Novel human-derived EML4-ALK fusion cell lines identify ribonucleotide reductase RRM2 as a target of activated ALK in NSCLC.

INTRODUCTION: Echinoderm microtubule-associated protein-like 4 (EML4)-Anaplastic Lymphoma Kinase (ALK) rearrangements occur in 3% to 7% of lung adenocarcinomas and are targets for treatment with tyrosine kinase inhibitors (TKIs). Here we have developed three novel EML4-ALK-positive patient-derived Non-Small-Cell-Lung-Cancer (NSCLC) cancer cell lines, CUTO8 (variant 1), CUTO9 (variant 1) and CUTO29 (variant 3) and included a fourth ALK-positive cell line YU1077 (variant 3) to study ALK-positive signaling and responses. Variants 1 and 3 are the most common EML4-ALK variants expressed in ALK-positive NSCLC, and currently cell lines representing these EML4-ALK variants are limited. MATERIALS AND METHODS: Resazurin assay was performed to evaluate cell viability. Protein levels were determined using western blotting. RNA sequencing was performed in all four cell lines to identify differentially expressed genes. Whole-genome sequencing was performed to determine the presence of EML4-ALK fusion and ALK tyrosine kinase inhibitor resistance mutations. RESULTS: In this study, we have confirmed expression of the corresponding ALK fusion protein and assessed their sensitivity to a range of ALK tyrosine kinase inhibitors. These patient derived cell lines exhibit differential sensitivity to lorlatinib, brigatinib and alectinib, with EML4-ALK variant 3 containing cell lines exhibiting increased sensitivity to lorlatinib and brigatinib as compared to alectinib. These cell lines were further characterized by whole genome sequencing and RNA-seq analysis that identified the ribonucleotide reductase regulatory subunit 2 (RRM2) as a downstream and potential therapeutic target in ALK-positive NSCLC. CONCLUSION: We provide a characterization of four novel EML4-ALK-positive NSCLC cell lines, highlighting genomic heterogeneity and differential responses to ALK TKI treatment. The RNA-Seq characterization of ALK-positive NSCLC CUTO8, CUTO9, CUTO29 and YU1077 cell lines reported here, has been compiled in an interactive ShinyApp resource for public data exploration (https://ccgg.ugent.be/shiny/nsclc_rrm2_2022/).

[A retrospective analysis of clinical characteristics and outcomes of heart failure patients with different left ventricular ejection fractions].

Objective: To compare the clinical characteristics, and outcomes of patients with heart failure with different left ventricular ejection fractions (LVEF). Methods: A total of 1 182 hospitalized patients with heart failure (HF) were enrolled and retrospectively studied in the present study. The patients were stratified by LVEF as reduced (HFrEF, LVEF<40%, n=313), mid-range (HFmrEF, 40% ≤LVEF <50%, n=287) and preserved (HFpEF, LVEF≥50%, n=582) ejection fraction groups. Among the 1 182 cases, 941 of them (81.3%, 84.9%, and 84.0% inHFrEF, HFmrEF and HFpEF groups, respectively) were followed up for an median duration of 27.3 months. Results: (1) Among the study patients, 26.5% were in HFrEF, 24.3% in HFmrEF, and 49.2% in HFpEF groups. (2) Ischemic heart disease with HFmrEF was more frequent than that in patients with HFrEF. The average age, percentage of female subjects, systolic blood pressure, uric acid, N terminal B-type natriuretic peptide precursor (NT-proBNP), hemoglobin, and the incidence of hypertensive heart disease, anemia, atrial fibrillation in patients with HFmrEF were higher than those in patients with HFrEF, but lower than those in patients with HFpEF (all P<0.01). (3) The all-cause cumulative mortality was 10.8% at 1 year, 20.6% at 2 years and 35.9% at 5 years. No difference was observed in the all-cause cumulative mortality at 1 year, 2 years, 5 years among the three groups (all P>0.05). Conclusions: The HFmrEF patients, as a new and distinct group, were with many intermediate characteristics compared with HFrEF and HFpEF subjects. However, the all-cause mortality was not significantly different among HF patients with different LVEF.

[Validation of the Omron HEM-650 wrist blood pressure device using the British Hypertension Society protocol in emergency patients in Hong Kong]. / Validación del esfingomanómetro de muñeca Omron HEM-650 utilizando el protocolo de la Sociedad Británica de Hipertensión en pacientes atendidos en un servicio de urgencias de Hong Kong.

