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Imaging of objects through active electrolocation in Gnathonemus petersii.

The weakly electric fish Gnathonemus petersii detects, localizes, and analyzes objects during active electrolocation even in complete darkness. This enables these fish to lead a nocturnal life and find and identify their prey (small insect larvae) on the ground of their freshwater habitat. During active electrolocation, fish produce a series of brief electric signals, electric organ discharges (EOD), with an electric organ in their tail. Each EOD builds up a stable electric field around the fish, which is distorted only by nearby objects. Field distortions lead to changes of the transepidermal electric current flow at a region of the fish's electroreceptive skin surface called the 'electric image'. Within the electric image, locally perceived EODs can be either altered in amplitude or waveform by an object. Fish measure both parameters to assess object properties, such as the capacitive and resistive components of the object's complex impedance. the object's size and shape, and its distance from the fish. None of these object properties can be evaluated in isolation, but have to be inferred during parallel processing of electric image spatial and qualitative parameters. Two anterior skin regions of G. petersii appear to possess particular properties for special electrolocation tasks and we therefore refer to them as 'foveal' regions. Because of its high electroreceptor density, the electric field geometry around it, and its behavioral use, the 'nasal region' between the nares and the mouth at the head of the fish is suggested to be a fovea for long-range guidance and object detection. We propose that the 'Schnauzenorgan', a long and flexible chin appendix covered densely with electroreceptor organs, is a second electroreceptive fovea associated with a short-range (food) identification system. Together, these two electric foveae constitute an effective prey detection and identification system.

Animals↗

The role of whole-body bone scanning and clinical factors in detecting bone metastases in patients with non-small cell lung cancer.

STUDY OBJECTIVES: Correct detection of bone metastases in patients with non-small cell lung cancer (NSCLC) is crucial for prognosis and selection of an appropriate treatment regimen. The aim of this study was to investigate the role of whole-body bone scanning (WBBS) and clinical factors in detecting bone metastases in NSCLC. DESIGN AND PATIENTS: One hundred twenty-five patients with a diagnosis made between 1998 and 2002 were recruited (squamous cell carcinoma, 54.4%; adenocarcinoma, 32.8%; non-small cell carcinoma, 8.8%; large cell carcinoma, 4%). Clinical factors suggesting bone metastasis (skeletal pain, elevated alkaline phosphatase, hypercalcemia) were evaluated. WBBS was performed in all patients, and additional MRI was ordered in 10 patients because of discordance between clinical factors and WBBS findings. MEASUREMENTS AND RESULTS: Bone metastases were detected in 53% (n = 21) of 39 clinical factor-positive patients, 5.8% (n = 5) of 86 clinical factor-negative patients, and 20.8% of total patients. The existence of bone-specific clinical factors as indicators of metastasis presented 53.8% positive predictive value (PPV), 94.2% negative predictive value (NPV), and 81.6% accuracy. However, the findings of WBBS showed 73.5% PPV, 97.8% NPV, and 91.2% accuracy. Adenocarcinoma was the most common cell type found in patients with bone metastasis (39%). The routine bone scanning prevented two futile thoracotomies (8%) in 25 patients with apparently operable lung cancer. CONCLUSIONS: In spite of the high NPV of the bone-specific clinical factors and the high value obtained in the false-positive findings in the bone scan, the present study indicates that in patients for whom surgical therapy is an option, preoperative staging using WBBS can be helpful to avoid misstaging due to asymptomatic bone metastases.

Adult↗

[Detection of circulating prostatic cells with RT-PCR PSA in prostatic cancer].

