Tomoyuki Hotsubo

Role of Milk Whey in the Transmission of Human Cytomegalovirus Infection by Breast Milk

Breast‐fed infants are susceptible to human cytomegalovirus (HCMV) infection via breast milk. In our previous study, HCMV was isolated more frequently from breast milk at later than one month after delivery than from colostrum or early breast milk. To clarify the role of milk cells and whey in vertical infection by breast feeding, we separated breast milk into milk cells and whey and examined each fraction for the presence of HCMV. We collected breast milk from mothers who breast‐fed their infants (aged from 3 days to 2 months). The breast milk was centrifuged and separated into the middle layer (layer of milk whey) and the pellet (containing milk cells). We attempted to isolate HCMV from whey and to detect HCMV immediate early (IE) DNA in both milk whey and cells. HCMV was isolated from 7 out of 35 (20.0%) whey samples and HCMV IE DNA was detected from 15 out of 35 (42.9%) whey and/or milk cells. Detection rates of HCMV IE DNA in the whey layer and milk cells were 39.1% (25 out of 64) and 17.2% (11 out of 64), respectively. HCMV IE DNA was not detected in colostrum, but was detected in breast milk samples one month after delivery. Therefore, cell‐free HCMV shed into milk whey may have a more important role in vertical infection by breast milk than cell‐associated HCMV in the milk.

Comparison of an Immunochromatography Test with Multiplex Reverse Transcription-PCR for Rapid Diagnosis of Respiratory Syncytial Virus Infections

ABSTRACT A new commercial rapid 10-min one-step immunochromatography (IC) test, SAS RSV test, was compared to another IC test, Directigen EZ RSV, employing RT-PCR as the “gold standard” for detecting respiratory syncytial virus. Of 102 clinical samples, 79 were positive by RT-PCR, 66 (82.5%) were positive with the SAS RSV test, and 55 (69.6%) were positive with Directigen EZ RSV. The specificity of the new test was 91.3% (21 of 23), similar to that of Directigen EZ RSV (100% [23 of 23]). This test performs well enough to be used for patient care.

Detection of Human Cytomegalovirus DNA in Breast Milk by Means of Polymerase Chain Reaction

Three hundred and twenty‐five breast milk samples were examined for the occurrence of human cytomegalovirus (HCMV) by cell culture method. Virus was isolated from the milk in 1 of 177 samples collected within 6 days after delivery, 2 of 115 samples collected during the period of 7 days to 1 month after delivery, 10 of 33 samples collected over 1 month after delivery. Next, we tried to amplify HCMV DNA from the breast milk samples from HCMV seropositive mothers and seronegative mothers at 1 month after delivery by polymerase chain reaction. HCMV DNA was detected in 12 of 13 samples from seropositive mothers and in none of 7 samples from seronegative mothers. It was thought that all women seropositive for HCMV principally shed the virus into their breast milk at 1 month after delivery.

Three Novel IGSF1 Mutations in Four Japanese Patients With X-Linked Congenital Central Hypothyroidism

CONTEXT Congenital central hypothyroidism (C-CH) is a rare disease. We investigated the molecular basis of unexplained C-CH in 4 Japanese boys. PATIENTS AND METHODS C-CH was diagnosed by low free T4 and/or T3 and low basal TSH concentrations. We used whole-exome sequencing of one patient with C-CH to identify potential disease-causing mutations. Thereafter, PCR direct sequencing was performed to Identify genetic defects underlying C-CH in 3 more patients. We then assessed the effects of mutations identified in the Ig superfamily, member 1 (IGSF1), gene on protein expression and membrane trafficking. RESULTS All patients had congenital hypothyroidism, and 2 had definitive prolactin deficiency. Two patients were detected by neonatal screening. The other patients were diagnosed by short stature and failure to thrive. We identified a novel nonsense variant in IGSF1 by whole-exome sequencing in patient 1, which was confirmed by PCR direct sequencing (p.R1189X). PCR direct sequencing identified the identical nonsense mutation in patient 2. Patients 3 and 4 harbored distinct missense (p.V1082E) or nonsense (p.Q645X) mutations in IGSF1. The mothers of patients 1, 3, and 4 were heterozygous for these mutations. The R1189X mutant, which lacks the transmembrane domain, failed to traffic to the plasma membrane. V1082E could be observed at the cell surface, but at greatly diminished levels relative to the wild-type form of the protein. The severely truncated Q645X mutant could not be detected by Western blot. CONCLUSION Our findings provide additional genetic evidence that loss-of-function mutations in IGSF1 cause an X-linked form of C-CH and variable prolactin deficiency.