OBJECTIVES: Automated wrist cuff blood pressure (BP) devices are more compact and easier to use, particularly when access to the upper arm is restricted, for example in emergencies. MATERIAL AND METHODS: We tested the Omron HEM-650 wrist device using the validation criteria of the British Hypertension Society (BHS) protocol in a major emergency department (ED) in Hong Kong. 85 patients had three measurements each by both the Omron HEM-650 wrist device and the mercury sphygmomanometer. The conventional automated BP with arm cuff was also measured using an oscillometric (Colin BP-88S NXT) device for comparison. RESULTS: The Omron HEM-650 achieved a grade B for both systolic and diastolic BP and demonstrated acceptable accuracy and reliability in Chinese patients in the emergency setting. CONCLUSION: The Omron HEM 650 wrist device can be recommended for use in adult emergency patients. Further research is warranted for its use in pregnant women and critically ill patients.

OBJETIVO: Los dispositivos automáticos de muñeca para medir la presión arterial (PA) son más compactos y fáciles de usar, sobre todo cuando el acceso a la parte superior del brazo está restringido, por ejemplo, en situaciones de emergencia. METODO: Hemos probado el dispositivo de muñeca Omron HEM-650 utilizando los criterios de validación del protocolo de la Sociedad Británica de Hipertensión en un gran servicio de urgencias (ED) de Hong Kong. Se realizaron 3 mediciones en 85 pacientes con el dispositivo Omron HEM-650 y el esfingomanómetro de mercurio. También se utilizó la medición automatizada convencional de PA con banda para el brazo mediante un dispositivo oscilométrico (Colin BP-88S NXT) para la comparación. RESULTADOS: El Omron HEM-650 logra una calificación de grado B para las presiones arteriales sistólica y diastólica y demostró una precisión y fiabilidad aceptables en pacientes chinos en el servicio de urgencias. CONCLUSIONES: El dispositivo de muñeca Omron HEM 650 se recomienda para su uso en pacientes adultos de urgencias. Se necesita más investigación para su uso en mujeres embarazadas y los pacientes en estado crítico.

The synergistic effects of fibroblast growth factor-2 and mineral trioxide aggregate on an osteogenic accelerator in vitro.

AIM: To examine the effects of mineral trioxide aggregate (MTA)/fibroblast growth factor-2 (FGF-2) on material properties and in vitro human dental pulp cell (hDPCs) behaviour. METHODOLOGY: The setting time and diametral tensile strength (DTS) of MTA and MTA/FGF-2 were measured. The structure of specimens before and after soaking in DMEM was examined under a scanning electron microscope. Alamar Blue was used for evaluating hDPCs proliferation. An enzyme-linked immunosorbent assay was employed to determine ALP and osteocalcin (OC) expression in hDPCs cultured on cements. The effect of small interfering RNA (siRNA) transfection targeting fibroblast growth factor receptor (FGFR) was also evaluated. One-way analysis of variance was used to evaluate the significance of the differences between the mean values. RESULTS: Setting time and DTS data were not found to be significant (P > 0.05) between MTA with and without FGF-2. Cell proliferation and differentiation increased significantly (P < 0.05) with FGF-2 mixed MTA. After siRNA transfection with FGFR, the proliferation and differentiation behaviour of the hDPCs appreciably decreased when cultured on an MTA/FGF-2 composite. In contrast, no significant amounts (P > 0.05) of ALP and OC were secreted by hDPCs seeded on MTA. CONCLUSIONS: Mineral trioxide aggregate with FGF-2 content enhanced the higher expression of hDPCs proliferation and osteogenic differentiation as compared to pure MTA cement.

High power efficiency phosphorescent poly(dendrimer) OLEDs.

We show that it is possible to produce an efficient solution-processable phosphorescent poly(dendrimer) OLED with a 32 lm/W power efficiency at 100 cd/m2 without using a charge transporting host or any improvements in light extraction. This is achieved by using the dendrimer architecture to control inter-chromophore interactions. The effects of using 4,4',4″-tris(N-carbazolyl)triphenylamine (TCTA) as a charge transporting host and using a double dendron structure to further reduce inter-chromophore interactions are also reported.

