The efficacy of the direct clinical intervention for infectious diseases by a pediatric infectious disease specialist in the pediatric ward of a tertiary medical facility without a pediatric antimicrobial stewardship program.

Antimicrobial stewardship programs (ASPs) have been introduced in most hospital complexes; however, they are not always useful for pediatric patients. The aim of this study is to investigate the efficacy of direct clinical intervention for infectious diseases by a pediatric infectious disease specialist in a tertiary medical facility without pediatric ASP. This retrospective study included 1,821 patients who were hospitalized in the pediatric ward of a large metropolitan hospital from 2010 to 2015. The clinical course, the use of intravenous antimicrobial agents and the results of a microbiological analysis were compared between the period after the beginning of direct intervention by the specialist (post-intervention period) and the previous period (pre-intervention period). In the post-intervention period, the proportion of the patients who received intravenous antimicrobial agents, the number of antimicrobial agents used for each episode, and the proportion of episodes in which an antimicrobial agent was re-administrated were significantly lower (P = 0.006, P = 0.004, P = 0.036, respectively), and the duration of antimicrobial treatment was significantly shorter (P < 0.001). In addition, narrower spectrum antimicrobial agents were used, and the incidence of meropenem-sensitive Pseudomonas aeruginosa significantly increased (P = 0.037) in the post-intervention period. There was no change of mortality between the two periods. Direct clinical intervention by a pediatric infectious diseases specialist is useful for the treatment of infectious diseases in the pediatric ward of a tertiary medical facility without a pediatric ASP. The creation of a pediatric ASP is recommended in hospital complexes.

BTK gene targeting by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector.

X-linked agammaglobulinemia (XLA) is one of the most common humoral immunodeficiencies, which is caused by mutations in Bruton's tyrosine kinase (BTK) gene. To examine the possibility of using gene therapy for XLA, we constructed a helper-dependent adenovirus/adeno-associated virus BTK targeting vector (HD-Ad.AAV BTK vector) composed of a genomic sequence containing BTK exons 6-19 and a green fluorescence protein-hygromycin cassette driven by a cytomegalovirus promoter. We first used NALM-6, a human male pre-B acute lymphoblastic leukemia cell line, as a recipient to measure the efficiency of gene targeting by homologous recombination. We identified 10 clones with the homologous recombination of the BTK gene among 107 hygromycin-resistant stable clones isolated from two independent experiments. We next used cord blood CD34⁺ cells as the recipient cells for the gene targeting. We isolated colonies grown in medium containing cytokines and hygromycin. We found that the targeting of the BTK gene occurred in four of the 755 hygromycin-resistant colonies. Importantly, the gene targeting was also observed in CD19⁺ lymphoid progenitor cells that were differentiated from the homologous recombinant CD34⁺ cells during growth in selection media. Our study shows the potential for the BTK gene therapy using the HD-Ad.AAV BTK vector via homologous recombination in hematopoietic stem cells.

Alagille-like syndrome with surprising karyotype: a case report.

BACKGROUND: Chromosome 5p partial monosomy (5p-syndrome) and chromosome 6p partial trisomy are chromosomal abnormalities that result in a variety of symptoms, but liver dysfunction is not normally one of them. Alagille syndrome (OMIM #118450) is a multisystem disorder that is defined clinically by hepatic bile duct paucity and cholestasis, in association with cardiac, skeletal, and ophthalmologic manifestations, and characteristic facial features. Alagille syndrome is caused by mutations in JAG1 on chromosome 20 or NOTCH2 on chromosome 1. Here, we report a preterm infant with karyotype 46,XX,der(5)t(5,6)(p15.2;p22.3) and hepatic dysfunction, who was diagnosed as having incomplete Alagille syndrome. CASE PRESENTATION: The Japanese infant was diagnosed based on the cardiac abnormalities, ocular abnormalities, characteristic facial features, and liver pathological findings. Analysis of the JAG1 and NOTCH sequences failed to detect any mutations in these genes. CONCLUSIONS: These results suggest that, besides the genes that are known to be responsible for Alagille syndrome, other genetic mutations also may cause Alagille syndrome.

Unique activation status of peripheral blood mononuclear cells at acute phase of Kawasaki disease.

