Registered Replication Report: Rand, Greene, and Nowak (2012).

In an anonymous 4-person economic game, participants contributed more money to a common project (i.e., cooperated) when required to decide quickly than when forced to delay their decision (Rand, Greene & Nowak, 2012), a pattern consistent with the social heuristics hypothesis proposed by Rand and colleagues. The results of studies using time pressure have been mixed, with some replication attempts observing similar patterns (e.g., Rand et al., 2014) and others observing null effects (e.g., Tinghög et al., 2013; Verkoeijen & Bouwmeester, 2014). This Registered Replication Report (RRR) assessed the size and variability of the effect of time pressure on cooperative decisions by combining 21 separate, preregistered replications of the critical conditions from Study 7 of the original article (Rand et al., 2012). The primary planned analysis used data from all participants who were randomly assigned to conditions and who met the protocol inclusion criteria (an intent-to-treat approach that included the 65.9% of participants in the time-pressure condition and 7.5% in the forced-delay condition who did not adhere to the time constraints), and we observed a difference in contributions of -0.37 percentage points compared with an 8.6 percentage point difference calculated from the original data. Analyzing the data as the original article did, including data only for participants who complied with the time constraints, the RRR observed a 10.37 percentage point difference in contributions compared with a 15.31 percentage point difference in the original study. In combination, the results of the intent-to-treat analysis and the compliant-only analysis are consistent with the presence of selection biases and the absence of a causal effect of time pressure on cooperation.

A comparison of the Enk Fiberoptic Atomizer Set(™) with boluses of topical anaesthesia for awake fibreoptic intubation.

We compared the Enk Fiberoptic Atomizer Set(™) with boluses of topical anaesthesia administered via the working channel during awake fibreoptic tracheal intubation in 96 patients undergoing elective surgery. Patients who received topical anaesthesia via the atomiser, compared with boluses via the fibreoptic scope, reported a better median (IQR [range]) level of comfort: 1 (1-3 [1-10]) vs. 4 (2-6 [1-10]), p < 0.0001; experienced a reduced total number of coughs: 6 (3-10 [0-34]) vs. 11 (6-13 [0-25]), p = 0.0055; and fewer distinct coughing episodes: 7% vs. 27% respectively, p = 0.0133. The atomiser technique was quicker: 5 (3-6 [2-12]) min vs. 6 (5-7 [2-15]) min, p = 0.0009; and required less topical lidocaine: 100 mg (100-100 [80-160]) vs. 200 mg (200-200 [200-200]), p < 0.0001. Four weeks after nasal intubation, the incidence of nasal pain was less in the atomiser group compared with the control group (8% vs. 50%, p = 0.0015). We conclude that the atomiser was superior to bolus application for awake fibreoptic tracheal intubation.

Significance of platelet function diagnostics for clarification of suspected battered child syndrome.

The manifestation of an unclear bleeding tendency in childhood calls for an extended coagulation work-up, particularly when a battered child syndrome is suspected and typical concomitant injuries are absent. The chosen diagnostic tests should be able to detect the presence of relatively common coagulation defects such as von Willebrand syndrome or hemophilia, but also rare diseases such as inherited thrombocytopathies. The PFA-100® test does not help to provide a definite diagnosis especially in cases of mild inherited thrombocytopathies, since in most cases the PFA-100® test results are normal. For this purpose, specific platelet function testing is needed. However, the methods are only available in some coagulation laboratories. Also, other limitations need to be taken into consideration such as pre-analytical problems and difficulties in the interpretation of test results especially in infants. We present two cases that were diagnosed with an aspirin-like defect as an inherited thrombocytopathy, even though their PFA-100 closure times were within the normal range. Based on pathological findings in the platelet aggregometry test, this diagnosis could be made.

Comprehension of climate change and environmental attitudes across the lifespan.

