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Biomedical subjects

Y M Lo

Publications and source records attributed to Y M Lo.

At least 37 records · Page 2Linked to original sources
PubMed2000

Prenatal detection of fetal Down's syndrome from maternal plasma.

Fetal DNA is present in maternal plasma, and a proportion of such DNA is seen in intact fetal cells. We investigated the use of fluorescence in-situ hybridisation (FISH) techniques on maternal plasma samples. In plasma samples obtained from three women carrying fetuses affected by trisomy 21 (Down's syndrome), we identified fetal cells with three chromosome-21 signals. These results show the feasibility of non-invasive detection of fetal chromosomal aneuploidy by maternal plasma analysis.

Down Syndrome↗
PubMed2000

Kinetics of plasma Epstein-Barr virus DNA during radiation therapy for nasopharyngeal carcinoma.

We studied the kinetics of circulating EBV DNA in the plasma of nasopharyngeal carcinoma (NPC) patients. Serial weekly sampling of 10 NPC patients revealed a rapid decline in plasma EBV DNA concentration after treatment. In two subjects, an initial rise in the circulating EBV DNA level was observed immediately after treatment initiation. Plasma EBV DNA levels were monitored daily during the first treatment week in a second cohort of five patients, and the results indicated that an initial rise in plasma EBV DNA concentration could be observed in all subjects during the first treatment week. This observation is consistent with the liberation of EBV DNA after therapy-induced cancer cell death. After this initial rise, plasma EBV DNA concentration was found to decay with a median half-life of 3.8 days (interquartile range, 2.4-4.4 days). Kinetic analysis of circulating tumor-derived DNA during treatment may be a powerful tool for evaluating the in vivo response of NPC and other tumors to antineoplastic treatment and may improve our understanding of the biology of plasma nucleic acids.

Carcinoma, Squamous Cell↗
PubMed2000

Quantitative analysis of circulating cell-free Epstein-Barr virus (EBV) DNA levels in patients with EBV-associated lymphoid malignancies.

Cell-free Epstein-Barr virus (EBV) DNA has recently been detected in the plasma and serum of patients with Hodgkin's disease, post-transplant lymphoproliferative disease (PTLD) and acquired immunodeficiency syndrome-related lymphoma. However, no data are available on the temporal variation of plasma/serum EBV DNA levels in patients with EBV-associated lymphoid malignancies during the course of therapy. Using a real-time quantitative polymerase chain reaction assay, we studied the plasma EBV DNA levels in 13 patients with EBV-associated lymphoid malignancies (six patients with Hodgkin's disease, four with nasal natural killer/T-cell lymphoma, two cases of PTLD and one patient with Burkitt's lymphoma) at presentation and during therapy. Plasma EBV DNA was detected in 12 of the 13 patients (median 2,266 copies/ml; interquartile range 181-8,379 copies/ml), but not in any of 35 healthy control subjects (P < 0.0001). The EBV status in tumour cells was also examined in 12 of these patients using in situ hybridization for EBV-encoded small RNAs (EBERs). EBER positivity was observed in 11 patients, all of whom had EBV DNA detectable in plasma. The one patient who had no detectable plasma EBV DNA was also negative for EBERs in tumour tissue. Serial measurements of plasma EBV DNA levels were performed in nine of the patients during the course of therapy. All patients who responded to therapy demonstrated a significant reduction of plasma EBV DNA to low or undetectable levels, whereas in two patients with ineffective therapy, disease progression was associated with a rapid increase in plasma EBV DNA levels. We concluded that plasma EBV DNA is detectable in a wide range of EBV-associated lymphoid malignancies. As plasma EBV DNA levels correlate well with the therapeutic response, such analysis may be a valuable tool for monitoring clinical progress.

Adult↗
PubMed2000

Cell-free fetal deoxyribonucleic acid in maternal circulation as a marker of fetal-maternal hemorrhage in patients undergoing external cephalic version near term.

OBJECTIVE: Our aim was to investigate whether external cephalic version performed near term increases the concentration of cell-free fetal deoxyribonucleic acid in maternal plasma. STUDY DESIGN: Forty-five patients who had singleton male fetuses and were undergoing external cephalic version at or beyond 36 weeks of gestation were recruited during a 20-month period. Maternal venous blood samples were taken before and within 10 minutes after external cephalic version. Deoxyribonucleic acid was extracted from the plasma samples. The amount of fetal deoxyribonucleic acid was quantified by means of the SRY gene on the Y chromosome as a fetal marker. The change in SRY gene concentration before and after external cephalic version was compared by paired sample t test. RESULTS: There was a significant increase in the concentration of fetal deoxyribonucleic acid in maternal serum after external cephalic version (before, 296 +/- 209 copies per milliliter; after, 369 +/- 228 copies per milliliter; P =.014). This increase in the concentration of deoxyribonucleic acid was most profound among the nulliparous patients after a successful version and in the presence of a posterior placenta. The location of the placenta was found to be the most significant factor accounting for the change in the deoxyribonucleic acid concentration. CONCLUSIONS: External cephalic version near term imposed a significant disturbance to the maternalplacental interface. Fetal deoxyribonucleic acid is a sensitive marker that is useful in the assessment of subclinical fetal-maternal hemorrhage.

