Ellipsometric-based novel DNA biosensor for label-free, real-time detection of Bordetella parapertussis.

Pertussis (or whooping cough) is a contagious disease mainly affecting infants and children and predominantly caused by Bordetella pertussis followed by Bordetella parapertussis. B. parapertussis causes a milder cough but usually symptomatically appears like B. pertussis infection. Thus the epidemiology of illness caused by B. parapertussis is not well understood. In this study, a sensitive and specific method for the rapid diagnosis of B. parapertussis is presented. The covalent immobilization of thiol-terminated DNA oligonucleotides (ss DNA SAM) on a silicon surface by disulfide bond formation is investigated with atomic force microscopy (AFM) and ellipsometry. The measurements indicated an average layer thickness of 5 ± 0.84 nm for 2 µg/µl concentration and 24 h incubation time. This thickness changed to 8.4 ± 0.92 nm for the same concentration (2 µg/µl) by altering the incubation time to 48 h. Ellipsometric data recorded before and after hybridization of B. parapertussis revealed an increase in mean grain area from 91 nm2 to 227 nm2 and a change in the refractive index from 1.489 to 1.648 for 2 µg/µl B. parapertussis, respectively. This change in the refractive index was used to evaluate the amount of adsorbed molecules and their density. The results showed that the density of adsorbed molecules increased from 0.2 to 0.97 g/cm3 after B. parapertussis attachment, respectively. To confirm the hybridization of B. parapertussis to ss DNA SAM, the ds DNA SAM was denatured and the ss DNA SAM surface was reproduced with an average height variation of 6.42 ± 0.75 nm. This showed the stability of the DNA film that can be tuned by varying the concentration and incubation time, thus providing a robust method for the label-free detection of B. parapertussis other than routinely used PCR detection.

Lower gastrointestinal bleeding in pregnancy: Differential diagnosis, assessment and management.

Rectal bleeding is a common symptom experienced by pregnant women. Although the majority of cases are attributable to benign conditions such as haemorrhoids and anal fissures, other more serious diagnoses such as inflammatory bowel disease and malignancy should not be overlooked. Most investigations are safe during pregnancy and these should not be withheld as significant implications on both fetal and maternal morbidity may result. In these cases, a multidisciplinary team approach is essential. This review explores the differential diagnosis, investigation and management of rectal bleeding during pregnancy.

Fiber gene based molecular and biological characterization of hydropericardium-hepatitis syndrome associated avian adenoviruses.

This study was designed to perform biological and molecular characterization of avian adenoviruses (AAVs) recovered from suspected cases of hydropericardium-hepatitis syndrome (HHS) in commercial poultry. Initially the samples were screened by Agar Gel Precipitation Test (AGPT) for the presence of AAVs followed by its confirmation and typing through polymerase chain reaction (PCR) focusing on already reported serotypes AAV-4, AAV-8 and AAV-10 elsewhere. These PCR-positive samples were further subjected to amplification of fiber gene, followed by conducting restriction fragment length polymorphism (RFLP) using restriction enzyme Alu. The selected isolates were further propagated through cell culture and pathogenic potential of selected isolates was determined by infecting chickens. In this study, out of a total 190 samples, 57.8% of suspected cases were found positive for AAV presence through AGPT while sub-type identification using PCR revealed 46.3% for these viruses belonging to AAV-4, 41.8% to AAV-8 and 11.8% showed co-infection of AAV-4 and AAV-8. AAV-10 was not detected in any of the tested samples. On the basis of RFLP pattern, AAV-4 isolates were further divided into four sub-groups (A-D) while AAV-8 isolates had identical RFLP pattern. To further evaluate the pathogenic potential of these sub-groups of AAV-4 isolates, specific pathogen free (SPF) chicks were challenged with selected isolates belonging to each of the sub-groups, resulting in variable pattern of pathogenicity. It is concluded that any variation in the fiber gene of AAV-4 isolates may affect its pathogenicity and eventually specificity of the vaccines used against such infections. Therefore, regular monitoring of the circulating AAV serotypes may be helpful in understanding the pathogenic potential of emerging AAVs, which may lead to development of more effective response strategies accordingly.

