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Microcirculatory dysfunction, hypoxia, and inflammation are considered to be central in the pathogenesis of sepsis-induced acute kidney injury (AKI). In this experimental study, we hypothesized that extracorporeal removal of inflammatory cytokines by hemoadsorption (HA) therapy may mitigate renal injury associated with sepsis-induced AKI. To this end, we investigated renal microcirculatory oxygenation and perfusion, oxygen consumption, lactate, systemic hemodynamic variables, tubular cell integrity, inflammatory mediators, and kidney function in a rat model of septic AKI elicited by endotoxin infusion. Three groups of rats were investigated on extracorporeal circulation: HA only, LPS, and LPS + HA. Endotoxin infusion reduced cortex microcirculatory oxygenation and raised creatinine and lactate levels. Renal microcirculatory oxygenation, measured by two independent techniques (phosphorescence (µPO 2 ) and spectrophotometry/Doppler (µHbO 2sat and [Formula: see text])), was ameliorated by HA therapy. The renal oxygen consumption, lactate and creatinine levels were restored in the LPS + HA group. A reduced amount of injured tubular cells was found in histological analysis of the kidneys. This experimental study demonstrated an improvement in multiple determinants of kidney oxygenation, damage, and systemic blood perfusion by HA in a clinically relevant rat model of septic AKI. Further studies are needed to optimize and support the clinical use of HA as a renal protective strategy., Competing Interests: Declarations Competing interests Dr. Ince is the CSO of Active Medical BV, Leiden, The Netherlands, a company that provides devices (OxyCam), software (MicroTools), education (Microcirculation Academy), and services related to clinical microcirculation. All other authors declare that they have no conflict of interest., (© 2024. The Author(s).)

Introduction: While secondhand smoke exposure in outdoor spaces has been investigated before, no data on outdoor secondhand smoke exposure have been collected in the Netherlands. Such data could help policymakers gain support for smoke-free outdoor public spaces., Methods: Between May and November 2021, we visited 25 outdoor locations across the Netherlands. At each location, we conducted four measurements with smokers and one measurement without smokers. During each measurement, we counted the number of smokers present and we rated tobacco smell intensity on a five-point scale. Airborne nicotine and 3-ethenylpyridine (3-EP) data were collected through active sampling on thermal desorption tubes. The contents of these tubes were later analyzed using gas chromatography-mass spectrometry. Using linear mixed models, we investigated the association between levels of nicotine and the presence of smokers, the number of smokers, and the intensity of tobacco smell. We also investigated these association with levels of 3-EP., Results: Nicotine levels were higher when smokers were present (B=1.40; 95% CI: 0.69-2.11, p<0.001). For each additional smoker present, we measured higher levels of nicotine (B=0.23; 95% CI: 0.10-0.37, p=0.001). When the smell of tobacco smoke was noted to be stronger by the researchers, higher levels of nicotine were measured through sampling (B=0.85; 95% CI: 0.44-1.26, p<0.001). We found similar results for 3-EP levels., Conclusions: This study showed that both nicotine and 3-EP are useful in quantifying levels of secondhand smoke in various outdoor locations. The level of nicotine exposure outdoors was positively associated with the number of smokers nearby. The intensity of the tobacco smell was also related to nicotine exposure: the stronger the smell of tobacco smoke, the more nicotine was measured in the air., Competing Interests: The authors have completed and submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. The authors declare that they have no competing interests, financial or otherwise, related to the current work. All authors report that since the initial planning of the work J. Bommelé received a joint grant from the Dutch Heart Foundation, Lung Foundation Netherlands, Dutch Cancer Society, Dutch Diabetes Research Foundation and Netherlands Thrombosis Foundation (2.1.19.001), and a grant from the Netherlands Ministry of Health, Welfare and Sport (329845)., (© 2024 Bommelé J. et al.)

To suppress serious influenza infections in persons showing insufficient protection from the vaccines, antiviral drugs are of vital importance. There is a need for novel agents with broad activity against influenza A (IAV) and B (IBV) viruses and with targets that differ from those of the current antivirals. We here report a new small molecule influenza virus inhibitor referred to as CPD A (chemical name: N-(pyridin-3-yl)thiophene-2-carboxamide). In an influenza virus minigenome assay, this non-nucleoside compound inhibited RNA synthesis of IAV and IBV with EC 50 values of 2.3 μM and 2.6 μM, respectively. Robust in vitro activity was noted against a broad panel of IAV (H1N1 and H3N2) and IBV strains, with a median EC 50 value of 0.20 μM, which is 185-fold below the 50% cytotoxic concentration. The action point in the viral replication cycle was located between 1 and 5 h p.i., showing a similar profile as ribavirin. Like this nucleoside analogue, CPD A was shown to cause strong depletion of the cellular GTP pool and, accordingly, its antiviral activity was antagonized when this pool was restored with exogenous guanosine. This aligns with the observed inhibition in a cell-based IMP dehydrogenase (IMPDH) assay, which seems to require metabolic activation of CPD A since no direct inhibition was seen in an enzymatic IMPDH assay. The combination of CPD A with ribavirin, another IMPDH inhibitor, proved strongly synergistic. To conclude, we established CPD A as a new inhibitor of influenza A and B virus replication and RNA synthesis, and support the potential of IMPDH inhibitors for influenza therapy with acceptable safety profile., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Hepatic uptake of triglyceride-rich remnant lipoproteins is mediated by the low-density lipoprotein receptor, a low-density lipoprotein receptor related protein and the heparan sulfate proteoglycan, syndecan-1. Heparan sulfate proteoglycan also mediates low-density lipoprotein receptor degradation by a regulator of cholesterol homeostasis, proprotein convertase subtilisin kexin type 9 (PCSK9), thereby hampering triglyceride-rich remnant lipoproteins uptake. In this study, we investigated the effects of proteinuria on PCSK9, hepatic heparan sulfate proteoglycan and plasma triglyceride-rich remnant lipoproteins. Adriamycin-injected rats developed proteinuria, elevated triglycerides and total cholesterol (all significantly increased). Proteinuria associated with triglycerides and total cholesterol and serum PCSK9 (all significant associations) without loss of the low-density lipoprotein receptor as evidenced by immunofluorescence staining and western blotting. In proteinuric rats, PCSK9 accumulated in sinusoids, whereas in control rats PCSK9 was localized in the cytoplasm of hepatocytes. Molecular profiling revealed that the heparan sulfate side chains of heparan sulfate proteoglycan to be hypersulfated in proteinuric rats. Competition assays revealed sulfation to be a major determinant for PCSK9 binding. PCSK9 partly colocalized with hypersulfated heparan sulfate in proteinuric rats, but not in control rats. Hence, proteinuria induces hypersulfated hepatic heparan sulfate proteoglycans, increasing their affinity to PCSK9. This might impair hepatic triglyceride-rich remnant lipoproteins uptake, causing proteinuria-associated dyslipidemia. Thus, our study reveals PCSK9/heparan sulfate may be a novel target to control dyslipidemia., (Copyright © 2021 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

