bluuebunny/crossref_metadata_2025 huggingface dataset:
PRODUCTION OF SECONDARY METABOLITES BY FILAMENTOUS TROPICAL MARINE CYANOBACTERIA: ECOLOGICAL FUNCTIONS OF THE COMPOUNDS
Authors: Paul Dale G. Nagle and Valerie J.
Published: March 2003

Cyanobacterial blooms are becoming more common in many reef habitats. The broadly acting feeding deterrent compound ypaoamide, produced by a mixed cyanobacterial assemblage, has been linked to bloom formation and mass fish die‐offs (Siganus argenteus and Siganus spinus) in Guam. Specific metabolites produced by Lyngbya majuscula Gomont act as both feeding attractants to the specialist herbivore Stylocheilus longicauda, and as effective feeding deterrents to generalist fishes. Two‐dimensional TLC (2D‐TLC) analysis of cyanobacterial crude extracts was used to select chemically distinct populations (chemotypes) of bloom‐forming filamentous cyanobacteria for chemical and ecological evaluation. Crude extracts produced by different species, chemotypes, and chemically distinct Micronesian marine cyanobacterial assemblages deter feeding activity of generalist reef herbivores. The ecological function of cyanobacterial secondary metabolites, especially as related to diversity of compound production and the relationship of metabolite production to bloom formation is discussed.
VALIDATION OF SOME CURRENTLY USED INVALID NAMES OF DINOFLAGELLATES
Authors: Trench Robert
Published: March 2003

Holotypes are designated to validate the names Symbiodinium goreaui, Symbiodinium kawagutii, Symbiodinium pilosum, Scrippsiella velellae, and Gloeodinium viscum (Dinophyceae).
Free proline levels in ulva (chlorophyta) in response to hypersalinity: elevated nacl in seawater versus concentrated seawater (Note)
Authors: Liu Chia‐Hsiung , Shih Ming‐Chiao , Lee Tse‐Min
Published: March 2003

Changes in free proline concentrations following exposure to elevated NaCl in 35 psu artificial seawater (ASW) (an imbalance in ion ratio) or concentrated ASW (constant ion ratio) were investigated in the intertidal green alga Ulva fasciata Delile. When exposed to 55 psu achieved by adding NaCl to 35 psu ASW, free proline concentrations increased at hour 3 and reached a plateau after 12 h, whereas free proline concentrations in response to 55 psu achieved by concentrating ASW were constant during the treatment period. It is likely that an imbalance in ion ratio is a cue for the induction of a moderate accumulation of free proline in U. fasciata.
GENETIC VARIATION IN APHANIZOMENON (CYANOBACTERIA) COLONIES FROM THE BALTIC SEA AND NORTH AMERICA
Authors: Barker Gary L. A. , Konopka Allan , Handley Barbara A. , Hayes Paul K.
Published: March 2003

Aphanizomenon Morren is an important member of the cyanobacterial community in the Baltic Sea, but studies of this genus have been hampered by the difficulty of growing it in laboratory culture. PCR amplification of DNA from colonies picked directly from water samples has circumvented this problem and made it possible to carry out an analysis of genetic diversity within the Baltic Sea and in two small North American lakes separated by just a few kilometers. The nucleotide sequence of the phycocyanin intergenic spacer and partial flanking coding regions of cpcB and cpcA was determined for 32 colonies of Aphanizomenon, 26 from the Baltic Sea, and 6 from the North American lakes. No variation was detected among the 26 Baltic Sea colonies, but two alleles, differing at 19 nucleotide positions (5.4%), were found in the North American lake colonies. Surprisingly, the two North American types were less closely related to each other than to the Baltic Sea genotype. The Baltic Sea Aphanizomenon is clearly distinct from A. flos‐aquae at both the cpcB–cpcA and 16S rDNA loci, which lends phylogenetic support to their tentative separation based on ultrastructural analysis. We conclude that although there is significant genetic diversity in the genus Aphanizomenon, the population in the Baltic Sea is, in contrast to the Nodularia population, genetically homogeneous.
EVOLUTION OF AN ARTIFICIAL SEAWATER MEDIUM: IMPROVEMENTS IN ENRICHED SEAWATER, ARTIFICIAL WATER OVER THE LAST TWO DECADES
Authors: Berges John A. , Franklin Daniel J. , Harrison Paul J.
Published: March 2003

Although most phycologists use natural seawater for culturing marine species, artificial media continue to play important roles in overcoming problems of supply and seasonal variability in the quality of natural seawater and also for experiments involving manipulation of micro‐ and macronutrients. Several artificial media have been developed over the last 90 years; enriched seawater, artificial water (ESAW) is among the more popular recipes. ESAW has the advantage of an ionic balance that is somewhat closer to that of normal seawater. The original paper compared the growth of 83 strains of microalgae in natural seawater (ESNW) versus ESAW and determined that 23% grew more poorly in the artificial water. Since 1980, however, the composition of ESAW, as used by the original authors, has changed considerably. In particular, the added forms of phosphate, iron, and silicate have been changed and the trace metal mixture has been altered to include nickel, molybdenum, and selenium. We tested whether these changes improved the ability of the artificial medium to grow previously difficult to grow phytoplankton species. To test this, we selected eight species that had been shown to grow better in ESNW than in ESAW and compared their growth again, using the currently used recipe with all the above modifications. For all but one species (Apedinella spinifera), growth rate and final yield was no different between the media but in one case (Emiliania huxleyi) was slightly higher in ESAW. No differences in cell morphology or volume were found in any case. We conclude that changes to the enrichment portion of the recipe have significantly improved this artificial seawater medium and that it can be used to grow an even wider range of coastal and open ocean species.
INFECTION OF PROROCENTRUM MINIMUM (DINOPHYCEAE) BY THE PARASITE AMOEBOPHRYA SP. (DINOFLAGELLEA)
Authors: Maranda Lucie
Published: March 2003

