Proteomic Analysis ofTrichinella spiralisMuscle Larval Excretory-Secretory Proteins Recognized by Early Infection Sera

Although the excretory-secretory (ES) proteins ofTrichinella spiralismuscle larvae are the most commonly used diagnostic antigens for trichinellosis, their main disadvantage is the false negative results during the early stage of infection and cross-reaction of their main components (43, 45, 49, and 53 kDa) with sera of patients with other helminthiasis. The aim of this study was to identify early specific diagnostic antigens inT. spiralisES proteins with 30–40 kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66 kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30–40 kDa were recognized and identified by MALDI-TOF/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. These proteins might be the early specific diagnostic antigens for trichinellosis.

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Identification of a host collagen inducing factor from the excretory secretory proteins ofTrichinella spiralisusing immunoscreening

10.1101/319483 ◽

2018 ◽

Author(s):

Mi Kyung Park ◽

Hae-Jin Kim ◽

Min Kyoung Cho ◽

Shin Ae Kang ◽

So Young Park ◽

...

Keyword(s):

Signaling Pathway ◽

Serine Proteases ◽

Collagen Synthesis ◽

Trichinella Spiralis ◽

Expression System ◽

Molecular Characteristics ◽

Secretory Proteins ◽

Collagen Gene ◽

Smad Signaling

AbstractBackgroundIn a previous study, we found thatTrichinella spiralisexcretory and secretory proteins (ES-P) most likely activate collagen synthesis via TGF-β/Smad signaling, and this event could influence collagen capsule formation.Methodology/Principal FindingsIn order to identify the specific collagen inducing factor, ES-P was fractionated by a Superdex 200 10/300 GL column. We obtained three large fractions, F1, F2, and F3, but only F3 had collagen gene inducing ability. After immunoscreening, 10 collagen inducing factor candidates were identified. Among them, TS 15-1 and TS 15-2 were identical to the putative trypsin ofT. spiralis. The deduced TS 15-1 (M.W. = 72 kDa) had two conserved catalytic motifs, an N-terminal Tryp_SPc domain (TS 15- 1n) and a C-terminal Tryp_SPc domain (TS 15-1c). To determine their collagen inducing ability, recombinant proteins (rTS 15-1n and rTS 15-1c) were produced using the pET-28a expression system. TS 15-1 is highly expressed during the muscle larval stage and has strong antigenicity. We determined that rTS 15-1c could elevate collagen I via activation of the TGF-β1 signaling pathwayin vitroandin vivo.Conclusion/SignificanceIn conclusion, we identified a host collagen inducing factor fromT. spiralisES-P using immunoscreening and demonstrated its molecular characteristics and functions.Author SummaryTrichinella spiraliscan make collagen capsules in host muscle cells during its life cycle, which encapsulates muscle stage larvae. Many investigators have tried to reveal the complex mechanism behind this collagen capsule architecture, and it has been suggested that several serine proteases in excretory-secretory proteins of the parasite are potential collagen capsule inducing factors. In addition, collagen synthesis is activated through the TGF-β/Smad signaling pathway and these events are closely related with protease activated receptor 2 which was activated by various serine proteases. In this study, we isolated and characterized a collagen gene expression inducer fromT. spiralisES-P using immunoscreening and investigated the candidate protein for its usefulness as a wound healing therapeutic agent.

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FDG PET/CT and N0 mediastinal nodal status in early stage non-small cell lung cancer: A single institution retrospective experience.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.11570 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. 11570-11570

Author(s):

Shashank Reddy Cingam ◽

Sean Connor ◽

Abhishek Harshad Patel ◽

Samip R. Master ◽

Amol Takalkar

Keyword(s):

