A prenatal diagnosis of partial monosomy 18p(18p11.2-->pter) and trisomy 21q(21q22.3-->qter) in a fetus with alobar holoprosencephaly (HPE) and premaxillary agenesis (PMA) but without the classical Down syndrome phenotype is reported. A 27-year-old primigravida woman was referred for genetic counselling at 21 weeks' gestation due to sonographic findings of craniofacial abnormalities. Level II ultrasonograms manifested alobar HPE and median orofacial cleft. Cytogenetic analysis and fluorescence in situ hybridization (FISH) on cells obtained from amniocentesis revealed partial monosomy 18p and a cryptic duplication of 21q,46,XY,der(18)t(18;21)(p11.2;q22.3), resulting from a maternal t(18;21) reciprocal translocation. The breakpoints were ascertained by molecular genetic analysis. The pregnancy was terminated. Autopsy showed alobar HPE with PMA, pituitary dysplasia, clinodactyly and classical 18p deletion phenotype but without the presence of major typical phenotypic features of Down syndrome. The phenotype of this antenatally diagnosed case is compared with those observed in six previously reported cases with monosomy 18p due to 18;21 translocation. The present study is the first report of concomitant deletion of HPE critical region of chromosome 18p11.3 and cryptic duplication of a small segment of distal chromosome 21q22.3 outside Down syndrome critical region. The present study shows that cytogenetic analyses are important in detecting chromosomal aberrations in pregnancies with prenatally detected craniofacial abnormalities, and adjunctive molecular investigations are useful in elucidating the genetic pathogenesis of dysmorphism.

Adult↗

PubMed2001

Rapid monitoring assay of congenital adrenal hyperplasia with microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots.

17-hydroxyprogesterone (17OHP) is the most important plasma parameter for diagnosing and monitoring congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency. A rapid, simple, and specific method based on microbore high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (micro-HPLC/ESI-MS/MS) was developed to determine the presence of 17OHP on dried filter-paper blood samples from patients with CAH caused by 21-hydroxylase deficiency. 17OHP from dried blood spots formed by the action of Girard reagent P (GirP) turned out to be a water-soluble hydrazone complex. Derivatization with GirP led to higher ESI sensitivity for 17OHP. The LC/MS/MS detection of GirP-derivatized 17OHP (GirP-17OHP) was rapid (<3 min). The method is repeatable and reproducible, with CVs <7% and 12%, respectively. This new method was used for direct determination of 17OHP in dried blood specimens obtained from abnormal children and infants of various ages with a detection limit of 10 ng/mL ( approximately 12 microL blood). The method described allows for rapid and reliable measurements of 17OHP in dried blood specimens from patients affected by CAH.

17-alpha-Hydroxyprogesterone↗

PubMed2001

High-resolution cranial ultrasound in the shaken-baby syndrome.

With limited near-field resolution and accessible acoustic windows, sonography has not been advocated for assessing central nervous system injuries in the shaken-baby syndrome. Our purpose was to correlate high-resolution ultrasonographic characteristics of central nervous system injuries in whiplash injuries and the shaken-baby-syndrome with MRI and CT. Ultrasonographic images of 13 infants, aged 2-12 months, with whiplash or shaking cranial trauma were reviewed and compared with MRI in 10 and CT in 10. Five patients had serial ultrasonography and MRI or CT follow-up from 1 to 4 months after the initial injury. With ultrasonography we identified 20 subdural haematomas. MRI and CT in 15 of these showed that four were hyperechoic in the acute stage, three were mildly echogenic in the subacute stage, and that one subacute and seven chronic lesions were echo-free. Five patients had acute focal or diffuse echogenic cortical oedema which evolved into subacute subcortical hyperechoic haemorrhage in four, and well-defined chronic sonolucent cystic or noncystic encephalomalacia was seen at follow-up in two. Using ultrasonography we were unable to detect two posterior cranial fossa subdural haematomas or subarachnoid haemorrhage in the basal cisterns in three cases, but did show blood in the interhemispheric cistern and convexity sulci in two. U1-trasonography has limitations in demonstrating abnormalities remote from the high cerebral convexities but may be a useful adjunct to CT and MRI in monitoring the progression of central nervous system injuries in infants receiving intensive care.

Battered Child Syndrome↗

PubMed2001

Bone mineral density in young female Chinese dancers.

To evaluate of the effect of dancing on bone mineral density (BMD) we compared 29 Chinese girls who had been receiving regular ballet training for at least 6 years with a control group of 20 nonathletic sex- and age-matched Chinese girls. BMD was measured at the second to fourth lumbar spine and at the right femoral neck using a commercial dual-energy X-ray absorptiometer (DEXA). There were no significant differences in either measured BMD or spinal BMD after adjusting for body weight (body mass index). Adjusted BMD of the femoral neck, however, showed significant differences (P <0.05). Results revealed that the ballet dancers were significantly thinner than the control group and the negative effect of a lower bodyweight may neutralize the positive effect of ballet dancing on BMD, especially in the femoral neck.

