bluuebunny/crossref_metadata_2025 huggingface dataset:
The balance between RuBP carboxylation and RuBP regeneration: a mechanism underlying the interspecific variation in acclimation of photosynthesis to seasonal change in temperature
Authors: Onoda Yusuke , Hikosaka Kouki , Hirose Tadaki
Published: October 2005

The ratio of the capacities of ribulose-1,5-bisphosphate (RuBP) regeneration to RuBP carboxylation (Jmax / Vcmax) (measured at a common temperature) increases in some species when they are grown at lower temperatures, but does not increase in other species. To investigate the mechanism of interspecific difference in the response of Jmax / Vcmax to growth temperature, we analysed the temperature dependence of Vcmax and Jmax in Polygonum cuspidatum and Fagus crenata with the Arrhenius function. P. cuspidatum had a higher ratio of Jmax / Vcmax in spring and autumn than in summer, while F. crenata did not show such change. The two species had a similar activation energy for Vcmax (EaV) across seasons, but P. cuspidatum had a higher activation energy for Jmax (EaJ) than F. crenata. Reconstruction of the temperature response curve of photosynthesis showed that plants with an inherently higher EaJ / EaV (P. cuspidatum) had photosynthetic rates that were limited by RuBP regeneration at low temperatures and limited by RuBP carboxylation at high temperatures, while plants with an inherently lower EaJ / EaV (F. crenata) had photosynthetic rates that were limited solely by RuBP carboxylation over the range of temperatures. These results indicate that the increase in Jmax / Vcmax at low growth temperatures relieved the limitation of RuBP regeneration on the photosynthetic rate in P. cuspidatum, but that such change in Jmax / Vcmax would not improve the photosynthetic rate in F. crenata. We suggest that whether or not the Jmax / Vcmax ratio changes with growth temperature is attributable to interspecific differences in EaJ / EaV between species.
Detection of the placental epigenetic signature of themaspingene in maternal plasma
Authors: Chim Stephen S. C. , Tong Yu K. , Chiu Rossa W. K. , Lau Tze K. , Leung Tse N. , Chan Lisa Y. S. , Oudejans Cees B. M. , Ding Chunming , Lo Y. M. Dennis
Published: October 2005

The discovery of fetal DNA in the plasma of pregnant women has opened up new approaches for noninvasive prenatal diagnosis and monitoring. Up to now, the lack of a fetal DNA marker that can be universally detected in maternal plasma has limited the clinical application of this technology. We hypothesized that epigenetic differences between the placenta and maternal blood cells could be used for developing such a marker. By using bisulfite DNA sequencing, the methylation status of themaspingene promoter in placental tissues and paired maternal blood cells from pregnant women was analyzed. Themaspingene promoter was found to be hypomethylated in placental tissues and densely methylated in maternal blood cells. Genotyping of a single nucleotide polymorphism within the unmethylatedmaspinsequences in maternal plasma demonstrated that these sequences were derived from the fetus. By using real-time quantitative methylation-specific PCR, unmethylatedmaspinsequences were detected in maternal plasma in all three trimesters of pregnancy and were cleared within 24 h after delivery. The maternal plasma concentration of unmethylatedmaspinsequences was elevated by a median of 5.7 times in preeclamptic pregnancies compared with nonpreeclamptic pregnancies. HypomethylatedmaspinDNA is the first universal marker for fetal DNA in maternal plasma, thus allowing the measurement of fetal DNA concentrations in pregnancy-associated disorders, irrespective of fetal gender and genetic polymorphisms. Differential DNA methylation between the placenta and maternal blood cells may be exploited to develop further markers for noninvasive prenatal assessment.
Mechanism of radiation-induced bystander effect: Role of the cyclooxygenase-2 signaling pathway
Authors: Zhou Hongning , Ivanov Vladimir N. , Gillespie Joseph , Geard Charles R. , Amundson Sally A. , Brenner David J. , Yu Zengliang , Lieberman Howard B. , Hei Tom K.
Published: October 2005

The radiation-induced bystander effect is defined as “the induction of biological effects in cells that are not directly traversed by a charged particle but are in close proximity to cells that are.” Although these bystander effects have been demonstrated with a variety of biological endpoints in both human and rodent cell lines (as well as in 3D tissue samples), the mechanism of the phenomenon is not known. Although gap junction communication and the presence of soluble mediator(s) are both known to play important roles in the bystander response, the precise signaling molecules have yet to be identified. By using the Columbia University charged particle beam in conjunction with a strip dish design, we show here that the cyclooxygenase-2 (COX-2, also known as prostaglandin endoperoxide synthase-2) signaling cascade plays an essential role in the bystander process. Treatment of bystander cells with NS-398, which suppresses COX-2 activity, significantly reduced the bystander effect. Because the critical event of the COX-2 signaling is the activation of the mitogen-activated protein kinase pathways, our finding that inhibition of the extracellular signal-related kinase phosphorylation suppressed bystander response further confirmed the important role of mitogen-activated protein kinase signaling cascade in the bystander process. These results provide evidence that the COX-2-related pathway, which is essential in mediating cellular inflammatory response, is the critical signaling link for the bystander phenomenon.
The experimental folding landscape of monomeric lactose repressor, a large two-domain protein, involves two kinetic intermediates
Authors: Wilson Corey J. , Das Payel , Clementi Cecilia , Matthews Kathleen S. , Wittung-Stafshede Pernilla
Published: October 2005

