Experimental treatment of bilateral fetal chylothorax using in-utero pleurodesis.

OBJECTIVE: To assess the use and efficacy of in-utero pleurodesis for experimental treatment of bilateral fetal chylothorax. METHODS: This was a study of 78 fetuses with bilateral pleural effusion referred to three tertiary referral centers in Taiwan between 2005 and 2009. Fetuses were karyotyped following amniocentesis and the lymphocyte ratio in the pleural effusion was determined following thoracocentesis. Forty-nine (62.8%) fetuses had a normal karyotype and were recognized to have fetal chylothorax; of these, 45 underwent intrapleural injection of 0.1KE OK-432 per side per treatment. We evaluated clinical (hydrops vs. no hydrops) and genetic (mutations in the reported lymphedema-associated loci: VEGFR3, PTPN11, FOXC2, ITGA9) parameters, as well as treatment outcome. Long-term survival was defined as survival to 1 year of age. RESULTS: The overall long-term survival rate (LSR) was 35.6% (16/45); the LSR for non-hydropic fetuses was 66.7% (12/18) and for hydropic fetuses it was 14.8% (4/27). If we included only fetuses with onset of the condition in the second trimester, excluding those with onset in the third trimester, the LSR decreased to 29.4% (10/34). Notably, 29.6% (8/27) of hydropic fetuses had mutations in three of the four loci examined. CONCLUSIONS: OK-432 pleurodesis appeared to be an experimental alternative to the gold-standard technique of thoracoamniotic shunting in non-hydropic fetal chylothorax. In hydropic fetuses, pleurodesis appeared less effective.

Genetic evaluation and management of fetal chylothorax: review and insights from a case of Noonan syndrome.

Fetal chylothorax is one of a very few syndromes that can be treated in utero with thoracoamniotic shunting or pleurodesis by OK-432 as two major therapeutic modalities. We report on a fetus with Noonan syndrome and a missense mutation c.182A > C (p.Asp61Ala) of PTPN11 who responded poorly to antenatal pleurodesis by OK-432. Based on our previous publication and this case study, we propose that fetal chylothorax of a distinct genetic origin may respond poorly to OK-432 pleurodesis.

The spectrum of the factor 8 (F8) defects in Taiwanese patients with haemophilia A.

Haemophilia A (HA) is an X-linked recessive bleeding disorder caused by various types of pathological defects in the factor VIII gene (F8), which encodes coagulation factor VIII (FVIII). To date, several studies on the spectra of F8 defects have been performed in Western populations, but similar studies in Asian races are scarce. Here, we report the distribution of the mutations within the F8 gene in 31 Taiwanese unrelated HA patients (19 severe and 10 moderate/mild males and two severe females). Of these, 12 (38.7%) and one (3.2%) severe males were genotyped with the recurrent IVS22 and IVS1 inversion, respectively, similar to that in general populations (IVS22: 40-50%; IVS1: 2-5%). The F8 defects in the remaining 18 inversion-negative patients cover a wide spectrum, in which 17 different mutations were identified (10 missense and three nonsense mutations, and two small and two large deletions). Eleven of these mutations are novel: seven caused missense substitutions and four resulted in truncated proteins. To assess the putative pathogenetic impacts of the newly amino acid substitutions, computer analyses were performed based on molecular 3D modelling. The degree of conservation in cross-species FVIIIs and the position in known functional FVIII regions were studied. The novel missense mutations found in our series all occurred at evolutionary conserved residues that may carry a functional importance in our analyses. The results of this study add the short list of Taiwanese/Chinese F8 mutations, and will enhance our understanding of the molecular basis of FVIII function and the mechanism underlying HA.

Classification of benign and malignant breast tumors using neural networks and three-dimensional power Doppler ultrasound.

