Screening for seemingly healthy newborns with congenital cytomegalovirus infection by quantitative real-time polymerase chain reaction using newborn urine: an observational study.

OBJECTIVE: Approximately 8-10% of newborns with asymptomatic congenital cytomegalovirus (cCMV) infection develop sensorineural hearing loss (SNHL). However, the relationship between CMV load, SNHL and central nervous system (CNS) damage in cCMV infection remains unclear. This study aimed to examine the relationship between urinary CMV load, SNHL and CNS damage in newborns with cCMV infection. STUDY DESIGN: The study included 23 368 newborns from two maternity hospitals in Saitama Prefecture, Japan. Urine screening for cCMV infection (quantitative real-time PCR) and newborn hearing screening (automated auditory brainstem response (AABR) testing) were conducted within 5 days of birth to examine the incidence of cCMV infection and SNHL, respectively. CNS damage was assessed by MRI of cCMV-infected newborns. RESULTS: The incidence of cCMV infection was 60/23 368 (0.257%; 95% CI 0.192% to 0.322%). The geometric mean urinary CMV DNA copy number in newborns with cCMV was 1.79×106 copies/mL (95% CI 7.97×105 to 4.02×106). AABR testing revealed abnormalities in 171 of the 22 229 (0.769%) newborns whose parents approved hearing screening. Of these 171 newborns, 22 had SNHL (12.9%), and 5 of these 22 were infected with cCMV (22.7%). Newborns with both cCMV and SNHL had a higher urinary CMV DNA copy number than newborns with cCMV without SNHL (p=0.036). MRI revealed CNS damage, including white matter abnormalities, in 83.0% of newborns with cCMV. Moreover, newborns with CNS damage had a significantly greater urinary CMV load than newborns without CNS damage (p=0.013). CONCLUSIONS: We determined the incidence of cCMV infection and urinary CMV DNA copy number in seemingly healthy newborns from two hospitals in Saitama Prefecture. SNHL and CNS damage were associated with urinary CMV DNA copy number. Quantification of urinary CMV load may effectively predict the incidence of late-onset SNHL and neurodevelopmental disorders.

Pitavastatin Reduces Inflammation in Atherosclerotic Plaques in Apolipoprotein E-Deficient Mice with Late Stage Renal Disease.

OBJECTIVES: Chronic renal disease (CRD) accelerates atherosclerosis and cardiovascular calcification. Statins reduce low-density lipoprotein-cholesterol levels in patients with CRD, however, the benefits of statins on cardiovascular disease in CRD remain unclear. This study has determined the effects of pitavastatin, the newest statin, on arterial inflammation and calcification in atherogenic mice with CRD. METHODS AND RESULTS: CRD was induced by 5/6 nephrectomy in cholesterol-fed apolipoprotein E-deficient mice. Mice were randomized into three groups: control mice, CRD mice, and CRD mice treated with pitavastatin. Ultrasonography showed that pitavastatin treatment significantly attenuated luminal stenosis in brachiocephalic arteries of CRD mice. Near-infrared molecular imaging and correlative Mac3 immunostaining demonstrated a significant reduction in macrophage accumulation in pitavastatin-treated CRD mice. Pitavastatin treatment reduced levels of osteopontin in plasma and atherosclerotic lesions in CRD mice, but did not produce a significant reduction in calcification in atherosclerotic plaques as assesses by histology. CRD mice had significantly higher levels of phosphate in plasma than did control mice, which did not change by pitavastatin. In vitro, pitavastatin suppressed the expression of osteopontin in peritoneal macrophages stimulated with phosphate or calcium/phosphate in concentrations similar to those found in human patients with CRD. CONCLUSION: Our study provides in vivo evidence that pitavastatin reduces inflammation within atherosclerotic lesions in CRD mice.

Cystathionine γ-lyase accelerates osteoclast differentiation: identification of a novel regulator of osteoclastogenesis by proteomic analysis.

OBJECTIVE: Clinical evidence has linked vascular calcification in advanced atherosclerotic plaques with overt cardiovascular disease and mortality. Bone resorbing monocyte-derived osteoclast-like cells are sparse in these plaques, indicating that their differentiation capability could be suppressed. Here, we seek to characterize the process of osteoclastogenesis by identifying novel regulators and pathways, with the aim of exploring possible strategies to reduce calcification. APPROACH AND RESULTS: We used a quantitative mass spectrometry strategy, tandem mass tagging, to quantify changes in the proteome of osteoclast-like cells differentiated from RAW264.7 cells in response to, receptor activator of nuclear factor κ-B ligand induction, a common in vitro model for osteogenesis. More than 4000 proteins were quantified, of which 138 were identified as novel osteoclast-related proteins. We selected 5 proteins for subsequent analysis (cystathionine γ-lyase [Cth/CSE], EGF-like repeat and discoidin I-like domain-containing protein 3, integrin α FG-GAP repeat containing 3, adseverin, and serpinb6b) and show that gene expression levels are also increased. Further analysis of the CSE transcript profile reveals an early onset of an mRNA increase. Silencing of CSE by siRNA and dl-propargylglycine, a CSE inhibitor, attenuated receptor activator of nuclear factor κ-B ligand-induced tartrate-resistant acid phosphatase type 5 activity and pit formation, suggesting that CSE is a potent inducer of calcium resorption. Moreover, knockdown of CSE suppressed expression of osteoclast differentiation markers. CONCLUSIONS: Our large-scale proteomics study identified novel candidate regulators or markers for osteoclastogenesis and demonstrated that CSE may act in early stages of osteoclastogenesis.