To probe the experimental folding behavior of a large protein with complex topology, we created a monomeric variant of the lactose repressor protein (MLAc), a well characterized tetrameric protein that regulates transcription of the
lac
operon. Purified MLAc is folded, fully functional, and binds the inducer isopropyl β-
d
-thiogalactoside with the same affinity as wild-type LacI. Equilibrium unfolding of MLAc induced by the chemical denaturant urea is a reversible, apparent two-state process (pH 7.5, 20°C). However, time-resolved experiments demonstrate that unfolding is single-exponential, whereas refolding data indicate two transient intermediates. The data reveal the initial formation of a burst-phase (τ < ms) intermediate that corresponds to ≈50% of the total secondary-structure content. This step is followed by a rearrangement reaction that is rate-limited by an unfolding process (τ ≈ 3 s; pH 7.5, 20°C) and results in a second intermediate. This MLAc intermediate converts to the native structure (τ ≈ 30 s; pH 7.5, 20°C). Remarkably, the experimental folding-energy landscape for MLAc is in excellent agreement with theoretical predictions using a simple topology-based C
α
-model as presented in a companion article in this issue.
Multiplexed variation scanning for 1,000 amplicons in hundreds of patients using mismatch repair detection (MRD) on tag arrays
Authors: Faham Malek , Zheng Jianbiao , Moorhead Martin , Fakhrai-Rad Hossein , Namsaraev Eugeni , Wong Kee , Wang Zhiyong , Chow Shu G. , Lee Liana , Suyenaga Kent , Reichert Jennifer , Boudreau Andrew , Eberle James , Bruckner Carsten , Jain Maneesh , Karlin-Neumann George , Jones Hywel B. , Willis Thomas D. , Buxbaum Joseph D. , Davis Ronald W.
Published: October 2005

Identification of the genetic basis of common disease may require comprehensive sequence analysis of coding regions and regulatory elements in patients and controls to find genetic effects caused by rare or heterogeneous mutations. In this study, we demonstrate how mismatch repair detection on tag arrays can be applied in a case-control study. Mismatch repair detection allows >1,000 amplicons to be screened for variations in a single laboratory reaction. Variation scanning in 939 amplicons, mostly in coding regions within a linkage peak, was done for 372 patients and 404 controls. In total, >180 Mb of DNA was scanned. Several variants more prevalent in patients than in controls were identified. This study demonstrates an approach to the discovery of susceptibility genes for common disease: large-scale direct sequence comparison between patients and controls. We believe this approach can be scaled up to allow sequence comparison in the whole-genome coding regions among large sets of cases and controls at a reasonable cost in the near future.
A human monoclonal antibody neutralizes diverse HIV-1 isolates by binding a critical gp41 epitope
Authors: Miller Michael D. , Geleziunas Romas , Bianchi Elisabetta , Lennard Simon , Hrin Renee , Zhang Hangchun , Lu Meiqing , An Zhiqiang , Ingallinella Paolo , Finotto Marco , Mattu Marco , Finnefrock Adam C. , Bramhill David , Cook James , Eckert Debra M. , Hampton Richard , Patel Mayuri , Jarantow Stephen , Joyce Joseph , Ciliberto Gennaro , Cortese Riccardo , Lu Ping , Strohl William , Schleif William , McElhaugh Michael , Lane Steven , Lloyd Christopher , Lowe David , Osbourn Jane , Vaughan Tristan , Emini Emilio , Barbato Gaetano , Kim Peter S. , Hazuda Daria J. , Shiver John W. , Pessi Antonello
Published: October 2005

HIV-1 entry into cells is mediated by the envelope glycoprotein receptor-binding (gp120) and membrane fusion-promoting (gp41) subunits. The gp41 heptad repeat 1 (HR1) domain is the molecular target of the fusion-inhibitor drug enfuvirtide (T20). The HR1 sequence is highly conserved and therefore considered an attractive target for vaccine development, but it is unknown whether antibodies can access HR1. Herein, we use gp41-based peptides to select a human antibody, 5H/I1-BMV-D5 (D5), that binds to HR1 and inhibits the assembly of fusion intermediates
in vitro
. D5 inhibits the replication of diverse HIV-1 clinical isolates and therefore represents a previously unknown example of a crossneutralizing IgG selected by binding to designed antigens. NMR studies and functional analyses map the D5-binding site to a previously identified hydrophobic pocket situated in the HR1 groove. This hydrophobic pocket was proposed as a drug target and subsequently identified as a common binding site for peptide and peptidomimetic fusion inhibitors. The finding that the D5 fusion-inhibitory antibody shares the same binding site suggests that the hydrophobic pocket is a “hot spot” for fusion inhibition and an ideal target on which to focus a vaccine-elicited antibody response. Our data provide a structural framework for the design of new immunogens and therapeutic antibodies with crossneutralizing potential.
A nitric oxide processing defect of red blood cells created by hypoxia: Deficiency of S-nitrosohemoglobin in pulmonary hypertension
Authors: McMahon Timothy J. , Ahearn Gregory S. , Moya Martin P. , Gow Andrew J. , Huang Yuh-Chin T. , Luchsinger Benjamin P. , Nudelman Raphael , Yan Yun , Krichman Abigail D. , Bashore Thomas M. , Califf Robert M. , Singel David J. , Piantadosi Claude A. , Tapson Victor F. , Stamler Jonathan S.
Published: October 2005

