Search

Altered DNA methylation has been previously identified in the spermatozoa of infertile men; however, the origins of these errors are poorly understood. DNA methylation is an epigenetic modification which is thought to play a fundamental role in male germline development. DNA methylation reactions rely on the cellular availability of methyl donors, which are primarily products of folate metabolism, where a key enzyme is methylenetetrahydrofolate reductase ( MTHFR). The MTHFR C677T single nucleotide polymorphism ( SNP) reduces enzyme activity and may potentially alter DNA methylation processes during germline development. The objective of this study was to determine whether altered DNA methylation in spermatozoa is associated with the MTHFR C677T SNP. DNA methylation was evaluated at the H19, IG- GTL2, and MEST imprinted differentially methylated regions in the spermatozoa of 53 men - 44 oligozoospermic men and nine fertile men with normal sperm parameters via bisulfite sequencing of sperm clones. The 44 infertile men were stratified by severity of oligozoospermia - three normal (>15 million spermatozoa/mL), eight moderate (5-15 million spermatozoa/mL), 23 severe (1-5 million spermatozoa/mL), and 10 very severe (<1 million spermatozoa/mL). MTHFR C677T SNP genotyping was conducted in a subset of 44 peripheral blood samples via restriction fragment length polymorphism. A total of three men - severe oligozoospermic and CT genotype - were found to be altered, which is defined as having ≥50% of their clones altered, where an altered clone was in turn defined as ≥50% of CpGs with incorrect DNA methylation patterns. The incidence of three altered men within the CT subgroup, however, was not significantly higher than the incidence in the CC subgroup. Taken together, altered DNA methylation in spermatozoa was not significantly associated with the MTHFR C677T SNP; however, there was a trend for higher incidence of alterations among severe oligozoospermic infertile men with CT genotypes. [ABSTRACT FROM AUTHOR]

Objectives: To describe temporal trends in the use of evidence-based medical therapies and management of patients with acute coronary syndromes (ACS) in Australia and New Zealand.Design, Setting and Participants: Our analysis of the Australian and New Zealand cohort of the Global Registry of Acute Coronary Events (GRACE) included patients with ST-segment-elevation myocardial infarction (STEMI) and non-ST-segment-elevation ACS (NSTEACS) enrolled continuously between January 2000 and December 2007 from 11 metropolitan and rural centres in Australia and New Zealand.Results: 5615 patients were included in this analysis (1723 with STEMI; 3892 with NSTEACS). During 2000-2007 there was an increase in the use of statin therapy, angiotensin-converting enzyme inhibitors or angiotensin receptor blockers, and thienopyridines (P < 0.0001 for each). Among patients with STEMI, there was an increase in emergency revascularisation with PCI (from 11% to 27% [P < 0.0001]), and inhospital coronary angiography (from 61% to 76% [P < 0.0001]). Among patients with NSTEACS, there was an increase in revascularisation with PCI (from 20% to 25% [P = 0.004]). Heart failure rates declined substantially among STEMI and NSTEACS patients (from 21% to 12% [P = 0.0002], and from 13% to 4% [P < 0.0001], respectively) as did rates of hospital readmission for ischaemic heart disease at 6 months (from 23% to 9% [P = 0.0001], and from 24% to 15% [P = 0.0001], respectively).Conclusions: From 2000 to 2007 in Australia and New Zealand, there was a fall in inhospital events and 6-month readmissions among patients admitted with ACS. This showed an association with improved uptake of guideline-recommended medical and interventional therapies. These data suggest an overall improvement in the quality of care offered to contemporary ACS patients in Australia and New Zealand. [ABSTRACT FROM AUTHOR]

The article focuses on the use of cavity ring-down spectroscopy (CRDS) to obtain accurate measurements of carbon dioxide (CO2) and methane (CH4) greenhouse gases in the phase B campaign of Balanço Atmosférico Regional de Carbono na Amazônia in May 2009 in Brazil. It says that measuring the concentration of carbon dioxide is important in studying the carbon cycle and its role in global warming. Results show that CRDS offer more than 0.05 ppm accuracy.

