Since 2017, hepatitis A virus (HAV) infection has been an epidemic among men who have sex with men (MSM) in Japan. We have come across 11 MSM patients with hepatitis A who were also infected with HIV. In 1999–2000, we came across 5 HIV-infected patients with hepatitis A. Since the conditions of current HIV-infected patients have changed owing to the recent progress in anti-HIV therapies, we compared clinical features of hepatitis A between patients in 2017–2018 and those in 1999–2000. By comparing the background characteristics of the patients, we found that the CD4/CD8 ratio was significantly higher in the 2017–2018 group. After the onset of hepatitis, peak levels of hepatic transaminases were found to be higher in the 2017–2018 group, suggesting severe hepatocellular damage. In contrast, neither the peak level of total bilirubin nor the nadir of prothrombin time was significantly different among the 2 groups. We also analyzed the HAV genome derived from some of the recently infected patients, and found that the HAV strains were almost the same among these patients; slight differences were observed from the previously identified strain. Thus, we concluded that the recovery of immunity by recent anti-HIV therapies may result in more severe hepatocellular damages and differences in clinical features.

The estimation of prevalence and genotype distribution of high-risk human papillomavirus (HR-HPV) is important in formulating strategies for the prevention and screening of cervical cancer. This study aimed to determine HR-HPV prevalence and genotype distribution among Chinese women living in Sichuan Province. A total of 3,089 women were recruited from the Third People’s Hospital of Chengdu in Sichuan Province. HR-HPV genotyping was assessed using real-time quantitative polymerase chain reaction. Up to 720 women (23.3%) were HR-HPV positive. A total of 81.1% (584/720) of women had a single HR-HPV genotype, and 18.9% (136/720) had multiple genotypes. The most frequently detected HPV genotype was HPV52, followed by HPV58, HPV16, and HPV51. Age subgroup analysis showed 2 peaks for the frequencies of HPV infections, one for the group of women under 29 years old, and the other for the group of women over 50 years old. Among the women for whom cytology results were available, HPV16 was the most commonly detected genotype among women affected with high-grade squamous intraepithelial lesion (32.5%) and squamous cell carcinoma, followed by HPV58, HPV33, and HPV52. This is one of the largest studies to demonstrate HPV prevalence and genotype distribution among Chinese women in Sichuan. The prevalence and genotype distribution of HR-HPV was unique with higher frequencies of HPV52 and HPV58.

The annual number of outbreaks of Vibrio parahaemolyticus and Salmonella food poisoning and that of patients in Japan, from 2000 to 2018, decreased exponentially even though the size of the individual outbreaks (the number of patients per outbreak) tended to become larger. For food poisonings caused by Campylobacter, the annual number of outbreaks increased exponentially while outbreak size became smaller and the annual number of patients remained almost unchanged. For food poisoning caused by norovirus, both the number of outbreaks and that of patients remained high throughout. Over time, the geographical and seasonal distribution of food poisonings became narrower for Vibrio parahaemolyticus and Salmonella, while they became wider for Campylobacter and norovirus. Further analyses using the attack rate-patient number plots suggested that the number of the outbreaks was determined mainly by the levels of microbial contamination of foods before they were brought into the facilities for consumption.

Fever is a systemic inflammatory response of the body to pyrogens. Nuclear factor κB (NF-κB) is a central signaling molecule that causes the excessive secretion of various pyrogen-induced pro-inflammatory factors. This study explored the feasibility of a novel reporter gene assay (RGA) for pyrogen detection using RAW264.7 cells stably transfected with the NF-κB reporter gene as a pyrogenic marker. The RGA could detect different types of pyrogens, including the lipopolysaccharide of gram-negative bacteria, the lipoteichoic acid of gram-positive bacteria, and the zymosan of fungi, and a good dose-effect relationship was observed in terms of NF-κB activity. The limits of detection of the RGA to those pyrogens were 0.03 EU/ml, 0.001 μg/ml, and 1 μg/ml, respectively. The method had good precision and accuracy and could be applied to many molecules (e.g., nivolumab, rituximab, bevacizumab, etanercept, basiliximab, Haemophilus influenzae type b conjugate vaccine, 23-valent pneumococcal polysaccharide vaccine, group A and group C meningococcal conjugate vaccine, diphtheria, tetanus, pertussis [acellular, component], poliomyelitis [inactivated] vaccine, and imject alum adjuvant). The results of this study suggest that the novel RGA has a wide pyrogen detection spectrum and is sufficiently sensitive, stable, and accurate for various applications.

