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This issue of "Tap into Learning" continues an exploration of ways computers can support student learning. As in previous issues, a constructivist learning theory is used as a framework for discussion. This issue examines the notion that social interaction plays an important part in the construction of knowledge. Articles include: "Communication: A Key to Learning,""Connections along the Flyway,""Making It Happen in Your Classroom,""What Does Constructivist Theory Tell Us? The Butterfly Project," and "Communicating on the 'Net: A Selection of Internet Sites." The following six principles that make up a framework for constructivism are outlined: "Learners bring unique prior knowledge and beliefs to a learning situation"; "Knowledge is constructed uniquely and individually, in multiple ways, through a variety of authentic tools, resources, experiences and contexts"; "Learning is both an active and a reflective process"; "Learning is developmental. We make sense of our world by assimilating, accommodating, or rejecting new information"; "Social interaction introduces multiple perspectives on learning"; and "Learning is internally controlled and mediated by the learner." (AEF)

This issue of "TAP into Learning" focuses on technology-assisted learning activities for students, in particular on those that use spreadsheets. Articles include: "Using What Learners Know"; "Activity: Grade 7 and 8 Math, Social Studies and Language Arts"; "Managing Growth: Collaborative Decision-Making in Urban Planning"; "Spreadsheets"; "Getting Started with Spreadsheets"; and "But Aren't Spreadsheets Just for Math?" A sidebar discusses John Locke's "Essay Concerning Human Understanding." The following six principles that make up a framework for constructivism are outlined: "Learners bring unique prior knowledge and beliefs to a learning situation"; "Knowledge is constructed uniquely and individually, in multiple ways, through a variety of authentic tools, resources, experiences and contexts"; "Learning is both an active and a reflective process"; "Learning is developmental. We make sense of our world by assimilating, accommodating, or rejecting new information"; "Social interaction introduces multiple perspectives on learning"; and "Learning is internally controlled and mediated by the learner." (AEF)

Immune aplastic anemia (AA) features somatic loss of HLA class I allele expression on bone marrow cells, consistent with a mechanism of escape from T-cell–mediated destruction of hematopoietic stem and progenitor cells. The clinical significance of HLA abnormalities has not been well characterized. We examined the somatic loss of HLA class I alleles and correlated HLA loss and mutation-associated HLA genotypes with clinical presentation and outcomes after immunosuppressive therapy in 544 AA patients. HLA class I allele loss was detected in 92 (22%) of the 412 patients tested, in whom there were 393 somatic HLA gene mutations and 40 instances of loss of heterozygosity. Most frequently affected was HLA-B*14:02, followed by HLA-A*02:01, HLA-B*40:02, HLA-B*08:01, and HLA-B*07:02. HLA-B*14:02, HLA-B*40:02, and HLA-B*07:02 were also overrepresented in AA. High-risk clonal evolution was correlated with HLA loss, HLA-B*14:02 genotype, and older age, which yielded a valid prediction model. In 2 patients, we traced monosomy 7 clonal evolution from preexisting clones harboring somatic mutations in HLA-A*02:01 and HLA-B*40:02. Loss of HLA-B*40:02 correlated with higher blood counts. HLA-B*07:02 and HLA-B*40:01 genotypes and their loss correlated with late-onset of AA. Our results suggest the presence of specific immune mechanisms of molecular pathogenesis with clinical implications. HLA genotyping and screening for HLA loss may be of value in the management of immune AA. This study was registered at clinicaltrials.gov as NCT00001964, NCT00061360, NCT00195624, NCT00260689, NCT00944749, NCT01193283, and NCT01623167.

Primitive quiescent CD34+ chronic myeloid leukemia (CML) cells are more biologically resistant to tyrosine kinase inhibitors than their cycling counterparts; however, graft-versus-leukemia (GVL) effects after allogeneic stem cell transplantation (SCT) probably eliminate even these quiescent cells in long-term surviving CML transplant recipients. We studied the progeny of CD34+ cells from CML patients before SCT, which were cultured 4 days in serum-free media with hematopoietic growth factors. BCR-ABL expression was similar in both cycling and quiescent noncycling CD34+ populations. Quiescent CD34+ cells from CML patients were less susceptible than their cycling CD34+ and CD34− counterparts to lysis by natural killer (NK) cells from their HLA-identical sibling donors. Compared with cycling populations, quiescent CD34+ CML cells had higher surface expression of tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) receptors DR4 and DR5. Bortezomib up-regulated TRAIL receptor expression on quiescent CD34+ CML cells, and further enhanced their susceptibility to cytotoxicity by in vitro expanded donor NK cells. These results suggest that donor-derived NK cell–mediated GVL effects may be improved by sensitizing residual quiescent CML cells to NK-cell cytotoxicity after SCT. Such treatment, as an adjunct to donor lymphocyte infusions and pharmacologic therapy, may reduce the risk of relapse in CML patients who require treatment by SCT.

