bluuebunny/crossref_metadata_2025 huggingface dataset:
Protein kinase B (c-akt) regulates hematopoietic lineage choice decisions during myelopoiesis
Authors: Buitenhuis Miranda , Verhagen Liesbeth P. , van Deutekom Hanneke W. M. , Castor Anders , Verploegen Sandra , Koenderman Leo , Jacobsen Sten-Eirik W. , Coffer Paul J.
Published: September 2007

Hematopoiesis is a highly regulated process resulting in the formation of all blood lineages. Aberrant regulation of phosphatidylinositol-3-kinase (PI3K) signaling has been observed in hematopoietic malignancies, suggesting that regulated PI3K signaling is critical for regulation of blood cell production. An ex vivo differentiation system was used to investigate the role of PI3K and its downstream effector, protein kinase B (PKB/c-akt) in myelopoiesis. PI3K activity was essential for hematopoietic progenitor survival. High PKB activity was found to promote neutrophil and monocyte development, while, conversely, reduction of PKB activity was required to induce optimal eosinophil differentiation. In addition, transplantation of β2-microglobulin (−/−) NOD/SCID mice with CD34+ cells ectopically expressing constitutively active PKB resulted in enhanced neutrophil and monocyte development, whereas ectopic expression of dominant-negative PKB induced eosinophil development in vivo. Inhibitory phosphorylation of C/EBPα on Thr222/226 was abrogated upon PKB activation in hematopoietic progenitors. Ectopic expression of a nonphosphorylatable C/EBPα mutant inhibited eosinophil differentiation ex vivo, whereas neutrophil development was induced, demonstrating the importance of PKB-mediated C/EBPα phosphorylation in regulation of granulopoiesis. These results identify an important novel role for PKB in regulation of cell fate choices during hematopoietic lineage commitment.
Fertility in male patients with advanced Hodgkin lymphoma treated with BEACOPP: a report of the German Hodgkin Study Group (GHSG)
Authors: Sieniawski Michal , Reineke Thorsten , Nogova Lucia , Josting Andreas , Pfistner Beate , Diehl Volker , Engert Andreas
Published: September 2007

To date, there is little information on the impact of more aggressive treatment regimen such as BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone) on the fertility of male patients with Hodgkin lymphoma (HL). We evaluated the impact of BEACOPP regimen on fertility status in 38 male patients with advanced-stage HL enrolled into trials of the German Hodgkin Study Group (GHSG). Before treatment, 6 (23%) patients had normozoospermia and 20 (77%) patients had dysspermia. After treatment, 34 (89%) patients had azoospermia, 4 (11%) had other dysspermia, and no patients had normozoospermia. There was no difference in azoospermia rate between patients treated with BEACOPP baseline and those given BEACOPP escalated (93% vs 87%, respectively; P > .999). After treatment, most of patients (93%) had abnormal values of follicle-stimulating hormone, whereas the number of patients with abnormal levels of testosterone and luteinizing hormone was less pronounced—57% and 21%, respectively. In univariate analysis, none of the evaluated risk factors (ie, age, clinical stage, elevated erythrocyte sedimentation rate, B symptoms, large mediastinal mass, extranodal disease, and 3 or more lymph nodes) was statistically significant. Male patients with HL are at high risk of infertility after treatment with BEACOPP.
A Stat5b transgene is capable of inducing CD8+ lymphoblastic lymphoma in the absence of normal TCR/MHC signaling
Authors: Bessette Katherine , Lang Mark L. , Fava Roy A. , Grundy Martin , Heinen Jennifer , Horne Laurie , Spolski Rosanne , Al-Shami Amin , Morse Herbert C. , Leonard Warren J. , Kelly John A.
Published: September 2007

Stat5 proteins are critical signaling molecules activated by many cytokines. Within the immune system, Stat5 plays important roles related to the development of thymocytes and proliferation of T cells. Stat5 has been implicated in malignant transformation, and moreover, the activated tyrosine phosphorylated form of Stat5 is frequently observed in human lymphomas. We previously demonstrated the oncogenic potential of Stat5, with thymic lymphoblastic lymphomas developing in a significant proportion of transgenic (TG) mice overexpressing Stat5a or Stat5b in lymphocytes. In addition, immunization or expression of a T-cell receptor (TCR) transgene augmented the rate of tumor formation. Here, we investigate the mechanism of Stat5-mediated lymphomagenesis by exploring the contributions of major histocompatibility complex (MHC)/TCR and pre-TCR signals. We present data demonstrating that Stat5b TG mice unexpectedly develop CD8+ lymphoma even in the absence of either pre-TCR signaling or normal thymic selection. Indeed, acceleration of Stat5b transgene-mediated lymphoma occurred on TCRα−/− and pre-TCRα−/− backgrounds. In light of these data, we propose a model in which alterations in T-cell development at the double-negative/double-positive (DN/DP) stages cooperate with cytokine-mediated pathways in immature thymocytes to give rise to lymphoblastic T-cell lymphomas in Stat5b TG mice.
Molecular allelokaryotyping of pediatric acute lymphoblastic leukemias by high-resolution single nucleotide polymorphism oligonucleotide genomic microarray
Authors: Kawamata Norihiko , Ogawa Seishi , Zimmermann Martin , Kato Motohiro , Sanada Masashi , Hemminki Kari , Yamatomo Go , Nannya Yasuhito , Koehler Rolf , Flohr Thomas , Miller Carl W. , Harbott Jochen , Ludwig Wolf-Dieter , Stanulla Martin , Schrappe Martin , Bartram Claus R. , Koeffler H. Phillip
Published: September 2007