Fdg Pet ◽

Early Stage ◽

False Negative ◽

Nodal Involvement ◽

True Negative ◽

Negative Results ◽

Early Stage Nsclc ◽

False Negative Results ◽

Pet Ct ◽

Fdg Pet Ct

11570 Background: The increasing use of CT and now low-dose screening CT scans for at-risk patients have led to increasing detection of lung cancer at early stages. FDG PET/CT is used as an adjunct to conventional imaging to assess loco-regional lymph node spread. However, there is a potential for false-negative results, especially in smaller lesions or early nodal involvement. The main objective of this study was to study the value of PET/CT scan to evaluate for true negative mediastinal nodes in patients with early stage NSCLC. Accurate determination of N0 status can have a significant impact on the cost-effectiveness and timely management of early stage NSCLC. Methods: Of a total of 404 patients with NSCLC managed at our facility between 2008 to 2015, 29 adult patients whose PET scan showed no or equivocal mediastinal nodal involvement and subsequently underwent surgical exploration of mediastinal lymph nodes were included in the study. Data variables that were collected included the cancer site, date of PET, node status on PET, type and date of surgery, cancer histology, and the tissue diagnosis of the sampled nodes. SAS software was used for the analysis of the data. Results: Of the 29 patients with N0 or equivocal nodes on FDG PET/CT, 7 (24.13%) had evidence of malignancy on biopsy of the surgically resected lymph nodes. No statistically significant differences were noted between the site of the neoplastic lesion, cancer histology, duration between the date of PET and the date of surgery in the true negative (TN) and false negative (FN) groups. The recurrence rate was higher in the FN group 60 % (3/5) compared to 21 % (4/19) in the TN group who had follow up for atleast 2 years. Conclusions: Our findings suggest incidence of false negative results of FDG PET/CT for evaluation of N0 nodes in early stage NSCLC is 24.13%. This is comparatively higher than false negatives rates with mediastinoscopy (5-10%) reported in other studies. Although our sample size is small, if confirmed, such a relatively high incidence of false negative results on FDG PET/CT for N0 disease supports the current recommendations for exploratory mediastinoscopy and/or surgery for definitive staging in early stage NSCLC.

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Identification of stage-specifically expressed genes of Trichinella spiralis by suppression subtractive hybridization

Parasitology ◽

10.1017/s0031182007002855 ◽

2007 ◽

Vol 134 (10) ◽

pp. 1443-1455 ◽

Cited By ~ 37

Author(s):

M. Y. LIU ◽

X. L. WANG ◽

B. Q. FU ◽

C. Y. LI ◽

X. P. WU ◽

...

Keyword(s):

Suppression Subtractive Hybridization ◽

Serine Proteases ◽

Zinc Finger Protein ◽

Trichinella Spiralis ◽

Gene Families ◽

Subtractive Hybridization ◽

Cdna Clones ◽

Dnase Ii ◽

Finger Protein ◽

Old Adult

SUMMARYNewborn larvae (NBL) and adult (Ad) stage-specifically expressed genes or members of gene families of Trichinella spiralis were identified by suppression subtractive hybridization (SSH)†. Six cDNA clones were identified as NBL stage-specific, including 1 member of the T. spiralis gene family encoding glutamic acid-rich proteins, 2 clones encoding novel serine proteases, 2 closely related clones encoding proteins that are members of a deoxyribonuclease II (DNase II)-like family and 1 clone with no similarity to known genes. Four stage-specific clones encoding homologues of retinoid X receptor, caveolin, C2H2 type zinc finger protein and a putative protein with no homology to known sequences were obtained from 3-day-old adult worms. One gene specifically up-regulated in the 5-day-old adult worms encoding a putative cuticle collagen was also identified.

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A Strain-Specific Antigen in Japanese Helicobacter pylori Recognized in Sera of Japanese Children

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.12.11.1280-1284.2005 ◽

2005 ◽

Vol 12 (11) ◽

pp. 1280-1284 ◽

Cited By ~ 9

Author(s):

Masumi Okuda ◽

Toshiro Sugiyama ◽

Kenichi Fukunaga ◽

Masaru Kondou ◽

Eikichi Miyashiro ◽

...

Keyword(s):

Molecular Weight ◽

Helicobacter Pylori ◽

High Molecular Weight ◽

Early Stage ◽

False Negative ◽

Specific Antigen ◽

Host Immune System ◽

Japanese Strain ◽

Asymptomatic Children ◽

Associated Proteins

ABSTRACT An enzyme immuno assay (EIA) test based on Japanese strain-derived high-molecular-weight cell-associated proteins (JHM-CAP) was evaluated by comparing with a previously developed EIA test based on a U.S. strain-derived high-molecular-weight cell-associated proteins (HM-CAP). Serum samples of 131 Japanese asymptomatic children (mean age, 5.5 years; range, 0 to 21 years) were tested that include 43 positive and 88 negative children as judged by Helicobacter pylori stool antigen test (HpSA test). Both tests showed comparable and reliable specificities, but the sensitivity of JHM-CAP EIA, at 93.0%, was much higher than that of HM-CAP EIA, at 67.4%. More false-negative results of HM-CAP were obtained in children under 10 years of age. Immunoblot analysis revealed that the JHM-CAP but not the HM-CAP preparation had a 100-kDa antigen recognized by JHM-CAP positive sera. It was concluded that JHM-CAP EIA is highly accurate for the serodiagnosis of H. pylori infection in Japanese young children and that the high sensitivity of JHM-CAP EIA in contrast to HM-CAP EIA is due to the presence of a 100-kDa antigen in Japanese strains that may be recognized by the host immune system at an early stage of infection.

