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Objective: To evaluate treatment patterns, healthcare resource utilization (HRU) and costs among peripheral T-cell lymphoma (PTCL) patients in the USA. Methods: A retrospective cohort study, using the IQVIA PharMetrics® Plus claims database from 1 April 2011 to 30 November 2021, identified PTCL patients receiving systemic treatments. Three mutually exclusive subcohorts were created based on line of therapy (LOT): 1LOT, 2LOT and ≥3LOT. Common treatment regimens, median time on treatment, all-cause and PTCL-related HRU and costs were estimated. Results: Among 189 PTCL patients identified, 61.9% had 1LOT, 21.7% had 2LOT and 16.4% had ≥3LOT. The most common treatment regimens in the 1LOT were CHOP/CHOP-like, CHOEP/CHOEP-like and brentuximab vedotin; monotherapies were most common in the 2LOT and ≥3LOT. All-cause and PTCL-related hospitalizations and prescriptions PPPM increased with increasing LOT. Nearly 70% of total treatment costs were PTCL related. Conclusion: Higher utilization of combination therapies in the 1LOT and monotherapies in subsequent LOTs were observed, alongside high PTCL-related costs.
Peripheral T-cell lymphomas (PTCL) are a rare and fast-growing form of blood cancer. About 800012,000 people in the USA are diagnosed with PTCL every year. As it is a rare disease and has many types, and there is a limited understanding of the patients who have PTCL and the treatments they receive in the real world. The purpose of this study was to evaluate how these patients are treated, what are they treated with and what are the costs of these treatments in the USA. The data collected on these patients was divided into three groups based upon the number of lines of treatment/therapy (LOT) they received: 1LOT, 2LOT and ≥3LOT. This study researched different treatments and their duration in each line of therapy. Among 189 PTCL patients included in the study, the average age of patients was 55 years and 62% were male. Among these patients, 62% had 1LOT, 22% had 2LOT and 16% had ≥3LOT. The most common treatments in the 1LOT were traditional chemotherapy regimens followed by targeted therapies: CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) or CHOP-like, CHOEP (cyclophosphamide, doxorubicin, vincristine, etoposide and prednisone) or CHOEP-like, and brentuximab vedotin. Treatment regimens with only one drug were most common in the 2LOT and ≥3LOT. The total cost of PTCL treatment in the USA is very high; 70% of this cost is related to their treatment with various drugs. More research is needed to better understand the treatment and cost of this rare cancer.
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Linfoma de Células T Periférico , Humanos , Linfoma de Células T Periférico/tratamiento farmacológico , Linfoma de Células T Periférico/epidemiología , Estudios Retrospectivos , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Brentuximab Vedotina/uso terapéutico , Costos de la Atención en Salud , Doxorrubicina , Ciclofosfamida/uso terapéutico , Vincristina/uso terapéutico , PrednisonaRESUMEN
Norovirus (NoV) genogroup II, polymerase type P31, capsid genotype 4, Sydney_2012 variant (GII.P31/GII.4_Sydney_2012) has been circulating at high levels for over a decade, raising the question of whether this strain is undergoing molecular alterations without demonstrating a substantial phylogenetic difference. Here, we applied next-generation sequencing to learn more about the genetic diversity of 14 GII.P31/GII.4_Sydney_2012 strains that caused epidemics in a specific region of Japan, with 12 from Kyoto and 2 from Shizuoka, between 2012 and 2022, with an emphasis on amino acid (aa) differences in all three ORFs. We found numerous notable aa alterations in antigenic locations in the capsid region (ORF2) as well as in other ORFs. In all three ORFs, earlier strains (2013-2016) remained phylogenetically distinct from later strains (2019-2022). This research is expected to shed light on the evolutionary properties of dominating GII.P31/GII.4_Sydney_2012 strains, which could provide useful information for viral diarrhea prevention and treatment.