Pediatric acute lymphoblastic leukemia (ALL) is a malignant disease resulting from accumulation of genetic alterations. A robust technology, single nucleotide polymorphism oligonucleotide genomic microarray (SNP-chip) in concert with bioinformatics offers the opportunity to discover the genetic lesions associated with ALL. We examined 399 pediatric ALL samples and their matched remission marrows at 50 000/250 000 SNP sites using an SNP-chip platform. Correlations between genetic abnormalities and clinical features were examined. Three common genetic alterations were found: deletion of ETV6, deletion of p16INK4A, and hyperdiploidy, as well as a number of novel recurrent genetic alterations. Uniparental disomy (UPD) was a frequent event, especially affecting chromosome 9. A cohort of children with hyperdiploid ALL without gain of chromosomes 17 and 18 had a poor prognosis. Molecular allelokaryotyping is a robust tool to define small genetic abnormalities including UPD, which is usually overlooked by standard methods. This technique was able to detect subgroups with a poor prognosis based on their genetic status.
Treatment-related risk factors for premature menopause following Hodgkin lymphoma
Authors: De Bruin Marie L. , Huisbrink Jeannine , Hauptmann Michael , Kuenen Marianne A. , Ouwens Gabey M. , van't Veer Mars B. , Aleman Berthe M. P. , van Leeuwen Flora E.
Published: September 2007

We conducted a cohort-study among 518 female 5-year Hodgkin lymphoma (HL) survivors, aged 14 to 40 years (median: 25 years) at treatment (1965-1995). Multivariable Cox regression was used to quantify treatment effects on risk of premature menopause, defined as cessation of menses before age 40 years. After a median follow up of 9.4 years, 97 women had reached menopause before age 40 years. Chemotherapy was associated with a 12.3-fold increased risk of premature menopause compared with radiotherapy alone. Treatment with MOPP (mechlorethamine, vincristine, procarbazine, prednisone)/ABV (doxorubicine, bleomycine, vinblastine) significantly increased the risk of premature menopause (hazard ratio [HR]: 2.9), although to a lesser extent than MOPP treatment (HR: 5.7). Alkylating agents, especially procarbazine (HR: 8.1) and cyclophosphamide (HR: 3.5), showed the strongest associations. Ten years after treatment, the actuarial risk of premature menopause was 64% after high cumulative doses (> 8.4 g/m2) and 15% after low doses (≤ 4.2 g/m2) of procarbazine. The cumulative risk of menopause at age 40 years did not differ much according to age, but time to premature menopause was much longer in women treated at early ages. As long as alkylating agents will be used for curing HL, premature menopause will remain a frequent adverse treatment effect, with various clinical implications.
The viral interferon-regulatory factor-3 is required for the survival of KSHV-infected primary effusion lymphoma cells
Authors: Wies Effi , Mori Yasuko , Hahn Alexander , Kremmer Elisabeth , Stürzl Michael , Fleckenstein Bernhard , Neipel Frank
Published: September 2007

Human herpesvirus-8 (HHV-8), also known as Kaposi sarcoma–associated herpesvirus (KSHV), is etiologically linked to primary effusion lymphoma (PEL). At least 10 KSHV-encoded proteins with potential roles in KSHV-associated neoplasia have been identified. However, with few exceptions, these putative oncogenes were analyzed in heterologous systems only using overexpression of single genes. Thus, the pathogenetic relevance of most of these putative oncogenes remains essentially unclear. We used RNA interference (RNAi) to knock down the expression of several KSHV genes in cultured PEL cells carrying the KSHV genome. The viral interferon-regulatory factor-3 (vIRF-3) was found to be required for proliferation and survival of cultured PEL cells. Knock-down of vIRF-3 expression by various RNAi approaches unequivocally resulted in reduced proliferation and increased activity of caspase-3 and/or caspase-7. Thus, vIRF-3 can be seen as a bona fide oncogene of KSHV-associated lymphoma. Surprisingly, although the related Epstein-Barr virus (EBV) is usually sufficient to immortalize human B lymphocytes, silencing of vIRF-3 reduced the viability of both EBV− and EBV+ PEL cells. This suggests that KSHV is the driving force in the pathogenesis of PEL.
Lack of iNKT cells in patients with combined immune deficiency due to hypomorphic RAG mutations
Authors: Matangkasombut Ponpan , Pichavant Muriel , Saez Doris E. , Giliani Silvia , Mazzolari Evelina , Finocchi Andrea , Villa Anna , Sobacchi Cristina , Cortes Patricia , Umetsu Dale T. , Notarangelo Luigi D.
Published: September 2007

Hypomorphic mutations of the RAG genes in humans are associated with a spectrum of clinical and immunologic presentations that range from T− B− severe combined immune deficiency (SCID) to Omenn syndrome. In most cases, residual V(D)J recombination activity allows for development of few T-cell clones, which expand in the periphery and infiltrate target organs, resulting in tissue damage. Invariant natural killer T (iNKT) cells play an important immunoregulatory role and have been associated with protection against autoimmunity. We now report on 5 unrelated cases of combined immune deficiency due to hypomorphic RAG mutations, and demonstrate the absence of iNKT cells in all 5 patients. These findings suggest that lack of this important immunoregulatory cell population may contribute to the pathophysiology of Omenn syndrome.
Thrombospondin-1 stimulates platelet aggregation by blocking the antithrombotic activity of nitric oxide/cGMP signaling
Authors: Isenberg Jeff S. , Romeo Martin J. , Yu Christine , Yu Christine K. , Nghiem Khauh , Monsale Jude , Rick Margaret E. , Wink David A. , Frazier William A. , Roberts David D.
Published: September 2007