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Ultrasonography in the study of hepatocellular carcinoma in woodchucks chronically infected with WHV

Laboratory Animals ◽

10.1258/002367703766453083 ◽

2003 ◽

Vol 37 (3) ◽

pp. 233-240 ◽

Cited By ~ 8

Author(s):

D. Lisi ◽

L. A. Kondili ◽

M. T. Ramieri ◽

R. Giuseppetti ◽

R. Bruni ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Hepatitis Virus ◽

Early Stage ◽

False Negative ◽

Normal Liver ◽

Liver Parenchyma ◽

Microscopical Examination ◽

Negative Results ◽

Chronic Hepatitis B Virus ◽

Us Findings

The woodchuck hepatitis virus (WHV)/woodchuck system is studied as animal model of human hepatocellular carcinoma (HCC) induced by chronic hepatitis B virus infection. The aim of the present study was the evaluation of ultrasound (US) liver examination in woodchuck as a routine method to detect HCC nodules and to follow their growth. Sixteen woodchucks were included in the study. US liver examination was carried out in all animals using a 5 MHz convex scanner. Macroscopic and microscopic examinations were performed to evaluate the US findings. The lower limit of nodule detection by US examination was a diameter of 5 mm. Macroscopic and microscopic examinations confirmed US findings in 14 of 16 animals (86.6%). No false negative results were obtained. Increase of nodule size was faster in the early phase of tumour growth. Small nodules (16 ± 5 mm) appeared as hypoechoic lesions with well-defined margins and homogeneous structure. Large nodules (42 ± 19 mm) appeared as hyperechoic lesions with irregular margins, heterogeneous or of mixed pattern; microscopical examination showed different degrees of necrosis, inflammation and fibrosis inside these latter neoplasms. The hepatitis reaction was conspicuously more severe around HCC nodules. No fibrosis and/or cirrhosis were found in normal liver parenchyma surrounding tumour nodules. On the whole, US appears to be helpful in the diagnosis of woodchuck HCC even at an early stage. Serial US evaluation can be used to study the growth rate of tumour nodules during natural history or experimental HCC treatments in woodchuck.

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Clinical Application of the Novel Cell-Based Biosensor for the Ultra-Rapid Detection of the SARS-CoV-2 S1 Spike Protein Antigen: A Practical Approach

Biosensors ◽

10.3390/bios11070224 ◽

2021 ◽

Vol 11 (7) ◽

pp. 224

Author(s):

Sophie Mavrikou ◽

Vasileios Tsekouras ◽

Kyriaki Hatziagapiou ◽

Foteini Paradeisi ◽

Petros Bakakos ◽

...

Keyword(s):

Mammalian Cells ◽

Early Stage ◽

False Negative ◽

Sample Pretreatment ◽

High Sensitivity ◽

Spike Protein ◽

The Novel ◽

Major Step ◽

Negative Results ◽

Potential Applicability

The availability of antigen tests for SARS-CoV-2 represents a major step for the mass surveillance of the incidence of infection, especially regarding COVID-19 asymptomatic and/or early-stage patients. Recently, we reported the development of a Bioelectric Recognition Assay-based biosensor able to detect the SARS-CoV-2 S1 spike protein expressed on the surface of the virus in just three minutes, with high sensitivity and selectivity. The working principle was established by measuring the change of the electric potential of membrane-engineered mammalian cells bearing the human chimeric spike S1 antibody after attachment of the respective viral protein. In the present study, we applied the novel biosensor to patient-derived nasopharyngeal samples in a clinical set-up, with absolutely no sample pretreatment. More importantly, membrane-engineered cells were pre-immobilized in a proprietary biomatrix, thus enabling their long-term preservation prior to use as well as significantly increasing their ease-of-handle as test consumables. The plug-and-apply novel biosensor was able to detect the virus in positive samples with a 92.8% success rate compared to RT-PCR. No false negative results were recorded. These findings demonstrate the potential applicability of the biosensor for the early, routine mass screening of SARS-CoV-2 on a scale not yet realized.

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Contrast-Enhanced MR Imaging of the Breast in Patients with Breast Implants after Cancer Surgery

Acta Radiologica ◽

10.1177/028418519503600201 ◽

1995 ◽

Vol 36 (2) ◽

pp. 111-116 ◽

Cited By ~ 15

Author(s):

B. Boné ◽

P. Aspelin ◽

B. Isberg ◽

L. Perbeck ◽

B. Veress

Keyword(s):