OBJECTIVES: The detection of circulating prostatic cells by molecular biology techniques (RT-PCR) can be useful in the staging of localized prostate cancer prior to radical prostatectomy in some institutions. After describing their technique, the authors report their results. PATIENTS: 80 RT-PCR were performed: 32 in a control group (including 11 women free of any neoplastic disease, 11 healthy men, and 10 men with benign prostatic hyperplasia before resection), and 48 in patients with prostate cancer (43 with clinically localized cancer and 5 with metastatic cancer). RESULTS: In the control group, none of 11 women had a positive RT-PCR, 1 of the healthy men was positive (orchidopexy) and 3 of the 11 patients with benign prostatic hyperplasia were positive, but none of them had tumour on the resection chips. None of the 5 metastatic patients were positive. In the patients treated by radical prostatectomy, no correlation was observed between RT-PCR results, pathological stage, positive resection margin status and laboratory progression after radical prostatectomy. CONCLUSION: This PSA RT-PCR technique developed in this institution does not appear to be useful for the molecular staging of prostate cancer. This study demonstrates the difficulty of standardization of this technique which limits its routine use.

Aged↗

[Detection of tumor cells in peripheral blood for patients with ovarian epithelial carcinoma].

OBJECTIVE: To detect tumor cell contamination of peripheral blood stem cell (PBSC) and the presence of tumor cells in peripheral circulating blood in patients with primary advanced ovarian cancer. METHODS: To establish limits of sensitivity of the immunocytochemical stainings technique (ABC method) for tumor cell detection, four types of ovarian cancer tumor cell(ovarian cancer cell lines SKOV3, 3AO, primary cancer cell from ascites of serous papillary carcinoma and undifferentiated adenocarcinoma) were added into normal blood to prepare the models with different ratios of tumor cells to mononuclear cells. Monoclonal antibodies(MAB) of COC 183 B2, CK(AE1/AE3) and their cocktail antibody (mixture of them) were used for immunocytochemical stainings. 14 blood specimens were examined (6 stem cell blood specimens, 8 primary advanced ovarian cancer blood spcimens). RESULTS: The sensitivity of detection achieved by cocktail antibodies was superior to either COC183B2 or CK MAb alone. The sensitivity of cocktail antibody was 1:5 x 10(5). According to the established sensitivity, none of the fourteen specimens contained tumor cell. CONCLUSIONS: None of the 6 PBSC specimens obtained from 2 patients and eight peripheral blood specimens obtained from 8 patients with primary ovarian epithelial carcinoma contained tumor cell.

Adenocarcinoma↗

[Detection of circulating pemphigus vulgaris antibodies in families of patients with pemphigus vulgaris].

OBJECTIVE: To detect the autoantibodies on 30 first-degree relatives of 12 patients with pemphigus vulgaris (PV), who may share the same susceptible gene with the patients. METHODS: Immunoblot and indirect immunofluorescence (IIF) method were used to detect circulatory PV antibodies in the peripheral blood. RESULTS: PV antibodies were found in 19 of 30 (63%) relatives by both methods, but 10 control serum were all negative by both methods. CONCLUSION: Healthy PV antibody carriers may also exist in high frequency in the first-degree relatives of Chinese PV patients.

Adolescent↗

[Detection of Coxsackie B virus IgM antibody in myocarditis patients by Mac ELISA].

OBJECTIVE: To detect Coxsackie B virus type 1-6 IgM antibodies from 193 clinically diagnosed myocarditis cases. METHODS: The capture IgM ELISA (Mac ELISA) test was used. RESULTS: The total positive rate of Coxsackie virus B 1-6 IgM antibody in myocarditis patients was 69.6% (135 cases), among which Cox B3 type was as high as 40% (54 cases) of the total positive patients. The others were 29.6% (40 cases), 18.5% (25 cases), 7.4%(10 cases), 3.0% (4 cases), 1.5% (2 cases) of Cox B2, Cox B4, Cox B1, Cox B5 and Cox B6 respectively. Meanwhile only 20.4% (37 cases) Cox B IgM positive cases could be detected in 181 non-myocarditis patients of same ages. The results showed that the positive rate of Cox B virus IgM antibody in clinically diagnosed myocarditis was significantly higher than that in control group (P <0.01). CONCLUSIONS: The Cox B viruses may be the main agents causing viral myocarditis in Beijing cases. Mac ELISA is an easy, rapid, specific and sensitive method to detect Cox B virus IgM antibody for early and rapid diagnosis of myocarditis.

