Clinical relevance of quantified fundus autofluorescence in diabetic macular oedema.

PURPOSE: To quantify the signal intensity of fundus autofluorescence (FAF) and evaluate its association with visual function and optical coherence tomography (OCT) findings in diabetic macular oedema (DMO). METHODS: We reviewed 103 eyes of 78 patients with DMO and 30 eyes of 22 patients without DMO. FAF images were acquired using Heidelberg Retina Angiograph 2, and the signal levels of FAF in the individual subfields of the Early Treatment Diabetic Retinopathy Study grid were measured. We evaluated the association between quantified FAF and the logMAR VA and OCT findings. RESULTS: One hundred and three eyes with DMO had lower FAF signal intensity levels in the parafoveal subfields compared with 30 eyes without DMO. The autofluorescence intensity in the parafoveal subfields was associated negatively with logMAR VA and the retinal thickness in the corresponding subfields. The autofluorescence levels in the parafoveal subfield, except the nasal subfield, were lower in eyes with autofluorescent cystoid spaces in the corresponding subfield than in those without autofluorescent cystoid spaces. The autofluorescence level in the central subfield was related to foveal cystoid spaces but not logMAR VA or retinal thickness in the corresponding area. CONCLUSIONS: Quantified FAF in the parafovea has diagnostic significance and is clinically relevant in DMO.

Association between cystoid spaces on indocyanine green hyperfluorescence and optical coherence tomography after vitrectomy for diabetic macular oedema.

PURPOSE: To study retrospectively the characteristics of residual indocyanine green (ICG) fluorescence after ICG-assisted vitrectomy and the association with spectral-domain optical coherence tomography (SD-OCT) findings in diabetic macular oedema (DMO). METHODS: Thirteen consecutive eyes of 12 patients for whom fundus near-infrared fluorescence and 20° retinal sectional images were obtained using HRA2 and Spectralis OCT, respectively, 5 days after vitrectomy combined with ICG-assisted inner limiting membrane peeling for DMO. The relationship between the characteristics of the ICG hyperfluorescence and the cystoid spaces in the outer plexiform layer (OPL) on SD-OCT images was evaluated. RESULTS: A total of 390 well-demarcated areas of ICG hyperfluorescence were delineated on 20° radial OCT scans dissecting the fovea 5 days after vitrectomy. The areas of ICG hyperfluorescence in the parafovea or perifovea were significantly smaller than those at the fovea. Most areas of hyperfluorescence were irregularly shaped in the parafovea and perifovea, whereas 18 of 38 areas of hyperfluorescence were round or oval at the fovea (P<0.001). SD-OCT delineated the cystoid spaces in the OPL in 73 areas of hyperfluorescence that were round or oval and accompanied by dark spots more frequently than that without cystoid spaces on OCT images (P<0.001 and P=0.002). Of the 123 cystoid spaces in the OPL on OCT images, 44 did not have ICG hyperfluorescence, had lower OCT reflectivity, and contained fewer hyperreflective foci than those with ICG hyperfluorescence (P<0.001 and P=0.020). CONCLUSION: The results provided novel interpretations of the ICG hyperfluorescence and its association with OCT characteristics of the cystoid spaces in DMO.

Meropenem as predictive risk factor for isolation of multidrug-resistant Pseudomonas aeruginosa.

The objective of this study was to explore independent risk factors for the isolation of multidrug-resistant (MDR) Pseudomonas aeruginosa in a Japanese university hospital between January 1997 and December 2010. MDR P. aeruginosa was defined when the organism was resistant or intermediately susceptible to all five antimicrobials tested. In all, 159 patients with MDR P. aeruginosa were identified over the 14-year period. Multivariate logistic regression analysis revealed that prolonged hospital stay, prior exposure to meropenem and fluoroquinolones, and patients suffering from diabetes mellitus or receiving surgery were predictive risk factors for the isolation of MDR P. aeruginosa.

Use of a dual priming oligonucleotide system to detect multiple sexually transmitted pathogens in clinical specimens.

AIMS: To evaluate a new dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) assay for detection of six sexually transmitted pathogens, including Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum and Trichomonas vaginalis. METHODS AND RESULTS: Using 130 clinical specimens, the results obtained by the multiplex PCR, previously established in-house PCR and COBAS Amplicor PCR assays were compared. The specimens frequently contained multiple pathogens (34/130 specimens). The multiplex PCR assay had an overall sensitivity of 96% and specificity of 100% compared to the in-house PCR assay at >20 microg ml(-1) of DNA concentrations in samples and there was no cross-reaction with nonpathogenic Neisseria species that cause the majority of false-positive results with the COBAS Amplicor PCR assay. CONCLUSIONS: The DPO-based multiplex PCR assay detected the six sexually transmitted pathogens in clinical specimens with a high sensitivity and specificity, although its sensitivity was dependent on the DNA content of the samples. SIGNIFICANCE AND IMPACT OF THE STUDY: It is the first report about the new DPO-based technique to detect multiple sexually transmitted pathogens in a single assay, which has considerable potential to diagnose the infections accurately and rapidly.