Given the coincidence of the demographic change and climate change in the upcoming decades the aging voter gains increasing importance in climate change mitigation and adaptation processes. It is generally assumed that information status and comprehension of complex processes underlying climate change are prerequisites for adopting pro-environmental attitudes and taking pro-environmental actions. In a cross-sectional study, we investigated in how far (1) environmental knowledge and comprehension of feedback processes underlying climate change and (2) pro-environmental attitudes change as a function of age. Our sample consisted of 92 participants aged 25-75 years (mean age 49.4 years, SD 17.0). Age was negatively related to comprehension of system structures inherent to climate change, but positively associated with level of fear of consequences and anxiousness towards climate change. No significant relations were found between environmental knowledge and pro-environmental attitude. These results indicate that, albeit understanding of relevant structures of the climate system is less present in older age, age is not a limiting factor for being engaged in the complex dilemma of climate change. Results bear implications for the communication of climate change and pro-environmental actions in aging societies.

Therapy refractory menorrhagia as first manifestation of Hermansky-Pudlak syndrome.

INTRODUCTION: Oculocutaneous albinism (OCA) in combination with a platelet function defect caused by a disturbed release reaction from platelet δ-granules (storage pool defect - SPD) is typical for the autosomal recessive inherited Hermansky-Pudlak syndrome (HPS). CASE REPORT: A girl (age: 13 years) with OCA was hospitalized with transfusion-requiring menorrhagia. The suspicion of HPS was confirmed by results of lumi-aggregometry. Suspecting a disorder in primary haemostasis treatment with tranexamic acid (10 mg/kg body weight every 8 h i. v.), desmopressin (0.3 µg/kg body weight every 8 to 12 h) and hormonal therapy (norethisterone) was started but the menorrhagia persisted. Clinical response was finally achieved by a single injection of 100 µg/kg body weight recombinant factor VIIa (rFVIIa). CONCLUSION: The diagnosis of HPS should be suspected in patients with OCA and bleeding symptoms and is confirmed by the proof of SPD. In case of absent clinical response to desmopressin the application of rFVIIa should be considered. Hormones and antifibrinolytics are useful options in the treatment of extensive menorrhagia.

Evaluation of desmopressin effect on primary haemostasis in pediatric patients with aspirin-like defect as hereditary thrombocytopathy.

OBJECTIVES: Despite about 3 decades of clinical experience with the therapy of inherited thrombocytopathies (HTP) with desmopressin (DDAVP) the mechanisms of haemostatic effects of DDAVP in these diseases remain unclear. Therefore platelet function diagnostics was carried out in whole blood (WB) from children with aspirin-like defect as one of the clinically mild forms of HTP after DDAVP administration. DESIGN AND METHODS: 11 children (age range: 3-16 years) were treated with DDAVP i.v. (0.3 µg/kg as short infusion). Before, after 120, and 240 min of DDAVP administration the following parameters were measured: platelet aggregation (PA) and ATP release induced by ADP, collagen, ristocetin and thrombin; PFA-100 closure times (CT), factor VIII activity (FVIII:C), Von Willebrand factor antigen (VWF:Ag), collagen binding activity (VWF:CB) and blood count. RESULTS: PA, ATP release and blood count were not influenced by DDAVP administration. PFA-100 CTs were markedly reduced at 120 and 240 min after DDAVP, respectively. FVIII:C, VWF:Ag and VWF:CB were increased after 120 min. CONCLUSION: The DDAVP-induced improvement of primary haemostasis in patients with aspirin-like defect is mainly due to the marked increase of the VWF. For the evaluation of the clinical effect of DDAVP administration in patients with aspirin-like defect the investigation of a larger group of patients is needed.

Platelet function in obese children and adolescents.

UNLABELLED: Platelet hyperaggregability contributes to thromboembolic events of obesity in adulthood. In obese children hyperaggregability was described in platelet rich plasma. We investigated platelet aggregation in children with obesity and lipometabolic disorders in whole blood. PATIENTS, MATERIAL, METHODS: Specimens from patients with overweight (n = 35), hypercholesterolaemia and normal weight (n = 5), overweight plus combined lipometabolic disorder (n = 5) and healthy controls (n = 20) were investigated. Aggregation and ATP release were induced by ADP (20 µmol/l), collagen (1 µg/ml) and thrombin (0.5 U/ml) using a lumiaggregometer. RESULTS: Overweight children and normal weight patients with hypercholesterolaemia exhibited no significant differences in platelet aggregation compared to controls. Contrastingly, in patients with obesity plus lipometabolic disorder the aggregation rate was significantly higher (p < 0.05) suggesting a hyperaggregable state. CONCLUSION: Obviously in obese children a hypercoagulable state exists and the slight hyperaggregability observed in whole blood in this cohort might contribute to that. Any effort should be undertaken to avoid obesity in children especially in those countries where the prevalence of obesity in childhood is continuously increasing.