In early summer 1998, the parasitic dinoflagellate Amoebophrya sp. Koeppen infected a population of the photosynthetic dinoflagellate Prorocentrum minimum (Pavillard) Schiller in a temperate shallow embayment. This is the first record of such an infection in P. minimum. The parasite was observed live, both within its deformed host as a trophont and outside its host, either as a swimming mass of flagellated cells, the vermiform, or as individual sporonts. Except for the shape of the vermiform, stages of development were in accordance with previous descriptions of Amoebophrya parasitizing photosynthetic dinoflagellates. Planktonic grazers fed on the sporonts. Further studies are needed to decipher the influence of parasitism on the regulation of host abundance.
GROWTH RATE RELATIONSHIPS TO PHYSIOLOGICAL INDICES OF NUTRIENT STATUS IN MARINE DIATOMS
Authors: Clark Darren R.
Published: March 2003

The growth of two species of marine diatom, Thalassiosira weissflogii (Grunow) and Thalassiosira pseudonana (Hustedt), was followed in batch cultures at four concentrations of dissolved inorganic carbon from N‐ and C‐replete lag phase into N‐ and/or C‐deplete stationary phase. Results describe the relationship between carbon‐specific growth rate (μC) and chl a:carbon (chl a:C) and glutamine:glutamate (gln:glu) ratios with changes in the cells' nutritional status (N:C), during the utilization of either NO3− or NH4+. The use of the gln:glu ratio as an index of N:C requires further clarification. For both species and N sources, N stress resulted in a decrease in μC, chl a:C, and N:C relative to μCmax values, whereas C stress resulted in a decrease in μC and an increase in chl a:C and N:C relative to μCmax values. Both species attained a chl a:C ratio of approximately 15 μg·g−1 at μCmax using either N source. However, this value was not necessarily an indicator of maximal growth rate. NC colimitation resulted in decreased μC to values less than 20% of μCmax with only minor changes in chl a:C and N:C relative to μCmax values. Chl a:C results suggest a similarity between the light stress and C stress responses of marine diatoms. The potential for C stress in the marine environment needs to be addressed.
DNA DAMAGE AND PHOTOSYNTHETIC PERFORMANCE IN THE ANTARCTIC TERRESTRIAL ALGA PRASIOLA CRISPA SSP. ANTARCTICA (CHLOROPHYTA) UNDER MANIPULATED UV‐B RADIATION
Authors: Lud Daniela , Buma Anita G.J. , Van De Poll Willem , Moerdijk Tanja C.W. , Huiskes Ad H.L.
Published: March 2003

The effect of reduced, natural ambient, and enhanced UV‐B radiation (UVBR) on photosynthesis and DNA damage in the Antarctic terrestrial alga Prasiola crispa ssp. antarctica (Kützing) Knebel was investigated in two field experiments. Samples of P. crispa were collected underneath snow cover and exposed outside to reduced and natural UVBR in the austral spring. In a second experiment at the end of the austral summer, samples were exposed to ambient and enhanced UVBR. PSII efficiency, net photosynthetic rate (NP), dark respiration rate (DR), UV‐absorbing pigments, and cyclobutyl pyrimidine dimer (CPD) formation were measured during the experiments.

In October 1998, a spring midday maximum of 2.0 W·m−2 of UVBR did not significantly affect effective quantum yield (ΔF/Fm′), and a reduction in the ratio of variable to maximal fluorescence (Fv/Fm) in the late afternoon was transient. Exposure to natural ambient UVBR in October increased CPD values significantly. Midday maxima of UVBR during the experiments in October and January were comparable, but Setlow‐DNA‐weighted UVBR was more than 50% lower in January than in October. In January, 0.5 W·m−2 additional UVBR during 10 h did not have a negative effect on ΔF/Fm′. The reduction in Fv/Fm was not significant. NP and DR were not affected by supplementation of UVBR. Although photosynthetic activity remained largely unaffected by UVBR treatment, DNA damage was shown to be a sensitive parameter to monitor UVBR effects. Supplementation of additional UVBR did significantly enhance the amounts of CPD in exposed samples and repair took place overnight. It is concluded that PSII and whole‐chain photosynthesis of P. crispa is well adapted to ambient and enhanced levels of UVBR but that CPD formation is more sensitive to UVBR than to photosynthesis.
LIGHT BOUNDARIES AND THE COUPLED EFFECTS OF SURFACE HYDROPHOBICITY AND LIGHT ON SPORE SETTLEMENT IN THE BROWN ALGAHINCKSIA IRREGULARIS(PHAEOPHYCEAE)1
Authors: Greer Stephen P. , Amsler Charles D.
Published: March 2003