OBJECTIVES: To evaluate the use of three-dimensional (3D) power Doppler ultrasound in the differential diagnosis of solid breast tumors using a neural network model as a classifier. METHODS: Data from 102 benign and 93 malignant breast tumor images that had pathological confirmation were collected consecutively from January 2003 to February 2004. We used 3D power Doppler ultrasound to calculate three indices (vascularization index (VI), flow index (FI) and vascularization flow index (VFI)) for the tumor itself and for the tumor plus a 3-mm shell surrounding it. These data were applied to a multilayer perception (MLP) neural network model and we evaluated the model as a classifier to assess the capability of 3D power Doppler sonography to differentiate between benign and malignant solid breast tumors. RESULTS: The accuracy of the MLP model for classifying malignancy was 84.6%, the sensitivity was 90.3%, the specificity was 79.4%, the positive predictive value was 80.0% and the negative predictive value was 90.0%. When the neural network was used to combine the three 3D power Doppler indices, the area under the receiver-operating characteristics curve was 0.89. CONCLUSIONS: 3D power Doppler ultrasound may serve as a useful tool in distinguishing between benign and malignant breast tumors, and its capability may be increased by using a MLP neural network model as a classifier.

Computer-aided diagnosis using morphological features for classifying breast lesions on ultrasound.

OBJECTIVES: To develop and evaluate a computer-aided diagnosis (CAD) system with automatic contouring and morphological analysis to aid in the classification of breast tumors using ultrasound. METHODS: We evaluated 118 breast lesions (34 malignant and 84 benign tumors). Each tumor contour was automatically extracted from the digitized ultrasound image. Nineteen practical morphological features from the extracted contour were calculated and principal component analysis (PCA) was applied to find independent features. A support vector machine (SVM) classifier utilized the selected principal vectors to identify the breast tumor as benign or malignant. In this study, all the cases were sampled with k-fold cross-validation (k = 10) to evaluate the performance by receiver-operating characteristics (ROC) curve analysis. RESULTS: The areas under the ROC curves for the proposed CAD systems using all morphological features and the lower-dimensional principal vector were 0.91 and 0.90, respectively. The classification ability for breast tumors using morphological information was good. CONCLUSIONS: This system differentiates benign from malignant breast tumors well and therefore provides a clinically useful second opinion. Moreover, the morphological features are nearly setting-independent and thus available to various ultrasound machines.

The diagnostic utility of preoperative breast magnetic resonance imaging (MRI) and/or intraoperative sub-nipple biopsy in nipple-sparing mastectomy.

BACKGROUND: The necessity of routine sub-nipple biopsy was uncertain, and the role of preoperative magnetic resonance imaging (MRI) in detecting nipple invasion in patients who have been selected for nipple sparing mastectomy (NSM) has not been adequately evaluated. METHODS: We retrospectively collected and analyzed the medical and surgical records of 434 patients with primary operable breast cancer who met the criteria for NSM and underwent breast surgery during the period January 2011 to December 2015. Patients were stratified into three risk groups (low, intermediate, and high) according to tumor size and tumor-to-nipple distance. RESULTS: Among the 434 patients in this study, 29 (6.7%) had occult invasion of the nipple-areola complex (NAC). Sub-nipple biopsy had a sensitivity of 84.6%, a specificity of 100%, a false negative rate of 1.2%, a false positive rate of 0%, and an overall accuracy rate of 98.8% in confirming NAC invasion. The NAC invasion rate was 0% in the low-risk group, 5.1% in the intermediate-risk group, and 19.7% in the high-risk group (P < 0.01). The overall NPV of preoperative MRI for predicting NAC invasion was 94.8%. Cost analysis revealed that the cost of NSM with sub-nipple biopsy was significantly higher than that of NSM alone, with a mean difference in cost of USD 238.5 (P < 0.01). CONCLUSION: The high negative predictive value of MRI for NAC invasion is useful for selection of patients receiving NSM. Sub-nipple biopsy is a reliable procedure to detect occult NAC invasion, however, routine use is not cost-effect for low risk patients.

