Incremental PCA algorithm for fringe pattern demodulation.

This work proposes a new algorithm for demodulating fringe patterns using principal component analysis (PCA). The algorithm is based on the incremental implantation of the singular value decomposition (SVD) technique for computing the principal values associated with a set of fringe patterns. Instead of processing an entire set of interferograms, the proposed algorithm proceeds in an incremental way, processing sequentially one (as minimum) interferogram at a given time. The advantages of this procedure are twofold. Firstly, it is not necessary to store the whole set of images in memory, and, secondly, by computing a phase quality parameter, it is possible to determine the minimum number of images necessary to accurately demodulate a given set of interferograms. The proposed algorithm has been tested for synthetic and experimental interferograms showing a good performance.

Robust fitting of Zernike polynomials to noisy point clouds defined over connected domains of arbitrary shape.

A new method for fitting a series of Zernike polynomials to point clouds defined over connected domains of arbitrary shape defined within the unit circle is presented in this work. The method is based on the application of machine learning fitting techniques by constructing an extended training set in order to ensure the smooth variation of local curvature over the whole domain. Therefore this technique is best suited for fitting points corresponding to ophthalmic lenses surfaces, particularly progressive power ones, in non-regular domains. We have tested our method by fitting numerical and real surfaces reaching an accuracy of 1 micron in elevation and 0.1 D in local curvature in agreement with the customary tolerances in the ophthalmic manufacturing industry.

Application of principal component analysis in phase-shifting photoelasticity.

Principal component analysis phase shifting (PCA) is a useful tool for fringe pattern demodulation in phase shifting interferometry. The PCA has no restrictions on background intensity or fringe modulation, and it is a self-calibrating phase sampling algorithm (PSA). Moreover, the technique is well suited for analyzing arbitrary sets of phase-shifted interferograms due to its low computational cost. In this work, we have adapted the standard phase shifting algorithm based on the PCA to the particular case of photoelastic fringe patterns. Compared with conventional PSAs used in photoelasticity, the PCA method does not need calibrated phase steps and, given that it can deal with an arbitrary number of images, it presents good noise rejection properties, even for complicated cases such as low order isochromatic photoelastic patterns.

Detection of hepatitis C virus (HCV) core-specific antibody suggests occult HCV infection among blood donors.

BACKGROUND: Blood transfusion safety is based on reliable donor screening for transmissible infections such as the hepatitis C virus (HCV) infection. STUDY DESIGN AND METHODS: A novel HCV core-specific antibody was assayed on random single donations from 2007 first-time blood donors who tested negative for anti-HCV and HCV RNA on routine screening. Sample collection broke the code between donations and donors for ethical reasons. RESULTS: Forty-two donations (2.1%) displayed reactivity in the novel test. The specificity of the reactivity was evaluated by a peptide inhibition assay, and testing against additional nonoverlapping HCV core peptide epitopes and other HCV antigens was performed on these samples. Six donations (14.3%; 0.30% from the total) were considered to contain anti-HCV after such supplemental testing. HCV RNA detection was also performed in peripheral blood mononuclear cells (PBMNCs) and serum or plasma samples from reactive donors after virus concentration by ultracentrifugation. HCV RNA tested negative in all PBMNCs samples, and a very low amount of viral genome was detected in serum or plasma concentrates from three anti-HCV core-reactive donors (7.1%) but not among concentrates from 100 randomly selected nonreactive donors. Sequencing of these polymerase chain reaction products revealed differences between the isolates that excluded partially sample contamination from a common source. CONCLUSION: These findings argue in favor of an ongoing occult HCV infection among these blood donors and account for some rather low, but perhaps not negligible, infection risk for such donations. Future studies involving larger samples of donations from traceable donors would enlighten the significance of these findings for the viral safety of the blood supply.

Impact of isolated hepatitis C virus (HCV) core-specific antibody detection and viral RNA amplification among HCV-seronegative dialysis patients at risk for infection.

Amplification of hepatitis C virus (HCV) RNA from blood detected occult HCV infections in 30.9% of 210 HCV-seronegative dialysis patients with abnormal liver enzyme levels that had evaded standard HCV testing practices. Isolated HCV core-specific antibody detection identified three additional anti-HCV screening-negative patients lacking HCV RNA amplification in blood who were considered potentially infectious. Together, these findings may affect management of the dialysis setting.

