Hiroko Sakuma

Prognostic predictive values of serum cytochrome c , cytokines, and other laboratory measurements in acute encephalopathy with multiple organ failure

Aims: To evaluate the prognostic predictive values of cytochrome c, cytokines, and other laboratory measurements in serum collected during neurological onset in acute encephalopathy with multiple organ failure. Methods: In addition to general laboratory examinations, the concentrations of cytochrome c (apoptosis marker) and cytokines (inflammatory markers) were measured in serum samples collected at the initial phase in 29 patients with acute encephalopathy. The obtained values were evaluated as predictors for the development of severe encephalopathy. Results: Cytochrome c, tumour necrosis factor α (TNF-α), interleukin 6 (IL-6), soluble TNF-receptor 1 (sTNF-R1), and aspartate aminotransferase (AST) concentrations at the initial phase were high and correlated well with patient outcome. High concentrations of serum cytochrome c (>45 ng/ml), sTNF-R1 (>2000 pg/ml), AST (>58 IU/dl), IL-6 (>60 pg/ml), and TNF-α (>15 pg/ml) predicted an unfavourable prognosis (sequelae and death) at 93%, 79%, 82%, 77%, and 60%, respectively. The specificity of those markers was 100%, 89%, 83%, 100%, and 100%, respectively. Conclusions: Serum cytochrome c is the most sensitive and specific predictor for the development of severe encephalopathy at the initial phase. Results suggest that this marker might be used to guide decisions regarding the start of the initial treatment and further intensive care.

Human influenza virus infection and apoptosis induction in human vascular endothelial cells.

Acute encephalopathy accompanying influenza virus infection results in brain and systemic organ failure mainly through vasogenic edema with high levels of inflammatory cytokines, such as blood tumor necrosis factor (TNF)‐α and interleukin (IL)‐6, as well as the cytochrome c apoptosis marker. A highly virulent strain of avian influenza virus causes fatal infection in chickens by infecting vascular endothelial cells in systemic organs, inducing apoptosis therein. To verify the possibility of apoptosis induction by human influenza virus in infected human vascular endothelial cells, purified influenza virus‐infected human umbilical vein endothelial cells (HUVECs) were examined using a tissue culture method. When pre‐treated with TNF‐α, influenza virus (Philippine strain, H3N2) promoted TNF‐α induced apoptosis of HUVECs. Viral replication was confirmed in HUVECs infected with the Philippine strain in the absence of TNF‐α by measurement of the amount of infective virus in the culture supernatant using the tissue culture infectious dose (TCID) method, immunohistochemistry and real‐time PCR. The number of influenza virus genomes in the infected HUVECs at 24 hr post‐infection increased about fivefold compared to that just after virus adsorption. Many TUNEL‐positive influenza virus‐infected HUVECs were observed using the TUNEL method. Furthermore, cleaved caspase 3 was also detected in influenza virus‐infected cells by immunofluorescence staining. These results demonstrated that human influenza virus can infect and replicate in human vascular endothelial cells and induce apoptosis therein. J. Med. Virol. 80:1072–1078, 2008.

Application of polymerase chain reaction and subsequent phylogenetic analysis to the diagnosis of enteroviral infection in the central nervous system

BACKGROUND Enteroviral infections of the central nervous system (CNS) are often difficult to diagnose, even if consistent conventional laboratory methodologies are used. OBJECTIVES To clarify the efficiency of two polymerase chain reaction (PCR) methods for the sensitive detection of enteroviruses and for the identification of enteroviral genotypes based on phylogenetic analysis of the amplified genome sequences, and to facilitate the diagnosis of enteroviral infection in CNS. STUDY DESIGN Cerebrospinal fluid (CSF), throat swab, rectal swab, and/or serum samples were collected from 171 patients with aseptic meningitis and 67 patients with febrile seizures. The samples were tested for the presence of enteroviruses by cell culture and PCR methods for the detection and identification of enteroviruses. RESULTS In 111 (64.9%) of 171 patients with aseptic meningitis, enteroviruses were isolated by cell cultures from any site. In 143 (83.6%) patients, including 110 of 111 patients with aseptic meningitis, the enteroviral genome was detected in CSF by PCR. No enterovirus was isolated from any site for the 67 patients with febrile seizures. PCR detected the enteroviral genome in CSF samples from 13 (61.9%) of 21 patients who developed febrile seizures in the summer (June-August). Phylogenetic analysis of amplified genome sequences showed that the major pathogens of febrile seizures in summer were group A coxsackieviruses, which are usually difficult to isolate by cell culture. CONCLUSION PCR methods for the detection and identification of enteroviruses were useful for the diagnosis of enteroviral infection in CNS.