The mechanism by which hypoxia [low partial pressure of O
2
(pO
2
)] elicits signaling to regulate pulmonary arterial pressure is incompletely understood. We considered the possibility that, in addition to its effects on smooth muscle, hypoxia may influence pulmonary vascular tone through an effect on RBCs. We report that exposure of native RBCs to sustained hypoxia is accompanied by a buildup of heme iron-nitrosyl (FeNO) species that are deficient in pO
2-
governed intramolecular transfer of NO to cysteine thiol, yielding a deficiency in the vasodilator
S
-nitrosohemoglobin (SNO-Hb).
S
-nitrosothiol (SNO)-deficient RBCs produce impaired vasodilator responses
in vitro
and exaggerated pulmonary vasoconstrictor responses
in vivo
and are defective in oxygenating the blood. RBCs from hypoxemic patients with elevated pulmonary arterial pressure (PAP) exhibit a similar FeNO/SNO imbalance and are thus deficient in pO
2
-coupled vasoregulation. Chemical restoration of SNO-Hb levels in both animals and patients restores the vasodilator activity of RBCs, and this activity is associated with improved oxygenation and lower PAPs.
How defects in central tolerance impinge on a deficiency in regulatory T cells
Authors: Chen Zhibin , Benoist Christophe , Mathis Diane
Published: October 2005

Both central (thymic) and peripheral (nonthymic) mechanisms are important for the induction and maintenance of T cell tolerance. Mice with a defect in Foxp3, required for the generation and activity of CD4+CD25+regulatory T cells, exhibit massive lymphoproliferation and severe inflammatory infiltration of multiple organs, in particular the lungs, liver, and skin. We have explored how this phenotype is influenced by an additional defect in central tolerance induction, generated by either crossing in a null mutation of theAiregene or substituting the nonobese diabetic (NOD) genetic background. The double-deficient mice had fulminant autoimmunity in very early life and a gravely shortened lifespan vis-à-vis single-deficient littermates. They showed massive lymphoproliferation and exacerbated inflammatory damage, particularly in the lungs and liver. Yet, the range of affected sites was not noticeably extended, and, surprisingly, many organs, or regions of organs, remained untouched, suggesting additional important mechanisms to enforce immunological self-tolerance.
Identification of K-ras as the major regulator for cytokine-dependent Akt activation in erythroid progenitors
in vivo
Authors: Zhang Jing , Lodish Harvey F.
Published: October 2005

Despite intensive investigation, controversial results have been obtained concerning the precise signaling pathway(s) regulated by K-ras in different cell types. We show that in primary fetal liver erythroid progenitors, erythropoietin activates all three Ras isoforms, but preferentially N- and K-ras. In K-
ras
-/-
fetal liver cells (FLC), erythropoietin- or stem cell factor-dependent Akt activation is greatly reduced, whereas other pathways including Stat5 and p44/p42 MAP kinase are activated normally. We further studied the effects of reduced cytokine-dependent Akt activation in erythroid differentiation. We find that freshly isolated K-
ras
-/-
FLC show an ≈7-fold increase of apoptosis and delayed erythroid differentiation, but only at the stage of erythroid progenitors and very early erythroblasts. When K-
ras
-/-
erythroid progenitors are cultured
in vitro
, there is a significant delay in erythroid differentiation but little increase in apoptosis. Furthermore, we show that partial pharmacologic inhibition of the phosphatidylinositol 3-kinase/Akt pathway in wild-type erythroid progenitors leads to a delay in erythroid differentiation similar to that observed in K-
ras
-/-
FLC. Taken together, our data identify K-ras as the major regulator for cytokine-dependent Akt activation, which is important for erythroid differentiation
in vivo
.
γδ T cell homeostasis is established in competition with αβ T cells and NK cells
Authors: French Jena D. , Roark Christina L. , Born Willi K. , O'Brien Rebecca L.
Published: October 2005

γδ T cells are a diverse population of lymphocytes that play an important role in immune regulation. The size of the γδ T cell pool is tightly regulated, comprising only 1-10% of total lymphoid T cells in mice and humans. We examined the homeostatic regulation of γδ T cells using a model of lymphopenia-induced homeostatic expansion. We found that IL-15 and, to a lesser extent, IL-7 play an important role in lymphoid γδ T cell homeostasis. Moreover, γδ T cell homeostatic expansion was limited not only by γδ T cells themselves but also by natural killer cells and αβ T cells. Our results suggest that CD8+αβ T cells are the most potent inhibitors of γδ T cell homeostasis and exert their effect by competing for IL-15.
The extracellular region of ErbB4 adopts a tethered conformation in the absence of ligand
Authors: Bouyain Samuel , Longo Patti A. , Li Shiqing , Ferguson Kathryn M. , Leahy Daniel J.
Published: October 2005

The human ErbB family of receptor tyrosine kinases comprises the epidermal growth factor receptor (EGFR/ErbB1/HER1), ErbB2 (HER2/Neu), ErbB3 (HER3), and ErbB4 (HER4). ErbBs play fundamental roles in cell growth and differentiation events in embryonic and adult tissues, and inappropriate ErbB activity has been implicated in several human cancers. We report here the 2.4 Å crystal structure of the extracellular region of human ErbB4 in the absence of ligand and show that it adopts a tethered conformation similar to inactive forms of ErbB1 and ErbB3. This structure completes the gallery of unliganded ErbB receptors and demonstrates that all human ligand-binding ErbBs adopt the autoinhibited conformation. We also show that the binding of neuregulin-1β to ErbB4 and ErbB3 and the binding of betacellulin to both ErbB4 and ErbB1 does not decrease at low pH, unlike the binding of epidermal growth factor and transforming growth factor-α to ErbB1. These results indicate an important role for ligand in determining pH-dependent binding and may explain different responses observed when the same ErbB receptor is stimulated by different ligands.
Cholesterol-induced macrophage apoptosis requires ER stress pathways and engagement of the type A scavenger receptor
Authors: DeVries-Seimon Tracie , Li Yankun , Yao Pin Mei , Stone Elizabeth , Wang Yibin , Davis Roger J. , Flavell Richard , Tabas Ira
Published: October 2005

