Association of CCR5Δ32 Deletion and Human Cytomegalovirus Infection With Colorectal Cancer in Tunisia

Background and objectives: Human cytomegalovirus (HCMV) and genetic polymorphisms of the chemokine receptor 5 have been suggested as factors associated with the progression of colorectal cancer (CRC). The aim of the study was to evaluate the associations of both CCR5Δ32 genetic deletion and/or HCMV virus infection with CRC in Tunisia. Materials and methods: The association between HCMV and CRC was validated by Nested PCR technology performed for HCMV and HCMV-specific serum IgG and IgM antibodies were investigated by enzyme-linked immunosorbent assay. Experiments were carried out on 40 tumor and 35 peri-tumor tissues, 100 blood from CRC patients and on 140 blood samples from healthy subjects and finaly serum samples of 80 patients with CRC and 100 healthy individuals. A conventional PCR has been optimized for the detection of CCR5Δ32 in100 CRC patients and 100 healthy subjects. Results: Our results show that HCMV is significantly active in 93% of patients compared to 60% in controls (p < 0.0001, OR = 8.85, 95% CI: 3.82 -20.50). Compared to the healthy controls, the titers of IgG and IgM antiCMV antibodies in CRC patients were significantly higher than in healthy subjects (p value < 0,0001 for IgG and IgM). Statistical analysis revealed a lack of association between CCR5Δ32 mutation and colorectal cancer (p = 0.788, OR = 1.265, 95% CI: 0.228-7.011). Conclusion: our data confirmed that the HCMV infection was related to the development of CRC and that CRC cells may be infected more favorably by HCMV. Given the importance of the CCR5 in inflammation and therefore CRC progression, further studies still needed to evaluate CCR5 role as a potential candidate gene for CRC susceptibility under other polymorphisms.

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Clinical Significance of Annexin A2 Expression in Breast Cancer Patients

Cancers ◽

10.3390/cancers13010002 ◽

2020 ◽

Vol 13 (1) ◽

pp. 2

Author(s):

Lee D. Gibbs ◽

Kelsey Mansheim ◽

Sayantan Maji ◽

Rajesh Nandy ◽

Cheryl M. Lewis ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Patients ◽

Clinical Significance ◽

Cell Lines ◽

Breast Cancer Subtypes ◽

Enzyme Linked Immunosorbent Assay ◽

Breast Cancer Patients ◽

Serum Samples ◽

Cancer Subtypes ◽

Tumor Tissues

Increasing evidence suggests that AnxA2 contributes to invasion and metastasis of breast cancer. However, the clinical significance of AnxA2 expression in breast cancer has not been reported. The expression of AnxA2 in cell lines, tumor tissues, and serum samples of breast cancer patients were analyzed by immunoblotting, immunohistochemistry, and enzyme-linked immunosorbent assay, respectively. We found that AnxA2 was significantly upregulated in tumor tissues and serum samples of breast cancer patients compared with normal controls. The high expression of serum AnxA2 was significantly associated with tumor grades and poor survival of the breast cancer patients. Based on molecular subtypes, AnxA2 expression was significantly elevated in tumor tissues and serum samples of triple-negative breast cancer (TNBC) patients compared with other breast cancer subtypes. Our analyses on breast cancer cell lines demonstrated that secretion of AnxA2 is associated with its tyrosine 23 (Tyr23) phosphorylation in cells. The expression of non-phosphomimetic mutant of AnxA2 in HCC1395 cells inhibits its secretion from cells compared to wild-type AnxA2, which further suggest that Tyr23 phosphorylation is a critical step for AnxA2 secretion from TNBC cells. Our analysis of AnxA2 phosphorylation in clinical samples further confirmed that the phosphorylation of AnxA2 at Tyr23 was high in tumor tissues of TNBC patients compared to matched adjacent non-tumorigenic breast tissues. Furthermore, we observed that the diagnostic value of serum AnxA2 was significantly high in TNBC compared with other breast cancer subtypes. These findings suggest that serum AnxA2 concentration could be a potential diagnostic biomarker for TNBC patients.