Primitive quiescent CD34+chronic myeloid leukemia (CML) cells are more biologically resistant to tyrosine kinase inhibitors than their cycling counterparts; however, graft-versus-leukemia (GVL) effects after allogeneic stem cell transplantation (SCT) probably eliminate even these quiescent cells in long-term surviving CML transplant recipients. We studied the progeny of CD34+cells from CML patients before SCT, which were cultured 4 days in serum-free media with hematopoietic growth factors. BCR-ABLexpression was similar in both cycling and quiescent noncycling CD34+populations. Quiescent CD34+cells from CML patients were less susceptible than their cycling CD34+and CD34−counterparts to lysis by natural killer (NK) cells from their HLA-identical sibling donors. Compared with cycling populations, quiescent CD34+CML cells had higher surface expression of tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) receptors DR4 and DR5. Bortezomib up-regulated TRAIL receptor expression on quiescent CD34+CML cells, and further enhanced their susceptibility to cytotoxicity by in vitro expanded donor NK cells. These results suggest that donor-derived NK cell–mediated GVL effects may be improved by sensitizing residual quiescent CML cells to NK-cell cytotoxicity after SCT. Such treatment, as an adjunct to donor lymphocyte infusions and pharmacologic therapy, may reduce the risk of relapse in CML patients who require treatment by SCT.

This study describes a novel application of HLAMatchmaker to determine platelet compatibility in 16 alloimmunized patients with aplastic anemia refractory to random donor platelet transfusions. HLAMatchmaker is a software algorithm that predicts HLA compatibility by identifying immunogenic epitopes represented by amino acid triplets in antibody-accessible regions of human leukocyte antigen (HLA) molecules and determines the number of triplet mismatches (TMMs) and highly immunogenic triplet mismatches (HIMMs). Corrected count increments (CCIs) and molecular HLA typing were available for 523 transfusions. Conventional compatibility assessment based on cross-reactive group (CREG) determination was not predictive of transfusion outcome. Low HIMMs and TMMs numbers were associated with a higher likelihood of satisfactory (CCIs ≥ 8) compared with unsatisfactory (CCIs < 8) outcomes (median HIMMs = 4 vs 6, p2value < .001; median TMMs = 11 vs 13, p2value < .001). Although receiver operator characteristic curves revealed that HIMMs or TMMs number are not powerful predictors of individual transfusion outcome, a threshold of at least 3 HIMMs or at least 9 TMMs appeared to be associated with successful transfusions. Triplet-matched transfusions were successful, regardless of CREG matching. Our data validate HLAMatchmaker for platelet transfusions and demonstrate its potential to refine and expand donor selection for HLA-alloimmunized patients.

The authors in this article state that local government restructuring in South Africa brought together 16 libraries from three different administrations to form the former South Peninsula Municipality (SPM) Library Service. In 2000, the SPM libraries participated in Competency-based Performance Management training. A task team representing the various libraries of the SPM undertook a fact-finding exercise. The results were revealing. It found a fragmented unit that was apparently under-resourced and over-committed, with demoralized staff operating almost exclusively in reactive mode, without an adequate leadership structure or a collective vision for the service, confused as to roles and responsibilities, with no adequate performance measurement in place, three different systems operating, and a number of working condition problems. The article informs that after a series of workshops with staffs, the mandate and responsibilities of the interest groups were identified.