Background: Dyslipidemia is an important risk factor in CKD. The liver clears triglyceride-rich lipoproteins (TRL) via LDL receptor (LDLR), LDLR-related protein-1 (LRP-1), and heparan sulfate proteoglycans (HSPGs), mostly syndecan-1. HSPGs also facilitate LDLR degradation by proprotein convertase subtilisin/kexin type 9 (PCSK9). Progressive renal failure affects the structure and activity of hepatic lipoprotein receptors, PCSK9, and plasma cholesterol., Methods: Uninephrectomy- and aging-induced CKD in normotensive Wistar rats and hypertensive Munich-Wistar-Frömter (MWF) rats., Results: Compared with 22-week-old sex- and strain-matched rats, 48-week-old uninephrectomized Wistar-CKD and MWF-CKD rats showed proteinuria, increased plasma creatinine, and hypercholesterolemia (all P <0.05), which were most apparent in hypertensive MWF-CKD rats. Hepatic PCSK9 expression increased in both CKD groups ( P <0.05), with unusual sinusoidal localization, which was not seen in 22-week-old rats. Heparan sulfate (HS) disaccharide analysis, staining with anti-HS mAbs, and mRNA expression of HS polymerase exostosin-1 ( Ext-1 ), revealed elongated HS chains in both CKD groups. Solid-phase competition assays showed that the PCSK9 interaction with heparin-albumin (HS-proteoglycan analogue) was critically dependent on polysaccharide chain length. VLDL binding to HS from CKD livers was reduced ( P <0.05). Proteinuria and plasma creatinine strongly associated with plasma cholesterol, PCSK9, and HS changes., Conclusions: Progressive CKD induces hepatic HS elongation, leading to increased interaction with PCSK9. This might reduce hepatic lipoprotein uptake and thereby induce dyslipidemia in CKD. Therefore, PCSK9/HS may be a novel target to control dyslipidemia., (Copyright © 2021 by the American Society of Nephrology.)

An influential model of the neural mechanisms of creative thought suggests that creativity is manifested in the joint contributions of the Default Mode Network (DMN; a set of regions in the medial PFC, lateral and medial parietal cortex, and the medial temporal lobes) and the executive networks within the dorsolateral PFC. Several empirical reports have offered support for this model by showing that complex interactions between these brain systems account for individual differences in creative performance. The present study examined whether the engagement of these regions in idea generation is modulated by one's eminence in a creativity-related field. Twenty (n ​= ​20) healthy eminent creators from diverse fields of expertise and a 'smart' comparison group of sixteen (n ​= ​16) age- and education-matched non-eminent thinkers were administered a creative generation task (an adaptation of the Alternative Uses Task) and a control perceptual task, while undergoing functional magnetic resonance imaging (fMRI). The participants' verbal responses were recorded through a noise-canceling microphone and were later coded for fluency and accuracy. Behavioral and fMRI analyses revealed commonalities between groups, but also distinct patterns of activation in default mode and executive brain regions between the eminent and the non-eminent participants during creative thinking. We interpret these findings in the context of the well-documented contributions of these regions in the generation of creative ideas as modulated, in this study, by participants' creative eminence., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Endothelial dysfunction (ED) contributes to the high incidence of cardiovascular events in patients undergoing hemodialysis. Syndecan-1 in the endothelial glycocalyx can be shed into the circulation, serving as a biomarker for ED. As Na + is a trigger for glycocalyx shedding, we now tested whether hemodialysis, with higher dialysate Na + concentrations, is associated with more syndecan-1 shedding compared with standard hemodialysis (SHD). In this crossover study in 29 patients, plasma syndecan-1 was repeatedly measured during SHD and during Hemocontrol hemodialysis (HHD), which is characterized by initially higher dialysate and plasma Na + levels. Courses of syndecan-1 were compared with linear mixed models. Syndecan-1 shedding was assessed by area under the curve analysis. Plasma Na + increased early after the start of SHD and HHD, with higher values during HHD (30 min: 142.3 vs. 139.9 mM, P < 0.001). Syndecan-1 increased significantly during both conditions, but the percent change was higher (42.9% vs. 19.5%) and occurred earlier (120 vs. 180 min) during HHD. Syndecan-1 levels were significantly higher at 120 min during HHD compared with SHD ( P < 0.05). Overall, syndecan-1 shedding was higher during HHD compared with SHD (means: 40.4 vs. 19.0 arbitrary units, P = 0.06). Lower predialysis plasma Na + and osmolality were associated with greater intradialytic increases in syndecan-1 levels (both groups, P = 0.001). The rise in plasma syndecan-1 levels was more pronounced and occurred earlier during hemodialysis with higher plasma Na + levels. Although we cannot prove that the rise in plasma syndecan-1 originates from the endothelial glycocalyx, our findings are compatible with Na + -driven endothelial glycocalyx-derived syndecan-1 shedding.

