Clinical and genetic investigation of a Japanese family with cardiac fabry disease. Identification of a novel α-galactosidase A missense mutation (G195V).

Fabry disease is an X-linked lysosomal storage disorder caused by mutations of the α-galactosidase A gene (GLA), and the disease is a relatively prevalent cause of left ventricular hypertrophy mimicking idiopathic hypertrophic cardiomyopathy. We assessed clinically 5 patients of a three-generation family and also searched for GLA mutations in 10 family members. The proband had left ventricular hypertrophy with localized thinning in the basal posterior wall and late gadolinium enhancement (LGE) in the near-circumferential wall in cardiovascular magnetic resonance images and her sister had vasospastic angina pectoris without organic stenosis of the coronary arteries. LGE notably appeared in parallel with decreased α-galactosidase A activity and increased NT-pro BNP in our patients. We detected a new GLA missense mutation (G195V) in exon 4, resulting in a glycine-to-valine substitution. Of the 10 family members, 5 family members each were positive and negative for this mutation. These new data extend our clinical and molecular knowledge of GLA gene mutations and confirm that a novel missense mutation in the GLA gene is important not only for a precise diagnosis of heterozygous status, but also for confirming relatives who are negative for this mutation.

Rapid and sensitive detection of norovirus genomes in oysters by a two-step isothermal amplification assay system combining nucleic acid sequence-based amplification and reverse transcription-loop-mediated isothermal amplification assays.

We developed a two-step isothermal amplification assay system, which achieved the detection of norovirus (NoV) genomes in oysters with a sensitivity similar to that of reverse transcription-seminested PCR. The time taken for the amplification of NoV genomes from RNA extracts was shortened to about 3 h.

Periorbital cyst with bone defect in a dog.

A 4-year-old female Miniature Dachshund was referred with a chief complaint of right periorbital swelling that had not responded to antibiotic therapy. Ultrasonography and fine-needle aspiration revealed that the periorbital lesion had a cystic structure without any inflammatory or neoplastic cells. Computed tomography (CT) showed that the cyst occupied a defect in the periorbital maxillary, lacrimal, and frontal bones and had invaded the nasal cavity. The lesion was histologically suspected by incisional biopsy as an epithelial cyst.

Contrast harmonic imaging of canine hepatic tumors.

Six adult healthy Beagles were used to investigate the hepatic perfusion dynamics of Levovist, a contrast agent used in contrast harmonic imaging (CHI). In addition, 8 dogs with hepatocellular carcinoma (HCC) and 2 dogs with metastatic hepatic hemangiosarcoma (HSA) were used to characterize both the CHI findings with Levovist. In the Beagles, the start of intravenously injected Levovist into the aorta between the cranial mesenteric and renal arteries and the portal vein at the hepatic hilum were 5.47 +/- 1.52 sec and 16.03 +/- 3.39 sec, respectively. As a characteristic CHI finding in the 8 dogs with HCC, the early arterial phase showed a fine network of blood flow enhanced at the surrounding region and within the tumor in all the 8 dogs (100%), and the post vascular phase demonstrated a defect in the whole tumor and an enhancement of the surrounding hepatic tissues in 7 dogs (87.5%). In the 2 dogs with HSA, characteristic finding in which the early arterial and late vascular phases showed a rim contrast enhancement pattern, and the post vascular phase revealed that the whole tumor lacked contrast enhancement and the surrounding hepatic tissues was clearly enhanced. In dogs, the start of the early arterial and late vascular phases, and the characterizations of the CHI findings in HCC and HSA were suggested to be similar to those in humans. Therefore, CHI is thought to be useful for the diagnosis of HCC and metastatic hepatic HSA in dogs as well as in humans.

Fibrodysplasia ossificans progressiva-like condition in a cat.

Fibrodysplasia ossificans progressiva (FOP)-like condition was diagnosed in a Japanese domestic cat with stiffness, marked atrophy of the muscles, and limited mobility of all joints in both the pelvic limbs. Etretinate, a retinoid, was used for medical management; however, no improvement in the clinical signs was observed. Inheritance of the disorder has not yet been demonstrated. Furthermore, the clinical signs and histopathological findings of feline FOP-like condition in the present case differed from those of the previously reported cases.

Assessment of left ventricular function using pulsed tissue Doppler imaging in healthy dogs and dogs with spontaneous mitral regurgitation.

Pulsed tissue Doppler imaging (pulsed TDI) has been demonstrated to be useful for the estimation of left ventricular (LV) systolic and diastolic functions in various human cardiac diseases. The objectives of this study were to investigate the relationship between pulsed TDI and LV function by using cardiac catheterization in healthy dogs and to evaluate the clinical usefulness of pulsed TDI in dogs with spontaneous mitral regurgitation (MR). The peak early diastolic velocity (E'), peak atrial systolic velocity (A'), and peak systolic velocity (S') were detectable in the velocity profiles of the mitral annulus in all the dogs. In the healthy dogs, S' and E' were correlated with LV peak +dP/dt and -dP/dt, respectively. E' was lower in dogs with MR than in dogs without cardiac diseases. E/E' in the MR dogs with decompensated heart failure was significantly increased in comparison with those with compensated heart failure. The sensitivity and specificity of the E/E' cutoff value of 13.0 for identifying decompensated heart failure were 80% and 83%, respectively. In addition, E/E' was significantly correlated with the ratio of left atrial to aortic diameter. These findings suggest that canine pulsed TDI can be applied clinically for estimation of cardiac function and detection of cardiac decompensation and left atrial volume overload in dogs with MR.