Aims and background The genetic complexity of the human major histocompatibility complex (MHC) has required the development of various molecular typing methods. The purpose of this paper is to compare the results of two of these molecular methods: sequenced based typing (SBT) and polymerase chain reaction (PCR) using sequence specific primers (PCR-SSP).Methods The SBT method described utilizes an ABI Prism® 3700 DNA Analyzer, which has been designed fro high throughput production of sequence data through highly automated operation with significant walk-away time. The ABI Prism® 3700 DNA Analyzer is a 96-capillary electrophoresis instrument with the capability of running four 96-well plates black to back in a sixteen-hour period. Potentially, data from this machine can produce Class I sequences for A or B loci for 64 samples in this time frame. The SBT method encompassed exons 2, 3, and 4 with forward and reverse sequence orientation reactions using the PE Biosystems HLA-A and HLA-B Sequenced Based Typing Kits (PE Applied Biopsystems/Perkin-Elmer, Foster City, CA, USA). Most SBT methods previously employed only gather data from exons 2 and 3 which distinguishes most of the polymorphism necessary to identify the majority of alleles in the HLA region. However, in an effort to discern numerous null alleles in the HLA region, exon 4 data is also included. The PCR-SSP method utilized consists of one 96 well tray, with 95 primer mixes and one negative control, per sample designed to produce an intermediate/high resolution HLA-A, B typing.Results Data from one 96-well capillary run on the ABI Prism® 3700 DNA Analyzer, which consists of results from 16 samples for HLA-A or HLA-B loci, was compared to data derived from sixteen HLA-A and HLA-B PCR-SSP typings. 75% of loci tested achieved a higher resolution HLA typing by the SBT method.Discussion The ability to provide allele level HLA typing results can have significant functional implications for the bone marrow transplant community and numerous vaccine studies.

When administered as single agents, both interferon gamma (IFN-γ) and interleukin 6 (IL-6) significantly increase carcinoembryonic antigen (CEA) and HLA class I antigen expression on the surface of human colorectal tumour cells. Studies were carried out to determine whether by combining those cytokines a synergistic enhancement of CEA and HLA expression could result. The findings revealed that the administration of 20 units IFN-γ along with 1.7 ng IL-6, concentrations of each cytokine that individually induced minimal antigenic changes, together synergistically increased CEA and HLA class I as well as induced qualitative changes in HLA expression on WiDr human colon carcinoma cells. The magnitude of the synergistic increases in CEA and HLA class I expression were reminiscent of the level of antigen augmentation observed when administering 20- to 100-fold higher amounts of each cytokine as a single agent. Also, the addition of IL-6 potentiated the IFN-γ induction of HLA class II expression. The combined administration of IL-6 potentiated the IFN-γ did not have any additive or synergistic effects on the growth suppression of those tumour cells. Interestingly, utilization of specific neutralizing antibodies for type I interferons abrogated the increases of CEA and HLA expression seen with IL-6 treatment alone or in combination with IFN-γ. Moreover, reverse transcriptase/polymerase chain reaction analyses revealed a constitutive expression as well as a temporal increase of IFN-β mRNA transcripts in colon tumour cells treated with IL-6. Therefore, the findings provide indirect evidence that IFN-β production seems to play a critical role in the ability of IL-6 to upregulate antigen expression alone or in combination with IFN-γ. These findings provide insight into cytokine combinations that synergistically upregulate tumour-associated and normal HLA antigen expression on the surface of human tumour cells. Those results provide the rationale for the combined use of such cytokines to heighten tumour cell recognition in monoclonal antibody- or cell-mediated-based immunotherapeutic approaches.

We have undertaken a combined molecular and genetic analysis of the bronze (bz) locus in maize. The bz locus was isolated by a two-step approach involving sequences from the transposable elements Dissociation (Ds) and Activator (Ac). A restriction map of the bz region is presented, identifying the transcribed region and the direction of transcription. We have mapped the sites of two Ds insertions, bz-m2 (DI) and bz-m1, physically and genetically. bz-m2 (DI) is a 3.5 kb insertion in the transcribed region that appears to have arisen by an internal deletion in the 4.5 kb Ac insertion in the mutant bz-m2. bz-m1 is a 1.1 kb insert in the 5' end of the gene. By correlating the genetic and physical maps, we have been able to orient the restriction map relative to the centromere in chromosome 9 and have obtained a rough estimate of the physical length of a unit of genetic recombination in a microregion of the maize genome.

This article discusses the proceedings of the National Lawyer's Guild held in Venezuela. The delegation was guided by Lisa Sullivan who is working for the Venezuelan Information Office. The delegation participated as North Americans opposed to U.S. Intervention in Venezuela. They believe that the Venezuelan process should be allowed to unfold at its own accord, without U.S. intervention.

