Clinical and Virologic Impacts of Respiratory Viral Co-infections in Children With Influenza.

BACKGROUND: Advances in multiplex polymerase chain reaction (PCR) methods have enabled the simultaneous detection of multiple respiratory viruses. We aimed to estimate the clinical and virologic impacts of influenza and other respiratory virus co-infection in children. METHODS: We enrolled 38 and 35 children diagnosed with influenza and treated with baloxavir marboxil (baloxavir) and oseltamivir, respectively. We performed quantitative reverse transcription-PCR to detect and measure the levels of noninfluenza viruses from 3 nasopharyngeal swab samples collected before and on days 3 and 5 after the initial antiviral dose. We assessed patients' clinical information using questionnaires. RESULTS: One or more respiratory viruses other than influenza virus were detected in 26 (35.6%) of 73 children before antiviral treatment. The influenza virus load and clinical characteristics on the day of influenza onset were similar between children with and without virus co-infections. Of the 26 and 32 children without the emergence of the reduced baloxavir and oseltamivir susceptible variants after treatment, 8 (30.8%) and 7 (21.9%) children were dually co-infected with human rhinovirus only, respectively. The level of human rhinovirus RNA on day 0 in these children was less than -3 log 10 that of influenza virus RNA, and the human rhinovirus co-infection had no impact on the disease course either clinically or virologically. CONCLUSIONS: When multiple respiratory viruses are detected in the same patient, it is necessary to assess clinical symptoms as well as the levels of detected viruses to determine which virus contributes to the development of illness.

Detection of Variants With Reduced Baloxavir Marboxil and Oseltamivir Susceptibility in Children With Influenza A During the 2019-2020 Influenza Season.

BACKGROUND: We aimed to detect influenza variants with reduced susceptibility to baloxavir marboxil (baloxavir) and oseltamivir and identify differences in the clinical course between children with and without these variants after antiviral treatment. METHODS: During the 2019-2020 influenza season, we enrolled children with confirmed influenza A (20 treated with baloxavir and 16 with oseltamivir). We analyzed patients' sequential viral RNA loads and infectious virus titers, the drug susceptibilities of clinical isolates, and amino acid substitutions in the viral polymerase acidic protein subunits or neuraminidase. We assessed patients' clinical information using questionnaires. RESULTS: All viral RNA loads and virus titers were significantly decreased after treatment, but we detected baloxavir-resistant and oseltamivir-resistant variants in 5 of 20 and 3 of 16 patients, respectively. The duration of fever was similar between patients with and without the variants, but infectious viral shedding lasted 3 days longer in patients with baloxavir-resistant variants. In addition, the duration to improvement of clinical symptoms was longer in these patients (75.0 vs 29.5 hours; P = .106). CONCLUSIONS: After antiviral treatment, the emergence of baloxavir-resistant variants may affect the patients' clinical course, but oseltamivir-resistant variants had no clinical impact.

Detection of Variants With Reduced Baloxavir Marboxil Susceptibility After Treatment of Children With Influenza A During the 2018-2019 Influenza Season.

During the 2018-2019 influenza seasons, we detected reduced baloxavir marboxil (baloxavir) susceptible variants with I38S or I38T amino acid substitutions on the PA subunit of influenza virus ribonucleic acid polymerase in 7 of 18 baloxavi-treated children and found that virus titer rebounded in some of these children with variants. We also found fever durations to be similar between patients with or without the variants, but the patients with variants shed the virus 3 days longer and took longer to improve clinical symptoms than those without variants. The emergence of these variants should be monitored during future influenza seasons.

Successful therapy with tonsillectomy plus pulse therapy for the relapse of pediatric IgA nephropathy treated with multi-drugs combination therapy.

Immunoglobulin A nephropathy (IgAN) is the most common form of chronic glomerulonephritis worldwide.　In Japan, the treatment for use as an initial therapy was established in Guidelines for the Treatment of Childhood IgA nephropathy; however, no rescue therapy for recurrent or steroid-resistant pediatric IgAN was established.　We report here a 15-year-old boy with severe IgAN, who was treated with combination therapy involving prednisolone, mizoribine, warfarin, and dilazep dihydrochloride for 2 years.　The response to the combination therapy was good and both proteinuria and hematuria disappeared.　The pathological findings at the second renal biopsy were improved and PSL was discontinued.　However, due to nonadherence to the treatment regimen and tonsillitis, macrohematuria and an increase of proteinuria were again observed and the pathological findings at the third renal biopsy showed clear deterioration.　The patient was, therefore, diagnosed with recurrent IgAN.　Tonsillectomy plus methylprednisolone pulse therapy (TMP) was performed as a rescue therapy for the recurrence of severe IgAN.　Both the proteinuria or hematuria subsequently disappeared, and no proteinuria or hematuria has been observed and kidney function has remained normal during a 5-year follow-up.　The patient experienced no severe side effects associated with the drug regimens.　In conclusion, our case suggests that TMP may be an effective and useful rescue therapy for recurrent IgAN after multi-drug combination therapy.