Although Kawasaki disease (KD) is characterized by a marked activation of the immune system with elevations of serum proinflammatory cytokines and chemokines at acute phase, the major sources for these chemical mediators remain controversial. We analysed the activation status of peripheral blood mononuclear cells (PBMCs) by flow cytometry, DNA microarray and quantitative reverse transcription-polymerase chain reaction. The proportions of CD69+ cells in both natural killer cells and gammadeltaT cells at acute-phase KD were significantly higher than those at convalescent-phase KD. Microarray analysis revealed that five genes such as NAIP, IPAF, S100A9, FCGR1A and GCA up-regulated in acute-phase KD and the pathways involved in acute phase KD were related closely to the innate immune system. The relative expression levels of damage-associated molecular pattern molecule (DAMP) (S100A9 and S100A12) genes in PBMCs at acute-phase KD were significantly higher than those at convalescent-phase KD, while those of TNFA, IL1B and IL6 genes were not significantly different between KD patients and healthy controls. Intracellular production of tumour necrosis factor-alpha, interleukin-10 and interferon-gamma in PBMCs was not observed in KD patients. The present data have indicated that PBMCs showed a unique activation status with high expression of DAMP genes but low expression of proinflammatory cytokine genes, and that the innate immune system appears to play a role in the pathogenesis and pathophysiology of KD.

Identification of bacterial pathogens in pediatric community-acquired lower respiratory tract infection using a simplified procedure of sputum sampling and examination: comparison between hospitalized children with and without underlying diseases.

The aim of this study is to confirm the usefulness of sputum sampling from the hypopharynx through the nose to identify causative bacteria of pediatric community-acquired lower respiratory tract infection (CA-LRTI) and compare its features between the patients with and without underlying diseases. A retrospective study was performed on 244 pediatric patients hospitalized for CA-LRTI of suspected bacterial etiology. Sputum sample was obtained from these patients by aspirating airway secretion through the nose or the tracheostomy orifice, or coughing up by themselves. Sputum samples were assessed as suitable in 119 (74.4%) of 160 patients with CA-LRTI of suspected pure bacterial etiology. Ninety-six (70.1%) of 137 samples suctioned from the hypopharynx through the nose were suitable for bacterial examination. Seventy-eight (73.6%) of 106 patients identified with causative bacteria had some underlying diseases, and the other 28 patients did not have any underlying diseases. Proportions and antibiotics susceptibility profiles of the identified causative bacteria were almost similar in the patients with and without underlying diseases. Sputum sampling from the hypopharynx through the nose might be significant in pediatric CA-LRTI of suspected bacterial etiology. The initial treatment for patients without underlying diseases would be applicable to those with underlying diseases in the CA-LRTI of children.

Giant placental chorioangioma followed by circulatory failure of the newborn and infantile hemangioma: Case report.

Placental chorioangioma (CA) is a benign placental tumor. No specific treatment is required for asymptomatic cases. We report a female infant born to a mother with giant placental CA. However fetal growth was normal and, fetal hydrops was not detected by ultrasound examination until delivery, she had hydrops, subgaleal hematoma, thrombocytopenia, hemolytic anemia, respiratory distress and circulatory failure after birth. She was successfully treated without any neurological sequelae. At 2 months of age, infantile hemangioma appeared in her lower lip. The present case suggested that giant placental CA might cause postnatal problems and be associated with the development of infantile hemangioma.

A survey of the implementation status of selected infection control strategies in neonatal intensive care units in Japan.

BACKGROUND: Infection control strategies are implemented in all neonatal intensive care units (NICUs); however, the details of the strategies seem to differ among institutions. The purpose of this survey was to investigate the current implementation status of infection control strategies in NICUs in Japan and to identify and recommend appropriate strategies for the prevention of outbreaks in neonatal units. METHODS: This survey documented the current implementation status and methods of selected infection prevention and control measures (active surveillance cultures and standard precaution) in 453 Japanese NICUs/neonatal units registered with the Japan Society of Perinatal and Neonatal Medicine, using questionnaires, in May 2018. FINDINGS: The response rate was 48.1% (level I institutions, 25.5%; level II, 55.9%; level III, 64.2%). Surveillance cultures were performed every week and targeted all bacteria in most units. The proportion of level III institutions that experienced outbreaks over the previous five years was significantly higher than that of level II institutions (55% vs 27%, P=0.0003). However, wearing a mask was less frequently recommended in level III institutions (55.7%) than in level II institutions (67.9%). Meticillin-resistant Staphylococcus aureus (MRSA) was the most frequently reported bacterial pathogen responsible for NICU outbreaks. CONCLUSION: Infection prevention and control practices regarding active pathogen surveillance cultures and the use of barrier precautions varied widely in Japanese neonatal units. National guidelines and evidence-based recommendations are needed to rationalize and standardize current infection prevention and control practices in neonatal units in Japan.