Testosterone therapy delays cardiomyocyte aging via an androgen receptor-independent pathway

The testicular feminized (Tfm) mouse carries a nonfunctional androgen receptor (AR) and reduced circulating testosterone levels. We used Tfm and castrated mice to determine whether testosterone modulates markers of aging in cardiomyocytes via its classic AR-dependent pathway or conversion to estradiol. Male littermates and Tfm mice were divided into 6 experimental groups. Castrated littermates (group 1) and sham-operated Tfm mice (group 2, N = 8 each) received testosterone. Sham-operated Tfm mice received testosterone in combination with the aromatase inhibitor anastrazole (group 3, N = 7). Castrated littermates (group 4) and sham-operated untreated Tfm mice (group 5) were used as controls (N = 8 and 7, respectively). An additional control group (group 6) consisted of age-matched non-castrated littermates (N = 8). Cardiomyocytes were isolated from the left ventricle, telomere length was measured by quantitative PCR and expression of p16INK4α, retinoblastoma (Rb) and p53 proteins was detected by Western blot 3 months after treatment. Compared with group 6, telomere length was short (P < 0.01) and expression of p16INK4α, Rb and p53 proteins was significantly (P < 0.05) up-regulated in groups 4 and 5. These changes were improved to nearly normal levels in groups 1 and 2 (telomere length = 0.78 ± 0.05 and 0.80 ± 0.08; p16INK4α = 0.13 ± 0.03 and 0.15 ± 0.04; Rb = 0.45 ± 0.05 and 0.39 ± 0.06; p53 = 0.16 ± 0.04 and 0.13 ± 0.03), but did not differ between these two groups. These improvements were partly inhibited in group 3 compared with group 2 (telomere length = 0.65 ± 0.08 vs 0.80 ± 0.08, P = 0.021; p16INK4α = 0.28 ± 0.05 vs 0.15 ± 0.04, P = 0.047; Rb = 0.60 ± 0.06 vs 0.39 ± 0.06, P < 0.01; p53 = 0.34 ± 0.06 vs 0.13 ± 0.03, P = 0.004). In conclusion, testosterone deficiency contributes to cardiomyocyte aging. Physiological testosterone can delay cardiomyocyte aging via an AR-independent pathway and in part by conversion to estradiol.

Faecal occult blood screening: knowledge, attitudes, and practice in four Hong Kong primary care clinics.

OBJECTIVES. To assess primary care patients for their awareness, knowledge, and attitude towards colorectal cancer and screening, to report on the uptake of faecal occult blood test screening and the results of screening, and explore predictors of screening uptake. DESIGN. Cross-sectional study. SETTING. Four primary care clinics in Hong Kong. PATIENTS. A total of 1664 patients aged 50 to 74 years attending the clinics in the period July 2006 to July 2007. MAIN OUTCOME MEASURES. Percentage of subjects who were aware that colorectal cancer is common and curable at an early stage, and who knew that faecal occult blood test or colonoscopy is useful for screening; relevant knowledge score; uptake rate of faecal occult blood testing; rate of testing positive; and factors predicting uptake. RESULTS. A total of 1645 questionnaires were collected. In all, 89% (95% confidence interval, 88-91%) were aware that colorectal cancer is common, 95% (94-96%) believed faecal occult blood test and colonoscopy are useful for screening, and 58% (56-61%) achieved a knowledge score of 50% or above. The uptake rate of the faecal occult blood test was 35%. Uptake was higher among those with a positive family history (odds ratio=1.57; 95% confidence interval, 1.08-2.27; P=0.02), those who were more aware that colorectal cancer is common (1.86; 1.29-2.69; P=0.001), and that colorectal cancer is potentially curable at an early stage (1.76; 1.32-2.36; P=0.0001). Rate of testing positive was 2.1% (95% confidence interval, 0.9-3.3%); no colorectal cancer was detected and the neoplasia detection rate (for cancers and adenomas) was 5.1 per 1000 subjects screened. CONCLUSIONS. Patients were aware that colorectal cancer is common in our community, and faecal occult blood test or colonoscopy is useful for screening. The uptake of screening was low, though relatively higher for those with a positive family history and greater awareness of the high frequency and potential for cure of colorectal cancer. Faecal occult blood test positivity rate was 2.1%, and neoplasia detection rate 5.1 per 1000 screened.

Testosterone therapy delays cardiomyocyte aging via an androgen receptor-independent pathway.