Evidence for a paraoxonase-independent inhibition of low-density lipoprotein oxidation by high-density lipoprotein.

One mechanism by which plasma high-density lipoprotein (HDL) may protect against atherogenesis is by inhibiting the oxidation of low-density lipoprotein (LDL). Recent evidence suggests that paraoxonase, an HDL-associated, calcium-dependent enzyme, may be responsible for the antioxidant action of HDL (Mackness et al., Atherosclerosis 1993;104:129; Mackness et al., FEBS Lett 1991;286:152; Watson et al., J Clin Invest 1995;96:2882; Navab et al., Arterio Thromb Vasc Biol 1996;16:831); in particular, paraoxonase activity inhibits the formation of 'minimally oxidized' LDL by hydrolyzing biologically active oxidized phospholipids (Watson et al., J Clin Invest 1995;96:2882; Navab et al., Arterio Thromb Vasc Biol 1996;16:831). However, antioxidant effects of HDL have also been demonstrated under calcium-free conditions, arguing that this enzyme may not be the only mechanism by which HDL inhibits LDL oxidation (Tribble et al., J Lipid Res 1995;36:2580). Here we have evaluated the role of paraoxonase in prevention of LDL oxidation by using HDL subfractions, isolated from human serum or EDTA-plasma, which display markedly different levels of paraoxonase activity; the abilities of modified forms of HDL to prevent LDL oxidation by cultured human (THP-1) macrophages were also assessed. Paraoxonase activity was substantially lower in HDL prepared from plasma compared to serum HDL; moreover, virtually all of the lipoprotein-associated paraoxonase activity was located in the HDL3 fraction, with HDL2 retaining only 1-5% of the total activity. Despite possessing 5-fold differences in paraoxonase activity, HDL3 isolated from plasma or serum was equally effective in inhibiting LDL oxidation by THP-1 macrophages; furthermore, although plasma HDL3 was more protective than plasma HDL2, the latter did significantly inhibit LDL oxidation. Non-paraoxonase antioxidant constituents of plasma HDL3 were investigated further. ApoHDL3, the totally delipidated form of HDL3, was much less effective than native HDL3; when examined individually, purified apolipoprotein A-II gave greater protection than apo A-I, although this effect was not evident in apo A-II-enriched HDL3. Partial delipidation of HDL3, which removes both neutral lipids and alpha-tocopherol, did not significantly diminish its ability to inhibit LDL oxidation by THP-1 macrophages; phospholipid vesicles prepared from partially delipidated HDL3 also inhibited LDL oxidation effectively. We conclude that, in this model of cellular LDL oxidation, the phospholipid fraction of HDL exerts inhibitory effects which are independent of HDL paraoxonase activity.

Acute exacerbation of COPD: factors associated with poor treatment outcome.

OBJECTIVES: To determine the effect of age, severity of lung disease, severity and frequency of exacerbation, steroid use, choice of an antibiotic, and the presence of comorbidity on the outcome of treatment for an acute exacerbation of COPD. DESIGN: A retrospective chart analysis over 24 months. SETTING: A university Veterans Affairs medical center. PATIENTS: Outpatients with COPD who were treated with an antibiotic over a period of 24 months for an acute exacerbation of COPD. METHODS: Severity of an acute exacerbation of COPD was defined using the criteria of Anthonisen et al: increased dyspnea, increased sputum volume, and increased sputum purulence. Severity of lung disease was stratified based on FEV(1) percent predicted using American Thoracic Society guidelines (stage I, FEV(1) > or = 50%; stage II, FEV(1) 35 to 49%; stage III, FEV(1) < 35%). Treatment outcome was judged successful when the patient had no return visit in 4 weeks for a respiratory problem. Failure was defined as a return visit for persistent respiratory symptoms that required a change of an antibiotic in < 4 weeks. RESULTS: One-hundred seven patients with COPD (mean age +/- SD, 66.9 +/- 9.5 years) experienced 232 exacerbations over 24 months. First-line antibiotics (trimethoprim-sulfamethoxazole, ampicillin/amoxicillin, and erythromycin) were used to treat 78% of all exacerbations. Treatment failure was noted in 12.1% of first exacerbations and 14. 7% of all exacerbations, with more than half the failures requiring hospitalization. Host factors that were independently associated with treatment failure included the following: FEV(1) < 35% (46.4% vs 22.4%; p = 0.047), use of home oxygen (60.7% vs 15.6%; p < 0. 0001), frequency of exacerbation (3.8 +/- 2.0 vs 1.6 +/- 0.91; p < 0. 001), history of previous pneumonia (64.3% vs 35.1 p < 0.007), history of sinusitis (28.6% vs 8.8%; p < 0.009) and use of maintenance steroids (32.1% vs 15.2% p = 0.052). Using stepwise logistic regression analysis to identify the top independent variables, the use of home oxygen (p = 0.0002) and frequency of exacerbation (p < 0.0001) correctly classified failures in 83.3% of the patients. Surprisingly, age, the choice of an antibiotic, and the presence of any one or more comorbidity did not affect the treatment outcome. CONCLUSION: The results of our study suggest that patient host factors and not antibiotic choice may determine treatment outcome. Prospective studies in appropriately stratified patients are needed to validate these findings.