Mr Imaging ◽

Early Stage ◽

False Negative ◽

Breast Implants ◽

Breast Cancer Patients ◽

Negative Results ◽

Contrast Enhanced ◽

Investigation Methods ◽

Size Type ◽

After Cancer

The purpose of the study was to determine the value of contrast-enhanced MR imaging in the assessment of local recurrence in breast cancer patients who underwent mastectomy and breast reconstruction with an implant. Eighty-three patients have been evaluated by semidynamic contrast-enhanced MR imaging. The T1-weighted FLASH 3-D sequence was repeated twice postcontrast for evaluation of the entire breast bilaterally. The findings were compared to physical examination, mammography and histopathology. Recurrence verified by histopathology occurred in 14 of 83 patients (17%). Contrast-enhanced MR imaging was superior to palpation and mammography in revealing recurrences, especially when these were located close to the chest wall. MR was also more sensitive in detecting multiple foci of cancers. Our study revealed that MR imaging was influenced by size, type and composition of the tumor, as illustrated by the false-negative results. Therefore, the use of all 3 investigation methods is necessary for detecting recurrence at an early stage during the postoperative follow-up.

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Nucleic Acid and Immunological Diagnostics for SARS-CoV-2: Processes, Platforms and Pitfalls

Diagnostics ◽

10.3390/diagnostics10110866 ◽

2020 ◽

Vol 10 (11) ◽

pp. 866

Author(s):

Avinash Premraj ◽

Abi George Aleyas ◽

Binita Nautiyal ◽

Thaha J Rasool

Keyword(s):

Sensitivity And Specificity ◽

Early Stage ◽

Control Strategies ◽

False Negative ◽

Primary Diagnosis ◽

Test Methods ◽

Molecular Technique ◽

Negative Results ◽

False Negative Results ◽

Host Tissues

Accurate diagnosis at an early stage of infection is essential for the successful management of any contagious disease. The coronavirus disease 2019 (COVID-19), caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) virus is a pandemic that has affected 214 countries affecting more than 37.4 million people causing 1.07 million deaths as of the second week of October 2020. The primary diagnosis of the infection is done either by the molecular technique of RT-qPCR by detecting portions of the RNA of the viral genome or through immunodiagnostic tests by detecting the viral proteins or the antibodies produced by the host. As the demand for the test increased rapidly many naive manufacturers entered the market with novel kits and more and more laboratories also entered the diagnostic arena making the test result more error-prone. There are serious debates globally and regionally on the sensitivity and specificity of these tests and about the overall accuracy and reliability of the tests for decision making on control strategies. The significance of the test is also complexed by the presence of asymptomatic carriers, re-occurrence of infection in cured patients as well as by the varied incubation periods of the infection and shifting of the viral location in the host tissues. In this paper, we review the techniques available for SARS-CoV-2 diagnosis and probable factors that can reduce the sensitivity and specificity of the different test methods currently in vogue. We also provide a checklist of factors to be considered to avoid fallacious practices to reduce false positive and false negative results by the clinical laboratories.

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Does aggressive management of solitary pulmonary nodules pay off?

Breathe ◽

10.1183/20734735.0275-2018 ◽

2019 ◽

Vol 15 (1) ◽

pp. 15-23 ◽

Cited By ~ 4

Author(s):

Stefano Elia ◽

Serafina Loprete ◽

Alessandro De Stefano ◽

Georgia Hardavella

Keyword(s):

Lung Cancer ◽

Minimally Invasive ◽

Early Stage ◽

Large Diameter ◽

False Negative ◽

Pulmonary Nodules ◽

Solitary Pulmonary Nodules ◽

Early Stage Lung Cancer ◽

Fdg Uptake ◽

Negative Results

Indeterminate solitary pulmonary nodules (SPNs), measuring up to 3 cm in diameter, are incidental radiological findings. The ever-growing use of modern imaging has increased their detection. The majority of those nodules are benign; however, the possibility of diagnosing early-stage lung cancer still stands. Guidelines for the management of SPNs have never been validated in prospective comparative studies.Positron emission tomography (PET) is a useful tool to provide functional information on SPNs. However, overall sensitivity and specificity of PET in detecting malignant SPNs of at least 10 mm in diameter are about 90% and false-negative results are reported.The development of video-assisted thoracic surgery has provided minimally invasive diagnosis and treatment of SPNs. In our series, 105 patients underwent surgery based on combined increased18F-labelled 2-fluoro-2-deoxy-d-glucose (FDG) uptake on PET computed tomography and radiological features (morphology and density) without prior histological confirmation. We detected 26 false negatives (24.8%) and only nine false positives (8.57%). Therefore, our minimally invasive surgical approach prevented 25% of patients with lung cancer from a delayed treatmentversusonly 9% undergoing “overtreatment”.In our monocentric cohort, patients with SPNs with large diameter, irregular outline, no calcifications, central location, increased FDG uptake and/or subsolid aspect benefited from a primary surgical resection.

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