Platelet α-granules constitute the major rapidly releasable reservoir of thrombospondin-1 in higher animals. Although some fragments and peptides derived from thrombospondin-1 stimulate or inhibit platelet aggregation, its physiologic function in platelets has remained elusive. We now show that endogenous thrombospondin-1 is necessary for platelet aggregation in vitro in the presence of physiologic levels of nitric oxide (NO). Exogenous NO or elevation of cGMP delays thrombin-induced platelet aggregation under high shear and static conditions, and exogenous thrombospondin-1 reverses this delay. Thrombospondin-1–null murine platelets fail to aggregate in response to thrombin in the presence of exogenous NO or 8Br-cGMP. At physiologic concentrations of the NO synthase substrate arginine, thrombospondin-1–null platelets have elevated basal cGMP. Ligation of CD36 or CD47 is sufficient to block NO-induced cGMP accumulation and mimic the effect of thrombospondin-1 on aggregation. Exogenous thrombospondin-1 also reverses the suppression by NO of αIIb/β3 integrin–mediated platelet adhesion on immobilized fibrinogen, mediated in part by increased GTP loading of Rap1. Thrombospondin-1 also inhibits cGMP-mediated activation of cGMP-dependent protein kinase and thereby prevents phosphorylation of VASP. Thus, release of thrombospondin-1 from α-granules during activation provides positive feedback to promote efficient platelet aggregation and adhesion by overcoming the antithrombotic activity of physiologic NO.
High TCL1 expression and intact T-cell receptor signaling define a hyperproliferative subset of T-cell prolymphocytic leukemia
Authors: Herling Marco , Patel Kaushali A. , Teitell Michael A. , Konopleva Marina , Ravandi Farhad , Kobayashi Ryuji , Jones Dan
Published: September 2007

The T-cell leukemia 1 (TCL1) oncoprotein is overexpressed by chromosomal rearrangement in the majority of cases of T-cell prolymphocytic leukemia (T-PLL). In vitro, TCL1 can modulate the activity of the serine-threonine kinase AKT, a downstream effector of T-cell receptor (TCR) signaling. In a series of 86 T-PLL tumors, we show that expression of TCR, and levels of TCL1 and activated AKT are adverse prognostic markers. High-level TCL1 in TCR-expressing T-PLL is associated with higher presenting white blood cell counts, faster tumor cell doubling, and enhanced in vitro growth response to TCR engagement. In primary tumors and TCL1-transfected T-cell lines, TCR engagement leads to rapid recruitment of TCL1 and AKT to transient membrane activation complexes that include TCR-associated tyrosine kinases, including LCK. Pharmacologic inhibition of AKT activation alters the localization, stability, and levels of these transient TCL1-AKT complexes and reduces tumor cell growth. Experimental introduction and knockdown of TCL1 influence the kinetics and strength of TCR-mediated AKT activation. We propose that in T-PLL, TCL1 represents a highly regulated, targetable modulator of TCR-mediated AKT growth signaling.
Combining classifiers to predict gene function in Arabidopsis thaliana using large-scale gene expression measurements
Authors: Lan Hui , Carson Rachel , Provart Nicholas J , Bonner Anthony J
Published: September 2007

Background

Arabidopsis thalianais the model species of current plant genomic research with a genome size of 125 Mb and approximately 28,000 genes. The function of half of these genes is currently unknown. The purpose of this study is to infer gene function in Arabidopsis using machine-learning algorithms applied to large-scale gene expression data sets, with the goal of identifying genes that are potentially involved in plant response to abiotic stress.

Results

Using in house and publicly available data, we assembled a large set of gene expression measurements forA. thaliana. Using those genes of known function, we first evaluated and compared the ability of basic machine-learning algorithms to predict which genes respond to stress. Predictive accuracy was measured using ROC50and precision curves derived through cross validation. To improve accuracy, we developed a method for combining these classifiers using a weighted-voting scheme. The combined classifier was then trained on genes of known function and applied to genes of unknown function, identifying genes that potentially respond to stress. Visual evidence corroborating the predictions was obtained using electronic Northern analysis. Three of the predicted genes were chosen for biological validation. Gene knockout experiments confirmed that all three are involved in a variety of stress responses. The biological analysis of one of these genes (At1g16850) is presented here, where it is shown to be necessary for the normal response to temperature and NaCl.

Conclusion

Supervised learning methods applied to large-scale gene expression measurements can be used to predict gene function. However, the ability of basic learning methods to predict stress response varies widely and depends heavily on how much dimensionality reduction is used. Our method of combining classifiers can improve the accuracy of such predictions – in this case, predictions of genes involved in stress response in plants – and it effectively chooses the appropriate amount of dimensionality reduction automatically. The method provides a useful means of identifying genes inA. thalianathat potentially respond to stress, and we expect it would be useful in other organisms and for other gene functions.
Unravelling the hidden heterogeneities of diffuse large B-cell lymphoma based on coupled two-way clustering
Authors: Zhang Wei , Li Li , Li Xia , Jiang Wei , Huo Jianmin , Wang Yadong , Lin Meihua , Rao Shaoqi
Published: September 2007

Background

It becomes increasingly clear that our current taxonomy of clinical phenotypes is mixed with molecular heterogeneity. Of vital importance for refined clinical practice and improved intervention strategies is to define the hidden molecular distinct diseases using modern large-scale genomic approaches. Microarray omics technology has provided a powerful way to dissect hidden genetic heterogeneity of complex diseases. The aim of this study was thus to develop a bioinformatics approach to seek the transcriptional features leading to the hidden subtyping of a complex clinical phenotype. The basic strategy of the proposed method was to iteratively partition in two ways sample and feature space with super-paramagnetic clustering technique and to seek for hard and robust gene clusters that lead to a natural partition of disease samples and that have the highest functionally conceptual consensus evaluated with Gene Ontology.