The testicular feminized (Tfm) mouse carries a nonfunctional androgen receptor (AR) and reduced circulating testosterone levels. We used Tfm and castrated mice to determine whether testosterone modulates markers of aging in cardiomyocytes via its classic AR-dependent pathway or conversion to estradiol. Male littermates and Tfm mice were divided into 6 experimental groups. Castrated littermates (group 1) and sham-operated Tfm mice (group 2, N = 8 each) received testosterone. Sham-operated Tfm mice received testosterone in combination with the aromatase inhibitor anastrazole (group 3, N = 7). Castrated littermates (group 4) and sham-operated untreated Tfm mice (group 5) were used as controls (N = 8 and 7, respectively). An additional control group (group 6) consisted of age-matched non-castrated littermates (N = 8). Cardiomyocytes were isolated from the left ventricle, telomere length was measured by quantitative PCR and expression of p16INK4α, retinoblastoma (Rb) and p53 proteins was detected by Western blot 3 months after treatment. Compared with group 6, telomere length was short (P < 0.01) and expression of p16INK4α, Rb and p53 proteins was significantly (P < 0.05) up-regulated in groups 4 and 5. These changes were improved to nearly normal levels in groups 1 and 2 (telomere length = 0.78 ± 0.05 and 0.80 ± 0.08; p16INK4α = 0.13 ± 0.03 and 0.15 ± 0.04; Rb = 0.45 ± 0.05 and 0.39 ± 0.06; p53 = 0.16 ± 0.04 and 0.13 ± 0.03), but did not differ between these two groups. These improvements were partly inhibited in group 3 compared with group 2 (telomere length = 0.65 ± 0.08 vs 0.80 ± 0.08, P = 0.021; p16INK4α = 0.28 ± 0.05 vs 0.15 ± 0.04, P = 0.047; Rb = 0.60 ± 0.06 vs 0.39 ± 0.06, P < 0.01; p53 = 0.34 ± 0.06 vs 0.13 ± 0.03, P = 0.004). In conclusion, testosterone deficiency contributes to cardiomyocyte aging. Physiological testosterone can delay cardiomyocyte aging via an AR-independent pathway and in part by conversion to estradiol.

Clinical characteristics and treatment outcomes of patients undergoing nail avulsion surgery for dystrophic nails.

OBJECTIVE: To evaluate the clinical characteristics and treatment outcomes of patients undergoing surgical nail avulsion. DESIGN: Retrospective study. SETTING: Two dermatology centres in Hong Kong. PATIENTS: A total of 32 patients with nail diseases who underwent 33 nail avulsion procedures were reviewed from case records. MAIN OUTCOME MEASURES: Age, gender, co-morbidities, disease duration, clinical features, histopathology and fungal culture of nail plate, nail bed specimen for fungal culture in appropriate cases, and postoperative outcome. RESULTS: The mean age of the patients at the time of nail avulsion was 54 (range, 27-86) years. The most frequent preoperative findings were thickened nails (23 specimens, 70%) and discolouration (20 specimens, 61%). Onychomycosis was the most common pre-consultative diagnosis (20 specimens, 61%). Prior to nail avulsion, topical or systemic treatment had been tried in more than half of the cases. Histopathology of the avulsed nails confirmed onychomycosis in 24 (73%) of specimens. The clinical cure rate was 88% and the mycologic cure rate was 100%. The procedure was well tolerated without significant complications. Relapse was only noted in three (9%) of the patients having nail avulsions. The time for full re-growth ranged from 5 to 10 months. CONCLUSIONS: Total nail avulsion is an effective management option for patients whose diagnosis of onychomycosis was doubtful, and constitutes a treatment armamentarium especially for patients with single or oligo-onychomycosis.

Pressure-drop viscosity measurements for gamma-Al2O nanoparticles in water and PG-water mixtures (nanofluids).

Nanofluids have attracted wide attention because of their promising thermal applications. Compared with the base fluid, numerous experiments have generally indicated increases in effective thermal conductivity and convective heat transfer coefficient for suspensions having only a small amount of nanoparticles. It is also known that with the presence of nanoparticles, the viscosity of a nanofluid is greater than its base fluid and deviates from Einstein's classical prediction. However, only a few groups have reported nanofluid viscosity results to date. Therefore, relative viscosity data for gamma-Al2O3 nanoparticles in DI-water and propylene glycol/H2O mixtures are presented here based on pressure drop measurements of flowing nanofluids. Results indicate that with constant wall heat flux, the relative viscosities of nanofluid decrease with increasing volume flow rate. The results also show, based on Brenner's model, that the nanofluid viscosity can be explained in part by the aspect ratio of the aggregates.