Antibodies, Viral↗

mRNA of MAGE genes as specific markers in detection of tumor cells in the peripheral blood of patients with hepatocellular carcinoma.

OBJECTIVE: To detect tumor cells in the peripheral blood of patients with hepatocellular carcinoma (HCC) by using the mRNA of the MAGE-1 and MAGE-3 genes as specific tumor markers. METHODS: Peripheral blood was obtained from 25 HCC patients and 20 healthy volunteers. The mRNA of the MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMCs) was detected by nested RT-PCR. The MAGE-1 and MAGE-3 transcripts in the tumor tissues of these HCC patients were also detected by RT-PCR. RESULTS: Of the 25 HCC patients, MAGE-1 and MAGE-3 mRNA were positive in 44% (11/25) and 36% (9/25) of PBMCs respectively, and in 68% (17/25) and 56% (14/25) of HCC tissues respectively. In the PBMCs of the 25 HCC patients, 16 (64%) samples were detected to express at least one type of MAGE mRNA. MAGE mRNA were not detected in the PBMCs from the patients whose tumors did not express the MAGE genes, nor in the PBMCs from the 20 healthy donors. The positive rate of MAGE mRNA in the PBMCs was closely correlated with the TNM stages and the diameter of tumors, but there was no correlation between the positive rate of MAGE mRNA in PBMCs and tumor differentiation degree or serum alpha-FP level. Of 9 HCC patients whose serum alpha-FP was normal or slightly elevated (< 50 ng/ml), 6 were MAGE-1 and/or MAGE-3 mRNA positive in their PBMCs. CONCLUSION: MAGE-1 and MAGE-3 mRNA could be specifically detected with high percentage in the PBMCs of HCC patients by our method. They can be used as specific tumor markers for the detection of the circulating HCC cells, and the detection results may be helpful to evaluate the prognosis of HCC patients.

Antigens, Neoplasm↗

[Circulating antigen detection in schistosomiasis japonica patients by sandwich ELISA using polyclonal and monoclonal antibodies].

OBJECTIVE: To detect circulating antigen in the patients with schistosomiasis. METHODS: Polyclonal antibody(PcAb)-monoclonal antibody(McAb) sandwich ELISA. RESULTS: Sera from 150 patients with schistosomiasis were detected, the average sensitivity was 84.7%(72.0%-91.0%). No false positive reaction was detected in 40 normal controls and none but 2 patients(2/10) with paragonimiasis showed cross reactions among 74 cases with other parasitic infections. CONCLUSION: PcAb-McAb sandwich ELISA is a relatively ideal method for circulatling antigen detection.

Animals↗

[Detection of chromosome 8 anomalies in ovarian carcinoma by fluorescence in situ hybridization].

OBJECTIVE: To detect the relationship between chromosomal anomalies and the pathogenesis, development and prognosis of ovarian carcinoma. METHODS: Thirty-six specimens of ovarian carcinoma (n=12), ovarian benign tumor (n=12), and normal ovary (n=12) were examined by fluorescence in situ hybridization (FISH). RESULTS: Twelve cases of mutations, including trisomy 8, monosomy 8 or tetraploid 8 chromosomal anomalies, were found in the group of ovarian carcinoma, making up 100% (12/12). Three cases of trisomy 8 chromosomal anomalies were found in the group of ovarian benign tumor, accounting for 25% (3/12). No anomaly was found in the normal group. There were significant differences between the three groups, P<0.001. CONCLUSION: The above anomalies of chromosome 8 are significantly associated with the pathogenesis and development of ovarian carcinoma. The anomalies may occur in the early stage of the carcinoma, and may be significantly associated with the pathological differentiation and clinical stage of the case.

Adolescent↗

[Mutation detection of PKD2 gene in Chinese by denaturing high-performance liquid chromatograph].