Adolescent↗

PubMed2001

Correlation between P-glycoprotein (P-gp) expression in parathyroid and Tc-99m MIBI parathyroid image findings.

The major factor to influence localization of parathyroid adenomas is tumor size. P-glycoprotein (P-gp) expression in parathyroid adenomas has been considered to be an another possible factor to influence localization of parathyroid adenomas because false-negative studies have been reported with large tumors and true-positives reported with very small tumors in previous studies. The aim of this study was to characterize Tc-99m MIBI uptake and retention by parathyroid adenomas and to correlate this with cell surface expression of P-gp. Sixteen patients with parathyroid adenoma (larger than 1.5 gm) underwent dual-phase (10min and 2hr) Tc-99m MIBI parathyroid image immediately before parathyroid exploration. Tissues were obtained from normal and abnormal parathyroid glands and from the thyroid gland. Immunohistochemistry (IHC) was obtained with monoclonal antibodies to identify P-gp expression in all tissues. All of the 16 parathyroid adenomas and 32 normal control specimens (16 normal parathyroid and 16 normal thyroid specimens) were submitted for P-gp detection by IHC. The dual-phase Tc-99m MIBI parathyroid image accurately localized 14 parathyroid adenomas, but not the remaining 2 adenomas. The 14 parathyroid adenomas with significant Tc-99m MIBI uptake in delayed 2hr images revealed negative P-gp expression, but the 2 adenomas without significant Tc-99m MIBI uptake, as well as normal parathyroid and normal thyroid specimens, revealed positive P-gp expression when evaluated by IHC. Not only the size of parathyroid adenomas, but also significant P-gp expression limited the sensitivity of dual-phase Tc-99m MIBI parathyroid image to localize parathyroid adenomas before operation.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

Data mining with decision trees for diagnosis of breast tumor in medical ultrasonic images.

To increase the ability of ultrasonographic (US) technology for the differential diagnosis of solid breast tumors, we describe a novel computer-aided diagnosis (CADx) system using data mining with decision tree for classification of breast tumor to increase the levels of diagnostic confidence and to provide the immediate second opinion for physicians. Cooperating with the texture information extracted from the region of interest (ROI) image, a decision tree model generated from the training data in a top-down, general-to-specific direction with 24 co-variance texture features is used to classify the tumors as benign or malignant. In the experiments, accuracy rates for a experienced physician and the proposed CADx are 86.67% (78/90) and 95.50% (86/90), respectively.

Breast Neoplasms↗

PubMed2001

An E. coli expression system for the extracellular secretion of barley alpha-amylase.

Libraries of modified genes are often screened during the process of genetically engineering enzymes with specifically tailored activities. It is important, therefore, to create expression systems which allow for the rapid screening of many clones. We developed an Escherichia coli expression system which will secrete enzymes into the growth medium. We describe the first reported expression of barley alpha-amylase in E. coli. The enzyme is secreted onto solid media containing starch to produce easily visualized halos. In addition, the enzyme is secreted into liquid media in an intact, active form.

Culture Media↗

PubMed2001

A novel topoisomerase II poison GL331 preferentially induces DNA cleavage at (C/G)T sites and can cause telomere DNA damage.

PURPOSE: Topoisomerase II (Topo II) preferentially cuts DNA at alternating purine-pyrimidine repeats. Different Topo II poisons may affect Topo II to produce distinct drug-specific DNA cleavage patterns. GL331 is a new podophyllotoxin derivative exhibiting potent Topo II-poisoning activity. Therefore, the sequence selectivity of GL331-induced DNA cleavage was determined. METHODS: Human gastric adenocarcinoma SC-M1 cells were treated with GL331, and the resultant DNA fragments were isolated by SDS-K+ precipitation. These DNA fragments were further cloned and sequenced to exhibit GL331-induced DNA cleavage sites. In addition, the telomere damage was detected by Southern blot analyses using a (TTAGGG)4 probe. GL331's effect on telomerase was examined using the TRAP assay. RESULTS: The selective sequences of GL331-induced DNA cleavage were analyzed. The first nucleotide 3'-terminal to the cleavage sites was preferentially C or G and followed by the second nucleotide T. More than 50% of GL331-induced DNA cleavage fragments exhibited AT-rich sequences in the first 20 nucleotides. In addition, the telomeric damage was observed both from GL331-treated SC-M1 cells and in vitro incubation of genomic DNA with GL331 and purified human Topo II. Although GL331 treatment reduced cellular telomerase activity, in vitro reaction data suggested that GL331 was not a telomerase inhibitor. CONCLUSION: GL331 preferentially induced Topo II-mediated DNA cleavage at (C/G)T sites. Because the telomeric repeat sequence contains GL331's GT preference site, the telomere was identified as one of the targets of GL331-induced DNA damage.

