Search for Event Rate Modulation in XENON100 Electronic Recoil Data.

We have searched for periodic variations of the electronic recoil event rate in the (2-6) keV energy range recorded between February 2011 and March 2012 with the XENON100 detector, adding up to 224.6 live days in total. Following a detailed study to establish the stability of the detector and its background contributions during this run, we performed an unbinned profile likelihood analysis to identify any periodicity up to 500 days. We find a global significance of less than 1σ for all periods, suggesting no statistically significant modulation in the data. While the local significance for an annual modulation is 2.8σ, the analysis of a multiple-scatter control sample and the phase of the modulation disfavor a dark matter interpretation. The DAMA/LIBRA annual modulation interpreted as a dark matter signature with axial-vector coupling of weakly interacting massive particles to electrons is excluded at 4.8σ.

[Euthanasia of patients in coma vigil. Results on German medical staff attitudes]. / Aktive Sterbehilfe bei Menschen im Wachkoma? Ergebnisse einer Einstellungsuntersuchung bei Arzten und Pflegenden.

An examination was made concerning doctors' and nursing staff's attitudes towards active euthanasia of patients suffering from coma vigil (2652 doctors and 5785 nursing staff were interviewed). This investigation made clear that most of the persons asked about this group of patients voted for a change in German laws following the Dutch example. There were noticeable differences observed between the professional groups. A majority (64.79%) were convinced that under certain circumstances it is justified to end intentionally the life of persons in a coma vigil. Of the nursing staff, 70.38% were in favour of this attitude, and 51.53% of the doctors share this opinion. Certain groups supported the question of active euthanasia more clearly than others. These were young participants in the investigation, first-time employees, nondenominational interviewees, those who are dissatisfied with their job situation, who are from the newly-formed German states, and who are divorced. The attitudes expressed by all these people originate in many different motives: thoughts about the patients, aspects concerning jobs, and personal aspects had an influence on results of the investigation. This single investigation which was restricted to patients suffering from coma vigil and to employees of the public health service, does not prove that the total population has generally changed its attitude toward active euthanasia. It is impossible to justify the necessity of new laws about euthanasia based on the above results.

Enhancement of a 252Cf-based neutron beam via subcritical multiplication for neutron capture therapy.

Previous studies indicated that an epithermal-neutron beam based on bare 252Cf is not feasible for neutron capture therapy (NCT). It was reported that a clinically useful epithermal-neutron beam requires a minimum of 1.0 g of 252Cf, which is more than twice the US current annual supply. However, it was reasoned that the required quantity of 252Cf could be dramatically reduced when used with a subcritical multiplying assembly (SMA). This reasoning is based on the assumption that the epithermal-neutron beam intensity for NCT is directly proportional to the fission neutron population, and that the neutron multiplying factor of the SMA can be estimated by 1/(1 - k(eff)). We have performed detailed Monte Carlo calculations to investigate the validity of the above reasoning. Our results show that 1/(1 - k(eff)) grossly overestimates the beam enhancement factor for NCT. For example, Monte Carlo calculations predict a beam enhancement factor of 6.0 for an optimized SMA geometry with k(eff) = 0.968. This factor is much less than 31 predicted by 1/(1 - k(eff)). The overestimation is due to the fact that most of the neutrons produced in the SMA are self-shielded, whereas self-shielding is negligible in a bare 252Cf source. Since the beam intensity of a 0.1 g 252Cf with the optimized SMA enhancement is still more than an order of magnitude too low compared to the existing reactor beams, we conclude that the enhancement via an SMA for a 252Cf-based epithermal-neutron beam is inadequate for NCT.

Lipid peroxidation and the expressional regulation of the heat-shock response during ischemia-reperfusion of rat kidney.