We analyze the North American budget for carbon monoxide using data for CO and formaldehyde concentrations from tall towers and aircraft in a model-data assimilation framework. The Stochastic Time-Inverted Lagrangian Transport model for CO (STILT-CO) determines local to regional-scale CO contributions associated with production from fossil fuel combustion, biomass burning, and oxidation of volatile organic compounds (VOCs) using an ensemble of Lagrangian particles driven by high resolution assimilated meteorology. In many cases, the model demonstrates high fidelity simulations of hourly surface data from tall towers and point measurements from aircraft, with somewhat less satisfactory performance in coastal regions and when CO from large biomass fires in Alaska and the Yukon Territory influence the continental US. Inversions of STILT-CO simulations for CO and formaldehyde show that current inventories of CO emissions from fossil fuel combustion are significantly too high, by almost a factor of three in summer and a factor two in early spring, consistent with recent analyses of data from the INTEXA aircraft program. Formaldehyde data help to show that sources of CO from oxidation of CH4 and other VOCs represent the dominant sources of CO over North America in summer. [ABSTRACT FROM AUTHOR]

Dysregulation of iron homeostasis is implicated in Alzheimer's disease (AD). In this pilot study, common variants of the apolipoprotein E (APOE) and HFE genes resulting in the iron overload disorder of hereditary hemochromatosis (C282Y, H63D and S65C) were evaluated as factors in sporadic AD in an Ontario sample in which folic acid fortification has been mandatory since 1998. Laboratory studies also were done to search for genetic effects on blood markers of iron status, red cell folates and serum B12. Participants included 58 healthy volunteers (25 males, 33 females) and 54 patients with probable AD (20 males, 34 females). Statistical analyses were interpreted at the 95% confidence level. Contingency table and odds ratio analyses supported the hypothesis that in females of the given age range, E4 significantly predisposed to AD in the presence but not absence of H63D. In males, E4 significantly predisposed to AD in the absence of H63D, and H63D in the absence of E4 appeared protective against AD. Among E4+ AD patients, H63D was associated with significant lowering of red cell folate concentration, possibly as the result of excessive oxidative stress. However, folate levels in the lowest population quartile did not affect the risk of AD. A model is presented to explain the experimental findings. [ABSTRACT FROM AUTHOR]

We analyze the North American budget for carbon monoxide using data for CO and formaldehyde concentrations from tall towers and aircraft in a model-data assimilation framework. The Stochastic Time-Inverted, Lagrangian Transport model for CO (STILT-CO) determines local to regional-scale CO contributions associated with production from fossil fuel combustion, biomass burning, and oxidation of volatile organic compounds (VOCs) using an ensemble of Lagrangian particles driven by high resolution assimilated meteorology. In most cases, the model demonstrates high fidelity simulations of hourly surface data from tall towers and point measurements from aircraft, with somewhat less satisfactory performance in coastal regions and when CO from large biomass fires in Alaska and the Yukon Territory influence the continental US. Inversions of STILT-CO simulations for CO and formaldehyde show that current inventories of CO emissions from fossil fuel combustion are significantly too high, by almost a factor of three in summer and a factor two in early spring, consistent with recent analyses of data from the INTEX-A aircraft program. Formaldehyde data help to show that sources of CO from oxidation of CH4 and other VOCs represent the dominant sources of CO over North America in summer. [ABSTRACT FROM AUTHOR]

Although technology holds promising benefits to support older adults at home, attitudes can be a hindrance to the acceptance and use of new technologies. The aim of this study was to explore attitudes of older adults with depressive symptoms using videotelephones (VTs) in their homes. A descriptive design involving 4 participants who received problem solving interventions from a clinical psychologist via VTs was used. Data analysis revealed that the participants' preattitudes were dependent on their active or passive role in the learning process of the new technology. Their postattitudes were classified as ambivalent and positive. Two participants who had a positive attitude toward the VT expressed a positive behavior use. [ABSTRACT FROM AUTHOR]