OBJECTIVE: To detect the mutations of autosomal dominant polycystic kidney disease gene 2(PKD2)in Chinese. METHODS: The white blood cell genomic DNA from patients of 94 Chinese autosomal dominant polycystic kidney disease(ADPKD) pedigrees was isolated and amplified by polymerase chain reaction(PCR). The PCR products were analyzed by denaturing high-performance liquid chromatography(DHPLC). The samples with abnormal profiles were sequenced. RESULTS: Eight mutations were identified, including 2 nonsense mutations, 2 deletion mutations,1 insertion mutation and 3 missense mutations. Two nonsense mutations occurred in exon 5(1249C-->T) and exon 13(2407C-->T),both resulted in a stop codon. The insertion was in exon 2(636-637 ins T),and the deletion mutations were in exons 12(2348-2351 del AGAA) and 13(2401 delete A),resulting in the reading frame shift. Three missense mutations were in exons 1(G568-->A),4(C964-->T),and 5(G1168-->A), which caused amino acid changes (190Ala-->Thr,322Arg-->Trp,390Gly-->Ser). CONCLUSION: The method of DHPLC was used in detecting mutations successfully and 8 mutations in PKD2 were identified. It will be useful in the molecular diagnosis of ADPKD in advance of the cysts formation and birth.

Adolescent↗

[Detection of hypermethylation of p16 gene in plasma DNA from lung cancer patients].

OBJECTIVE: To detect hyper methylation of p16 gene in plasma DNA from patients with lung cancer, and to assess its potential as a malignant marker. METHODS: Using a modified semi-nested methylation-specific PCR (MSP), the status of methylation of the p16 was investigated in plasma DNA from 137 lung cancer patients and 112 matched tumor tissues. RESULTS: Hypermethylation of the p16 was present in 75.2% (103/137) of the plasma samples and 80.4% (90/112) of the tumor tissues. Hypermethylation of the p16 in the plasma was detected in 77.9% squamous-cell carcinoma, 65.1% adenocarcionma, 75.1% adeno-squamous-cell carcinoma, and 91.7% small-cell lung cancer. Only in those patients whose tumor tissues had hypermethylation of p16 gene, similar changes could be detected in their plasma samples. Hypermethylation of the p16 in plasma and the corresponding tumor tissues was not significantly correlated with the clinical stage and pathological type of the tumor. CONCLUSION: The result indicates that hypermethylation of the p16 may be a useful marker in the auxiliary diagnosis of lung cancer.

Adenocarcinoma↗

[Detection of HPV 58 and cloning of its E7 gene in cervical cancer].

OBJECTIVE: To detect HPV 58, a common type of human papillomavirus (HPV), clone and express its E7 gene from biopsy specimens of cervical cancer. METHODS: HPV 58 from 58 biopsy tissues of cervical cancer was detected by GP5+/GP6+ PCR followed by template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP). HPV 58 E7 gene was amplified from one HPV 58-positive sample, and then cloned into pGEM-T Easy vector. The recombinant plasmid, HPV58E7-pGEM-T was confirmed by sequencing. Subsequently, E7 gene was cloned into prokaryotic expression vector pRSET-A. The constructed pRSET-58E7 plasmids were transfected into BL21(DE3) cells, and induced to express 58 E7 protein by IPTG. RESULTS: Among the 58 biopsy tissues of cervical cancer, 10 were HPV 58-positive, accounting for 19.2% of 52 HPV-positive cases. HPV 58 E7 gene was amplified from one HPV 58-positive sample. The constructed plasmids were identified containing HPV58 E7 gene by restriction enzyme analysis and sequencing. SDS-PAGE analysis showed that HPV58 E7 His6 fusion protein of M(r) 16 x 10(3) was expressed by pRSET-58E7 after induction by IPTG. The fusion protein accounted for 30% of total bacterial proteins. CONCLUSION: HPV 58 is not uncommon in Chinese women with cervical cancer in Shaanxi province. Constructed HPV58 E7 recombinant plasmids can be effectively expressed in E.coli, which may provide a tool in diagnosis and vaccine design for HPV of HPV58-associated tumors.

Adult↗

[Detection of fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia].