Because of the continuing shortage of donor organs, 'marginal kidneys' are increasingly being used. The purpose of our experiments was to characterize the extent of lipid peroxidation after ischemia-reperfusion (IR) injury in rat kidney, to analyze the expressional regulation of the heat-shock response and now to discuss the clinical application of these results. After ischemia, xanthine oxidase (XO) is thought to be the main oxygen radical-generating system and malondialdehyde (MDA) is considered to be a marker of LPO. In young rats (10 weeks) a unilateral warm ischemia of 40 and 60 min duration with subsequent reperfusion up to 1 h was conducted. Beside the 'footprints' of oxidative stress, the cytosolic antioxidative capacity, expressed as superoxide anion (SOA) scavenging capacity, was investigated. There was only a moderate and transient increase of renal MDA 5 and 10 min after the onset of reoxygenation (133.57/70.67 and 97.84/91.57 vs. 49.47 nmol/g wet weight (ww) in preischemic controls). ATP breakdown (to 83/65 from 2,947 nmol/g ww) with consecutive accumulation of hypoxanthine (up to 1,105 nmol/g ww) at the end of the ischemic period and the subsequent rapid decline of hypoxanthine by XO during reperfusion were used for an assessment of the SOA-generating capacity of these kidneys. Only 1/25-1/50 of the kidney cytosol was able to scavenge the whole amount of SOA generated by the total XO activity of rat kidney. Thus, it could be analytically and stoichiometrically shown that after IR there is only a moderate oxidative stress in kidneys of young rats; this is due to their high SOA-scavenging capacity compared to their SOA-generating ability. We investigated the time course of HSP70-1 and -2 mRNA expression and its relation to cellular ATP levels in renal cortex after different periods of unilateral warm renal ischemia (10-60 min) and reperfusion (up to 60 min) in 10-week-old male Wistar rats, since IR is known to cause induction of both genes. Immediately after ischemia there was a significant induction of both HSP70i genes. While HSP70-1 expression constantly increased (up to 4-fold) during reperfusion, even to a higher extent with prolongation of ischemia, HSP70-2 mRNA - generally being expressed on a far lower level than HSP70-1 mRNA - was strongly induced (3-fold) during reperfusion only after brief periods (10 min) of ischemia. Cellular ATP levels rapidly dropped down to 5% with ischemia and the pattern of recovery during reperfusion significantly depended on the duration of the ischemic period thus showing a good relation to the heat-shock (protein) gene expression. We conclude that the HSP70-2 is the more sensitive gene with a lower threshold activation by mild injury, while the HSP70-1 gene mediates the big response of HSP induction after severe injury. Thus, the measurement of the cytosolic antioxidative capacity and the differential expression of HSP70-1 and -2 mRNA could be promising clinical tools to assess the donor viability.

Comparison and application of a novel genotyping method, semiautomated primer-specific and mispair extension analysis, and four other genotyping assays for detection of hepatitis C virus mixed-genotype infections.

To date the true prevalence of hepatitis C virus (HCV) mixed-genotype infections has not been established mainly because currently available methods are not suitable for the detection of mixed genotypes in a viral population. A novel semiautomated genotyping method, primer-specific and mispair extension analysis (S-PSMEA), which is more reliable than other genotyping assays was developed for detection of HCV mixed-genotype infections. A genotype present at levels as low as 0.8% in a defined mix of HCV genotypes was detected, showing a 20-fold increase in sensitivity over that of direct DNA sequencing. A total of 434 HCV isolates were genotyped and analyzed for a comparative study of the accuracy between S-PSMEA and four current genotyping methods. The results showed that viruses in approximately 40% of the samples from this group determined to be infected with mixed genotypes by S-PSMEA were undetected by direct DNA sequencing due to its low sensitivity. Type-specific PCR, line probe assay, and restriction fragment length polymorphism analysis performed poorly, being able to identify only 38.5, 16.1, and 15.4% of mixed-genotype infections, respectively, that were detected by direct DNA sequencing. The prevalence of mixed-genotype infections detected by S-PSMEA was 7.9% (12 of 152 donors) among HCV-infected blood donors, 14.3% (15 of 105) among patients with chronic hepatitis C, and 17.1% (6 of 36) among thalassemia patients who had received multiple transfusions. The data lead us to conclude that HCV mixed-genotype infections are more common than previously estimated and that S-PSMEA may be the method of choice when detection of genotypes present at low levels in mixed-genotype infections is required due to its higher level of sensitivity.

Prevalence of GBV-C/hepatitis G virus viremia and anti-E2 in Canadian blood donors.