A case of a child with sle presenting with hps as a primary manifestation.

The primary manifestations of systemic lupus erythematosus (SLE) are various. One such manifestation is hemophagocytic syndrome (HPS). We here report a child with SLE presenting with HPS as a primary manifestation. In October 2010, an 11-year-old Japanese boy presented with pancytopenia, elevated liver enzymes, hyperferritinemia and hemophagocytosis due to macrophages in the bone marrow, and was diagnosed with HPS. A year later, he was found to have proteinuria and hematuria. Oral aphtha and Raynaud's phenomenon were observed, and the patient showed low serum complement levels and was positive for anti-nuclear antibodies (ANAs). He was subsequently diagnosed with SLE. Moreover, low serum complement levels and ANA positivity were detected in a serum sample preserved at the onset of HPS. The HPS was considered to be a primary manifestation of SLE on the basis of these findings. Based on this case, the presence of an underlying disease, such as SLE, should be investigated in cases of HPS.

Efficacy of recombinant human soluble thrombomodulin for childhood hemolytic uremic syndrome.

Recombinant human soluble thrombomodulin (rhTM) is a promising therapeutic natural anticoagulant and is used clinically for the treatment of disseminated intravascular coagulation (DIC). Herein is reported the cases of two HUS children treated with rhTM. The patients were diagnosed as having typical HUS on the basis of thrombocytopenia, hemolytic anemia, acute renal failure, and the detection Escherichia coli 0157. I.v. rhTM was started as an anti-coagulant drug. At 2 days after the first treatment in both patients, fibrin/fibrinogen degradation products and d-dimer levels were significantly decreased, and there was a subsequent slight improvement in thrombocytopenia, and a decrease in serum lactate dehydrogenase level. Urinary protein excretion gradually diminished and a decrease in serum creatinine level was observed. The patients did not require dialysis therapy. The present results suggest that rhTM may be a safe and effective treatment for DIC complicated with HUS in children.

Relapse of minimal change nephrotic syndrome after intravitreal bevacizumab.

Reported herein is a case of relapse of nephrotic syndrome (NS) after intravitreal injection of bevacizumab, a monoclonal antibody that binds to vascular endothelial growth factor (VEGF), in a 16-year-old girl. She had a diagnosis of steroid-dependent NS and had been treated with prednisolone, and remained in remission. The patient had had visus brevior 10 years previously, and was diagnosed with severe myopic choroidal neovascularization (mCNV). Intravitreal bevacizumab was given for mCNV. At 9 days after intravitreal injection of bevacizumab, proteinuria was positive. The patient had relapse of NS caused by bevacizumab, and steroid pulse therapy was then given and the proteinuria resolved. It is necessary to take particular care to prevent NS relapses in patients with mCNV treated with intravitreal bevacizumab.

Treatment with oseltamivir and prone positioning for a 9-year-old boy with ARDS caused by pandemic H1N1 2009 influenza.

Here we report the case of a 9-year-old boy with acute respiratory distress syndrome (ARDS) caused by novel H1N1 swine-origin influenza virus A. A diagnosis of ARDS caused by a novel influenza A (H1N1) virus was made on the basis of chest X-ray and computed tomography together with low oxygenation index (OI) and the detection of novel influenza A (H1N1) virus from tracheal secretion samples. Oseltamivir phosphate and prone positioning were effective in the treatment of ARDS in this case. These findings suggest that anti-viral drugs and prone positioning can play an important role in the improvement of ARDS caused by novel H1N1 swine-origin influenza virus A.

Development of common variable immunodeficiency in IgA- and IgG2-deficient patients with systemic lupus erythematosus.