Personal experience of ultrasound-guided 14-gauge core biopsy of breast tumor.

AIM: This study was aimed to examine the efficacy of ultrasound-guided core needle biopsy of breast tumor and compared with the results of previous publications. METHODS: From January 2001 to September 2003, 546 lesions in 513 consecutive patients with the identification of a tumor on ultrasound examination categorized belong and above C3 according to BIRADS (Breast Image Reporting and Data Systems). A minimum of three samples was taken during the biopsy process. RESULTS: The patients' ages ranged from 17 to 89 years (mean, 43 years); tumors were from 5.7 to 41.6 mm in diameter (mean, 20.3 mm). There were 341 lesions with benign findings, 202 lesions with malignancy and 3 lesions with atypical ductal hyperplasia (ADH). Underestimates were found in five patients. Two patients were "ADH-DCIS (ductal carcinoma in situ) underestimate", and three patients were "DCIS-IDC (invasive ductal carcinoma) underestimate". The ADH underestimation rate was 40% (2/5) and DCIS underestimation rate was 43% (3/7). No false-positive results were observed. According to this study, the accuracy rate was 99%, sensitivity rate 97.6%, specificity rate 100%, and false-negative rate 2.4%. CONCLUSIONS: Both palpable and impalpable breast lesions should be examined under image guidance and automated core biopsy is the technique of first choice. Fourteen-gauge core biopsy can provide a definitive diagnosis in 99% of solid tumors in this series.

Objective and noninvasive detection of sub-clinical lung injury in breast cancer patients after radiotherapy.

AIMS: We employed technetium-99m hexamethylpropylene amine oxime (Tc-99m HMPAO) lung scan to detect sub-clinical lung injury after radiation therapy of 60 female patients diagnosed with right breast cancer. METHODS: The degree of pulmonary vascular endothelium damage was represented as lung/liver uptake ratios (L/L ratios) calculated on Tc-99m HMPAO lung scan. All patients underwent simple mastectomy and post-operative radiotherapy of approximately 50 Gy. We divided the patients into three groups according to the interval between radiotherapy and lung Tc-99m HMPAO lung scan: Group 1 included 20 patients who received the lung scan within 1-3 months after radiotherapy, group 2 included 20 patients were within 3-9 months after radiotherapy, and group 3 included 20 patients were more than 9 months after radiotherapy. In addition, 20 age-matched normal women were included as the control group. RESULTS: The L/L ratios were 0.32+/-0.04 for normal controls, 0.59+/-0.10 for group 1, 0.55+/-0.07 for group 2, and 0.34+/-0.04 for group 3, respectively. Based on our preliminary results, we found that sub-clinical lung injury and significantly increased L/L ratio in breast cancer patients received radiotherapy may occur within the first 6 months after radiotherapy. However, the L/L ratio is markedly decreased after 9 months. CONCLUSION: Our findings concluded that the degree of pulmonary vascular endothelium damage represented as the L/L ratio on Tc-99m HMPAO lung scan has the potential to be a sensitive, objective and noninvasive method to detect sub-clinical lung injury in breast cancer patients received radiotherapy.

Alteration in the expression of protein kinase C isoforms in human hepatocellular carcinoma.

This study was designed to investigate the alterations of individual protein kinase C (PKC) isoforms in human liver cancer. Surgical specimens of hepatocellular carcinoma and adjacent normal tissues were extracted into cytosolic and membranous fractions. The level of membrane-bound PKCalpha in the cancer tissue was significantly lower than that in the adjacent normal tissue and consistent with the change in PKC activity. In addition, there was a significant negative correlation between PKCalpha and tumor size. In both cytosolic and membrane fractions, levels of PKCdelta and PKCzeta was significantly higher in the cancer tissue than those in the adjacent normal liver tissue. The alterations in the PKC isoforms signify their roles in the hyperproliferation in liver cancer.