OBJECTIVE: To detect the expression of the fusion genes resulting from chromosome abnormalities in childhood acute lymphoblastic leukemia(ALL) and its conformity to WHO classification. METHODS: Sixty-two children with ALL were investigated. The expression of fusion genes was determined by multiplex reverse transcription-polymerase chain reaction (RT-PCR), karyotyping (R band) and immunophenotyping (by flow cytometry) were also performed. RESULTS: Of the 62 patients, 23(37.1%) were found to carry 13 different fusion genes. The patients with immunophenotype of Pre-B-ALL were found to carry: TEL/AML1(3 cases); E2A/PBX1, E2A/HLF, TLS/ERG, MLL/AF4, MLL/AF9, MLL/AF10, MLL/AFX-MLL/AF6-MLL/ELL, MLL/AF6-MLL/ELL, dupMLL (one case for each); and HOX11 (6 cases). The patients with immunophenotype of Pre-T-ALL were found to carry: TAL1D (4 cases, one is also found to have HOX11 expression); and HOX11 (2 cases). The multiplex RT-PCR in combination with chromosome analysis revealed genetic abnormalities in 69.4%(43/62) of childhood ALL. CONCLUSION: Multiplex RT-PCR combined with chromosome analysis and immunophenotyping can provide reliable and helpful information for the diagnosis, therapy evaluation and prognosis prediction in childhood ALL, which may also serve as a basis on which to implement the criteria of WHO classification.

Adolescent↗

[Detection of circulating hypermethylated tumor-specific RASSF1A DNA in ovarian cancer patients].

OBJECTIVE: To detect hypermethylated tumor-specific RASSF1A DNA in the circulation and its significance in ovarian cancers patients. METHODS: Methylation-specific polymerase chain reaction (MSP) was used to study the hypermethylation of RASSF1A in preoperative serum samples from 51 ovarian cancer patients. RESULTS: The RASSF1A gene was not methylated in peripheral blood samples from 51 normal patients and 51 patients with benign ovarian tumors. Hypermethylation of RASSF1A gene was found in circulating tumor-specific DNA in 43.1% of patients (22 out of 51 cases) with ovarian cancers (P < 0.05). There was no difference in hypermethylation of RASSF1A gene amongst various ovarian cancer subtypes (P < 0.05). On the other hand, hypermethylation of RASSF1A gene was more frequently encountered in stage III and IV than stage I and II tumors (P < 0.05). It was rarely seen in well and moderately differentiated groups, as compared with poorly differentiated group (P < 0.05). CONCLUSIONS: There is a higher frequency of RASSF1A hypermethylation in circulating tumor-specific DNA of ovarian cancer patients. RASSF1A has been postulated to play an important role as tumor suppressor gene and can be silenced by promoter hypermethylation. This methylation correlates with clinical stage and histopathologic grade. Such observation may carry diagnostic and prognostic implications when assessing ovarian tumors.

Carcinoma, Endometrioid↗

Pneumocystis jiroveci in HIV/AIDS patients: detection by FTA filter paper together with PCR in noninvasive induced sputum specimens.

OBJECTIVES: To detect P. jiroveci (previously named P. carinii) by PCR using FTA filter paper to extract the DNA, from noninvasive induced sputum samples of HIV/AIDS patients. MATERIAL AND METHOD: Fifty two HIV/AIDS patients suspected of Pneumocystis jiroveci pneumonia (PJP) in King Chulalongkorn Memorial Hospital were recruited. Both cytological method and PCR with FTA filter paper technique were performed to detect P jiroveci from each specimen. RESULTS: The detectability rate of P. jiroveci infection was 21%. The PCR with FTA filter paper method was 4 folds much more sensitive than Giemsa staining technique. P. jiroveci was detected in 18% of the HIV/AIDS patients in spite of receiving standard PJP prophylaxis. CONCLUSION: Detection of P. jiroveci by using FTA filter paper together with PCR in induced sputum samples could detect more cases of P. jiroveci infection than by using cytological method. DNA extraction using the FTA filter paper was more rapid and convenient than other extraction methods. The causes of failure of PJP prophylaxis should be evaluated.

