A virome from ornamental flowers in an Australian rural town.

Samples of leaves exhibiting symptoms resembling those caused by virus infection were collected from ornamental street flowers in a rural town in Western Australia. Thirty-seven leaf samples were collected from plants of iris, tulip, lily, daffodil, stock and grape hyacinth. Shotgun sequencing of cDNA derived from leaf samples was done, and analysis showed that about 6% of the sequences obtained were of viral origin. Assembly of virus-like sequences revealed complete or partial genome sequences of 13 virus isolates representing 11 virus species. Eight of the isolates were of potyviruses, one was of a macluravirus, three were of potexviruses, and one was of a bunya-like virus. The complete genome of an isolate originally classified as ornithogalum mosaic virus was genetically divergent and differed in polyprotein cleavage motifs, and we propose that this isolate represents a distinct species. The implications of importing to Australia live plant propagules infected with viruses are discussed.

TBX21 participates in innate immune response by regulating Toll-like receptor 2 expression in Streptococcus pneumoniae infections.

Nasopharyngeal carriage of Streptococcus pneumoniae (pneumococcus) plays an important role in the development of invasive diseases, and is also critically involved in setting up respiratory bacterial and viral infections. We previously reported that pneumococcus, one of the commonly carried bacteria in the nasopharynx, regulates non-typeable Haemophilus influenzae-induced inflammation by upregulating the expression of Toll-like receptor 2 (TLR2). However, the underlying molecular mechanisms by which TLR2 expression is regulated during pneumococcal infections have not yet been well characterized. TBX21 is an important transcription factor of adaptive immunity, but there is an increasing body of evidence pointing to a role in regulating innate immunity. The expression of TBX21 was reported in epithelial cells, but the expression and role of TBX21 in respiratory epithelium, especially for regulating TLR2, has not yet been studied. In this study, we found that pneumococcus upregulates TBX21 expression in the respiratory epithelium. The effect of pneumococcus on TBX21 expression was dependent on its cytoplasmic toxin, pneumolysin. In addition, epithelial TBX21 expression was not regulated by the gram-negative bacterium non-typeable Haemophilus influenzae, peptidoglycan or endotoxin. Deficiency of TBX21 in mice or knocking down TBX21 in epithelial cells suppressed pneumococcus-induced TLR2 expression, but not that of TLR4 or TLR9. These results indicate that the adaptive immune regulator TBX21 participates in regulating innate immune responses, through regulation of TLR2 expression during pneumococcal infections.

Chest CT findings of toxocariasis: correlation with laboratory results.

AIM: To assess the relationship between chest computed tomography (CT) findings of patients with toxocariasis and levels of serological markers. MATERIALS AND METHODS: A total of 38 cases of patients diagnosed with toxocariasis by enzyme-linked immunosorbent assay (ELISA), CT, and serological markers were retrospectively reviewed. The presence of nodule with or without ground-glass opacity (GGO) halo, consolidation, focal GGO, pleural effusion, and lymphadenopathy at chest CT were evaluated. Statistical analysis was performed with the Fisher's exact test. RESULTS: The most common chest CT findings were nodule (n = 12, 31.6%) and focal GGO (n = 12, 31.6%). In patients with normal eosinophil levels, focal GGO (n = 9, 37.5%) was the most common finding. In contrast, nodule with a GGO halo (n = 7, 50%) was the most common finding in the eosinophilia group. Nodule with a GGO halo was more common in the eosinophilia group, with a statistically significant difference (p = 0.017). Nodule was more common in the eosinophilia group, and focal GGO was more common in the normal eosinophil group. CONCLUSION: The most common chest CT findings in toxocariasis were nodule with or without GGO halo, and focal GGO. In the eosinophilia group, nodule with a GGO halo was significantly more frequent. Other CT findings did not show a statistically significant relationship with serological markers.

Resveratrol prevents renal lipotoxicity and inhibits mesangial cell glucotoxicity in a manner dependent on the AMPK-SIRT1-PGC1α axis in db/db mice.