Functional polymorphism of the promoter region of the prostacyclin synthase gene and severity of RSV infection in hospitalized children

Prostaglandin I2 (PGI2) protects against RSV‐induced illness in mice. A variable‐number tandem repeat (VNTR) polymorphism has been detected in the promoter region of the PGI2 synthase (PGIS) gene. We sought to determine if PGI2 concentrations or polymorphisms of the PGIS gene correlate with severity of RSV lower respiratory tract infections (LRTI) in human infants. VNTR polymorphisms were studied in 81 previously healthy children between birth and 12 months of age who were hospitalized for LRTI due to RSV and 98 healthy adult control subjects. The severity of RSV infection was quantified using a clinical scoring system, and infant urine samples were collected during the acute illness for measurement of the urinary metabolite of PGI2. There were no significant differences in the overall distribution of alleles and genotypes between infants with RSV LRTI and the control subjects. The severity of RSV infection significantly inversely correlated with urinary PGI2 metabolite concentrations. The urinary PGI2 metabolite concentration correlated with the number of VNTR. The presence of a genotype with a low number VNTR repeats significantly correlated with the most severe RSV LRTI, and genotypes with the highest number of VNTR correlated with the least severe RSV LRTI. A functional polymorphism in the promoter region of the PGIS gene is associated with both significant differences in urinary PGI2 concentrations during RSV LRTI, and severity of RSV infection in previously healthy infants. J. Med. Virol. 80:2015–2022, 2008.

Influenza Viral Load and Peramivir Kinetics after Single Administration and Proposal of Regimens for Peramivir Administration against Resistant Variants

ABSTRACT We estimated the efficacy of the current single administration of peramivir on the basis of peramivir pharmacokinetics in the upper respiratory tract (URT) and determined the predictive peramivir concentration-time curve to assess its efficacy against viruses with decreased susceptibility to neuraminidase inhibitors. Serum, nasal swab, or aspiration samples were collected from 28 patients treated with 10 mg/kg body weight peramivir. The sequential influenza viral RNA load and susceptibility after peramivir administration were measured using a quantitative real-time reverse transcription-PCR and neuraminidase inhibition assay. The peramivir concentrations in the serum and URT after a single administration at 10 mg/kg were measured, and the predictive blood and URT peramivir concentration-time curves were determined to assess various administration regimens against resistant variants. The peramivir concentration decreased to <0.1% of the maximum concentration of drug in serum (Cmax) at 24 h after administration. Rapid elimination of peramivir from the URT by 48 h after administration may contribute to an increase in the influenza A viral load after day 3 but not to a decrease in the influenza B viral load, despite the absence of a decrease in the susceptibility to peramivir. A longer maintenance of a high level of peramivir in the URT is expected by divided administration rather than once-daily administration. When no clinical improvement is observed in patients with normal susceptibility influenza A and B, peramivir readministration should be considered. In severe cases caused by resistant variants, better inhibitory effectiveness and less frequent adverse events are expected by divided administration rather than once-daily administration with an increased dosage.

Incidence and index of severity of hemolytic uremic syndrome in a 26 year period in Fukushima Prefecture, Japan

There have been a number of reports on large outbreaks of hemolytic uremic syndrome (HUS), but there have been no long‐term studies of sporadic HUS in Japan. This study therefore investigated the epidemiology and prognosis of HUS in Fukushima Prefecture over a 26 year period.

Uteroglobulin-related protein 1 and severity of respiratory syncytial virus infection in children admitted to hospital.

There are several reports suggesting that genetic factors contribute to the severity of infection with the respiratory syncytial virus (RSV). Infants hospitalized with lower respiratory tract infection (LRTI) due to RSV are at a significantly increased risk for both recurrent wheezing and childhood asthma. Uteroglobin‐related protein 1 (UGRP1) is a secretory protein expressed in the airways, and speculated to have anti‐inflammatory activity. The presence of the −112G/A polymorphism in the UGRP1 promoter was found to have a significant correlation with asthma phenotype. Also plasma UGRP1 levels were shown to be associated both with this polymorphism and the severity of asthma. The study population consisted of 62 previously healthy infants, ≤12 months of age, who were hospitalized with RSV LRTI, and a control group of 99 healthy adults. Genotyping was performed by restriction fragment length polymorphism. UGRP1 serum levels were determined using ELISA. There were no significant differences in the overall distribution of UGRP1 −112G/A polymorphism genotypes or alleles between the hospitalized infants and healthy adults. A comparison of serum UGRP1 concentration measured at the time of admission and discharge between patients with and without the −112A allele revealed that there was no relation between the presence of the −112A allele and serum UGRP1 in hospitalized infants with RSV infection. Furthermore, there was no relationship between severity of RSV infection and genotype or serum UGRP1 concentration. These results suggest that UGRP1 does not have a major role in the development of severe RSV infection. J. Med. Virol. 83:1086–1092, 2011.