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Abstract Background Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processes has been considerably more difficult. Results In this article, we describe a novel technology for generating cell membrane microarrays for performing high throughput biology. Our robotically-arrayed supported membranes are physiologically fluid, a critical property which differentiates this technology from other previous membrane systems and makes it useful for studying cellular processes on an industrialized scale. Membrane array elements consist of a solid substrate, above which resides a fluid supported lipid bilayer containing biologically-active molecules of interest. Incorporation of transmembrane proteins into the arrayed membranes enables the study of ligand/receptor binding, as well as interactions with live intact cells. The fluidity of these molecules in the planar lipid bilayer facilitates dimerization and other higher order interactions necessary for biological signaling events. In order to demonstrate the utility of our fluid membrane array technology to ligand/receptor studies, we investigated the multivalent binding of the cholera toxin B-subunit (CTB) to the membrane ganglioside GM1. We have also displayed a number of bona fide drug targets, including bacterial endotoxin (also referred to as lipopolysaccharide (LPS)) and membrane proteins important in T cell activation. Conclusion We have demonstrated the applicability of our fluid cell membrane array technology to both academic research applications and industrial drug discovery. Our technology facilitates the study of ligand/receptor interactions and cell-cell signaling, providing rich qualitative and quantitative information.

Background and Objectives: The World Health Organization (WHO) African Region accounts for 94% of malaria cases globally, with variability recognized within endemic regions. To determine the detection rate of Plasmodium RNA in blood donors resident in malaria-endemic areas, samples from three African countries were tested using a Plasmodium nucleic acid test., Materials and Methods: Whole blood (WB) samples collected from routine donors in Cameroon, Madagascar and Mali were shipped frozen to the United States. Samples were tested individually from WB lysates with the Procleix Plasmodium assay (transcription-mediated amplification [TMA]). Reactive samples were considered either repeat reactive or initial reactive only, depending on TMA-retest results. When available, matching plasma samples were tested for Plasmodium antibodies by enzyme immunoassay (EIA)., Results: Plasmodium repeat reactivity ranged from 41% (91/223 tested) in Cameroon to 12% (26/216) in Mali and 1% (3/249) in Madagascar. Initially reactive samples, where reactivity did not repeat, were identified from Cameroon (5/223; 2%) and Mali (2/216; 1%). The matched-plasma subgroup had EIA reactivity ranging from 86% (113/131 tested) in Cameroon to 59% (10/17) in Mali and 27% (68/248) in Madagascar., Conclusion: Plasmodium ribosomal RNA (rRNA) detection and antibody rates varied greatly in the three countries studied. Detection of Plasmodium rRNA can provide an additional tool to address malaria risk in blood donors., (© 2024 International Society of Blood Transfusion.)

Background: The first coronavirus disease 2019 (COVID-19) case in the United States was recognized on 19 January 2020, but the time of introduction of the virus into the United States is unknown. An existing sample cohort was examined for serologic evidence of early severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections., Methods: A repository of 46 120 samples from healthy routine blood donors, representing 46 states and the District of Columbia, was tested for total antibodies to SARS-CoV-2 nucleocapsid (anti-N) using a commercial test. All reactive samples were further tested using an experimental receptor-binding domain (RBD)-specific immunoglobulin G (IgG) enzyme-linked immunosorbent assay. Further testing was also conducted for anti-spike (anti-S) antibodies by commercial tests, experimental anti-S immunologic blocking, and for antibodies to the 4 human cold coronaviruses., Results: Anti-N reactivity was observed in 92 tested samples (0.2%), 91 of which had adequate volume for further testing; of these, 55 were confirmed positive by anti-RBD. None of these reactive findings were attributable to the other human coronaviruses tested. The confirmed-positive frequency increased over time paralleling patterns observed for COVID-19 cases reported in the United States (in contrast to stable patterns over time for the cold coronaviruses). Nine confirmed positive samples (0.07%) were identified among the 13 364 donations collected between 13 December 2019 and 22 January 2020. None of these early confirmed-positive samples were reactive by commercial anti-S tests suggesting very recent infection., Conclusions: The samples tested in this study were broadly representative of the United States, and all were from individuals who had successfully donated blood. The antibody-reactive results of this study suggest that SARS-CoV-2 was likely present in the United States before 19 January 2020., (© The Author(s) 2024. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Background: Malaria is caused by protozoa of the genus Plasmodium and transmitted by Anopheles mosquitos. In the US, blood donors are assessed for malaria risk, including donor travel or previous residence in endemic areas and history of malaria by questionnaire and deferred for three months or three years, respectively., Methods: The Procleix Plasmodium Assay is a qualitative nucleic acid test based on transcription-mediated amplification (TMA) for the detection of 18S ribosomal RNA of P. falciparum, P. ovale, P. vivax, P. malariae, and P. knowlesi for use on the Procleix Panther system. Analytical sensitivity was evaluated with in vitro transcripts and infected red blood cells. For clinical specificity, 12,800 individual donations and 283 pools of 16 samples from routine US donors were screened. Malaria risk was evaluated by testing 862 donors deferred for 3 years. Reactive results were confirmed with in-house real-time TMA assay and serology., Results: Assay sensitivity was 8.47-11.89 RNA copies/mL and 2.10-6.82 infected red cells/mL. Specificity was 99.99% in 12,800 individual donations and 100% in 283 pools of 16. Of 862 tested deferred donor samples, one donor (0.12%) confirmed positive individually and in pools; he remained confirmed positive for 13 months. The infected donor was a prior resident of a malaria-endemic area in West Africa., Conclusions: The Procleix Plasmodium Assay showed high sensitivity and specificity and detected Plasmodium RNA in an asymptomatic presenting donor. This assay may prove helpful as a screening test versus the use of risk questions to reduce the number of donors deferred for malaria risk., (© 2023 AABB.)