High prevalence of occult hepatitis C virus infection in patients with primary and secondary glomerular nephropathies.

The association of hepatitis C virus (HCV) infection and glomerulonephritis is well known. However, the relationship between immune-mediated glomerulonephritis and occult HCV, characterized by the presence of HCV-RNA in liver or in peripheral blood mononuclear cells in the absence of serological markers, is unknown. We tested this in 113 anti-HCV-negative patients; 87 with immune-mediated glomerulonephritis and 26 controls with hereditary glomerular nephropathies. All patients were serum HCV-RNA negative by conventional real-time PCR. Significantly, occult HCV-RNA (detectable viral RNA in peripheral blood mononuclear cells or in serum after ultracentrifugation) was found in 34 of 87 patients with immune-mediated glomerulonephritis versus 1 of 26 control patients. The serum creatinine levels were significantly higher in patients with immune-mediated glomerulonephritis with than in those without occult HCV (1.5 versus 1.1 mg/dl, respectively). A multivariate analysis adjusted for gender showed a significantly increased risk of occult HCV in patients with immune-mediated glomerulonephritis versus the controls (odds ratio of 13.29). Progression to end-stage renal disease tended to be faster in patients with immune-mediated glomerulonephritis and occult HCV than in the negative cases. Thus, occult HCV is strongly associated with immune-mediated glomerulonephritis and may have a role in the progression of the disease.

Fast two-dimensional simultaneous phase unwrapping and low-pass filtering.

Here, we present a fast algorithm for two-dimensional (2D) phase unwrapping which behaves as a recursive linear filter. This linear behavior allows us to easily find its frequency response and stability conditions. Previously, we published a robust to noise recursive 2D phase unwrapping system with smoothing capabilities. But our previous approach was rather heuristic in the sense that not general 2D theory was given. Here an improved and better understood version of our previous 2D recursive phase unwrapper is presented. In addition, a full characterization of it is shown in terms of its frequency response and stability. The objective here is to extend our previous unwrapping algorithm and give a more solid theoretical foundation to it.

Long-term virological follow up of patients with occult hepatitis C virus infection.

BACKGROUND: Patients with occult hepatitis C virus (HCV) infection (HCV-RNA in liver without detectable anti-HCV and serum HCV-RNA) may have viral RNA in peripheral blood mononuclear cells (PBMCs) and in serum after ultracentrifugation, and may present HCV-specific T-cell responses, but it is unknown whether these markers persist to be detectable over time. AIM: To perform a prospective virological long-term follow up of patients with occult HCV. METHODS: Viral markers were tested every 3-4 months during 55.7 ± 20.3 months in 37 patients with occult HCV who were under ursodeoxycholic acid treatment. RESULTS: Viral RNA was detectable in PBMCs of 31 patients during the follow up. In 23 of them, viral RNA in PBMCs was detected intermittently and in the other eight patients HCV-RNA was positive in a single sample. After ultracentrifugation, serum HCV-RNA was detected in 33 patients, being the viraemia intermittently detectable in 28, whereas in the remaining five patients, serum HCV-RNA was positive only once. Only one patient tested always HCV-RNA negative in PBMCs and in ultracentrifuged serum during follow up. Specific Core, NS3, and/or NS4 T-cell responses were found in 31 of the patients. The patient who was always HCV-RNA negative in PBMCs and in ultracentrifuged serum had specific HCV-T-cell responses. CONCLUSIONS: Occult HCV infection persists over time with fluctuating viraemia levels that induce and maintain specific T-cell responses against viral proteins.

Noise robust linear dynamic system for phase unwrapping and smoothing.

Phase unwrapping techniques remove the modulus ambiguities of wrapped phase maps. The present work shows a first-order feedback system for phase unwrapping and smoothing. This system is a fast sequential unwrapping system which also allows filtering some noise because in deed it is an Infinite Impulse Response (IIR) low-pass filter. In other words, our system is capable of low-pass filtering the wrapped phase as the unwrapping process proceeds. We demonstrate the temporal stability of this unwrapping feedback system, as well as its low-pass filtering capabilities. Our system even outperforms the most common and used unwrapping methods that we tested, such as the Flynn's method, the Goldstain's method, and the Ghiglia least-squares method (weighted or unweighted). The comparisons with these methods shows that our system filters-out some noise while preserving the dynamic range of the phase-data. Its application areas may cover: optical metrology, synthetic aperture radar systems, magnetic resonance, and those imaging systems where information is obtained as a demodulated wrapped phase map.

