Development of Novel Assays for Measuring Different Molecular Forms of Prostate Specific Antigen

Prostate cancer (PCa) is the most commonly diagnosed non-skin cancer and second leading cause of cancer-related death of men in developed countries. Measurement of prostate specific antigen (PSA) is a very sensitive method for diagnosing and monitoring of prostate cancer (PCa), but the specificity needs improvement. Measurements of different molecular forms of PSA have been shown to improve differentiation between PCa and benign prostatic diseases. However, accurate measurement of some isoforms has not been achieved in previous assays. The aim of the present study was to develop new assays that reliably measure enzymatically active PSA, PSA-α1-chymotryposin (PSA-ACT) and PSA-α1-protease inhibitor (PSA-API), and to evaluate their diagnostic value. Double-label immunofluorometric assays using a novel monoclonal antibody (MAb) and another antibody to either free PSA (fPSA) or total PSA (tPSA) were developed and used to measure PSA-ACT and fPSA or tPSA at the same time. These assays provide enough sensitivity for measurement of PSA-ACT in sera with low PSA levels. The results obtained confirmed that proportion of PSA-ACT to tPSA (%PSA-ACT) was as useful as proportion of fPSA to tPSA (%fPSA) for discrimination between PCa and benign prostatic hyperplasia (BPH). We developed an immunoassay for detection of PSA-API based on proximity ligation, which improved assay sensitivity 10-fold compared with conventional assays. Our results confirmed previous findings that the PSA-API level is somewhat lower in men with than without PCa, and the combination of %fPSA and proportion of PSA-API to tPSA (%PSA-API) provides diagnostic improvement compared with either method alone. Assays based on this principle should be applicable to other immunoassays in which the nonspecific background is a problem. An immunopeptidometric sandwich assay (IPMA) was developed to measure the enzymatically active PSA. This assay showed high specificity, but sensitivity was not good enough for measurement of PSA concentrations in the gray zone, 2-10 µg/L, in which tPSA does not efficiently differentiate between PCa and BPH. We further developed a solid-phase proximity ligation immunoassay, which provided a 10-fold improvement in sensitivity. This proof of concept study shows that peptides reacting with proteins are potentially useful for sensitive and specific measurement of protein variants for which specific MAbs cannot be obtained.

Aqueous colloidal stability evaluated by zeta potential measurement and resultant TiO2 for superior photovoltaic performance

Controlling the morphological structure of titanium dioxide (TiO 2) is crucial for obtaining superior power conversion efficiency for dye-sensitized solar cells. Although the sol-gel-based process has been developed for this purpose, there has been limited success in resisting the aggregation of nanostructured TiO2, which could act as an obstacle for mass production. Herein, we report a simple approach to improve the efficiency of dye-sensitized solar cells (DSSC) by controlling the degree of aggregation and particle surface charge through zeta potential analysis. We found that different aqueous colloidal conditions, i.e., potential of hydrogen (pH), water/titanium alkoxide (titanium isopropoxide) ratio, and surface charge, obviously led to different particle sizes in the range of 10-500 nm. We have also shown that particles prepared under acidic conditions are more effective for DSSC application regarding the modification of surface charges to improve dye loading and electron injection rate properties. Power conversion efficiency of 6.54%, open-circuit voltage of 0.73 V, short-circuit current density of 15.32 mA/cm2, and fill factor of 0.73 were obtained using anatase TiO 2 optimized to 10-20 nm in size, as well as by the use of a compact TiO2 blocking layer.

Sinteractivity, proton conductivity and chemical stability of BaZr 0.7In0.3O3-δ for solid oxide fuel cells (SOFCs)

In3+ was used as dopant for BaZrO3 proton conductor and 30 at%-doped BaZrO3 samples (BaZr0.7In 0.3O3-δ, BZI) were prepared as electrolyte materials for proton-conducting solid oxide fuel cells (SOFCs). The BZI material showed a much improved sinteractivity compared with the conventional Y-doped BaZrO 3. The BZI pellets reached almost full density after sintering at 1600 °C for 10 h, whereas the Y-doped BaZrO3 samples still remained porous under the same sintering conditions. The conductivity measurements indicated that BZI pellets showed smaller bulk but improved grain boundary proton conductivity, when compared with Y-doped BaZrO3 samples. A total proton conductivity of 1.7 × 10-3 S cm -1 was obtained for the BZI sample at 700 °C in wet 10% H 2 atmosphere. The BZI electrolyte material also showed adequate chemical stability against CO2 and H2O, which is promising for application in fuel cells.