Validity of species status of the parasitic mite Otodectes cynotis.

A combined molecular and phenotypic approach was used to determine whether ear mites of the genus Otodectes (Acari: Psoroptidae) belong to a single species. The second internal transcribed spacer (ITS 2) of the rDNA of 16 isolates from 11 cats, two dogs, one arctic fox and two ferrets originating from four different continents was characterized. In addition, mites from dog, cat and arctic fox were investigated morphologically. Sequence comparisons revealed five different, but closely related genotypes which did not segregate according to host species or geographical origin. Morphologically, mites of the three host species did not differ significantly in their body or leg sizes. These investigations support the view that ear mites of the genus Otodectes from different hosts and geographical origins belong to a single species, Otodectes cynotis (Hering).

Evaluation of a topical solution containing 65% permethrin against the sandfly (Phlebotomus perniciosus) in dogs.

A topical solution containing 65% permethrin (EXspot, Schering-Plough Animal Health Corp., Union, NJ) was evaluated for repellency and insecticidal activity against the phlebotomine sandfly, Phlebotomus perniciosus (Diptera: Psychodidae). Four dogs were sedated and individually exposed to 100 female P. perniciosus inside a cage. Two of the dogs were treated on Day 0 with the permethrin solution by application of a 2-ml dose directly on the skin along the dorsal midline from the dorsal scapular area to the base of the tail. Two dogs remained untreated as controls. On Days -8, 0, 7, 14, 21, 28, 35, 49, 70, and 91, sandfly landings were recorded during the first 5 minutes of an insect exposure period and insect feeding and mortality rates were recorded after 1 hour of exposure. The number of sandfly landings was moderately decreased, and insect feeding rates decreased to zero by Day 7. From Day 14 through Day 28, the number of insect feedings was reduced from control values by greater than 90%. These findings indicate the permethrin spot-on provided a considerable repellent effect. Insect mortality rates remained very high (about 61%) during the 5 weeks after application, although insecticidal effects persisted for more than 7 weeks in total. These data suggest that monthly application of this topical solution of permethrin from the end of spring until the beginning of fall may protect dogs from the majority of sandfly bites. Further investigation with greater numbers of animals is warranted; however, results of this preliminary investigation indicate that a leishmaniasis control program that incorporates the strategic use of this product on dogs can be expected to decrease the number of infected animals and the incidence of the disease among the canine population.

Double duplex invasion by peptide nucleic acid: a general principle for sequence-specific targeting of double-stranded DNA.

Pseudocomplementary PNAs containing diaminopurine.thiouracil base pairs have been prepared and are shown to bind with high specificity and efficiency to complementary targets in double-stranded DNA by a mechanism termed "double duplex invasion" in which the duplex is unwound and both DNA strands are targeted simultaneously, each by one of the two pseudocomplementary peptide nucleic acids (PNAs). On the basis of our results we predict that (for decameric targets) more than 80% of all sequences can be targeted by straightforward Watson-Crick base pairing by using this approach in its present form. Targeting of pseudocomplementary PNAs to the promoter of the T7 phage RNA polymerase effectively inhibits transcription initiation. These results have important implications in the development of gene therapeutic agents as well as for genetic diagnostic and molecular biology applications.

Ubiquitous expression of marker transgenes in mice and rats.

The ability to unambiguously mark a cell's genotype is essential for studies in which genetically distinct cell populations must be distinguished from one another in vivo. One approach to this challenge has been the creation of transgenic mice expressing a transgene marker that is easily detectable, with no background staining. Multiple transgenic mouse strains bearing constructs with different combinations of promoter elements and coding sequences have been described, each with its own advantages and limitations. In this report we describe the use of an 800-bp promoter fragment isolated from the beta(geo) integration site in ROSA26 mice to target expression of two marker genes. We demonstrate that the ROSA26 promoter directs ubiquitous expression of human placental alkaline phosphatase and enhanced green fluorescent protein during embryonic and postnatal development in mouse and rat. We further demonstrate the general utility of these transgenes for marking donor cells in transplantation studies.