Background: The 2022 multi-country outbreak of monkeypox (mpox) resulted in blood collection and public health agencies closely monitoring for changes in transmission dynamics that could pose a threat to the blood supply. While mpox virus (MPXV) is not known to be transfusion transmissible, there have been several studies demonstrating the detection of MPXV in blood. We evaluated the performance characteristics of a research use only (RUO) nucleic acid amplification test for MPXV. The assay was developed to detect MPXV DNA in plasma and serum specimens from human blood donors., Methods and Materials: The sensitivity of the RUO MPXV Assay was determined using a synthetic DNA sequence, purified full-length genomic DNA, and a chemically inactivated virus. Specificity was determined using fresh plasma samples collected from blood donors during the outbreak. Plasma samples collected from donors considered at increased risk for exposure to mpox were also tested., Results: For sensitivity, the 95% limit of detection (LOD) ranged from 0.26 copies/mL (inactivated virus) to 31.65 copies/mL (synthetic DNA) to 166.61 copies/mL (for full-length DNA). All donor samples tested with the RUO MPXV Assay were nonreactive, resulting in a specificity of 100% (95% CI, 99.93%-100.00%)., Discussion: The RUO MPXV Assay was developed as a potential blood donation screening assay in response to the outbreak. While not directly comparable, the 95% LOD fiducial limits obtained from partial- and full-length DNA analysis were similar to other manufacturers' MPXV assays. Additionally, this assay demonstrated high specificity for screening blood donors., (© 2023 AABB.)

Background: This study evaluated whether pathogen reduction technology (PRT) in plasma and platelets using amotosalen/ultraviolet A light (A/UVA) or in red blood cells using amustaline/glutathione (S-303/GSH) may be used as the sole mitigation strategy preventing transfusion-transmitted West Nile (WNV), dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viral, and Babesia microti, Trypanosoma cruzi, and Plasmodium parasitic infections., Methods: Antibody (Ab) status and pathogen loads (copies/mL) were obtained for donations from US blood donors testing nucleic acid (NAT)-positive for WNV, DENV, ZIKV, CHIKV, and B. microti. Infectivity titers derived from pathogen loads were compared to published PRT log 10 reduction factors (LRF); LRFs were also reviewed for Plasmodium and T. cruzi. The potential positive impact on donor retention following removal of deferrals from required questioning and testing for WNV, Babesia, Plasmodium, and T. cruzi was estimated for American Red Cross (ARC) donors., Results: A/UVA and S-303/GSH reduced infectivity to levels in accordance with those recognized by FDA as suitable to replace testing for all agents evaluated. If PRT replaced deferrals resulting from health history questions and/or NAT for WNV, Babesia, Plasmodium, and T. cruzi, 27,758 ARC donors could be retained allowing approximately 50,000 additional donations/year based on 1.79 donations/donor for calendar year 2019 (extrapolated to an estimated 125,000 additional donations nationally)., Conclusion: Pathogen loads in donations from US blood donors demonstrated that robust PRT may provide an opportunity to replace deferrals associated with donor questioning and NAT for vector-borne agents allowing for significant donor retention and likely increased blood availability., (© 2022 The Authors. Transfusion published by Wiley Periodicals LLC on behalf of AABB.)

Background: In 2019, the Food and Drug Administration revised the requirement for further testing of anti-HCV-reactive donations testing nucleic acid (NAT)-nonreactive via routine mini-pool (MP)-NAT. Individual donation (ID)-HCV NAT was required as a supplemental test prior to a second FDA-licensed anti-HCV assay; if ID-HCV-NAT is reactive, no further testing is required. This study investigated the rate of low-level RNA in anti-HCV-reactive donation samples prior to and following the implementation of supplemental ID-HCV NAT., Study Design/methods: A retrospective analysis was conducted on frozen plasma unit samples from 1161 anti-HCV confirmed-positive/HCV-NAT-nonreactive donations collected from December 2014 to January 2020. Samples were tested by multiple replicates on the Grifols Procleix Ultrio Elite (UE) assay and corresponding discriminatory HCV (dHCV) assay on the Procleix Panther System. Prospectively, the prevalence of low-level RNA in 2912 anti-HCV-reactive donations detected through routine screening from April 2020 through March 2021 was determined., Results: In retrospective analyses, 10 (0.86%) of 1161 plasma samples were UE reactive, of which four (0.34%) were dHCV reactive (in all replicates of UE and dHCV). Of 2912 anti-HCV-reactive donation samples testing NAT-nonreactive via routine MP-NAT, three (0.1%; 95% CI: 0.04-0.30) were dHCV reactive; 37% of the remaining samples were reactive by a second anti-HCV assay and thus serologically confirmed., Conclusions: Retrospective and prospective analysis in comparison to earlier studies revealed that low-level HCV-RNA reactivity is decreasing over time. The very low HCV-RNA rates may be due to the widespread use of highly effective, direct-acting anti-viral treatments for those diagnosed with HCV infection., (© 2021 AABB.)