Absolute quantification of human tear lactoferrin using multiple reaction monitoring technique with stable-isotopic labeling

The mass spectrometry technique of multiple reaction monitoring (MRM) was used to quantify and compare the expression level of lactoferrin in tear films among control, prostate cancer (CaP), and benign prostate hyperplasia (BPH) groups. Tear samples from 14 men with CaP, 15 men with BPH, and 14 controls were analyzed in the study. Collected tears (2 μl) of each sample were digested with trypsin overnight at 37 °C without any pretreatment, and tear lactoferrin was quantified using a lactoferrin-specific peptide, VPSHAVVAR, both using natural/light and isotopic-labeled/heavy peptides with MRM. The average tear lactoferrin concentration was 1.01 ± 0.07 μg/μl in control samples, 0.96 ± 0.07 μg/μl in the BPH group, and 0.98 ± 0.07 μg/μl in the CaP group. Our study is the first to quantify tear proteins using a total of 43 individual (non-pooled) tear samples and showed that direct digestion of tear samples is suitable for MRM studies. The calculated average lactoferrin concentration in the control group matched that in the published range of human tear lactoferrin concentration measured by enzyme-linked immunosorbent assay (ELISA). Moreover, the lactoferrin was stably expressed across all of the samples, with no significant differences being observed among the control, BPH, and CaP groups.

Influence of spatial variability of permeability property on steady state seepage flow and slope stability analysis

In recent years, spatial variability modeling of soil parameters using random field theory has gained distinct importance in geotechnical analysis. In the present Study, commercially available finite difference numerical code FLAC 5.0 is used for modeling the permeability parameter as spatially correlated log-normally distributed random variable and its influence on the steady state seepage flow and on the slope stability analysis are studied. Considering the case of a 5.0 m high cohesive-frictional soil slope of 30 degrees, a range of coefficients of variation (CoV%) from 60 to 90% in the permeability Values, and taking different values of correlation distance in the range of 0.5-15 m, parametric studies, using Monte Carlo simulations, are performed to study the following three aspects, i.e., (i) effect ostochastic soil permeability on the statistics of seepage flow in comparison to the analytic (Dupuit's) solution available for the uniformly constant permeability property; (ii) strain and deformation pattern, and (iii) stability of the given slope assessed in terms of factor of safety (FS). The results obtained in this study are useful to understand the role of permeability variations in slope stability analysis under different slope conditions and material properties. (C) 2009 Elsevier B.V. All rights reserved.

Analysis of stability of earthen dams in kachchh region, Gujarat, India

The Kachchh region of Gujarat, India bore the brunt of a disastrous earthquake of magnitude M-w=7.6 that occurred on January 26, 2001. The major cause of failure of various structures including earthen dams was noted to be the presence of liquefiable alluvium in the foundation soil. Results of back-analysis of failures of Chang, Tappar, Kaswati and Rudramata earth dams using pseudo-static limit equilibrium approach presented in this paper confirm that the presence of liquefiable layer contributed to lesser factors of safety leading to a base type of failure that was also observed in the field. Following the earthquake, earth dams have been rehabilitated by the concerned authority and it is imperative that the reconstructed sections of earth dams be reanalyzed. It is also increasingly realized that risk assessment of dams in view of the large-scale investment made and probabilistic analysis is necessary. In this study, it is demonstrated that the probabilistic approach when used in conjunction with deterministic approach helps in providing a rational solution for quantification of safety of the dam and in the estimation of risk associated with the dam construction. (C) 2007 Elsevier B.V. All rights reserved.