AIMS/HYPOTHESIS: Many of the effects of resveratrol are consistent with the activation of AMP-activated protein kinase (AMPK), silent information regulator T1 (SIRT1) and peroxisome proliferator-activated receptor (PPAR)γ co-activator 1α (PGC-1α), which play key roles in the regulation of lipid and glucose homeostasis, and in the control of oxidative stress. We investigated whether resveratrol has protective effects on the kidney in type 2 diabetes. METHODS: Four groups of male C57BLKS/J db/m and db/db mice were used in this study. Resveratrol was administered via gavage to diabetic and non-diabetic mice, starting at 8 weeks of age, for 12 weeks. RESULTS: The db/db mice treated with resveratrol had decreased albuminuria. Resveratrol ameliorated glomerular matrix expansion and inflammation. Resveratrol also lowered the NEFA and triacylglycerol content of the kidney, and this action was related to increases in the phosphorylation of AMPK and the activation of SIRT1-PGC-1α signalling and of the key downstream effectors, the PPARα-oestrogen-related receptor (ERR)-1α-sterol regulatory element-binding protein 1 (SREBP1). Furthermore, resveratrol decreased the activity of phosphatidylinositol-3 kinase (PI3K)-Akt phosphorylation and class O forkhead box (FOXO)3a phosphorylation, which resulted in a decrease in B cell leukaemia/lymphoma 2 (BCL-2)-associated X protein (BAX) and increases in BCL-2, superoxide dismutase (SOD)1 and SOD2 production. Consequently, resveratrol reversed the increase in renal apoptotic cells and oxidative stress, as reflected by renal 8-hydroxy-deoxyguanosine (8-OH-dG), urinary 8-OH-dG and isoprostane concentrations. Resveratrol prevented high-glucose-induced oxidative stress and apoptosis in cultured mesangial cells through the phosphorylation of AMPK and activation of SIRT1-PGC-1α signalling and the downstream effectors, PPARα-ERR-1α-SREBP1. CONCLUSIONS/INTERPRETATION: The results suggest that resveratrol prevents diabetic nephropathy in db/db mice by the phosphorylation of AMPK and activation of SIRT1-PGC-1α signalling, which appear to prevent lipotoxicity-related apoptosis and oxidative stress in the kidney.

Stem cell transplantation into the intraventricular space via an Ommaya reservoir in a patient with amyotrophic lateral sclerosis.

Stem cells have been highlighted for a possible clinical trial in amyotrophic lateral sclerosis (ALS), and have been employed in SOD1 mice and ALS patients. Human trials for stem cell transplantations have used intrathecal route by lumbar puncture. However, stem cells introduced via lumbar injection would tend to sink downwards rather than ascending to the brain and cervical and thoracic spinal cord. Therefore, we adopted for intraventricular injection via an Ommaya reservoir, and wished to test the practicality of intraventricular stem cell injection in ALS. In the case presented autologous mesenchymal stromal cells (MSCs) were isolated from the bone marrow of a male patient with ALS who underwent insertion of an Ommaya reservoir. Expanded MSCs (hBM-MSCs: dose of 1 X 106 cells/kg) were suspended in autologous CSF and directly transplanted into the ALS patient's lateral ventricle via the Ommaya reservoir. Clinical, laboratory, and radiographic evaluation of the patient revealed no serious adverse effects related to the stem cell therapy. Intraventricular injection with an optimized number of cells is safe, and is a potential route for stem cell therapy in patients with ALS. Intraventricular injection via an Ommaya reservoir makes repetitive injection of stem cells easy and reliable even in far advanced ALS patients.

A novel PRKAR1A mutation in Korean Carney complex family.

Carney complex (CNC) is an autosomal dominant hereditary or sporadic multiple neoplastic syndrome that shows variable clinical symptoms. Generally, CNC appears as skin pigmentation, cardiac or cutaneous myxomas, and multiple endocrine tumours. We performed an extensive evaluation of 9 individuals within 1 family in whom CNC was suspected. Among them, 5 had CNC with various clinical manifestations. We also performed mutational analysis of suspected genes in these patients. Although all patients were members of the same family, variable CNC-related manifestations were observed in each patient. An analysis showed a novel deletion mutation (c.537delA) in exon 6 of the PRKAR1A gene in the patients. Based on our results, the patients were determined to have CNC type I. This is the first such mutational report in Korea.