The prevalence of microalbuminuria among patients with type II diabetes mellitus in a primary care setting: cross-sectional study.

OBJECTIVES: To determine the prevalence of microalbuminuria among patients with type II diabetes mellitus in a primary care setting, and to study the association between various risk factors and the presence of microalbuminuria. DESIGN: Cross-sectional community-based study. SETTING: Four primary care clinics, Hong Kong. PATIENTS: All patients with type II diabetes mellitus who regularly attended the clinics between May 2002 and March 2003. MAIN OUTCOME MEASURES: Patients' demographic data, the proportion with microalbuminuria (measured using a spot urine test), and the association between this condition and risk factors for diabetic nephropathy (via correlation and multivariable logistic regression analysis). RESULTS: The mean age of the 1161 patients in the sample population was 58.0 years. The mean duration of diabetes mellitus was 5.7 years, and the mean level of glycated haemoglobin was 7.4%. A total of 13.4% of the patients had microalbuminuria. Having the condition was significantly associated with advanced age, female sex, poor glycaemic control, and coexisting hypertension in both correlation and regression analyses. No significant association with ever smoking was found. CONCLUSION: Early screening for incipient diabetic nephropathy and aggressive management of modifiable risk factors in a primary care setting may be important in optimising the renal outcome of patients with type II diabetes mellitus.

Do men have a higher case fatality rate of severe acute respiratory syndrome than women do?

Severe acute respiratory syndrome (SARS) has been reported in 30 countries and regions, with a cumulative total of 8,099 probable cases and 774 deaths as of July 31, 2003, according to the World Health Organization. In Hong Kong, People's Republic of China, 1,755 SARS cases and 299 deaths had occurred as of September 22, 2003. The authors analyzed data from the Department of Health, Hong Kong SAR. The data series includes details regarding sex, age, and chronic disease history. Using data from early March to September 22, 2003, the authors found that males had a significantly (p < 0.0001) higher case fatality rate than females did, 21.9% versus 13.2%; the relative risk was 1.66 (95% confidence interval (CI): 1.35, 2.05), and it was 1.62 (95% CI: 1.21, 2.16) after adjustment for age. Subgroup analysis was conducted by excluding health care workers (n = 386) from the analysis. The overall crude relative risk of mortality was 1.41 (95% CI: 1.15, 1.74), and the adjusted relative risk was 1.48 (95% CI: 1.10, 2.00). Thus, among SARS patients, males may be more severely affected by the disease than females are. This finding could be related to a nonuniform case definition of SARS disease, a different treatment regimen, a past smoking history, work-environment factors, or gender-specific immune-defense factors, for instance.

Retrospective analysis of 196 patients with suspected pulmonary thromboembolism.

BACKGROUND: There has not been a statistical analysis reporting on patients with pulmonary thromboembolism (PE) in Taiwan. To identify the most common manifestations in patients with PE in Taiwan, we conducted this retrospective study. METHODS: We collected and analyzed the medical records of 196 patients (53 men, mean age, 60 years; 143 women, mean age, 46 years) whose clinical symptoms and signs, chest radiographs and lung scan findings were suggestive of PE. Conventional chest radiography was performed 24 hours to 48 hours before lung scans. The radiopharmaceuticals used in lung scans were 133Xenon for ventilation studies and 99mTechnetium macroaggregated albumin for perfusion studies. All patients had received anticoagulant therapy (heparin and coumadin) to improve PE. RESULTS: The two most common symptoms and signs of PE in the 196 patients were dyspnea (172/196, 87.8%) and tachypnea (176/196, 89.8%). The most common abnormal finding on chest radiography was increased lung markings, which were present in about 36.7% (72/196) of patients. Lung scans showed two or more, large, mismatched, segmental defects (high probability of PE) in both lungs in about 88.8% (174/196) of patients. After anticoagulant therapy, PE, as seen on lung scans, was resolved within four weeks in approximately 93.9% (184/196) of patients. CONCLUSIONS: In Taiwan, a woman aged 40 to 50 years, with no prior history of other medical problems or surgical procedures, with sudden onset of dyspnea or tachypnea, normal or increased lung markings on chest radiography and two or more large mismatched segmental defects on lung scans, is at greatest risk of developing PE. Most patients with PE showed a good response to anticoagulant therapy within four weeks.