Introduction. Allogeneic hematopoietic stem cell transplantation using UCB is an alternative approach for pts (pts) with hematological malignancies lacking an HLA matched donor. However, for pts with severe aplastic anemia (SAA), UCB transplants are associated with delayed engraftment, high graft failure rates and poor survival. Ex-vivo expanded UCB using nicotinamide (NAM) can engraft in NOD/SCID mice, and in pilot studies in pts with hematological malignancies, results in rapid engraftment and durable hematopoiesis. Here we investigated a novel transplantation approach using NAM-expanded UCB in refractory SAA pts who lacked an HLA matched donor, hypothesizing this regimen would accelerate engraftment and immune reconstitution compared to conventional UCB transplants.

Several letters to the editor are presented in response to articles in previous issues including "Policy changes: SECNAV orders review of fitness, advancement" in the June 22, 2015 issue, "May courts-martial results announced" in the June 16, 2015 issue, and atheists.

Background:

Background:The development of HLA antibodies is responsible for 4-8% of platelet transfusion refractoriness (PTR). The presence of HLA antibodies is a clinical complication that is generally managed by the selection of products that are negative for the antigens cognate to the patient's antibodies. Often, this selection is facilitated by targeted recruitment of donors with known HLA types. However, for very broadly HLA-alloimmunized patients, compatible products may be exceedingly scarce or completely unavailable, precluding the ability to consistently provide products the patient will likely increment to and benefit from. Here we describe a case of a patient with severe aplastic anemia (platelet count chronically <1 K/uL) and platelet transfusion refractoriness secondary to broad HLA-alloimmunization (98% PRA). HLA compatible products were unavailable due to the patient's rare HLA type and breadth of antibodies, and attempts at procuring compatible products via multiple national agency searches were unsuccessful. Thus, the patient could only be supplied with HLA incompatible products, which yielded insufficient post-transfusion platelet corrected-count increments (CCI). Her course was eventually complicated by multifocal intracranial hemorrhages, necessitating ICU admission with aggressive platelet transfusion support, immune-targeting therapy consisting of IVIg, and aminocaproic acid and recombinant activated factor VIIa.

Introduction:

Several letters to the editor are presented in response to articles in previous issues, including "The Good Life" and an article about paeony, both in the November 2011 issue, as well as a letter in which the sender talks about hydroponics.

The article reports on the federal trademark registration received by the American National Lawyers Guild for the words Legal Observer (LO). The official trademark will enhance the protection of activists and their free speech rights in the face of developments. This article also discusses the challenges faced by the LOs including the government's intervention and negative reports of several organizations.

Young: GSK/Novartis: Research Funding.

No relevant conflicts of interest to declare.

Several letters to the editor are presented discussing vacation in Vietnam as well as taking pictures from the top of the Parkridge subdivision near Iroquois Park in Louisville, Kentucky.

The article focuses on three lawyers who are advocates of torture as well as on the activities being completed against them for their illegal and immoral actions in the U.S. University of California-Berkeley law professor John Yoo is faced with petition to oust him as law professor. 9th Circuit judge Jay Bybee has already received a complaint of torture and misconduct against him. Department of Defense former general counsel William Haynes, moreover, has received a complaint for misconduct.

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

Off Label Use: There is currently no FDA approved product for GVHD prevention or therapy. The GVHD prophylaxis regimens described in this study (Alemtuzumab-Cyclosporine and Tacrolimus-Methotrexate-Sirolimus) are used off-label as GVHD prophylaxis regimens in reduced-intensity allogeneic HSCT.

INTRODUCTION:

Off Label Use: BL22 and Moxetumomab Pasudotox are experimental agents for CD22+ malignancies. Pastan:NIH: Coinventor on NIH patents for BL22 and Moxetumomab, Coinventor on NIH patents for BL22 and Moxetumomab Patents & Royalties. Kreitman:NIH: Coinventor on NIH patents for BL22 and Moxetumomab, Coinventor on NIH patents for BL22 and Moxetumomab Patents & Royalties.

No relevant conflicts of interest to declare.

Abstract 4201

The article reports on the support of National Lawyers Guild (NLG) members towards the protection of indigenous occupation of a sacred Native American site in Vallejo, California. It mentions the organization Sacred Sites Protection & Rights of Indigenous Tribes (SSP&RIT) which was established against desecration of the site along with its request for legal assistance from the guild's Legal Oberservers (LOs). It also notes the successful negotation of the organization with city officials.