It is well-known that heparin and other glycosaminoglycans (GAGs) inhibit complement activation. It is however not known whether fractionation and/or modification of GAGs might deliver pathway-specific inhibition of the complement system. Therefore, we evaluated a library of GAGs and their derivatives for their functional pathway specific complement inhibition, including the MASP-specific C4 deposition assay. Interaction of human MASP-2 with heparan sulfate/heparin was evaluated by surface plasmon resonance, ELISA and in renal tissue. In vitro pathway-specific complement assays showed that highly sulfated GAGs inhibited all three pathways of complement. Small heparin- and heparan sulfate-derived oligosaccharides were selective inhibitors of the lectin pathway (LP). These small oligosaccharides showed identical inhibition of the ficolin-3 mediated LP activation, failed to inhibit the binding of MBL to mannan, but inhibited C4 cleavage by MASPs. Hexa- and pentasulfated tetrasaccharides represent the smallest MASP inhibitors both in the functional LP assay as well in the MASP-mediated C4 assay. Surface plasmon resonance showed MASP-2 binding with heparin and heparan sulfate, revealing high Kon and Koff rates resulted in a Kd of ~2 μM and confirmed inhibition by heparin-derived tetrasaccharide. In renal tissue, MASP-2 partially colocalized with agrin and heparan sulfate, but not with activated C3, suggesting docking, storage, and potential inactivation of MASP-2 by heparan sulfate in basement membranes. Our data show that highly sulfated GAGs mediated inhibition of all three complement pathways, whereas short heparin- and heparan sulfate-derived oligosaccharides selectively blocked the lectin pathway via MASP-2 inhibition. Binding of MASP-2 to immobilized heparan sulfate/heparin and partial co-localization of agrin/heparan sulfate with MASP, but not C3b, might suggest that in vivo heparan sulfate proteoglycans act as a docking platform for MASP-2 and possibly prevent the lectin pathway from activation., (Copyright © 2020 Talsma, Poppelaars, Dam, Meter-Arkema, Vivès, Gál, Boons, Chopra, Naggi, Seelen, Berger, Daha, Stegeman, van den Born and the COMBAT Consortium.)

The development of ICU-acquired sodium disturbances is not fully understood. Alterations in non-osmotic skin sodium storage, hypothetically inflammation-driven, could play a role. To investigate this in critically ill patients we conducted a patient-control study with skin punch biopsies in patients with sepsis (n = 15), after coronary artery bypass grafting (CABG, n = 15) and undergoing total hip arthroplasty (THA-controls, n = 15) respectively, together representing a range in severity of systemic inflammation. Biopsies were taken within 24 hours (sepsis) and within 2 hours (CABG) after ICU-admission, and prior to arthroplasty. Biopsies were analysed for sodium content. In addition immunostainings and quantitative real time PCR were performed. The primary aim of this study was to detect possible differences in amounts of cutaneous sodium. The secondary aims were to quantify inflammation and lymphangiogenesis with concomitant markers. The highest amounts of both water and sodium were found in patients with sepsis, with slightly lower values after CABG and the lowest amounts in THA-controls. Correlation between water and sodium was 0.5 (p<0.01). In skin biopsies in all groups comparable amounts of macrophages, T-cells and lymph vessels were found. In all groups comparable expression of inflammation markers were found. However, higher mRNA transcript expression levels of markers of lymphangiogenesis were found in patients with sepsis and after CABG. The conjoint accumulation of water and sodium points towards oedema formation. However, the correlation coefficient of 0.5 leaves room for alternative explanations, including non-osmotic sodium storage. No signs of dermal inflammation were found, but upregulation of markers of lymphangiogenesis could indicate future lymphangiogenesis., Competing Interests: The authors have declared that no competing interests exist.

Activated proximal tubular epithelial cells (PTECs) play a crucial role in progressive tubulo-interstitial fibrosis in native and transplanted kidneys. Targeting PTECs by non-viral delivery vectors might be useful to influence the expression of important genes and/or proteins in order to slow down renal function loss. However, no clinical therapies that specifically target PTECs are available at present. We earlier showed that a cationic cell penetrating peptide isolated from South American rattlesnake venom, named crotamine, recognizes cell surface heparan sulfate proteoglycans and accumulates in cells. In healthy mice, crotamine accumulates mainly in kidneys after intraperitoneal (ip) injection. Herein we demonstrate for the first time, the overall safety of acute or long-term treatment with daily ip administrated crotamine for kidneys functions. Accumulation of ip injected crotamine in the kidney brush border zone of PTECs, and its presence inside these cells were observed. In addition, significant lower in vitro crotamine binding, uptake and reporter gene transport and expression could be observed in syndecan-1 deficient HK-2 PTECs compared to wild-type cells, indicating that the absence of syndecan-1 impairs crotamine uptake into PTECs. Taken together, our present data show the safety of in vivo long-term treatment with crotamine, and its preferential uptake into PTECs, which are especially rich in HSPGs such as syndecan-1. In addition to the demonstrated in vitro gene delivery mediated by crotamine in HK-2 cells, the potential applicability of crotamine as prototypic non-viral (gene) delivery nanocarrier to modulate PTEC gene and/or protein expression was confirmed.

Syndecan-1, a transmembrane heparan sulfate proteoglycan, associates with renal and cardiovascular functioning. We earlier reported syndecan-1 to be involved in renal tubular regeneration. We now examined plasma values of syndecan-1 in a hemodialysis cohort and its association with volume and inflammatory and endothelial markers in addition to outcome. Eighty-four prevalent hemodialysis patients were evaluated for their plasma syndecan-1 levels by ELISA before the start of hemodialysis, as well as 60, 180, and 240 min after start of dialysis. Patients were divided into sex-stratified tertiles based on predialysis plasma syndecan-1 levels. We studied the association between plasma levels of syndecan-1 and volume, inflammation, and endothelial markers and its association with cardiovascular events and all-cause mortality using Kaplan-Meier curves and Cox regression analyses with adjustments for gender, age, diabetes, and dialysis vintage. Predialysis syndecan-1 levels were twofold higher in men compared with women ( P = 0.0003). Patients in the highest predialysis plasma syndecan-1 tertile had a significantly higher ultrafiltration rate ( P = 0.034) and lower plasma values of BNP ( P = 0.019), pro-ANP ( P = 0.024), and endothelin ( P < 0.0001) compared with the two lower predialysis syndecan-1 tertiles. No significant associations with inflammatory markers were found. Cox regression analysis showed that patients in the highest syndecan-1 tertile had significantly less cardiovascular events and better survival compared with the lowest syndecan-1 tertile ( P = 0.02 and P = 0.005, respectively). In hemodialysis patients, higher plasma syndecan-1 levels were associated with lower concentrations of BNP, pro-ANP, and endothelin and with better patient survival. This may suggest that control of volume status in hemodialysis patients allows an adaptive tissue regenerative response as reflected by higher plasma syndecan-1 levels.

