotton wool bud) and referred to Dermatology Department of Charles Nicolle Hospital of Tunis. Demographic data, so as characteristics of warts were recorded. Patients received one treatment/week with a maximum of 4 sessions. Patients whose warts were seen to be resolved were classified as cured. Cure-predictive factors were studied with a multi varied study with logistic regression. RESULTS: Of the 100 patients (56 females/44 males, Mean age: 22 years), ten were withdrawn. In 89 patients, warts were present on hands, whereas 23 had warts on feet and 12 had warts on both hands and feet. The mean number of warts per patient was 7. The total cure rate was of 64.4% and was more elevated in hands compared to feet (70.8% versus 10.5%). There was no difference between mean ages of cured group and not cured one (22.2 years versus 21 years). The mean duration of warts in cured patients was lower than that of not cured patients. The mean number of warts before treatment was 4.3 in cured patients and 12.3 warts in not cured patients. The mean number of treatments was 2.3 in cured patients and 4 treatments in not cured patients. The difference between these factors into cured and not cured groups was statistically significant in uni-varied study but not significant in multi-varied one. CONCLUSION: The effectiveness of liquid nitrogen used by traditional method in the treatment of hand and feet warts seems to depend on multiple factors: wart's duration, number of warts and number of treatments. These factors depend on each other.

Mittens and Booties Syndrome: A Unique Manifestation of Human Parechovirus Infection in Infants.

We describe the first 2 cases from the United States, of human parechovirus infection in infants manifesting a distinct rash of the hands and feet. We propose the term "Mittens and Booties Syndrome" and provide a review of the literature of all published cases.

Predicting the hand, foot, and mouth disease incidence using search engine query data and climate variables: an ecological study in Guangdong, China.

OBJECTIVES: Hand, foot, and mouth disease (HFMD) has caused a substantial burden in China, especially in Guangdong Province. Based on the enhanced surveillance system, we aimed to explore whether the addition of temperate and search engine query data improves the risk prediction of HFMD. DESIGN: Ecological study. SETTING AND PARTICIPANTS: Information on the confirmed cases of HFMD, climate parameters and search engine query logs was collected. A total of 1.36 million HFMD cases were identified from the surveillance system during 2011-2014. Analyses were conducted at aggregate level and no confidential information was involved. OUTCOME MEASURES: A seasonal autoregressive integrated moving average (ARIMA) model with external variables (ARIMAX) was used to predict the HFMD incidence from 2011 to 2014, taking into account temperature and search engine query data (Baidu Index, BDI). Statistics of goodness-of-fit and precision of prediction were used to compare models (1) based on surveillance data only, and with the addition of (2) temperature, (3) BDI, and (4) both temperature and BDI. RESULTS: A high correlation between HFMD incidence and BDI (r=0.794, p<0.001) or temperature (r=0.657, p<0.001) was observed using both time series plot and correlation matrix. A linear effect of BDI (without lag) and non-linear effect of temperature (1 week lag) on HFMD incidence were found in a distributed lag non-linear model. Compared with the model based on surveillance data only, the ARIMAX model including BDI reached the best goodness-of-fit with an Akaike information criterion (AIC) value of -345.332, whereas the model including both BDI and temperature had the most accurate prediction in terms of the mean absolute percentage error (MAPE) of 101.745%. CONCLUSIONS: An ARIMAX model incorporating search engine query data significantly improved the prediction of HFMD. Further studies are warranted to examine whether including search engine query data also improves the prediction of other infectious diseases in other settings.

Suramin treatment reduces chikungunya pathogenesis in mice.

The chikungunya virus (CHIKV), an arthritogenic alphavirus, has caused explosive epidemics involving millions of cases. Globally expanding pandemics involving CHIKV and post-CHIKV rheumatic disorders are increasing public health concerns. However, no antiviral interventions or vaccines to control CHIKV infection have yet been approved. Although suramin has been possess anti-CHIKV activity in vitro, whether suramin has anti-CHIKV activity in vivo remains unknown. This study aimed to determine whether suramin treatment could ameliorate CHIKV-induced arthritis in a C57BL/6 mice model. C57BL/6 mice were infected with CHIKVs to evaluate anti-CHIKV activities of suramin in terms of histopathology, viral burden and disease score. Not only did suramin treatment substantially decrease viral loads, but it also significantly ameliorated acute foot lesions in mice. In addition, suramin treatment markedly restores cartilage integrity and reduces the number of IHC positive chondrocyte in mice infected with CHIKV strains 0810bTw and 0706aTw. This in vivo study highlights the potential ability of suramin to treat CHIKV infection in clinical settings.

Avian reovirus replication in mononuclear phagocytes in chicken footpad and spleen after footpad inoculation.

