Identification and epidemiological study of an uncultured flavivirus from ticks using viral metagenomics and pseudoinfectious viral particles.

During their blood-feeding process, ticks are known to transmit various viruses to vertebrates, including humans. Recent viral metagenomic analyses using next-generation sequencing (NGS) have revealed that blood-feeding arthropods like ticks harbor a large diversity of viruses. However, many of these viruses have not been isolated or cultured, and their basic characteristics remain unknown. This study aimed to present the identification of a difficult-to-culture virus in ticks using NGS and to understand its epidemic dynamics using molecular biology techniques. During routine tick-borne virus surveillance in Japan, an unknown flaviviral sequence was detected via virome analysis of host-questing ticks. Similar viral sequences have been detected in the sera of sika deer and wild boars in Japan, and this virus was tentatively named the Saruyama virus (SAYAV). Because SAYAV did not propagate in any cultured cells tested, single-round infectious virus particles (SRIP) were generated based on its structural protein gene sequence utilizing a yellow fever virus-based replicon system to understand its nationwide endemic status. Seroepidemiological studies using SRIP as antigens have demonstrated the presence of neutralizing antibodies against SAYAV in sika deer and wild boar captured at several locations in Japan, suggesting that SAYAV is endemic throughout Japan. Phylogenetic analyses have revealed that SAYAV forms a sister clade with the Orthoflavivirus genus, which includes important mosquito- and tick-borne pathogenic viruses. This shows that SAYAV evolved into a lineage independent of the known orthoflaviviruses. This study demonstrates a unique approach for understanding the epidemiology of uncultured viruses by combining viral metagenomics and pseudoinfectious viral particles.

Mpox Neutralizing Antibody Response to LC16m8 Vaccine in Healthy Adults.

BACKGROUND: Vaccination against mpox (formerly known as monkeypox), an infectious disease caused by the monkeypox virus (MPXV), is needed to prevent outbreaks and consequent public health concerns. The LC16m8 vaccine, a dried cell-cultured proliferative live attenuated vaccinia virus­based vaccine, was approved in Japan against smallpox and mpox. However, its immunogenicity and efficacy against MPXV have not been fully assessed. We assessed the safety and immunogenicity of LC16m8 against MPXV in healthy adults. METHODS: We conducted a single-arm study that included 50 participants who were followed up for 168 days postvaccination. The primary end point was the neutralizing antibody seroconversion rate against MPXVs, including the Zr599 and Liberia strains, on day 28. The secondary end points included the vaccine "take" (major cutaneous reaction) rate, neutralizing titer kinetics against MPXV and vaccinia virus (LC16m8) strains, and safety outcomes. RESULTS: Seroconversion rates on day 28 were 72% (36 of 50), 70% (35 of 50), and 88% (44 of 50) against the Zr599 strain, the Liberia strain, and LC16m8, respectively. On day 168, seroconversion rates decreased to 30% (15 of 50) against the Zr599 and Liberia strains and to 76% (38 of 50) against LC16m8. The vaccine "take" (broad definition) rate on day 14 was 94% (46 of 49). Adverse events (AEs), including common solicited cutaneous reactions, occurred in 98% (45 of 48) of participants; grade 3 severity AEs occurred in 16% (8 of 50). No deaths, serious AEs, or mpox onset incidences were observed up to day 168. CONCLUSIONS: The LC16m8 vaccine generated neutralizing antibody responses against MPXV in healthy adults. No serious safety concerns occurred with LC16m8 use. (Funded by the Ministry of Health, Labour and Welfare of Japan; Japan Registry of Clinical Trials number, jRCTs031220171.)

Cross-Neutralization Activities of Antibodies against 18 Lyssavirus Glycoproteins.

Some lyssaviruses, including the rabies virus (RABV), cause lethal neurological symptoms in humans. However, the efficacy of commercial vaccines has only been evaluated against RABV. To assess cross-reactivity among lyssaviruses, including RABV, sera from rabbits inoculated with human and animal RABV vaccines and polyclonal antibodies from rabbits immunized with expression plasmids of the glycoproteins of all 18 lyssaviruses were prepared, and cross-reactivity was evaluated via virus-neutralization tests using Duvenhage lyssavirus (DUVV), European bat lyssavirus-1 (EBLV-1), Mokola lyssavirus (MOKV), Lagos bat lyssavirus (LBV), and RABV. The sera from rabbits inoculated with RABV vaccines showed cross-reactivity with EBLV-1 and DUVV, both belonging to phylogroup I. However, reactivity with MOKV and LBV in phylogroup II was notably limited or below the detection level. Next, we compared the cross-reactivity of the polyclonal antibodies against all lyssavirus glycoproteins. Polyclonal antibodies had high virus-neutralization titers against the same phylogroup but not different phylogroups. Our findings indicate that a new vaccine should be developed for pre- and post-exposure prophylaxis against lyssaviral infections.