Background: SARS-CoV-2 RNA prevalence in blood donors from large geographic areas of high community transmission is limited. We tested residual donor plasma minipools (MPs) to determine SARS-CoV-2 RNAemia prevalence in six United States areas., Study Design/methods: Blood donations collected from 7 March 2020 to 25 September 2020 were tested for SARS-CoV-2 RNA (vRNA) in MP of 6 or 16 donations using the Grifols Procleix SARS-CoV-2 research-use only (RUO) transcription-mediated amplification (TMA) assay. Reactive results were confirmed using an alternate target region TMA assay. Reactive MPs were tested by TMA after serial dilution to estimate viral load. Testing for anti-SARS-CoV-2 antibodies and infectivity was performed., Results: A total of 17,995 MPs corresponding to approximately 258,000 donations were tested for vRNA. Three confirmed reactive MP16 were identified. The estimated prevalence of vRNA reactive donations was 1.16/100,000 (95% CI 0.40, 3.42). The vRNA-reactive samples were non-reactive for antibody, and the estimated viral loads of the (presumed single) positive donations within each MP ranged from <1000 to <4000 copies/ml. When tested, no infectivity was observed in inoculated permissive cell cultures., Discussion: Blood donation MP-nucleic acid testing (NAT) indicated that SARS-CoV-2 RNAemia is infrequent and, when detected, the vRNA was at low concentrations. Only one RNA-reactive MP could be tested for infectivity for operational reasons and was not infectious in cell culture. These findings support current recommendations from international and national regulatory agencies to not screen donors by NAT., (© 2021 AABB.)

Background: Rare transfusion-transmitted West Nile virus (WNV) cases usually occur due to gaps in testing involving converting to more sensitive nucleic acid testing (NAT) formats (referred to as triggering). Using data from 2014 to 2018, we investigated a strategy used to increase detection early in the triggering period and reviewed its yield as the individual donation (ID)-NAT geographic area was decreased., Methods: Mini-pool-NAT transitioned to ID-NAT following triggering based on one WNV NAT-reactive donation (having an elevated signal, repeat reactive, or in an area with WNV ongoing activity). ID-NAT-triggered geographic areas included an entire state (2014-2017) or collections within a 50-mile radius of the triggering donor's residential zip code (2018). During the MP- to ID-NAT transition, donation samples were retrieved and tested by ID-NAT for those with results not yet released (referred to as in-process testing). Reactive sample confirmation was performed by repeat NAT of an independent sample or antibody testing., Results: ID-NAT included 3.2 million donations of more than 25 million tested year-round, resulting in 684 confirmed positives; all confirmed-positive donations occurred from June to December (0.64/10,000). Overall, 52% (358/684) required ID-NAT for detection, including 68 (19%) antibody negatives. Ten of 19 (53%) identified in-process were ID-NAT-only detectable, including four antibody negatives, or approximately 1 per year (2.8% of ID-NAT-only detectable). With reduced triggering geography, 12 of 19 (63%) were not identified (including 6/10 ID-NAT-only detectable, and 2/4 ID-NAT-only detectable/antibody negative)., Conclusion: WNV NAT's utility is between June-December; however, abandoning testing outside of this time may increase risk. While in-process testing identified approximately one ID-NAT-only detectable (antibody-negative) donation per year, reducing the geographic triggered area decreased its effectiveness., (© 2019 AABB.)

Background: US blood donors are tested for Trypanosoma cruzi antibodies only at their first presentation, based on studies, reviewed here, demonstrating the absence of incident infections. Reports of autochthonous human transmissions of the parasite in Texas have raised concern about the safety of one-time testing., Methods: Positive donation frequencies were evaluated among first-time blood donations from 2007 to 2015. Rates and their temporal changes were evaluated in an area of high T. cruzi infection and compared with rates elsewhere. Donors with positive results were surveyed for risk factors and relevant demographic characteristics., Results: Data from 9.1 million first-time donations were analyzed; 585 (1:15,544) were confirmed positive by radioimmunoprecipitation assay (RIPA) or concordantly positive with a second screening test/licensed assay. Seroprevalence in first-time donors in Southern California (an area of high endemicity) was 1:2,747, or 5.7-fold higher than the overall rate. Rates did not change over time nationally but showed a nonsignificant consistent downward trend in Southern California. The majority (92%) of donors who responded to a questionnaire had one or more T. cruzi endemic-area risk factors. Five donors with likely autochthonous infection were identified (2007-2013); nine additional donors had RIPA false positivity., Conclusion: T. cruzi seroprevalence among donors nationally and in an area of high enzootic infection were stable or declining. Almost all interviewed seropositive donors had known risk factors indicating likely infection years earlier while residing in T. cruzi-endemic areas. In the United States, there was no evidence of increased T. cruzi prevalence among first-time donors., (© 2018 AABB.)