Circulating monocytes and tissue macrophages were suggested to be susceptible to avian reovirus (ARV) infection. To determine if ARV infects and replicates in mononuclear phagocytes (KUL01-positive cells), we infected 3-day-old specific-pathogen-free chickens with ARV strain 2408 by inoculation of the left footpad. The left footpads and spleens were collected for analysis at 1.5 and 2.5 d after inoculation. Replication of ARV in the footpad and spleen was demonstrated by detection of the viral protein σNS using immunohistochemical testing and viral S1 RNA expression by real-time quantitative polymerase chain reaction (qPCR). Furthermore, immunofluorescent double-staining assay of cytocentrifuged cells and cryosections of the footpad and spleen for the viral protein σNS and the surface marker recognized by monoclonal antibody (MAb) KUL01 indicated that KUL01-positive cells costained with MAb H1E1, which recognizes ARV protein σNS. In addition, more ARV S1 RNA was measured by qPCR in the KUL01-positive cell samples prepared from the footpad or spleen 1.5 d after inoculation compared with non-KUL01-positive cell samples. The amounts of ARV S1 RNA in the spleen were significantly lower (P < 0.05) than the amounts in the footpad 1.5 d after inoculation. The results suggest that ARV infects mononuclear phagocytes and then replicates within these cells before migrating to the spleen, where it infects and replicates in KUL01-positive cells.

Il a été suggéré que les monocytes circulants et les macrophages tissulaires étaient sensibles à une infection par le reovirus aviaire (ARV). Afin de déterminer si l'ARV infecte et se réplique dans les phagocytes mononucléaires (cellules KUL01-positives), nous avons infecté des poussins exempts d'agents pathogènes spécifiques âgés de 3 j avec la souche 2408 d'ARV par inoculation dans le coussinet plantaire gauche. Les coussinets plantaires et les rates furent prélevés pour analyse aux jours 1,5 et 2,5 suivant l'inoculation. La réplication d'ARV dans le coussinet plantaire et la rate fut démontrée par détection de la protéine virale σNS par épreuve immunohistochimique et l'expression d'ARN S1 viral par réaction d'amplification en chaîne par la polymérase en temps réel (qPCR). De plus, l'épreuve d'immunofluorescence par double coloration de cellules cytocentrifugées et de coupes congelées du coussinet plantaire et de la rate pour la protéine virale σNS et le marqueur de surface reconnu par l'anticorps monoclonal (AcMo) KUL01 indiquait que les cellules positives pour KUL01se co-coloraient avec l'AcMo H1E1, qui reconnait la protéine σNS de l'ARV. Également, plus d'ARN S1 d'ARV était mesuré par qPCR dans les échantillons de cellules KUL01 positives préparés à partir de coussinets plantaires ou de rates 1,5 j après l'inoculation comparativement à des échantillons de cellules KUL01 négatives. Les quantités d'ARN S1 d'ARV dans la rate étaient significativement plus basses (P < 0,05) que les quantités dans les coussinets plantaires 1,5 j après l'inoculation. Les résultats suggèrent que l'ARV infecte les phagocytes mononucléaires et par la suite se répliquent dans ces cellules avant de migrer à la rate, où il infecte et se réplique dans les cellules KUL01-positives.(Traduit par Docteur Serge Messier).

Quantitation of foot-and-mouth disease virus genomes in bovine tissue by competitive RT-PCR.

The sensitivity of a reverse transcription-dependent polymerase chain reaction (RT-PCR) for detecting foot-and-mouth disease virus (FMDV) genomes was quantified by use of RNA transcribed in vitro from FMDV-specific cDNA. Previously, the cDNA had been elongated by 228 base pairs. The minimum number of template molecules required to obtain the specific RT-PCR product was determined to be 10(4). This was achieved by use of 1 microg of primer for cDNA synthesis and by undertaking of at least 30 cycles of PCR. Knowing the sensitivity of the system prompted the examination of clinical samples for content of FMDV genomes. The samples were tongue and foot epithelia as well as nasal discharge, removed 11-14 days after infection from 14 cattle. They all contained FMDV genomes but not in each clinical specimen. The size difference of the products amplified from transcript and viral genome enabled the estimate by competitive RT-PCR of the number of viral genomes contained in some samples. The RNA extracted from approximately 10(7) tissue cells each was found to contain between less than 10(6) and up to 10(8) FMDV genomes, irrespective of the sample type.

Immunohistochemical detection of canine distemper virus in haired skin, nasal mucosa, and footpad epithelium: a method for antemortem diagnosis of infection.