Deflectometric method for the measurement of user power for ophthalmic lenses.

This paper presents a deflectometric technique to measure the power of an ophthalmic lens as perceived by the user. It is based on a calibrated camera acting as a pinhole in order to measure ray deflection along the same path as the visual axis when the lens is held in front of the eye. We have analyzed numerically the accuracy of our technique, and it has been compared experimentally with a commercial "lens mapper" and with the real user power calculated from the measured topography of the lens surfaces to state the reliability and accuracy of the presented technique.

A self-tuning phase-shifting algorithm for interferometry.

In Phase Stepping Interferometry (PSI) an interferogram sequence having a known, and constant phase shift between the interferograms is required. Here we take the case where this constant phase shift is unknown and the only assumption is that the interferograms do have a temporal carrier. To recover the modulating phase from the interferograms, we propose a self-tuning phase-shifting algorithm. Our algorithm estimates the temporal frequency first, and then this knowledge is used to estimate the interesting modulating phase. There are several well known iterative schemes published before, but our approach has the unique advantage of being very fast. Our new temporal carrier, and phase estimator is capable of obtaining a very good approximation of their temporal carrier in a single iteration. Numerical experiments are given to show the performance of this simple yet powerful self-tuning phase shifting algorithm.

Identification of serologically silent occult hepatitis C virus infection by detecting immunoglobulin G antibody to a dominant HCV core peptide epitope.

BACKGROUND/AIMS: Occult HCV infection has been described among anti-HCV-HCV RNA-negative individuals with abnormal transaminase values in whom HCV RNA is detected in liver. METHODS: IgG antibody to an HCVcore-derived peptide (anti-HCVcore) was investigated in 145 patients with serologically silent occult HCV infection. RESULTS: At the time of the diagnostic biopsy 45/145 (31%) occult HCV-infected patients tested IgG anti-HCVcore-positive but none of the 140 patients with HCV-unrelated liver disease (P<0.001). Among 23 IgG anti-HCVcore-positive patients at baseline, 22 remained antibody-reactive (one became antibody-negative). Similarly, 17/31 baseline anti-HCVcore-negative patients remained non-reactive whereas 14 seroconverted to IgG anti-HCVcore (although transiently in 10 patients). Thus, a total of 59/145 (40.7%) patients with occult HCV infection showed IgG anti-HCVcore reactivity at any time point analyzed, including 14 initially non-reactive patients. By supplemental immunoblot assay 16 sera reacted weakly with an HCVcore-peptide band (indeterminate result) of which 10 (62.5%) reacted in the IgG anti-HCVcore assay. Occult HCV-infected patients who tested anti-HCVcore-positive showed more frequently signs of necro-inflammation (P=0.035) and greater percentages of HCV RNA-positive hepatocytes (P=0.004) compared with those anti-HCVcore-negative. CONCLUSIONS: This work documents that IgG anti-HCVcore testing identifies occult HCV infection among seronegative, non-viremic patients using screening tests and may be useful in tracking anti-HCV-negative infections.

Serum immunoglobulin G antibodies to the GOR autoepitope are present in patients with occult hepatitis C virus (HCV) infection despite lack of HCV-specific antibodies.

Antibody responses to the GOR autoepitope are frequently detected among anti-hepatitis C virus (anti-HCV)-positive patients with chronic hepatitis. Sera from 110 anti-HCV-negative patients with occult HCV infection, as diagnosed by detection of HCV RNA in hepatic tissue, were investigated for GOR antibody reactivity. A positive test for anti-GOR immunoglobulin G (IgG) was found for 22 (20%) of them. The frequency and titers of anti-GOR IgG were significantly lower than those in chronic hepatitis C patients (70/110, 63.6%; P < 0.001). Anti-GOR IgG was not detected in any of the 120 patients with HCV-unrelated liver disease. The anti-GOR IgG assay showed specificity and sensitivity values of 100% and 20%, respectively, among the sera from patients with occult HCV infection; the positive and negative predictive values were 100% and 44.3%, respectively. None of the clinical, laboratory, or histological characteristics of the patients with occult HCV infection were different according to GOR antibody status, except that the percentage of HCV RNA-positive hepatocytes was significantly greater (P = 0.042) in patients with occult HCV infection who tested positive for anti-GOR IgG. In conclusion, serum anti-GOR IgG is present in patients with occult HCV infection, despite a lack of detectable HCV-specific antibodies as determined by commercial tests. Testing for anti-GOR IgG in patients in whom HCV RNA is not detected in their sera may help with the identification of a subset of patients with occult HCV infection without the need to perform a liver biopsy.