Background: Characteristics of US blood donors with recent (RBI) or occult (OBI) hepatitis B virus (HBV) infection are not well defined., Methods: Donors with RBI and OBI were identified by nucleic acid and serologic testing among 34.4 million donations during 2009-2015. Consenting donors were interviewed and their HBV S-gene sequenced., Results: The overall rate of HBV-infected donors was 7.95 per 100,000; of these, 0.35 per 100,000 and 1.70 per 100,000 were RBI and OBI, respectively. RBI (n = 120) and OBI (n = 583) donors constituted 26% of all HBV-infected (n = 2735) donors. Detection of HBV DNA in 92% of OBI donors required individual donation nucleic acid testing. Donors with OBI compared to RBI were older (mean age, 48 vs 39 years; p < 0.0001) with lower median viral loads (9 vs. 529 IU/mL; p < 0.0001). A higher proportion of OBI than RBI donors were born or resided in an endemic country (39% vs. 5%; p = 0.0078). Seventy-seven percent of all RBI and OBI donors had multiple sex partners, an HBV-risk factor. Of 40 RBI and 10 OBI donors whose S gene was sequenced, 33 (83%) and 6 (60%), respectively, carried HBV subgenotype A2; 18 (55%) and 2 (33%), respectively, shared an identical sequence. Infection with 1 or more putative HBV-immune-escape mutants was identified in 5 (50%) of OBI but no RBI donors., Conclusion: RBI and OBI continue to be identified at low rates, confirming the importance of comprehensive HBV DNA screening of US blood donations. HBV-infected donors require referral for care and evaluation and contact tracing; their HBV strains may provide important information on emergent genotypes., (© 2018 AABB.)

The mammalian homolog of Drosophila empty spiracles 2 (Emx2) is a homeobox transcription factor that plays central roles in early development of the inner ear, pelvic and shoulder girdles, cerebral cortex, and urogenital organs. The role for Emx2 is best understood within the context of the development of the neocortical region of the cortex, where Emx2 is expressed in a high posterior-medial to low anterior-lateral gradient that regulates the partitioning of the neocortex into different functional fields that perform discrete computational tasks. Despite several lines of evidence demonstrating an Emx2 concentration-dependent mechanism for establishing functional areas within the developing neocortex, little is known about how Emx2 physically carries out this role. Although several binding partners for Emx2 have been identified (including Sp8, eIF4E, and Pbx1), no screens have been used to identify potential protein binding partners for this protein. We utilized a yeast two-hybrid screen using a library constructed from embryonic mouse cDNA in an attempt to identify novel binding partners for Emx2. This initial screen isolated two potential Emx2-binding partner proteins, Cnot6l and QkI-7. These novel Emx2-binding proteins are involved in multiple levels of mRNA metabolism that including splicing, mRNA export, translation, and destruction, thus making them interesting targets for further study.

O-linked β-N-acetylglucosamine (O-GlcNAc) is an essential regulatory post-translational modification of thousands of nuclear, cytoplasmic, and mitochondrial proteins. O-GlcNAc is dynamically added and removed from proteins by the O-GlcNAc transferase and the O-GlcNAcase (OGA), respectively. Dysregulation of O-GlcNAc-cycling is implicated in the etiology of numerous diseases including tumorigenesis, metabolic dysfunction, and neurodegeneration. To facilitate studies focused on the role of O-GlcNAc and OGA in disease, we sought to identify commercially available antibodies that enable the enrichment of full-length OGA from lysates of mouse and human origin. Here, we report that antibodies from Abcam and Bethyl Laboratories can be used to immunoprecipitate OGA to near-saturation from human and mouse cell lysates. However, Western blotting analysis indicates that both antibodies, as well as three non-commercially available antibodies (345, 346, 352), detect full-length OGA and numerous cross-reacting proteins. These non-specific signals migrate similarly to full-length OGA and are detected robustly, suggesting that the use of appropriate controls is essential to avoid the misidentification of OGA., (© The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The dynamic post-translational modification O- linked β- N -acetylglucosamine ( O -GlcNAc) regulates thousands of nuclear, cytoplasmic, and mitochondrial proteins. Cellular stress, including oxidative stress, results in increased O- GlcNAcylation of numerous proteins, and this increase is thought to promote cell survival. The mechanisms by which the O- GlcNAc transferase (OGT) and the O- GlcNAcase (OGA), the enzymes that add and remove O- GlcNAc, respectively, are regulated during oxidative stress to alter O- GlcNAcylation are not fully characterized. Here, we demonstrate that oxidative stress leads to elevated O- GlcNAc levels in U2OS cells but has little impact on the activity of OGT. In contrast, the expression and activity of OGA are enhanced. We hypothesized that this seeming paradox could be explained by proteins that bind to and control the local activity or substrate targeting of OGA, thereby resulting in the observed stress-induced elevations of O- GlcNAc. To identify potential protein partners, we utilized BioID proximity biotinylation in combination with s table i sotopic l abeling of a mino acids in c ell culture (SILAC). This analysis revealed 90 OGA-interacting partners, many of which exhibited increased binding to OGA upon stress. The associations of OGA with fatty acid synthase (FAS), filamin-A, heat shock cognate 70-kDa protein, and OGT were confirmed by co-immunoprecipitation. The pool of OGA bound to FAS demonstrated a substantial (∼85%) reduction in specific activity, suggesting that FAS inhibits OGA. Consistent with this observation, FAS overexpression augmented stress-induced O- GlcNAcylation. Although the mechanism by which FAS sequesters OGA remains unknown, these data suggest that FAS fine-tunes the cell's response to stress and injury by remodeling cellular O- GlcNAcylation., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)