Adult↗
PubMed2000

Fetal DNA in maternal plasma.

Recently, cell-free fetal DNA has been found in maternal plasma and serum. This discovery opens up a new field of investigation and provides an easily accessible source of fetal genetic material for prenatal diagnosis. Prenatal diagnostic applications of fetal DNA in maternal plasma include the investigation of sex-linked disorders and fetal rhesus D status determination. Cell-free fetal DNA has been found to be present in much higher fractional concentrations than fetal nucleated cells in maternal blood. The concentration of fetal DNA increases throughout pregnancy, with a sharp rise towards the end of gestation. Abnormally high levels of cell-free DNA have been found in pregnancies complicated by preeclampsia and preterm labor, an observation that has potential diagnostic and pathophysiologic implications. Much remains to be learned regarding the mechanisms of production and clearance of maternal plasma fetal DNA. It is hoped that the eagerly awaited answers to these and other questions may ultimately enhance our understanding of the fetomaternal relationship.

Cell-Free System↗
PubMed2000

Presence of mitochondrial tRNA(Leu(UUR)) A to G 3243 mutation in DNA extracted from serum and plasma of patients with type 2 diabetes mellitus.

AIMS/BACKGROUND: An A to G substitution at base pair 3243 in the mitochondrial tRNA(Leu(UUR)) gene (mt3243) is commonly associated with maternally inherited diabetes and deafness, and other diseases. It is possible that cell free mitochondrial DNA exists in serum and plasma from these patients, and these samples might be a source of material for the detection of such mutations. METHODS: Sixteen patients with type 2 diabetes mellitus and 25 healthy subjects were tested for the 3243 mutation by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. Plasma and serum from the 41 subjects were tested blind, without knowledge of the final diagnosis. RESULTS: PCR amplification of the mtRNA(Leu(UUR)) region in mitochondrial DNA (mtDNA) in serum samples revealed the presence of mtDNA in all samples. After ApaI digestion of the amplified DNA fragments, mt3243 was detected in the serum and plasma samples of the seven patients with diabetes who had previously been found to have this mutation in their leucocyte DNA. None of the serum/plasma samples from the healthy subjects or those patients negative for mt3243 in their leucocytes had this mutation (p < 0.001). In addition, the degree of heteroplasmy of mt3243 appeared to be higher in serum and plasma samples than in leucocytes among mt3243 carriers (p < 0.05). CONCLUSIONS: Therefore, mtDNA and associated mutations are present and detectable in serum and plasma. Plasma and serum might be alternative sources for the molecular diagnosis of mt3243 associated diabetes mellitus, as well as other mitochondrial mediated diseases.

Adult↗
PubMed2000

Plasma DNA as a prognostic marker in trauma patients.

BACKGROUND: Recently, much interest has developed in the potential use of plasma DNA as a diagnostic and monitoring tool. We hypothesized that plasma DNA is increased in patients with trauma and may be prognostic in such patients. METHODS: We studied 84 patients who had sustained an acute blunt traumatic injury. We measured plasma DNA by a real-time quantitative PCR assay for the beta-globin gene. Blood samples were collected at a median time of 60 min following injury. Blood samples were also obtained from 27 control subjects. RESULTS: The median plasma DNA concentrations in the control, minor/moderate trauma (Injury Severity Score <16; n = 47), and major trauma (Injury Severity Score > or =16; n = 37) groups were 3154 kilogenome-equivalents/L, 13 818 kilogenome-equivalents/L, and 181 303 kilogenome-equivalents/L, respectively. Plasma DNA concentrations in patients with adverse outcomes, including acute lung injury, acute respiratory distress syndrome, and death, had 11. 6- to 12-fold higher plasma DNA concentrations than those who did not develop these complications. At a cutoff of 232 719 kilogenome-equivalents/L, the sensitivities of plasma DNA analysis for the prediction of acute lung injury, acute respiratory distress syndrome, and death were 100% (95% confidence interval, 100-100%), 100% (95% confidence interval, 100-100%), and 78% (95% confidence interval, 40-97%), respectively. The respective specificities were 81% (95% confidence interval, 71-89%), 80% (95% confidence interval, 70-88%), and 82% (95% confidence interval, 71-90%). CONCLUSIONS: Plasma DNA is increased after trauma and may be a potentially valuable prognostic marker for these patients.