Results

We applied the proposed method to two publicly available microarray datasets of diffuse large B-cell lymphoma (DLBCL), a notoriously heterogeneous phenotype. A feature subset of 30 genes (38 probes) derived from analysis of the first dataset consisting of 4026 genes and 42 DLBCL samples identified three categories of patients with very different five-year overall survival rates (70.59%, 44.44% and 14.29% respectively;p= 0.0017). Analysis of the second dataset consisting of 7129 genes and 58 DLBCL samples revealed a feature subset of 13 genes (16 probes) that not only replicated the findings of the important DLBCL genes (e.g.JAW1andBCL7A), but also identified three clinically similar subtypes (with 5-year overall survival rates of 63.13%, 34.92% and 15.38% respectively;p= 0.0009) to those identified in the first dataset. Finally, we built a multivariate Cox proportional-hazards prediction model for each feature subset and definedJAW1as one of the most significant predictor (p= 0.005 and 0.014; hazard ratios = 0.02 and 0.03, respectively for two datasets) for both DLBCL cohorts under study.

Conclusion

Our results showed that the proposed algorithm is a promising computational strategy for peeling off the hidden genetic heterogeneity based on transcriptionally profiling disease samples, which may lead to an improved diagnosis and treatment of cancers.
Leishmania amazonensisinfection impairs differentiation and function of human dendritic cells
Authors: Favali Cecilia , Tavares Natália , Clarêncio Jorge , Barral Aldina , Barral-Netto Manoel , Brodskyn Claudia
Published: September 2007

Dendritic cells (DCs) are of utmost importance in initiating an immune response and may also function as targets for pathogens. The presence of pathogens inside DCs is likely to impair their functions and thus, influence immune responses. In the present report, we evaluated the impact of the presence of Leishmania amazonensis during differentiation and maturation of human monocyte-derived DCs. The presence of live L. amazonensis parasites during DC differentiation led to a significant decrease in CD80 (92%) and CD1a (56%) expression and an increase in CD86 (56%) cell surface expression. Phenotypic changes were accompanied by a lower secretion of IL-6, observed after 6 days of DC differentiation in the presence of L. amazonensis. DCs differentiated in the presence of L. amazonensis were used as APC in an autologous coculture, and lower amounts of IFN-γ were obtained compared with control DCs differentiated in the absence of parasites. The effect of heat-killed parasites, but not of Leishmania antigen, during DC differentiation and maturation was similar to that observed with viable parasites. During maturation, the presence of live L. amazonensis parasites, but not of soluble Leishmania antigen, led to a decrease in IL-6 and IL-10 production. In this way, we observed that the parasite is able to abrogate full DC differentiation, causing a delay in the immune response and likely, favoring its establishment in human hosts.
Ginsenoside Re Reduces Insulin Resistance through Inhibition of c-Jun NH2-Terminal Kinase and Nuclear Factor-κB
Authors: Zhang Zhiguo , Li Xiaoying , Lv Wenshan , Yang Yisheng , Gao Hong , Yang Jun , Shen Yun , Ning Guang
Published: September 2007

Ginsenoside Re (Re), a compound derived from Panax ginseng, shows an antidiabetic effect. However, the molecular basis of its action remains unknown. We investigated insulin signaling and the antiinflammatory effect by Re in 3T3-L1 adipocytes and in high-fat diet (HFD) rats to dissect its anti-hyperglycemic mechanism. Glucose uptake was measured in 3T3-L1 cells and glucose infusion rate determined by clamp in HFD rats. The insulin signaling cascade, including insulin receptor (IR) β-subunit, IR substrate-1, phosphatidylinositol 3-kinase, Akt and Akt substrate of 160 kDa, and glucose transporter-4 translocation are examined. Furthermore, c-Jun NH2-terminal kinase (JNK), MAPK, and nuclear factor (NF)-κB signaling cascades were also assessed. The results show Re increases glucose uptake in 3T3-L1 cells and glucose infusion rate in HFD rats. The activation of insulin signaling by Re is initiated at IR substrate-1 and further passes on through phosphatidylinositol 3-kinase and downstream signaling cascades. Moreover, Re demonstrates an impressive suppression of JNK and NF-κB activation and inhibitor of NF-κBα degradation. In conclusion, Re reduces insulin resistance in 3T3-L1 adipocytes and HFD rats through inhibition of JNK and NF-κB activation.
CR6-Interacting Factor 1 Represses the Transactivation of Androgen Receptor by Direct Interaction
Authors: Suh Ji Ho , Shong Minho , Choi Hueng-Sik , Lee Keesook
Published: September 2007

CR6-interacting factor 1 (CRIF1) was previously identified as a nuclear protein that interacts with members of the Gadd45 family and plays a role as a negative regulator in cell growth. However, the nuclear function of CRIF1 remains largely unknown. In this study, we demonstrate that CRIF1 acts as a novel corepressor of the androgen receptor (AR) in prostatic cells. Transient transfection studies show that CRIF1 specifically represses AR transcriptional activation of target promoters in a dose-dependent manner. Additionally, CRIF1 is recruited with AR to the endogenous AR target promoters. In vivo and in vitro protein interaction assays reveal that CRIF1 directly interacts with AR via the activation function-1 domain of AR. Interestingly, both the N-terminal and C-terminal half-regions of CRIF1 are independently capable of interacting with and repressing the transactivation of AR. CRIF1 represses AR transactivation through competition with AR coactivators. In addition, the CRIF1-mediated inhibition of AR transactivation involves the recruitment of histone deacetylase 4. Down-regulation of CRIF1 by small interfering RNA increases the transactivation of AR and the mRNA level of the AR target gene prostate-specific antigen, whereas the overexpression of CRIF1 decreases the prostate-specific antigen mRNA level. Finally, the overexpression of CRIF1 inhibits the androgen-induced proliferation and cell cycle progression of prostate cancer cells. Taken together, these results suggest that CRIF1 acts as an AR corepressor and may play an important role in the regulation of AR-positive growth of prostate cancer.
Influence of ScaI and Natriuretic Peptide (NP) Clearance Receptor Polymorphisms of the NP System on NP Concentration in Chronic Heart Failure
Authors: Vassalle Cristina , Andreassi Maria Grazia , Prontera Concetta , Fontana Marianna , Zyw Luc , Passino Claudio , Emdin Michele
Published: September 2007