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Evolución Molecular , Norovirus , Japón/epidemiología , Filogenia , Evolución Biológica , Proteínas de la Cápside/genética , Norovirus/genéticaRESUMEN
An increasing trend of sapovirus (SaV) infections in Japanese children during 2009-2019, particularly after the introduction of the voluntary rotavirus (RV)-vaccination program has been observed. Herein, we investigated the epidemiological situation of SaV infections from 2019 to 2022 when people adopted a precautionary lifestyle due to the emergence of the COVID-19 pandemic, and RV vaccines had been implemented as routine vaccines. Stool samples were collected from children who attended outpatient clinics with acute gastroenteritis and analyzed by reverse transcriptase-polymerase chain reaction to determine viral etiology. Among 961 stool samples, 80 (8.3%) were positive for SaV: 2019-2020 (6.5%), 2020-2021 (0%), and 2021-2022 (12.8%). The trend of SaV infection in Japanese children yet remained upward with statistical significance (p = 0.000). The major genotype was GI.1 (75%) which caused a large outbreak in Kyoto between December 2021 and February 2022. Phylogenetic, gene sequence and deduced amino acid sequence analyses suggested that these GI.1 strains detected in the outbreak and other places during 2021-2022 or 2019-2020 remained genetically identical and widely spread. This study reveals that SaV infection is increasing among Japanese children which is a grave concern and demands immediate attention to be paid before SaV attains a serious public health problem.
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COVID-19 , Infecciones por Caliciviridae , Sapovirus , Vacunas , Niño , Humanos , Sapovirus/genética , Japón/epidemiología , Filogenia , Pandemias , Heces , COVID-19/epidemiología , Genotipo , Infecciones por Caliciviridae/epidemiologíaRESUMEN
Noroviruses (NoVs) are a global concern, causing widespread outbreaks and sporadic acute gastroenteritis (AGE) cases across all age groups. Recent research has shed light on the emergence of novel recombinant strains of NoV in various countries. To delve deeper into this phenomenon, we extensively analyzed 1,175 stool samples collected from Japanese infants and children with AGE from six different prefectures in Japan over three years, from July 2018 to June 2021. Our investigation aimed to determine the prevalence and genetic characteristics of NoV associated with sporadic AGE while exploring the possibility of detecting NoV recombination events. Among the analyzed samples, we identified 355 cases positive for NoV, 11 cases attributed to GI genotypes, and 344 associated with GII genotypes. Notably, we discovered four distinct GI genotypes (GI.2, GI.3, GI.4, and GI.6) and seven diverse GII genotypes (GII.2, GII.3, GII.4, GII.6, GII.7, GII.14, and GII.17). The predominant genotypes were GII.4 (56.4%; 194 out of 344), followed by GII.2 and GII.3. Through dual genotyping based on sequencing of the ORF1/ORF2 junction region, we identified a total of 14 different RdRp/capsid genotypes. Of particular interest were the prevalent recombinant genotypes GII.4[P31] and GII.2[P16]. Notably, our study revealed a decrease in the number of children infected with NoV during and after the COVID-19 pandemic. These findings underscore the importance of continuous NoV surveillance efforts.
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Infecciones por Caliciviridae , Variación Genética , Norovirus , Niño , Preescolar , Humanos , Lactante , Infecciones por Caliciviridae/epidemiología , Infecciones por Caliciviridae/virología , COVID-19 , Heces/virología , Genotipo , Japón/epidemiología , Norovirus/clasificación , Norovirus/genética , Filogenia , Prevalencia , Adolescente , Proteínas de la Cápside/genéticaRESUMEN
Transforming growth factor-beta 1 (TGF-ß1) is a pleiotropic growth factor playing various roles in the human body including cell growth and development. More functions of TGF-ß1 have been discovered, especially its roles in viral infection. TGF-ß1 is abundant at the maternal-fetal interface during pregnancy and plays an important function in immune tolerance, an essential key factor for pregnancy success. It plays some critical roles in viral infection in pregnancy, such as its effects on the infection and replication of human cytomegalovirus in syncytiotrophoblasts. Interestingly, its role in the enhancement of Zika virus (ZIKV) infection and replication in first-trimester trophoblasts has recently been reported. The above up-to-date findings have opened one of the promising approaches to studying the mechanisms of viral infection during pregnancy with links to corresponding congenital syndromes. In this article, we review our current and recent advances in understanding the roles of TGF-ß1 in viral infection. Our discussion focuses on viral infection during pregnancy, especially in the first trimester. We highlight the mutual roles of viral infection and TGF-ß1 in specific contexts and possible functions of the Smad pathway in viral infection, with a special note on ZIKV infection. In addition, we discuss promising approaches to performing further studies on this topic.