BACKGROUND AND OBJECTIVES: GB virus C (GBV-C)/hepatitis G virus (HGV) is a recently recognized parenterally and sexually transmitted agent. The prevalence of GBV-C/HGV markers in Canadian blood donors has not been previously studied and was therefore determined. MATERIALS AND METHODS: Blood donors [identity unlinked (IU), short-term temporarily deferred (STTD) and autologous groups] and donor samples with antibodies to hepatitis C (anti-HCV) or hepatitis B core were tested for GBV-C/HGV RNA and for antibodies to E2 antigen (anti-E2). RESULTS: GBV-C/HGV RNA was found in 1.1% and anti-E2 in 7.3% of the combined IU/STTD donor group. Viremia was much more common in anti-HCV-positive samples (12.5%); anti-E2 was present in >50% of this group. In the STTD group, female gender was significantly associated with viremia. CONCLUSION: GBV-C/HGV infection is relatively common in Canadian donors, and a small proportion are viremic. The association of female gender and viremia was unexpected. Further study is needed to clarify the epidemiology and natural history of GBV-C/HGV infection.

Evidence for only a moderate lipid peroxidation during ischemia-reperfusion of rat kidney due to its high antioxidative capacity.

The extent of lipid peroxidation after ischemia-reperfusion (I-R) injury in rat kidney has been controversial. After I, xanthine oxidase (XO) is thought to be the main oxygen radical-generating system and malondialdehyde (MDA) is considered to be a marker of lipid peroxidation (LPO). In young rats (10 weeks old) a unilateral warm I of 40 and 60 min duration with subsequent R up to 1 h was conducted. Beside the "footprints" of oxidative stress, the cytosolic antioxidative capacity, expressed as superoxide anion (SOA) scavenging capacity, and the renal catalase were also investigated. There was only a moderate and transient increase of renal MDA 5 and 10 min after the onset of reoxygenation (133.57/70. 67 and 97.84/91.57 vs. 49.47 nmol/g ww in preischemic controls). ATP breakdown (to 83/65 from 2947 nmol/g ww) with consecutive accumulation of hypoxanthine (up to 1105 nmol/g ww) at the end of ischemic period and the subsequent rapid decline of hypoxanthine by XO during reperfusion were used for an assessment of the SOA-generating capacity of these kidneys. Superoxide dismutase (SOD) activity, glutathione (GSH) and the high activity of catalase (18000 U/g ww) remained nearly unchanged during R. Only 1/25-1/50 of the kidney cytosol was able to scavenge the whole amount of SOA generated by the total XO activity of rat kidney. Thus, it could be analytically and stoichiometrically shown that after IR there is only a moderate oxidative stress in kidneys of young rats; this is due to their high SOA-scavenging capacity compared with their SOA-generating ability.

Plasmin converts factor X from coagulation zymogen to fibrinolysis cofactor.

Known anticoagulant pathways have been shown to exclusively inhibit blood coagulation cofactors and enzymes. In the current work, we first investigated the possibility of a novel anticoagulant mechanism that functions at the level of zymogen inactivation. Utilizing both clotting and chromogenic assays, the fibrinolysis protease plasmin was found to irreversibly inhibit the pivotal function of factor X (FX) in coagulation. This was due to cleavage at several sites, the location of which were altered by association of FX with procoagulant phospholipid (proPL). The final products were approximately 28 and approximately 47 kDa for proPL-bound and unbound FX, respectively, which did not have analogues when activated FX (FXa) was cleaved instead. We next investigated whether the FX derivatives could interact with the plasmin precursor plasminogen, and we found that plasmin exposed a binding site only on proPL-bound FX. The highest apparent affinity was for the 28-kDa fragment, which was identified as the light subunit disulfide linked to a small fragment of the heavy subunit (Met-296 to approximately Lys-330). After cleavage by plasmin, proPL-bound FX furthermore was observed to accelerate plasmin generation by tissue plasminogen activator. Thus, a feedback mechanism localized by proPL is suggested in which plasmin simultaneously inhibits FX clotting function and converts proPL-bound FX into a fibrinolysis cofactor. These data also provide the first evidence for an anticoagulant mechanism aimed directly at the zymogen FX.