Establishment of serological neutralizing tests using pseudotyped viruses for comprehensive detection of antibodies against all 18 lyssaviruses.

Rabies is a fatal zoonotic, neurological disease caused by rabies lyssavirus (RABV) and other lyssaviruses. In this study, we established novel serological neutralizing tests (NT) based on vesicular stomatitis virus pseudotypes possessing all 18 known lyssavirus glycoproteins. Applying this system to comparative NT against rabbit sera immunized with current RABV vaccines, we showed that the current RABV vaccines fail to elicit sufficient neutralizing antibodies against lyssaviruses other than to those in phylogroup I. Furthermore, comparative NT against rabbit antisera for 18 lyssavirus glycoproteins showed glycoproteins of some lyssaviruses elicited neutralizing antibodies against a broad range of lyssaviruses. This novel testing system will be useful to comprehensively detect antibodies against lyssaviruses and evaluate their cross-reactivities for developing a future broad-protective vaccine.

High Seroprevalence of Severe Fever with Thrombocytopenia Syndrome Virus Infection among the Dog Population in Thailand.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne zoonotic disease caused by the SFTS virus (SFTSV). In Thailand, three human cases of SFTS were reported in 2019 and 2020, but there was no report of SFTSV infection in animals. Our study revealed that at least 16.6% of dogs in Thailand were seropositive for SFTSV infection, and the SFTSV-positive dogs were found in several districts in Thailand. Additionally, more than 70% of the serum samples collected at one shelter possessed virus-neutralization antibodies against SFTSV and the near-complete genome sequences of the SFTSV were determined from one dog in the shelter. The dog SFTSV was genetically close to those from Thailand and Chinese patients and belonged to genotype J3. These results indicated that SFTSV has already spread among animals in Thailand.

Pet Animals Were Infected with SARS-CoV-2 from Their Owners Who Developed COVID-19: Case Series Study.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection among pets owned by coronavirus disease 2019 (COVID-19) patients has been reported around the world. However, how often the animals are exposed to SARS-CoV-2 by their owners is still unclear. We have collected swab samples from COVID-19 patients' pets and performed real-time RT-PCR to detect the viral genome. In total, 8 of 53 dogs (15.1%) and 5 of 34 cats (14.7%) tested positive for the SARS-CoV-2 N gene. The result of a virus neutralization (VN) test also showed VN antibodies in four cats and six dogs. Our results indicate that the virus often passed from infected owners to their pets, which then excreted the virus despite having no or mild clinical signs.

Sociodemographic Factors, Mental Health, and Emotional Eating Associated With Concern for COVID-19 Contagion in Nurses: A Cross-Sectional Study.

OBJECTIVE: To determine the association between sociodemographic factors, stress, depression and anxiety, emotional eating, and concern about COVID-19 contagion in first and second-line Peruvian nurses. METHODOLOGY: The study was cross-sectional and conducted with 500 Peruvian nurses of both sexes, over 18 years of age. Validated questionnaires were used to measure sociodemographic aspects, concern about COVID-19, generalized anxiety, depression, self-perceived stress, and emotional eating. Multiple linear regression analysis was performed to analyze the factors affecting concern about COVID-19 contagion. RESULTS: The multiple linear regression analysis showed that stress, being between the ages of 18 and 29 years, being male, being from the coastal region or the jungle region, having a bachelor's degree, severe anxiety, and severe depression were associated with higher concern about COVID-19. On the other hand, having more than 5 to 10 years of experience and more than 10 years of experience, low emotional eating, and non-emotional eating were negatively associated with concern. This model explained 44.05% of the variability among the participating nurses. CONCLUSION: These findings provide resources for future research on the comprehensive well-being of nursing staff by exploring various sociodemographic aspects and mental conditions associated with greater concern about COVID-19. Meanwhile, years of experience and emotional eating behavior were associated with lower concern about COVID-19. Future studies could incorporate this information to preserve the mental and physical health of nurses in the face of potential occupational threats.

Detection of Mycobacterium tuberculosis transrenal DNA in urine samples among adult patients in Peru.