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Infección por el Virus Zika , Virus Zika , Embarazo , Femenino , Humanos , Factor de Crecimiento Transformador beta1/metabolismo , Virus Zika/metabolismo , Primer Trimestre del Embarazo , Trofoblastos/metabolismoRESUMEN
Species A rotaviruses (RVAs) have been recognized as one of the leading causes of acute gastroenteritis in humans worldwide. Here, the complete coding sequences of 11 RNA segments of an uncommon G9P[4] RVA strain, which was detected in feces of a diarrheal child in Japan, were determined by next-generation sequencing technology. Its genomic constellation, VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5, was determined as G9-P[4]-I2-R2-C2-M2-A2-N2-T2-E2-H2. This work reports the complete coding sequences of a G9P[4] RVA strain containing DS-1-like (genotype 2) genes that was isolated in Japan in 2013.
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Infecciones por Rotavirus , Rotavirus , Niño , Genoma Viral , Genotipo , Humanos , Japón , Filogenia , Rotavirus/genéticaRESUMEN
INTRODUCTION: Norovirus (NoV) is the most common agent causing outbreaks and sporadic cases of acute gastroenteritis among all ages, especially children under 5 years old. During the coronavirus disease 2019 (COVID-19) pandemic, NoV infection has decreased drastically in Japan due to school closures and no outbreak related to NoV infection had been reported. METHOD: In mid-September 2021, NoV outbreak occurred in kindergarten and nursery schools in Maizuru, Kyoto prefecture, Japan. Twenty-six stool samples collected from patients who were diagnosed of NoV gastroenteritis from the outbreak by an immunochromatographic (IC) kit at a pediatric outpatient clinic in Maizuru city during 3 weeks from September 13 to October 8, 2021 were examined for the presence of NoV GII by reverse transcriptase-polymerase chain reaction (RT-PCR), genome sequencing, and phylogenetic analysis. RESULT: All 26 samples were confirmed positive to NoV GII and their genotypes were identified as GII.4 Sydney[P31]. The amino acid substitutions in open reading frame1 (ORF1) and ORF2 genes were found when compared with previously detected sporadic NoV GII.4 Sydney[P31] strains isolated in Japan. The clinical characterization of infected children was described. Most of the children were mild cases and vomiting was the most frequent clinical symptom. CONCLUSION: This study reported a recent emergence of NoV GII.4 Sydney[P31] causing acute gastroenteritis outbreak in children in Japan during the COVID-19 pandemic and suggests a need for further monitoring of NoV GII.4 variants.
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COVID-19 , Infecciones por Caliciviridae , Gastroenteritis , Norovirus , COVID-19/epidemiología , Infecciones por Caliciviridae/epidemiología , Niño , Preescolar , Heces , Gastroenteritis/epidemiología , Genotipo , Humanos , Japón/epidemiología , Norovirus/genética , Pandemias , FilogeniaRESUMEN
BACKGROUND: Inadequate attention has been given to ensuring ongoing training to improve knowledge, skills and capacity of primary health care providers in low- and middle-income countries. The Hanoi Medical University, Vietnam is providing training sessions for physicians working in commune health stations (CHSs) in three mountainous, remote northern provinces in 2019. This article aims to assess these physicians' knowledge of correct medical responses to emergencies in order to assess their training needs. METHODS: We conducted a cross-sectional study amongst doctors posted to CHSs located in 3 mountainous remote provinces of northern Vietnam. We used a self-administered questionnaire that comprised questions on common medical emergencies, maternal and child care, and non-communicable disease management. We performed Chi-square tests to assess the statistical significance of differences in the mean proportions of correct answers for each health care question category, and for differences in mean proportions of correct answers by doctor characteristics. RESULTS: In total 302 doctors were recruited to the study. More than half of the sample answered 30-50% of the questions correctly, followed by around a third who answered 50-70% correctly. Less than 2% of doctors answered more than 70% correct responses to the entire question set. There were statistically significant differences between question categories, with cardiovascular care questions answered correctly significantly less often than any of the categories (p < 0.00001). CONCLUSION: The findings reported here show that the doctors who participated in the study have relatively low knowledge on common emergencies, particularly to answer cardiovascular care questions. The results also support the need for continuing medical education to improve doctors' knowledge, who are mostly practicing in resource limited remote settings.