concepts play a central role in human behaviour and constitute a critical component of the human conceptual system. Here, we investigate the neural basis of four types of abstract concepts, examining their similarities and differences through neuroimaging meta-analyses. We examine numerical and emotional concepts, and two higher-order abstract processes, morality judgements and theory of mind. Three main findings emerge. First, representation of abstract concepts is more widespread than is often assumed. Second, representations of different types of abstract concepts differ in important respects. Each of the domains examined here was associated with some unique areas. Third, some areas were commonly activated across domains and included inferior parietal, posterior cingulate and medial prefrontal cortex. We interpret these regions in terms of their role in episodic recall, event representation and social-emotional processing. We suggest that different types of abstract concepts can be represented and grounded through differing contributions from event-based, interoceptive, introspective and sensory-motor representations. The results underscore the richness and diversity of abstract concepts, argue against single-mechanism accounts for representation of all types of abstract concepts and suggest mechanisms for their direct and indirect grounding.This article is part of the theme issue 'Varieties of abstract concepts: development, use and representation in the brain'., (© 2018 The Author(s).)

Proteinuria is an important cause of tubulointerstitial damage. Anti-proteinuric interventions are not always successful, and residual proteinuria often leads to renal failure. This indicates the need for additional treatment modalities by targeting the harmful downstream consequences of proteinuria. We previously showed that proteinuria triggers renal lymphangiogenesis before the onset of interstitial inflammation and fibrosis. However, the interrelationship of these interstitial events in proteinuria is not yet clear. To this end, we specifically blocked lymphangiogenesis (anti-VEGFR3 antibody), monocyte/macrophage influx (clodronate liposomes) or lymphocyte and myofibroblast influx (S1P agonist FTY720) separately in a rat model to investigate the role and the possible interaction of each of these phenomena in tubulointerstitial remodeling in proteinuric nephropathy. Proteinuria was induced in 3-month old male Wistar rats by adriamycin injection. After 6 weeks, when proteinuria has developed, rats were treated for another 6 weeks by anti-VEGFR3 antibody, clodronate liposomes or FTY720 up to week 12. In proteinuric rats, lymphangiogenesis, influx of macrophages, T cells and myofibroblasts, and collagen III deposition and interstitial fibrosis significantly increased at week 12 vs week 6. Anti-VEGFR3 antibody prevented lymphangiogenesis in proteinuric rats, however, without significant effects on inflammatory and fibrotic markers or proteinuria. Clodronate liposomes inhibited macrophage influx and partly reduced myofibroblast expression; however, neither significantly prevented the development of lymphangiogenesis, nor fibrotic markers and proteinuria. FTY720 prevented myofibroblast accumulation, T-cell influx and interstitial fibrosis, and partially reduced macrophage number and proteinuria; however, it did not significantly influence lymphangiogenesis and collagen III deposition. This study showed that proteinuria-induced interstitial fibrosis cannot be halted by blocking lymphangiogenesis or the influx of macrophages. On the other hand, FTY720 treatment did prevent T-cell influx, myofibroblast accumulation and interstitial fibrosis, but not renal lymphangiogenesis and proteinuria. We conclude that tubulointerstitial fibrosis and inflammation are separate from lymphangiogenesis, at least under proteinuric conditions., (© 2015. Published by The Company of Biologists Ltd.)

Syndecan-1 is a transmembrane heparan sulfate proteoglycan involved in regenerative growth and cellular adhesion. We hypothesized that the induction of tubular syndecan-1 is a repair response to incipient renal damage in apparently stable, uncomplicated renal transplant recipients. We quantified tubular syndecan-1 in unselected renal protocol biopsies taken 1 yr after transplantation. Spearman rank correlation analysis revealed an inverse correlation between tubular syndecan-1 expression and creatinine clearance at the time of biopsy (r = -0.483, P < 0.03). In a larger panel of protocol and indication biopsies from renal transplant recipients, tubular syndecan-1 correlated with tubular proliferation marker Ki67 (r = 0.518, P < 0.0001). In a rat renal transplantation model, 2 mo after transplantation, mRNA expression of syndecan-1 and its major sheddase, A disintegrin and metalloproteinase-17, were upregulated (both P < 0.03). Since shed syndecan-1 might end up in the circulation, in a stable cross-sectional human renal transplant population (n = 510), we measured plasma syndecan-1. By multivariate regression analysis, we showed robust independent associations of plasma syndecan-1 with renal (plasma creatinine and plasma urea) and endothelial function parameters (plasma VEGF-A, all P < 0.01). By various approaches, we were not able to localize syndecan-1 in vessel wall or endothelial cells, which makes shedding of syndecan-1 from the endothelial glycocalyx unlikely. Our data suggest that early damage in transplanted kidneys induces repair mechanisms within the graft, namely, tubular syndecan-1 expression for tubular regeneration and VEGF production for endothelial repair. Elevated plasma syndecan-1 levels in renal transplantation patients might be interpreted as repair/survival factor related to loss of tubular and endothelial function in transplanted kidneys., (Copyright © 2015 the American Physiological Society.)

The role of the two hemispheres in processing metaphoric language is controversial. While some studies have reported a special role of the right hemisphere (RH) in processing metaphors, others indicate no difference in laterality relative to literal language. Some studies have found a role of the RH for novel/unfamiliar metaphors, but not conventional/familiar metaphors. It is not clear, however, whether the role of the RH is specific to metaphor novelty, or whether it reflects processing, reinterpretation or reanalysis of novel/unfamiliar language in general. Here we used functional magnetic resonance imaging (fMRI) to examine the effects of familiarity in both metaphoric and non-metaphoric sentences. A left lateralized network containing the middle and inferior frontal gyri, posterior temporal regions in the left hemisphere (LH), and inferior frontal regions in the RH, was engaged across both metaphoric and non-metaphoric sentences; engagement of this network decreased as familiarity decreased. No region was engaged selectively for greater metaphoric unfamiliarity. An analysis of laterality, however, showed that the contribution of the RH relative to that of LH does increase in a metaphor-specific manner as familiarity decreases. These results show that RH regions, taken by themselves, including commonly reported regions such as the right inferior frontal gyrus (IFG), are responsive to increased cognitive demands of processing unfamiliar stimuli, rather than being metaphor-selective. The division of labor between the two hemispheres, however, does shift towards the right for metaphoric processing. The shift results not because the RH contributes more to metaphoric processing. Rather, relative to its contribution for processing literals, the LH contributes less.