High mobility group box 1 (HMGB1) and macrophage migration inhibitory factor (MIF) in Kawasaki disease.

OBJECTIVE: To investigate whether two proinflammatory cytokines, high mobility group box 1 (HMGB1) and macrophage migration inhibitory factor (MIF) are involved in the development of Kawasaki disease (KD). METHODS: Twenty-seven patients with KD were included in this study. Eleven patients with sepsis and 28 healthy children served as controls. Serum levels of HMGB1 and MIF were measured by corresponding enzyme-linked immunosorbent assay (ELISA) kits, respectively. Real-time polymerase chain reaction (PCR) was used to quantify the expression levels of genes encoding receptor for advanced glycation end-products (RAGE), an HMGB1 receptor, and CD74, an MIF receptor in peripheral blood mononuclear cells (PBMCs). RESULTS: Serum levels of HMGB1 and MIF in KD patients were the highest in the early acute phase and gradually decreased after defervescence. Serum HMGB1 and MIF levels in KD patients were significantly higher than those in controls (HMGB1, p<0.001; MIF, p<0.01). The expression levels of the RAGE gene and CD74 gene in KD patients were significantly higher than those in controls (RAGE, p<0.001; CD74, p<0.01). CONCLUSION: These data suggest that HMGB1 and MIF play an important role in immune responses in KD patients.

Activation of Nesfatin-1-Containing Neurones in the Hypothalamus and Brainstem by Peripheral Administration of Anorectic Hormones and Suppression of Feeding via Central Nesfatin-1 in Rats.

Peripheral anorectic hormones, such as glucagon-like peptide (GLP)-1, cholecystokinin (CCK)-8 and leptin, suppress food intake. The newly-identified anorectic neuropeptide, nesfatin-1, is synthesised in both peripheral tissues and the central nervous system, particularly by various nuclei in the hypothalamus and brainstem. In the present study, we examined the effects of i.p. administration of GLP-1 and CCK-8 and co-administrations of GLP-1 and leptin at subthreshold doses as confirmed by measurement of food intake, on nesfatin-1-immunoreactive (-IR) neurones in the hypothalamus and brainstem of rats by Fos immunohistochemistry. Intraperitoneal administration of GLP-1 (100 µg/kg) caused significant increases in the number of nesfatin-1-IR neurones expressing Fos-immunoreactivity in the supraoptic nucleus (SON), the area postrema (AP) and the nucleus tractus solitarii (NTS) but not in the paraventricular nucleus (PVN), the arcuate nucleus (ARC) or the lateral hypothalamic area (LHA). On the other hand, i.p. administration of CCK-8 (50 µg/kg) resulted in marked increases in the number of nesfatin-1-IR neurones expressing Fos-immunoreactivity in the SON, PVN, AP and NTS but not in the ARC or LHA. No differences in the percentage of nesfatin-1-IR neurones expressing Fos-immunoreactivity in the nuclei of the hypothalamus and brainstem were observed between rats treated with saline, GLP-1 (33 µg/kg) or leptin. However, co-administration of GLP-1 (33 µg/kg) and leptin resulted in significant increases in the number of nesfatin-1-IR neurones expressing Fos-immunoreactivity in the AP and the NTS. Furthermore, decreased food intake induced by GLP-1, CCK-8 and leptin was attenuated significantly by pretreatment with i.c.v. administration of antisense nesfatin-1. These results indicate that nesfatin-1-expressing neurones in the brainstem may play an important role in sensing peripheral levels of GLP-1 and leptin in addition to CCK-8, and also suppress food intake in rats.

Usefulness of electromagnetic flowmetry in intraoperative evaluation of aortic regurgitation associated with ventricular septal defect.