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Correlation between IL-10 as Cancer Biomarker and Demographic Characteristics of Colorectal Cancer Patients

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i26b31477 ◽

2021 ◽

pp. 1-10

Author(s):

Rahin Sh Hamad ◽

Bushra H. Shnawa ◽

Shereen J. Al-Ali

Keyword(s):

Colorectal Cancer ◽

Pearson Correlation ◽

Serum Levels ◽

Interleukin 10 ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

High Morbidity ◽

Serum Samples ◽

Cancer Pathogenesis ◽

Colorectal Cancer Patients

Colorectal cancer (CRC) is classified as one of the most prevalent cancer types worldwide, with high morbidity and mortality rates. Patients of CRC have been shown to express a detectable cytokine in serum which contributes to cancer pathogenesis. Therefore, the serum interleukin 10 (IL-10) level in CRC patients was investigated in this study. Patients' medical records with CRC admitted to the Rizgary and Nanakali hospitals, Erbil, Iraq was analyzed as the study group compared to the healthy volunteers' control group. Seventy-one serum samples were collected, thirty-one from diagnosed CRC patients and forty from healthy controls. The concentrations of IL-10 in the sera were assessed by enzyme-linked immunosorbent assay (ELISA). The present finding showed that IL-10 Was significantly elevated in CRC patients' sera compared to the control group, suggesting confirmation of its usefulness for detecting CRC patients' prognosis. A non-significant Pearson correlation was detected between IL-10 serum levels and the CRC group's age, gender, and body mass index. Herein is the first study on the evaluation of IL-10 levels in CRC patients in Kurdistan, Iraq.

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Different expression pattern of human cytomegalovirus-encoded microRNAs in circulation from virus latency to reactivation

Journal of Translational Medicine ◽

10.1186/s12967-020-02653-w ◽

2020 ◽

Vol 18 (1) ◽

Author(s):

Wanqing Zhou ◽

Cheng Wang ◽

Meng Ding ◽

Yuying Bian ◽

Yujie Zhong ◽

...

Keyword(s):

Human Cytomegalovirus ◽

Characteristic Curve ◽

Antiviral Treatment ◽

Serum Levels ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Prediction Ability ◽

Stem Loop ◽

Virus Latency ◽

Polymerase Chain

Abstract Background Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness. Methods Serum samples from 59 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sample. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease prediction ability of differential expression HCMV-miRNAs during the antiviral treatment. Results The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miR-UL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P < 0.01). Receiver operating characteristic curve (ROC) analysis were performed on the selected HCMV-miRNAs in all of the patients and controls that enrolled in this study, and which ranged from 0.72 to 0.80 in the autoimmune patients. In addition, hcmv-miR-US25-1-3p levels were significantly correlated with HCMV DNA load (r = 0.349, P = 0.007), and were obviously higher in the reactivation set than the latency set in the autoimmune patients, which could be a predictor for the monitoring of the antiviral treatment. Conclusions HCMV miRNAs profile showed markedly shift-switch from latency to reactivation in circulation from HCMV infected patients and hcmv-miR-US25-1-3p may be served as a predictor for the switch upon reactivation from latency in patients suffered with autoimmune diseases.

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Evaluation of serum CXCL5 as a serum marker for prognosis of colorectal cancer patients.

Journal of Clinical Oncology ◽

10.1200/jco.2011.29.4_suppl.442 ◽

2011 ◽

Vol 29 (4_suppl) ◽

pp. 442-442 ◽

Cited By ~ 1

Author(s):

M. Kawamura ◽

Y. Toiyama ◽

K. Tanaka ◽

H. Yasuda ◽

H. Fujikawa ◽

...

Keyword(s):

Colorectal Cancer ◽

Overall Survival ◽

Liver Metastasis ◽

Cancer Patients ◽

Normal Serum ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Serum Samples ◽

Preoperative Serum ◽

Colorectal Cancer Patients

442 Background: CXCL5 is known as CXC chemokine which promotes angiogenesis related to cancer. However, the function of serum level of CXCL5 (sCXCL5) has not been fully studied in colorectal cancer. The purpose of this study was to evaluate the relationships between preoperative sCXCL5 and clinicopathological features and prognosis in colorectal cancer. Methods: This was a single-institution, retrospective study. Preoperative serum samples of 250 colorectal cancer patients (between 1998 and 2007, median age: 65.3 years, male 159/female 91) were available for the study, and 33 normal serum was examined and used as a control. sCXCL5 level was assayed using a commercially available enzyme-linked immunosorbent assay kit, and analyzed statistically. Results: Mean level of sCXCL5 was significantly higher in colorectal cancer patients than in control group (p=0.013). Patients with liver metastases had significantly higher sCXCL5 level than those without metastases (p=0.0086), and in logistic analysis, sCXCL5 was an independent marker for predicting liver metastasis (p=0.040). Overall survival of patients with elevated sCXCL5 level was significantly worse than those with lower sCXCL5 (p=0.0006). Conclusions: Preoperative sCXCL5 level was increased in colorectal cancer patients compared to in healthy volunteer and elevated sCXCL5 was correlated with liver metastasis and poor prognosis for overall survival in colorectal cancer patients. Elevated sCXCL5 has been proposed as a useful predictive marker for liver metastasis and overall survival in colorectal cancer. No significant financial relationships to disclose.

