Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma.

Flow cytometry has become a highly valuable method to monitor minimal residual disease (MRD) and evaluate the depth of complete response (CR) in bone marrow (BM) of multiple myeloma (MM) after therapy. However, current flow-MRD has lower sensitivity than molecular methods and lacks standardization. Here we report on a novel next generation flow (NGF) approach for highly sensitive and standardized MRD detection in MM. An optimized 2-tube 8-color antibody panel was constructed in five cycles of design-evaluation-redesign. In addition, a bulk-lysis procedure was established for acquisition of ⩾107 cells/sample, and novel software tools were constructed for automatic plasma cell gating. Multicenter evaluation of 110 follow-up BM from MM patients in very good partial response (VGPR) or CR showed a higher sensitivity for NGF-MRD vs conventional 8-color flow-MRD -MRD-positive rate of 47 vs 34% (P=0.003)-. Thus, 25% of patients classified as MRD-negative by conventional 8-color flow were MRD-positive by NGF, translating into a significantly longer progression-free survival for MRD-negative vs MRD-positive CR patients by NGF (75% progression-free survival not reached vs 7 months; P=0.02). This study establishes EuroFlow-based NGF as a highly sensitive, fully standardized approach for MRD detection in MM which overcomes the major limitations of conventional flow-MRD methods and is ready for implementation in routine diagnostics.

First results of a simultaneous measurement of tritium and (14)C in an ultra-low-background proportional counter for environmental sources of methane.

Simultaneous measurement of tritium and (14)C would provide an added tool for tracing organic compounds through environmental systems and is possible via beta energy spectroscopy of sample-derived methane in internal-source gas proportional counters. Since the mid-1960's atmospheric tritium and (14)C have fallen dramatically as the isotopic injections from aboveground nuclear testing have been diluted into the ocean and biosphere. In this work, the feasibility of simultaneous tritium and (14)C measurements via proportional counters is revisited in light of significant changes in both the atmospheric and biosphere isotopics and the development of new ultra-low-background gas proportional counting capabilities for small samples (roughly 50 cc methane). A Geant4 Monte Carlo model of a Pacific Northwest National Laboratory (PNNL) proportional counter response to tritium and (14)C is used to analyze small samples of two different methane sources to illustrate the range of applicability of contemporary simultaneous measurements and their limitations. Because the two methane sources examined were not sample size limited, we could compare the small-sample measurements performed at PNNL with analysis of larger samples performed at a commercial laboratory. These first results show that the dual-isotope simultaneous measurement is well matched for methane samples that are atmospheric or have an elevated source of tritium (i.e. landfill gas). However, for samples with low/modern tritium isotopics (rainwater), commercial separation and counting is a better fit.

Persistence of influenza vaccine-induced antibodies in lung transplant patients between seasons.

BACKGROUND: Immunization policy-making bodies advised against immunizing too early before the influenza season because vaccine-specific antibody may wane before the end of the influenza season. Lung transplant patients are included in the group of high-risk patients for whom this recommendation had been made. We hypothesized that immunosuppressed lung transplant patients would maintain protective concentrations of influenza antigen-specific antibodies between seasons. METHODS: As part of a planned 5-year study of influenza vaccine responses in lung transplant patients, we measured influenza antibody concentrations by hemagglutination inhibition assay before influenza immunization annually. The fraction of lung transplant patients who maintained seroprotective levels (≥ 40 hemagglutination units) approximately 11 months from last season immunization was calculated. Antibody concentrations and response rates in lung transplant patients were compared with healthy individuals and those waiting for lung transplantation. RESULTS: The majority of lung transplant patients maintained seroprotective influenza antigen-specific antibody concentrations for approximately 11 months after immunization. Seroprotection rates varied greatly with influenza antigens (healthy 68-100%, pretransplant 44-100%, transplant 64-100%), and were similar when groups were compared. More than 70% of lung transplant patients maintained seroprotective antibody concentrations to 10 of 11 vaccine antigens. CONCLUSION: Seroprotective influenza antibody concentrations are maintained at very high rates among immunosuppressed lung transplant patients and depend more on the vaccine virus than the immunostatus of the vaccine recipient. Early seasonal influenza immunization of lung transplant patients is appropriate.

Multiple organ failure in trauma patients.