In 38 patients with aortic regurgitation (AR) associated with ventricular septal defect (VSD), indications for aortic valvuloplasty (AVP) or aortic valve replacement (AVR) were investigated by assessing AR ratio, which was measured by electromagnetic flowmetry of the ascending aorta during the operation. Residual AR was evaluated by auscultation postoperatively. Patients were assigned to group A if they underwent VSD closure alone and to group B if they underwent AVP in addition to VSD closure; patients who had no postoperative AR were assigned to group I and those with persistence of AR to group II. Thus, the patients were separated into 4 subgroups; A-I (9 patients), A-II (5 patients), B-I (20 patients) and B-II (4 patients). In subgroup A-I, mean AR ratio decreased from 9% to 7% postoperatively, in A-II from 19% to 16% (p less than 0.1), in B-I from 36% to 9% (p less than 0.01) and in B-II from 44% to 20% (p less than 0.05). An AR ratio of more than 25% should be regarded as an absolute indication for AVP or AVR. If the AR ratio is between 20 and 25%, the indication of AVP is determined by inspection of the aortic valve. A good correlation was found between AR ratio and the AR rate determined by the aortic angiographic findings (Sellers classification): grade I AR corresponded to an AR ratio of 5 to 15%, grade II to 20 to 35%, grade III to 25 to 50% and grade IV to 45% or higher (r = 0.87, p less than 0.01).

Marriage patterns among HTLV-I seropositive women in Japan.

A significantly higher percentage of asymptomatic HTLV-I seropositive pregnant women in the Kagoshima prefecture were married to men who were also born in that prefecture compared with seronegative women [138/166(83.1%), 221/306 (72.2%); P < 0.01]. A significantly higher percentage of the fathers of the seropositive women were born in the Kagoshima prefecture compared with the fathers of the seronegative women [152/166 (91.6%), 235/306 (76.8%); P < 0.01]. Additionally, a significantly higher seropositivity was found among pregnant women born in the Kagoshima prefecture who were married to men born in that prefecture compared with men born in other prefectures [5.8% (138/2374), 3.4% (28/819); P < 0.01]. Women born in other prefectures had a significantly lower seropositivity irrespective of the birthplace of their spouse [2.9% (12/418); P < 0.05, 3.0% (7/234)]. These findings indicate that HTLV-I seropositive women and their mothers chose their husbands from a smaller geographic region than seronegative women. This marriage pattern within an HTLV-I seropositive group may be one of the factors sustaining the present seroprevalence of HTLV-I.

Risk of HTLV-I infection in Japanese women who are last in birth order.

The percentage of last-born women among pregnant women who were seropositive for human T-lymphotropic virus type I (HTLV-I) significantly exceeded that among HTLV-I seronegative women (119/258 (46.1%): 89/251 (35.4%); P < 0.05). The findings suggest that last-born women are susceptible to HTLV-I infection. At least two possible interpretations of this birth-order effect are: (i) these last-born women were born to mothers who, on the average, were older than those of early-born women and, as a consequence, were more likely to have been seropositive and to have passed on HTLV-I to their daughters; (ii) husband-to-wife transmission of HTLV-I requires time to occur, so last-born women are more likely than early-born women to become infected.

Analysis of left ventricular motion after mitral valve replacement with a technique of preservation of all chordae tendineae. Comparison with conventional mitral valve replacement or mitral valve repair.

The postoperative regional left ventricular motion of 22 patients with a diagnosis of mitral regurgitation, and who underwent mitral valve replacement with preservation of chordae tendineae, were retrospectively analyzed by cineangiography in the early postoperative period and by multiple-gated cardiac blood pool scintigraphy in the mid-to-late postoperative period. The operation consisted of the division of the anterior leaflet into anterior and posterior segments, the shifting and reattachment of the divided segments to the mitral ring of the respective commissural areas, and the use of a low-profile bileaflet prosthetic valve. Control groups consisted of 28 patients with mitral regurgitation who underwent mitral valve replacement with a conventional technique and 16 patients who underwent mitral valve repair. Compared with the conventional mitral valve replacement group, the radial shortening of the left ventricle of the chordae-preserved mitral valve replacement group was greater at the apical septal, inferoapical, anterobasal, and anterolateral portions, whereas the radial shortening of the repair group was greater than that of the chordae-preserved group only at the inferolateral portion. The ejection fraction of the whole left ventricle was statistically greater in the chordae-preserved group, and also regional ejection fraction of the chordae-preserved mitral valve replacement group was greater at the apical septal, inferoapical, inferolateral, anterobasal, and anterolateral portions than that of the conventional mitral valve replacement group at these portions. On the other hand, the postoperative regional and global motion was identical to that of the mitral valve repair group except at the inferolateral portion. The result of this study supports a concept that maintenance of continuity between the mitral anulus and the papillary muscle has a beneficial effect on postoperative left ventricular performance.