Cloning and expression analysis of YY1AP-related protein in the rat brain.

YY1AP-related protein (YARP) is a structural homolog of YY1AP, a transcriptional coactivator of the multifunctional transcription factor YY1. We cloned a rat YARP cDNA that encoded a 2256 amino acid protein with 93% homology to the human counterpart. Northern blots revealed significant expression of the YARP gene in the rat brain. In situ hybridization demonstrated its expression in neurons throughout the brain, including pyramidal cells in the cerebral cortex and hippocampus and granule cells in the dentate gyrus. YARP was coexpressed with YY1 in these same neuronal cells. However, there was no evidence of YARP expression in glia. In the developing rat brain, the level of YARP mRNA ( approximately 10 kb) peaked at embryonic day 18 and promptly declined thereafter to reach the steady-state level found in adulthood, by 14 days after birth. These results suggest that YARP functions at a late stage of neurogenesis during perinatal development of the rat brain, as well as in mature neurons.

Molecular cloning of a structural homolog of YY1AP, a coactivator of the multifunctional transcription factor YY1.

YY1 is a multifunctional transcription factor that activates or represses gene transcription depending on interactions with other regulatory proteins that include coactivator YY1AP. Here, we describe the cloning of a novel homolog of YY1AP, referred to as YARP, from the human neuroblastoma cell line SK-N-SH. The cloned cDNA encoded a 2240 amino acid protein that contained a domain which was 97% homologous to an entire YY1AP sequence of 739 amino acids. Two splice variants, YARP2 and YARP3, were also cloned. Northern blotting demonstrated the YARP mRNA (approximately 10 kb), which was increased 1.7-fold after dibutyryl cAMP-induced neural differentiation of the cells. Presence of YARP mRNA was also confirmed in human tissues such as the heart, brain and placenta. Bioinformatic analysis predicted various functional motifs in the YARP structure, including nuclear localization signals and domains associated with protein-protein interactions (PAH2), DNA-binding (SANT), and chromatin assembly (nucleoplasmin-like), outside the YY1AP-homology domain. Thus, we propose that YARP is multifunctional and plays not only a role analogous to YY1AP, but also its own specific roles in DNA-utilizing processes such as transcription.

Genomic imprinting in Dicer1-hypomorphic mice.

To address the function of RNA interference (RNAi) in transcriptional silencing in mammals, we analyzed genomic imprinting in Dicer1-hypomorphic mice, in which Dicer1 expression was significantly reduced. We did not observe any abnormality in the allelic expression of imprinted genes in these mice or their offspring, suggesting that reduced expression of Dicer1 did not significantly affect the maintenance and reprogramming of imprinting.

Efficacy of gemtuzumab ozogamicin on ATRA- and arsenic-resistant acute promyelocytic leukemia (APL) cells.

Acute promyelocytic leukemia (APL) cells express a considerable level of CD33, which is a target of gemtuzumab ozogamicin (GO), and a significantly lower level of P-glycoprotein (P-gp). In this study, we examined whether GO was effective on all-trans retinoic acid (ATRA)- or arsenic trioxide (ATO)-resistant APL cells. Cells used were an APL cell line in which P-gp was undetectable (NB4), ATRA-resistant NB4 (NB4/RA), NB4 and NB4/RA that had been transfected with MDR-1 cDNA (NB4/MDR and NB4/RA/MDR, respectively), ATO-resistant NB4 (NB4/As) and blast cells from eight patients with clinically ATRA-resistant APL including two patients with ATRA- and ATO-resistant APL. The efficacy of GO was analyzed by (3)H-thymidine incorporation, the dye exclusion test and cell cycle distribution. GO suppressed the growth of NB4, NB4/RA and NB4/As cells in a dose-dependent manner. GO increased the percentage of hypodiploid cells significantly in NB4, NB4/RA and NB4/As cells, and by a limited degree in NB4/MDR and NB4/RA/MDR cells. Similar results were obtained using blast cells from the patients with APL. GO is effective against ATRA- or ATO-resistant APL cells that do not express P-gp, and the mechanism of resistance to GO is not related to the mechanism of resistance to ATRA or ATO in APL cells. Leukemia (2005) 19, 1306-1311. doi:10.1038/sj.leu.2403807; published online 26 May 2005.