The oral administration of thermophile-fermented compost extract and its influence on stillbirths and growth rate of pre-weaning piglets.

Food produced via fermentation with mesophilic bacteria has been used to confer health benefits. In contrast, mammalian physiological responses to the intake of thermophile-fermented products have not been thoroughly investigated. We examined the effects of administering a compost extract consisting of fermented marine animals with thermophiles, including Bacillaceae, to pregnant sows and piglets. Retrospective studies were performed on two different swine farms (n=330-1050 sows). The rate of stillbirth was markedly lower in all parities of the compost extract-fed group compared to those of the control group (p≦0.001). Additionally, the birth to weaning period of newborns was significantly shorter (p<0.0001), while the ratio of weanlings per liveborn piglets was increased by more than 6.5% in the compost extract-fed group. Thus thermophiles and their products in the compost extract might promote growth and reduce stillbirths of piglets during the birth to weaning period.

Oral administration of multispecies microbial supplements to sows influences the composition of gut microbiota and fecal organic acids in their post-weaned piglets.

The timings of the administration of microbial supplements to control the populations of gut microbiota of piglets have been poorly understood. Here the effects of temporal administering multispecies microbial supplements to sows on the composition of gut microbiota and on the bacteria-mediated fecal metabolites in their offsprings were investigated. During gestation and lactation, pregnant sows were fed either a normal diet (group A) or a diet with multispecies supplements comprised of nine microbial species such as Lactobacillus delbrueckii subsp. bulgaricus, Bifidobacterium bifidum, Enterococcus faecium, Candida pintolopesii, and Aspergillus oryzae etc. (group B). All of the sows' piglets were temporarily fed with the same supplements around weaning in accordance with the guideline of the farm. This regimen was followed by a normal diet in both groups over one month thereafter. Under such conditions, the concentration of short-chain fatty acids (SCFAs) in fecal samples remarkably increased in group B compared to group A. When 16S rDNA sequences of the fecal bacteria were analyzed, the microbial structure of bacteria was different between both goups. Especially the Clostridium cluster IV and subcluster XIVa were particularly increased in group B, although the administered microbes were undetectable. Thus, temporal administration of multispecies-microbial supplements to pregnant sows changes the composition of SCFAs and gut microbiota in their offsprings.

Abrogation of bronchial eosinophilic inflammation and attenuated eotaxin content in protease-activated receptor 2-deficient mice.

Protease-activated receptor 2 (PAR2) belongs to the PAR family (PAR1 to PAR4), which is activated by serine proteases (trypsin, tryptase, etc.). In this study, we evaluated the role of PAR2 in allergic inflammation of airways using PAR2-deficient (PAR2(-/-)) mice. In wild- type mice, infiltration of eosinophils and high eotaxin content were found in bronchoalveolar lavage fluid (BALF) after ovalbumin (OA) sensitization and following challenge. In contrast, both OA-induced infiltration of eosinophils and increase of eotaxin content were abrogated in BALF from PAR2(-/-) mice. The activation of PAR2 might be essential in the production of eotaxin and consequential allergic inflammation in airways.

Brightly colored pigmentation in lower vertebrates: wonder searching its mechanisms and significance in the context of phylogeny.

This is a biographical sketch of my research and its related personal episodes with respect to brightly colored pigmentation in lower vertebrates. It includes a brief story of the studies on; (a) pterinosomes as a specific site of pteridine deposition in xanthophores or erythrophores of fish and amphibians, (b) a mosaic phenotype of chromatophores occurring in the reptiles and its implication for their developmental origin and differentiation mechanisms, (c) erythrophoroma as a tumor of erythrophores in goldfish, (d) the pluripotentials of erythrophoroma cells for expression of neural crest-derived characters in vitro, (e) pigment disorders occurring in hatchery-raised flounders and (f) recognition of pigment cell types by murine tyrosinase genes transfected into an orange-colored variant of medaka fish. Some of the personal affairs associated with the history of the Japanese community for pigment cell research were described to illustrate the background of these studies.

Effect of protease-activated receptor-2 deficiency on allergic dermatitis in the mouse ear.

To investigate the involvement of protease-activated receptor-2 (PAR-2) in allergic dermatitis, we generated PAR-2-deficient (PAR-2(-/-)) mice. Ear thickness, contact hypersensitivity (CH) induced by topical application of picryl chloride (PC) or oxazolone (Ox) after sensitization, and vascular permeability after ear passive cutaneous anaphylaxis (PCA) were compared between wild-type (WT) and PAR-2(-/-) mice. Ear thickness was almost the same in untreated WT and PAR-2(-/-) mice. Topical application of PC or Ox thickened the ears at 6, 24 and 48 h after challenge with a peak at 24 h in WT mice. In PAR-2(-/-) mice, the ear swelling induced by both PC and Ox was suppressed at every time point, and significant inhibition was found at 24 h in PC-induced CH and at 24 and 48 h in Ox-induced CH. Histopathological observation of the ears at 24 h after challenge revealed that PC- or Ox-induced ear edema and infiltration of inflammatory cells in WT mice were greatly attenuated in PAR-2(-/-) mice. The vascular permeability in the ears after PCA was not different between WT and PAR-2(-/-) mice. These results strongly suggest that PAR-2 plays a crucial role in type IV allergic dermatitis but not in type I allergic dermatitis.