Rotavirus encephalitis and cerebellitis with reversible magnetic resonance signal changes

administration of a liquid valganciclovir formulation. Clin. Pharmacol. Ther. 2007; 81: 867–72. 15 Galli L, Novelli A, Chiappini E et al. Valganciclovir for congenital CMV infection: A pilot study on plasma concentration in newborns and infants. Pediatr. Infect. Dis. J. 2007; 26: 451–3. 16 Lombardi G, Garofoli F, Villani P et al. Oral valganciclovir treatment in newborns with symptomatic congenital cytomegalovirus infection. Eur. J. Clin. Microbiol. Infect. Dis. 2009; 28: 1465–70. 17 Nassetta L, Kimberlin D, Whitley R. Treatment of congenital cytomegalovirus infection: Implications for future therapeutic strategies. J. Antimicrob. Chemother. 2009; 63: 862–7.

Clinical characteristics and computed tomography findings in children with 2009 pandemic influenza A (H1N1) viral pneumonia

Abstract In this article we review the clinical characteristics and computed tomography (CT) findings in children with 2009 pandemic H1N1 influenza viral pneumonia. The medical charts of 88 children with pandemic H1N1 influenza virus infection, admitted to our hospital in Japan from 10 August to 28 December 2009, were reviewed; we compared the clinical features of these children with those of 61 children admitted with seasonal influenza A during the previous 3 seasons. Of 88 patients, 53 (60%) had radiographic findings consistent with pneumonia and 34 patients underwent a chest computed tomography (CT) scan. Pneumonia was a more frequent complication in children with pandemic H1N1 influenza compared with those with seasonal influenza (60% vs 11%; p < 0.001). The predominant CT findings were unilateral or bilateral multifocal consolidation (15/34; 44%) associated with ground-glass opacities in the peribronchovascular region. The second most common CT finding was unilateral diffuse consolidation or atelectasis in 1 or more lung zones (12/34; 35%). The chest CT findings of unilateral or bilateral multifocal consolidation often associated with ground-glass opacities were commonly seen in children with pandemic H1N1 influenza viral pneumonia. Atelectasis was seen in patients who required oxygen administration.

SPONASTRIME dysplasia: Report on a female patient with severe skeletal changes

We report on a 6-year-old girl with SPONASTRIME dysplasia, characterized by short-limbed dwarfism, a relatively large head, midfacial hypoplasia, a saddle nose, moderate deformities of the vertebral bodies, striated metaphyses, and normal intelligence. She showed severe skeletal changes including marked delay of epiphyseal ossification, evident metaphyseal dysplasia, and osteopathia striata more pronounced than in most of the previously reported patients with this disorder. The patient we describe and a male patient reported by Camera et al. [1994: Pediatr Radiol 24:322-324] are likely to represent the severely-affected end of the clinical spectrum of the disorder. These finding thus rule out the X-linked mode of inheritance of the disorder proposed by Camera et al. [1994: Pediatr Radiol 24: 322-324]. Alternatively, the two severely-affected patients may represent a variant form of the disorder. There is evidence that SPONASTRIME dysplasia is a genetically heterogeneous disorder.

Loss-of-Function and Gain-of-Function Mutations of Calcium-Sensing Receptor: Functional Analysis and the Effect of Allosteric Modulators NPS R-568 and NPS 2143