Background: Transfusion-transmitted West Nile virus (WNV) infection is infrequent as a result of minipool (MP) and individual-donation (ID) nucleic acid testing (NAT) of blood donations. ID-NAT is triggered on the basis of local WNV activity identified by MP-NAT., Study Design and Methods: A 78-year-old male patient who was treated for cardiac disease received 14 blood components from 30 donors in August 2016. He was discharged 7 days after aortic valve replacement and coronary bypass surgery, but was re-admitted on Day 12 with symptoms of viral infection, and eventually was diagnosed with aseptic meningitis. The patent died on Day 51., Results: The patient was positive for WNV-immunoglobulin M (IgM) antibodies in his cerebrospinal fluid on Day 14 and was positive for WNV-IgM (on Days 14 and 16) and WNV-IgG antibodies (on Day 16) in his serum. All associated donations were nonreactive by MP-NAT or ID-NAT. However, one MP-NAT was noted to have an elevated (but negative) signal-to-cutoff ratio, and one donor from that MP was subsequently found positive for WNV-IgM and IgG antibodies; the donor was diagnosed with a WNV-like viral syndrome that had an onset 3 to 5 days postdonation. The donor's plasma was transfused 12 days before the patient's onset of WNV-meningoencephalitis. Conversion to ID-NAT was triggered for the region 7 days after the implicated donation, which was associated with the region's first human WNV case., Conclusion: Despite the possibility of mosquito-borne transmission, this was considered to be a case of transfusion-transmitted WNV infection from an MP-NAT-nonreactive donation collected just before triggering conversion to ID-NAT; a rare event recognized once in 84 million donations., (© 2017 AABB.)

O-linked N-acetyl-β-D-glucosamine (O-GlcNAc) is a ubiquitous and dynamic post-translational modification known to modify over 3,000 nuclear, cytoplasmic, and mitochondrial eukaryotic proteins. Addition of O-GlcNAc to proteins is catalyzed by the O-GlcNAc transferase and is removed by a neutral-N-acetyl-β-glucosaminidase (O-GlcNAcase). O-GlcNAc is thought to regulate proteins in a manner analogous to protein phosphorylation, and the cycling of this carbohydrate modification regulates many cellular functions such as the cellular stress response. Diverse forms of cellular stress and tissue injury result in enhanced O-GlcNAc modification, or O-GlcNAcylation, of numerous intracellular proteins. Stress-induced O-GlcNAcylation appears to promote cell/tissue survival by regulating a multitude of biological processes including: the phosphoinositide 3-kinase/Akt pathway, heat shock protein expression, calcium homeostasis, levels of reactive oxygen species, ER stress, protein stability, mitochondrial dynamics, and inflammation. Here, we will discuss the regulation of these processes by O-GlcNAc and the impact of such regulation on survival in models of ischemia reperfusion injury and trauma hemorrhage. We will also discuss the misregulation of O-GlcNAc in diseases commonly associated with the stress response, namely Alzheimer's and Parkinson's diseases. Finally, we will highlight recent advancements in the tools and technologies used to study the O-GlcNAc modification.

Crystallisation of yttrium and other trivalent metal cations of similar radius with racemic N,N'-2-methylpiperazinebis(methylene phosphonic acid) gives a small pore bisphosphonate metal-organic framework solid displaying coordinative and hydrogen bonding and permanent porosity.

We developed the German Adaptation of the Personality Assessment Inventory (PAI; Morey, 1991) under careful consideration of current adaptation literature and guidelines. The adaptation process included the translation of the 344 items into German, a back translation into English as well as the testing of the language equivalence using a bilingual sample. We then standardized the final German version of the PAI for the German population. We compared the American and German norm and reliability data. The observed differences in PAI scale means did not exceed 5 T scores. Internal consistency reliability showed a similar pattern in both language versions, although the German alpha coefficients were on average slightly lower than the American ones. Factor structure was similar in both versions. We discuss expectations about the German PAI and possible problems for its practical usefulness for the German-speaking population.

The first large pore (free diameter > 7 A) metal phosphonates have been prepared as divalent metal N,N'-piperazinebis(methylenephosphonate)s that possess pores greater than ca. 10 A in free diameter, are stable up to 400 degrees C and offer a route to chiral adsorbents and catalysts.

Fibroblast growth factors (Fgfs) have long been implicated in regulating vertebrate skeletal muscle differentiation, but their precise role(s) in vivo remain unclear. Here, we show that Fgf8 signalling in the somite is required for myod expression and terminal differentiation of a subset of fast muscle cells in the zebrafish lateral somite. In the absence of Fgf8, lateral somite cells transiently express myf5 but fail to make muscle and remain in a dermomyotome-like state characterised by pax3 and meox expression. Slow muscle fibres form and commence normal migration in the absence of Fgf8, but fail to traverse the expanded undifferentiated lateral somite. The Fgf8-independent residual population of medial fast muscle fibres is not Hedgehog dependent. However, Fgf8-independent medial fast muscle precursors are lacking in floatinghead mutants, suggesting that they require another ventral midline-derived signal. We conclude that Fgf8 drives terminal differentiation of a specific population of lateral muscle precursor cells within the early somite.