Observations of CO2 were made in the upper troposphere and lower stratosphere in the deep tropics in order to determine the patterns of large-scale vertical transport and age of air in the Tropical Tropopause Layer (TTL). Flights aboard the NASA WB-57F aircraft over Central America and adjacent ocean areas took place in January and February, 2004 (Pre-AURA Validation Experiment, Pre-AVE) and 2006 (Costa Rice AVE, CR-AVE), and for the same flight dates of 2006, aboard the Proteus aircraft from the surface to 15 km over Darwin, Australia (Tropical Warm Pool International Cloud Experiment, TWP-ICE). The data demonstrate that the TTL is composed of two layers with distinctive features: (1) the lower TTL, 350-360K (potential temperature(θ); approximately 12-14 km), is subject to inputs of convective outflows, as indicated by layers of variable CO2 concentrations, with air parcels of zero age distributed throughout the layer; (2) the upper TTL, from θ=~360K to ~390K (14-18 km), ascends slowly and ages uniformly, as shown by a linear decline in CO2 mixing ratio tightly correlated with altitude, associated with increasing age. This division is confirmed by ensemble trajectory analysis. The CO2 concentration at the level of 360K was 380.0(±0.2) ppmv, indistinguishable from surface site values in the Intertropical Convergence Zone (ITCZ) for the flight dates. Values declined with altitude to 379.2(±0.2) ppmv at 390 K, implying that air in the upper TTL monotonically ages while ascending. In combination with the winter slope of the CO2 seasonal cycle (+10.8±0.4 ppmv/yr), the vertical gradient of -0.78 (±0.09) ppmv gives a mean age of 26(±3) days for the air at 390K and a mean ascent rate of 1.5(±0.3) mms-1. The TTL near 360K in the Southern Hemisphere over Australia is very close in CO2 composition to the TTL in the Northern Hemisphere over Costa Rica, with strong contrasts emerging at lower altitudes (<360 K). Both Pre-AVE and CR-AVE CO2 observed unexpected input from deep convection over Amazônia deep into the TTL. The CO2 data confirm the operation of a highly accurate tracer clock in the TTL that provides a direct measure of the ascent rate of the TTL and of the age of air entering the stratosphere. [ABSTRACT FROM AUTHOR]

The spatial and temporal gradients in atmospheric CO2 contain signatures of carbon fluxes, and as part of inverse studies, these signatures have been combined with atmospheric models to infer carbon sources and sinks. However, such studies have yet to yield finer-scale, regional fluxes over the continent that can be linked to ecosystem processes and ground-based observations. The reasons for this gap are twofold: lack of atmospheric observations over the continent and model deficiencies in interpreting such observations. This paper describes a series of intensive atmospheric sampling field programmes designed as pilot experiments to bridge the observational gap over the continent and to help test and develop models to interpret these observations. We summarize recent results emerging from this work, outlining the role of the intensive atmospheric programmes in collecting CO2 data in both the vertical and horizontal dimensions. These data: (1) quantitatively establish the spatial variability of CO2 and the associated errors from neglecting this variability in models; (2) directly measure regional carbon fluxes from airmass-following experiments and (3) challenge models to reduce and account for uncertainties in atmospheric transport. We conclude with a look towards the future, outlining ways in which intensive atmospheric sampling can contribute towards advancing carbon science. [ABSTRACT FROM AUTHOR]

Small cell carcinoma of the esophagus is a rare and aggressive malignant tumor. Telomerase activation is common in human cancers. There is a lack of data on telomerase activity in esophageal small cell cancers. The present report studied the role of telomerase activity in esophageal small cell carcinoma. The clinicopathologic data of five patients with small cell carcinoma of the esophagus who underwent primary surgical treatment between 1991 and 2000 were studied. Telomeric repeat amplification protocol assays were used to investigate telomerase activity in these tumors. The proliferative activity (MIB-1) and p53 expression of these tumors were also studied using immunohistochemistry and correlated with the telomerase activity. All five small cell carcinomas showed detectable telomerase activity in the primary tumor. Two out of the five morphologically normal esophageal mucosae adjacent to the primary tumor had detectable telomerase activity. There was no correlation between the p53 expression, tumor stage, survival of patients, and the presence of telomerase activity. High MIB-1 expression in esophageal small cell carcinomas was associated with high telomerase activity. Telomerase activation is common in small cell carcinoma of the esophagus. This fact may find application in anti-telomerase treatment for this aggressive tumor. [ABSTRACT FROM AUTHOR]