Biomarkers↗
PubMed2000

Quantitative analysis of the bidirectional fetomaternal transfer of nucleated cells and plasma DNA.

BACKGROUND: Recently, much interest has been generated on the fetomaternal transfer of nucleated cells and plasma DNA. However, there has been no systematic quantitative comparison of these two directions and two modalities of trafficking within the same study population. METHODS: The fetus-to-mother transfer of nucleated cells and plasma DNA in pregnant women carrying male babies was studied using a real-time quantitative PCR assay for the S:RY gene. For mother-to-fetus transfer, real-time quantitative PCR assays for the insertion/deletion polymorphisms involving the glutathione S:-transferase M1 and angiotensin-converting enzyme genes were used. RESULTS: Of the 50 informative mother-baby pairs, maternal DNA was detected in the cellular fraction of umbilical cord blood in 24% of cases (12 of 50), at a median fractional concentration of 2.6 x 10(-4) (interquartile range, 1.7 x 10(-4) to 3.6 x 10(-4)). In the plasma fraction of cord blood, maternal DNA was detected in 30% (15 of 50) of cases at a median fractional concentration of 3 x 10(-3) (interquartile range, 1 x 10(-3) to 1.6 x 10(-2)). For the other direction of trafficking, fetus-to-mother transfer of nucleated cells was detected in 26% of cases (13 of 50) at a median fractional concentration of 3.2 x 10(-4) (interquartile range, 0.6 x 10(-4) to 7.6 x 10(-4)). In the plasma fraction, fetal DNA was detected in 100% of maternal plasma (50 of 50) at a median fractional concentration of 3 x 10(-2) (interquartile range, 1.4 x 10(-2) to 5. 3 x 10(-2)). CONCLUSIONS: This study indicated that significantly more fetal DNA is present in the plasma of pregnant women compared with DNA from the cellular fraction of maternal blood. In addition, maternal DNA was demonstrated in both the cellular and plasma fractions of cord blood after delivery. This study has therefore determined the fundamental quantitative values for the bidirectional fetomaternal cellular and plasma DNA traffic.

Biological Transport↗
PubMed2000

Frequent p15 promoter methylation in tumor and peripheral blood from hepatocellular carcinoma patients.

We prospectively analyzed p15 methylation patterns in 25 surgically resected tumors and 130 plasma, serum, and buffy coat samples from hepatocellular carcinoma (HCC) patients, controls with chronic hepatitis/cirrhosis, and healthy subjects. Using methylation-specific PCR, we demonstrated for the first time p15 promoter methylation in 64% of tumors and 25% (4 of 16) of patients' plasma and serum samples. Concurrent p15 and p16 methylation was shown in 48% of tumors, and p15/p16 methylation was detected in the plasma/serum of 92% (11 of 12) of patients. Of note, 75% of 12 patients with concurrent tumor methylation developed clinical metastasis/recurrence (P = 0.027). In buffy coat samples, p15 methylation was detected in all eight patients with tumor p15 methylation, suggesting the presence of circulating tumor cells. None of the control samples were methylation positive. Our data underscore the important role(s) of p15 and p16 methylation in hepatocarcinogenesis and tumor progression. Among 92% (23 of 25) of patients with tumor p15/p16 methylation, circulating tumor DNA and HCC cells were detected in the peripheral blood of 87% (20 of 23) of patients. The combination of these epigenetic markers may prove valuable for noninvasive HCC diagnosis and disease monitoring.

Blotting, Southern↗
PubMed2000

Fetal DNA in maternal plasma: biology and diagnostic applications.

BACKGROUND: Molecular analysis of plasma DNA during human pregnancy has led to the discovery that maternal plasma contains both fetal and maternal DNA. This valuable source of fetal DNA opens up new possibilities for noninvasive prenatal diagnosis. APPROACH: Published data from the last 3 years demonstrating the feasibility and utility of analyzing fetal DNA in maternal plasma are reviewed. CONTENT: The detection of fetal DNA in maternal plasma is much simpler and more robust than detecting fetal nucleated cells in maternal blood, and does not require prior enrichment. This approach has been shown to have application in the prenatal diagnosis of fetal rhesus D status, sex-linked disorders, and other paternally inherited genetic disorders. Abnormal fetal DNA concentrations in maternal plasma and serum have been found in common pregnancy-associated disorders, including preterm labor and preeclampsia, as well as in pregnancies complicated by fetal trisomy 21. After delivery, fetal DNA is cleared rapidly from maternal plasma, with a half-life in the order of minutes. These clearance kinetics exhibit an important difference from fetal cell clearance, where long-term persistence has been demonstrated. SUMMARY: It has been only 3 years since fetal DNA was first detected in maternal plasma, and much remains to be learned about the biology of this phenomenon. In addition, additional diagnostic applications beyond those discussed here can be expected in the near future.

DNA↗