Background: Genetic variants related to the natriuretic peptide (NP) system [ScaI mutated allele (A1) of the atrial NP (ANP) gene and the C variant of the natriuretic peptide clearance receptor (NPRC) gene] have been identified as independent risk factors for cardiovascular morbidity and mortality. Despite the importance of NPs in heart failure (HF), the role of these polymorphisms in HF has not been evaluated.

Methods: We screened 124 HF patients [mean (SD), age 66 (12) years, 100 men, ejection fraction 32% (10%), New York Heart Association (NYHA) class I–II 65, III–IV 59] for NP concentrations [ANP, brain NP (BNP) and amino-terminal pro-BNP (NT-proBNP)] and for the ScaI and NPRC variants.

Results: ScaI polymorphism had no effect on NP concentration in the NYHA I–II subgroup. Conversely, in severe HF, A1 carriers had higher ANP (P ≤0.05), BNP (P <0.01), and NT-proBNP (P <0.01) than A2A2 patients. After multivariate adjustment, A1 presence remained an independent predictor for increased NP. Regarding NPRC polymorphism in mild HF, higher ANP (P <0.05) and BNP (P <0.05) were observed in CC than A allele carriers. After multivariate adjustment, however, this association did not remain significant. In severe HF, the NPRC polymorphism had no effect on NP.

Conclusions: The ScaI polymorphism of the ANP gene might be an important additive genetic factor influencing neurohormonal activation and disease progression in severe HF. The NPRC polymorphism is not an independent determinant of NP concentration in HF.
Applications of Nanobiotechnology in Clinical Diagnostics
Authors: Jain Kewal K
Published: September 2007

Background: Nanobiotechnologies are being applied to molecular diagnostics and several technologies are in development.

Methods: This review describes nanobiotechnologies that are already incorporated in molecular diagnostics or have potential applications in clinical diagnosis. Selected promising technologies from published literature as well as some technologies that are in commercial development but have not been reported are included.

Results: Nanotechnologies enable diagnosis at the single-cell and molecule levels, and some can be incorporated in current molecular diagnostic methods, such as biochips. Nanoparticles, such as gold nanoparticles and quantum dots, are the most widely used, but various other nanotechnological devices for manipulation at the nanoscale as well as nanobiosensors are also promising for potential clinical applications.

Conclusions: Nanotechnologies will extend the limits of current molecular diagnostics and enable point-of-care diagnostics, integration of diagnostics with therapeutics, and development of personalized medicine. Although the potential diagnostic applications are unlimited, the most important current applications are foreseen in the areas of biomarker discovery, cancer diagnosis, and detection of infectious microorganisms. Safety studies are needed for in vivo use. Because of its close interrelationships with other technologies, nanobiotechnology in clinical diagnosis will play an important role in the development of nanomedicine in the future.
Methylation-Sensitive High-Resolution Melting-Curve Analysis of the SNRPN Gene as a Diagnostic Screen for Prader-Willi and Angelman Syndromes
Authors: White Helen E , Hall Victoria J , Cross Nicholas CP
Published: September 2007

Background: Angelman syndrome (AS) and Prader-Willi syndrome (PWS) are 2 distinct neurodevelopmental disorders caused primarily by deficiency of specific parental contributions at an imprinted domain within the chromosomal region 15q11.2–13. Lack of paternal contribution results in PWS either by paternal deletion (approximately 70%) or maternal uniparental disomy (UPD) (approximately 25%). Most cases of AS result from the lack of a maternal contribution from this same region, by maternal deletion (70%) or paternal UPD (approximately 5%). Analysis of allelic methylation differences at the small nuclear ribonucleoprotein polypeptide N (SNRPN) locus differentiates the maternally and paternally inherited chromosome 15 and can be used as a diagnostic test for AS and PWS.

Methods: Methylation-sensitive high-resolution melting-curve analysis (MS-HRM) using the DNA binding dye EvaGreen was used to analyze methylation differences at the SNRPN locus in anonymized DNA samples from individuals with PWS (n = 39) or AS (n = 31) and from healthy control individuals (n = 95). Results from the MS-HRM assay were compared to those obtained by use of a methylation-specific PCR (MSP) protocol that is used commonly in diagnostic practice.

Results: With the MS-HRM assay 97.6% of samples were unambiguously assigned to the 3 diagnostic categories (AS, PWS, normal) by use of automated calling with an 80% confidence percentage threshold, and the failure rate was 0.6%. One PWS sample showed a discordant result for the MS-HRM assay compared to MSP data.