Microbleeds and free active MMP-9 are independent risk factors for neurological deterioration in acute lacunar stroke.

BACKGROUND AND PURPOSE: in this prospective study, we evaluated mutual relationships amongst microbleeds, matrix metalloproteinase-9 (MMP-9) and neurological deterioration in patients with their first acute lacunar stroke. METHODS: based on diffusion-weighted image findings, we recruited 206 patients with their first acute lacunar stroke. Those without a MRI scan were excluded. Small (a maximum lesion diameter of 15 mm) areas of subcortical gray and white matter with increased signals were considered as lacunar infarctions. GRE images were obtained within 24 h of the onset of stroke symptoms. Venous blood was sampled at base line (within 24 h). Clinical, biochemical, rheological and inflammatory parameters, neurological scales and free, active MMP-9 levels were compared between patients with and without microbleeds. Neurological deterioration was defined as an increase in more than two points on the National Institutes of Health Stroke Scale Score baseline 14 days after the onset of lacunar stroke. RESULTS: of the patients, 79 (38.3%) had microbleeds and 48 (23.3%) showed neurological deterioration. Free, active MMP-9 and C-reactive protein (CRP) levels were significantly increased amongst patients with microbleeds (P < 0.001 and P = 0.047, respectively). Existence of microbleeds (RR = 2.47, 95% CI = 1.25-3.83) and increased free, active MMP-9 (RR = 1.10 per 10 ng/ml, 95% CI = 1.03-1.19) were identified as independent risk factors for neurological deterioration after adjusting for potential confounders. DISCUSSION: ncreased levels of active MMP-9 and the existence of microbleeds might be useful in predicting the deterioration following an initial acute lacunar stroke.

Wide thumb and the first web reconstruction using a neurovascularised instep free flap.

Deformed hands result in palmar defects of the thumb. The sensate instep free flap can be used for wide palmar coverage of the thumb. Three hands with palmar defects of the thumb extended to the first web space underwent soft-tissue reconstruction using a neurovascular instep free flap. These flaps provided sensate coverage with static two-point discrimination values of 8-15 mm. Key pinch strengths of reconstructed thumbs were nearly half of those on the normal side. Donor foot morbidity was minimal with no hyperkeratosis. The neurovascular instep free flap supplies sensate, similar pliable and tough glabrous skin to the palmar surface of the thumb extended to the first web area.

Twin digital and in-step neurovascularised free flaps for reconstruction of the degloved mutilated hands.

BACKGROUND: Degloved mutilated hand injury results in severe contracture of palmar surfaces after avulsion defects of soft tissue and insensate scarring of amputated digits at the proximal interphalangeal level. In an effort to restore the basic function of such hands, we simultaneously used a sensate in-step free flap for re-surfacing the first web space and sensate twin digital free flaps for re-surfacing the palmar defects of the thumb and index finger. METHODS: Three male patients sustained degloved mutilated hand injury from a machine in a factory. The average age of the patients was 26 years. These injuries were reconstructed by concomitant twin digital neurovascularised free flaps harvested on the contralateral hands and the in-step neurovascularised free flaps harvested on the feet. The lateral plantar vascular pedicle of the in-step flaps was anastomosed to the vascular pedicle of the twin digital flaps by a flow-through fashion. RESULT: All flaps survived. These flaps provided durable sensate coverage and improved pinch and grasp. The morbidity of donor fingers and feet was minimal. CONCLUSION: These described flaps supply durable glabrous sensate skin of prehensile function in degloved mutilated hands. Our method is useful in the reconstruction of degloved mutilated hands with amputated stumps of the thumb and counter digit more than 3 cm in length required for pinch and grip.

Serum VEGF levels in acute ischaemic strokes are correlated with long-term prognosis.