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Molecular Fine-Specificity Analysis of Antibody Responses to Human Cytomegalovirus and Design of Novel Synthetic-Peptide-Based Serodiagnostic Assays

Journal of Clinical Microbiology ◽

10.1128/jcm.37.1.179-188.1999 ◽

1999 ◽

Vol 37 (1) ◽

pp. 179-188 ◽

Cited By ~ 30

Author(s):

Astrid E. Greijer ◽

Jos M. G. van de Crommert ◽

Servi J. C. Stevens ◽

Jaap M. Middeldorp

Keyword(s):

Human Cytomegalovirus ◽

Immunoglobulin M ◽

Antibody Responses ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Peptide Epitopes ◽

Fine Specificity ◽

Marker Proteins ◽

Highly Sensitive ◽

Virion Antigen

To identify single immunodominant marker proteins which can replace complex virion antigen in serodiagnostic assays, we investigated in detail the molecular fine specificity of antibody responses in different individuals with latent or active human cytomegalovirus (HCMV) infection. An overview of the HCMV proteins recognized by human antibodies was obtained by immunoblotting. For selected immunodominant proteins the epitope fine specificity of the antibody response was determined by a peptide-scanning enzyme-linked immunosorbent assay (ELISA). Epitope clusters were synthesized as combination peptides and were used for further serologic analysis of immunoglobulin M (IgM) and IgG reactivities with panels of sera from different groups of patients in comparison to those with cytomegalovirus (CMV) virion antigen. Several serum samples had significantly higher reactivities with peptides than with the CMV virion antigen. However, individual serum samples occasionally recognized diverse peptide epitopes, stressing the importance of using combinations of peptides in serologic assays. From these studies we were able to define a specific combination of peptides derived from pp52 (UL44) and pp150 (UL32) for the specific and highly sensitive early detection of HCMV IgM, whereas a combination of peptides from pp150 (UL32), gB (UL55), and pp28 (UL99) was selected to give optimal and specific reactivity with HCMV IgG. On the basis of the results obtained with these peptide combinations, new, highly specific serodiagnostic assays were constructed. These assays had sensitivities of 98.9 and 96.4% for IgG and IgM, respectively, in comparison with the results obtained with the “gold standard,” the virion antigen-based ELISA. From the results of this study we conclude that specific combinations of highly defined synthetic peptides can replace complex HCMV virion extracts used in current serodiagnostics and may add to further standardization of HCMV serology.

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Detection of Immunoglobulin G Antibodies to Neospora caninum in Humans: High Seropositivity Rates in Patients Who Are Infected by Human Immunodeficiency Virus or Have Neurological Disorders

Clinical and Vaccine Immunology ◽

10.1128/cvi.13.1.84-89.2006 ◽

2006 ◽

Vol 13 (1) ◽

pp. 84-89 ◽

Cited By ~ 59

Author(s):

Janaína Lobato ◽

Deise A. O. Silva ◽

Tiago W. P. Mineo ◽

Jodi D. H. F. Amaral ◽

Gesmar R. Silva Segundo ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Immunoglobulin G ◽

Neurological Disorders ◽

Healthy Subjects ◽

Neospora Caninum ◽

Fluorescent Antibody ◽

Antibody Test ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Immunodeficiency Virus

ABSTRACT Considering that little is known about the epidemiology of Neospora caninum infection in humans, particularly in populations with high Toxoplasma gondii infection rates, the present study aimed to investigate the presence of antibodies to N. caninum in T. gondii-seropositive and -seronegative individuals. A total of 256 serum samples divided into four groups (61 samples from human immunodeficiency virus [HIV]-positive patients, 50 samples from patients with neurological disorders, 91 samples from newborns, and 54 samples from healthy subjects) were assessed for N. caninum and T. gondii serologies by indirect fluorescent-antibody test, enzyme-linked immunosorbent assay, and immunoblotting (IB). Immunoglobulin G antibodies to N. caninum were predominantly detected in HIV-infected patients (38%) and patients with neurological disorders (18%), while newborns and healthy subjects showed lower seropositivity rates (5% and 6%, respectively). Seropositivity to N. caninum was significantly associated with seropositivity to T. gondii in both HIV-infected patients and patients with neurological disorders. Seroreactivity to N. caninum was confirmed by IB, with positive sera predominantly recognizing the 29-kDa antigen of N. caninum. The results of this study indicate the presence of N. caninum infection or exposure in humans, particularly in HIV-infected patients or patients with neurological disorders, who could have opportunistic and concurrent infections with T. gondii. These findings may bring a new concern for the unstable clinical health of HIV-infected patients and the actual role of N. caninum infection in immunocompromised patients.