Disease and anatomic specificity of ethanolamine plasmalogen deficiency in Alzheimer's disease brain.

A significant and selective deficiency of ethanolamine plasmalogen (PPE) relative to phosphatidylethanolamine was identified in post mortem brain samples from patients with Alzheimer's disease (AD). This lipid defect showed anatomic specificity, being more marked at a site of neurodegeneration in AD brain than in a region relatively spared by the disease (mid-temporal cortex vs. cerebellum) and disease specificity for AD: it was not observed at the primary site of neurodegeneration in Huntington's disease (caudate nucleus) nor Parkinson's disease (substantia nigra). PPE deficiency parallels an inherent tendency towards membrane bilayer instability previously detected in AD brain which is necessarily due to a change in membrane lipid composition, and which may contribute to AD pathogenesis.

Changes in rubisco content in relation to saccharides accumulation in wheat leaves during day.

Two wheat varieties, T. durum (HD 4502) and T. aestivum (Kalyansona) were examined for photosynthesis rate and contents of sugars and rubisco protein in the flag leaf, at forenoon and afternoon at anthesis stage. A decrease in photosynthesis rate was observed in the afternoon compared to forenoon in both the varieties and was associated with an increase in non-reducing sugars and a decrease in rubisco content in the leaves.

A clinical research integration special program (CRISP) for young women with primary ovarian insufficiency.

Large-scale medical sequencing provides a focal point around which to reorganize health care and health care research. Mobile health (mHealth) is also currently undergoing explosive growth and could be another innovation that will change the face of future health care. We are employing primary ovarian insufficiency (POI) as a model rare condition to explore the intersection of these potentials. As both sequencing capabilities and our ability to intepret this information improve, sequencing for medical purposes will play an increasing role in health care beyond basic research: it will help guide the delivery of care to patients. POI is a serious chronic disorder and syndrome characterized by hypergonadotrophic hypogonadism before the age of 40 years and most commonly presents with amenorrhea. It may have adverse health effects that become fully evident years after the initial diagnosis. The condition is most commonly viewed as one of infertility, however, it may also be associated with adverse long-term outcomes related to inadequate bone mineral density, increased risk of cardiovascular disease, adrenal insufficiency, hypothyroidism and, if pregnancy ensues, having a child with Fragile X Syndrome. There may also be adverse outcomes related to increased rates of anxiety and depression. POI is also a rare disease, and accordingly, presents special challenges. Too often advances in research are not effectively integrated into community care at the point of service for those with rare diseases. There is a need to connect community health providers in real time with investigators who have the requisite knowledge and expertise to help manage the rare disease and to conduct ongoing research. Here we review the pathophysiology and management of POI and propose the development of an international Clinical Research Integration Special Program (CRISP) for the condition.

Nucleolar NF-κB/RelA mediates apoptosis by causing cytoplasmic relocalization of nucleophosmin.