This study examined the prevalence of human papillomavirus (HPV) infection in a cohort of female sex workers in Singapore. HPV infection was diagnosed by clinical examination, Papanicolaou (Pap) smears, histology and polymerase chain reaction (PCR). One hundred and eighty-seven female sex workers who came for their routine medical screening were enrolled into the study. PCR was positive for HPV in 27 (14.4%), 20 of these had a single HPV type detected, while 7 had multiple types of HPV detected. The most common HPV types identified were types 16, 58 and 18. In conclusion, HPV infection is moderately prevalent among sex workers as tested by PCR, but routine screening with Pap smears does not appear to be a sensitive method of diagnosis for HPV infection. [ABSTRACT FROM AUTHOR]

Metallothioneins (MTs), a group of ubiquitous metalloproteins, comprise isoforms encoded by ten functional genes in humans. Different MT isoforms possibly play different functional roles during development or under various physiological conditions. The MT-1E isoform mRNA has been recently shown to be differentially expressed in oestrogen receptor (OR)-positive and OR-negative breast cancer cell lines. In this study, we evaluated MT-1E mRNA expression via semi-quantitative RT-PCR in 51 primary invasive ductal breast cancer tissues, concurrently with OR-positive and progesterone receptor (PR)-positive MCF7 cells, OR-negative and PR-negative MDA-MB-231 cells and PR-transfected MDA-MB-231 breast cancer cells (ABC28). We demonstrated significantly higher MT-1E mRNA expression in OR-negative compared with OR-positive breast cancer tissues (P = 0.026). MCF7 cells lacked MT-1E mRNA expression, while both OR- and PR-negative MDA-MD-231 cells exhibited a high level of MT-1E mRNA expression. The level of MT-1E mRNA expression in progesterone-treated and -untreated ABC28 cells remained similar as the parental cell line MDA-MB-231-C2 cells. The results suggest that MT-1E may have specific and functional roles in OR-negative invasive ductal breast cancers, possibly mediated via effector genes downstream of the oestrogen receptor, but not through the PR pathway. [ABSTRACT FROM AUTHOR]

A pair of primers were designed for the polymerase chain reaction (PCR) to amplify a 341-base pair fragment of the gene encoding the outer membrane protein IB (PIB) of Neisseria gonorrhoeae. This PCR technique is specific and sensitive, being able to detect gonococcal strains belonging to ten different PIB serovars, but not PIA gonococcus nor other negative control bacteria. PCR products of four representative PIB strains were directly sequenced. Of the three strains belonging to serovar IB4, two (S11 and S48) shared identical nucleotide and amino acid sequences in the PIB region examined. The third IB4 strain (S4) revealed sequences identical to the published IB26 strain (P9). The sequences of strains P9, S4, S11 and S48 were found to differ from those of strain S34 (serovar IB5). The PCR sequencing technique can further differentiate strains belonging to a common serovar and establish clonal relationships among strains. As a molecular epidemiological tool, the PCR-sequencing strategy can augment existing typing methods including serotyping. [ABSTRACT FROM AUTHOR]

A recent epidemic of acute conjunctivitis in Singapore showed again the importance of Coxsackie virus type A24 variant as a causative agent of acute haemorrhagic conjunctivitis (AHC). Although the ocular manifestations appeared similar to those described for the 1970 and 1975 outbreaks, a markedly higher rate of respiratory involvements was noted. Not observed in previous epidemics were herpes-like vesicles in the conjunctiva and eyelids of one patient and vesicles in the buccal mucosa and lips of another from whom Coxsackie virus A24 was isolated. The most interesting finding in this study was the isolation of five wild (non-Sabin) poliovirus type 1 strains. Three strains were obtained from conjunctival and two from throat swabs of patients with mild to severe conjunctivitis. It is conceivable that the rare reports of polio-like paralysis or radiculomyelitis accompanying or following AHC in a few Asian countries could be attributed to concurrent infections with a poliovirus and either enterovirus type 70 or Coxsackie virus type A24. [ABSTRACT FROM PUBLISHER]