We examined the effects of light and surface hydrophobicity individually and in tandem onHincksia irregularis(Kützing) Amsler spore settlement.Hincksia irregularisspores were determined to be negatively phototactic by the use of computer‐assisted motion analysis. Spore settlement was significantly influenced by surface hydrophobicity and by light, individually and in tandem. Experiments conducted using culture plates modified to reduce well edge artifacts revealed significantly higher settlement on hydrophobic surfaces and in dark environments when compared with negatively charged surfaces and lighted environments. Experimental light/dark boundaries elicited distinct spore settlement responses, with spores displaying dissimilar settlement patterns on plates with different surface hydrophobicities. The results of this study indicateH. irregularisspores possess the capacity for complex responses to their environment. These complex responses may influence dispersal and aid spores in the detection of suitable settlement locations in marine microenvironments.
ENVIRONMENTAL FACTORS CONTROLLING PHYTOPLANKTON PROCESSES IN THE SOUTHERN OCEAN1
Authors: Boyd Philip W.
Published: March 2003

The factors controlling the distribution of phytoplankton stocks, species composition, and their physiological status in the Southern Ocean are reviewed. In the last decade, the key data sources have been observational and experimental. Together, they provide a framework to understand the complex temporal and spatial patterns of environmental control within the distinct basins and ecological provinces. High resolution remotely sensed observational data have overcome the issue of geographical remoteness. Furthermore, by exploiting seasonal and spatial differences in algal distributions, observational data have enabled the cross‐correlation of such trends with patterns in other environmental properties. Perturbation experiments have offered a mechanistic understanding to help interpret observational data by altering environmental properties under carefully controlled conditions. A consistent set of trends, on the modes of environmental control of phytoplankton processes, is now emerging across the different basins and provinces. The key determinants are light, iron, and silicic acid supply (top‐down control was not considered). However, their interplay in time and space (i.e. simultaneous limitation of phytoplankton processes) is less clear, requires further study, and is discussed. Future challenges include the need to understand better the mode(s) of environmental control on key algal functional groups via more taxon‐ and species‐specific studies. The initiation of more time‐series moorings with “smart” bio‐optical and sampling sensors are needed to define the seasonal distributions of algal taxa. Moreover, new perturbation experiments are required to investigate the influence on phytoplankton processes of projected climate‐mediated alteration of mixed layer depth and nutrient supply as widely predicted by modelers.
New Insights in the Systematics of the Phyllophoraceae (Gigartinales, Rhodophyta)
Authors: Fredericq S. , Lopez‐Bautista J. M. , Moe R. L.
Published: March 2003

The Phyllophoraceae, consisting of about 100 species worldwide, stands out in exhibiting a wide spectrum of unique life history types that makes it unusually interesting for assessing the phylogenetic importance of reproductive traits relative to classification criteria. Type of life history and position of the reproductive structures on the thallus have traditionally formed the basis for separating 11 genera in the Phyllophoraceae; however, phylogenetic analyses inferred from three sets of DNA sequences [chloroplast‐encoded rbcL, nuclear large‐subunit ribosomal RNA gene (LSU rDNA), and internal transcribed spacer regions (ITS) of nuclear ribosomal DNA], instead indicate a lack of correlation between type of life history and phylogenetic relationships among the established taxa. This lack of correlation dramatically challenges all of the traditional taxonomy, and we will present a revised classification for the family that downplays life‐history features. The study will answer the question which morphological features can be used as meaningful indicators of phylogenetic relationships in the Phyllophoraceae. The results will also be addressed in light of global biogeographic hypotheses for the family.
Calcification and Decalcification During Epithallial Cell Turnover in Coralline Red Algae
Authors: Pueschel C. , Wegeberg S.
Published: March 2003

Epithallial cells of the coralline red algae are characterized by unusual structural specialization, which include deep invaginations of the distal cell surface, and by unique development, which culminates in senescence, shedding, and replacement of the cells. Electron microscopic study of epithelial cell differentiation in morphologically and taxonomically disparate species suggests that the unusual features of epithelial cell structure and development stem from the fact that these dynamics occur within a calcified matrix. Distal wall ingrowths begin to form on the initial cells, cells whose cleavage eventually gives rise distally to new epithelial cells. After the distal wall ingrowths form, the overlying crosswall becomes rich in organic material. For this organic wall material to be deposited into the existing crosswall, the wall must first be decalcified; therefore, the presence of abundant organic material in the crosswall provides a marker of localized decalcification. We propose that the location and time of origin of distal wall ingrowths indicate a connection between the ingrowths and two coordinated processes: localized secretion of wall material, and decalcification of the overlying cell wall in preparation for the movement of the young epithelial cell into a new location relative to the surrounding calcified matrix. The large plasmalemmal surface area associated with the distal wall ingrowths allows for a greater abundance of membrane‐associated components, such as proton pumps, that could drive localized cell wall decalcification.
Low Relative Skeletal Muscle Mass (Sarcopenia) in Older Persons Is Associated with Functional Impairment and Physical Disability
Authors: Janssen Ian , Heymsfield Steven B. , Ross Robert
Published: March 2003

OBJECTIVES: To establish the prevalence of sarcopenia in older Americans and to test the hypothesis that sarcopenia is related to functional impairment and physical disability in older persons.

DESIGN: Cross‐sectional survey.

SETTING: Nationally representative cross‐sectional survey using data from the Third National Health and Nutrition Examination Survey (NHANES III).

PARTICIPANTS: Fourteen thousand eight hundred eighteen adult NHANES III participants aged 18 and older.

MEASUREMENTS: The presence of sarcopenia and the relationship between sarcopenia and functional impairment and disability were examined in 4,504 adults aged 60 and older. Skeletal muscle mass was estimated from bioimpedance analysis measurements and expressed as skeletal muscle mass index (SMI = skeletal muscle mass/body mass × 100). Subjects were considered to have a normal SMI if their SMI was greater than ‐one standard deviation above the sex‐specific mean for young adults (aged 18–39). Class I sarcopenia was considered present in subjects whose SMI was within ‐one to ‐two standard deviations of young adult values, and class II sarcopenia was present in subjects whose SMI was below ‐two standard deviations of young adult values.