BACKGROUND AND PURPOSE: We investigated whether serum vascular endothelial growth factor (VEGF) levels in acute-stage ischaemic stroke patients with small vessel disease (SVD) or large vessel disease (LVD) are correlated with long-term prognoses, based on the difference in NIH Stroke Scale (NIHSS) scores between acute and chronic stages. METHODS: From March 2007 to May 2008, we evaluated patients who experienced an ischaemic stroke for the first time, defined as SVD (n = 89) or LVD (n = 91) using the TOAST classification. Serum samples were taken immediately after admission (within 24 h of stroke onset) to evaluate VEGF levels. After 3 months, follow-up NIHSS scores were collected for all patients. RESULTS: Serum VEGF levels in the acute stage (within 24 h of stroke onset) were higher in the LVD group than in the SVD group and were correlated with infarction volume. The increase in serum VEGF levels in the acute stage was proportional to an improved NIHSS score after 3 months. After adjustment for covariates, serum VEGF levels in the acute stage were still significantly correlated with the long-term prognosis of ischaemic stroke. CONCLUSION: Serum VEGF levels are correlated with long-term prognoses in acute ischaemic stroke patients.

Low rate of total hip replacement as reflected by a low prevalence of hip osteoarthritis in South Korea.

OBJECTIVE: We attempted to estimate the rate of total hip replacement (THR) using a national database and the prevalence of hip osteoarthritis (OA) from the reading of intravenous pyelograms (IVPs) in a Korean population. MATERIALS: Reimbursement records from all hospitals in South Korea were extracted from the Health Insurance Review Agency (HIRA) database. Records with both the procedure code corresponding to THR and containing the diagnosis code for hip OA were selected. We estimated the age- and sex-specific rates of THR from 2002 to 2006. Hip joints from 580 subjects older than 70 years old who underwent an IVP were assessed for the presence of OA. RESULTS: The rate of THR increased with age, reaching a peak over the age of 65-69 years, with the age-standardized risk ratios in women vs men of approximately 1.5. Although the rate of THR increased over the 5-year study period, it was significantly lower than that of total knee replacement (TKR) in Korean population (THR vs TKR 1:15.9). The prevalence of hip OA in the IVP cohort was 1.2% (1.7% for men and 0.7% for women). CONCLUSION: The rate of THR was significantly lower than that of TKR in Korean population. Hip OA prevalence among the IVP subjects was 1.2%. Further studies on factors that account for the low prevalence of hip OA among Asians need to be conducted.

Haemodynamic effects of oxytocin given as i.v. bolus or infusion on women undergoing Caesarean section.

BACKGROUND: The cardiovascular effects of oxytocin in animal models and women undergoing Caesarean section include tachycardia, hypotension and decrease in cardiac output. These can be sufficient to cause significant compromise in high-risk patients. We aimed to find a simple way to decrease these risks whilst retaining the benefits of oxytocin in decreasing bleeding after delivery. Method. We recruited 30 women undergoing elective Caesarean section. They were randomly allocated to receive 5 u of oxytocin either as a bolus injection (bolus group) or an infusion over 5 min (infusion group). These women had their heart rate and intra-arterial blood pressure recorded every 5 s throughout the procedure. The haemodynamic data, along with the estimated blood loss, were compared between the groups. RESULTS: Marked cardiovascular changes occurred in the bolus group; the heart rate increased by 17 (10.7) beats min(-1) [mean (sd)] compared with 10 (9.7) beats min(-1) in the infusion group. The mean arterial pressure decreased by 27 (7.6) mm Hg in the bolus group compared with 8 (8.7) mm Hg in the infusion group. There were no differences in the estimated blood loss between the two groups. CONCLUSION: We recommend that bolus doses should be used with caution, and further studies should ascertain if oxytocin is equally effective in reducing blood loss when given at a slower rate.

Ocular flutter induced only by optokinetic stimulation.

Ocular flutter (OF) refers to occasional bursts of involuntary horizontal oscillation around the point of fixation, characterised by rapid horizontal repetitive oscillation without intersaccadic interval in electro-ocular recording. In spite of numerous reports of OF, there has been no previous report describing OF developed by simply looking at moving objects. We report a patient with olivopontocerebellar atrophy (OPCA), with OF induced only by optokinetic stimulation (OKS). A 65-year-old man, with OPCA complained of dizziness and oscillopsia occurring only with a specific type of eye movement. The electro-oculographic recording showed a decreased gain of pursuit, catch-up saccade, and decreased velocity of saccade, and OF was induced only by OKS of more than 40 degrees /second. In OPCA, partial loss of omnipause neurons and/or abnormal firing of burst neurons, especially those involved in optokinetic movement, could be the cause of OF occurring only with OKS.

