Identification and characterization of sperm motility-initiating substance-2 gene in internally fertilizing Cynops species.

Sperm motility-initiating substance (SMIS) is an oviductal protein critical for internal fertilization in urodeles. It contributes to the establishment of various reproductive modes in amphibians and is thus a unique research model for the gene evolution of gamete-recognizing ligands that have diversified among animal species. In this study, a paralogous SMIS gene, smis2, was identified via the RNA sequencing of the oviduct of the newt, Cynops pyrrhogaster. The base sequence of the smis2 gene was homologous (˃90%) to that of the original smis gene (smis1), and deduced amino acid sequences of both genes conserved six cysteine residues essential for the cysteine knot motif. Furthermore, smis2 complementary DNA was identified in the oviduct of Cynops ensicauda, and the base substitution patterns also suggested that the smis gene was duplicated in the Salamandridae. Nonsynonymous/synonymous substitution ratios of smis1 and smis2 genes were 0.79 and 2.6, respectively, suggesting that smis2 gene evolution was independently driven by positive selection. Amino acid substitutions were concentrated in the cysteine knot motif of SMIS2. The smis2 gene was expressed in some organs in addition to the oviduct; in contrast, SMIS1 was only expressed in the oviduct. The SMIS2 protein was suggested to be produced and secreted at least in the oviduct and redundantly act in sperm. These results suggest that smis1 plays the original role in the oviduct, whereas smis2 may undergo neofunctionalization, which rarely occurs in gene evolution.

Acute retinal necrosis caused by herpes simplex virus type 2 in a 3-year-old Japanese boy.

Acute retinal necrosis (ARN), which is characterized by rapidly progressing peripheral retinal necrosis, is caused mainly by herpes simplex virus type 1, herpes simplex virus type 2 (HSV-2), or varicella-zoster virus. A previously healthy 3-year-old Japanese boy developed ARN in his left eye after being bruised by a milk container. HSV-2 DNA was detected in the aqueous humor of the affected eye. Serological testing suggested that the route of infection was from mother to child, although there was no past history of apparent HSV-2 infection. Childhood ARN has not been previously reported in Japan, possibly because of the low seroprevalence of HSV-2 in Japanese women. Pediatricians must be aware of this rare disease, which can affect individuals without a previous history of HSV even in a country with a low seroprevalence of HSV-2.

Bilateral pedicle stress fracture in a patient with osteoporotic compression fracture.

A case of bilateral pedicle stress fracture of L4 in a patient with osteoporotic compression fracture of L5 and without a history of major trauma or surgery is reported, and the literature is reviewed. Bilateral pedicle fracture is a rare entity and few cases have been reported in the literature. All reported cases had some underlying causative factors like previous spine surgery or stress related activities. To the best of the authors' knowledge, only one case of bilateral pedicle stress fracture without a history of trauma, previous spine surgery, or stress-related activities has been reported. A 77-year-old woman presented with severe low back pain and radiating pain in the right leg that was exacerbated after standing and walking. Plain radiograph showed pathological fracture at L5 level. Magnetic resonance imaging (MRI) revealed the compression of dural sac at L5 level. CT scan taken 3 months after admission revealed bilateral pedicle fractures through L4. The patient was treated with decompressive laminectomies of L4, followed by posterior spinal fusion with rigid pedicle screw fixation and autogenous bone graft mixed with hydroxyapatite. The patient achieved pain relief and returned to normal activity. Stress fracture of the pedicle within the proximal vertebra of an osteoporotic compression fracture of lumbar spine is an uncommon entity. It may, however, be an additional source of symptoms in patients with osteoporosis who present with further back pain. Surgeons caring for this group of patients should be aware of this condition.

Detection of cytomegalovirus DNA in preserved umbilical cords from patients with sensorineural hearing loss.

We performed a retrospective diagnostic study of congenital cytomegalovirus (CMV) infection in patients with sensorineural hearing loss (SNHL). CMV DNA in preserved umbilical cords was analyzed using real-time polymerase chain reaction analysis. Of 45 analyzable patients with SNHL, CMV DNA was detected in the preserved umbilical cords of 3 patients, all of whom had bilateral SNHL that lacked a clear onset period. CMV DNA was not detected in any of the patients with sudden SNHL or enlarged vestibular aqueduct-associated SNHL. The features of CMV-associated SNHL were more asymmetric than those of CMV-negative bilateral SNHL.

Posterior reversible encephalopathy syndrome in a child with bronchial asthma.