AIDS-Related Opportunistic Infections↗

[Value of lymph node micrometastasis detected by RT-PCR assay in determining the stage of gastric carcinoma].

OBJECTIVE: To detect micrometastasis in regional lymph nodes using RT- PCR assay and evaluate the significance of the new assessment of nodal status in determining pN staging in gastric carcinoma. METHODS: In addition to HE,RT- PCR assay for cytokeratin-20 gene marker was used to detect micrometastasis in 850 lymph nodes from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenectomy from Dec. 2003 to Apr. 2004. RESULTS: The detection rate of HE staining was 27.1% (233/850), the detection rate of RT-PCR assay was 36.5% (310/850) (Pa< 0.01). Lymph node micrometastasis was further detected in 77 nodes from 14 patients. The detection rate of RT- PCR assay from the routine negative lymph nodes was 12.5% (77/617). Seven of those patients were up- staged (from IB stage to II stage, from IB stage to III A stage, from I stage to III A stage, from III A stage to III B stage, from III A stage to IV stage in one patient respectively, and from III B stage to IV stage in 2 patients). CONCLUSION: RT- PCR assay can increase the detection rate of lymph node metastasis and have a significant impact on the staging system of gastric carcinoma.

Adult↗

Prenatal detection of heart defects in a non-selected population of 30,149 fetuses--detection rates and outcome.

OBJECTIVES: To evaluate the detection rate of congenital heart defects (CHD) in a non-selected population and to follow outcome after diagnosis. METHODS: All 30,149 fetuses/newborns that were scheduled to deliver at our hospital between February 1991 and December 2001 were registered prospectively. Of these, 29,460 (98%) fetuses had a prenatal ultrasound scan at our center. The routine fetal examination at approximately 18 weeks' gestation included the four-chamber view and the great arteries of the fetal heart. The follow-up period was 2-13 years. RESULTS: Of 97 major CHDs, 55 (57%) were detected prenatally, 16% (9/55) prior to, 66% (36/55) at and 18% (10/55) after the routine scan. Forty-four percent (19/43) of the isolated CHDs, 67% (36/54) of those with associated malformations and 48% (11/23) of the isolated ductal-dependent CHDs were detected. Thirty-eight percent (37/97) had an abnormal karyotype. Of the 55 major CHDs detected, 44% (24) of the pregnancies with lethal/serious fetal malformations were terminated, 15% (8) died in utero, 42% (23) were born alive and 27% (15) were still alive after 2 years. Of the 42 CHDs detected postnatally, 2% (1) were terminated for other reasons, 98% (41) were born alive and 81% (34) were still alive after 2 years. CONCLUSIONS: Prenatal detection of CHD is still a challenge, with a 57% detection rate only. Isolated defects are detected less frequently. The overall outcome suggests that the most severe defects are detected with the present screening setting; only 27% of the babies with major CHDs detected were still alive after 2 years. Data from long-term follow-up will be of importance for the counseling process.

Adolescent↗

Pregnancy outcome after early detection of bacterial vaginosis.

OBJECTIVE: To assess if detecting bacterial vaginosis either in early pregnancy or at midtrimester may predict adverse pregnancy outcome in women at risk for preterm delivery. STUDY DESIGN: 242 pregnant women with a previous preterm delivery were evaluated for bacterial vaginosis either in the first trimester (prior to 10+0 weeks) or in the second one (24-26 weeks). Adverse outcome was intended as miscarriage (< or =25 weeks), or premature delivery (< or =36+6). RESULTS: The risk of adverse pregnancy outcome was significantly increased in women diagnosed at first trimester with bacterial vaginosis (OR: 4.56; 95% CI: 2.54-8.93); the same finding at midtrimester did not increase significantly the risk of preterm delivery. CONCLUSIONS: Early screening for bacterial vaginosis in pregnant women who experienced a preterm delivery may help in predicting the risk of adverse outcome.

Abortion, Spontaneous↗