Identification of Potential mRNA Biomarkers in Milk Small Extracellular Vesicles of Enzootic Bovine Leukosis Cattle

Enzootic bovine leukosis (EBL) is a disease caused by bovine leukemia virus (BLV); only a small percentage of BLV-infected cattle develop EBL and present with B-cell lymphosarcoma. There is no vaccine against BLV, treatment for EBL, or method for predicting the possibility of EBL onset, thus making EBL control difficult. Herein, to explore biomarkers for EBL in milk, we examined the mRNA profiles of small extracellular vesicles (sEVs) in milk from four BLV-uninfected and four EBL cattle by microarray analysis. It was revealed that 14 mRNAs were encapsulated in significantly higher quantities, and these mRNAs were therefore selected as biomarker candidates. Primers for these mRNAs were designed, and nine primer sets were available for quantitative real-time PCR. Nine mRNAs were evaluated for their availability as biomarkers for EBL using sEVs from newly-collected milk of 7 uninfected and 10 EBL cattle. The quantities of eight mRNAs (TMEM156, SRGN, CXCL8, DEFB4A, FABP5, LAPTM5, LGALS1, and VIM) were significantly higher in milk sEVs of EBL cattle than in those of uninfected cattle. Therefore, our findings indicate that these eight mRNAs in milk sEVs can be used as potential EBL biomarkers with combination use, although single mRNA use is not enough. Consequently, cattle at risk of EBL onset can be identified by monitoring the fluctuation in quantities of these mRNAs in milk before they develop EBL.     

Photodynamic Therapy in Macular Diseases of Asian Populations: When East Meets West

Purpose

We reviewed the indications, safety, and efficacy of photodynamic therapy with verteporfin in various macular diseases and vasculopathies, which are common in Asian populations, and compared the outcomes of photodynamic therapy in Asian patients with the outcomes in Caucasian patients.

Methods

Relevant clinical and laboratory original articles, case reports, and review articles that have been published in the literature between January 1999 and October 2004 were searched in Medline. The potential differences in the response to photodynamic therapy between Asian and Caucasian patients were evaluated. Articles in foreign languages with English abstracts were included.

Results

Macular diseases commonly seen in Asian populations, including choroidal neovascularization (CNV) of age-related macular degeneration, secondary to pathologic myopia or from an idiopathic cause, and choroidal vasculopathies such as central serous chorioretinopathy and polypoidal choroidal vasculopathy were included in the review. The results were tabulated and the differences with Caucasian populations were compared and highlighted.

Conclusion

Photodynamic therapy has been found to be an effective and noninvasive treatment for various subfoveal CNV and choroidal vasculopathies of the macula. Diverse behavior in different ethnic groups is observed. Jpn J Ophthalmol 2006;50:161–169 © Japanese Ophthalmological Society 2006     

Evaluation of the safety of xenon/bandpass light in vitrectomy using the A2E-laden RPE model

Background The purpose of the study was to investigate the brightness of the xenon/bandpass light in vitrectomy and assess its phototoxic effects using A2E-laden retinal pigment epithelial (RPE) cells. Methods The total luminous flux and spectral irradiance of 20- and 25-gauge endoilluminators connected to xenon lamps were measured and compared to those of 20- and 25-gauge endoilluminators connected to a halogen lamp. In vitro, A2E-laden cells were evenly exposed to xenon/bandpass light for 5 to 30 min positioned at 1 cm and 2 cm for a standard light probe and an implantable “chandelier” light probe, respectively, above the cells, and the cell viability was assessed using WST-1 assay. The cell viability was compared with cells exposed to 30 min of halogen light projected through a 20-gauge endoilluminator. Results The maximal total luminous flux of xenon/bandpass light emitted through the 20-gauge endoilluminator was 2.8 times higher than that of the halogen light. The total luminous flux of the 25-gauge endoilluminators was 0.6-1.1 times greater than the 20-gauge endoilluminators connected to the halogen light. The viability of the A2E-laden cells after exposure to the xenon/bandpass light was no different than that of the cells exposed to the halogen light when the total luminous flux of these lights was at the same level. Xenon/bandpass light from an implantable “chandelier” light probe induced A2E-mediated RPE damage to a similar extent as that of the halogen light through a 20-gauge endoilluminator. Conclusions A2E-mediated phototoxicity of xenon/bandpass light is comparable to that of halogen light.     

A dose-finding study of glycosylated G-CSF (Lenograstim) combined with CHOP therapy for stem cell mobilization in patients with non-Hodgkin's lymphoma

BACKGROUND: Peripheral blood stem cell (PBSC) reinfusion has been widely used for hematopoietic reconstitution after high-dose chemotherapy. However, the optimal dose of granulocyte colony-stimulating factor (G-CSF) for PBSC mobilization in combination with chemotherapy for autograft remains unknown. METHODS: To find the optimal dose of glycosylated G-CSF (lenograstim) for PBSC mobilization in combination with chemotherapy for aggressive non-Hodgkin's lymphoma (NHL), we conducted a dose-finding study on 43 newly diagnosed patients who had unfavorable prognostic factors. They received four to six courses of cyclophosphamide, doxorubicin, vincristine and prednisolone combined with lenograstim every 2 weeks (biweekly CHOP therapy). PBSC apheresis was started after the third course of biweekly CHOP therapy. Lenograstim was given daily from day 3 until the day of the last apheresis. The optimum dose of lenograstim was assessed based on mobilization efficacy and safety profiles at a daily single dose of 2, 5 and 10 microg/kg for eight patients in each level. RESULTS: The collected number of CD34+ cells in the first apheresis products was higher in the 5 microg/kg group than in the 2 microg/kg group (median, 4.22 x 10(6) vs 2.49 x 10(6) CD34+ cells/kg, P = 0.051). The highest dose of 10 microg/kg (median, 2.99 x 10(6) CD34+ cells/kg) failed to show a dose dependence in PBSC mobilization. The efficacy and safety of the 5 microg/kg dose were further confirmed in an additional 19 patients. CONCLUSIONS: The present study suggests that the recommended dose of lenograstim for PBSC mobilization with CHOP therapy in untreated NHL is 5 microg/kg.     