Syndecan-1 is a transmembrane heparan sulfate (HS) proteoglycan present on hepatocytes and involved in uptake of triglyceride-rich lipoproteins via its HS polysaccharide side chains. We hypothesized that altered hepatic syndecan-1 metabolism could be involved in dyslipidemia related to renal transplantation. In a rat renal transplantation model elevated plasma triglycerides were associated with fivefold increased expression of hepatic syndecan-1 mRNA (p < 0.01), but not protein. Expression of syndecan-1 sheddases (ADAM17, MMP9) and heparanase was significantly up-regulated after renal transplantation (all p < 0.05). Profiling of HS side chains revealed loss of hepatic HS upon renal transplantation accompanied by significant decreased functional capacity for VLDL binding (p = 0.02). In a human renal transplantation cohort (n = 510), plasma levels of shed syndecan-1 were measured. Multivariate analysis showed plasma syndecan-1 to be independently associated with triglycerides (p < 0.0001) and inversely with HDL cholesterol (p < 0.0001). Last, we show a physical association of syndecan-1 to HDL from renal transplant recipients (RTRs), but not to HDL from healthy controls. Our data suggest that after renal transplantation loss of hepatic HS together with increased syndecan-1 shedding hampers lipoprotein binding and uptake by the liver contributing to dyslipidemia. Our data open perspectives toward improvement of lipid profiles by targeted inhibition of syndecan-1 catabolism in renal transplantation., (© Copyright 2014 The American Society of Transplantation and the American Society of Transplant Surgeons.)

We address the question of what physical resources are required and sufficient to store classical information. While there is no lower bound on the required energy or space to store information, we find that there is a nonzero lower bound for the product P = of these two resources. Specifically, we prove that any physical system of mass m and d degrees of freedom that stores S bits of information will have a lower bound on the product P that is proportional to d2/m(exp(S/d) - 1)2. This result is obtained in a nonrelativistic, quantum mechanical setting, and it is independent of earlier thermodynamical results such as the Bekenstein bound on the entropy of black holes.

Objective: To increase equitable access to life insurance for HIV-positive individuals by identifying subgroups with lower relative mortality., Design: Collaborative analysis of cohort studies., Methods: We estimated relative mortality from 6 months after starting antiretroviral therapy (ART), compared with the insured population in each country, among adult patients from European cohorts participating in the ART Cohort Collaboration (ART-CC) who were not infected via injection drug use, had not tested positive for hepatitis C, and started triple ART between 1996-2008. We used Poisson models for mortality, with the expected number of deaths according to age, sex and country specified as offset., Results: There were 1236 deaths recorded among 34,680 patients followed for 174,906 person-years. Relative mortality was lower in patients with higher CD4 cell count and lower HIV-1 RNA 6 months after starting ART, without prior AIDS, who were older, and who started ART after 2000. Compared with insured HIV-negative lives, estimated relative mortality of patients aged 20-39 from France, Italy, United Kingdom, Spain and Switzerland, who started ART after 2000 had 6-month CD4 cell count at least 350 cells/μl and HIV-1 RNA less than 10⁴ copies/ml and without prior AIDS was 459%. The proportion of exposure time with relative mortality below 300, 400, 500 and 600% was 28, 43, 61 and 64%, respectively, suggesting that more than 50% of patients (those with lower relative mortality) could be insurable., Conclusion: The continuing long-term effectiveness of ART implies that life insurance with sufficiently long duration to cover a mortgage is feasible for many HIV-positive people successfully treated with ART for more than 6 months., (© 2013 Creative Common License)

Introduction: The Coasys® Plus C (Behnk Elektronik, distributed by Roche Diagnostics) is a coagulation analyzer for small to midsize clinical chemistry laboratories. We performed a laboratory evaluation., Materials and Methods: After a familiarization period the dead volume, carry-over, capacity, within-assay reproducibility and imprecision were determined for the tests aPTT (STA APTT en STA Cephascreen), PT (STA Neoplastin Plus, STA Neoplastin R), INR (Hepato Quick), fibrinogen (STA Fibrinogen), antithrombin (Antithrombin III) and D-Dimer (Tina-quant D-Dimer Gen.2). A precision test and patient comparison with a STA-R Evolution® system were performed for aPTT (STA Cephascreen), PT (STA Neoplastin R), fibrinogen, antithrombin and D-Dimer., Results: No carry-over was detected. The analyzer performed on average 66 measurements per hour. Within-assay reproducibility (% with normal/abnormal/extreme result): STA APTT 2.4/1.7/2.7; aPTT Cephascreen 1.4/3.1/1.8; PT Neoplastin Plus 1.2/2.1/1.7; PT Neoplastin R 2.3/-/3.1; INR Hepato Quick 0.6/0.9/1.9; fibrinogen 4.8/3.0/4.5; antithrombin 1.3/4.1/3.8; D-Dimer 19.9/4.3/4.3. Total imprecision (% with control 1/control 2/human pooled plasma): STA APTT 3.1/1.4/2.1; APTT Cephascreen 2.3/2.6/2.0; PT Neoplastin Plus 3.3/1.3/4.1; PT Neoplastin R 3.7/4.0/3.5; INR Hepato Quick 1.5/4.3/1.4; fibrinogen 3.4/5.8/8.1; antithrombin 3.0/6.4/2.4; D-Dimer 4.6/2.2/30.3. The correlation between the Coasys® Plus C and the STA-R Evolution® was good for aPTT, PT, antithrombin and fibrinogen. Some differences were observed for extreme deviant results for fibrinogen. For the analysis of D-Dimer, no sufficient correlation was found between the two analyzers., Conclusions: The Coasys® Plus C analyzer is fast, easy to handle and safe for the personnel. Its analytical performance makes it suitable for use in a clinical chemistry laboratory., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Objective: To determine the effect of topically applied amelogenin extracellular matrix protein(AEMP) in patients with non-healing venous leg ulcers combined with atrophie blanche., Method: This retrospective case series of patients with non-healing venous leg ulcers with atrophie blanche of the distal proportion of their lower legs, where non-healing was defined as no progress toward healing for 3 months previously, under standard therapy. Patient records were reviewed for associated diseases, wound diagnoses, distal blood pressure, previous treatments and changes in wound area. Patients were treated with AEMP once a week, for a period of 12 weeks, or until full healing., Results: Eleven patient records were reviewed retrospectively. The median age of the patients was 81 years (range 40-95 years), with a mean wound size of 4.7 ± 3.Scm2 and median wound duration of 6 months (range 3-444 months).AII patients had venous or combined arterial/venous insufficiency. After 12 weeks' treatment with AEMP, complete healing, defined as I 00% re-epithelialisation, was documented in four patients (36%), marked improvement(> SO% epithelialisation) in three patients (54%, 55% and 83% wound closure, respectively), slight improvement in one patient (9.4% wound closure), no change for two patients and worsening in one.AEMP was well tolerated, and no patients reported side effects., Conclusion: The results of this retrospective study suggest that AEMP improves healing in chronic venous leg ulcers combined with atrophie blanche.