Conclusions: MS-HRM is a simple, rapid, and robust method for screening methylation differences at the SNRPN locus and could be used as a diagnostic screen for PWS and AS.
Development and Validation of an Automated Thawing and Mixing Workcell
Authors: Hawker Charles D , Roberts William L , DaSilva Antonio , Stam Gordon D , Owen William E , Curtis DeVirl , Choi Byung-Sang , Ring Terry A
Published: September 2007

Background: Working toward a goal of total laboratory automation, we are automating manual activities in our highest volume laboratory section. Because half of all specimens arriving in this laboratory section are frozen, we began by developing an automated workcell for thawing frozen specimens and mixing the thawed specimens to remove concentration gradients resulting from freezing and thawing.

Methods: We developed an initial robotic workcell that removed specimens from the transport system’s conveyor, blew high-velocity room temperature air at the tubes, mixed them, and replaced them on the conveyor. Aliquots of citrated plasma were frozen with thermocouples immersed in the tubes, and thawing times and temperatures were monitored. Completeness of mixing of thawed specimens was studied by careful removal of small aliquots from the uppermost layer of the upright tubes without disturbing tube contents and analysis of total protein and electrolytes.

Results: High velocity ambient air aimed directly at tubes ranging from 12 × 75 to 16 × 100 mm brought specimens to room temperature in a maximum of 23 min. Adequate mixing of the specimens by the workcell’s robot required only 2 approximate 126° movements from an upright starting point, a surprising observation, because laboratorians are usually trained to mix 10 or 20 times. We also observed that, in a frozen overfilled tube, resulting analyte concentrations will be lower because more concentrated solutes leak from the tube.

Conclusions: A high-throughput, automated thawing and mixing workcell was successfully built, validated, and installed on our automated transport and sorting system.
Influence of Test Method on Direct Shear Behavior of Segmental Retaining Wall Units
Authors: Bathurst Richard J. , Althoff Sebastian , Linnenbaum Peter
Published: September 2007

This paper reports the results of interface shear testing carried out to investigate the influence of test methodology on shear-displacement behavior of five different segmental (modular) masonry concrete block configurations. The tests were carried out in general conformance with existing ASTM and National Concrete Masonry Association test protocols. Three different normal load arrangements were investigated: (1) flexible airbag, (2) fixed vertical piston, and (3) an adjustable vertical piston. A video-extensometer camera device was used to record block deformations in the vertical plane during testing. The test results showed that the three load arrangements gave similar shear capacity failure envelopes for the frictional block system with flat concrete surfaces. For more typical block systems with concrete shear keys and trailing lips that exhibit dilatant interface shear behavior, or a system with shear pins, the most consistent test results were developed using the flexible airbag arrangement. The results of this paper can be used to guide the selection of the loading arrangement for conventional laboratory modular block shear testing.
Inputs and Analyses: An End User’s Perspective of Heat-Air-Moisture Data
Authors: Decareau Christopher P. , Kan Liyen , Piñon Joseph P.
Published: September 2007

Hygrothermal simulation programs (HSPs) have the potential to help the way building enclosures are designed if end users are convinced the results are reliable. However, HSPs suffer from a lack of manufacturer-specific material data. HSP users often “create” materials by scouring multiple sources for hygrothermal properties. To the less familiar HSP user, significant error can be introduced into simulations by using properties that are calculated from variable sources. Through a series of case studies, this paper illustrates how material properties can make a difference in hygrothermal calculations and, ultimately, material specification.
Validation of Oxidative Stability of Factory Fill and Alternative Engine Oils Using the Iron Catalyzed Oxidation Test
Authors: Fitamen Eric , Tiquet Laurent , Woydt Mathias
Published: September 2007

Oxidation resistance is one of the limiting factors for long drains, bionotox, LowSAP or NoSAP engine oils. The French iron catalyzed oxidation test (ICOT) uses 60 ppm iron acetylacetonate as diluted catalyst under an aeration of 10 L∕h at 170°C. The ICOT ranks the oxidation stability of oils according to three properties: a. the viscosity increase (e.g., max. Δη40°C<100 % or Δη100°C<100 %), b. the increase to an individual TAN (e.g., max. 7.5 mgKOH∕g) and c. the oxidative evaporation losses of max. 10 % at Δη=100 % due to oxidation. This paper describes the ICOT method standardized as GFC Lu 36T 03 and the obtainable accuracy. Also, results are compiled achieved with this test for factory fill engine oils as well as alternative prototype engine oils based on esters, blends of esters with hydrocarbons, and polyglycols.
Moisture Buffering Capacities of Five North American Building Materials
Authors: Wu Yang , Fazio Paul , Kumaran Mavinkal K. (Kumar)
Published: September 2007

The moisture buffering capacities of five building materials which are widely used in North America was investigated experimentally. An experiment to quantify the moisture buffering capacity of building materials was developed. Hygrothermal properties of these building materials, reported in this paper, include the density, thermal conductivity, equilibrium moisture content, water vapor permeability, water absorption coefficient, and air permeability. Moisture buffering test results showed that among these materials the fiberboard product has the best moisture buffering capacity, whereas plywood has the lowest among the building products tested.
Expression of Cyp21a1 and Cyp11b1 in the fetal mouse testis
Authors: Hu Liangbiao , Monteiro Ana , Johnston Heather , King Peter , O’Shaughnessy Peter J
Published: September 2007