RESULTS: The prevalence of class I and class II sarcopenia increased from the third to sixth decades but remained relatively constant thereafter. The prevalence of class I (59% vs 45%) and class II (10% vs 7%) sarcopenia was greater in the older (≥ 60 years) women than in the older men (P < .001). The likelihood of functional impairment and disability was approximately two times greater in the older men and three times greater in the older women with class II sarcopenia than in the older men and women with a normal SMI, respectively. Some of the associations between class II sarcopenia and functional impairment remained significant after adjustment for age, race, body mass index, health behaviors, and comorbidity.

CONCLUSIONS: Reduced relative skeletal muscle mass in older Americans is a common occurrence that is significantly and independently associated with functional impairment and disability, particularly in older women. These observations provide strong support for the prevailing view that sarcopenia may be an important and potentially reversible cause of morbidity and mortality in older persons.
Development and Testing of a Measure of Health‐Related Quality of Life for Men with Urinary Incontinence
Authors: Robinson Joanne P. , Shea Judy A.
Published: March 2003

OBJECTIVES: To describe the development and psychometric testing of male versions of the Urogenital Distress Inventory (UDI) and Incontinence Impact Questionnaire (IIQ).

DESIGN: Instrument development.

SETTING: Urology clinic at a large urban Veterans Affairs Medical Center.

PARTICIPANTS: Convenience sample of English‐speaking community‐dwelling male urology clinic patients who acknowledged concerns about urine leakage.

MEASUREMENTS: Content experts and male clinic patients evaluated items for clarity and fit. Revised instruments were administered to 153 participants. Data were analyzed to examine issues of feasibility of administration, internal consistency reliability, and validity.

RESULTS: Modal completion time for the Male Urogenital Distress Inventory (MUDI) (27 items) and Male Urinary Symptom Impact Questionnaire (MUSIQ) (32 items) was 20 minutes. Cronbach's coefficients were .89 for the MUDI and .95 for the MUSIQ. Total MUDI and MUSIQ scores were moderately correlated (r = .59, P < .001). Mean MUDI and MUSIQ scores varied significantly with self‐reported desire for socialization, urine leakage, and depression. Principal components analyses suggested the presence of seven factors accounting for 65.4% of the variance in the MUDI and six factors accounting for 72.9% of the variance in the MUSIQ.

CONCLUSIONS: It is feasible to use the MUDI and MUSIQ to measure health‐related quality of life in men with continence problems. The scores appeared to be reliable and valid in this racially and educationally diverse sample. Use of the MUDI and MUSIQ may provide more sensitive measurement of the specific effect of urinary incontinence and related symptoms on health‐related quality of life in men. Future research should determine reproducibility and responsivity and reexamine the construct validity of these instruments.
Identifying Postmenopausal Women at High Risk of Fracture in Populations: A Comparison of Three Strategies
Authors: Buist Diana S.M. , LaCroix Andrea Z. , Manfredonia David , Abbott Thomas
Published: March 2003

OBJECTIVES: To describe the prevalence of risk factors for women at high risk of fracture in a population‐based sample of postmenopausal women who were not using hormone replacement therapy (HRT), to demonstrate how the estimated prevalence of women at high risk of future fracture is affected by the different criteria used for classification, and to characterize the populations identified and missed by each of the criteria. A key study objective was to compare the proportion of postmenopausal women at high risk of fracture in a managed care population using several different definitions of who is at high risk.

DESIGN: The Osteoporosis Population‐based Risk Assessment study, a randomized trial of three screening strategies.

SETTING: Conducted at Group Health Cooperative in western Washington state.

PARTICIPANTS: Women aged 60 to 79 who had not used HRT for at least 12 months were chosen at random.

MEASUREMENTS: In one of the trial arms, 428 women had their bone mineral density (BMD) measured at the hip and spine (L1–L4) using dual energy x‐ray absorptiometry. Minimum t scores and z scores at all sites were used for classification. Risk factors for fractures were assessed at the time of the BMD scan.

RESULTS: Guidelines based on the Study of Osteoporotic Fractures classified 25.1% of the women as being at high risk of fracture, compared with 30.0% and 68.0% using World Health Organization (WHO) recommendations and National Osteoporosis Foundation guidelines, respectively. Classification based on low BMD alone (WHO) failed to include more than 50% of women who had already experienced a clinical fracture.

CONCLUSIONS: Prevalence of women at high risk of fracture not using HRT varies notably depending on the criteria used for identification. The criteria used to identify women to target for primary and secondary prevention of osteoporotic fractures has major implications for population‐based prevention strategies.
Profiling Nursing Homes Using Bayesian Hierarchical Modeling
Authors: Berlowitz Dan R. , Christiansen Cindy L. , Brandeis Gary H. , Ash Arlene S. , Kader Boris , Morris John N. , Moskowitz Mark A.
Published: March 2003

OBJECTIVES: New methods developed to improve the statistical basis of provider profiling may be particularly applicable to nursing homes. We examine the use of Bayesian hierarchical modeling in profiling nursing homes on their rate of pressure ulcer development.

DESIGN: Observational study using Minimum Data Set data from 1997 and 1998.

SETTING: A for‐profit nursing home chain.

PARTICIPANTS: Residents of 108 nursing homes who were without a pressure ulcer on an index assessment.