Characterization of a novel allergen, a major IgE-binding protein from Aspergillus flavus, as an alkaline serine protease.

Aspergillus species of fungi have been known to be one of the most prevalent aeroallergens. One important A. flavus allergen (Asp fl 1) was identified by means of immunoblotting with a serum pool of allergic patients on a two-dimensional electrophoretic gel. The cDNA coding for Asp fl 1 was cloned and sequenced. The clone encodes a full-length protein of 403 amino acid precursors of 42 kDa. After cleavage of a putative signal peptide of 21 amino acids and a prepeptide of 100 amino acids, a mature protein of 282 amino acids was obtained with a molecular mass of 33 kDa and a pI of 6.3. A degree of identity was found in a range of 27 to 84% among related allergens derived from bacteria allergen subtilisin, mold allergen Pen c 1, and virulence factor of A. fumigatus. Recombinant Asp fl 1 (rAsp fl 1) was cloned into vector pQE-30 and expressed in E. coli M15 as a histidine-tag fusion protein and purified to homogeneity. The IgE binding capacity of rAsp fl 1 was tested by immunoblotting using a serum pool of Aspergillus-allergic patients. Recombinant allergen cross-reacted strongly with IgE specific for natural Asp fl 1 and Pen c 1, indicating that common IgE epitopes may exist between allergens of A. flavus and P. citrinum.

Expression of the crustacean hyperglycaemic hormones and the gonad-inhibiting hormone during the reproductive cycle of the female American lobster Homarus americanus.

Crustacean reproduction is regulated by a complex chain of hormonal interactions in which the crustacean hyperglycaemic hormones A and B (CHH-A and CHH-B) and the gonad-inhibiting hormone (GIH) play a primary role. These neurohormones are produced in the same neuroendocrine cells of the X-organ sinus gland complex, situated in the eyestalks of the American lobster, Homarus americanus. In order to obtain more information on the synthesis, storage, release and function of these three neuropeptides during the reproductive cycle, we studied the levels of their mRNAs in the X-organ, their peptide storage in the sinus gland and their concentration in the haemolymph at different stages of the female reproductive cycle. A high CHH-A mRNA level was found only in the previtellogenic stage, while elevated mRNA levels were determined for CHH-B in the mature as well as the previtellogenic stage. High CHH storage levels in the sinus gland were found during previtellogenesis. The total amount of CHH (CHH-A plus -B) in the haemolymph was significantly higher during maturation. A low level of GIH mRNA in the X-organ and a low amount of the GIH I isoform in the sinus gland were found only in the immature stage. In contrast, GIH haemolymph levels were high during the immature and previtellogenic stages. We conclude that CHH-A and -B are involved in triggering the onset of vitellogenesis and that CHH-B in particular is responsible for stimulating oocyte maturation before spawning, while GIH prevents the start of vitellogenesis in the ovary. Moreover, our results show that the balance between the haemolymph levels of the CHHs and GIH may tune the synchronization of reproduction and molting during the biannual reproductive cycle of the American lobster.

Prediction of the severity of haemolytic disease of the newborn. Quantitative IgG anti-D subclass determinations explain the correlation with functional assay results.

Sera containing anti-D, taken from 44 RhD-negative women with RhD-positive infants, were tested in antibody-dependent cellular cytotoxicity (ADCC) and monocyte monolayer assays (MMA) which used similar target and effector cell populations. In addition, the anti-D concentration was measured in the Auto Analyzer and the number of IgG1 and IgG3 anti-D molecules bound to the target red cells was measured by flow cytometry. The results of the functional assays and Auto Analyzer quantitation were examined for correlation with IgG subclass quantitation and all results were compared for their ability to predict the severity of haemolytic disease of the newborn (HDN). ADCC correctly predicted HDN in 39/44 (88.6%) cases, Auto Analyzer quantitation in 35/44 (79.5%) and the MMA in 32/44 (72.7%). For all three assays, the number of correct predictions was highest when the maternal serum contained both IgG1 and IgG3 anti-D. ADCC activity and HDN were correlated with the number of cell-bound IgG1 molecules (r > or = 0.58), but MMA activity was most closely correlated with the number of cell-bound IgG3 molecules (r = 0.68). Hence the superior predictive value of ADCC is due to its ability to reflect the IgG1 component of maternal anti-D, which has a better correlation than IgG3 anti-D with the severity of HDN.