Although posterior reversible encephalopathy syndrome (PRES) is caused by various conditions, there have been no reports on PRES associated with bronchial asthma. We report a case with PRES during the treatment for severe asthmatic attack. A 4-year-old girl was treated for asthmatic attack with steroids. From the 10th hospital day, hypertension, pulmonary edema, and cardiomegaly were observed. In spite of treatment with furosemide, she became lethargic and had a generalized convulsion on the 23rd hospital day. CT showed low density areas in the bilateral occipital white matter and MRI on the 28th hospital day demonstrated high intensity areas in the same regions on T2-weighted and FLAIR images. After discontinuation of corticosteroid and further antihypertensive therapy, her consciousness improved. MRI on the 67th hospital day had no abnormalities and no neurological sequelae were seen at 2 years after the event. We should be aware that PRES is a rare but important adverse event related to steroid therapy, because hypertension and water retention are major adverse effects of steroids.

Relationship between human cytomegalovirus glycoprotein B genotype and serum alanine aminotransferase elevation in infants.

The glycoprotein B (gB) region of the human cytomegalovirus (HCMV) is a major envelope glycoprotein that is a principal target of neutralizing antibodies and is known to stimulate the immune response of cytotoxic T lymphocytes. HCMV is currently classified into four genotypes on the basis of the nucleotide sequence of the gB region. The presence of HCMV in patients under 3 years of age was determined by subjecting urine samples taken from the patients to polymerase chain reaction (PCR) analysis. Analysis by direct sequencing of the gB region was carried out in 90 cases. These cases were grouped into the gB genotype 1 and gB genotype 3. Of 28 cases with a peak serum alanine aminotransferase (ALT) level(> or =100 IU/l), the duration of observed serum ALT elevation in the gB genotype 1 patients (166.7+/-126.7 days [mean+/-S.D.] [19 cases]) was significantly longer than that in the gB genotype 3 patients (39.7+/-31.7 days [9 cases]) (p<0.01). In the 54 cases with a serum ALT level(> or =50 IU/l), similar tendency was seen (p<0.05). These findings suggest that when serum ALT elevation is confirmed in young children infected with HCMV, analysis of the gB region is helpful for prediction of the duration of serum ALT elevation in the early stage of infection.

Comparison of real-time and nested PCR assays for detection of herpes simplex virus DNA.

We performed a real-time PCR assay to detect herpes simplex virus (HSV) DNA, and compared it prospectively with a nested PCR assay in 164 clinical samples (109 cerebrospinal fluid and 55 sera) from patients suspected of having neonatal HSV infection or HSV encephalitis. In 25 of 164 samples, HSV DNA was detected by the nested PCR assay. All samples positive for HSV DNA in the nested PCR assay were also positive in the real-time PCR assay, and all but two samples negative for HSV DNA in the nested assay were negative in the real-time assay. The real-time PCR assay thus had a sensitivity of 100% and a specificity of 99%, when compared with the nested assay. Sequential assays in a case of disseminated HSV showed that a decrease in HSV DNA paralleled clinical improvement. Quantification of HSV DNA by real-time PCR was useful for diagnosing and monitoring patients with HSV encephalitis and neonatal HSV infection.

Quantitation of viral load in neonatal herpes simplex virus infection and comparison between type 1 and type 2.

Neonatal herpes simplex virus (HSV) infection is a severe disease with high mortality and morbidity in spite of the development of effective anti-viral therapies. The viral load in neonatal herpes simplex virus (HSV) infection was measured retrospectively in 37 patients. HSV DNA copy numbers in serum and cerebrospinal fluid (CSF) were quantified using a real-time PCR assay. Patients with disseminated infection had a higher viral load in their sera. whereas patients with central nervous system (CNS) infection exhibited a higher viral load in the CSF. The viral load was significantly higher in the serum of patients who died later. Interestingly, patients with HSV type-2 infection exhibited more CNS involvement and neurological impairment, together with a high viral load in the CSF, than did HSV type-1 patients. These results suggest that quantitation of HSV viral load may be useful for assessing the prognosis, and may provide additional information for the management of neonatal HSV infection.

Evaluation of systemic inflammatory responses in neonates with herpes simplex virus infection.

Neonatal herpes simplex virus (HSV) infection is a severe disease with high mortality and morbidity. To investigate the pathogenesis of neonatal HSV infection, we examined inflammatory responses and markers of apoptosis in patients with neonatal HSV infection. Concentrations of inflammatory cytokines and markers of apoptosis were significantly higher in patients with disseminated HSV infection and were correlated with HSV load. It appears that the immunopathological damage that results from host responses to viral infection leads to organ dysfunction in patients with neonatal HSV infection.