A reliable antemortem diagnostic method is needed for determining infection with canine distemper virus (CDV). The utility of immunohistochemical detection of CDV antigen was examined was examined for samples of nasal and footpad epithelium and haired skin in dogs with and without detectable CDV antigen in the lung and/or brain. Tissues from 57 dogs at risk of CDV infection were tested. Viral antigen was found in the lung and/or brain of 28 dogs. Among these dogs, viral antigen was demonstrated in the epithelial cells of the nasal mucosa in 24 of 27 dogs, in the footpad epithelium in 24 of 26 dogs, and in the haired skin of the dorsal neck in 26 of 27 dogs. Among the 29 dogs without CDV antigen in either the lung or brain, 1 dog had positive staining for viral antigen in the skin and nasal mucosa. Biopsies of haired skin of the dorsal neck, which is relatively simple to sample, can be used for immunohistochemical testing for acute and subacute infection with CDV.

Plantar wart treatment with combination imiquimod and salicylic acid pads.

Treatment of plantar warts is often difficult and may be painful, often employing destructive treatment modalities. We report the successful treatment of a patient with a large plantar wart using Imiquimod 5% cream under occlusion with a 40% salicylic acid pad. This combination treatment modality likely allows successful delivery of Imiquimod through the thick skin on the plantar surface. Once penetrated, an anti-viral state is created by upregulating specific cytokines to eradicate the human papilloma virus (HPV).

Plantar wart treatment with combination imiquimod and salicylic acid pads.

Treatment of plantar warts is often difficult and may be painful, often employing destructive treatment modalities. We report the successful treatment of a patient with a large plantar wart using Imiquimod 5% cream under occlusion with a 40% salicylic acid pad. This combination treatment modality likely allows successful delivery of Imiquimod through the thick skin on the plantar surface. Once penetrated, an anti-viral state is created by upregulating specific cytokines to eradicate the human papilloma virus (HPV).

Infectivity assays of foot-and-mouth disease virus: contact transmission between cattle and buffalo (Bubalus bubalis) in the early stages of infection.

No differences were observed between cattle and Indian buffalo (Bubalus bubalis) in terms of temperature, viraemia or virus replication in the pharyngeal area, during the acute phase of foot-and-mouth disease. Like cattle, the Indian buffalo became infected and excreted virus before any clinical signs of foot-and-mouth disease developed. The disease was transmitted from cattle to buffalo and vice versa, during the acute stage of infection, as if the animals had been of the same species, presumably because of their close phylogenetic relationship. There were more tongue lesions in the cattle than in the buffalo. Foot lesions in the buffalo at first had a scaley appearance, but later became vesicular. Anti-virus infection associated antigen and neutralising antibodies were synthesised at the same time in both species and reached similar titres in the same period. Persistent infection in the buffalo during the first 35 days after infection was similar to that in the cattle.

Spatio-temporal clustering of hand, foot, and mouth disease at the county level in Guangxi, China.

BACKGROUND: Amid numerous outbreaks of hand, foot and mouth disease (HFMD) in Asia over the past decade, studies on spatio-temporal clustering are limited. Without this information the distribution of severe cases assumed to be sporadic. We analyzed surveillance data with onset dates between 1 May 2008 to 31 October 2013 with the aim to document the spatio-temporal clustering of HFMD cases and severe cases at the county level. METHODS: Purely temporal and purely spatial descriptive analyses were done. These were followed by a space-time scan statistic for the whole study period and by year to detect the high risk clusters based on a discrete Poisson model. RESULTS: The annual incidence rate of HFMD in Guangxi increased whereas the severe cases peaked in 2010 and 2012. EV71 and CoxA16 were alternating viruses. Both HFMD cases and severe cases had a seasonal peak in April to July. The spatio-temporal cluster of HFMD cases were mainly detected in the northeastern, central and southwestern regions, among which three clusters were observed in Nanning, Liuzhou, Guilin city and their neighbouring areas lasting from 1.2 to 2.5 years. The clusters of severe cases were less consistent in location and included around 40-70% of all severe cases in each year. CONCLUSIONS: Both HFMD cases and severe cases occur in spatio-temporal clusters. The continuous epidemic in Nanning, Liuzhou, Guilin cities and their neighbouring areas and the clusters of severe cases indicate the need for further intensive surveillance.

[Viral warts on hands and feet are often self-limiting]. / Vorter på hænder og fødder er oftest selvlimiterende.

Viral warts are common skin lesions caused by human papilloma virus. This article describes the pathogenesis, symptoms and treatment methods of cutaneous warts. The majority resolves spontaneously and the evidence on treatment of warts is rather poor, however, products with salicylic acid and cryotherapy have been surveyed most rigorously and shown to increase treatment rates.