No association between hepatitis C and B-cell lymphoma.

Chronic viral infection has been implicated in the pathogenesis of B-cell lymphoma, and hepatitis C virus (HCV) infects lymphocytes. Chronic infection with HCV may result in B-cell proliferation. Individuals infected with hepatitis C are often co-infected with the RNA virus GB virus type C. Studies from Europe where hepatitis C infection is more common than in North America have shown a high prevalence of hepatitis C infection in patients with B-cell lymphoma. The aim of this study was to establish the prevalence of HCV and GBV-C infection in patients with B-cell lymphoma in an area of low HCV prevalence. One hundred patients with B-cell lymphoma (10 high grade, 46 intermediate grade, and 44 low grade) and 100 controls with nonhematological malignancies were studied. Serum was analyzed for HCV antibodies by third generation enzyme-linked immunosorbant assay, and HCV RNA and GBV-C RNA was analyzed by reverse transcriptase PCR. None of the controls or lymphoma patients had antibodies to HCV. HCV RNA was undetected in 60 out of 100 lymphoma patients tested. GBV-C RNA was detected in the serum of 5 out of 100 (5%) of lymphoma patients and in 3 out of 100 (3%) controls. Hepatitis C and GBV-C are, therefore, unlikely to play a major role in the pathogenesis of B-cell lymphoma in North America.

Primer specific and mispair extension analysis (PSMEA) as a simple approach to fast genotyping.

A simple method, primer specific and mispair extension analysis (PSMEA) with pfu DNA polymerase was developed for genotyping. PSMEA is based on the unique properties of 3'-->5' exonuclease proofreading activity. In the presence of an incomplete set of dNTPs, pfu was found to be extremely discriminative in nucleotide incorporation and proofreading at the initiation step of DNA synthesis, completely preventing primer extension when mispair(s) are found adjacent to the 3'-end of the primer. This has allowed us to accurately detect nucleotide variations, deletions and insertions for fast genotyping.

Comparison of continuous spinal with combined spinal-epidural anesthesia using plain bupivacaine 0.5% in trauma patients.

We investigated the efficacy and complications of microcatheter spinal anesthesia (CSA) in comparison to a combined spinal-epidural technique (CSE) using plain bupivacaine 0.5%. Sixty trauma patients randomly received either CSA using a 22-gauge Sprotte needle and a 28-gauge microcatheter or CSE after insertion of a 22-gauge epidural catheter through an 18-gauge Tuohy needle followed by dural puncture with a 25-gauge pencil-point needle inserted through the backeye of the Tuohy needle. An initial subarachnoid bolus of 2 mL of plain bupivacaine 0.5% was injected. If analgesia did not reach T12 within 20 min, supplemental bupivacaine was injected either intrathecally or epidurally up to a maximum of 5 mL in the CSA group or 16 mL in the CSE group. Mean arterial blood pressure, heart rate, and analgesic levels were recorded. On postoperative Day 4, patients were interviewed for postanesthetic complaints. Technical problems were more frequent in the CSE group than in the CSA group (47% vs 13%). Performance of anesthesia was faster (8 +/- 3 vs 15 +/- 8 min) and the total dose of bupivacaine lower (3.2 +/- 1.0 vs 9.7 +/- 5 mL) in patients who received CSA. The incidence of hypotension did not differ significantly. However, more patients in the CSE group were treated for bradycardia (4 vs 0). The number of patients suffering from postdural puncture headache was comparable in both groups, but there were more patients with lower back pain in the CSE group (8 vs 2). In conclusion, our data suggest that microcatheter CSA is not associated with an increased rate of complication in patients with lower limb fractures.

[Life-threatening swelling of the tongue in antihypertensive therapy with ACE inhibitors]. / Lebensbedrohliche Zungenschwellung bei antihypertensiver Therapie mit ACE-Hemmern.