SUMMARY: BACKGROUND As care of the critically ill patient has improved and definitions of organ failure have changed, it has been observed that the incidence of organ failure and the mortality associated with organ failure appear to be decreasing. In addition, many early studies included large heterogeneous populations of both medical and surgical patients that may have influenced the incidence and outcome of organ failure. The purpose of this study is to establish the current incidence and mortality of organ failure in a homogenous population of critically ill trauma patients. METHODS All trauma patients admitted to the intensive care unit (ICU) at an urban Level I trauma center were prospectively studied. Patients were evaluated for the presence of organ failure using definitions proposed by Knaus and by Fry. Newer definitions of organ failure incorporating organ dysfunction and severity-of-illness scores were also obtained in all patients in an attempt to predict outcome. These included lung injury scores (acute respiratory distress syndrome scores), Acute Physiology and Chronic Health Evaluation (APACHE) II and III scores, Injury Severity Score (ISS), and multiple organ dysfunction scores. Primary outcomes assessed were death and the occurrence of organ failure by the various definitions. RESULTS Eight hundred sixty-nine trauma patients were admitted to the ICU and survived longer than 48 hours. Mean APACHE II and APACHE III scores at admission to the ICU and ISS were 12.2 +/- 22, 30.5 +/- 22.7, and 19 +/- 10, respectively. Single organ failure (SOF) occurred in 163 patients (18.7%) and multiple organ failure occurred in 44 patients (5.1%). All SOF was caused by respiratory failure. Respiratory failure occurred first in the majority of patients with multiple organ failure. Mortality was 4.3% with one organ system failure, 32% with two, 67% with three, and 90% when four organ systems failed. None of the patients with SOF died secondary to respiratory failure. Multiple stepwise regression analysis was performed to determine which of the following risk factors are associated with the occurrence of organ failure: mechanism of injury, lactate at 24 hours, ISS, APACHE II, APACHE III, acute respiratory distress syndrome score at admission, multiple organ dysfunction score at admission and total blood products transfused in 24 hours. Of these factors, APACHE III, lactate at 24 hours, and total blood products transfused in 24 hours were associated with the occurrence of organ failure. CONCLUSION The overall incidence of organ failure in a homogeneous trauma population appears to be lower than that reported in studies performed in heterogeneous patient populations in the 1980s. Mortality for SOF is low and appears to be related primarily to the patient's underlying injuries and not to organ failure. Mortality for two or three organ system failures is lower than reported 15 to 20 years ago. Mortality for patients with four or more organ system failures remains high, approaching 100%.

Comparisons among children's responses to the Hand Test by grade, race, sex, and social class.

Four hundred sixty-four children in grades K-8 of an urban school were tested in order to develop norms, check for the presence of developmental trends, and compare the performance of different demographic groups on the Hand Test. Comparisons across grade, race, sex, and socioeconomic class indicated that separate norms for grades, but single norms for race, sex, and socioeconomic levels are appropriate for elementary school children. Discrepancies in the developmental trends raised concern about the validity of indices of psychopathology especially as they apply to children in grades K-8.

Familial lobular glomerulopathy.

A family with an unusual lobular glomerulopathy is described. Renal tissue from three males and one female in two successive generations was available for review. The glomerulopathy was characterized by a marked lobular accentuation with only a modest increase in mesangial cellularity. Immunofluorescence in two patients showed focal or diffuse staining with immunoglobulins G, A, M, and C3 in the mesangium and along the glomerular capillary basement membranes. Ultrastructural study showed amorphous granular subendothelial material distending capillary loops and mesangial regions. This material accounted for the pronounced lobular accentuation. The patients in this family presented with proteinuria, hematuria, and hypertension. Three of the four patients have sustained cerebral vascular accidents and two have died. This family is compared with a previously reported family that showed similar glomerular pathology.

Interscorer reliability for the Hand Test administered to children.

The utility of the Hand Test as a quick, reliable measure of 100 children's personalities was assessed. The interscorer reliability of the Hand Test was estimated by both intraclass correlations and the Kappa coefficient for 100 children. Following training, satisfactory intraclass correlations were obtained for the Quantitative scores (20 of 22 above .70) and Qualitative scores (12 of 27 above .70) Kappa coefficients were generally lower. Scorers' memory overload and low response frequency are discussed as possible bases for the low reliabilities of Qualitative scores. Although the Hand Test reliability for Quantitative scores is consistent with those of other projective tests, consideration should be given to the modification of the directions of administration for young children and clarification of scoring rules.