Diagnosis of tuberculosis (TB) relies on a sputum sample, which cannot be obtained from all symptomatic patients. Mycobacterium tuberculosis (Mtb) transrenal DNA (trDNA) has been detected in urine, an easily obtainable, noninvasive, alternative sample type. However, reported sensitivities have been variable and likely depend on collection/assay procedures and aspects of trDNA biology. We analyzed three serial urine samples from each of 75 adults with culture-confirmed pulmonary TB disease in Lima, Peru for detection of trDNA using short-fragment real-time PCR. Additionally, we examined host, urine, and sampling factors associated with detection. Overall sample sensitivity was 38% (95% Confidence Interval [CI] 30-45%). On a patient level (i.e., any of three samples positive), sensitivity was 73% (95% CI: 62-83%). Sensitivity was highest among samples from patients with smear-positive TB, 92% (95% CI: 62-100%). Specificity from a single sample from each of 10 healthy controls was 100% (95% CI: 69-100%). Adjusting our assay positivity threshold increased patient-level sensitivity to 88% (95% CI: 78-94%) overall without affecting the specificity. We did not find associations between Mtb trDNA detection and either patient characteristics or urine sample characteristics. Overall, our results support the potential of trDNA detection for TB diagnosis.

Low Replication Efficiency of a Japanese Rabbit Hepatitis E Virus Strain in the Human Hepatocarcinoma Cell Line PLC/PRF/5.

A Japanese rabbit hepatitis E virus (HEV) strain, JP-59, has been identified in a feral rabbit. When this virus was transmitted to a Japanese white rabbit, it caused persistent HEV infection. The JP-59 strain shares an <87.5% nucleotide sequence identity with other rabbit HEV strains. Herein, to isolate JP-59 by cell culture, we used a 10% stool suspension recovered from a JP-59-infected Japanese white rabbit and contained 1.1 × 107 copies/mL of the viral RNA and using it to infect a human hepatocarcinoma cell line, PLC/PRF/5. No sign of virus replication was observed. Although long-term virus replication was observed in PLC/PRF/5 cells inoculated with the concentrated and purified JP-59 containing a high titer of viral RNA (5.1 × 108 copies/mL), the viral RNA of JP-59c that was recovered from the cell culture supernatants was <7.1 × 104 copies/mL during the experiment. The JP-59c strain did not infect PLC/PRF/5 cells, but its intravenous inoculation caused persistent infection in rabbits. The nucleotide sequence analyses of the virus genomes demonstrated that a total of 18 nucleotide changes accompanying three amino acid mutations occurred in the strain JP-59c compared to the original strain JP-59. These results indicate that a high viral RNA titer was required for JP-59 to infect PLC/PRF/5 cells, but its replication capability was extremely low. In addition, the ability of rabbit HEVs to multiply in PLC/PRF/5 cells varied depending on the rabbit HEV strains. The investigations of cell lines that are broadly susceptible to rabbit HEV and that allow the efficient propagation of the virus are thus needed.

Subjective socioeconomic status: an alternative to objective socioeconomic status.

BACKGROUND: Subjective "ladder" measurements of socio-economic status (SES) are easy-to-administer tools that ask respondents to rate their own SES, allowing them to evaluate their own material resources and determine where it places them relative to their community. Here, we sought to compare the MacArthur Scale of Subjective Social status to the WAMI, an objective measure of SES that includes data on water and sanitation, asset ownership, education, and income. METHODS: Leveraging a study of 595 tuberculosis patients in Lima, Peru, we compared the MacArthur ladder score to the WAMI score using weighted Kappa scores and Spearman's rank correlation coefficient. We identified outliers that fell outside the 95th percentile and assessed the durability of the inconsistencies between scores by re-testing a subset of participants. We then used Akaike information criterion (AIC) to compare the predictability of logistic regression models evaluating the association between the two SES scoring systems and history of asthma. RESULTS: The correlation coefficient between the MacArthur ladder and WAMI scores was 0.37 and the weighted Kappa was 0.26. The correlation coefficients differed by less than 0.04 and the Kappa ranged from 0.26 to 0.34, indicating fair agreement. When we replaced the initial MacArthur ladder scores with retest scores, the number of individuals with disagreements between the two scores decreased from 21 to 10 and the correlation coefficient and weighted Kappa both increased by at least 0.03. Lastly, we found that when we categorized WAMI and MacArthur ladder scores into three groups, both had a linear trend association with history of asthma with effect sizes and AICs that differed by less than 15% and 2 points, respectively. CONCLUSION: Our findings demonstrated fair agreement between the MacArthur ladder and WAMI scores. The agreement between the two SES measurements increased when they were further categorized into 3-5 categories, the form in which SES is often used in epidemiologic studies. The MacArthur score also performed similarly to WAMI in predicting a socio-economically sensitive health outcome. Researchers should consider subjective SES tools as an alternative method for measuring SES, particularly in large health studies where data collection is a burden.