An exploration of the reactions of N,N'-piperazinebis(methylenephosphonic acid), H4L, with zinc salts has led to the isolation of two new framework zinc phosphonates. ZnLH2.H2O (I) is isostructural with previously reported manganese(II) and cobalt(II) analogues, and consists of infinite 'zinc phosphate' chains bridged into three dimensions via the organic moieties. The resulting framework encloses large channels in which the loosely bound H2O resides. The H2O is lost reversibly at around 160 degree C, without framework collapse. Zn2L (II) has a novel framework structure, prepared at an initial pH > 7, which consists of two-dimensional 'zinc phosphate' sheets, comprising both four- and eight-membered -Zn-O-P- rings, which are also linked into three dimensions via the organic groups. In both cases, the zinc centre is tetrahedral; in I coordination is by oxygen atoms from four different phosphonate groups, whereas in II the additional deprotonation of the ligand allows coordination via three oxygen atoms plus the amine nitrogen atom.

Two novel three-dimensional lanthanum coordination polymers have been prepared with the phosphonic acid H2O3PCH2N(C2H4)2 NCH2PO3H2 (LH4). La2(LH2)2(LH4)Cl2 (I) and La2(LH2)2(LH3)Cl (II) arise from similar hydrothermal reactions but differing sources of La. A one-dimensional "lanthanum-phosphate" chain, comprising corner-linked LaO7 and PO3C polyhedra, forms the basis for the two different structures. The two structures differ in the mode of connectivity of the inorganic chains via the phosphonate groups. Both materials include extraframework chloride ions, the different amounts of which apparently direct the polytypic structures. In II, the chloride ions are incorporated in a noncentrosymmetric manner leading to a polar framework topology.

High cell density cultures of CHO cells growing in a bioreactor under dissolved oxygen control were found to undergo spontaneous bifurcations and a subsequent loss of stability some time into the fermentation. This loss of stability was manifested by sustained and amplified oscillations in the bioreactor dissolved oxygen concentration and in the oxygen gas flow rate to the reactor. To identify potential biological and operational causes for the phenomenon, linear stability analysis was applied in a neighborhood of the experimentally observed bifurcation point. The analysis revealed that two steady state process gains, K(P1) and K(P2), regulated k(l)a and gas phase oxygen concentration inputs, respectively, and the magnitude of K(P1) was found to determine system stability about the bifurcation point. The magnitude of K(P1), and hence the corresponding open-loop steady state gain K(OL1), scaled linearly with the bioreactor cell density, increasing with increasing cell density. These results allowed the generation of a fermentation stability diagram, which partitioned K(C)-N operating space into stable and unstable regions separated by the loci of predicted critically stable controller constants, K(C,critical), as a function of bioreactor cell density. This consistency of this operating diagram with experimentally observed changes in system stability was demonstrated. We conclude that time-dependent increases in cell density are the cause of the observed instabilities and that cell density is the critical bifurcation parameter. The results of this study should be readily applicable to the design of a more robust controller., (Copyright 2003 Wiley Periodicals, Inc.)

This paper summarizes the analytical and occupational hygiene findings from a recent survey of occupational exposure to metalworking fluids (MWFs) in the engineering industry. The aim of the survey was to link MWF mist exposure measurements with particular engineering processes and controls, and utilize the data obtained to develop exposure standards. At the same time the opportunity was taken to assess fluid management and control, including bacterial and fines contamination in the machine sumps. In general, occupational exposure to mineral oil MWF mist was controlled to <3 mg/m(3) (8 h time-weighted average) and to <1 mg/m(3) for water-mix MWF mist (in terms of the concentrate). These exposure values do not necessarily represent best practice, but are believed to be achievable and representative of industry as a whole. Gravimetric analysis of the total inhalable particulate was found to be a good predictor of mineral oil MWF mist but not for water-mix MWF mist. Grinding and drilling operations produced higher exposures than turning and milling for water-mix fluids. There were insufficient data to compare machining operations for mineral oil MWFs. On the whole, fluid management was found to be poor, with most sites failing to meet industry good practice or Health & Safety Executive (HSE) standards. Some of the operating procedures utilized were deficient or unsatisfactory. Poor standards of fluid management were found at all sizes of company. High levels of bacteria, endotoxin and fines were found in sumps, and control of other factors, such as water-mix fluid concentration, was often poor. Mineral oils had higher levels of fines than water-mix fluids (medians of 395 and 18 mg/l, respectively), and grinding produced high levels of fines in both types of MWF. Many water-mix sumps contained bacterial levels of >1 x 10(6) CFU/ml, and endotoxin levels of >100 000 EU/ml were not uncommon. The median values were 109 000 CFU/ml and 8039 EU/ml, respectively. Mists could potentially contain extensive contamination from bacteria and endotoxin. Analysis of the data suggests that sumps operating under typical conditions for machining (a temperature of 20 degrees C, a pH of 9 and a fluid strength below 10%), also appear to provide optimum conditions for the proliferation of bacteria. Low levels of benzo[a]pyrene (median 0.03 micro g/g) were found in the mineral oils, and low levels of N-nitrosodiethanolamine (median 0.4 micro g/ml) were found in the water-mix MWFs. The results of this work will contribute to guidance from the HSE, setting out accepted industry good practice, including guide values for MWF mist and sump fluid contaminants, with significant emphasis on sump fluid management (maintenance and monitoring), as well as control issues.

