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.

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.

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.

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.

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.

Increased frequency of autoantibodies in men with sperm antibodies.

Autoantibodies to thyroid microsomes were more frequent in 102 infertile men with complement-dependent sperm-immobilizing activity (sperm immobilization test [SIT] ) in serum (11.8%) than in a control group of 277 men of comparable ages and semen quality without sperm antibodies (4.3%, P less than 0.05). Frequencies of organ-specific antibodies (antigastric parietal cell, antithyroglobulin, and antithyroid microsome) in 57 men with genital tract obstructions and positive SIT were similar to those for control subjects, and there were no significant differences in the frequencies of non-organ-specific autoantibodies (antinuclear antibody, rheumatoid factor, antimitochondrial, and anti-smooth muscle) in the three groups. Because in men without genital tract obstruction antithyroid microsomal autoantibodies were more common with sperm antibodies than without, the possibility of a genetic factor in the causation of sperm autoimmunity should be considered.

Histocompatibility antigens and habitual abortion.

Eighty-five couples with habitual abortion were studied: 43 were primary aborters (i.e., had no advanced pregnancy beyond 20 weeks gestation), and 42 were secondary aborters. The only significant finding in primary female aborters was a reduction in the incidence of B35. In contrast, in secondary female aborters there was a reduction in the incidence of DR3 and A1/B8/DR3 and an increase in the degree of homozygosity of HLA B. In the male partners of primary aborters, there was a reduced incidence of DR3 and A1/B8/DR3. In addition, in male partners of both primary and secondary aborters, there was increased HLA-B homozygosity and increased frequency of DR4-5 phenotype. Increased sharing of HLA antigens between partners was not found to be significantly different from the control population. These findings emphasize that primary and secondary female aborters form distinct populations, which could explain differences in immunological responses to various antigens, including fetal-related antigens. The closely related findings in female secondary aborters and male partners of primary aborters are intriguing but cannot be explained. Whether or not these genetic markers bear any relationship to reproductive success remains to be determined.

The Production of beta 1-specific pregnancy glycoprotein (SP1) by the amnion.

Using indirect immunofluorescence, beta 1-specific pregnancy glycoprotein (SP1) was localised to the amnion cells, as well as to trophoblastic cells. Cultures of fragments of amnion incubated with 3H-leucine for up to 72 h demonstrated that protein was synthesized by the amnion. Using antisera specific to SP1 it was shown that after 72 h incubation about 50% of the total protein could be precipitated. These studies demonstrate that the amnion cells are actively involved in the synthesis of SP1 and could account for the high level of this glycoprotein in amniotic fluid.

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.

The effect of calfhood vaccination with strain 19 on the serological diagnosis and eradication of bovine brucellosis.

Data from 42,224 cattle from 694 herds collected during the brucellosis eradication campaign were used to examine the effects of calfhood strain 19 vaccination. The prevalence to infection in vaccinated herds was 1.8% compared with 9.1% in non-vaccinated herds (p less than 0.005). The mean titre in the former group was lower (p less than 0.001). Vaccinated herds required 3.3 herd tests to achieve a provisionally free status compared with 4.8 in non-vaccinated herds (0.001 less than p less than 0.005). Vaccination did not significantly reduce the number of herd test in herds with less than 100 breeding females. In tests after the initial herd test only 0.5% reactors were found in vaccinated herds compared with 6.9% in non-vaccinated herds (p less than 0.005). There were 0.9% false positive to the Rose Bengal plate test in non-vaccinated and 2.1% in vaccinated animals (p less than 0.005) in non-infected herds. In infected herds this percentage was 3.0% and 4.2% respectively by (p less than 0.05). In the non-infected herds there were 0.04% false positives to the complement fixation tests out of 10,506 non-vaccinated cattle tested and 0.2% out of 24,734 vaccinated cattle.

Effects of topical fluoride treatment on tensile bond strength of pit and fissure sealants.

Clinicians often do not place pit and fissure sealants immediately after topical fluoride treatment because fluoride treatment might reduce bonding to enamel. The effect of NaF, SnF2, and APF on the in-vitro bond strength of two sealants to enamel was determined. The fluoride treatments and control (artificial saliva) were applied to enamel surfaces for four minutes and rinsed. A filled sealant (FluoroShield) and an unfilled sealant (Concise) were bonded to the etched enamel and debonded in tension using a universal testing machine. NaF increased the bond strength of the unfilled sealant; SnF2 and APF had no significant effect. NaF and APF decreased the bond strength of filled sealant while SnF2 had no significant effect. The bond strength of the filled sealant was higher than or equal to that of the unfilled sealant. Exposure of enamel to NaF, SnF2, or APF prior to placement of unfilled or filled sealants has no effect on in vitro bond strength between the enamel and the sealants.

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.

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.

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.

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.