Neurotoxicity of petroleum benzine compared with n-hexane

Summary Petroleum benzine is one of the mixtures of organic solvents containing n-hexane. The occurrence of polyneuropathy in the workers using petroleum benzines is attributed mainly to n-hexane, though other hydrocarbons present are also suspected of having some neurotoxicity or some potential which could modify the neurotoxicity of n-hexane. The present experiment was performed in order to clarify the toxicity of petroleum benzine to the peripheral nerve and compare it with that of n-hexane.Forty rats were randomly divided into five groups. The groups were exposed to 200 ppm n-hexane, 500 ppm n-hexane, and petroleum benzine vapor containing 200 ppm n-hexane or 500 ppm n-hexane, together with aliphatic and aromatic hydrocarbons for 12 h a day for 24 weeks. The body weight, motor nerve conduction velocity, motor distal latency, and mixed nerve conduction velocities were measured before exposure and every 4 weeks of exposure. A rat from each exposed group was histopathologically examined after 24 weeks' exposure.The function of the peripheral nerve was conspicuously impaired by 500 ppm n-hexane, slightly impaired by 200 ppm n-hexane and petroleum benzine containing 500 ppm n-hexane, and even less impaired by petroleum benzine containing 200 ppm n-hexane. Degenerations of the myelin sheaths and axons were demonstrated in all exposed groups upon examination of the raveled tail nerves. Thus, the experiment revealed that petroleum benzine could impair the peripheral nerves, while some components of petroleum benzine were considered to inhibit the neurotoxicity of n-hexane.This investigation was partly supported by a grant for scientific research from the Chiyoda Mutual Life Foundation in 1980–1981     

Circulating immunocompetent cell profiles during oral cyclosporin therapy for immunoglobulin-resistant Kawasaki disease

Although the standard treatment for Kawasaki disease (KD)is intravenous immunoglobulin (IVIG) combined with oral aspirin,18％ of 1st IVIG is refractory [1].Recen...     

Non-traumatic renal arteriopelvic fistula

PURPOSE: In the present paper, we report on a 34-year-old female with macroscopic hematuria due to a nontraumatic renal arteriopelvic fistula (APF). The patient initially presented at another hospital with asymptomatic macroscopic hematuria. Following abdominal ultrasonography, computed tomography (CT) and laboratory data, no abnormal findings were seen. Therefore, the patient was referred to Teine Keijinkai Hospital for a more precise evaluation of the urinary tract and vascular abnormality. METHODS/RESULTS: Endoscopically, there was bleeding from the right ureteral orifice, so the patient was admitted for further examination. No abnormal findings were seen on urinary cytology and following an intravenous pyelogram. A selective right lower polar renal arteriogram revealed arterial extravasation directly into the pelvis before the venous phase, so APF of the kidney was diagnosed. The patient had no history of urinary tract trauma, so the APF was thought to be idiopathic. After transcatheter arterial embolization (TAE) with a gelatine sponge, macroscopic and microscopic hematuria disappeared and a low-density area was seen in the middle pole of the right kidney in an abdominal CT scan 4 days after TAE. This was thought to be renal infarction due to TAE. CONCLUSIONS: After discharge, the patient had no further hematuria.     

Correlation between water content and magnetization transfer ratio of the water component in bone marrow using gradient-echo imagings: normal case study

Objective. To determine the correlation between water content and magnetization transfer (MT) ratio of the water component in normal bone marrow using gradient-echo (GRE) magnetic resonance (MR) imaging. Design. Three types of GRE sequence – using fat suppression, using a combination of MT and fat suppression techniques, and without MT or fat suppression – were performed in nine healthy subjects. The water content of bone marrow assessed with GRE imaging was compared with that assessed with MR spectroscopy in three cases. The correlation between water content and MT ratio was observed using GRE imaging in nine subjects. Results and conclusions. The water content as assessed by GRE imaging and by MR spectroscopy were closely correlated (P<0.005, r=0.932). An inverse correlation between water content and MT ratio was observed in normal bone marrow (P<0.03, r=–0.432), which suggests that the water component in bone marrow includes free water protons.     

Investigation of mtDNA control region sequences in an Egyptian population sample

The sequences of mitochondrial DNA (mtDNA) control region were investigated in 101 unrelated individuals living in the northern region of Nile delta (Gharbia, N = 55 and Kafrelsheikh, N = 46). DNA was extracted from blood stained filter papers or buccal swabs. HV1, HV2 and HV3 were PCR amplified and sequenced; the resulted sequences were aligned and compared with revised Cambridge sequence (rCRS). The results revealed presence of total 93 different haplotypes, 86 of them are unique and 7 are shared haplotypes, the most common haplotype, was observed with a frequency, 2.97% of population sample. High mtDNA diversity was observed with genetic diversity and power of discrimination, 0.9982 and 0.9883, respectively. In this dataset the west Eurasian haplogroups predominated over the African haplogroups. The results would be useful for forensic examinations and human genetic studies.