Not all mineral oil metalworking fluids (MWFs) in common use form stable airborne mists which can be sampled quantitatively onto a filter. This much has been known for some time but no simple method of identifying oils too volatile for customary filter sampling has been developed. Past work was reviewed and experiments were done to select simple criteria which would enable such oils to be identified. The sampling efficiency for a range of commercial mineral oil MWF were assessed by drawing clean air through spiked filters at 2 l. min(-1) for periods up to 6 h before analysis. The physical properties of MWF are governed by their composition and kinematic viscosity was found to be the most practical and easily available index of the potential for sample loss from the filter. Oils with viscosities greater that 18 cSt (at 40 degrees C) lost less than 5% of their weight, whereas those with viscosities less than 18 cSt gave losses up to 71%. The losses from the MWF were mostly aliphatic hydrocarbons (C(10)-C(18)), but additives such as alkyl benzenes, esters, phenols and terpene odorants were also lost. The main recommendation to arise from the work is that filter sampling can be performed on mineral oils with viscosities of 18 cSt (at 40 degrees C) or more with little evaporative losses from the filter. However, sampling oils with viscosities less than 18 cSt will produce results which may significantly underestimate the true value. Over a quarter of UK mineral oil MWFs are formulated from mineral oils with viscosities less than 18 cSt (at 40 degrees C). The problem of exposure under-estimation and inappropriate exposure sampling could be widespread. Further work is being done on measurement of mixed phase mineral oil mist exposure.

Background: Alcohol consumption previously has been demonstrated to reduce the density and strength of cortical bone of young, actively growing rats. Osteoblast activity and trabecular bone volume were also significantly lower. A germane question arising from these studies is whether the detrimental effects would persist into adulthood. To address this issue, a long-term study was undertaken with animals that consumed alcohol throughout their life and into old age., Methods: One-month-old female Sprague-Dawley rats were divided into three diet groups: alcohol-fed, pair-fed, and chow-fed. The alcohol-fed animals received a modified Lieber-DeCarli diet that contained 35% ethanol-derived calories. The pair-fed group served as a caloric-equivalent control, and the chow-fed animals served as a completely untreated control. Animals were euthanized after five time periods on the diets that represented three stages of the life span: young (3 months), adult (6, 9, 12 months), and aged (18 months). The left femur was isolated and mechanically tested in 3-point bending for mechanical properties., Results: In the young animals, alcohol consumption produced dramatic reductions in both extrinsic (whole bone) and intrinsic (tissue material) properties, which is consistent with results from previous studies on growing rats. For the adult animals, however, the alcohol groups were only slightly lower and the differences were not statistically significant. The aged animals showed diminished properties due to alcohol, but only for the intrinsic material properties. The extrinsic properties remained similar to controls as a result of greater radial expansion in the femur diaphysis. Despite the cross-sectional areas being the same, this expansion gave rise to higher cross-sectional moment of inertia values in the alcohol animals. The thickness of the cortical wall was lowest in the alcohol group at all time points., Conclusions: Long-term alcohol consumption produced two major effects in the oldest animals studied: the quality of the cortical bone tissue was diminished, as evidenced by reduced elastic modulus and ultimate strength values, and the bone seemed to compensate for this by expanding the cross-section to produce larger cross-sectional moment of inertia values. The reduced bone tissue quality is consistent with the lower ash percent values in the alcohol animals, but other factors such as the quality of the collagen and mineral crystal may also be important contributors.

A method for measuring hydroquinone in air was evaluated, both in the laboratory and in the workplace. The method involved sampling the inhalable fraction onto a filter contained in a multi-holed sampler with a back-up of Tenax TA, followed by desorption into acetonitrile and analysis by High Performance Liquid Chromatography. It was shown to be effective at measuring hydroquinone over the range 0.1 to 2 times a concentration of 0.5 mg/m3 for 8 h.

The behaviour of common alkyl-2-cyanoacrylates in air has been investigated. Increasing humidity was found to increase the rate of polymerisation, although polymerisation is not instantaneous, even at high humidities. Sampling methods for these compounds have been studied and we are able to recommend a pumped Tenax sampling tube, although certain sampling limitations apply, notably that specified flow rates must be applied and that a maximum sampling time of four hours is recommended. Significant errors can occur if sampling is performed outside the range of conditions specified. Upon completion of sampling the tube is desorbed into 0.2% phosphoric acid in acetonitrile. The resulting solution is analysed by HPLC.

Test samples were collected at a range of sites using the MDHS 68 sampler, a 7-hole sampler, and a cyclone respirable-dust sampler. Glass fibre, Zefluor, Fluoropore and silver (0.8 and 5 microns) filters were compared for weighing error, handling characteristics, and amount collected. The effects of a carbon back-up, of extraction solvent, and of analytical method were studied. The current HSE method for the sampling and analysis of coal tar pitch volatiles, MDHS 68 has been reviewed. It is recommended that the MDHS 68 sampler be replaced by a 7-hole head containing a silver or PTFE filter (approximately 1 micron pore size), with a graphitized carbon back-up if efficient collection of 2 to 4-ring PAHs is required. Cyclohexane is recommended as the extraction solvent. Methods of analysis which quantify the fume as a whole (MDHS 68, u.v., ELSD) were shown to give a response which varied according to the type of CTPV generating process. Therefore analysis of specific PAHs of occupational interest, by GC-MS or HPLC is recommended.