Simple and rapid detection of severe fever with thrombocytopenia syndrome virus in cats by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay using a dried reagent.

Severe fever with thrombocytopenia syndrome virus (SFTSV) causes lethal hemorrhagic diseases in human, cats, and dogs. Several human cases involving direct transmission of SFTSV from diseased animals have been reported. Therefore, rapid diagnosis in veterinary clinics is important for preventing animal-to-human transmission. Previously, we developed a simplified reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for human that does not require RNA extraction for detecting the SFTSV genome. In this study, we improved the simplified RT-LAMP assay for cats by introducing a dried reaction reagent and investigated the applicability of this method for diagnosing SFTS in cats. SFTSV RNA was detected in 11 of 12 cats naturally infected with SFTSV by RT-LAMP assay using both liquid and dried reagents. The RT-LAMP assay using liquid and dried reagents was also applicable to the detection of SFTSV genes 3-4 days after challenge in cats experimentally infected with SFTSV. The minimum copy number of SFTSV genes for 100% detection using the RT-LAMP assay with liquid and dried reagents was 4.3 × 104 and 9.6 × 104 copies/mL, respectively. Although the RT-LAMP assay using the dried reagent was less sensitive than that using the liquid reagent, it was sufficiently sensitive to detect SFTSV genes in cats with acute-phase SFTS. As the simplified RT-LAMP assay using a dried reagent enables detection of SFTSV genes more readily than the assay using a liquid reagent, it is applicable for use in veterinary clinics.

Lethal Disease in Dogs Naturally Infected with Severe Fever with Thrombocytopenia Syndrome Virus.

Severe fever with the thrombocytopenia syndrome virus (SFTSV) causes fatal disease in humans, cats, and cheetahs. In this study, the information on seven dogs with SFTS was summarized. All dogs showed anorexia, high fever, leukopenia, and thrombocytopenia, two dogs showed vomiting and loose stool, and five dogs had tick parasites. All dogs also had a history of outdoor activity. The SFTSV gene was detected in all dogs. Remarkably, three dogs (43%) died. SFTSV was isolated from six dogs and the complete genomes were determined. A significant increase in anti-SFTSV-IgG antibodies was observed in two dogs after recovery, and anti-SFTSV-IgM antibodies were detected in four dogs in the acute phase. Using an ELISA cut-off value of 0.410 to discriminate between SFTSV-negative and positive dogs, the detection of anti-SFTSV-IgM antibodies was useful for the diagnosis of dogs with acute-phase SFTS. Four out of the ninety-eight SFTSV-negative dogs possessed high anti-SFTSV IgG antibody titers, indicating that some dogs can recover from SFTSV infection. In conclusion, SFTSV is lethal in some dogs, but many dogs recover from SFTSV infection.

Risk assessment of infection with severe fever with thrombocytopenia syndrome virus based on a 10-year serosurveillance in Yamaguchi Prefecture.

In Japan, the first patient with severe fever with thrombocytopenia syndrome was reported in Yamaguchi in 2012. To understand the severe fever with thrombocytopenia syndrome virus (SFTSV) infection in this region, a retrospective surveillance in sika deer and wild boars in Yamaguchi was conducted using a virus-neutralizing (VN) test. The result revealed that 510 of the 789 sika deer and 199 of the 517 wild boars were positive for anti-SFTSV antibodies. Interestingly, seroprevalence in sika deer increased significantly from 2010-2013 to 2015-2020. The SFTSV gene was detected in one of the 229 serum samples collected from sika deer, but not from wild boars. In conclusion, SFTSV had spread among wild animals before 2012 and expanded gradually around 2013-2015 in Yamaguchi.

Roles of raccoons in the transmission cycle of severe fever with thrombocytopenia syndrome virus.