Application of human adenovirus type 5 (Ad5) derived vectors for cancer gene therapy has been limited by the poor cell surface expression, on some tumor cell types, of the primary Ad5 receptor, the coxsackie-adenovirus-receptor (CAR), as well as the accumulation of Ad5 in the liver following interaction with blood coagulation factor X (FX) and subsequent tethering of the FX-Ad5 complex to heparan sulfate proteoglycan (HSPG) on liver cells. As an alternative vector, mouse adenovirus type 1 (MAV-1) is particularly attractive, since this non-human adenovirus displays pronounced endothelial cell tropism and does not use CAR as a cellular attachment receptor. We here demonstrate that MAV-1 uses cell surface heparan sulfate proteoglycans (HSPGs) as primary cellular attachment receptor. Direct binding of MAV-1 to heparan sulfate-coated plates proved to be markedly more efficient compared to that of Ad5. Experiments with modified heparins revealed that the interaction of MAV-1 to HSPGs depends on their N-sulfation and, to a lesser extent, 6-O-sulfation rate. Whereas the interaction between Ad5 and HSPGs was enhanced by FX, this was not the case for MAV-1. A slot blot assay demonstrated the ability of MAV-1 to directly interact with FX, although the amount of FX complexed to MAV-1 was much lower than observed for Ad5. Analysis of the binding of MAV-1 and Ad5 to the NCI-60 panel of different human tumor cell lines revealed the preference of MAV-1 for ovarian carcinoma cells. Together, the data presented here enlarge our insight into the HSPG receptor usage of MAV-1 and support the development of an MAV-1-derived gene vector for human cancer therapy.

The acquisition of bidirectional action-effect associations plays a central role in the ability to intentionally control actions. Humans learn about actions not only through active experience, but also through observing the actions of others. In Experiment 1, we examined whether action-effect associations can be acquired by observational learning. To this end, participants observed how a model repeatedly pressed two buttons during an observation phase. Each of the buttonpresses led to a specific tone (action effect). In a subsequent test phase, the tones served as target stimuli to which the participants had to respond with buttonpresses. Reaction times were shorter if the stimulus-response mapping in the test phase was compatible with the action-effect association in the observation phase. Experiment 2 excluded the possibility that the impact of perceived action effects on own actions was driven merely by an association of spatial features with the particular tones. Furthermore, we demonstrated that the presence of an agent is necessary to acquire novel action-effect associations through observation. Altogether, the study provides evidence for the claim that bidirectional action-effect associations can be acquired by observational learning. Our findings are discussed in the context of the idea that the acquisition of action-effect associations through observation is an important cognitive mechanism subserving the human ability for social learning.

Embodied theories of language processing suggest that this motor simulation is an automatic and necessary component of meaning representation. If this is the case, then language and action systems should be mutually dependent (i.e., motor activity should selectively modulate processing of words with an action-semantic component). In this paper, we investigate in two experiments whether evidence for mutual dependence can be found using a motor priming paradigm. Specifically, participants performed either an intentional or a passive motor task while processing words denoting manipulable and nonmanipulable objects. The performance rates (Experiment 1) and response latencies (Experiment 2) in a lexical-decision task reveal that participants performing an intentional action were positively affected in the processing of words denoting manipulable objects as compared to nonmanipulable objects. This was not the case if participants performed a secondary passive motor action (Experiment 1) or did not perform a secondary motor task (Experiment 2). The results go beyond previous research showing that language processes involve motor systems to demonstrate that the execution of motor actions has a selective effect on the semantic processing of words. We suggest that intentional actions activate specific parts of the neural motor system, which are also engaged for lexical-semantic processing of action-related words and discuss the beneficial versus inhibitory nature of this relationship. The results provide new insights into the embodiment of language and the bidirectionality of effects between language and action processing.

We study how much noise can be tolerated by a universal gate set before it loses its quantum-computational power. Specifically we look at circuits with perfect stabilizer operations in addition to imperfect nonstabilizer gates. We prove that for all unitary single-qubit gates there exists a tight depolarizing noise threshold that determines whether the gate enables universal quantum computation or if the gate can be simulated by a mixture of Clifford gates. This exact threshold is determined by the Clifford polytope spanned by the 24 single-qubit Clifford gates. The result is in contrast to the situation wherein nonstabilizer qubit states are used; the thresholds in that case are not currently known to be tight.