Macrophage death in advanced atherosclerosis promotes necrosis and plaque destabilization. A likely cause of macrophage death is accumulation of free cholesterol (FC) in the ER, leading to activation of the unfolded protein response (UPR) and C/EBP homologous protein (CHOP)–induced apoptosis. Here we show that p38 MAPK signaling is necessary for CHOP induction and apoptosis. Additionally, two other signaling pathways must cooperate with p38-CHOP to effect apoptosis. One involves the type A scavenger receptor (SRA). As evidence, FC loading by non-SRA mechanisms activates p38 and CHOP, but not apoptosis unless the SRA is engaged. The other pathway involves c-Jun NH2-terminal kinase (JNK)2, which is activated by cholesterol trafficking to the ER, but is independent of CHOP. Thus, FC-induced apoptosis requires cholesterol trafficking to the ER, which triggers p38-CHOP and JNK2, and engagement of the SRA. These findings have important implications for understanding how the UPR, MAPKs, and the SRA might conspire to cause macrophage death, lesional necrosis, and plaque destabilization in advanced atherosclerotic lesions.
Mozart's Archaic Endings: A Linguistic Critique
Authors: Rumph Stephen
Published: October 2005

Rhetorical studies of Mozart have assumed a rationalist conception of language, ignoring the empiricist model that actually dominated the Enlightenment. The two models, comparable structurally to thestile anticoandstyle galant, collide in Mozart's learned finales. A study of three finales, from the Mass in C minor, the Concerto in E♭, K.449, andDie Entführung aus dem Serail, shows how Mozart negotiated irreducible contradictions within Enlightenment thought by switching between the two models.
Fecal Tumor Necrosis Factor‐α and Calprotectin as Differential Diagnostic Markers for Severe Diarrhea of Small Infants
Authors: Kapel N , Roman C , Caldari D , Sieprath F , Canioni D , Khalfoun Y , Goulet O , Ruemmele F M
Published: October 2005

Objective:

The etiology of early‐onset diarrhea of neonates and small infants that persists despite bowel rest is heterogeneous. Two different categories of disorders presenting with diarrhea in the first weeks of life can be distinguished: constitutive intestinal epithelial disorders (microvillus atrophy [MVA] or epithelial dysplasia [ED]) and immune‐inflammatory disorders, (autoimmune enteropathy [AIE] or inflammatory colitis [IC]). We aimed to evaluate in a prospective manner the use of fecal inflammatory markers in the differential diagnosis of severe persistent diarrhea.

Material and Patients:

Twenty‐five patients (17 males) were enrolled in this study (median age 8 months). Fourteen children had a constitutive enterocyte disorder (group 1: MVA = 8, ED = 6), and 11 patients had an immuno‐inflammatory disease (group 2: AIE = 5, IC = 6). Stool samples were collected at the time of diagnosis and stored at −80° until tumor necrosis factor (TNF)‐α and calprotectin were measured by enzyme‐linked immunoadsorbent assay.

Results:

No significant differences in age at onset of diarrhea or in stool volumes were observed between both groups. In group 1, fecal TNF‐α was undetectable/normal in 14 of 14 children, whereas group 2 showed dramatically elevated TNF‐α levels (mean 3,104, range 237‐18,078 pg/g) in 8 of 11 patients. Similarly, calprotectin levels were undetectable/normal in 14 of 14 patients in group 1 and highly raised in 11 of 11 patients in group 2 (median 1,145, range 375‐3,095 μg/g), P < 0.01. Under therapy, these inflammatory parameters normalized.

Conclusions:

Determination of fecal inflammatory markers is a simple method helping to distinguish constitutive from immuno‐inflammatory etiologies of severe persistent diarrhea. These data also suggest that constitutive enterocyte disorders are not accompanied by an inflammatory mucosal reaction.
Short‐Term Effect of Complementary Feeding Frequency on Total Ad Libitum Consumption in 6‐ to 10‐Month‐Old Breast Fed Indian Infants
Authors: Singh P K , Sachdev H P. S , Nagpal J , Bajaj M , Dubey A P
Published: October 2005

Objective:

To evaluate the short‐term effect of frequency of complementary feeding on total ad libitum consumption in breast fed infants.

Methods:

Twenty infants between 6 to 10 months of age were studied in a tertiary hospital in New Delhi for 48 hours. A traditional gruel made of rice and pulses (mean (SD) caloric density 54.22 (7.08) kcal/100 g) was offered in a randomized manner three (n = 10) or four (n = 10) times per day to the subjects over the first 24 hours with the subjects crossing over in the next 24 hours. They were allowed ad libitum breast feeding with no other food or fluid during the study period. Total caloric intake from breast milk and semisolids was computed for each day.

Results:

There was no difference in the total caloric consumption with a semisolid feeding frequency of three or four times per day. The frequency of breast feeding and the breast feeding duration were also comparable (P > 0.05). However, breast milk intake was lower with a semisolid feeding frequency of 4 times/day (mean difference −61.2 g/d [95% confidence interval (CI) −122.2‐0.32]; P = 0.051). The time required for feeding was higher (mean difference 14.75 min; P < 0.001), whereas the per meal intake of semisolids was lower with four semisolid feeds per day (mean difference −5.5 kcal/meal; [95% CI −10.19 to −0.81]; P = 0.024).