Cellular immune responses associated with occult hepatitis C virus infection of the liver.

Occult hepatitis C virus (HCV) infection is a type of recently identified chronic infection that is evidenced only by detection of HCV RNA in liver; patients consistently test negative for antibodies to HCV and HCV RNA in serum. Using ex vivo and in vitro measures of T-cell responses, we have identified functional virus-specific memory CD4(+) and CD8(+) T cells in the peripheral blood of patients with occult HCV infection. The features of the virus-specific T cells were consistent with immune surveillance functions, supporting previous exposure to HCV. In addition, the magnitudes of CD4(+) and CD8(+) T-cell responses were in parallel and correlated inversely with the extent of liver HCV infection. The detection of HCV-specific T cells in individuals in whom HCV RNA can persist in the liver despite the absence of viremia and antibodies indicates that HCV replication is prolonged in the face of virus-specific CD4(+) and CD8(+) T-cell responses. These findings demonstrate that HCV-specific cellular immune responses are markers not only of previous exposure to and recovery from HCV but also of ongoing occult HCV infection.

Combined hepatitis C virus (HCV) antigen-antibody detection assay does not improve diagnosis for seronegative individuals with occult HCV infection.

A combined hepatitis C virus (HCV) antigen-antibody assay was evaluated for 115 seronegative individuals with occult HCV infection. The assay was reactive in one patient and negative to weakly reactive in three others (all four gave indeterminate results by supplemental assay) but failed to detect HCV in the remaining patients. Despite increased sensitivity the combined assay does not improve serodiagnosis of occult HCV infection.

Virus-specific T-cell responses associated with hepatitis C virus (HCV) persistence in the liver after apparent recovery from HCV infection.

Hepatitis C virus (HCV) RNA persistence in the liver has been described even after apparent resolution of HCV infection. Because T-cell reactivity plays a role in recovery from HCV infection, virus-specific T-cell responses were investigated in apparently recovered individuals in whom hepatic HCV RNA persistence was documented: 15 sustained virological responders to interferon (IFN)-treatment and 9 asymptomatic aviremic anti-HCV carriers. HCV-specific CD4(+) T-cell proliferative responses were detected significantly more often in apparently recovered individuals (sustained virological responders: 60%; asymptomatic anti-HCV carriers: 66%) compared with 50 chronic hepatitis C patients (28%; P < 0.05). However, T-cell frequencies and numbers tended to decline over time and the number of HCV proteins targeted by CD4(+) T-cell proliferative responses was limited. Interestingly, liver viral load correlated inversely with virus-specific immune responses. Thus, CD4(+) T-cell responders showed significantly lower hepatic HCV RNA levels (P < 0.05). HCV-specific IFN-gamma-secreting CD4(+) T-cells were not detected in all the apparently recovered patients although they were found significantly more often compared with chronic hepatitis C patients (P < 0.05). Also, HCV NS3-specific CD8(+) T-cells were detected in 11 HLA-A2-positive apparently recovered individuals (8 sustained virological responders and 3 asymptomatic anti-HCV carriers); T-cell frequencies tended to be greater in those patients who had lower hepatic viral levels. In conclusion, HCV-specific T-cells are detectable in apparently recovered individuals in whom HCV RNA can persist in the liver indicating that HCV replication may be prolonged in the face of an insufficient or inadequate virus-specific CD4(+) and CD8(+) T-cell response.

Evidence for a relation between the viral load and genotype and hepatitis C virus-specific T cell responses.