We address the problem of estimating the phase phi given N copies of the phase-rotation gate uphi. We consider, for the first time, the optimization of the general case where the circuit consists of an arbitrary input state, followed by any arrangement of the N phase rotations interspersed with arbitrary quantum operations, and ending with a general measurement. Using the polynomial method, we show that, in all cases where the measure of quality of the estimate phi for phi depends only on the difference phi-phi, the optimal scheme has a very simple fixed form. This implies that an optimal general phase estimation procedure can be found by just optimizing the amplitudes of the initial state.

Anti-angiogenic therapy is emerging as a valuable tool in the treatment of patients with cancer. As VEGF is a central target in anti-angiogenic therapy, its levels in the circulation might be relevant in selecting tumor types or patients likely to respond to this treatment. Additional VEGF has been recognized as a key factor in the pathogenesis of diabetic retinopathy. Recently anti-angiogenic therapy has been advocated in this situation.We measured VEGF levels in whole blood in 42 patients with high grade (n = 26) and low grade (n = 16) end stage cancer, and in 28 healthy controls and 37 patients with diabetes related vascular disease. Only 2/26 patients in the group of high grade cancer had significantly elevated VEGF levels, 1/16 in the low grade group and 1/28 in the healthy control group. In contrast, in 10/37 diabetic patients the mean VEGF levels were significantly elevated compared to the other groups. The mean level in these diabetic patients was significantly elevated compared to the other groups.These data indicate the limitation of the use of circulating VEGF levels as a potential selection criterion for anti-angiogenic therapy in cancer patients and suggest further studies into its application in the management of diabetic complications.

A female infant with dysmorphic facial features, psychomotor retardation, and clitoris hypertrophy is described. Molecular cytogenetic analyses revealed a de novo unbalanced translocation, causing partial monosomy 1p36 and partial trisomy 18q22. Monosomy 1p was confirmed by FISH, and trisomy of the distal part of chromosome 18q was demonstrated by microFISH. Gene copy number changes in these chromosomal regions were determined by array-CGH. The absence of a number of facial dysmorphic signs, and the presence of clitoris hypertrophy indicate that the combination of a del(1p36->pter) with a dup(18q22->qter) may lead to a unique phenotypic constellation. The findings at birth and at age 12 years in our patient are compared with genotype-phenotype correlations discussed in the literature.

Our study focused on the influence of herpes simplex virus thymidine kinase (HSV-tk) expression and ganciclovir (GCV) treatment on the sensitivity of C6 glioma cells to frequently used chemotherapeutic drugs, i.e. adriamycin (ADR), cisplatin (CDDP), 5-fluorouracil (5-FU), and methotrexate (MTX). Transfection with HSV-tk revealed an increased sensitivity to GCV and CDDP and a decreased sensitivity to ADR and MTX. No significant differences were found in sensitivity to 5-FU. Combined treatment in a HSV-tk negative cell line revealed an additive effect when GCV was combined with ADR, whereas an antagonistic effect was found when GCV was combined with CDDP, 5-FU, or MTX. Comparable results were obtained in an HSV-tk positive cell line, apart from CDDP, which showed an additive effect. In conclusion, both HSV-tk transfection and subsequent GCV treatment can influence the sensitivity of tumor cells to various chemotherapeutic drugs in an antagonistic manner. Therefore, combining HSV-tk/GCV gene therapy with chemotherapy might not always be beneficial.

We report a girl with severe retardation of expressive speech development carrying a small, supernumerary ring chromosome derived from the proximal region of the long arm of chromosome 7. The r(7) chromosome is present in 50% of lymphocytes. We also review the six additional cases with a supernumerary r(7) chromosome reported in the literature. Among these patients, a severe retardation of productive language capabilities is seen as a shared clinical feature, irrespective of the degree of mosaicism as detected in blood. The dysmorphisms in these patients are minor and no shared congenital abnormalities seen. We, therefore, recommend chromosomal investigations in children with unexplained, disproportionately retarded expressive speech performance. Because speech and language acquisition are subject to genetic influences, we investigated whether there are genes on the r(7) chromosome that may affect brain development or function in a dosage-dependent manner. We found that both in our patient and in four patients described by others, the supernumerary r(7) chromosome contains the region from the centromere up to marker D7S613 located at 7q11.23. We speculate that the effects on speech acquisition are mediated by the supernumerary copies of the STX1A and LIMK1 genes, which are both located in this region and known to suppress neurite growth when overexpressed in vitro., ((c) 2004 Wiley-Liss, Inc.)

Objective: This study aimed to identify a marker chromosome and characterize the short arm of a derivative chromosome 5 in a foetus with the following karyotype: mos 47,XX,del(5)(p?),+i(5)(p10)[50]/48,XX,del(5)(p?),+i(5)(p10),+mar[25]., Method: Amniocentesis was performed in the 26th week of pregnancy because of ultrasound abnormalities (polyhydramnion and decreased amount of gastric filling). All classic banding techniques were performed. FISH and microdissection combined with reverse painting were used to reveal the exact origin of the marker and any extra material on the deleted chromosome 5p. The parents decided to continue the pregnancy and we compared the clinical features of the child born in week 34 with data from the literature on trisomy 5p. The possible contribution of trisomy of the centromeric region of chromosome 8 and trisomy 8p23.3-->8pter to this clinical picture was evaluated., Results: GTG banding showed one normal and two aberrant chromosomes 5 [del(5)(p?) and i(5)(p10)] in all the cells examined. Furthermore, a supernumerary marker chromosome was present in approximately 30% of the cells. The marker was CBG positive and positive with the pancentromere probe, but dystamicinA/DAPI negative. It did not contain NOR-positive satellites. FISH proved this marker to be derived from the centromeric region of chromosome 8. MicroFISH disclosed the aberrant chromosome 5 as der(5)t(5;8)(p10;p23.3). The parent's karyotypes were normal. The baby showed the characteristic features of trisomy 5p syndrome. She died at the age of 15 days after cardiorespiratory arrest., Conclusion: The karyotype was interpreted as mos 47,XX,add(5)(p10).rev ish der(5)t(5;8)(p10;p23.3),+i(5)(p10) (WCP5+,D5S23+)[50]/48,XX,add(5)(p10).rev ish der(5)t(5;8)(p10;p23.3),+i(5)(p10)(WCP5+,D5S23+),+mar.ish 8(p10q10)(D8Z2+,WCP8-)[25]. Therefore, the baby had complete trisomy 5p, with trisomy of the distal part of 8p and of the centromeric region of chromosome 8. The clinical significance of de novo marker chromosomes is a major problem in prenatal counselling. Molecular cytogenetic tools such as FISH and microFISH are indispensable for characterizing markers and determining the breakpoints more precisely in deleted chromosomes., (Copyright 2003 John Wiley & Sons, Ltd.)