Prognostic significance of triple negative breast cancer at tumor size 1 cm and smaller.

AIMS: The purpose of this study was to clarify the prognostic significance of triple-negative breast cancer (TNBC) with a tumor size ≤ 1 cm. MATERIALS AND METHODS: Patients with primary operable breast cancer with a tumor size ≤ 1 cm were enrolled at Changhua Christian Hospital and National Cheng-Kung University Hospital. Tumors negative for ER, PR, and HER-2 were classified as TNBCs and compared with tumors with any receptor positivity (non-TNBC) for disease-free survival (DFS) and cancer-specific survival (CSS). RESULTS: From 1995 to 2006, a total of 377 patients with tumor size ≤ 1 cm were enrolled. Compared with non-TNBC patients, TNBC patients with a tumor size ≤ 1 cm as a whole or in a lymph node-positive subgroup were not associated with a poorer 5-year DFS and CSS. In lymph node-negative patients (pT1a-bN0M0), TNBC was associated with a poorer 5-year CSS but not DFS. Compared with the hormone receptor-positive, HER-2-negative subgroup, TNBC was associated with poorer DFS and CSS. In the multivariate Cox regression hazard analysis, lymph node invasion was the most important cause of disease recurrence and cancer-specific death. CONCLUSION: TNBC is very likely an independent risk factor in small (≤1 cm) node-negative invasive breast cancer. With tumors 1 cm and smaller, lymph node invasion was the single most important prognostic factor.

Capsaicin may induce breast cancer cell death through apoptosis-inducing factor involving mitochondrial dysfunction.

The majority of breast cancer patients are resistant to chemotherapy or radiotherapy due to the down-regulation or lack of caspase-3 expression. Capsaicin was found to inhibit cancer cell growth in caspase-3-deficient human breast cancer cells. This study aimed to investigate the growth-inhibitive effect of capsaicin and its mechanisms in human breast cancer cell lines, MCF-7 and BT-20. The results showed that cell viability decreased in a dose-dependent manner in both the caspase-3-deficient and non-deficient cells through inducing cell apoptosis and arresting the cell cycle in the S phase. Capsaicin significantly decreased mitochondria membrane potential, induced the cleavage of PARP-1, and decreased procaspase-7 expression in both cells. Apoptosis-inducing factor (AIF) was distinctly released from mitochondria and translocated into the cytoplasm and nucleus in MCF-7 cells (52.9%), but not in BT-20 cells (2%) after treatment with 200 µM of capsaicin for 24 hours. Capsaicin inhibited breast cancer cell growth through inducing cell apoptosis and cell cycle arrest in the S phase. This apoptotic effect could be induced through the mitochondrial pathway, and PARP-1 subsequently cleaved by activation of caspase-7. The application of capsaicin in clinical therapy could be useful for breast cancer patients.

The management of papillary lesion of the breast by core needle biopsy.

BACKGROUND: The purpose of this study was to determine the accuracy of core needle biopsy (CNB) diagnoses of papillary breast lesions, and to identify the risk factors and histologic features, compared with excisional biopsy (EB). METHOD: We retrospectively reviewed 1,682 sonographically guided core needle biopsies performed at one single medical center from January 2001 through December 2005, and identified 41 papillary lesions. Surgical correlation was available for 35 cases, 2 cases were loss follow-up and 4 cases diagnosed as papilloma by core needle biopsy were followed up with imaging for at least 24 months. RESULTS: The pathologic diagnoses for the 35 papillary lesions obtained at core biopsy were benign in 24 cases, atypical in 7, and malignant in 4. Of those diagnosed as benign lesions at CNB, 7/24 turned out to be malignant. In the hyperplasia group, 5/7 were malignant. The total upgrade rate was 39%. The positive predict value was 100% and negative predict value was 61%. CONCLUSION: All papillary lesions of the breast diagnosed by CNB should be excised because a substantial number of lesions were upgraded of diagnoses at excision.