Early and late results of reconstructive operation for congenital mitral regurgitation in pediatric age group.

Sixty-six children aged between 8 months and 15 years (average age 5.5 +/- 3.8 years) underwent mitral reconstructive operations for congenital mitral regurgitation between June 1969 and February 1987. The pathologic findings of the mitral valves were annular dilatation in 37 patients, cleft of the leaflet in eight, hypoplasia of the leaflet in 44, perforation of the leaflet in one, chordal elongation in 22, chordal absence in 16, and chordal shortening in seven. The methods of repair consisted of asymmetric annuloplasty (Kay-Reed method) in 61 patients, De Vega-type annuloplasty in one, plication of redundant leaflet in 15, and closure of cleft or perforation in nine. The single operative death (1.5%) was due to heart block. Follow-up data were available over 373.8 patient-years (average 5.7 years). The four late deaths (6.0%) were due to heart failure in two patients, pneumonia in one, and hepatitis in one. The actuarial survival rate was 93.1% +/- 3.1% at 7 years and 88.4% +/- 5.1% after 17 years. Valvuloplasty failed in 19 of the long-term survivors. One of these patients underwent mitral valve replacement 11 years after initial operation. The rate of freedom from reoperation was 86% +/- 10% after 17 years. The rate of freedom from valvuloplasty failure was 80% +/- 6.7% after 5 years, 67% +/- 7.2% after 10 years, and 44% +/- 11.9% after 15 years.

Long-term results of aortic valvuloplasty for aortic regurgitation associated with ventricular septal defect.

Long-term results in 64 patients who underwent aortic valvuloplasty for aortic regurgitation associated with ventricular septal defect are presented. The average age of the patients was 10.2 years and the average cardiothoracic ratio was 0.57 at the time of operation. The average degree of aortic regurgitation, as classified by Sellers, was 2.7. The type of ventricular septal defect was subpulmonic in 31 patients, perimembranous in 20, and was a total conal defect in five patients. Valvuloplasty was performed at one end of an aortic cusp in 23 patients, at two ends in 33, at three ends in six, and at four ends in two patients. There was one operative death (1.6%). The follow-up period was 447.7 patient-years, and there were no late deaths. The actuarial survival rate was 98.3% at 5, 10, and 15 years. Postoperative cardiac catheterization was performed in 40 patients. Valvuloplasty failure was defined as the presence of a cardiothoracic ratio greater than 0.6, a loud regurgitant murmur (Levine grade 3/6 or greater), moderate or severe aortic regurgitation (Sellers grade 3 or 4), or the necessity of reoperation. There were 17 patients whose valvuloplasty failed. The freedom from valvuloplasty failure was 74.2% at 5 and 10 years and 55.3% at 15 years. Eight patients underwent reoperation because of residual aortic regurgitation, and all survived. The freedom from reoperation was 90.1% at 5 years, 80.5% at 10 years, and 63.7% at 15 years. Multiple regression analysis revealed that older age, a greater cardiothoracic ratio, perimembranous ventricular septal defect, and multiple valvuloplasties were significant contributing factors for residual regurgitation after aortic valvuloplasty.

Echovirus type 9 epidemic in Kagoshima, southern Japan: seroepidemiology and clinical observation of aseptic meningitis.