Fetal Leydig cells and fetal adrenocortical cells may share a common progenitor cell. Both cell types show several similarities, particularly in relation to their primary steroidogenic function. Differences in steroid secretion are largely due to the expression of 21-hydroxylase (CYP21A1) and 11β-hydroxylase (CYP11B1) activity in the adrenal. To determine whether expression of these enzymes defines a clear difference between adrenocortical and Leydig cells, or is further evidence of a link between the cell types, we have measured Cyp21a1 and Cyp11b1 expression and related enzyme activity in the fetal testis. Expression of both Cyp21a1 and Cyp11b1 was clearly detectable in the fetal testis by RT-PCR and Southern blotting. Real-time PCR studies showed that Cyp11b1 was expressed only in the fetal/neonatal testis with no expression in the pubertal or post-pubertal animal. Cyp21a1 was also predominantly expressed in the fetal testis although some lower expression was also seen in the adult. Expression of Cyp21a1 and Cyp11b1 in neonatal testicular cells was unaffected by incubation in vitro with human chorionic gonadotrophin or ACTH. Using immunohistochemistry, CYP21A1 was localised to a subset of interstitial steroidogenic cells in the fetal testis although CYP11B1 was not detectable. Incubation studies showed that 21-hydroxylase activity was present in the tissue although 11β-hydroxylase activity could not be detected. Results indicate that a subpopulation of steroidogenic cells in the fetal testis express Cyp21a1 and show 21-hydroxylase activity. This may provide further evidence of a link between fetal Leydig cells and adrenocortical cells but does not discount the possibility that these steroidogenic cells represent ‘ectopic’ adrenal cells.
KIT receptor-positive cells in the bovine corpus luteum are primarily theca-derived small luteal cells
Authors: Spanel-Borowski Katharina , Sass Kristina , Löffler Sabine , Brylla Elke , Sakurai Michiharu , Ricken Albert M
Published: September 2007

The tyrosine kinase KIT receptor, the protooncogene CD117, plays a key role in growth and maturation of oocytes and follicles. Relevant data are sparse for the corpus luteum (CL). We first confirmed the presence of KIT mRNA and KIT protein in bovine CL homogenates. We then localized KIT-positive (KIT+) cells in CL sections by immunohistochemistry. At the CL stage of early development, the former theca transforming into capsule/septa showed a strong band-like KIT+ immunoresponse. In addition, CD45+ leukocytes in septa included subpopulations of CD45+/KIT+ and CD14+/KIT+ leukocytes as validated by double immunofluorescence localization. At the early secretory stage, KIT+ cells appeared within the septa/capsule region and in the periphery of the CL parenchyma, there forming a complex network. This was separate from the capillary bed as determined by double staining for CD117 and FVIII-related endothelial cell antigen (FVIIIr). The KIT+ network coincided with cells positive for cytochrome P450 17α-hydroxylase, a thecal cell-specific enzyme. The late secretory stage was defined by an advanced manifestation of the KIT+ network in the CL periphery. At the stage of regression, the KIT+ network was absent. The CL of pregnancy expressed high levels of KIT mRNA and KIT protein uniformly throughout pregnancy. The KIT+ immunolocalization revealed small fibroblast-like cells, luteal cells with granules, and clusters of large luteal cells with staining of the cell membrane. We conclude that a majority of KIT+ cells in the bovine CL are primarily theca-derived small luteal cells, and that a minority represent KIT+ leukocytes, in some cases KIT+ monocytes.
Evaluation of the Storage Root‐Forming Legume Yam Bean (Pachyrhizus spp.) under West African Conditions
Authors: Zanklan A. Séraphin , Ahouangonou Salomon , Becker Heiko C. , Pawelzik Elke , Grüneberg Wolfgang J.
Published: September 2007

The yam bean (Pachyrhizus spp.) contains three closely related cultivated species: P. tuberosus (Lam.) Sprengel, P. erosus (L.) Urban, and P. ahipa (Wedd.). Its storage root dry matter content (SRDM) is usually low, although genotypes with a high SRDM have been identified (‘Chuin’ accessions). Flowers are often removed through flower pruning (FP) to increase storage root fresh matter yield (SRFY). The main objectives of this study were to investigate the potential for use in Benin (West Africa), to estimate the effect of FP, and to test whether roots could be processed into gari In total, 34 accessions were tested at one drought‐stress and one irrigated location. Means and genetic variance components were estimated for 33 agronomic traits. Without FP, the SRFYs of P. tuberosus, P. erosus, and P. ahipa were 13.9, 23.4, and 12.4 t ha−1, respectively, and the seed yields were 2.2, 5.2, and 2.1 t ha−1, respectively. The FP caused SRFY to increase by 48, 91, and 61% in P. tuberosus, P. erosus, and P. ahipa, respectively. The storage root dry matter yield (SRDY) of P. erosus was only slightly higher (∼8.5 t ha−1) than that of the Chuin accessions (∼8.0 t ha−1). Under drought, the SRDY was least affected in P. erosus Early‐maturing P. ahipa accessions were identified. All species could be used to make gari, which contained, on average, 5.5% protein, 58.5% starch, and 23.8% total dietary fiber. The crop has the potential for use in West Africa and has a large genetic variation for genetic improvement.
Photosynthetic Response of Cotton to Spider Mite Damage: Interaction with Light and Compensatory Mechanisms
Authors: Reddall A. A. , Wilson L. J. , Gregg P. C. , Sadras V. O.
Published: September 2007

We investigated the photosynthetic responses of cotton (Gossypium hirsutum L.) leaves to two‐spotted spider mite (Tetranychus urticae K.) damage. Light‐response curves of mite‐infested (+M) and uninfested (− M) leaves diverged as mite populations increased. At 17 adult female mites per leaf, photosynthetic rate of +M leaves at photosynthetic photon flux density of about 1600 μmol m−2 s−1 was halved from 31 μmol CO2 m−2 s−1 in −M to 16 μmol CO2 m−2 s−1 in +M but there was no effect on either respiration or apparent maximum quantum yield. This has important implications when comparing the response to mites of individual leaves versus canopies. In the field (i) photosynthesis declined with crop age, but the rate of decline was faster in mite‐infested leaves, and (ii) mite damage progressed downward in the canopy and from basal to distal leaf positions. We found no evidence of within‐leaf (i.e., basal vs. distal section) or within‐plant (top vs. mid or bottom leaf) increases in photosynthesis in compensation for mite damage, except for a minor enhancement of photosynthesis in bottom leaves of mite‐infested crops due to greater light penetration in canopies severely defoliated by mite damage.
Genetic Diversity of Canadian Soybean Cultivars and Exotic Germplasm Revealed by Simple Sequence Repeat Markers
Authors: Fu Yong‐Bi , Peterson Gregory W. , Morrison Malcolm J.
Published: September 2007