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Educación Médica Continua , Médicos de Atención Primaria , Estudios Transversales , Urgencias Médicas , Conocimientos, Actitudes y Práctica en Salud , Humanos , Evaluación de Necesidades , Encuestas y Cuestionarios , VietnamRESUMEN
Sapovirus (SaV) is one of the pathogens related to acute gastroenteritis (AGE) in adults and children worldwide. This study reported the diversity of SaV genotypes in children with AGE in Japan from July 2014 to June 2017. Of a total of 2259 stool samples tested by using reverse transcription-PCR method and further analyzed by nucleotide sequencing, 114 (5.0%) were positive for SaV and GI.1 (83.3%) was the most predominant genotype, followed by GII.1, GIV.1, GI.2, GI.3, and GII.3 genotypes. Monthly distribution analysis demonstrated two epidemic peaks from July to December 2015 and February to May 2017. However, no detection peak was observed in 2014 and 2016. Phylogenetic analysis of the complete VP1 nucleotide sequences of these GI.1 strains revealed two major clusters of GI.1 and each of which contained GI.1 strains of both 2015 and 2017. This study suggests that the continuous surveillance of SaV is needed to monitor high genetic diversity in Japanese children with AGE.
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Infecciones por Caliciviridae/virología , Gastroenteritis/virología , Sapovirus/genética , Enfermedad Aguda , Infecciones por Caliciviridae/epidemiología , Proteínas de la Cápside/genética , Niño , Preescolar , Coinfección/epidemiología , Coinfección/virología , Heces/virología , Gastroenteritis/epidemiología , Variación Genética , Genotipo , Humanos , Japón/epidemiología , Filogenia , Prevalencia , Reinfección/epidemiología , Reinfección/virología , Sapovirus/clasificación , Estaciones del AñoRESUMEN
BACKGROUND: Acute gastroenteritis is the most common cause of illness and death in infants and young children worldwide. Rotaviruses (RVs) are the major viruses that cause acute gastroenteritis in young children, especially in developing countries in Asia and Africa. METHODS: The presence of rotavirus antigens in sera of four unvaccinated pediatric patients, aged between 4 and 6 years with severe diarrhea and dehydration, were detected by using three immunochromatographic (IC) kits. In addition, the presence of anti-rotavirus IgG, IgA, and IgM antibodies and their concentrations in patient sera were also determined by enzyme immunoassay (EIA). RESULTS: All three kits could detect rotavirus antigen in patient sera with different intensity of the test lines. When patient sera were pretreated with anti-VP6 rotavirus mouse monoclonal antibody prior to testing, the rotavirus positive test lines disappeared, suggesting that all patient sera contained VP6 protein antigen of rotavirus. Assessment of antibody concentration in these patient sera revealed that all patient sera contained IgG, IgA, and IgM antibodies against rotavirus antigen at different concentrations. CONCLUSIONS: The sensitivity of rotavirus protein detection in the patient sera of one IC kit brand was comparable to those of the EIA, suggesting this IC kit could be an alternative screening method for rapid diagnosis of rotavirus infection.
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Gastroenteritis , Infecciones por Rotavirus , Rotavirus , Animales , Anticuerpos Antivirales , Antígenos Virales , Niño , Preescolar , Heces , Gastroenteritis/diagnóstico , Humanos , Lactante , Ratones , Infecciones por Rotavirus/diagnósticoRESUMEN
INTRODUCTION: Rotavirus (RV) is the major pathogen responsible for acute gastroenteritis in infants. Since RV vaccines were introduced, a substantial decline in the incidence of severe RV infection has been reported. However, some burden still exists, even in developed countries, including Japan. METHODS: We retrospectively surveyed 380 patients hospitalized for acute gastroenteritis from 2015 to 2019. In 2019, additional detailed clinical information of 21 patients with RV gastroenteritis was obtained to evaluate the efficacy of the RV vaccine. Nine fecal samples from those patients were collected to detect the RV genotypes. RESULTS: Our data showed an increasing trend in hospitalizations for severe RV gastroenteritis in children older than 5 years. According to the Vesikari clinical severity scores in the older group (≥5 years), the gastrointestinal symptoms in vaccinated patients were less severe than those in unvaccinated patients (p = 0.014). The genotype analysis revealed that G9P[8]I1 was the major genotype in the recruited patients in 2019. CONCLUSIONS: This report warns that children older than 5 years could be affected by severe RV infection and suggests prompt intervention for this age group, similar to that in infants. In the new period in which the RV vaccine is included in Japanese national immunization programs beginning October 2020, continuous monitoring of patient clinical characteristics and RV epidemiology is required to determine the role of vaccines.