Object: Diesel soot has been recognized as probably carcinogenic to humans. Elemental carbon (also called black carbon) in soot is considered at the moment as the most significant surrogate to be measured for assessing the exposure to this pollutant. Its analysis is done by combustion in an oven and determination of the CO2 formed, after elimination of the organic fraction of the soot by heating and/or by solvent extraction. The analysis allows determination of both fractions of the soot: "elemental carbon" (EC) and organic carbon (OC). The sum of EC and OC is called TC (total carbon)., Method: An informal European coordination group organized two round robin tests on filter samples collected from diluted diesel emissions. The first round (RRT1) was performed on 13 different samples analyzed by ten laboratories. The range of loading was 2.5 to 150 micrograms/cm2 of EC. No evaluation of the precision within laboratories could be made since each laboratory gave only one result per sample. Therefore a second round (RRT2) was organized with two samples and a blank filter sent in several portions to 11 laboratories. It should be stressed that each laboratory used its own method and that no standardization was planned at this stage., Results: Results of RRT1 showed that the coefficient of variation between laboratories decreased with higher loading and was around 10% to 15% for EC above about 20 micrograms/cm2. Dispersion of the results varied and it appeared that the way OC is removed from the soot is probably the most important factor of influence. The correlation between the laboratories was good as a whole but some systematic differences could be detected. Besides the different techniques to remove the organic carbon, the pretreatment of the filter by HCl (either as a vapor or as a solution) to remove the inorganic carbonates (potential interference sources), is probably also a significant factor of influence in the dispersion of the results between laboratories. It is not yet clear from these results whether the "environmental" laboratories give different results from the "occupational" laboratories, but it is clear that their objectives differ since for the "environmentalists", EC is not a specific marker of diesel emmissions, in contrast to the "occupationalists"., Conclusion: It can be concluded that, although significant differences exist between laboratories they can be attributed mainly to the narrow distribution of the results within a single laboratory, and that the overall agreement of the results for EC and TC is fairly good. These results obtained with pure diesel engine emissions, should be complemented by field samples, but they have already achieved relevant findings in the performance of the procedures used to assess exposure to diesel soot.

A survey of X-ray film processing departments revealed the main airborne contaminants to be sulphur dioxide and acetic acid at concentrations of about 0.1 ppm. Glutaraldehyde was not detected either in the ambient air or in the exhaust duct from an automatic film processor. Laboratory studies confirmed sulphur dioxide and acetic acid as the main headspace constituents above working strength processing solutions but glutaraldehyde and butyraldehyde were also detected.

Methods for monitoring personal exposure to tetrachloroethylene vapour in the dry cleaning industry have been investigated. A Draeger diffusive colorimetric gas detector tube and the Delta vapour detection system manufactured by Sabre Gas Detection were both shown to be suitable for preliminary on-site monitoring. A range of short-term pumped colorimetric tubes were also shown to perform well under a range of environmental conditions. Data on laboratory evaluations and field trials are presented.

The evolution of gases during the handling of animal slurry was investigated at five sites. Particular attention was paid to the mixing and emptying operations since it is when performing these that personnel are most likely to be at risk of exposure. The main hazard was found to be high transient concentrations of hydrogen sulphide presenting in some cases a serious acute toxicity problem. Time-weighted average exposures did not generally indicate any long-term exposure risk. Other features noted were the evolution of ammonia and methane, although not at high levels, and some increase in the carbon dioxide concentration. Some reduction in oxygen concentration was measured, but generally the risk of poisoning by hydrogen sulphide was more serious than the risk of asphyxiation.

A diffusive sampling system, using a commercially available liquid sorbent badge, has been evaluated for the determination of airborne formaldehyde concentrations according to the U.K. Health and Safety Executive protocol. The results of laboratory and field experiments are presented. The results obtained using the diffusive sampler during field measurements were about 10% higher overall than those obtained using a conventional pumped sampler.

The specific target behavior of encopresis in individuals with mental retardation and developmental disorders (MR/DD) is not well-defined in the literature on encopresis or in the literature on specific interventions with MR/DD populations. The criteria for establishing a diagnosis of encopresis in individuals with MR/DD are not clear. The literature on encopresis was reviewed for two factors: (1) definitions according to three major definitional criteria of age, defecation patterns and etiology; and (2) the relationship between etiology and intervention in specific, MR/DD, and nonspecific populations. This review yielded no clinical description of encopresis in MR/DD children, no guidelines for differentiating encopresis from generalized developmental delay and no substantive treatment guidelines. Although no definitions or treatment formulations specific to MR/DD were identified, information and definitional guidelines derived from the general literature on encopresis are relevant to identification, definition and treatment issues for MR/DD populations. Interdisciplinary or team approaches are appropriate for individuals with multiple problems or developmental disorders and intractable encopresis. An interdisciplinary approach, neurodevelopmental-behavioral intervention, which was successfully employed with a 5-year-old encopretic child with multiple developmental problems, is described.

Measurements of hazardous gas concentrations in forage tower silos are described. The problems associated with sampling in silos, the identity and distribution of gases and their rate of evolution, and the effects of forced and natural ventilation are considered. Particular attention is given to conditions at times when entry to the silo by farm personnel is likely and the possibility of farmers carrying out gas measurements is discussed.