BACKGROUND: There have been few reports on children who developed common variable immunodeficiency (CVID) in association with immunoglobulin A (IgA) and IgG2 deficiencies and systemic lupus erythematosus (SLE). CASE-DIAGNOSIS/TREATMENT: Our patient experienced nephrotic syndrome and acute respiratory distress syndrome (ARDS) caused by influenza A/H1N1 virus infection at 5 years of age. A diagnosis of IgA and IgG2 deficiency and SLE was made on the basis of severe proteinuria, hematuria, hypocomplementemia, high anti-DNA antibody and antinuclear antibody (ANA) titers, and malar rash. However, these clinical signs and symptoms and laboratory features disappeared after the administration of methylprednisolone pulse therapy and prednisolone. For the 5 years following the initial treatment for SLE, the patient experienced a number of infections and had a low serum total IgG level; she was eventually diagnosed with CVID. The administration of intravenous immunoglobulin (IVIG) was required to prevent subsequent infections, and no relapse of SLE was observed. CONCLUSION: We report the development of CVID in an IgA- and IgG2-deficient patient with SLE on the basis of multiple episodes of infection. To prevent the development of CVID in IgA- and IgG2-deficient patients with SLE, it is important to prevent immune dysregulation by the avoidance of infections through the use of IVIG therapy.

A case of Henoch-Schoenlein purpura nephritis recurring after 3 years remission.

Henoch-Schoenlein purpura (HSP) is a systemic disorder characterized by leukocytoclastic vasculitis involving the capillaries with IgA immune complexes deposition, and about 7% of patients with HSP experience recurrence. Most patients with recurring of HSP nephritis show a recurrence of clinical symptoms over a period ranging from 2 to 5 months, even after the disappearance of initial symptoms. Here we report a 9-year-old girl diagnosed with recurrent HSP and severe crescentic glomerulonephritis 3 years after complete resolution of the initial symptoms of HSP. Our case is unique in respect of the recurrence at more than 3 years after the complete resolution of initial symptoms, suggesting that careful followup is required in spite of improved renal symptoms in cases of HSP.

Tonsillectomy with methylprednisolone pulse therapy as rescue treatment for steroid-resistant IgA nephropathy in children.

Primary immunoglobulin A (IgA) nephropathy is characterized by microhematuria and proteinuria and by the deposition of IgA in the glomerular mesangium. Steroid was a main drug for treatment of IgA nephropathy. However, some of children with IgA nephropathy are resistance to steroid treatment, but the therapy for steroid-resistant IgA nephropathy was not established. There have been reports on the efficacy of tonsillectomy as an initial treatment for IgA nephropathy in adults and children. We examined whether tonsillectomy with methylprednisolone pulse therapy (tonsillectomy pulse therapy) was effective as rescue treatment for steroid-resistant pediatric IgA nephropathy. We studied 11 patients (age at onset and duration of follow-up, 11.7 +/- 2.0 and 6.2 +/- 1.1 years) who had been diagnosed with steroid-resistant IgA nephropathy. Clinical features, laboratory data, and pathological findings were retrospectively compared between before and after tonsillectomy pulse therapy. Urinary protein excretion was significantly decreased at 24.7 +/- 7.3 months after tonsillectomy pulse therapy. On renal pathologic examination of 6 patients who underwent renal biopsy at 17.1 +/- 6.9 months after tonsillectomy pulse therapy, the activity index, an index of inflammation, was lower compared to the index evaluated before the therapy, but the chronic index, an index of renal sclerosis, remained unchanged. At 24.7 +/- 7.3 months after tonsillectomy pulse therapy, seven patients had normal urine and four had minor urinary abnormalities; namely, none had active renal disease or renal insufficiency. Our findings suggest that tonsillectomy pulse therapy may be effective as rescue treatment for steroid-resistant IgA nephropathy in childhood.

Accumulation of macrophages expressing myeloid-related protein 8 associated with the progression of sclerotic changes in children with IgA nephropathy.

Myeloid-related protein (MRP) 8 is a calcium-binding protein of the S100 family. The renal accumulation of macrophages expressing MRP8 is associated with the inflammatory activity of glomerulonephritis. We evaluated the renal accumulation of macrophages expressing MRP8 in children with IgA nephropathy (IgAN). We collected data on 25 IgAN children who had been treated with prednisolone and divided these patients into two groups: Favorable group, consisting of 11 patients with normal urine and 6 with minor urinary abnormalities at 4.3 +/- 1.3 years after initial treatment; and Unfavorable group, consisting of 8 patients with persistent nephropathy. The pathological renal findings were compared between both groups. The second biopsy was performed at two years after first biopsy at 5.5 +/- 4.9 months from onset. In Favorable group, the glomerular accumulation of macrophages expressing MRP8, and mesangial cells expressing alpha-smooth muscle actin (alpha-SMA) were lower in the second biopsy specimens than those of the first biopsy specimens. In Unfavorable group, the glomerular accumulation of macrophages expressing MRP8 detected in the second biopsy specimens was similar to that of the first biopsy, while the number of mesangial cells expressing alpha-SMA and the index of renal sclerosis were higher in the second biopsy than in the first biopsy. The indexes of renal sclerosis were higher in children with more macrophages expressing MRP8 than in children with less macrophages expressing MRP8. Our results suggest that renal macrophages expressing MRP8 may be involved in the progression of sclerotic changes in children with IgAN.