Successful treatment by percutaneous transvenous coil embolization in a small-breed dog with intrahepatic portosystemic shunt.

A Miniature Dachshund, 3-month-old, 3.1 kg, was diagnosed as an intrahepatic portosystemic shunt (PSS) with the shunting vessel in 6-mm diameter. Percutaneous transvenous coil embolization (PTCE) was performed with a stainless steel coil in 8-mm diameter. Intraoperative portal pressure elevated about 2.5 times after one-stage coil occlusion. Two weeks after the PTCE, serum bile acid levels reduced within the normal range. The portogram showed complete occlusion of the shunting vessel 4 months after the PTCE. Approximately 3 years after the PTCE, the patient has shown no clinical signs. PTCE could be performed more easily and less invasively in a small-breed dog. It is therefore suggested that PTCE is a promising therapeutic technique in canine intrahepatic PSS.

Role of NKp46 expression in cytokine production by CD56-positive NK cells in the peripheral blood and the uterine endometrium.

PROBLEM: To investigate the role of natural cytotoxicity receptor, NKp46 expression in cytokine-producing NK cells. METHOD OF STUDY: CD56(+) /NKp46(+) NK cells from the peripheral blood and the uterine endometrium were magnetically separated. IFN-γ, TNF-α, IL-4, IL-10, and TGF-ß(1) expressions on NK cells were investigated using multicolor flow cytometry. RESULTS: Peripheral blood and uterine endometrial NK cells were grouped into 4 subpopulations based upon the degree of CD56 and NKp46 expressions. NKp46 expression was associated with higher frequency of cytokine-producing NK cells, including CD56(dim) NK cells. The percentage of TNF-α(+) and IL-10(+) NK cells per total CD56(+) /NKp46(+) NK cells in the uterine endometrium showed a significant correlation with those of the peripheral blood in all subpopulations, but that of IFN-γ(+) , IL-4(+,) and TGF-ß(1) (+) NK cells showed partial correlation. CONCLUSION: Expression of NKp46 is involved in cytokine production of CD56(+) NK cells in the peripheral blood and the uterine endometrium.

Rapid diagnosis of adenovirus respiratory tract infections using the chromatographic immunoassay test in the pediatric outpatient setting.

To assess the usefulness of a new rapid chromatographic immunoassay test for the detection of adenovirus, a prospective 3-year study was conducted in 587 febrile outpatient children suspected of adenovirus infection. A total of 332 children were diagnosed with this infection, using a viral culture. The sensitivity and specificity of the rapid test were 89.2% and 98.0%, respectively.

Recombinant norovirus implicated in gastroenteritis outbreaks in Hiroshima Prefecture, Japan.

Norovirus (NoV) is a major etiological agent of acute gastroenteritis outbreaks worldwide. A total of 314 fecal specimens collected from patients of 39 NoV gastroenteritis outbreaks in Hiroshima Prefecture, Japan, between December 2001 and April 2006 were tested for the occurrence of recombinant NoVs. Sixteen genotypes (GI/1, GI/2, GI/4, GI/7, GI/8, GI/11, GI/14, GII/2, GII/3, GII/4, GII/5, GII/6, GII/8, GII/12, GII/14, and GII/untypeable) were detected in the 39 outbreaks based on capsid sequences and GII/4 was predominant recently. Twelve strains detected in 11 (28.2%) of the 39 outbreaks were suspected to be recombinants by using Simplot and Recco analyses and five recombinant genotypes, GII/4-GII/12 (five strains), GIIb-GII/3 (four strains), GII/4-GII/2 (one strain), GII/4-GII/14 (one strain), and GI/2-GI/8 (one strain), were identified based on RNA-dependent RNA polymerase and capsid sequences. None of the strains genotyped as GII/4 based on the capsid sequence was identified as a recombinant. The putative recombination points in the recombinant strains were placed either upstream or downstream of the open reading frame (ORF) 1 and ORF2 overlap. The present study indicates the following: (a) recombination among ORFs is common in nature, (b) the involvement of recombinant NoVs in gastroenteritis outbreaks is extensive even in a local area such as Hiroshima Prefecture, Japan, and (c) the conserved region (ORF1 and ORF2 overlap) has a meaningful function against the recombination event.

Simultaneous detection and genogroup-screening test for norovirus genogroups I and II from fecal specimens in single tube by reverse transcription- loop-mediated isothermal amplification assay.

We developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the rapid detection of noroviruses (NoVs) in a genogroup-specific manner in a previous study. In this study, to detect NoVs more easily and simply, we have developed an RT-LAMP assay for the simultaneous detection of NoV genogroup I (GI) and II (GII) genomes in a single tube. The genogrouping was achieved by using fluorescence-labeled primers, and the green and red colors for GI and GII, respectively, sometimes with yellow color for GI and GII mixture, were indicated on the agarose gel under UV light at 312 nm.