Wilder: NCI: Funded in part by NCI contract No. HHSN261200800001E.

Platelet refractoriness as a consequence of HLA alloimmunization complicates allogeneic hematopoietic stem cell transplantation (HSCT). Little is known regarding the affects of reduced intensity HSCT on the incidence and duration of HLA alloimmune-mediated platelet refractoriness following reduced intensity HSCT.We retrospectively studied HLA alloimmunization in 16 patients with malignant and non-malignant hematological disorders, who underwent an allogeneic HSCT from an HLA matched relative using reduced intensity, non-myeloablative conditioning. All patients received a G-CSF mobilized, lymphocyte replete, peripheral blood stem cell (PBSC) allograft following cyclophosphamide (120 mg/kg) and fludarabine (125 mg/m2) based conditioning (+/− equine ATG 40 mg/kg × 4 days) with cyclosporine (CSA; beginning on day −4) and IV methotrexate (5 mg/m2 × 3 days) given as GVHD prophylaxis. Eight patients known to be HLA alloimmunized pre-transplant were compared to 8 control patients who were HLA alloantibody negative pre-transplant. Stored patients' serum samples from pre-transplant (day -30) and post-transplant intervals (days +30, +60, +100, +180, and ≥ 365) were analyzed for the presence of IgG antibodies to HLA class I antigens using a membrane-independent solid phase assay involving color-coded microbeads coated with HLA antigens and analysis with a flow analyzer (LABScan 100; One Lambda, Canoga Park, CA). Panel reactive antibody (PRA) and mean fluorescent intensity (MFI) were analyzed for all samples to measure HLA antibody strength and specificity, respectively. HLA alloantibodies were analyzed and compared with the degree of donor myeloid and T-cell engraftment measured on post-transplant blood samples by PCR of short tandem repeats (STRs).Among the 8 alloimmunized patients who required HLA matched platelets pre-transplant, the median time until HLA antibodies disappeared was 100 days post HSCT. Remarkably, among these patients, 3/8 (37%) had HLA class I antibodies detectable for more than 100 days post-transplant including one patient who continued to show high level alloreactivity greater than 1 year after HSCT. Among the 8 control patients who tested negative for HLA alloantibodies pre-transplant, 3 acquired HLA alloantibodies after HSCT which were first detected at day +30 in all 3 cases. In 2 of the 3 cases, the donors for these patients were found to have pre-existing HLA antibodies of equivalent specificity to those found in the patient post-transplant suggesting patient acquisition of HLA antibodies was mediated by passenger donor lymphocytes that were transplanted in the allograft. Overall, HLA antibodies were detectable for more than 100 days after transplantation in 6/16 (37%) patients analyzed with 3/16 (18%) having alloantibodies detectable for more than 1 year post-transplant despite chimerism being 100% donor in myeloid and T cell lineages.Prolonged production of HLA alloantibodies leading to platelet refractoriness can occur following reduced intensity allogeneic HSCT and may persist even after full donor myeloid and T-cell chimerism have been achieved. Remarkably, we observed that transplantation of passenger donor lymphocytes can result in de novo HLA alloimmunization, complicating post-transplant transfusion management. Screening patients and their donors for HLA antibodies before HSCT would identify the majority of subjects at risk for alloimmune-mediated platelet refractoriness after transplantation. This new screening strategy would not only assist in transfusion support after allogeneic HSCT but could also play a role in the decision analysis for selecting optimal stem cell donors.No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

The article reports on U.S. President George Bush Administration's use of torture that was disputed by the National Lawyers Guild (NLG). NLG issued a press release against law professor John Yoo and former council David Addington for an equivocal cause agitating the unilateral power of the president to order torture of a suspect. Yoo, having specified the conduct at issue, is planned by Berkeley NLG groups with charges pursuant to his involvement in war crimes and crimes against humanity.

The article reports on the move by Jeff Heaton, a member of the National Lawyers Guild, on supporting Iraq pro-war faction memorial by filing a sign ordinance. It was revealed that the Lafayette sign ordinance, like most sign ordinance, relies on the concept of aesthetics to limit speech. In addition, the proposed new sign ordinance would have exempted existing non-compliant signs, but provided that any change to an existing non-compliant sign would make the sign "illegal."

Presents a letter to the editor discussing gardening tips.