Conclusion:

In the short term, a change in semisolid feeding frequency from three to four times per day does not result in enhanced energy consumption because of lower breast milk intake.
Language rights in Indigenous communities:  The case of the Inuit of Arctic Québec1
Authors: Patrick Donna
Published: October 2005

Despite the rise of language rights, it is not clear whether the granting of rights to Indigenous and minority groups has any direct effect on the ability of these groups to revitalize and maintain their languages. This paper offers an analysis of macro‐level rights discourse in Canada, drawing on certain Supreme Court of Canada decisions regarding Francophone‐minority and Aboriginal peoples. It then traces certain consequences of the granting of Indigenous language rights for Indigenous language instruction and maintenance, focusing on the Inuit in the Arctic Québec region of Nunavik.
Linguistic human rights as a source of policy guidelines: A critical assessment
Authors: Grin François
Published: October 2005

The ‘linguistic human rights’(LHR) perspective, though relevant to the protection and promotion of minority languages (or, more generally, of linguistic diversity), displays one major weakness, namely, its almost exclusive reliance on moral considerations. Although LHR advocates often back up their claims with other (including economic) arguments, the latter often fail to pass ‘hard’ validation tests. Hence, the actual import of LHR arguments is constrained by the extent to which the underpinning moral considerations are shared by public opinion and politicians. Even if there is broad consensus around values, arguments in favour of minority language rights can be defeated by three types of objections that have to do with feasibility, costs, and burden sharing. In order to overcome these objections, it is necessary to draw on the tools of policy evaluation, which provides strong arguments in favour of linguistic diversity.
Childhood acute lymphoblastic leukaemia and relapse
Authors: Gaynon Paul S.
Published: October 2005

Acute lymphoblastic leukaemia (ALL) is the most common childhood cancer. Treatment has improved but relapsed ALL remains more common than new cases of many ‘common’ paediatric malignancies. We have salvage regimens with substantial complete remission (CR) rates and increasing access to haematopoietic stem cell transplantation, but most patients who relapse die. We need better therapies. Insights into pharmacology may guide more effective use of existing agents. Novel agents with activity against resistant lymphoblasts offer an appealing strategy. However, most candidate agents fail, despite enthusiastic investigators, intriguing mechanisms of action and ‘compelling’ preclinical data. A number of existing combinations provide a 40% complete response rate in second or third relapse. Yet survival in third remission is <10%. Novel agents must, most likely, be integrated into multiagent combinations that provide a higher CR rate or better quality CR's than our conventional combinations in order to contribute substantially to cure. The march from bench to bedside requires careful consideration of the intermediate steps.
Comparison of apoptosis and mortality measurements in peripheral blood mononuclear cells (PBMCs) using multiple methods
Authors: Glisic‐Milosavljevic S. , Waukau J. , Jana S. , Jailwala P. , Rovensky J. , Ghosh S.
Published: October 2005

Abstract. Death through apoptosis is the main process by which aged cells that have lost their function are eliminated. Apoptotic cells are usually detected microscopically by changes in their morphology. However, determination of early apoptotic events is important for in vitro (and ex vivo) studies. The main objective of the present study is to find the most sensitive method for apoptosis detection in human peripheral blood mononuclear cells (PBMCs) by comparing six different methods following five different means of immunological stimulation at 3 and 5 days. Each of six apoptosis quantification methods, except the trypan blue exclusion test, is a combination of two stains, one for the specific detection of apoptotic cells and the other for the unspecific detection of dead cells. Values for apoptosis and mortality were compared with a reference method. The choice of apoptosis detection method is more important following 3 days of stimulation than after 5 days of stimulation (P = 2 × 10−6 versus P = 1 × 10−2). In contrast, we find mortality measurements following the different means of stimulation highly significant at both 3 and 5 days (F2.28 = 7.9, P = 1.4 × 10−6 at 3 days and F2.28 = 8.5, P = 4.5 × 10−7 at 5 days). Variation as a result of the combination of specific PBMC stimulation and the method used to detect apoptosis is reduced considerably with time (F1.58 + 3.7, P + 3 × 10−7 at 3 days to F = (1.58) = 0.97, P = 0.5 at 5 days). Based on Tukey's test, YO‐PRO‐1 is the most sensitive stain for apoptosis and, when combined with 7‐AAD, provides an accurate measure of apoptosis and mortality. In conclusion, we propose YO‐PRO‐1/7‐AAD as a new combination and low‐cost alternative for the sensitive detection of early apoptosis.
Macroprolactin does not contribute to elevated levels of prolactin in patients on renal replacement therapy
Authors: Yavuz Dilek , Topçu Güler , Özener Çetin , Akalın Sema , Sirikçi Önder
Published: October 2005

Objective  Three molecular forms of PRL with molecular weights of 23, 50–60 and > 100 kDa have been defined. The high‐molecular‐weight forms are called macroprolactin. Different immunoassays produce varyingly elevated results with macroprolactin‐containing sera. The kidneys are reported to clear 25% of PRL from the circulation. Hyperprolactinaemia is seen in 20–75% of patients with chronic renal failure (CRF). PRL clearance rate has been reported to be reduced in CRF and the resulting hyperprolactinaemia is due to reduced renal function.

Patients  To determine the contribution of macroprolactinaemia to elevated PRL levels in CRF, 91 patients receiving haemodialysis (HD), continuous ambulatory peritoneal dialysis (CAPD) and renal transplantation (RT) therapies and 72 control subjects were included in the study.