Genetic diversity assessment of improved crop germplasm can facilitate the expansion of the genetic base in a plant breeding program, but little effort has been made to assess the Canadian soybean [Glycine max (L.) Merr.] gene pool established over the past century. Simple sequence repeat (SSR) markers were applied to assess the genetic diversity of 45 Canadian soybean cultivars released from 1934 to 2001 and 37 exotic germplasm accessions. Thirty‐seven SSR primer pairs were applied and 234 polymorphic bands were scored for each accession. The frequencies of the scored bands ranged from 0.01 to 0.90 and averaged 0.17. The proportion of total SSR variation occurring between exotic and Canadian germplasm was 9%; among the Canadian cultivars of three breeding periods 10%; and between the cultivars of maturity groups 0 and 00 4%. More diversity was found for exotic germplasm than the Canadian. More diversity was observed in the cultivars of the recent breeding period than the early. The Canadian cultivars were clustered into seven major groups, partially congruent to the known pedigrees, and they were more related to germplasm from Russia, Sweden, and Ukraine and less to the Asian germplasm. The six genetically most distinct cultivars were PS86 RR, Gaillard, Manitoba Brown, Beechwood, Maple Isle, and 92B91. These findings are useful for the selection of genetically distinct or less related soybean materials to improve the genetic background of the soybean gene pool.
Nasal Airway Resistance at Different Climate Exposures: Description of a Climate Aggregate and Its Use
Authors: Ivarsson Alf , Malm Lars
Published: September 2007

A climate aggregate is described which can quickly change the flow and the relative humidity of air to be inhaled through the human nose, and, somewhat more slowly, can change the temperature. The flow can be changed from 0 to 3 liters/sec within seconds, and the relative humidity at +21°C from 90% to 10% within 3.5 minutes. Rhinomanometry was used to measure reactions in the nose to changed climates of the air to be inhaled. When the relative humidity of the air of +21°C was reduced from 90% to 10%, no change of the nasal airway resistance occurred.
Cellular Infiltration in Allergic Late-Phase Reaction of the Nose: Immunohistochemical Studies
Authors: Bachert Claus , Schindelbeck Gerhard , Hauser Ulrich
Published: September 2007

As with the bronchial system and the skin, the human nose may react with an immediate-phase (IPR) and late-phase reaction (LPR) upon allergen exposure. In this study we challenged intranasally 11 unselected grass-pollen-allergic patients and three normal controls with increasing allergen concentrations to provoke similar IPR symptoms in each person. A nasal symptom score and the bilateral nasal resistance were measured for 8 hours, repeated mucosal surface cell samples were performed (brush technique) before and every 2 hours after the challenge for histological and immunohistochemical investigations on cellular events during the nasal LPR. Apart from neutrophil and eosinophil infiltration, the LPR was accompanied by the migration of metachromatic cells, IgE positive cells, and macrophages/monocyte-like cells. IgE-positive cells could be identified as mast cells/basophils; from the discrepancy between the numbers of IgE- and metachromatic cell types we conclude the degranulation of these mediator cells not only during the IPR, but also during the LPR. The migration of IGE-bearing mediator cells onto the mucosal surface may represent an explanation for the increased reactivity of the organ due to repeated allergen exposure. Furthermore, the migration points to these cells as a means of transport for IgE molecules. The increase in density of macrophages/monocyte-like cells on the nasal mucosal surface during the LPR suggests an important role of these inflammatory cells in antigen processing and/or maintaining of chronic inflammatory reactions.
Blood Eosinophilia and Nasal Polyps
Authors: Wong Dennis , Jordana Gloria , Denburg Judah , Dolovich Jerry
Published: September 2007

In a controlled prospective study, patients who had nasal polypectomy were compared with patients who required other nasal surgery. Patients with and without asthma were identified. Both polyposis and asthma were independently associated with increased blood absolute eosinophil counts. Absolute eosinophil counts were: neither disease 0.199 × 106 cell/mL ± 0.025, polyposis alone 0.249 × 106 cell/mL ± 0.017, asthma alone 0.318 × 106 cell/mL ± 0.092, and both 0.375 × 106 cell/mL ± 0.076. In contrast, we were unable to show a difference between polyp patients and the controls in the serum IgE concentration or allergy skin test reactivity. Atopy was not a predictive factor for asthma in polyp patients. We conclude that nasal polyposis, like asthma, is highly associated with eosinophilia.
Trigeminal Traumatic Neuroma, A Previously Unreported Complication of Sinus Surgery: Nerve Exposure and Traumatic Neuroma of V2 Secondary to Maxillary Antrostomy
Authors: McGrew Robert N.
Published: September 2007

Sharp lancinating pain on inhalation as one type of atypical facial pain may be indicative of nerve exposure to the nasal airstream. A case is reported demonstrating iatrogenic nerve exposure of V2 which occurred as a complication of antrostomy. Development of a traumatic neuroma followed with gradual worsening of the pain. A method of management that was adequate in controlling the pain is described. Anatomic and surgical implications are discussed.