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Gastroenteritis , Infecciones por Rotavirus , Vacunas contra Rotavirus , Rotavirus , Niño , Preescolar , Heces , Gastroenteritis/epidemiología , Gastroenteritis/prevención & control , Genotipo , Hospitalización , Humanos , Lactante , Japón/epidemiología , Estudios Retrospectivos , Rotavirus/genética , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/prevención & controlRESUMEN
Multiple aspects of cornea development, including the innervation of the cornea by trigeminal axons, are sensitive to embryonic levels of thyroid hormone (TH). Although previous work showed that increased TH levels could enhance the rate of axonal extension within the cornea in a thyroxine (T4)-dependent manner, details underlying the stimulatory effect of TH on cornea innervation are unclear. Here, by examining the effects throughout all stages of cornea innervation of the two main THs, triiodothyronine (T3) and T4, we provide a more complete characterization of the stimulatory effects of TH on corneal nerves and begin to unravel the underlying molecular mechanisms. During development, trigeminal axons are initially repelled at the corneal periphery and encircle the cornea in a pericorneal nerve ring prior to advancing into the corneal stroma radially from all along the nerve ring. Overall, exogenous T3 led to pleiotropic effects throughout all stages of cornea innervation, whereas the effects of exogenous T4 was confined to timepoints following completion of the nerve ring. Specifically, exogenous T3 accelerated the formation of the pericorneal nerve ring. By utilizing in vitro neuronal explants studies we demonstrated that T3 acts as a trophic factor to directly stimulate trigeminal nerve growth. Further, exogenous T3 caused disorganized and precocious innervation of the cornea, accompanied by the downregulation of inhibitory Robo receptors that normally act to regulate the timing of nerve advancement into the Slit-expressing corneal tissues. Following nerve ring completion, the growth rate and branching behavior of nerves as they advanced into and through the cornea were found to be stimulated equally by T3 or T4. These stimulatory influences of T3/T4 over nerves likely arose as secondary consequences brought on by TH-mediated modulations to the corneal extracellular matrix. Specifically, we found that the levels of nerve-inhibitory keratan- and chondroitin-sulfate containing proteoglycans and associated sulfation enzymes were dramatically altered in the presence of exogenous T3 or T4. Altogether, these findings uncover new roles for TH on corneal development and shed insight into the mechanistic basis of both T3 and T4 on cornea innervation.
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Axones/efectos de los fármacos , Córnea/inervación , Desarrollo Embrionario/fisiología , Tiroxina/farmacología , Triyodotironina/farmacología , Animales , Embrión de Pollo , Córnea/efectos de los fármacos , Córnea/embriología , FemeninoRESUMEN
Solid-state nanopores constitute a versatile platform for study of ion transport in nanoconfinement. The electrical double layer (EDL) plays a vital role in such nanoconfinements, but effects of induced surface charge on the EDL in the presence of an external transmembrane electric field are yet to be characterized. Here, the formation of induced charge on the nanopore sidewall surface and its effects, via modulation of the EDL and electroosmotic flow, on the ionic current are elucidated using a novel experimental setup with solid-state truncated-pyramidal nanopores. This study consists of three complementary approaches, i.e., an analytical model for induced surface charge, numerical simulation of induced surface charge, electroosmotic flow, and ionic current, and experimental validation with respect to the ionic current. The induced surface charge is generated by polarization in the dielectric membrane as a response to the applied electric field. This charge generation results in a nonuniform density of surface charge along the nanopore sidewall. It further causes ions in the electrolyte to redistribute, leading to a massive accumulation of single-polarity ions in the EDL and their counterions near the smaller opening of the nanopore. It also alters electrohydrodynamic properties in the nanopore, giving rise to the formation of electroosmotic vortexes in the vicinity of the smaller opening of the nanopore. Finally, the pattern of the electroosmotic flow can significantly influence the transport properties of the nanopore.