An outbreak of aseptic meningitis caused by echovirus type 9 occurred between March and October, 1990, in Kagoshima, Southern Japan. Clinical manifestations and laboratory data of 259 children with aseptic meningitis who were admitted to the outpatient clinic of pediatrics in the Kagoshima City Hospital were analyzed (other diseases caused by echovirus type 9 were not investigated). The patients' age ranged from 1 month to 15 years and the highest incidence was in 4-year-old children. The male:female ratio was 1.3:1. Frequencies of headache (69%), vomiting (64%), neck stiffness (36%) and rash (195%) were lower than those in the previous reports in the United States or in the Europe. Pleocytosis in the cerebrospinal fluid increased with increasing age in the younger children. A predominance of neutrophils in cerebrospinal fluid lasted for 3 days or more after onset in 16% of the patients. Seroepidemiologic study suggested that the accumulation of susceptible children < 5 years of age had predisposed to the epidemic.

Exanthema subitum and human herpesvirus 6 infection: clinical observations in fifty-seven cases.

Exanthema subitum had been speculated to be a viral disease although its pathogen is unknown. Human herpesvirus 6 (HHV-6), first isolated in 1986, was proved by Yamanishi et al. to be the causal agent of exanthema subitum. To evaluate the role of HHV-6 as the causal agent in clinically diagnosed exanthema subitum, we tested for HHV-6 antibody in 57 infants with clinical exanthema subitum and exanthema subitum-like rash without fever. Of the 53 patients with clinical exanthema subitum 43 showed seroconversion or a significant rise in antibody titer to HHV-6, 7 were seropositive without significant rise and 3 remained seronegative. The clinical manifestations of these 43 infants with serologically confirmed HHV-6 infection were consistent with the classical characteristics of exanthema subitum. The 4 patients with atypical exanthema subitum showed significant rises in antibody titer. Our results therefore show that the majority of cases with typical clinical manifestations of exanthema subitum had HHV-6 infection. Most cases with HHV-6 infection had the typical clinical course of exanthema subitum, and a few cases might show an afebrile exanthema subitum-like rash.

Pharyngoconjunctival fever caused by adenovirus type 11.

Among a group of hospitalized children in Fukuoka, southern Japan, an epidemic of pharyngoconjunctival fever-like disease caused by adenovirus type 11 was observed in the autumn of 1988. Of the 47 children studied 38 were seronegative in neutralizing antibody for adenovirus type 11 before the epidemic, and seroconversion occurred in 16 (42%) including 5 subclinical cases. Of the 11 symptomatic patients the incidences of conjunctivitis, pharyngitis and fever were 91, 64 and 46%, respectively. Four patients (36%) had all three symptoms. Fifteen patients (94%) were boys. The sex predominance and high incidence of conjunctivitis suggested that infection may have been transmitted in the large bathroom where boys took baths together.

Trivalent cold recombinant influenza live vaccine in institutionalized children with bronchial asthma and patients with psychomotor retardation.

Twenty asthmatic children and 48 patients with severe psychomotor retardation were inoculated intranasally with trivalent cold-adapted recombinant (CR) influenza vaccine containing CR-125 (H1N1), CR-159 (H3N2) and CRB-117 (B). The vaccinees were mostly seropositive. Severe adverse reactions or asthmatic attacks were not observed, but 7 (15%) of 48 vaccinees with severe psychomotor retardation developed mild to moderate fever. Significant antibody responses in hemagglutination-inhibition tests were demonstrated in 33 (49%) vaccinees to CR-125, 20 (29%) to CR-159 and 8 (12%) to CRB-117. Two nosocomial outbreaks of influenza were observed in the subsequent winter. During an outbreak with H3N2 in one ward of severe psychomotor retardation patients, 2 (11%) of 18 vaccinees became infected compared with 10 (48%) of 21 placebo controls in the same ward (P < 0.05). In the other outbreak, with influenza B virus, 2 (14%) of 14 vaccinees and 13 (52%) of 25 controls in the ward for asthmatic children were infected (P < 0.05). The results indicate that trivalent CR vaccine is safe and effective against nosocomial outbreaks of influenza.

Propranolol for intractable hemolysis after open heart operation.

Postoperative intravascular hemolysis occurring in 2 patients was alleviated by propranolol. One patient underwent mitral valve replacement and had development of intractable hemolysis due to a paravalvular leak. The other patient underwent ventricular septal defect closure and had hemolysis caused by the Dacron patch. Both patients were given oral propranolol, and the degree of hemolysis decreased substantially. Although the exact mechanism of the propranolol effect on mechanical intravascular hemolysis is unclear, propranolol is thought to reduce the shearing stress between erythrocytes and the foreign material by slowing the velocity of the circulation.