Measurements  Serum PRL levels were measured by a sandwich immunoassay with electrochemical detection. Following polyethylene glycol (PEG) precipitation, recovery ratios were calculated and samples with a recovery of < 50% were identified as having macroprolactin isoforms.

Results  The serum and supernatant PRL levels of CRF patients were significantly higher than those of the control group (P < 0·001). The serum PRL levels of HD and CAPD patients were significantly higher than those of the RT patients (P < 0·001). The serum PRL levels of the RT patients and the control group did not differ significantly (P > 0·05). A moderate correlation was found between PRL and creatinine levels (r = 0·609, P < 0·001).

Conclusions  The hyperprolactinaemia seen in renal replacement therapy is not associated with the presence of macroprolactin isoforms but with the decline in renal function.
The characteristics of beef patties containing different levels of fat and oat flour
Authors: Serdaroglu Meltem
Published: October 2005

Three concentrations of oat flour (OF), 0, 2 and 4% (w/w), were added to beef patties which had either 5, 10 or 20% fat levels. Raw and cooked patties were analysed for protein, fat, moisture, ash and pH. Patties were further evaluated for cooking characteristics, colour parameters (L*, a*, b*) and sensory properties. Moisture content decreased in raw patties as a result of the addition of OF but OF increased the moisture content of cooked patties. OF addition did not change the protein, fat and ash contents of either raw or cooked patties. Increasing fat concentration decreased cooking yield. OF improved the cooking characteristics of patties. Reducing the fat significantly decreased the lightness values of patties. Patties with OF had slightly higher b* values than control treatments. Increasing fat level increased juiciness, flavour and texture scores and 4% OF increased the juiciness scores, OF had no effect on other sensory properties.
The comparative evaluation of the nutrient composition and sensory attributes of four vegetables cooked by different methods
Authors: Kala Arunachalam , Prakash Jamuna
Published: October 2005

The following four vegetables; beans (Phaseolus vulgaris var. vulgaris), brinjal (Solanum melongena var. esculentum), knol‐khol (Brassica oleracea var. caulorapa) and radish (Raphanus raphanistrum satvius) were processed either by conventional, pressure or microwave cooking methods and were analysed for their nutritional and sensory profile. Raw samples were used as controls. Microwave cooking resulted in greater moisture loss when compared with vegetables cooked by other methods. The protein, fat, total ash, calcium, phosphorus, iron, and dietary fibre contents of the vegetables did not differ significantly in raw or cooked vegetables. Ascorbic acid was destroyed to the greatest extent by pressure cooking followed by microwave and conventional method. The sensory attributes of cooked vegetables, as evaluated by a paired preference test, revealed that there were no significant differences between cooked radish and knol‐khol but, in the case of beans and brinjal, microwave and conventionally cooked samples were preferred over pressure cooked ones.
The effect of supplementing goats milk with whey protein concentrate on textural properties of set‐type yoghurt
Authors: Herrero Ana M. , Requena Teresa
Published: October 2005

Yoghurt was manufactured from goat's milk and supplemented with 30 g L−1 of whey protein concentrate (WPC). The textural properties of the yoghurt were evaluated during the shelf‐life of the product and the textural characteristics of yoghurt made from cow's milk were used as a reference. The instrumental analyses used were the puncture test, stress relaxation test and texture profile analysis. The addition of WPC to goat's milk enhanced the textural characteristics of yoghurt. These advantageous attributes included increased firmness, hardness and adhesiveness. These attributes were quantitatively similar (P > 0.05) to those obtained from yoghurt made from cow's milk. In addition, the textural properties were maintained constant throughout the shelf‐life of the product.
Skeleton Photoperiods Alter Delayed‐Type Hypersensitivity Responses and Reproductive Function of Siberian Hamsters (Phodopus sungorus)
Authors: Gatien M. L. , Hotchkiss A. K. , Dhabhar F. S. , Nelson R. J.
Published: October 2005

Photoperiod (day length) can modulate immune function. Whether these photoperiodic effects on immune function are mediated directly by a circadian photoperiodic time measurement system or indirectly by nonspecific (e.g. stressful) effects of light is unknown. To discriminate between these two possibilities, Siberian hamsters (Phodopus sungorus) were housed in either long or short photoperiods (LD 16 : 8 h or LD 8 : 16 h) or in ‘skeleton’ long or short photoperiods (LD 1 : 14 h: LD 1 : 8 h or LD 1 : 6 h: LD 1 : 16 h). In the skeleton photoperiods, both long‐ and short‐day animals received 2 h of light per day. After 10 weeks in their respective photoperiods, hamsters were tested for an antigen specific immune response using a delayed type hypersensitivity (DTH) model. Reproductive and endocrine responses of hamsters in each of the skeleton photoperiods were equivalent to those in standard long or short days, respectively. Hamsters in skeleton short days and LD 8 : 16 increased DTH responses compared to hamsters in both long‐day groups. DTH responses were equivalent in both long‐day groups. These results suggest that the influences of day length on immune function potentially are due to circadian photoperiodic time measurement systems.
Impact of insecticide‐treated bed nets implementation on the genetic structure of Anopheles arabiensis in an area of irrigated rice fields in the Sahelian region of Cameroon
Authors: WONDJI C. , SIMARD F. , LEHMANN T. , FONDJO E. , SAMÈ‐EKOBO A. , FONTENILLE D.
Published: October 2005