Spinocerebellar ataxia type 6 and episodic ataxia type 2 in a Korean family.

Spinocerebellar ataxia type 6 (SCA6), episodic ataxia type 2 (EA2) and familial hemiplegic migraine (FHM) have been known as allelic disorders, which are caused by the alteration of the alpha1A voltage-dependent calcium channel subunit. Expansions of the CAG repeat in the CACNA1A gene on the short arm of the chromosome 19 induce SCA6, and point mutations in the same gene are responsible for EA2 and FHM. In recent studies, both SCA6 and EA2 have been concurrently found in families with 26 CAG repeats without previously reported point mutations either in coding sequences or in intron-exon junctions. We describe a Korean family with CAG26 repeats in the CACNA1A gene. Some of the affected family members had progressive ataxia typical of SCA6 whereas others had episodic vertigo responsive to acetazolamide typical of EA2. Our family support that SCA6 and EA2 are allelic disorders with a high phenotypic variability.

A high-resolution add-on in-lens attachment for scanning electron microscopes.

A compact add-on objective lens for the scanning electron microscope (SEM) has been designed and tested. The lens is < 35 mm high and can be fitted on to the specimen stage as an easy-to-use attachment. Initial results show that it typically improves the spatial resolution of the SEM by a factor of three. The add-on unit is based upon a permanent magnet immersion lens design. Apart from the extra attachment to the specimen stage, the SEM with the add-on lens functions in the normal way. The in-lens unit can comfortably accommodate specimen heights up to 10 mm. The new add-on lens unit opens up the possibility of operating existing SEMs in the high-resolution in-lens mode. By using a deflector at the top of the add-on lens unit, it can also operate as a quantitative multichannel voltage contrast spectrometer, capable of recording the energy spectrum of the emitted secondary electrons. Initial experiments confirm that a significant amount of voltage contrast can be obtained.

Tensile bond strengths of fourth- and fifth-generation dentin adhesives with packable resin composites.

PURPOSE: Placement of packable composites would be simplified if a single-bottle dentin bonding agent, rather than the more complex, multiple-bottle dentin bonding agents, involving multiple steps, could be used successfully with adequate bond strength. This study was designed to compare the tensile bond strengths to extracted human dentin of four single-bottle (fifth-generation) and four multiple-bottle (fourth-generation) dentin bonding agents with four packable composites. MATERIALS AND METHODS: Packable composites (P60, 3M Dental Products Division; ALERT, Jeneric-Pentron; Pyramid Dentin, Bisco; and Clearfil AP-X, Kuraray America) were tested for in vitro tensile bond strength to human dentin with their single-bottle and their multiple-bottle dentin bonding agents. Bond strengths were measured using a truncated cone tensile test, with a bonding diameter of 3 mm, after storage of specimens in water at 37 degrees C and 100% relative humidity for 24 hours, and using a testing machine at a crosshead speed of 0.5 mm per minute. RESULTS: Means and standard deviations (n = 10) of tensile bond strengths (MPa) for the single-bottle system were P60, 21.3 MPa (6.7); ALERT, 26.6 MPa (4.2); Pyramid Dentin, 14.8 MPa (4.4); and Clearfil AP-X, 18.6 MPa (3.7). For the multiple-bottle system they were P60, 20.0 MPa (8.6); ALERT, 20.6 MPa (6.2); Pyramid Dentin, 16.0 MPa (3.4); and Clearfil AP-X, 17.8 MPa (6.0). Two-way analysis of variance showed significant differences among composites but no significant differences between single-bottle and multiple-bottle bonding agents. The Tukey-Kramer interval (p < .05) for comparisons of bond strengths among composites was 4.8 MPa. CONCLUSIONS: For single-bottle agents, bond strengths were ranked as ALERT > P60, P60 = Clearfil, P60 > Pyramid, and Clearfil = Pyramid. For multiple-bottle agents, there were no significant differences in bond strengths among the composites. CLINICAL SIGNIFICANCE: Single-bottle bonding agents provide similar in vitro bond strengths for use with packable composites as do the same manufacturers' multiple-bottle bonding agents.