Uninephrectomy induces progressive glomerulosclerosis and apoptosis in anti-Thy1 glomerulonephritis.

Administration of the anti-Thy1 antibody in rats induces reversible glomerulonephritis resembling human mesangiolytic and mesangioproliferative diseases. The purpose of the present study was to design a model of irreversible glomerulosclerosis, using the anti-Thy1 antibody injection after uninephrectomy, and examine it, focusing on apoptosis in the process of progressive sclerotic changes. Wistar rats were divided into three groups: one-kidney groups (group I and III) and a two-kidney group (group II). All groups were injected with the anti-Thy1 antibody (OX-7) at day 0, and group I and III were uninephrectomized at day -6. Only group III rats were given a half dose of OX-7 as compared with group I and II. Rats were killed for histological examinations at days 7, 14 and 30. In group I, progressive glomerular lesions, such as glomerular adhesion to Bowman's capsule, crescent formation, and collapse of capillary tufts were observed at days 14 and 30. No significant differences were observed in the pathological findings between group I and III. There was a significantly higher number of glomerular terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling-positive cells in group I as compared to group II at days 7 and 14. Moreover, the glomerular expression of transforming growth factor-beta, heparan sulfate proteoglycan and chondroitin sulfate proteoglycan significantly increased in group I as compared to group II at days 7 and 14. Progressive glomerulosclerosis can be induced in the rat by a single injection of the anti-Thy1 antibody after unilateral nephrectomy. It is suggested that apoptosis and extracellular matrix accumulation play an important role in the development of glomerulosclerosis.

FB21, a monoclonal antibody that reacts with a sialic-acid-dependent carbohydrate epitope, is a marker for glomerular endothelial cell injury.

BACKGROUND: FB21 is reactive with glomerular endothelial cells and distal tubules of the human kidney and is bound to a sialic-acid-dependent cell-surface antigen. We evaluated FB21 staining in fetal kidneys and kidneys of children and adults with normal kidneys and glomerulonephritis and investigated whether FB21 can be used as a marker for endothelial cell injury. METHODS: This study was performed on 6 children, 10 adults, and 12 fetuses with normal kidneys and 113 patients diagnosed with primary and secondary glomerulonephritis. We evaluated renal staining for FB21 in children with normal kidneys and glomerulonephritis and measured serum E-selectin concentrations in patients with hemolytic uremic syndrome (HUS) and Henoch-Schönlein purpura nephritis (HSPN). RESULTS: (1) FB21 was reactive with endothelial cells of normal kidneys and detected on the surface of endothelial cells by immunoelectron microscopy. (2) FB21 was reactive with endothelial cells in kidneys of fetuses older than 32 weeks. (3) Endothelial cell FB21 staining scores in the first renal biopsy specimens of patients with HUS and HSPN were lower than those in normal kidneys of children and correlated negatively with serum E-selectin concentrations. (4) Endothelial cell FB21 staining of crescentic and sclerotic glomerular lesions in patients with immunoglobulin A nephropathy, membranoproliferative glomerulonephritis, and focal glomerulosclerosis was weaker than that in normal kidneys. CONCLUSION: These results suggest that FB21 can be used as a marker for glomerular endothelial cell injury in various types of glomerulonephritis.

Evaluation of T helper-1/-2 balance on the basis of IgG subclasses and serum cytokines in children with glomerulonephritis.

BACKGROUND: To clarify the mechanism of deposition of immunoglobulin G (IgG) subclasses in glomerulonephritis in children, we investigated IgG subclasses in glomerular deposits and T helper subtype 1 (T(H)1)/T(H)2 cytokine balance in pediatric patients with glomerulonephritis. METHODS: We enrolled 95 children in whom glomerulonephritis had been diagnosed in our hospital between 1993 and 2000. Patients were divided into 5 groups according to histological diagnosis: 31 patients with lupus nephritis (LN), 22 patients with membranoproliferative glomerulonephritis (MPGN), 7 patients with membranous glomerulonephritis (MGN), 20 patients with Henoch-Schönlein purpura nephritis, and 20 patients with IgA nephropathy. We compared serum IgG subclass values, serum cytokine (interleukin-2 [IL-2] receptor [IL-2R], IL-2, IL-4) values, and immunofluorescence evidence of glomerular IgG subclasses in the kidney among groups. RESULTS: (1) High serum IgG1 and IgG2 values and glomerular IgG1 and IgG2 deposits were found frequently in the LN group. (2) High serum IgG3 values and glomerular IgG3 deposits were found frequently in the MPGN group. (3) High serum IgG4 values and glomerular IgG4 deposits were found frequently in the MGN group. (4) Conversely, cytokine measurements showed high serum IL-2 and IL-2R values in the LN and MPGN groups, and serum IL-4 values were high in the MGN group. CONCLUSION: These findings suggest that the pathogenetic mechanism of LN may involve both the T(H)1 and T(H)2 pattern, the pathogenetic mechanism of MPGN may involve the T(H)1 pattern, and the pathogenetic mechanism of MGN may involve the T(H)2 pattern.