BACKGROUND/AIMS: The reason why patients with hepatitis C virus (HCV) genotype non-1 infection respond better to antiviral therapy than patients with genotype 1 infection is not known. The aim of this study is to explore the relation between the viral genotype, viral load, and the endogenous T cell response. METHODS: The viral genotype, the viral load, and the endogenous proliferative T cell response to the non-structural 3 protein (NS3) was analysed using serum and peripheral blood mononuclear cells from 103 patients with chronic HCV infection. RESULTS: Among 71 nontreated patients a T cell response was more common among those infected by genotype 3, as compared to those infected with genotype 1 (P<0.05). Among 32 patients undergoing antiviral therapy, presence of a T cell response was more common in genotype non-1 infected patients than in those infected by genotype 1 (P<0.01). Presence of a T cell response was related to a more rapid viral clearance (P<0,05), a negative HCV RNA test at week 12 (P<0.05), and a shorter viral half-life (P<0.05). CONCLUSIONS: The presence of an NS3-specific T cell response is related to the viral genotype and to a more rapid clearance of HCV RNA during antiviral therapy.

Virus-specific effector CD4+ T-cell responses in hemodialysis patients with hepatitis C virus infection.

Patients with chronic renal failure undergoing hemodialysis who are infected with hepatitis C virus (HCV) may test consistently anti-HCV negative. Because CD4(+) T-cells provide help for antibody production virus-specific effector CD4(+) T-cell responses were investigated in relation to anti-HCV positivity in 15 hemodialysis patients grouped according to HCV antibody and viremia. CD4(+) T-cell reactivity was studied in peripheral blood mononuclear cells by standard lymphocyte proliferation assay and phenotypic/functional characterization (cell-surface staining/cytokine secretion) by flow cytometry. HCV-specific CD4(+) T-cell proliferation in viremic hemodialysis patients was weak or absent independently of their anti-HCV status. Virus-specific CD4(+) T-cells displayed a memory phenotype and showed low to undetectable capacity to secrete effector interferon (IFN)-gamma. Impaired activation-induced cytokine secretion appeared to be Th1 (IFN-gamma) but not Th2 (interleukin-4)-directed and was virus-specific as cytomegalovirus responses were preserved. The frequency ex vivo of CD3(+)CD4(+)IFN-gamma(+) T-cells was independent of the HCV antibody status and comparable between viremic (range: 0.08-1.54%) or non-viremic (0.11-3.2%) hemodialysis patients and healthy donors (0.13-1.10%; P = 0.58). The numbers of CD3(+)CD4(+)IFN-gamma(+) T-cells augmented slightly (P = 0.047) in HCV-infected hemodialysis patients but markedly in only one (greater than ninefold) after HCV stimulation. In conclusion, hemodialysis patients show limited HCV-specific effector CD4(+) Th1-cell responses which nonetheless seem unrelated to the anti-HCV status and are not more impaired due to the ongoing hemodialysis.

Features of the CD4+ T-cell response in liver and peripheral blood of hepatitis C virus-infected patients with persistently normal and abnormal alanine aminotransferase levels.

BACKGROUND/AIMS: The liver is the primary site of hepatitis C virus (HCV) replication; intrahepatic T-cell responses may influence liver disease severity. METHODS: HCV-specific CD4(+) T-cell reactivity was investigated ex vivo in paired liver tissue and peripheral blood from 42 chronic HCV patients. RESULTS: The frequencies with which HCV-specific HLA class-II-restricted CD4(+) T-cell proliferation were observed were 29% in liver and 36% in peripheral blood. Among responses, non-structural-3 protein (NS3)-specific T-cell proliferation was dominant but non-exclusive and did rarely occur concurrently in liver infiltrate and peripheral blood suggesting liver compartmentalization of a CD4(+) T-cells population. Compared with 24 patients with abnormal ALT levels, 18 HCV carriers with persistently normal ALT levels had similar serum and liver viral loads but showed: (i) a low-activity grade and stage chronic hepatitis (P<0.001); (ii) less intrahepatic CD4(+) T-lymphocytes (P<0.01); (iii) less frequent intrahepatic (17 vs. 33%) and peripheral (17 vs. 38%) NS3-specific CD4(+) T-cell proliferation; (iv) less often in vitro T-helper (Th)1 (interferon-gamma) cytokine production (2 vs. 18%; P<0.001). CONCLUSIONS: Our data show a low frequency of intrahepatic HCV-specific HLA class-II-restricted CD4(+) Th1 responses in patients with chronic HCV. However, these Th1 responses are detected more often in those patients with overt clinical and histological disease.