The present study investigated severe fever with thrombocytopenia syndrome virus (SFTSV) infection in raccoons in Wakayama Prefecture from 2007 to 2019. To perform surveillance, an enzyme-linked immunosorbent assay (ELISA) was established, and the sensitivity and specificity of the ELISA were 100% in comparison with a 50% focus-reduction neutralization assay. Using the established ELISA, we performed serosurveillance of SFTSV infection in 2,299 raccoons in Tanabe region, Wakayama Prefecture from 2007 to 2019. The first anti-SFTSV-positive raccoon was captured in October 2009. The seroprevalence of SFTSV infection was <10% between April 2009 and March 2013, 23.9% between April 2013 and March 2014, 37.5% between April, 2014 and March 2015, and over 50% from April 2015. Next, we performed detection of SFTSV genes in sera of raccoons captured in Wakayama Prefecture after April 2013. The results indicated that 2.4% of raccoons were positive for SFTSV genes and that the frequency of SFTSV infection among raccoons between January and March (0.7%) was lower than that between April and June (3.4%). In addition, virus genes were detected from many specimens, including sera and feces of two raccoons, and viral antigens were detected in lymphoid cells in lymphoid follicles in the colon by immunohistochemical staining. In conclusion, SFTSV had recently invaded the area and had rapidly spread among wild animals. The first patient in this area was reported in June 2014, indicating that raccoons are good sentinels for assessing the risk of SFTSV in humans.

Nationwide survey of hepatitis E virus infection among wildlife in Japan.

In Japan, hepatitis E virus (HEV) causes hepatitis in humans through the consumption of raw or undercooked meat, including game meat. In the present study, nationwide surveillance of HEV infection among a total of 5,557 wild animals, including 15 species, was conducted in Japan. The prevalence of anti-HEV antibodies in wild boar was 12.4%, with higher positive rates in big boars (over 50 kg, 18.4%) than in small individuals (less than 30 kg, 5.3%). Furthermore, HEV RNA was more frequently detected in piglets than in older boars. Interestingly, the detection of HEV among wildlife by ELISA and RT-PCR suggested that HEV infection in Sika deer was a very rare event, and that there was no HEV infection among wild animals except for wild boar, Sika deer and Japanese monkeys. In conclusion, wild boar, especially piglets, are at high risk of HEV infection, while other wild animals showed less risk or no risk of HEV transmission.

Zoonotic Infection with Oz Virus, a Novel Thogotovirus.

Oz virus is a novel thogotovirus isolated from ticks that causes lethal infection in mice. We conducted serosurveillance of Oz virus infection among humans and wild mammals in Japan using virus-neutralization tests and ELISAs. Results showed that Oz virus may be naturally infecting humans and other mammalian hosts.

A serological survey and characterization of Getah virus in domestic pigs in Thailand, 2017-2018.

Getah virus (GETV) is a mosquito-borne RNA virus belonging to the family Togaviridae, genus Alphavirus. GETV infection causes diarrhoea and death in piglets, and reproductive failure and abortion in sows. This study conducted a serological survey of GETV infection among domestic pig populations in Thailand. ELISA was used to analyse 1,188 pig serum samples collected from 11 provinces of Thailand during 2017-2018, with 23.1% of the samples being positive for anti-GETV antibodies. The positive ratio of anti-GETV antibodies was significantly higher in nursery (67.9%) and older stages (84.5%) of pigs than in finishing stage (14.2%). Furthermore, we successfully isolated GETV from one pig serum, designated as GETV strain GETV/SW/Thailand/2017, and determined the complete genome sequence (11,689 nt). Phylogenetic analysis demonstrated that our isolate was different from the recent GETV group spreading among pig populations in East Asia and formed a cluster with two GETV strains, namely YN12031 (China, 2015) and LEIV16275Mar (Far-East Russia, 2007). We concluded that two different GETV groups are currently spreading among pig populations in Asian countries.

Epidemiological study of Kabuto Mountain virus, a novel uukuvirus, in Japan.

Kabuto Mountain virus (KAMV), the new member of the genus Uukuvirus, was isolated from the tick Haemaphysalis flava in 2018 in Japan. To date, there is no information on KAMV infection in human and animals. Therefore, serological surveillance of the infection among humans and wild mammals was conducted by virus-neutralization (VN) test and indirect immunofluorescence assay (IFA). Sera of 24 humans, 59 monkeys, 171 wild boars, 233 Sika deer, 7 bears, and 27 nutria in Yamaguchi Prefecture were analyzed by VN test. The positive ratio of humans, monkeys, wild boars, and Sika deer were 20.8%, 3.4%, 33.9% and 4.7%, respectively. No positive samples were detected in bears and nutria. The correlation coefficients between VN test and IFA in human, monkey, wild boar, and Sika deer sera were 0.5745, 0.7198, 0.9967 and 0.9525, respectively. In addition, KAMV was detected in one pool of Haemaphysalis formosensis ticks in Wakayama Prefecture. These results indicated that KAMV or KAMV-like virus is circulating among many wildlife and ticks, and that this virus incidentally infects humans.