MEASUREMENTS: Nursing homes were compared on their performance on risk‐adjusted rates of pressure ulcer development calculated using standard statistical techniques and Bayesian hierarchical modeling.

RESULTS: Bayesian estimates of nursing home performance differed considerably from rates calculated using standard statistical techniques. The range of risk‐adjusted rates among nursing homes was 0% to 14.3% using standard methods and 1.0% to 4.8% using Bayesian analysis. Fifteen nursing homes were designated as outliers based on their z scores, and two were outliers using Bayesian modeling. Only one nursing home had greater than a 50% probability of having a true rate of ulcer development exceeding 4%.

CONCLUSIONS: Bayesian hierarchical modeling can be successfully applied to the problem of profiling nursing homes. Results obtained from Bayesian modeling are different from those obtained using standard statistical techniques. The continued evaluation and application of this new methodology in nursing homes may ensure that consumers and providers have the most accurate information regarding performance.
Physicians Vary in Approaches to the Clinical Management of Delirium
Authors: Carnes Molly , Howell Timothy , Rosenberg Marjorie , Francis Joseph , Hildebrand Christopher , Knuppel Jeffrey
Published: March 2003

OBJECTIVES: To ascertain the variation in strategies for managing delirium of physicians with expertise in geriatrics.

DESIGN: Cross‐sectional mail survey.

SETTING: United States.

PARTICIPANTS: A probability sample of physician members of the American Geriatrics Society.

MEASUREMENTS: Management choices presented in a two‐part case vignette of an older woman hospitalized with a hip fracture who develops mild and then severe delirium.

RESULTS: One hundred twenty‐two respondents (43%) selected the three answers constituting current “best practice,” 50 (18%) selected an unnecessary diagnostic test (brain imaging, lumbar puncture, or electroencephalogram), and 47 (17%) selected unnecessary pharmacologic therapy for mild delirium. For severe delirium, 270 (96%) selected pharmacological therapy, of whom 180 chose haloperidol alone, 55 chose lorazepam alone, 23 chose lorazepam in combination with haloperidol, and 12 wrote in another drug. Thirty percent of the respondents made any selection of lorazepam, alone or in combination with haloperidol, for mild or severe delirium. Sixty‐one percent of those selecting haloperidol for severe delirium chose a dose greater than that recommended for geriatric patients. Sex, date of graduation from medical school, clinical specialty, completion of a geriatric fellowship, or certification in geriatrics had no significant effect on responses.

CONCLUSIONS: The common selection of lorazepam to treat delirium is troubling because benzodiazepines themselves are implicated in delirium. Selection of an initial dose of haloperidol higher than that recommended for geriatric patients by more than half of the respondents is also of concern. There is a paucity of sound clinical evidence to guide the choice of pharmacological agents for treating delirium in older hospitalized patients.
Survival of Kn(a+)McC(a+) red cells in a patient with anti‐“Kna/McCa”
Authors: Ballas S.K. , Viggiano E. , Draper E.K.
Published: March 2003

The in vivo survival of Kn(a+)McC(a+) red cells in a patient with anti‐ “Kna/McCa” was studied using 51Cr‐labeled incompatible cells. The antibody was IgG4, demonstrable at 37 degrees C in the antiglobulin test, and did not bind complement. Survival of tagged cells was 82.2 percent after 24 hours. The patient was transfused with a total of 29 units of incompatible blood with no evidence of ill effects. The direct antiglobulin test became positive after transfusion and remained positive for 2 1/2 months of observation. The findings suggest that individuals with anti‐ “Kna/McCa” may be transfused with Kn(a+)McC(a+) red cells.
Territorial issues addressed through regionalization
Authors: Gaynor S.M. , Peetoom F.
Published: March 2003

Possibly no structural issue in blood banking is as problematic as that of service “territory.” The Regionalization Program of the American Blood Commission has worked with numerous regions in an effort to achieve a rational resolution of territorial issues. This paper discusses approaches utilized by these regions and results achieved. It provides a discussion of an alternate model that facilitates resolution of some problems. This information may be beneficial to regions that encounter similar problems.
Long‐term cryopreservation of red cells from patients with sickle cell disease
Authors: Castro O. L.
Published: March 2003

Twelve units of red cells (RBCs) from patients with sickle cell disease were glycerolized and frozen for periods of 1 to 3 years. The freeze‐ thaw hemolysis (mean 11.6%±2.1% SEM) and in vitro process recovery (mean 74.4%±3.7% SEM) were similar to those of sickle RBCs frozen for shorter periods of time. There was a weak positive correlation between duration of frozen storage and cell losses during deglycerolization. The adenosine triphosphate and 2,3‐ diphosphoglycerate levels of the thawed RBCs were within an acceptable range (mean 3.4 ± 0.2 μmol/g Hb and 18.0 ± 0.8 μmol/g Hb, respectively). Long‐term cryopreservation of sickle (Hb SS) RBCs opens the possibility of exploring autologous transfusion to treat sickle cell disease patients during anemic episodes that are not due to sickling.
Effects on immunity of multiple leukapheresis using a rapidly excreted analog of hydroxyethyl starch
Authors: Strauss R. G. , Goeken J. A. , Imig K. M. , Greazel C.
Published: March 2003