This paper describes a microbiological study of 84 young adult men with clinical otitic infections. Gram-negative aerobic bacilli were frequently isolated from these patients diagnosed as having otitis externa or chronic suppurative otitis media, of which Pseudomonasspecies predominated.Staphylococcus aureus, S. epidermidis and aerobic Corynebacterium species (diphtheroids) were also found. About 40% of ear infections were attributed to otomycoses, chiefly from Aspergillus species and Candida parapsilosis. Antimicrobial susceptibility testing of the bacterial isolates revealed that Pseudomonas species were generally resistant to antibiotics commonly employed in general practice: ampicillin, erythromycin, co-trimoxazole, tetracycline and cephaloridine. However, polymjwin B, gentamicin and neomycin were active against some Pseudomonas isolates. Other Gram-negative bacilli were also mainly sensitiveto gentamicin, neomycin as well as co-trimoxazole. Disc diffusion and minimum inhibitor concentration studies demonstrated good activity of ceftazidime, cefoperazone, tobramycin and carbenicillin against strains of Pseudomonas species and other Gram-negative rods. Cefotaxime and cefoxitin were active against Gram-negative bacilli other than Pseudomonas species. Beta-lactamase production did not appear to be the main mechanism of resistance in these communityacquired Gram-negative bacillary isolates. The antimicrobial therapy of otological infections is reviewed. [ABSTRACT FROM PUBLISHER]

AIM: To apply an automated system of nucleic acid hybridisation coupled with the enzyme linked immunosorbent assay (ELISA) for the type specific detection of amplification products of dengue viruses. METHODS: Non-structural 3 (NS3) gene targets of reference strains of all four dengue and other flaviviruses, as well as dengue patient viraemic sera, were subjected to reverse transcription and polymerase chain reaction using consensus and dengue type specific primers and digoxigenin-11-dUTP label incorporation. The amplification products were detected by biotinylated type specific primers which served as ELISA capture probes bound to streptavidin coated tubes. RESULTS: Significantly high spectrophotometric absorbance readings were obtained by hybridisation of the consensus and seminested amplification products of all four dengue viruses with their respective capture probes. In contrast, extremely low absorbances were observed for consensus products of Japanese encephalitis, yellow fever, and Kunjin viruses, which served as negative controls. These ELISA data correlated well with agarose gel electrophoresis of dengue type specific amplified products of diagnostic sizes. CONCLUSIONS: The combination of in vitro amplification and antibody based detection offers rapid, type specific, high throughput, and gel-free detection of amplified products of dengue viruses. [ABSTRACT FROM PUBLISHER]

Consensus primers for the polymerase chain reaction were designed based on conserved motifs within the serine protease and RNA helicase domains encoded by the NS 3 genes of dengue and other flaviviruses. Target fragments of 470 bp were amplified on cDNA templates synthesized from RNAs of dengue types 1, 2, 3, and 4, Japanese encephalitis, Kunjin, and yellow fever viruses using random or specific downstream primers. PCR of oligo(dT)-primed cDNAs from Japanese encephalitis and Kunjin viral RNAs did not yield target bands. As few as 10 copies of dengue viral RNA could be detected. Direct DNA sequencing of PCR products of reference strains of dengue 2 (NGC), Kunjin (MRM 61C) and yellow fever (17 D) viruses demonstrated complete concurrence with published data. However, 2 nucleotide differences were observed between our data for dengue 3 H 87 strain and the published sequence, resulting in a single amino acid disparity. Differences at 21, 16, and 11 nucleotide positions were noted between dengue 1 Hawaii and S 275/90; dengue 4 H 241 and 814669; Japanese encephalitis Nakayama and JaOArS982 viral strains, culminating in only 4, 1 and 1 amino acid residue differences, respectively. These amino acid disparities occurred outside putative active sites of the enzymatic domains, emphasizing the important role of the NS 3 protein in flaviviral replication. This RNA-PCR consensus primer strategy coupled with DNA sequencing represents a valuable tool for the molecular diagnosis and epidemiology of dengue and other flaviviral infections. [ABSTRACT FROM AUTHOR]

We describe a patient who developed a paradoxical embolus to the brain during infusion of bone marrow. She had a patent foramen ovale through which right-to-left shunting led to multiple cerebral emboli. This complication can be prevented by positioning the tip of the infusion catheter in the main pulmonary artery and reducing the volume of marrow product infused. [ABSTRACT FROM AUTHOR]