Variation at 12 microsatellite loci was investigated to assess the impact of the implementation of insecticide‐treated bed nets (ITNs) on the genetic structure of Anopheles arabiensis in Simatou, a village surrounded by irrigated rice fields in the Sahelian area of Cameroon. The An. arabiensis population of Simatou was sampled twice before ITN implementation, and twice after. Effective population size estimates (Ne) were similar across each time point, except for the period closely following ITN introduction where a nonsignificant reduction was recorded. Hence, we believe that ITN implementation resulted in a temporary bottleneck, rapidly followed by a demographic expansion. The genetic diversity of the population was not significantly affected since different genetic parameters (allele number, observed and expected heterozygosities) remained stable. Low estimates of genetic differentiation between the populations from Simatou and Lagdo, separated by 300 km, suggested extensive gene flow among populations of An. arabiensis in the Sahelian region of Cameroon. A decrease in the susceptibility to deltamethrin was observed following ITN introduction, but no kdr mutation was detected and a metabolic resistance mechanism is probably involved. The temporary effect of ITNs on the genetic structure of An. arabiensis population suggests that, to optimize the success of any control programme of this species based on ITNs, the control area should be very large and the programme should be implemented for a long period of time.
Vasovagal reactions in blood donors during or immediately after blood donation
Authors: Zervou E. K. , Ziciadis K. , Karabini F. , Xanthi E. , Chrisostomou E. , Tzolou A.
Published: October 2005

summary.  The aim of the study was to estimate the type, incidence and causes of donor adverse reactions during and after blood donation in a Greek Blood Bank, where medical staff is responsible for donor selection. 12 173 blood donors were studied for adverse reactions. One‐hundred and seven (0·87%) donors had a vasovagal reaction during or after blood donation. Donors who gave blood occasionally had a significant greater incidence of reactions compared with volunteer donors (1·15 versus 0·53%) (P < 0·001). There was no significant difference between men and women (0·85 versus 0·95%). First‐time donors (1·7 versus 0·68%) and those under 30 years (1·15 versus 0·71%] had a significant greater possibility to have a reaction (P < 0·001). Twenty‐two of 107 (20·5%) donors had a syncopal reaction. There was not a causative correlation of haematocrit, haemoglobin, systolic and diastolic blood pressure, pulse rate and weight in women (except weight in men) in developing a reaction. The stressing experience of phlebotomy was the reason for the higher frequency of a reaction.

The incidence of reactions in our donors is lower than in other studies, and the possible reason for this is that only physicians are responsible for the selection of donors and trained personnel are careful of them during the donation process.
The age of stored red blood cell concentrates at the time of transfusion
Authors: Raat N. J. H. , Berends F. , Verhoeven A. J. , De Korte D. , Ince C.
Published: October 2005

summary.  In the past decade, studies suggesting a reduced oxygen delivery by stored red blood cell concentrates (RBCCs) have initiated a discussion about the use of fresh versus old blood. We determined whether old RBCCs represent a significant part of the total of RBCCs issued. The age of RBCCs at the time of transfusion was determined in 74 084 units during a 5‐year period in the Academic Medical Center, a main Dutch University Hospital. The mean (±SD) storage time of the total number of transfused RBCC was 19·4 ± 7·0 days, and 37% were older than 3 weeks.

As more than one‐third of the transfused RBCC units are stored for longer than 3 weeks, the research examining differences in oxygen delivery between fresh and stored RBCC is relevant for packed RBC transfusion practice.
Recovery from post‐operative anaemia
Authors: Wallis J. P. , Wells A. W. , Whitehead S. , Brewster N.
Published: October 2005

summary.  Acceptance of lower transfusion thresholds and shorter post‐operative stays results in patients leaving hospital after surgery with lower haemoglobin (Hb) than previously. We undertook a prospective observational study to assess the haematological response to post‐operative anaemia and to determine the utility of quality of life (QoL) measures in assessing the impact of anaemia on such patients. Thirty patients undergoing unilateral hip arthroplasty had blood samples taken and QoL questionnaires administered pre‐operatively and at 7, 28 and 56 days post‐operatively. Increased erythropoiesis was evident at day 7 post‐operatively. Approximately two‐thirds of the post‐operative Hb deficit was corrected by day 28. There was evidence of functional iron deficiency in more than one‐quarter of patients at day 56. QoL scores used did not show any relationship with Hb in the post‐operative period. Red cell 2,3‐diphosphoglycerate (2,3DPG) levels increased in proportion to the degree of post‐operative anaemia. We concluded that substantial recovery of Hb occurs between day 7 and day 28 post‐operatively. Complete recovery of Hb may be delayed beyond day 56 due to development of iron deficiency. Patients are at significant risk of developing post‐operative iron deficiency depending on operative blood loss and pre‐operative iron stores. Increased red cell 2,3DPG may offset the effect of anaemia on oxygen delivery. We found no evidence that anaemia produces a measurable effect on chosen QoL scores in the post‐operative period.
Cost‐effectiveness of pathogen inactivation for platelet transfusions in the Netherlands
Authors: Postma M. J. , Van Hulst M. , De Wolf J. T. M. , Botteman M. , Staginnus U.
Published: October 2005