Immunologic studies were performed immediately before and after leukapheresis in five subjects undergoing four granulocyte donations (separated by 3‐ to 4‐week intervals) using continuous‐flow centrifugation leukapheresis with a rapidly excreted investigational analog of hydroxyethyl starch. Postleukapheresis donor blood lymphocyte counts and serum immunoglobulin levels decreased transiently, but significantly, during each procedure when compared with preleukapheresis values due simply to plasma volume expansion (acute effects of dilution). Possible long‐term effects of repeated procedures were investigated by comparing preleukapheresis values before the first and fourth procedures. Total lymphocyte, total T‐lymphocyte, and T4‐lymphocyte counts, as well as serum immunoglobulin values and anti‐A and B titers measured before the fourth procedure remained unchanged when compared with values obtained before beginning the series of procedures. In contrast, the B‐lymphocyte count was decreased, the T8‐lymphocyte count was increased, and the T4:T8 ratio was decreased due to the increase in the T8‐lymphocyte count. Although statistically significant (p < 0.05), all changes were slight, and clinically significant effects on donor humoral immunity could not be detected. The mechanisms involved, however, may be of biologic and therapeutic interest.
The influence of irradiation on stored platelets
Authors: Moroff G. , George V. M. , Siegl A. M. , Luban N. L. C.
Published: March 2003

Platelet concentrates intended for transfusion to immunosuppressed patients are irradiated to minimize transfusion‐induced graft‐versus‐host disease. Because few reports describe how irradiation influences stored platelets, the authors studied whether 5000 rad of gamma irradiation, the maximum dose currently used clinically, altered platelets in vitro. Platelet concentrates were stored for either 1 day or 5 days in plastic (PL 732) containers before gamma irradiation. One unit of a pair of identical platelet concentrates was irradiated; the second unit served as a control. Irradiation did not alter platelet morphology, mean platelet volume, expression of platelet‐factor‐3 activity, response to hypotonic stress, extent of discharge of lactate dehydrogenase, release of β‐thromboglobulin, formation of thromboxane B2, nor the ability to undergo synergistic aggregation. The lack of any substantial change was observed whether the platelet concentrates were stored initially for either 1 day or 5 days. These results suggest that stored platelets are not altered deleteriously by irradiation with 5000 rad.
Effects of AS‐3 nutrient‐additive solution on 42 and 49 days of storage of red cells
Authors: Simon T. L. , Marcus C. S. , Myhre B. A. , Nelson E. J.
Published: March 2003

Storage of red blood cells in a nutrient‐additive solution, AS‐3 (Nutricel, Cutter Biological, Berkeley, CA), was evaluated after 42 and 49 days of storage by in vitro measurements of hemolysis, adenosine triphosphate (ATP) levels, glucose levels and other constituents, and in vivo study of 24‐hour survival of autologous, reinfused red cells labeled with 51Cr with and without 125I human serum albumin. Two laboratories conducted the studies independently. After 42 days of storage, hemolysis was within an acceptable range (0.72 +/− 0.4% and 1 +/− 0.2%), ATP decreased to 61 percent and 56 percent in the two laboratories, and 24‐hour survivals were 85.1 +/− 8.3 percent for single‐label cells in one laboratory and 82.8 +/− 10 percent (single‐ label cells) and 84.1 +/− 13.1 percent (double‐label cells) in the second laboratory. Thus, results for single‐ and double‐label cells were similar. After 49 days of storage, ATP fell to 45 and 46 percent in the two laboratories. Twenty‐four‐hour recovery fell to 69.4 +/− 7.4 percent with single‐label cells and to 68.2 +/− 6.7 percent with double‐ label cells in one laboratory. In the other laboratory, a paired study comparing AS‐3 with the already approved AS‐1 solution (Adsol; Fenwal Laboratories, Deerfield, IL) showed nearly identical 24‐hour survivals of 71.9 +/− 8.8 percent in Nutricel and 71.8 +/− 6.5 percent in Adsol. These studies demonstrate the excellent viability of the new solution after 42 days of study. At 49 days of study, viability decreased significantly and was comparable in the two nutrient‐additive solutions studied. The value of paired comparison study is demonstrated by the latter results.
Neutralization of P blood group antibodies by synthetic solid‐phase antigens
Authors: Cowles J. W. , Blumberg N.
Published: March 2003

P blood group system antibodies are encountered frequently in clinical transfusion practice. The authors describe an improved method for removing P antibodies from patient specimens without dilution. Solid‐ phase synthetic P antigens were used to neutralize serums containing P blood group system antibodies; 26 alloantibodies with specificity other than P were not inhibited. The synthetic P antigens are also used to characterize the heterogeneous immunochemical fine specificity of antibodies to P1 and P + P1 + Pk.
Evaluation of a centrifugal blood cell processor for washing platelet concentrates
Authors: Vesilind G.W. , Simpson M.B. , Shifman M.A. , Colman R.E. , Kao K.J.
Published: March 2003