Non-typhoidal salmonellae (NTS) are a major cause of acute diarrhea with characteristic multidrug resistance (MDR). In a hospital-based study, 81 NTS were isolated and tested for serotypes and antimicrobial resistance (AMR). Salmonella enterica isolates were classified into 7 different typable serovars, however, 19 (23%) isolates remained untypable. The most common serovars were S. Kentucky (48%), and S. Virchow (22%). Most of the NTS isolates displayed resistance to nalidixic acid (NA) (73%), ciprofloxacin (CIP) (48%), ampicillin (AM) and norfloxacin (NOR) (36% each), and gentamicin (CN) (31%). The AMR profiles for CN and NA; and AM, CIP, NA and NOR, were found to be high in S. Virchow (83%) and S. Kentucky (43%), respectively. Analysis of the pulsed-field gel electrophoresis patterns of S. Kentucky revealed 3 clusters. S. Kentucky has clones closely related to become prominent in recent years in Delhi. The AMR appears to be consistent with the change in MDR patterns during 2014-2017. The observed prevalence of S. Kentucky and S. Virchow in large numbers of diarrheal cases is novel. The NTS are mostly resistant to fluoroquinolones, which is the current drug of choice for treating diarrheal cases. MDR is very common among clonally related S. Kentucky.

Long-lasting insecticidal nets (LLINs) experience some operational problems that reduce their effectiveness, such as limited spaces for hanging, biting of mosquitoes outdoors, a shift of key biting time from midnight to dawn or dusk, and development of pyrethroid resistance in mosquitoes. The concept of spatial repellency may be a countermeasure to overcome the above issues. The effect of the combined use of metofluthrin-impregnated spatial repellent devices (MSRDs) and LLINs (Olyset^® Plus) on malaria prevalence and vector mosquitoes were examined in malaria endemic villages in south-eastern Malawi. The intervention reduced the infection rate in children as well as the number of pyrethroid-resistant vector mosquitoes. To achieve effective malaria control, continued intervention using MSRDs with 2 strips per 10 m² at 3-month intervals to reduce the density of malaria mosquitoes is recommended.

Trichodysplasia spinulosa-associated polyomavirus (TSPyV or human polyomavirus 8) was identified from patients with trichodysplasia spinulosa, a rare skin disease affecting the faces of immunocompromised patients. Like other polyomaviruses, the TSPyV genome encodes a large T antigen (LT). However, the expression and functions of TSPyV LT in infected cells remain largely unknown. In the present study, we cloned a full-length TSPyV LT cDNA from cells transfected with the full-length of TSPyV LT DNA. Transfection study using green fluorescence protein-tagged LT expression plasmids showed that TSPyV LT was expressed in the nucleus of transfected cells. Analysis of deletion mutants identified a nuclear localization signal in TSPyV LT. Recombinant TSPyV LT exhibited an ATPase activity. TSPyV LT has a chitinase-like domain; however, no chitinase activity was detected. Immunoprecipitation assays revealed that TSPyV LT bound to retinoblastoma 1, but not to p53 in transfected cells. Expression of TSPyV LT in NIH3T3 cells induced colony formation in soft agar, suggesting its transformation activity. These data indicate that TSPyV LT may be associated with the pathogenesis of trichodysplasia spinulosa, which is a hyperplasia of keratinocytes in inner hair follicles.

Dengue is one of the most serious mosquito-borne viral diseases occurring in humans. To combat the complexity of 4 antigenically distinct serotypes, the ideal vaccine for dengue should be able to stimulate cross-neutralizing antibodies. Recently, genetics-based immune responses have been studied to guide vaccine design against several viral pathogens. Despite a recent approval of dengue vaccine, information on genetics-based immune responses against dengue virus (DENV) is still limited. Consequently, we aimed to determine the profiles of immunoglobulin heavy chain genes from DENV2 infected patients. The immunoglobulin heavy chain variable region genes (IGHV) were amplified from peripheral blood mononuclear cells of DENV2 secondary infected patients in the acute, convalescence, and recovery phases. Antibody heavy chain genes were sequenced using next-generation sequencing, and analyzed to identify correlations with neutralizing and enhancing activities of the serum samples. IGHV1-69, 3-23, and 3-30 were frequently discovered in our Thai DENV2 infected patients. Our findings provide new data on the human B cell response during secondary DENV2 infections in Thai dengue patients that offer supportive information for dengue vaccine design and therapeutics development.