[Acute pulmonary embolism after cesarean section; report of a case].

We present a case of acute pulmonary embolism (APE) after cesarean section. A cesarean section was performed on a 27-year old woman with normal course. However, one day after operation, she suddenly developed syncope and dyspnea. Soon after the symptom, she developed hypotension 60 mmHg. As a result of various examinations, her illness was diagnosed as APE with right ventricular dysfunction after cesarean section. She was consulted to our hospital for treatment. Soon after her arrival, we treated her for both APE and cardiogenic shock. The combined with antithrombotic therapy using heparin sodium, was successfully treated the patient from cardiogenic shock due to APE with right ventricular dysfunction after cesarean section.

Effects of steroids on femoral diaphyseal intramedullary circulation in rabbits.

We examined the effect of steroid treatment on the intraosseous femoral vasculature in rabbits, with particular emphasis on the morphologic changes in bone marrow sinusoids and central veins. Adult rabbits were divided into three treatment groups: group A, untreated control; group B, animals were treated with intramuscular injections of methylprednisolone acetate (4 mg/kg/week) for 4 weeks; and group C, rabbits which received injections for 8 weeks. Examination of the intraosseous femoral vasculature was performed using microangiography, scanning electron micrography using a corrosion cast method, and retrograde venography. Bone marrow sinusoids of group C were dilated and the network of anastomosed vessels was irregular compared with those of group A. Changes in group B were intermediate between the control rabbits and group C. Retrograde venography of the central vein showed an irregular course and frequent narrowing of the signal. There were no arterial changes in the steroid-treated groups. These findings indicate that steroid therapy does not induce arterial ischemia but rather intraosseous venous stasis extending from bone marrow sinusoids to the central veins. These changes are attributable to bulging or dilatation of sinusoids, a rise in intraosseous pressure, and reduced blood flow.

Comparison of a new system (Compactdry SCD) with conventional methods for quantitative urine cultures.

AIMS: Compactdry SCD, a new quantitative, ready-to-use and self-diffusible dry medium sheet urine culture system, was compared with conventional methods to evaluate the results of quantitative urine cultures. METHODS & RESULTS: Compactdry SCD was tested on 25 urine specimens, and results compared with those of traditional culture methods. The results from Compactdry SCD analysis correlated well with those from the standard plate count (SPC) method. In fact, the correlation was stronger than that dipslide systems and SPC. Even low-count bacteriuria (< 103 cfu ml(-1) and mixed bacteriuria were detected by Compactdry SCD. CONCLUSIONS: The Compactdry SCD system provides results comparable to those obtained by SPC: simple interpretation, ease of use, long-term storage and good sensitivity. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report suggesting that the Compactdry SCD system has many advantages over traditional quantitative urine culture methods and that it is both appropriate and practical for clinical use.

A phylogenetic-tree analysis elucidating nosocomial transmission of hepatitis C virus in a haemodialysis unit.

Nosocomial transmission of hepatitis C virus (HCV) subtype 1b involving 11 haemodialysis patients occurred in a haemodialysis unit in Japan in March 2000. Sequencing of the HCV-E1 region (411-bp) and phylogenetic-tree analysis showed near identity between HCV isolates derived from these patients and a haemodialysis patient who was known to be HCV-positive. The mode of transmission could not be conclusively established, but retrospective analysis suggested that the sharing of contaminated multidose vials of heparin-saline solutions, which were prepared in the Haemodialysis Center using accidentally contaminated instruments such as needles, may have been responsible for the outbreak. To prevent transmission of HCV in a haemodialysis unit, it may be important to observe strictly standard precautions and to prepare all medications in the Pharmacy. After these measures were taken, no new seroconversions and no new nosocomial transmissions of HCV have been observed in our haemodialysis unit.

Activation of caspases in intestinal villus epithelial cells of normal and nematode infected rats.