A CD8(+) T cell clone specific for antigen also recognizes peptidomimics present in anti-idiotypic antibody: Implications for T cell memory

The relay hypothesis [R. Nayak, S. Mitra-Kaushik, M.S. Shaila, Perpetuation of immunological memory: a relay hypothesis, Immunology 102 (2001) 387-395] was earlier proposed to explain perpetuation of immunological memory without requiring long lived memory cells or persisting antigen. This hypothesis envisaged cycles of interaction and proliferation of complementary idiotypic B cells (Burnet cells) and anti-idiotypic B cells (Jerne cells) as the primary reason for perpetuation of immunological memory. The presence of pepti-domimics of antigen in anti-idiotypic antibody and their presentation to antigen specific T cells was postulated to be primary reason for perpetuation of T cell memory. Using a viral hemagglutinin as a model, in this work, we demonstrate the presence of peptidomimics in the variable region of ail anti-idiotypic antibody capable of functionally mimicking the antigen derived peptides. A CD8(+) CTL clone was generated against the hemagglutinin protein which specifically responds to either peptidomimic synthesizing cells or peptidomimic pulsed antigen presenting cells. Thus, it appears reasonable that a population of activated antigen specific T cells is maintained in the body by presentation of peptidomimic through Jerne cells and other antigen presenting cells long after immunization. (C) 2007 Elsevier Inc. All rights reserved.

Effect of residual stresses and metallographic stability on the over all performance of integral diaphragm material

The integral diaphragm pressure transducer consists of a diaphragm machined from precipitation hardened martensitic (APX4) steel. Its performance is quite significant as it depends upon various factors such as mechanical properties including induced residual stress levels, metallurgical and physical parameters due to different stages of processing involved. Hence, the measurement and analysis of residual stress becomes very important from the point of in-service assessment of a component. In the present work, the stress measurements have been done using the X-ray diffraction (XRD) technique, which is a non-destructive test (NDT). This method is more reliable and widely used compared to the other NDT techniques. The metallurgical aspects have been studied by adopting the conventional metallographic practices including examination of microstructure using light microscope. The dimensional measurements have been carried out using dimensional gauge. The results of the present investigation reveals that the diaphragm material after undergoing series of realization processes has yielded good amount of retained austenite in it. Also, the presence of higher compressive stresses induced in the transducer results in non-linearity, zero shift and dimensional instability. The problem of higher retained austenite content and higher compressive stress have been overcome by adopting a new realization process involving machining and cold and hot stabilization soak which has brought down the retained austenite content to about 5–6% and acceptable level of compressive stress in the range −100 to −150 MPa with fine tempered martensitic phase structure and good dimensional stability. The new realization process seems to be quite effective in terms of controlling retained austenite content, residual stress, metallurgical phase as well as dimensional stability and this may result in minimum zero shift of the diaphragm system.

Developing an Unmanned Aerial Vehicle (UAV) multi-sensor payload carrier for air quality monitoring

Emissions of gases and particles from sea-faring ships have been shown to impact on the atmospheric chemistry and climate. To efficiently monitor and report these emissions found from a ship’s plume, the concept of using a multi-rotor or UAV to hover inside or near the exhaust of the ship to actively record the data in real time is being developed. However, for the required sensors obtain the data; their sensors must face into the airflow of the ships plume. This report presents an approach to have sensors able to read in the chemicals and particles emitted from the ship without affecting the flight dynamics of the multi-rotor UAV by building a sealed chamber in which a pump can take in the surrounding air (outside the downwash effect of the multi-rotor) where the sensors are placed and can analyse the gases safely. Results show that the system is small, lightweight and air-sealed and ready for flight test.

Thrombophilia and direct thrombin inhibitor lepirudin clinical and monitoring aspects