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Serum Expression of β-Catenin Is a Potential Detection Marker in Patients with Colorectal Cancer

Disease Markers ◽

10.1155/2019/5070524 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Shue Li ◽

Mao Huang ◽

Qiao Liu ◽

Ding Wang ◽

Rui Wu ◽

...

Keyword(s):

Colorectal Cancer ◽

Roc Curve ◽

Disease Process ◽

Disease Diagnosis ◽

Colorectal Disease ◽

Clinicopathological Parameters ◽

Enzyme Linked Immunosorbent Assay ◽

Potential Candidate ◽

Diagnostic Efficiency ◽

Significant Difference

Object. To investigate the correlation between the level of serum β-catenin and the disease progression of colorectal polyp (CRP) and colorectal cancer (CRC) and find its potential diagnostic value. Methods. A total of 327 clinical serum samples and their electronic medical records were collected. Detecting by enzyme-linked immunosorbent assay (ELISA), the correlations of serum β-catenin with tumor marker carcinoembryonic antigen (CEA) and CRC clinicopathological parameters and the receiver operating characteristic (ROC) curve were analyzed. Results. Serum β-catenin levels in the CRP and CRC patients were significantly higher than those in the healthy control (HC) group (P<0.05 and P<0.001). Compared with CRP, serum β-catenin level in CRC was also increased (P<0.05). However, there was no significant difference in gender, age, location, tumor size, Dukes staging, or metastasis (P>0.05) between serum β-catenin and clinical parameters of CRC. There was no correlation between serum β-catenin levels and CEA in CRC patients (P=0.14). ROC curve analysis showed that serum β-catenin possessed the maximum diagnostic efficiency in CRP (AUC=0.73, P<0.05) with 86.41% sensitivity and 51.56% specificity. β-Catenin combined with CEA had the highest diagnostic efficiency (AUC=0.88, P<0.05) with 81.88% sensitivity and 73.44% specificity. With CRC patients from CRP patients, ROC analysis of the combining detection (AUC=0.70, P<0.05) had the 70% sensitivity and 84.5% specificity. Conclusion. The serum β-catenin levels are gradually increased in CRP and CRC, while there is no correlation between its levels and CRC disease process. Single serum β-catenin or combined CEA would be one of the potential candidate biomarkers for colorectal disease diagnosis.

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Endosialin (TEM1) as a Diagnostic, Progression, and Prognostic Serum Marker for Patients With Colorectal Cancer—A Preliminary Study

Cancer Control ◽

10.1177/1073274820903351 ◽

2020 ◽

Vol 27 (1) ◽

pp. 107327482090335

Author(s):

Łukasz Pietrzyk ◽

Paulina Wdowiak

Keyword(s):

Colorectal Cancer ◽

Serum Concentration ◽

Early Stage ◽

Serum Marker ◽

Carbohydrate Antigen ◽

Stage I ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Individuals ◽

Serum Samples ◽

The Mean

Colorectal cancer (CRC) is one of the most common cancers worldwide usually diagnosed in the advanced stage. In this study, the serum concentration of tumor endothelial marker 1 (TEM1) was measured and correlated with clinicopathological features to evaluate whether TEM1 might serve as a biomarker for early CRC diagnosis, progression, and prognosis. The concentration of TEM1 was measured in the serum samples of 45 patients with CRC and 35 healthy individuals using enzyme-linked immunosorbent assay test. The mean serum concentration of TEM1 was significantly higher in the patients with CRC compared to the healthy individuals (1.31 ± 0.16 vs 0.92 ± 0.90 ng/mL; P < .001). The mean concentration of TEM1 significantly increased in the patients having CRC with early stage (stage I + II) compared to noncancer control individuals (stage I + II vs control 1.21 ± 0.13 ng/mL: 0.92 ± 0.90 ng/mL; P < .001). The TEM1 concentration in blood serum also showed a significant association with the development of T stages ( P < .001), N stages ( P < .001), and M stages ( P = .006). The TEM1 sensitivity and specificity in CRC detection are higher than routinely used blood markers (carcinoembryonic antigen [CEA] and carbohydrate antigen [Ca 19-9]). Patients with high TEM1 concentration (≥1.055 ng/mL) had a worse overall survival rate compared to the patients having CRC with low TEM1 concentration (<1.055 ng/mL). In conclusion, TEM1 can act as a potential diagnostic, progression, and prognostic serum biomarker for patients with CRC; TEM1 might be a good supplement for commonly used markers CEA and Ca 19-9.