BACKGROUND: Small intestinal epithelial cells (IEC) show apoptosis in physiological turnover of cells and in certain inflammatory diseases. AIMS: To investigate the role of caspases in the progression of IEC apoptosis in vivo. METHODS: IEC were separated along the villus-crypt axis from the jejunum of normal and Nippostrongylus brasiliensis infected rats at 4 degrees C. Caspases were examined by a fluorometric assay method, histochemistry, and immunoblotting. RESULTS: Villus cell rich IEC from normal rats exhibited a high level of caspase-3-like activity whereas activities of caspase-1, -8, and -9 were negligible. Immunoblotting analysis of villus cell rich IEC revealed partial cleavage of procaspase-3 into a 17 kDa molecule as well as cleavage of a caspase-3 substrate, poly(ADP-ribose) polymerase (PARP), whereas in crypt cell rich IEC, caspase-3 cleavage was less significant. Caspase-3 activity was also observed histochemically in villus epithelium on frozen sections of the normal small intestine. IEC prepared at 4 degrees C did not reveal nuclear degradation whereas subsequent incubation in a suspension at 37 degrees C induced intense nuclear degradation within one hour in accordance with increases in active caspase-3. This apoptosis was partially suppressed by the caspase inhibitor Z-VAD-fmk. Nematode infected animals showed villus atrophy together with significant increases in levels of caspase-3 in IEC but not of caspase-1, -8, or -9. CONCLUSION: Caspase-3 may have an important role in the physiological replacement of IEC as well as in progression of IEC apoptosis induced by nematode infection.

New anti-malarial flavonol glycoside from Hydrangeae Dulcis Folium.

Bioassay-guided fractionation of the MeOH extract of Hydrangeae Dulcis Folium resulted in isolation of a new flavonol glycoside and two known congeners as anti-malarial principles. These flavonol glycosides showed characteristic proliferation inhibition of Plasmodium falciparum at significantly low concentration without showing any cytotoxicity. In addition, several naturally occurring flavonol glycosides were also shown to exert similar anti-malarial behavior.

Left ventriculoplasty for ischemic cardiomyopathy.

OBJECTIVE: In order to treat ischemic cardiomyopathy, which is defined as non-aneurysmal diffuse akinetic left ventricle with chronic heart failure following myocardial infarction, the mid-term effect of the endoventricular circular patch plasty (EVCPP) was studied. MATERIALS AND METHODS: EVCPP has been performed on 54 patients (46 men and eight women with a mean age of 61 years) during 4 years from March 1997 to December 2000. Thirty-two patients (59%) were NYHA class III and 22 patients (41%) were class IV. Nine patients (17%) had mild angina pectoris before the operation but others had no chest pain. Single, double, triple, and left main disease were noted in six, 13, 32, and three patients, respectively. Mean left ventricular ejection fraction was 23.3 +/- 6.3% (6--30%). Coronary artery bypass grafting was concomitantly undergone by 51 patients (94%) and mitral valve reconstruction was done on 19 patients (35%). RESULTS: Two patients (3.7%) needed an intra-aortic balloon pump to wean from cardiopulmonary bypass. Seven patients (12.9%) died in the hospital. Among them, two patients (4.4%) out of 45 patients who underwent elective operation died of stroke and heart failure. Five patients (55.5%) out of nine patients who required emergency operation died of heart failure and multiorgan failure. Late death occurred in six patients (11.1%) due to arrhythmia and heart failure in each of three patients. Out of 41 survivors, 38 patients returned to NYHA class I or II and three patients to class III. Out of 50 patients who underwent left ventricular study before and after operation, ejection fraction increased from 22.8 +/- 6.6 to 36.2 +/- 8.0% and mean left ventricular end-diastolic volume and left ventricular end-systolic volume indices reduced from 152.8 +/- 24.6 to 105.0 +/- 36.5 and from 113.6 +/- 45.7 to 66.4 +/- 28.4 ml/m(2), respectively. Mean pulmonary wedge pressure decreased from 19.1 +/- 8.8 to 14.9 +/- 6.8 mmHg. One-, 2-, and 3-year actuarial survival rates were 87.9, 82.7 and 77.2%, respectively. CONCLUSION: Left ventriculoplasty using EVCPP is effective to exclude the akinetic LV segment, and left ventricular function and clinical status improve in patients with ischemic cardiomyopathy.

Genetic relationship among Japanese sentinel crabs (Decapoda: Ocypodidae: genus Macrophthalmus).

Seven species (eight populations) of sentinel crabs (genus Macrophthalmus) from the Japan coast and Uca vocans and Ocypode ceratophthalma, were examined electrophoretically for genetic variations in 13 enzymatic and one non-enzymatic protein comprising 17 loci. Most species were highly differentiated from each other (Nei's genetic distance, 0.29-1.63). The least genetic distance was found between M. japonicus and M. banzai, the genetic distinctiveness of the two taxa being supported by three divergent loci with no common allele. The genetic relationships among Macrophthalmus species differed greatly from those inferred from morphological features, with a UPGMA tree suggesting that the sub-genus Macrophthalmus is polyphyletic.