Infection of sand flies by humans coinfected with Leishmania infantum and human immunodeficiency virus.

To determine the role that Leishmania infantum/human immunodeficiency virus (HIV) coinfected patients could play in the epidemiology of visceral leishmaniasis (VL), we applied direct xenodiagnosis of VL in this study to test the infectivity of six coinfected patients to colonized Phlebotomus perniciosus. All patients proved to be infective for the sand flies. The infectivity of patients who had still not received specific treatment for VL was inversely proportional to their absolute CD4+ T lymphocyte cell count. It has been proven that P. perniciosus can acquire and allow the development of L. infantum by feeding on L. infantum/HIV coinfected patients. Since this sand fly is an important vector of VL in southern Europe, a new natural anthroponotic cycle could be considered in the epidemiology of L. infantum/HIV coinfection. The design of leishmaniasis control programs and the management of coinfected individuals should take these findings into account.

Fluorescein-conjugated lysine monomers for solid phase synthesis of fluorescent peptides and PNA oligomers.

Fluorescein ethyl ester, I, was used to prepare the fluorescent mixed ester/ether 6-O-(carboxymethyl)-fluorescein ethyl ester, III. Conjugation of III to the epsilon-amino group of alpha-N-Boc-L-lysine, via the N-hydroxysuccinimde ester, IV, gave the Boc-protected fluorescein-conjugated lysine monomer V. Removal of the Boc group, followed by reaction with Fmoc chloride, gave the Fmoc-protected monomer, VI (Figure 1). These Boc- and Fmoc-protected fluorescein-conjugated lysines were readily incorporated into peptides and PNA oligomers during solid phase synthesis to give fluorescent products. Mass spectroscopy and UV studies showed that the fluorophore remains unchanged during solid phase synthesis. In contrast to fluorescein, the photophysical properties of these derivatives are pH independent from pH 3 to 8, with a molar absorption coefficient, epsilon max 456, of 2.9 x 10(4) M-1 cm-1 and fluorescence quantum yield, phi f, of 0.18.

Transgenic hepatocarcinogenesis in the rat.

Although transgenic hepatocarcinogenesis has been accomplished in the mouse with a number of genetic constructs targeting the oncogene to expression primarily in the liver, no example of this process has yet been developed in the rat. Because our understanding of the multistage nature of hepatocarcinogenesis is most advanced in the rat, we have developed a strain of transgenic rats carrying the promoter-enhancer sequences of the mouse albumin gene linked 5' to the simian virus-40 T antigen gene. A line of transgenic rats bearing this transgene has been developed from a single founder female. Five to six copies of the transgene, possibly in tandem, occur within the genome of the transgenic animals, which are maintained by heterozygous matings. Livers of transgenic animals are histologically normal after weaning; at 2 months of age, small foci of vacuolated cells appear in this organ. By 4 months of age, all animals exhibit focal lesions and nodules consisting primarily of small basophilic cells, many of which exhibit considerable cytoplasmic vacuolization. Mating of animals each bearing the transgene results in rats with a demyelinating condition that develops acutely in pregnant females and more chronically in males. Ultrastructural studies of these cells indicate that the vacuoles contain substantial amounts of glycogen, with the cells resembling hepatoblasts. Malignant neoplasms with both a glandular and a hepatoblastoma/hepatocellular carcinoma pattern arise from the nodules. Enzyme and immunohistochemical studies of all lesions reveal many similarities in gene expression to comparable lesions in rats subjected to chemically induced hepatocarcinogenesis, with certain exceptions. The placental form of glutathione-S-transferase is absent from all lesions in the transgenic animal, as is the expression of connexin 32. A significant number of lesions express serum albumin, and many, but not all, exhibit the T antigen. Lesions expressing the T antigen also contain stainable amounts of the p53 gene product; by contrast, normal hepatocytes express only very low levels of the T antigen within their nuclei and no demonstrable p53. All of the animals develop hepatic lesions, and approximately one-third also develop adenomas and carcinomas derived from the islet cells of the pancreas. Although there are differences in the morphology, biology, and genetic expression in early and late hepatic lesions in this strain of transgenic rat, many similarities also occur, making this a potential model system with which to study the interactions of environmental factors with a genetic program for hepatocarcinogenesis.