Early-onset sepsis (EOS) in neonates is a serious disease with severe complications. The increased severity of EOS and risk of death in newborns in recent years signify that continued monitoring to detect possible changes in the pathogen etiology, disease severity, and disease outcome is particularly important. We conducted a retrospective study on early-onset infection among infants (birth weights > 800 g) who were hospitalized in the Children’s Hospital of Soochow University from January 1, 2011, to December 31, 2017. Multivariable analysis was performed to determine the significant predictors of mortality. The most frequent early-onset pathogen was Group B Streptococcus (GBS) (28.1%), followed by Escherichia coli (21.6%), Listeria monocytogenes (11.8%), and Klebsiella pneumoniae (7.8%). Most infants (85.6%) with early-onset infections survived until hospital discharge, while 44 (14.4%) patients died. Multivariable logistic regression analysis showed that the significant predictors of mortality were the pathogen (GBS, E. coli, or other pathogens) and birth weight (both P < 0.01). GBS remains the most frequent pathogen known to infect infants. E coli was the most common pathogen associated with neonatal mortality. Prevention of E. coli sepsis, specifically among preterm infants, remains a challenge.

Zika is a mosquito-borne disease that has been posing a significant threat to public health in recent years. The Zika virus (ZIKV), the causative agent of this disease, is classified into 2 distinct genetic lineages, namely Asian and African. While molecular nucleic acid analysis methods have been shown to be useful for the diagnosis of ZIKV infection, the development of assays based on one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) offers several advantages, such as shorter incubation times, ease of handling, and rapid detection. In this study, a universal LAMP primer set was developed to target conserved sequences of known ZIKV lineages. Additionally, the Af7462 and As1788 primer sets were designed based on LAMP-based single-nucleotide polymorphism (SNPs) typing for the specific detection of the African and Asian lineages. The developed RT-LAMP assays could specifically detect the African and Asian lineages of ZIKV, with a detection limit ranging from 0.17 FFU/mL to 2.3×10² FFU/mL. As ZIKV viremia ranges between 10² to 10⁶ PFU/mL or 10³ to 10⁶ copies/mL, the data indicate that the viremia range of clinical samples is within the detection range of our assay. Due to the high specificity and sensitivity, as well as the ease of use of our assay, it could potentially be used for early clinical diagnosis applications.

Approximately 40 genotypes of the human papillomavirus (HPV) have been identified in cervical mucosa. In particular, HPV-16 and HPV-18 have been associated with cervical neoplasia. Squamous intraepithelial lesions (SILs) are precursors of cervical cancer. This study aimed to identify the HPV by genotype in SILs using a linear array genotyping test in a population in Mexico. We performed a cross-sectional study of 129 female patients with or without SILs, as determined by colposcopy, who completed a risk factor questionnaire. Cervical swab samples were obtained and genotyped using a Linear Array HPV Genotyping assay. Forty-nine (37.98%) samples were positive for HPV, and 24 genotypes were identified among these samples. The most common genotype was HPV-16. Twelve genotypes were found in both high- and low-grade SILs (HPV-6, 16, 31, 39, 51, 52, 53, 58, 59, 61, 67, and 84), of which seven were high-risk SILs (HPV-16, 31, 39, 51, 52, 58, and 59). Among the populations studied, the most frequent genotype was HPV-16, multiple infections were found, and four patients without injury tested positive for HPV.