In a number of contexts, and particularly in response to cellular stress, stimulation of the NF-kappaB (NF-κB) pathway promotes apoptosis. One mechanism underlying this pro-apoptotic activity is nucleolar sequestration of RelA, which is reported to cause cell death by repressing NF-κB-driven transcription. Here, we identify a novel and distinct nucleolar activity of RelA that induces apoptosis. We demonstrate, using a viral nucleolar localization signal (NoLS)-RelA fusion protein, that direct targeting of RelA to the nucleolus mediates apoptosis, independent of NF-κB transcriptional activity. We demonstrate a requirement for nucleophosmin (NPM, B23.1) in this apoptotic effect, and the apoptotic effect of stress-induced nucleolar RelA. We show by multiple approaches that nucleolar translocation of RelA is causally involved in the relocalization of NPM from the nucleolus to the cytoplasm and that RelA-induced cytoplasmic NPM mediates apoptosis by facilitating the mitochondrial accumulation of BAX. These data uncover a novel stress-response pathway and mechanism by which RelA promotes apoptosis, independent of its effects on NF-κB transcriptional activity. These findings are relevant to the design of novel anticancer agents that target RelA to this compartment.

Clinical trial of an air-abrasion/chemomechanical operative procedure for the restorative treatment of dental patients.

The aim of this study was to investigate whether caries removal with air-abrasion/Carisolv gel is an acceptable and viable alternative in the treatment of dental patients. Twenty-two patients were treated with conventional methods (local anaesthetic injection/drill) followed by alternative treatment (air-abrasion and Carisolv gel) in a general practice setting, by the same operator. The participants' pre-operative anxiety levels were measured using the Modified Dental Anxiety Scale. Their postoperative levels of anxiety/dislike for aspects of both conventional and alternative treatments were assessed using a visual analogue scale. Levels of anxiety/dislike for both treatments were compared and statistically analysed. Results showed 100% of subjects were concerned about several aspects of conventional treatment including pain/discomfort on injection, taste of anaesthetic, length of time tissues remained numb, noise of the drill, its sensory vibrations and water coolant. However, 75% of the study population were happy with all aspects of the air-abrasion technique including dust, pain/discomfort and vibrations produced. Overall, the study population found Carisolv gel to be an acceptable alternative method of caries removal in terms of time taken, pain/discomfort and taste. There were statistically significant differences between patients' perceptions of various aspects of the two treatment methods. All participants found the alternative treatment to be pain-free, quicker and overall more acceptable compared with conventional treatment. The conclusion drawn from the study was that air-abrasion/Carisolv gel treatment was a well-accepted and viable alternative to conventional local anaesthetic injection and drill for dental patients.

Reversal of extrahepatic membrane cholesterol deposition in patients with chronic liver diseases by S-adenosyl-L-methionine.

1. S-Adenosyl-L-methionine is reported to improve serum liver function tests in chronic liver disease. Because liver disease is complicated by cholesterol deposition in hepatic and extrahepatic membranes, we have assessed whether oral administration of S-adenosyl-L-methionine to patients with hepatic disease can reverse the cholesterol enrichment of their erythrocytes. 2. The mean erythrocyte cholesterol-to-phospholipid molar ratio in 13 jaundiced patients was reduced 2 weeks after oral administration of S-adenosyl-L-methionine (from 0.874 +/- 0.112 to 0.844 +/- 0.102, P < 0.05) with 10 of the patients (77%) showing a decrease. By contrast, only four of 11 untreated patients (36%) had a reduced erythrocyte cholesterol-to-phospholipid molar ratio after 2 weeks and the mean values did not differ. 3. The plasma and erythrocyte cholesterol-to-phospholipid molar ratios remained closely correlated (r = 0.77, P < 0.01) before and after treatment, suggesting that S-adenosyl-L-methionine had not acted directly on the cells but rather had improved their lipoprotein milieu. Further support for this concept was provided by following one patient, who initially failed to respond, during an additional 3 weeks of S-adenosyl-L-methionine administration. The plasma cholesterol-to-phospholipid molar ratio fell steadily from week 1 to week 5 and was accompanied by a progressive decrease in the erythrocyte cholesterol-to-phospholipid molar ratio. Moreover, the initially suppressed acetylcholinesterase activity of the erythrocyte membranes returned towards normal during this period. 4. This preliminary study is the first evidence in jaundiced patients that a drug can help to reverse the deposition of cholesterol in an extrahepatic membrane.(ABSTRACT TRUNCATED AT 250 WORDS)