A semiautomated saline wash procedure using a blood cell processor was evaluated as a technique for removing plasma from platelet concentrates. In vitro studies demonstrated 92 to 99.6 percent (mean, 96%) removal of total plasma protein (n = 30) with 84 to 97 percent (mean, 90.8%) platelet recovery (n = 28) in post‐wash units. Post‐wash pH values changed by +0.2 to −0.86 (mean, −0.47) (n = 30); the level of recovery from hypotonic shock was 69 to 97 percent (mean, 86%) (n = 11) of pre‐wash units; weighted morphology scores decreased from a mean of 248 to 223 (n = 9). Aggregation response to arachidonic acid, collagen, and adenosine diphosphate plus epinephrine showed essentially no change following the wash procedure, and electron microscopy demonstrated slight morphologic alteration. Autologous platelets labeled with indium‐ 111 demonstrated 43 +/− 20 percent recovery (n = 11) for washed units, compared to 41 +/− 10 percent for control unwashed units (n = 5); mean survivals were 140 +/− 41 hours (n = 11) for washed platelets and 185 +/− 28 hours for unwashed units (n = 5). Thirteen alloimmunized patients receiving 55 washed platelet concentrates demonstrated a mean 1‐ to 4‐hour corrected count increment of 3.99 × 10(3) per microliter, compared to 3.02 × 10(3) per microliter for 77 unwashed platelet units given to the same patients. This study documents that platelet concentrates maintain viability and efficacy following a semiautomated saline wash method using the Cobe 2991 Blood Cell Processor, a technique that may be helpful for patients who require plasma‐depleted platelet transfusions.
Quality assessment of transfusion practice in elective surgery
Authors: Giovanetti A.M. , Parravicini A. , Baroni L. , Riccardi D. , Pizzi M.N , Almini D. , Sirchia G.
Published: March 2003

A program of quality assurance (QA) was adopted to improve blood transfusion practice in elective surgery at a large urban hospital. For this purpose, a cooperative multidisciplinary group was formed, key indicators were identified, and an organization was set up. Data collected by this organization in the 1‐year period needed for implementation of the program indicated that blood misuse was common practice. In fact, overrequest, overtransfusion, excessive reconstitution of whole blood (i.e., concurrent transfusion of red cells and fresh‐frozen plasma), and underuse of predeposit were found in all ten surgical departments of the hospital. In a pilot study, data were collected from one surgical department during and after the implementation phase of the QA program; comparison of these data showed a postimplementation reduction of about two thirds in overtransfusion, whereas overrequest, reconstitution of whole blood, and predeposit rates remained unchanged. These results prompted continuation of the program in order to reach a definitive evaluation of its effectiveness.
Ascorbate‐2‐phosphate in red cell preservation
Authors: Moore G. L. , Marks D. H. , Carmen R. A. , Ledford M. E. , Nelson E. J. , Fishman R. J. , Leng B. S.
Published: March 2003

A red cell additive solution (AS‐005) containing ascorbate‐2‐phosphate (AsP) to maintain 2,3‐diphosphoglycerate, plus adenine, phosphate, and mannitol to retain viability and reduce hemolysis, was evaluated by human clinical trials. A crossover design was used with another additive solution (Nutricel AS‐3, Cutter Laboratories) serving as the control for each donor. Each additive solution was evaluated at 35 and 42 days of storage. There was no significant difference between the red cell viability of the two storage solutions at either time period. Split‐bag, AS‐005 in vitro studies at two temperatures (2.5 and 5.5° C), both within the range of 1 to 6° C approved by the American Association of Blood Banks and the Food and Drug Administration, resulted in dramatically different in vitro parameters, including a threefold difference in 2,3‐diphosphoglycerate (2,3‐DPG), a fivefold difference in glucose, and significant differences in pH and adenosine triphosphate. High‐pressure liquid chromatography data confirmed the preliminary report that 1 to 2 percent (wt/wt) oxalate was present in preparations of AsP. In vitro storage data confirmed that oxalate is the active component of AsP that preserves 2,3‐DPG during storage.
Adherence of fresh and stored granulocytes to endothelial cells. Effect of storage temperature
Authors: Lane T. A. , Lamkin G. E.
Published: March 2003

Granulocyte (PMN) concentrates collected for transfusion to septic, neutropenic patients are stored in the blood bank for various periods of time before they are given. Current methods of blood bank storage of PMN concentrates are associated with impaired in vitro PMN chemotaxis (CTX) and in vivo recovery and circulation kinetics after 24 hours of storage. This suggested the possibility that PMN may become hyperadherent during storage. To test this hypothesis, PMN concentrates were harvested and stored at both 22 and 6° C and their adherence properties to relevant biologic surfaces, endothelial cell (EC) monolayers, and extracellular matrix (ECM) derived from endothelium were measured. Adherence was measured within 4 hours of collection and after 24 and 48 hours of storage. The aggregation properties of fresh and stored PMN were also studied. The adherence of fresh, unstimulated PMN to EC and ECM (31 ± 5% and 34 ± 4%, respectively) increased significantly after storage for 24 hours (EC = 41 ± 8%; ECM = 43 ± 4%) at 22° C. F‐Met‐Leu‐Phe (FMLP) stimulated the adherence of fresh PMN (EC = 37 ± 4%; ECM = 42 ± 4%; p < 0.05). The adherence of PMN stored at 22° C was further stimulated by FMLP (EC = 46 ± 6%; ECM = 50 ± 4%). PMN stored at 6° C had significantly higher adherence than PMN stored at 22° C, and the percentage of increase in adherence induced by FMLP was attenuated in PMN stored at 6° C. Also in contrast to PMN stored at 22° C, PMN stored at 6° C aggregated spontaneously when warmed to 37° C and had an apparent decrease in their aggregation responses to FMLP. These studies indicate that short‐term liquid storage of PMN is accompanied by the development of hyperadherence and a tendency to spontaneous aggregation, particularly in PMN stored at 6° C. These changes may be responsible for the diminished in vitro PMN chemotactic function and the in vitro recovery, circulation, and migration of PMN after storage.
Rh antibodies against the pretransplant red cells following Rh‐incompatible bone marrow transplantation
Authors: Heim M.U. , Schleuning M. , Eckstein R. , Huhn D. , Siegert W. , Clemm C. , Ledderose G. , Kolb H.J. , Wilmanns W. , Mempel W.
Published: March 2003