Stress fractures in pregnancy.

Because of jogging and aerobics, more stress fractures are occurring in women today. That they can occur unrelated to these activities in pregnant women is not quite so well known. Two cases of stress fracture of the pubic bone sustained at delivery are presented, along with discussion of the possible causes and simple treatment of such fractures.

Hypocholesterolemic effect of khellin and khelloside in female cynomolgus monkeys.

Khellin and khelloside (khellol glucoside) were examined in female cynomolgus monkeys to substantiate their ability to favorably modify serum lipoprotein cholesterol. Clinical chemistry parameters were also measured to obtain information indicative of possible drug toxicity. Both drugs were evaluated in two week multiple-dose studies and after a single oral dose. After two weeks at 20 mg/kg per day, khellin and khelloside significantly lowered low density lipoprotein cholesterol (LDL-C) by 87% and 73%, high density lipoprotein cholesterol (HDL-C) by 41% and 23%, and total-C by 55% and 44%, respectively. Very low density lipoprotein cholesterol (VLDL-C) and triglycerides (TG) were not changed. No apparent toxicity was observed as clinical chemistry parameters and body weights were not different compared to control values. Similar results were observed with lower doses of khellin and khelloside. Khellin at 5 mg/kg per day reduced LDL-C by 50%, HDL-C by 15%, and total-C by 30%, while khellol glucoside at 10 mg/kg per day lowered LDL-C by 46%, HDL-C by 20%, and total-C by 31%. Neither drug produced significant changes in VLDL-C, TG, body weights, or clinical chemistry variables. A 2 mg/kg per day dose of khellin also had no observable effect in this study. Single oral doses (20 mg/kg) of khellin and khelloside caused modulation of LDL-C (-32% and -30%) and total-C (-18% and -15%). Visual observation of the monkeys during this study revealed that khellin caused emesis in 9/9 animals, while khelloside and control had no emetic effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Amyloidosis in adult onset Still's disease.

A patient with adult onset of Still's disease who developed secondary amyloidosis is described. This is the first report of the association of these 2 disease states. The association of amyloidosis and other rheumatic disorders is discussed.

Chronic endometritis: morphologic and clinical observations.

The histologic and clinical manifestations of chronic endometritis were reviewed in 99 women. The morphologic features found to be of value in diagnosing this condition were superficial stromal edema, increased stromal density, and pleomorphic stromal inflammatory infiltrate dominated by lymphocytes in the absence of premenstrual changes or any other significant pathologic endometrial lesions. When these changes were present, a plasma cell infiltrate was invariably found. Clinically, the major presenting complaint was vaginal bleeding in 94% of the patients. No correlation was found between the presenting clinical complaint and either the extent of the lesion or the number of plasma cells in the leukocyte infiltrate. On follow-up, the lesion appeared to be eradicated by biopsy or curettage in approximately 80% of the patients. The major predisposing conditions were found to be intrauterine leiomyomas and a recent endometrial biopsy or curettage. The limitations of the plasma cells criterion for recognition of the lesion are discussed.

Idiopathic arterial calcification of infancy: a clinicopathologic study.

The clinical and pathologic features of a group of 62 infants dying of idiopathic calcific arterial disease were reviewed. The disease most commonly occurs in infants less than 6 months of age. Pathologically, it is characterized by calcific deposits along the internal elastic membrane of arteries accompanied by fibrous thickening of the intima which causes luminal narrowing. The arterial lesions are widespread but the resultant luminal narrowing invariably promotes myocardial ischemia, causing the infants' deaths. A definite tendency of the disease to occur in siblings has been noted, but additional patterns of inheritance are not yet apparent. Clinical diagnosis is feasible with radiologic study of arteries of the head, neck, and extremities. There is a similarity of idiopathic calcific arterial disease of infancy to the arterial lesions of metastatic calcification in severe renal disease, calcific arterial lesions noted in conjunction with certain cardiovascular anomalies, and hypervitaminosis D. Certain experimental situations and toxic states can also produce calcific and proliferative vascular lesions.