Tumorigenicity by human papillomavirus type 16 E6 and E7 in transgenic mice correlates with alterations in epithelial cell growth and differentiation.

The human papillomavirus type 16 (HPV-16) E6 and E7 oncogenes are thought to play a role in the development of most human cervical cancers. These E6 and E7 oncoproteins affect cell growth control at least in part through their association with and inactivation of the cellular tumor suppressor gene products, p53 and Rb. To study the biological activities of the HPV-16 E6 and E7 genes in epithelial cells in vivo, transgenic mice were generated in which expression of E6 and E7 was targeted to the ocular lens. Expression of the transgenes correlated with bilateral microphthalmia and cataracts (100% penetrance) resulting from an efficient impairment of lens fiber cell differentiation and coincident induction of cell proliferation. Lens tumors formed in 40% of adult mice from the mouse lineage with the highest level of E6 and E7 expression. Additionally, when lens cells from neonatal transgenic animals were placed in tissue culture, immortalized cell populations grew out and acquired a tumorigenic phenotype with continuous passage. These observations indicate that genetic changes in addition to the transgenes are likely necessary for tumor formation. These transgenic mice and cell lines provide the basis for further studies into the mechanism of action of E6 and E7 in eliciting the observed pathology and into the genetic alterations required for HPV-16-associated tumor progression.

Citrate and calcium secretion in the pure human pancreatic juice of alcoholic and nonalcoholic men and of chronic pancreatitis patients.

Citrate, calcium and protein have been estimated in pure pancreatic juice after a secretin and a CCK injection in 4 patients presenting with alcoholic calcified pancreatitis (ACP), 10 controls without evidence of pancreatic disease, drinking more than 130 g alcohol/day, and 10 controls without evidence of pancreatic disease, drinking less than 20 g alcohol/day. Citrate is normally secreted in the pancreatic juice and this secretion increases in parallel with protein after CCK injection. Citrate secretion is significantly decreased in the two alcoholic groups. Calcium secretion is increased in the ACP, and reasons are presented to suggest that this may be due to lesions of the ducts. These modifications could play a role in the formation of pancreatic stones which are mostly built up of calcium carbonate.

Duodenal total and ionised calcium secretion in normal subjects, chronic alcoholics, and patients with various stages of chronic alcoholic pancreatitis.

Previous studies have shown increased secretion of total calcium in the duodenal juice of patients with chronic alcoholic pancreatitis compared with healthy subjects. In order to get more detailed information on calcium secretion and pancreatic stone formation in chronic alcoholic pancreatitis, ionised and total calcium concentrations were determined in the duodenal juice of normal subjects, chronic alcoholics, and patients with various stages of chronic alcoholic pancreatitis. Total calcium secretion was in agreement with previously published data. Chronic alcoholics presented a significant increase of ionised calcium. In the course of pancreatitis all calcium fractions increased progressively revealing highest concentrations in patients with severe exocrine insufficiency. In non-calcified and calcified pancreatitis all calcium fractions were identical. It is suggested that the increase of ionised calcium originates from serum ionised calcium passing by diffusion into the damaged pancreatic duct system.

Calcium binding to the "stone protein" isolated from pancreatic stones of patients with chronic calcified pancreatitis.

Calcium binding to a new secretory protein present in all layers of human pancreatic stones has been studied. This protein with a molecular mass of 13,500 Da has about 4 equivalent binding sites for calcium. Maximum calcium binding was independent of pH and temperature, but the affinity of calcium to this protein increased with decreasing pH and increasing temperature, revealing maximum affinity at pH 7.3 and 37 degrees C. It is concluded that the calcium affinity could explain the presence of the protein in all layers of human pancreatic stones.