Antineoplastic Agents, Phytogenic↗

PubMed2001

Technetium-99m methoxyisobutylisonitrile chest imaging for small-cell lung cancer. Relationship to chemotherapy response (six courses of combination of cisplatin and etoposide) and p-glycoprotein or multidrug resistance related protein expression.

AIMS: This is a retrospective and adaptive randomization study. The purpose of this study was to evaluate the relationship between technetium-99m methoxyisobutylisonitrile (Tc-99m MIBI) chest-imaging results, chemotherapy response and P-glycoprotein (Pgp) or multidrug resistance related protein (MRP) expression in small-cell lung cancer (SCLC). PATIENTS AND METHODS: Before chemotherapy, 30 patients (11 females, 19 males, ages: 52-69 years) with SCLC, including 14 extensive diseases without localized problems and 16 limited diseases in excess of solitary pulmonary nodule, underwent early chest imaging, including visual interpretation and quantitative analyses of tumor uptake ratio (TUR), 10 minutes after intravenous injection of Tc-99m MIBI. Immunohistochemical analyses were performed, using multiple nonconsecutive sections of the biopsy specimens, to detect Pgp and MRP expressions. Chemotherapy response was evaluated in the third month after completion of treatment by clinical and radiological methods. RESULTS: All 15 (100%) of the SCLC patients with complete or partial response had positive Tc-99m MIBI chest SPECT results, but negative ones for both Pgp and MRP expression. Twelve of the 15 (80%) SCLC patients with no response or progressive disease had negative Tc-99m MIBI chest SPECT results and were positive for either Pgp or MRP expression (P < 0.05). Negative Tc-99m MIBI chest SPECT results predicted complete or partial response. The TUR of patients with complete or partial response (1.91 +/- 0.29 with a 95% confidence interval (95% CI): 1.75-2.07) was significantly higher than that of patients with no response or progressive disease (1.19 +/- 0.28 with a 95% CI: 1.04-1.35). CONCLUSION: Tc-99m MIBI chest images are a potential tool for understanding Pgp and MRP expressions in SCLC and for predicting patient chemotherapy response.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

Evaluation of five commercial serological tests for the detection of Helicobacter pylori infection in Chinese.

BACKGROUND: Commercial serological tests for the detection of Helicobacter pylori infection must be locally validated. We evaluated the accuracy of five commercial tests in the Chinese population. METHODS: Serum samples were collected from patients referred for upper endoscopy. Antral biopsies were taken for histological examination and culture of H. pylori. The gold standard for diagnosing H. pylori infection was positive histological staining and/or positive H. pylori culture. The serum samples were tested for H. pylori antibodies using the following tests: (i) Cobas Core Anti-H. pylori EIA; (ii) GAP IgG; (iii) GAP IgM; (iv) H. pylori microwell EIA (Quidel); and (v) Premier H. pylori. The sensitivity, specificity and accuracy of each test was calculated according to the manufacturers' instructions or according to a new cut-off value. RESULTS: A total of 158 patients were recruited amongst whom 114 (72%) were H. pylori-positive. Indeterminate results varied from 7% to 19%. The accuracy of the tests varied from 57% to 85%. By using new cut-off values, the accuracy was much improved, ranging from 73.4% to 86.7%. CONCLUSIONS: By defining new cut-off values for the Chinese population, we were able to improve the performance of some of the serology tests. This illustrates the importance of local validation.

Adolescent↗

PubMed2001

Mutation analysis of Crouzon syndrome and identification of one novel mutation in Taiwanese patients.

BACKGROUND: Crouzon syndrome is an autosomal dominant disorder causing premature fusion of the cranial suture. Mutations have been reported in exon IIIa or IIIc of the fibroblast growth factor receptor 2 (FGFR2) gene. METHODS: In the present study, nine unrelated Crouzon syndrome patients were screened for mutations in the two exons of FGFR2 by polymerase chain reaction and direct sequencing. RESULTS: Mutations were detected in 67% (6/9) of all cases. More than half the studied Crouzon patients carried a mutation resulting in either the loss or gain of a cysteine residue. A novel mutation, Tyr281Cys substitution, was discovered at exon IIIa. CONCLUSIONS: The mechanisms by which the same genotypes cause different phenotypes for each type of craniosynostosis syndrome in still uncertain. However, the molecular identification of the FGFR gene has made a great impact on the clinical classification of craniosynostosis syndromes; a new classification based on genotypes seems to be unavoidable.

Asian People↗