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Use of the 27-Kilodalton Recombinant Protein fromParacoccidioides brasiliensis in Serodiagnosis of Paracoccidioidomycosis

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.6.826-830.1998 ◽

1998 ◽

Vol 5 (6) ◽

pp. 826-830 ◽

Cited By ~ 28

Author(s):

B. L. Ortiz ◽

S. Díez ◽

M. E. Urán ◽

J. M. Rivas ◽

M. Romero ◽

...

Keyword(s):

Recombinant Protein ◽

Healthy Subjects ◽

Paracoccidioides Brasiliensis ◽

Cell System ◽

Etiologic Agent ◽

Immunological Methods ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

High Coverage ◽

Cross Reactions

ABSTRACT Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in Latin America; it is usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods. Although the latter are effective, two circumstances, cross-reactions with other mycotic agents and antigen preparation that is marked by extreme variability among lots, hinder proper standardization of the procedures. To circumvent this lack of reproducibility, molecular biology tools were used to produce a recombinant 27-kDa-molecular-mass antigen from this fungus; a sizable quantity of this antigen was obtained through fermentation of Escherichia coli DH5α, which is capable of expressing the fungal protein. The latter was purified by the Prep-Cell System (Bio-Rad); the recovery rate of the pure protein was approximately 6%. A battery of 160 human serum samples, consisting of 64 specimens taken at the time of diagnosis from patients with PCM representing the various clinical forms plus 15 serum specimens each from patients with histoplasmosis and aspergillosis, 10 each from patients with cryptococcosis and tuberculosis, 6 from patients with coccidioidomycosis, and 40 from healthy subjects, were all tested by an indirect enzyme-linked immunosorbent assay with the purified 27-kDa recombinant protein. The latter was used at a concentration of 1.0 μg/well; there were three serum dilutions (1:1,000, 1:2,000, and 1:4,000). The experiment was repeated at least twice. The average sensitivity for both experiments was 73.4%; in comparison with the healthy subjects, the specificity for PCM patients was 87.5% while for patients with other mycoses, it was 58.7%. Important cross-reactions with sera from patients with aspergillosis and histoplasmosis were detected. The positive predictive value of the test was 90.4%. These results indicate that it is possible to employ recombinant antigenic proteins for the immunologic diagnosis of PCM and, by so doing, achieve high coverage rates. Furthermore, antigen reproducibility can now be ensured, thus facilitating inter- and intralaboratory standardization.

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First direct assay for intact human proinsulin

Clinical Chemistry ◽

10.1093/clinchem/44.7.1514 ◽

1998 ◽

Vol 44 (7) ◽

pp. 1514-1519 ◽

Cited By ~ 21

Author(s):

Paule Houssa ◽

Bo Dinesen ◽

Michelle Deberg ◽

Bruce H Frank ◽

Chris Van Schravendijk ◽

...

Keyword(s):

Healthy Subjects ◽

Limit Of Detection ◽

Insulin Dependent Diabetes Mellitus ◽

Dependent Diabetes Mellitus ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

C Peptide ◽

Human Proinsulin ◽

The Mean ◽

A Chain

Abstract We describe a sensitive two-site sandwich enzyme-linked immunosorbent assay for the measurement of intact human proinsulin in 100 μL of serum or plasma. The assay is based on the use of two monoclonal antibodies specific for epitopes at the C-peptide/insulin A chain junction and at the insulin B chain/C-peptide junction, respectively. Cross-reactivities with insulin, C-peptide, and the four proinsulin conversion intermediates were negligible. The detection limit in buffer was 0.2 pmol/L (3 standard deviations from zero). The working range was 0.2–100 pmol/L. The mean intra- and interassay coefficients of variation were 2.4% and 8.9%, respectively. The mean recovery of added proinsulin was 103%. Dilution curves of 40 serum samples are parallel to the proinsulin calibration curve. Proinsulin concentrations in 20 fasting healthy subjects were all above the limit of detection: median (range), 2.7 pmol/L (1.1–6.9 pmol/L). Six fasting non-insulin-dependent diabetes mellitus and five insulinoma patients had proinsulin concentrations significantly higher than healthy subjects: median (range), 7.7 pmol/L (3.2–18 pmol/L) and 153 pmol/L (98–320 pmol/L), respectively.

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