Subtelomeric rearrangements and 22q11.2 deletion syndrome in anomalous growth-restricted fetuses with normal or balanced G-banded karyotype.

OBJECTIVE: To determine the frequencies of cryptic subtelomeric rearrangements and 22q11.2 deletion in anomalous growth-restricted fetuses with normal or balanced G-banded karyotypes. METHODS: This was a study of 27 consecutive fetuses at a median gestational age of 26 (range, 19-33) weeks, that had intrauterine growth restriction (IUGR) as well as at least one major structural anomaly, and a normal or balanced G-banded karyotype. The median maternal age was 29 (range, 17-39) years. Fluorescence z in-situ hybridization (FISH) diagnosis of the cultured amniocytes with the probe TUPLE 1, and then the Chromoprobe Multiprobe-T system were used, respectively, to screen for the frequency of 22q11.2 deletion syndrome and subtelomeric rearrangements involving the 41 unique chromosome ends (i.e. excluding the five short arms of acrocentric autosomes (no. 13, 14, 15, 21 and 22)). Those that had suspected deleted subtelomeres were reanalyzed with a specific subtelomeric probe, TelVysion. RESULTS: Of the 27 fetuses, three (11%) were affected with 22q11.2 deletion syndrome and two (7.4%) had subtelomeric deletions (one monosomy 21q22.3, one monosomy 1p36.3). Of the 11 fetuses with congenital heart defects, three (27.3%) had 22q11.2 deletion syndrome and one (9.1%) had monosomy 1p36.3. In the remaining 16 fetuses without congenital heart defects, none had 22q11.2 deletion syndrome. However, one (6.3%) had cryptic rearrangement involving subtelomeres. CONCLUSION: Prenatal subtelomeric FISH screening is technically feasible using cultured amniocytes. We propose that 22q11.2 deletion syndrome and cryptic subtelomere rearrangements may be important etiologies of fetuses with IUGR and at least one structural anomaly, along with a normal karyotype or one that is balanced by traditional G-banding. Fetuses with congenital heart defects and IUGR should undergo FISH to exclude 22q11.2 deletion syndrome. In fetuses with IUGR and at least one major structural anomaly but without congenital heart defects, screening of subtelomeric rearrangements may contribute to further elucidation of the underlying etiology.

Image retrieval with principal component analysis for breast cancer diagnosis on various ultrasonic systems.

OBJECTIVES: We present a computer-aided diagnostic (CAD) system with textural features and image retrieval strategies for classifying benign and malignant breast tumors on various ultrasonic systems. Effective applications of CAD have used different types of texture analysis. Nevertheless, most approaches performed in a specific ultrasonic machine do not indicate whether the technique functions satisfactorily for other ultrasonic systems. This study evaluated a series of pathologically proven breast tumors using various ultrasonic systems. METHODS: Altogether, 600 ultrasound images of solid breast nodules comprising 230 malignant and 370 benign tumors were investigated. All ultrasound images were acquired from four diverse ultrasonic systems. The suspicious tumor area in the ultrasound image was manually chosen as the region-of-interest (ROI) subimage. Textural features extracted from the ROI subimage are supported in classifying the breast tumor as benign or malignant. However, the textural feature always behaves as a high-dimensional vector. In practice, high-dimensional vectors are unsatisfactory at differentiating breast tumors. This study applied the principal component analysis (PCA) to project the original textural features into a lower dimensional principal vector that summarized the original textural information. The image retrieval techniques were employed to differentiate breast tumors, according to the similarities of the principal vectors. The query ROI subimages were identified as malignant or benign tumors according to characteristics of retrieved images from the ultrasound image database. RESULTS: Using the proposed CAD system, historical cases could be directly added into the database without a retraining program. The area under the receiver-operating characteristics curve for the system was 0.970+/-0.006. CONCLUSION: The CAD system identified solid breast nodules with comparatively high accuracy in the different ultrasound systems investigated.