Efficacy of prednisolone and mizoribine therapy for diffuse IgA nephropathy.

OBJECTIVE: There have been only a few studies concerning oral prednisolone and mizoribine therapy for diffuse IgA nephritis (IgAN). We evaluated the efficacy of prednisolone and mizoribine therapy for diffuse IgAN. METHODS: We enrolled 34 patients who had been diagnosed as having diffuse IgAN with severe proteinuria during the period from 1992 to 1999. Following diagnostic renal biopsy, the patients were treated with prednisolone, mizoribine, warfarin and dilazep dihydrochloride. The clinical features, laboratory data and pathological findings between pre- and post-therapy were investigated. RESULTS: The mean urinary protein excretion after 6 months of treatment had decreased significantly compared to pre-therapy. The incidence of hematuria in post-therapy was lower than that of pre-therapy. The grading index decreased significantly from 4.8 +/- 2.1 at the first biopsy to 2.3 +/- 1.7 at the second biopsy (p < 0.001) and the staging index decreased significantly from 4.1 +/- 1.9 at the first biopsy to 2.7 +/- 2.4 at the second biopsy (p < 0.05). Macrophage infiltration and alpha-smooth muscle actin-positive cells in the glomerulus and interstitial region decreased significantly in post-therapy compared with pre-therapy. At the most recent follow-up, none of the 34 patients had renal insufficiency. CONCLUSIONS: Our study suggested that prednisolone and mizoribine therapy is effective for those patients with the risk of progression of IgAN.

Predicting the prognosis of renal dysfunction by renal expression of alpha-smooth muscle actin in children with MPGN type 1.

BACKGROUND: To clarify whether renal alpha-smooth muscle actin (alpha-SMA)-positive cell and macrophage accumulation can predict the prognosis of renal dysfunction, we evaluated them by means of multivariate analysis and compared them with other clinical predictors in patients with membranoproliferative glomerulonephritis (MPGN) type 1 with normal renal function. METHODS: We enrolled 35 patients with MPGN type 1 who had normal creatinine clearance at the time of renal biopsy. These patients were divided into 2 groups based on clinical status at the last examination. Group 1 consisted of 12 patients with normal urine and 12 patients with minor urinary abnormalities at the latest observation, whereas group 2 consisted of 7 patients with persistent nephropathy and 4 patients with renal insufficiency. The first and second renal biopsy findings, including alpha-SMA and CD68-positive staining, were investigated for both groups. RESULTS: Mean scores for glomerular and interstitial alpha-SMA staining in group 2 were significantly higher than those in group 1. At the second biopsy, mean scores for interstitial CD68-positive staining in group 2 were higher than those in group 1. Mean scores for glomerular and interstitial alpha-SMA at the first biopsy correlated with the chronicity index at the second biopsy in both groups. CONCLUSION: Our results suggest that glomerular and interstitial alpha-SMA expression at the first biopsy can predict the prognosis of children with MPGN type 1.

Expression pattern of glypican-1 mRNA after brain injury in mice.

Glypican-1, a heparan sulfate proteoglycan, is expressed in various tissues including developing and postnatal central nervous system. It serves as a receptor for heparin-binding molecules such as fibroblast growth factors (FGFs). We investigated whether glypican-1 was expressed after brain injury in adult mice. In situ hybridization study showed that glypican-1 mRNA was expressed in the region surrounding necrotic tissue, and that the signal intensity peaked 7 days after the cryo-injury. In addition, both FGF-2 and amyloid precursor protein (APP) were concurrently upregulated and colocalized with glypican-1 mRNA. Since FGF-2 and APP can bind to glypican-1 in vitro, the present study suggested that their autocrine/paracrine interactions with glypican-1 may be involved in neuronal regeneration and/or neurite-outgrowth inhibition after brain injury.