OBJECTIVE Activating mutations in the calcium-sensing receptor (CASR) gene cause autosomal dominant hypoparathyroidism, and heterozygous inactivating CASR mutations cause familial hypocalciuric hypercalcemia. Recently, there has been a focus on the use of allosteric modulators to restore the functional activity of mutant CASRs. In this study, the effect of allosteric modulators NPS R-568 and NPS 2143 on CASR mutants was studied in vitro. METHODS DNA sequence analysis of the CASR gene was undertaken in autosomal dominant hypoparathyroidism and familial hypocalciuric hypercalcemia Japanese patients, and the functional consequences for the Gi-MAPK pathway and cell surface expression of CASR were determined. Furthermore, we studied the effect of NPS R-568 and NPS 2143 on the signal transduction activity and cell surface expression of each mutant CASR. RESULTS We identified 3 activating mutations (S122C, P569H, and I839T) and 2 inactivating mutations (A110T and R172G) in patients. The activating and inactivating mutations caused leftward and rightward shifts, respectively, in the dose-response curves of the signaling pathway. NPS R-568 rescued the signal transduction capacity of 2 inactivating mutants without increasing cell surface expression levels. NPS 2143 suppressed the enhanced activity of the activating mutants without altering cell surface expression levels, although A843E, which is a constitutively active mutant, was suppressed to a lesser degree. CONCLUSIONS We have identified 4 novel mutations of CASR. Moreover, our results indicate that allosteric modulators can restore the activity of the loss- and gain-of-function mutant CASRs, identified in this study.

Prurigo Pigmentosa Disappeared Following Improvement of Diabetic Ketosis by Insulin

To the Editor: Prurigo pigmentosa is an inflammatory dermatosis characterized by pruritic erythematous papules and subsequent reticular pigmentation, the majority of which is distributed on the chest, back, and nape. Recently, a number of cases have been reported suggesting that ketosis may contribute to the pathogenesis of prurigo pigmentosa (1–5). We describe here a case of prurigo pigmentosa that was associated with the symptoms of diabetic ketosis and rapidly disappeared after the start of insulin therapy. A 15-year-old Japanese male visited a local physician with an eight-day history of pruritic eruptions over his shoulders, chest, back and upper arms. He also complained of a mild thirst and general fatigue that preceded by the appearance of the eruption by about a month and he had lost 6 kg in weight. Blood examination and urinalysis indicated diabetes mellitus, and he was referred to our hospital for further examination. His past history was unremarkable, and there was no history of diabetes mellitus in his family. Upon physical examination, erythema and papules were arranged in a reticulated pattern mainly on his back and chest (Fig. 1, left). Reticular pigmentation was also noted on the shoulders. Laboratory tests revealed type 1 diabetes mellitus and diabetic ketosis; urinary glucose, 3+; urinary ketones, 2+; blood glucose, 280 mg/dl; HbA1c, 14.7%; blood ketone bodies, 1,490 μmol/l; blood insulin, 3.5 μU/ml; blood C-peptide, 1.0 ng/ml; anti-glutamic acid decarboxylase, 22.1 U/ml. A skin biopsy of one of the erythematous papules on his back showed a perivascular mononuclear cell infiltration into the upper dermis. There was no obvious liquefaction in the basal layers, but a few melanophages and Civatte bodies were seen in the upper dermis. A diagnosis of prurigo pigmentosa with diabetic ketosis was made. Insulin therapy was started, and the eruption started to fade soon after the improvement of ketosis. Five days after starting insulin therapy, the eruption completely disappeared, leaving some reticular pigmentation (Fig. 1, right). In our case, the eruption of prurigo pigmentosa disappeared spontaneously following the improvement of ketosis by insulin therapy. Administration of minocycline or dapsone, which have been reported as effective treatments for prurigo pigmentosa (6), was not needed. According to a recent review of prurigo pigmentosa associated with diabetes mellitus, including those of the Japanese literature, 8 out of 13 cases of type 1 diabetic mellitus who received insulin therapy improved without any treatment for The Journal of Dermatology Vol. 30: 257–258, 2003

Treatment of Hypothyroidism due to Iodine Deficiency Using Daily Powdered Kelp in Patients Receiving Long-term Total Enteral Nutrition

We investigated thyroid function and urinary iodine concentration (UIC) in seven patients with severe motor intellectual disabilities. All seven received total enteral nutrition (TEN) for more than three years with a daily iodine intake of less than 20 µg. They were diagnosed as hypothyroidism due to iodine deficiency (HID) because of high TSH levels (7.6–82.3 µIU/ml), lower free T4 (FT4 0.4–1.5 ng/dl), negative anti-thyroid antibodies (anti-thyroglobulin antibody, anti-thyroidal peroxidase antibody) and extremely low UIC (<25–58 µg/l) levels. We gave them 1–2 g powdered kelp (200–400 µg as iodine) once a day, which restored their thyroid function and normalized their UICs. We proposed that daily powdered kelp would be effective and safe to treat HID in patient receiving long term TEN.