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OBJECTIVES: Value and health technology assessment (V/HTA) is often used in clinical, access, and reimbursement decisions. V/HTA data-source selection may not be transparent, which is a necessary element for stakeholder understanding and trust and for fostering accountability among decision makers. Peer review is considered one mechanism for judging data trustworthiness. Our objective was (1) to use publicly available documentation of V/HTA methods to identify requirements for inclusion of peer-reviewed evidence sources, (2) to compare and contrast US and non-US approaches, and (3) to assess evidence sources used in published V/HTA reports. METHODS: Publicly available methods documentation from 11 V/HTA organizations in North America and Europe were manually searched and abstracted for descriptions of requirements and recommendations regarding search strategy and evidence-source selection. The bibliographies of a subset of V/HTA reports published in 2018 were manually abstracted for evidence-source types used in each. RESULTS: Heterogeneity in evidence-source retrieval and selection was observed across all V/HTA organizations, with more pronounced differences between US and non-US organizations. Not all documentation of organizations' methods address the evidence-source selection processes (7 of 11), and few explicitly reference peer-reviewed sources (3 of 11). Documentation of the evidence-source selection strategy was inconsistent across reports (6 of 13), and the level of detail provided varied across organizations. Some information on evidence-source selection was often included in confidential documentation and was not publicly available. CONCLUSIONS: Disparities exist among V/HTA organizations in requirements and guidance regarding evidence-source selection. Standardization of evidence-source selection strategies and documentation could help improve V/HTA transparency and has implications for decision making based on report findings.
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Documentación/normas , Revisión por Pares , Evaluación de la Tecnología Biomédica/métodos , Europa (Continente) , Humanos , América del NorteAsunto(s)
Gastroenteritis , Rotavirus , Niño , Humanos , Lactante , Gastroenteritis/diagnóstico , Pruebas Inmunológicas , HecesRESUMEN
Multiplex RT-PCR method using five sets of panel primers was developed for the detection of diarrheal viruses, including rotavirus A, B, and C, adenovirus, astrovirus, norovirus GI and GII, sapovirus, Aichi virus, parechovirus, enterovirus, cosavirus, bocavirus, and Saffold virus. The sensitivity of the method was evaluated and tested with 751 fecal specimens collected from Japanese children with acute diarrhea. Several kinds of viruses were detected in 528 out of 751 (70.3%) fecal specimens. Mixed-infection with different viruses in clinical specimens could also be effectively detected. The method proved to be reliable with highly sensitive and specific and useful for routine diagnosis. J. Med. Virol. 89:818-824, 2017. © 2016 Wiley Periodicals, Inc.
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Diarrea/virología , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Virosis/diagnóstico , Virus/clasificación , Virus/aislamiento & purificación , Adolescente , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Japón , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad , Factores de Tiempo , Virus/genéticaRESUMEN
Enterovirus D68 (EV-D68) is known to be causative agent of mild to severe upper and lower respiratory illnesses in sporadic cases and outbreaks. We present a case report of a 3-month-old child with acute gastroenteritis who visited a pediatric clinic in Kyushu area in Japan in 2015. A stool sample collected from the patient was screened for diarrheal viruses by multiplex RT-PCR. The result showed that the sample was positive only for enterovirus, and EV-D68 clade B3 was identified by sequence analysis of the viral protein 1 gene. This study supports an association between EV-D68 infection and acute gastroenteritis.