A 22‐year‐old, blood group O, Rh‐positive (R2r) man received bone marrow from his blood group A, Rh‐negative (rr), HLA‐identical sister for treatment of acute lymphocytic leukemia. The patient's pretransplantation serum contained anti‐A in a low concentration; therefore, plasmapheresis was not done prior to transfusion of bone marrow. To prevent graft‐versus‐host disease, bone marrow was incubated with absorbed rabbit antithymocyte globulin prior to infusion, and the patient was treated with methotrexate in the posttransplantation period. After transplantation, the patient received 6 units of group O, Rh‐negative (rr) packed red cells from random donors and 6 units of platelets from the marrow donor. Three months after transplantation, 0.5 percent of his red cells were still of the host's type (group O, Rh‐positive), as detected by immunofluorescence technique in blood smears. Four months after transplantation, three different Rh antibodies—anti‐D, ‐E, and ‐G—were detected. Since the patient received only Rh‐negative red cell transfusions, it is concluded that he was immunized to his original red cells.
Follow‐up testing and notification of anti‐HIV Western blot atypical (indeterminant) donors
Authors: Kleinman Steven , Fitzpatrick Lynda , Secord Karen , Wilke Diana
Published: March 2003

ABSTRACT: We conducted anti‐HIV testing on follow‐up samples obtained at a mean interval of 20 weeks from 150 blood donors who had previously tested anti‐HIV ELISA positive and Western blot atypical. Of 93 donors who demonstrated reactivity to HIV core protein p24, 4 progressed to positive Western blots. Most of the remaining donors showed a persistent p24 reactivity on Western blot and had no risk factors for HIV infection. Immunofluorescence testing of the initial sample from 93 donors could not definitively separate seroconverters from those with persistent p24 reactivity. Of 57 donors with p18 reactivity, none were positive on follow‐up anti‐HIV testing. Our findings suggest policies and strategies for notifying donors of atypical anti‐HIV Western blot results.
Long‐term frequent plasma exchange donation of cryoprecipitate
Authors: McLeod B.C. , Sassetti R.J. , Cole E.R. , Scott J.P.
Published: March 2003

Plasma exchange donation accomplishes the selective donation of cryoprecipitate. It facilitates the repeated donation of large quantities of factor VIII by individual donors and reduces donor exposure for recipients. A highly motivated donor is described who has undergone 103 donations between May 1983 and March 1987, producing 359,460 IU of factor VIII and supplying all the factor VIII needed since August 1983 by his severely affected hemophiliac son, now age 14. The donor has remained in good health, and no significant abnormalities have been noted in hematologic, biochemical, immunologic, coagulation, and serum protein testing. Extensive experience with this donor suggests that repeated plasma‐exchange donation is safe and can sometimes allow single‐donor support of severe hemophiliacs.
First example of familial posttransfusion purpura in two PlA1‐negative sisters
Authors: Chaplin H. , Aster R.H. , Morgan L.K. , Horn B. , McDonagh K.T. , Baskin R.
Published: March 2003

Posttransfusion purpura (PTP) (platelet count 5000/μl) was diagnosed in a female patient (never transfused, gravida IV, para IV) 1 week after transfusion for hysterectomy in 1978. She did not respond to pooled random‐donor platelets but recovered following a single plasma exchange. Her platelets were PlA1 negative, and her plasma contained potent anti‐PlA1. In 1986, her sister (never transfused, gravida III, para III) developed PTP (platelet counts 5–15,000/μl) following surgery‐associated transfusion. She did not respond to pooled random‐donor platelets. Platelet‐associated IgG was markedly elevated (5365) molecules/platelet; normal, < 660); her plasma contained a potent platelet antibody with anti‐PlA1 specificity. Her platelets were subsequently shown to be PlA1 negative. The platelet count did not rise above 30,000 per μl, despite 3 days of high‐dose methylprednisolone sodium succinate and 2 weeks of prednisone (80 mg/day). Later, her platelet count increased and remained normal after steroids were discontinued. The two sisters proved to be HLA‐identical, and each possessed one haplotype carrying the DR3 marker, which has been implicated as a risk factor in neonatal alloimmune thrombocytopenia associated with anti‐PlA1.
Volume control of erythrocytes during storage
Authors: Beutler E. , Kuhl W.
Published: March 2003

When red cells are stored as suspensions in artificial preservative solutions such as those consisting of saline, adenine, and glucose, increased hemolysis of the red cells occurs in vitro, a phenomenon that is readily prevented by the addition of mannitol. The mechanism by which mannitol prevents hemolysis is unknown. The authors have examined the possibility that mannitol prevents hemolysis by prevention of osmotic swelling of red cells that might otherwise increase their volume beyond their critical hemolytic volume. Storage in artificial media does, indeed, result in swelling of the red cells, which is prevented by mannitol and is increased by the addition of nystatin, an agent that greatly increases the sodium‐potassium permeability of the cells. Swelling and hemolysis are correlated at different concentrations of mannitol, but the correlation breaks down when the case of nystatin‐treated cells is considered; even with marked swelling, there is relatively little hemolysis. Moreover, increasing the volume of the stored red cells acutely by osmotic means results in relatively little additional hemolysis. These findings make it appear unlikely that simple osmotic lysis is the cause of hemolysis in stored cells. Some other stabilizing effect of mannitol that may or may not be related to its osmotic effect must play a role.