Regeneración ósea en defectos calvariales de ratones NOD. CB17- Prkdcscid/J (NOD SCID) a partir de células madre de la pulpa dental: una revisión sistemática / Bone regeneration in calvarial defects of NOD. CB17- Prkdcscid/J (NOD SCID) mice from dental pulp stem cells: a systematic review

Resumen Introducción: las células madre de la pulpa dental humana presentan una capacidad osteogénica muy significativa, lo que permite la formación de hueso nuevo que se adapta muy bien al existente. El objetivo fue evaluar la eficacia de la regeneración ósea a partir de células madre de la pulpa dental, analizando estudios in vivo realizados en ratones NOD.CB17-Prkdcscid/J (NOD SCID) inmunocompetentes. Métodos: se analizaron estudios entre el 2016 al 2020, encontrados en Pubmed, ScienceDirect y Lilacs. Para realizar la revisión sistemática se siguió las directrices PRISMA, la evaluación de la calidad y del riesgo de sesgo se realizó considerando los criterios expuestos en la herramienta National Heart Lung and Blood Institute - NHLBI. Resultados: aplicando los criterios de inclusión y exclusión, se seleccionaron 6 investigaciones las cuales se evaluaron, eligiéndose solo 2 de ellas para su revisión (n=1859). Los datos de los estudios se extrajeron y ordenaron obedeciendo los detalles del estudio, metodología del análisis y resultados. Conclusión: los resultados obtenidos demuestran que existe una regeneración ósea por parte de las células madre derivadas de la pulpa dental, debido a que promueven la osteogénesis y el hueso nuevo se une satisfactoriamente al hueso existente. El análisis in vitro indica que es una excelente fuente osteogénica y los estudios in vivo corroboran que los medios de cultivo interfieren en la formación de hueso. Se requieren de más estudios para afianzar la formación de hueso por parte de las Células Madre de la Pulpa Dental - DPSC.

Abstract Introduction: human dental pulp stem cells show a very significant osteogenic capacity, allowing the formation of new bone that is very well adapted to existing bone. The objective was to evaluate the efficacy of bone regeneration from dental pulp stem cells, analyzing in vivo studies carried out in immunocompetent NOD. CB17-Prkdcscid/J (NOD SCID) mice. Methods: we analysed studies from 2016 to 2020, found in Pubmed, ScienceDirect and Lilacs. The systematic review followed the PRISMA guidelines, the quality and risk of bias assessment was performed considering the criteria set out in the National Heart Lung and Blood Institute - NHLBI tool. Results: applying the inclusion and exclusion criteria, 6 studies were selected and evaluated, and only 2 of them were selected for review (n=1859). Data from the studies were extracted and sorted according to study details, analysis methodology and results. Conclusion: the results obtained show that there is bone regeneration by stem cells derived from dental pulp, because they promote osteogenesis and the new bone successfully joins the existing bone. In vitro analysis indicates that it is an excellent osteogenic source and in vivo studies confirm that culture media interfere with bone formation. Further studies are required to confirm bone formation by Dental Pulp Stem Cell - DPSC.

Characterization of rabbit hepatitis E virus isolated from a feral rabbit.

Rabbit hepatitis E virus (HEV) has been detected among rabbits and recently isolated from immunocompromised patients, suggesting zoonotic transmission. In this study, HEV infection among feral rabbits (Oryctolagus cuniculus) was assessed by detection of anti-HEV antibodies and HEV RNA. The prevalence of anti-HEV antibodies in sera was of 33 % (20/60) and HEV RNA was detected from only one of fecal swabs (1.7 %, 1/58). Furthermore, one naïve rabbit was intravenously inoculated with the suspension of the HEV-positive fecal specimen, exhibiting persistent HEV shedding in feces, intermittent viremia, seroconversion to anti-HEV IgM and IgG, and high alanine aminotransferase (ALT) values, indicating persistent HEV infection. The isolate JP-59 had a length of 7,282 bp excluding a poly (A) tail and possessed the characteristic 93 bp-insertion in ORF1. Phylogenetic analysis indicated that JP-59 formed a cluster with other rabbit HEV isolates from rabbits and human origin. The JP-59 shared the nucleotide sequence identities less than 87 % with other rabbit HEVs, suggesting that a novel rabbit HEV strain was circulating in Japan.