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Enterovirus Humano D/aislamiento & purificación , Gastroenteritis/microbiología , Enterovirus , Humanos , Lactante , JapónRESUMEN
BACKGROUND: Acute encephalitis is a serious neurological condition having a high mortality rate and affecting both children and adults. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the major viral causes of acute encephalitis/ encephalopathy in Asia. METHODS: Using previously published primers that have been widely used to screen for herpes virus-6, influenza A virus, human parechovirus, herpes simplex viruses 1 and 2, Japanese encephalitis virus, group A rotavirus, enterovirus, adenovirus, and dengue virus in clinical samples, a single-tube multiplex PCR assay was developed and was tested for its sensitivity and specificity. The method was then applied to screen 57 clinical samples, consisting of 13 fecal samples, 5 throat swabs, 3 post-nasal swabs, 18 serum samples, and 18 cerebrospinal fluid (CSF) samples, collected from 18 hospitalized Japanese children with suspected viral encephalitis/encephalopathy for the target viruses, and the results were compared with those of a monoplex PCR method. RESULTS: Positive viral controls of the 10 viruses were correctly typed using this multiplex PCR method. The multiplex PCR method showed high specificity with no unspecific amplification to non-target viruses. The results of applying this PCR method for screening clinical samples showed that 6 fecal samples, 2 serum samples, and 1 CSF sample collected from 7 patients were positive for a virus, specifically group A rotavirus (4 patients, 22.2%), enterovirus (2 patients, 11.1%), or adenovirus (1 patient, 5.6%). In comparison with monoplex PCR, for group A rotavirus, enterovirus, and adenovirus, the sensitivity of this multiplex PCR method decreased for serum, cerebrospinal fluid, and throat swab samples. CONCLUSIONS: This newly developed multiplex PCR method is a simple, rapid diagnostic tool and can be used to screen clinical samples for viruses causing acute encephalitis/encephalopathy in children in Asian countries.
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ADN Viral/genética , Encefalitis Viral/diagnóstico , Reacción en Cadena de la Polimerasa Multiplex , Virus/genética , Enfermedad Aguda , Calibración , Niño , Cartilla de ADN , Encefalitis Viral/virología , Humanos , Japón , Reacción en Cadena de la Polimerasa Multiplex/normas , Valor Predictivo de las Pruebas , Estándares de Referencia , Reproducibilidad de los Resultados , Virus/clasificaciónRESUMEN
BACKGROUND: High-mobility group box 1 (HMGB1), a DNA-binding protein, has recently been shown to have effects on HIV replication, but the effects are dependent on the cell type and the timing of infection. Using human primary T cells, this study aimed to investigate the role of HMGB1 in HIV-1 replication in newly infected cells. METHODS: Human primary T cells were infected with the HIV-1 LAI (X4) strain and then cultured in the presence of recombinant HMGB1 protein or an anti-HMGB1 antibody at various concentrations. At the indicated time points, HIV-1 p24 concentrations in the culture media were measured by ELISA. Cell proliferation, basal HMGB1 concentration, and CD3, CD4, CXCR4, and receptor for advanced glycation end products (RAGE) expression were also examined. RESULTS: Recombinant HMGB1 could enhance HIV replication in newly infected primary T cells. In the presence of an anti-HMGB1 antibody (5 µg/mL or higher), significantly lower concentrations of HIV-1 p24 were observed in the cultures of primary T cells during the post-infection period. CONCLUSIONS: The data presented suggest that HMGB1 plays a role in the enhancement of HIV-1 replication in newly infected T cells. This finding provides useful information toward understanding HIV pathogenesis and for the development of new therapeutic strategies.
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Infecciones por VIH/virología , VIH-1/crecimiento & desarrollo , Proteína HMGB1/metabolismo , Linfocitos T/virología , Replicación Viral , Anticuerpos/farmacología , Proliferación Celular , Células Cultivadas , Proteína p24 del Núcleo del VIH/metabolismo , Infecciones por VIH/genética , Infecciones por VIH/metabolismo , VIH-1/efectos de los fármacos , VIH-1/metabolismo , Proteína HMGB1/antagonistas & inhibidores , Interacciones Huésped-Patógeno , Humanos , Cultivo Primario de Células , Transducción de Señal , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismo , Factores de Tiempo , TransfecciónRESUMEN
BACKGROUND: Infection with Campylobacter jejuni and C. coli are recognized as the major cause of bacterial gastroenteritis in humans. METHODS: A total of 310 fecal samples collected from Thai adult patients with diarrhea in 2008 were screened for the presence of Campylobacter by PCR. Resistance to fluoroquinolone and macrolides of the detected Campylobacter strains were analyzed by studying the mutations in the gyrA and 23S rRNA genes, respectively. RESULTS: Campylobacter species were detected in 4/310 (1.3%) of diarrheal patients, and C. jejuni was found in 3 of the 4 cases (75%). Fluoroquinolone resistance was noted in 2 cases (50%); however, no resistance to macrolides was observed. CONCLUSIONS: Campylobacter was detected in a low prevalence in adult Thai patients hospitalized with diarrhea, and the resistance to fluoroquinolones is still a matter of concern in case antibiotic therapy is required.