Evaluation of the CO2 laser in performance of breast surgery.

BACKGROUND: In several studies the CO2 laser has proved to have properties which make it particularly useful to the general surgeon in management of breast tumor. METHODS: Four hundred and seventy-two patients, including 2 males, which age range from 15 to 84 years. The operative procedures included excisional biopsy, segmentectomy and modified radical mastectomy. In this study, one group was operated on by the scalpel; for the other group CO2 Laser (Sharplan Model 733) was used. This is a comparison of the healing process, the operative course and follow-up of patients who underwent breast surgery. Eighteen of this series were fresh, locally advanced mammary carcinoma cases and were managed with induction chemotherapy using cyclophosphamide, adriamycin, 5-fluorouracil, followed by definitive extirpative laser surgery. RESULTS: The patient have been followed up from 12 to 60 months. Good cosmetic results without local recurrence of tumor were observed following the laser treatment. An effective massive tumor necrosis of 80% occurred after induction chemotherapy. Swelling and distortion of cancer cells with vesicles filling in the cytoplasm and nucleus were demonstrated. CONCLUSIONS: Laser technology for breast surgery was concluded to assist the operating surgeon with more precision, smaller wounds, more hemostasis, less tissue damage, and reduced morbidity except prolonged wound healing.

Chilling-Induced Lipid Degradation in Cucumber (Cucumis sativa L. cv Hybrid C) Fruit.

Chilling at 4 degrees C in the dark induced lipid degradation in cucumber (Cucumis sativa L.) fruit upon rewarming at 14 degrees C. Rates of ethane evolution by fruits rewarmed after 3 days of chilling were up to four-fold higher than those evolved by unchilled (14 degrees C) fruits (0.02-0.05 picomoles gram fresh weight(-1) hour(-1)). This potentiation of lipid peroxidation occurred prior to irreversible injury (requiring 3 to 7 days of chilling) as indicated by increases in ethylene evolution and visual observations. Decreases in unsaturation of peel tissue glycolipids were observed in fruits rewarmed after 3 days of chilling, indicating the plastids to be the site of the early phases of chilling-induced peroxidation. Losses in unsaturation of tissue phospholipids were first observed only after chilling for 7 days. Phospholipase D activity appeared to be potentiated in fruits rewarmed after 7 days of chilling as indicated by a decrease in phosphatidylcholine (and secondarily phosphatidylethanolamine) with a corresponding increase in phosphatidic acid. These results indicate that lipid peroxidation may have a role in conferring chilling injury.

Single-particle mass spectrometry of polystyrene microspheres and diamond nanocrystals.

High-resolution mass spectra of single submicrometer-sized particles are obtained using an electrospray ionization source in combination with an audio frequency quadrupole ion-trap mass spectrometer. Distinct from conventional methods, light scattering from a continuous Ar-ion laser is detected for particles ejected out of the ion trap. Typically, 10 particles are being trapped and interrogated in each measurement. With the audio frequency ion trap operated in a mass-selective instability mode, analysis of the particles reveals that they all differ in mass-to-charge ratio (m/z), and the individual peak in the observed mass spectrum is essentially derived from one single particle. A histogram of the spectra acquired in 10(2) repetitions of the experiment is equivalent to the single spectrum that would be observed when an ion ensemble of 10(3) particles is analyzed simultaneously using the single-particle mass spectrometer (SPMS). To calibrate such single-particle mass spectra, secular frequencies of the oscillatory motions of the individual particle within the trap are measured, and the trap parameter qz at the point of ejection is determined. A mass resolution exceeding 10(4) can readily be achieved in the absence of ion ensemble effect. We demonstrate in this work that the SPMS not only allows investigations of monodisperse polystyrene microspheres, but also is capable of detecting diamond nanoparticles with a nominal diameter of 100 nm, as well.