Thrombophilia (TF) predisposes both to venous and arterial thrombosis at a young age. TF may also impact the thrombosis or stenosis of hemodialysis (HD) vascular access in patients with end-stage renal disease (ESRD). When involved in severe thrombosis TF may associate with inappropriate response to anticoagulation. Lepirudin, a potent direct thrombin inhibitor (DTI), indicated for heparin-induced thrombocytopenia-related thrombosis, could offer a treatment alternative in TF. Monitoring of narrow-ranged lepirudin demands new insights also in laboratory. The above issues constitute the targets in this thesis. We evaluated the prevalence of TF in patients with ESRD and its impact upon thrombosis- or stenosis-free survival of the vascular access. Altogether 237 ESRD patients were prospectively screened for TF and thrombogenic risk factors prior to HD access surgery in 2002-2004 (mean follow-up of 3.6 years). TF was evident in 43 (18%) of the ESRD patients, more often in males (23 vs. 9%, p=0.009). Known gene mutations of FV Leiden and FII G20210A occurred in 4%. Vascular access sufficiently matured in 226 (95%). The 1-year thrombosis- and stenosis-free access survival was 72%. Female gender (hazards ratio, HR, 2.5; 95% CI 1.6-3.9) and TF (HR 1.9, 95% CI 1.1-3.3) were independent risk factors for the shortened thrombosis- and stenosis-free survival. Additionally, TF or thrombogenic background was found in relatively young patients having severe thrombosis either in hepatic veins (Budd-Chiari syndrome, BCS, one patient) or inoperable critical limb ischemia (CLI, six patients). Lepirudin was evaluated in an off-label setting in the severe thrombosis after inefficacious traditional anticoagulation without other treatment options except severe invasive procedures, such as lower extremity amputation. Lepirudin treatments were repeatedly monitored clinically and with laboratory assessments (e.g. activated partial thromboplastin time, APTT). Our preliminary studies with lepirudin in thrombotic calamities appeared safe, and no bleeds occurred. An effective DTI lepirudin calmed thrombosis as all patients gradually recovered. Only one limb amputation was performed 3 years later during the follow-up (mean 4 years). Furthermore, we aimed to overcome the limitations of APTT and confounding effects of warfarin (INR of 1.5-3.9) and lupus anticoagulant (LA). Lepirudin responses were assessed in vitro by five specific laboratory methods. Ecarin chromogenic assay (ECA) or anti-Factor IIa (anti-FIIa) correlated precisely (r=0.99) with each other and with spiked lepirudin in all plasma pools: normal, warfarin, and LA-containing plasma. In contrast, in the presence of warfarin and LA both APTT and prothrombinase-induced clotting time (PiCT®) were limited by non-linear and imprecise dose responses. As a global coagulation test APTT is useful in parallel to the precise chromogenic methods ECA or Anti-FIIa in challenging clinical situations. Lepirudin treatment requires multidisciplinary approach to ensure appropriate patient selection, interpretation of laboratory monitoring, and treatment safety. TF seemed to be associated with complicated thrombotic events, in venous (BCS), arterial (CLI), and vascular access systems. TF screening should be aimed to patients with repeated access complications or prior unprovoked thromboembolic events. Lepirudin inhibits free and clot-bound thrombin which heparin fails to inhibit. Lepirudin seems to offer a potent and safe option for treatment of severe thrombosis. Multi-centered randomized trials are necessary to assess the possible management of complicated thrombotic events with DTIs like lepirudin and seek prevention options against access complications.

Fuzzy-logic-based health monitoring and residual-life prediction for composite helicopter rotor

A health-monitoring and life-estimation strategy for composite rotor blades is developed in this work. The cross-sectional stiffness reduction obtained by physics-based models is expressed as a function of the life of the structure using a recent phenomenological damage model. This stiffness reduction is further used to study the behavior of measurable system parameters such as blade deflections, loads, and strains of a composite rotor blade in static analysis and forward flight. The simulated measurements are obtained using an aeroelastic analysis of the composite rotor blade based on the finite element in space and time with physics-based damage modes that are then linked to the life consumption of the blade. The model-based measurements are contaminated with noise to simulate real data. Genetic fuzzy systems are developed for global online prediction of physical damage and life consumption using displacement- and force-based measurement deviations between damaged and undamaged conditions. Furthermore, local online prediction of physical damage and life consumption is done using strains measured along the blade length. It is observed that the life consumption in the matrix-cracking zone is about 12-15% and life consumption in debonding/delamination zone is about 45-55% of the total life of the blade. It is also observed that the success rate of the genetic fuzzy systems depends upon the number of measurements, type of measurements and training, and the testing noise level. The genetic fuzzy systems work quite well with noisy data and are recommended for online structural health monitoring of composite helicopter rotor blades.