Identification and characterization of the major allergens of buckwheat.

BACKGROUND: Buckwheat (BW) has been recognized as a common food allergen in Korea, Japan, and other countries. Until now, serologic findings of BW food-allergic patients and its major allergenic components have not been clarified. In this study, we analyzed the serologic findings of BW food allergy and characterized its major allergenic components. METHODS: Nineteen BW-allergic subjects with symptoms after BW ingestion and 15 asymptomatic control subjects with positive skin prick test to BW were recruited. BW-specific IgE was measured with the Pharmacia CAP kit. Allergenic components of BW were analyzed by IgE immunoblotting, periodate oxidation, two-dimensonal PAGE, and sequencing of N-terminal amino acids. RESULTS: From the BW-allergic patients and asymptomatic controls, the sensitivity (100%), specificity (53%), and negative (100%) and positive predictive values (73%) of Pharmacia CAP specific IgE for diagnosis were estimated. The prevalence of IgE binding to 24-kDa (pI 8.3), 16-kDa (pI 5.6), and 9-kDa (pI 5.0/ 6.0) allergens was higher than 50% in BW-allergic and asymptomatic subjects. However, the specific IgE to split 19-kDa (pI 6.5/7.0) allergens were more specifically found in BW-allergic patients than in asymptomatic subjects (78% vs 7%). N-terminal amino-acid sequences of 19-kDa and 16-kDa allergens showed moderate and weak homology to the 19-kDa globulin protein of rice and alpha-amylase/trypsin inhibitor of millet, respectively. The N-terminus of the 9-kDa isoallergens were not different from each other and were identified as the reported trypsin inhibitors of BW. Attenuation of the IgE binding to the 9-kDa allergen was found with periodate oxidation. CONCLUSIONS: The allergens of 24, 19, 16, and 9 kDa are strong candidates to be major allergens, and the 19-kDa allergen was relatively specific for BW-allergic patients. Moreover, measurement of BW-specific IgE and the features of immunoblotting should be very useful tools in the diagnosis of BW allergy.

Changes in expression of sodium cotransporters and aquaporin-2 during ischemia-reperfusion injury in rabbit kidney.

Ischemic renal injury is associated with defects in transport functions of the proximal tubules and urinary concentration ability. To determine whether alterations in expression of various transporter genes contribute to an impairment in renal functions, the expression of various solute transport genes was analyzed in renal cortex and medulla of rabbits with ischemic acute renal failure. Rabbits were subjected to 60 min of renal pedicle clamping followed by 24, 48, or 72 h of reperfusion. Urine volume and glomerular filtration rate were markedly decreased, which were accompanied by an increase in serum creatinine level and fraction Na+ excretion. Glucosuria and phosphaturia were evident during reperfusion periods. These alterations in renal functions were persisted to 72 h after reperfusion. The Na+-dependent uptakes of glucose and phosphate by brush border membrane vesicles were inhibited by 24 h of reperfusion. mRNA levels for Na+-glucose, Na+-phosphate, and Na+-succinate cotransporter analyzed by RT-PCR were not changed by 60 min of ischemia alone, but were significantly reduced by 24 h of reperfusion. mRNA levels for apical Na+-K+-2Cl- cotransporter, NaCl cotransporter, and turea transporter in the medulla were not changed during reperfusion. Protein levels for AQP2 in the medulla, but not AQP1 in the cortex, analyzed by Western blot were significantly reduced at 24 h after reperfusion. These results suggest that reductions in expression of Na+-cotransporter genes in the proximal tubules may be important factors in the impairment in Na+-dependent reabsorption of solutes and that decrease in AQP2 protein may be involved in defect in urinary concentration ability in rabbits with ischemic acute renal failure.