Although generally showing a low incidence of side effects, inhibitors of angiotensin-converting enzyme (ACE) may in rare cases induce angioedemas, mainly located in the oro-facial area and larynx. The interval between the beginning of the ACE inhibitor therapy and the occurrence of such angioedemas may range from a few hours to a few years. Here, the case of a 53-year-old man with massive swelling of the tongue after dental surgery is presented who had started with ACE inhibitor therapy only 24 h before. At admission to the clinic, obstruction of the upper airway due to the tongue swelling had already progressed so far that fiberoptic intubation was necessary. Additionally, the patient was treated with corticosteroids, antihistaminics and epinephrine, avoiding any further administration of the ACE inhibitor. The swelling resolved within 48-72 h. Dentists and physicians should take into consideration this potential side effect in patients treated with ACE inhibitors.

Autoproteolysis or plasmin-mediated cleavage of factor Xaalpha exposes a plasminogen binding site and inhibits coagulation.

Blood coagulation factor Xa (FXa) has recently been shown to function as a plasminogen receptor in the presence of procoagulant phospholipid (phosphatidylserine; PS) and Ca2+. In the current work, the possible effect of autoproteolytic and plasmin-mediated cleavage of FXa on complex formation with plasminogen was investigated. 125I-plasminogen binding to derivatives of FXa electrotransferred to polyvinylidene difluoride revealed that the autoproteolytic conversion of FXaalpha to FXabeta was required for the expression of a plasminogen binding site. In the presence of PS and Ca2+, plasmin was shown to convert FXaalpha to a FXabeta-like species at least 3 orders of magnitude faster than the autoproteolytic mechanism. This also resulted in the exposure of a plasminogen binding site. Further processing by plasmin generated a fragment (33 kDa) due to cleavage at Gly331 in the FXa heavy chain. Production of this species enhanced apparent plasminogen binding compared with FXabeta and resulted in the loss of FXa amidolytic and clotting activity. In the absence of either PS or Ca2+, the plasmin-mediated fragmentation of FXaalpha was altered to include a FXabeta-like molecule and a species (40 kDa) with intact beta-heavy chain disulfide linked to a COOH-terminal fragment of the light chain starting at Tyr44. Neither of these products was observed to interact with plasminogen. The 40-kDa species had amidolytic activity comparable with FXaalpha but inhibited clotting activity. Cumulatively the data provide the first evidence for a functional difference between the FXa subforms and suggest a mechanism where autoproteolysis and plasmin-mediated cleavage modulate the function of FXaalpha from a procoagulant enzyme to a profibrinolytic plasminogen receptor.

Kinetics of blood coagulation factor Xaalpha autoproteolytic conversion to factor Xabeta. Effect on inhibition by antithrombin, prothrombinase assembly, and enzyme activity.

Autoproteolysis of blood coagulation factor Xa (FXa) results in the excision of a 4-kDa fragment (beta-peptide) from the intact subform, factor Xaalpha (FXaalpha), to yield factor Xabeta (FXabeta). In the preceding paper, we showed that generation of FXabeta leads to expression of a plasminogen binding site. FXabeta may consequently participate in fibrinolysis; therefore, the timing of subform conversion compared with thrombin production is important. In the current study we evaluated the kinetics of FXabeta generation, which showed that autoproteolysis of FXaalpha followed a second order mechanism where FXaalpha and FXabeta behaved as identical enzymes. Rate constants of 9 and 172 M-1 s-1 were derived, respectively, in the absence and presence of FXaalpha binding to procoagulant phospholipid. Under identical conditions the latter is estimated to be 6 orders of magnitude slower than thrombin generation by prothrombinase. Since heparin binding and prothrombin recognition have been previously attributed to a region of FXaalpha proximal to the beta-peptide, functional comparisons were conducted using homogeneous and stabilized preparations of FXaalpha and FXabeta. Comparisons included 1) the recognition of small substrates; 2) the rate of interaction with antithrombin/heparin; 3) the assembly of prothrombinase; and 4) the activation of prothrombin by prothrombinase. Although the beta-peptide neighbors a probable functional region in FXaalpha, conversion to FXabeta was not observed to influence these functions. The data support a model where FXaalpha is predominantly responsible for thrombin generation and where slow conversion to FXabeta coordinates coagulation and the initiation of fibrinolysis at sites of prothrombinase assembly.