An epidemic of rotavirus (RV) gastroenteritis occurred from April to July 2015 across a wide area of Hokkaido, surrounding the Abashiri-Kosei General Hospital. The RV vaccine for children in Shari and Koshimizu was provided at public funds by their local governments, while children in Abashiri were charged for the vaccine. This study examined the effectiveness of the RV vaccine against the risk of hospitalization based on a retrospective cohort study and the impact of using public funds for RV vaccination on a regional RV gastroenteritis epidemic. The vaccination coverage was significantly higher in children in Shari and Koshimizu than in Abashiri (87.8% vs. 42.7%, respectively, p < 0.001). The RV gastroenteritis-related risk of hospitalization was slightly lower in children from Shari and Koshimizu than in those from Abashiri (1.6% vs. 3.2%, respectively, p = 0.07). In addition, the risk of hospitalization in the vaccinated children was significantly lower than that in the unvaccinated children (0.7% vs. 4.8%, respectively, p < 0.001); indicating that the RV vaccine effectiveness against the risk of hospitalization was 96.5% (95% confidence interval 45.7%–99.8%). In conclusion, the use of public funds for the provision of RV vaccine increased the vaccination coverage, which, in combination with high vaccine effectiveness, led to a decrease in the number of hospitalizations in children during a regional RV gastroenteritis epidemic.

Oropouche virus (OROV) is a negative-sense, single-stranded RNA arbovirus transmitted to humans by the midge Culicoides paraenesis, causing Oropouche fever. Reports of its outbreak in Brazil have so far been restricted to the Central-Northern region of the country. However, its incidence is underestimated, mainly due to its clinical similarities with other arbovirus diseases, including dengue (DENV), chikungunya (CHIKV), and zika (ZIKV), and the lack of specific diagnostic tests. Here, we report for the first time, the detection of OROV in saliva and urine samples, and cases of autochthone OROV infections in Salvador Metropolitan region, Bahia, a Northeastern capital in the coast of Brazil. Serum, saliva, and urine samples negative for DENV, CHIKV, and ZIKV were tested for OROV using a reverse transcription nested polymerase chain reaction (RT-nested-PCR) protocol, and 2 serum, 2 saliva, and 1 urine samples were positive. This report shows the need for an efficient surveillance system for controlling the spread of this virus, and suggests the use of saliva and urine as alternative samples for OROV detection in the absence of serum samples.

A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla_KPC, bla_IMP, bla_NDM, bla_VIM, bla_OXA-48-like, and bla_GES. Of 70 bla_IMP variants, 67 subtypes were simulated to be PCR-positive based on in silico simulation and the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

Volume 72 no 6, p.413–419, 2019. Page 418, Acknowledgments “We would like to thank all staff and members of the Department of Virology, NEKKEN, Nagasaki University, Japan for providing technical support and advice. Our special thanks to the staff of the Pavilion II and the Central Laboratory of San Lazaro Hospital for their kind assistance during patient recruitment and data collection. We are also very grateful for the support of the Senior Vice President and Head of Research and Biotechnology (R&B) Group of St. Luke’s Medical Center, Dr. Isaac David E. Ampil II. Finally, our sincere thanks to the members of R&B’s dengue research group for kindly preparing the samples to be transported to NEKKEN.” should read “This research was supported by grants from the Japan Agency for Medical Research and Development (AMED) under Grant Number JP18fm0108001, JP19fm0108001 (Japan Initiative for Global Research Network on Infectious Diseases (J-GRID)), AMED Research on Emerging and Re-emerging Infectious Diseases (19fk0108035j0003) and e-ASIA Joint Research Program and; Philippine Council for Health Research and Development (PCHRD) of the Department of Science and Technology (DOST), Philippines, with partial support from the Research and Biotechnology of St. Luke’s Medical Center (R&B-SLMC), Philippines (Project No. 07-024). Funders have no role in the study design, data collection, and interpretation, or the decision to submit this work for publication. We would like to thank all staff and members of the Department of Virology, Institute of Tropical Medicine (NEKKEN), Nagasaki University, Japan, for providing technical support and advice. Our special thanks to the staff of the Pavilion II and the Central Laboratory of San Lazaro Hospital for their kind assistance during patient recruitment and data collection. We are also very grateful for the support of the Senior Vice President and Head of Research and Biotechnology (R&B) Group of St. Luke’s Medical Center, Dr. Isaac David E. Ampil II. Finally, our sincere thanks to the members of R&B’s Dengue Research Group for kindly preparing the samples to be transported to NEKKEN.”