Determination of the thermodynamic stability of TiB2

The standard free energy of formation of titanium boride (TiB2) Was measured by the Electro Motive Force (EMF) method (by using yttria doped thoria (YDT) as the solid electrolyte). Two galvanic cells viz. Cell (I): Pt, TiB2 (s), TiO2 (s), B (s) vertical bar YDT vertical bar NiO (s), Ni (s), Pt and cell (II): Pt, TiB2 (s), TiO2 (s), B (s) vertical bar YDT vertical bar FeO (s). Fe (s), Pt were constructed in order to determine the Delta(f)G degrees, of TiB2. Enthalpy increments on TiB2 were measured by using inverse drop calorimetry over the temperature range 583-1769 K. The heat capacity, entropy and the free energy function have been derived from these experimental data in the temperature range 298-1800 K. The mean value of the standard enthalpy of formation of TiB2 (Delta H-f(298)degrees (TiB2)) was obtained by combining these Delta(f)G degrees, values and the free energy functions of TiB2 derived from the drop calorimetry data. The mean values of Delta H-f(298)degrees (TiB2) derived from the Delta(f)G degrees, data obtained from cell I and II were -322 +/- 1.2 kJ mol(-1) and -323.3 +/- 2.1 kJ mol(-1), respectively. These values were found to be in very good agreement with the assessed data. (C) 2009 Elsevier B.V. All rights reserved.

Temperature monitoring of nonanaesthetised patients in a cardiac catheterisation laboratory

Aim and objectives To identify the prevalence that temperature reduced by more than 1°C from pre to post-procedure in a sample of non-anaesthetised patients undergoing procedures in a cardiac catheterisation laboratory. Background Advances in medical technology are minimising the invasiveness of diagnostic tests and treatments for disease, which is correspondingly increasing the number of medical procedures performed without sedation or anaesthesia. Procedural areas in which medical procedures are performed without anaesthesia are typically kept at a cool temperature for staff comfort. As such, there is a need to inform nursing practices in regard to the thermal management of non-anaesthetised patients undergoing procedures in surgical or procedural environments. Design Single-site observational study Methods Patients were included if they had undergone an elective procedure without sedation or anaesthesia in a cardiac catheterisation laboratory. Ambient room temperature was maintained between 18°C and 20°C. Passive warming with heated cotton blankets was applied. Nurses measured body temperature and thermal comfort before and after 342 procedures. Results Mean change in temperature was -0.08°C (Standard deviation 0.43). The reduction in temperature was more than 1°C after 11 procedures (3.2%). One patient whose temperature had reduced more than 1°C after their procedure reported thermal discomfort. A total of 12 patients were observed to be shivering post-procedure (3.6%). No demographic or clinical characteristics were associated with reduction in temperature of more than 1°C from pre to post-procedure. Conclusions Significant reduction in body temperature was rare in our sample of non-anaesthetised patients. Relevance to clinical practice Similar results would likely be found in other procedural contexts during procedures conducted in settings with comparable room temperatures where passive warming can also be applied with limited skin exposure.

Embedded PZT wafer sensors for structural health monitoring

Recent advances in structural integrity evaluation have led to the development. of PZT wafer sensors (PWAS) which can be embedded or surface mounted for both acoustic emission (AE) and ultrasonic (UT) modes, which forms an integrated approach for Structural Health Monitoring (SHM) of aerospace structures. For the fabrication of PWAS wafers, soft PZT formulation (SP-5H Grade containing dopants like BA, SM, CA, ZN, Y and HF) were used. The piezoelectric charge constant (d(33)) was measured by a d(33) meter. As a first step towards the final objective of developing Health monitoring methods with embedded PWAS, experiments were conducted on aluminum and composite plates of finite dimensions using PWAS sensors. The AE source was simulated by breaking 0.5mm pencil lead on the surface of a thin plate. Experiments were also conducted with surface mounted PZT films and conventional AE sensors in order to establish the sensitivity of PWAS. A comparison of results of theoretical and experimental work shows good agreement.