summary. The objective of this study is to estimate cost‐effectiveness of pathogen inactivation for platelet transfusions in the Netherlands. We used decision tree analysis to evaluate the cost‐effectiveness of the addition of pathogen inactivation of pooled platelets to standard procedures for platelet transfusion safety (such as, donor recruitment and screening). Data on transfusions were derived from the University Medical Centre Groningen (the Netherlands) for 1997. Characteristics of platelet recipients (patient group, age, gender and survival) and data/assumptions on viral and bacterial risks were linked to direct and indirect costs/benefits of pathogen inactivation. Post‐transfusion survival was simulated with a Markov model. Standard methods for cost‐effectiveness were used. Cost‐effectiveness was expressed in net costs per life‐year gained (LYG) and estimated in baseline‐ and sensitivity analysis. Sensitivity was analysed with respect to various assumptions including sepsis risk, reduction of the discard rate and discounting. Stochastic analysis to derive 90% simulation intervals (SIs) was performed on sepsis risk. Net costs per LYG for pathogen inactivation were estimated €554 000 in the baseline‐weighted average over the three patient groups (90% SI: €354 000–1092 500). Sensitivity analysis revealed that cost‐effectiveness was insensitive to viral risks and indirect costing, but highly sensitive to the assumed excess transfusions required and discounting of LYG. Given relatively high net costs per LYG that are internationally accepted for blood transfusion safety interventions, our estimated cost‐effectiveness figures for pathogen inactivation may reflect acceptable cost‐effectiveness in this specific area. Two main assumptions of our model were that the pathogen inactivation was 100% effective in preventing transmission of the pathogens considered and was not associated with major and/or costly adverse reactions. Validation of several crucial parameters is required, in particular the Dutch risk for acquiring and dying of transfusion‐related sepsis.
Hepatitis B virus DNA in blood donors with anti‐HBc as a possible indicator of active hepatitis B virus infection in Yucatan, Mexico
Authors: García‐Montalvo B. M. , Farfán‐Ale J. A. , Acosta‐Viana K. Y. , Puerto‐Manzano F. I.
Published: October 2005

summary.  Hepatitis B virus (HBV) may be present in serum even when negative for HBV surface antigen (HBsAg). If routine screening of sera for anti‐HBV core antigen (anti‐HBc) is not done, low‐level HBV viraemia may not be identified. A study was done on the presence of HBV DNA in serum samples from Mexican blood donors negative for HBsAg. Sera from 158 volunteer blood donors, negative for HBsAg and anti‐HBs, but positive for anti‐HBc, were analysed using nested polymerase chain reaction (PCR). HBV DNA was detected in sera from 13 (8·23%) of the 158. Specificity of the PCR‐amplified products was corroborated using Southern blot. Single strand conformation polymorphism (SSCP) analysis showed identical SSCP‐banding patterns for all 13 PCR products, suggesting similar cDNA sequences. Occult HBV infection was observed in approximately 8% of anti‐HBc only donors. The absence of HBsAg in the blood of apparently healthy individuals may not be sufficient to ensure lack of circulating HBV, and blood containing anti‐HBc only may be infectious until proven otherwise.
Remission induction in a Jehovah's Witness patient with acute myeloid leukaemia using gemtuzumab ozogamicin
Authors: Bareford D. , Odeh B. , Narayanan S. , Wiltshire S.
Published: October 2005

summary.  A 40‐year‐old patient, who was a Jehovah's Witness, with acute myeloid leukaemia entered remission using a chemotherapeutic based regime aided by the addition of gemtuzumab ozogamicin without requiring any blood product support. The use of gemtuzumab ozogamicin may have helped avoid fatal pancytopenia. The use of gemtuzumab ozogamicin might be considered in similar situations.
Perceived safety of donor blood and blood substitutes for transfusion: the role of informational frame, patient groups and stress appraisals
Authors: Ferguson E. , Leaviss J. , Townsend E. , Fleming P. , Lowe K. C.
Published: October 2005

summary.  Patients express concerns about the safety of donated blood. Do they have similar concerns about potential ‘blood substitutes’ and does the way information is presented on these options alter patients' perceptions? A two (informational frame: gain or loss) by four (transfusion type: human donor blood, human haemoglobin, bovine haemoglobin or perfluorocarbon emulsion substitutes) by three (patient group: adult haemophiliac/leukaemia patients, relatives/friends of haemophiliac/leukaemia patients and controls) between‐subjects design was used. There were 82 patients, 118 relatives/friends and 263 controls from the UK. Blood substitutes were perceived as being significantly less safe than donor blood. Perceptions of safety were greater when transfusion information (regardless of transfusion type or patient group) was presented as gains rather than losses. Different demographic and psychological factors predicted perceived safety (e.g. sex) and perceived risk (e.g. age and experience).
Myeloablative allogeneic hematopoietic stem cell transplantation in elderly patients
Authors: Ditschkowski M , Elmaagacli AH , Trenschel R , Steckel NK , Koldehoff M , Beelen DW
Published: October 2005

Abstract:  This study aimed to evaluate the outcome following myeloablative allogeneic hematopoietic stem cell transplantation (SCT) among patients older than 50 yr of age. A total of 215 patients with a median age of 57 yr underwent allogeneic hematopoietic SCT for early (41%) or advanced (59%) hematologic malignancies. After a median follow‐up of 36 months a 10‐yr survival estimate of 56 ± 6% could be assessed for patients in early disease stages while patients with advanced diseases showed a significantly decreased survival probability of 31 ± 5% (p < 0.0002). Transplant related mortality (TRM) at day 100 and 365 post‐transplant was 13% and 30% for early but increased to 21% and 49% for advanced disease stages. As major determinants of TRM advanced disease stage (p < 0.0001) and occurrence of grades II–IV graft‐vs.‐host disease (GVHD) (p < 0.0001) were identified. These results show that hematopoietic SCT following myeloablative conditioning is also applicable to elderly patients whereas disease stage and high‐grade GVHD represent the essential prognostic factors for outcome.