Background: Cornea-derived transcript 6 (CDT6, also known as AngX) has been described to inhibit tumour growth in a human melanoma growing in nude mice. In another report, however, enhancement of tumour growth in comparable circumstances occurred. We therefore studied the effect of the same gene in different circumstances, using an immune competent mouse model., Materials and Methods: In this report we describe the generation of a stably CDT6-expressing clone of the murine melanoma cell line B16-F10. These cells were implanted in female C57/B16 mice with empty vector transfected cells as control., Results: We found no significant inhibition or stimulation of either lag-time or doubling-time of the tumours. In addition no effect of the CDT6 gene product was found on proliferation of endothelial cells in vitro., Conclusion: In contrast to an immune deficient mouse model, no anti-tumour effect could be detected with the CDT6 gene product in an immune competent mouse model.

We report the realization of a nuclear magnetic resonance computer with three quantum bits that simulates an adiabatic quantum optimization algorithm. Adiabatic quantum algorithms offer new insight into how quantum resources can be used to solve hard problems. This experiment uses a particularly well-suited three quantum bit molecule and was made possible by introducing a technique that encodes general instances of the given optimization problem into an easily applicable Hamiltonian. Our results indicate an optimal run time of the adiabatic algorithm that agrees well with the prediction of a simple decoherence model.

Background: Cornea-derived transcript 6 (CDT6 = AngX) has been shown to have an anti-tumor effect., Materials and Methods: We transfected the murine melanoma cell line B16-F10 with the CDT6 gene and compared the sensitivity to cytostatic drugs of the resulting cell line, B16-CDT6, to that of the empty vector-transfected control cell line B16-CMV., Results: The B16-CDT6 line showed a significantly increased sensitivity to doxorubicin as well as a tendency towards a decreased sensitivity to cisplatin. To further resolve the mechanism of the increase in sensitivity to doxorubicin, we also tested the cytotoxic agents vincristine, etoposide and taxol. However no difference in sensitivity for these drugs was found between the B16-CDT6 and the control cell line., Conclusion: CDT6 increased the sensitivity to doxorubicin in a mechanism probably not related to interference with the efflux pumps MRP, Pgp or the DNA-associated target enzyme topoisomerase II. Altered gene expression induced by gene therapy might influence tumor sensitivity to chemotherapy.

We describe a family with an insertion 12;9 translocation occurring in a balanced form in a mother and two sons, but in an unbalanced form in the proband, resulting in trisomy of chromosome region 9p22-->9p24. The proband manifests typical features of trisomy 9p; the clinical signs were mental and growth retardation, microcephaly, epicanthus, low-set ears, micrognathia, clinodactyly and hypoplastic phalanges of the fifth fingers, hypoplasia or absence of toenails, and extremely small genitals. The GTG-banded findings were confirmed using (micro)FISH. Intriguingly, the mother and the two carrier sons exhibited major learning difficulties that were not present in the non-carrier sister of the mother: this may be due to a gene disruption or induction of abnormal expression. Dysmorphic features were not present in the three carriers. We compare our clinical and cytogenetic findings with other cases of partial trisomy 9p reported in the literature.

Magnesium ions were detected in sodium citrate solutions in several lots of evacuated blood collection tubes. The mean concentrations ranged between 1.3 and 1.6 mmol/L. Magnesium was also present in the rubber stoppers of the blood collection tubes and could be leached into the citrate solution. It was shown that magnesium added to citrated plasma shortened the prothrombin time of both coumarin and normal plasma. The effect of magnesium was relatively greater on coumarin than on normal plasma resulting in reduced prothrombin time ratio. Shortening of the prothrombin time was also observed when magnesium chloride was added to dialysed plasma, i.e., in the absence of citrate. These results indicate that magnesium contamination can interfere with accurate INR determination in the control of oral anticoagulant therapy.

Cisplatin (CDDP) is an extremely active drug in the treatment of germ-cell tumours. Earlier, we found an unexpected inverse correlation between the total amount of sulfhydryl groups and CDDP sensitivity in a panel of 3 human germ-cell-tumour and 3 colon-carcinoma cell lines. Major components of the sulfhydryl groups are glutathione and metallothionein (MT). We further investigated a possible role of MT in the CDDP sensitivity of germ-cell tumours. MT levels and functionality of the germ-cell-tumour and colon-carcinoma cell lines were found to be inversely correlated with CDDP sensitivity. No difference in sub-cellular localization of MT could be observed among the types of cell lines. In agreement with the in vitro data, immunohistochemical detection of MT was high in 11/14 primary human germ-cell tumours and low in 7/7 human colon carcinomas. MT-protein expression in primary germ-cell tumours did not discriminate between responding and non-responding patients. As compared with the primary tumours, MT-protein expression decreased in 5/7 post-chemotherapy residual vital tumours or remained undetectable (2/7). MT-protein expression in the germ-cell tumours was not related to total p53-protein expression. In summary, over-expression of MT was found in germ-cell tumours, both in cell lines and in human tumours. Although MT-protein over-expression seems to be associated with the CDDP sensitivity of germ-cell tumours, MT-protein expression in primary germ-cell tumours did not differ between responding and non-responding patients and therefore cannot be used to predict response to chemotherapy., (Copyright 2000 Wiley-Liss, Inc.)

To detect low-level DNA platination, a sensitive immunocyto- and histochemical technique was developed using a polyclonal antibody. The antibody GPt, derived after immunization of rabbits with highly platinated DNA and purified with affinity chromatography, detected the main platinum (Pt)-containing intrastrand and interstrand adducts. Double-fluorescence microscopy image analysis was used to quantify Pt-DNA adducts with Hoechst 33258 fluorescence to locate the nuclei and with fluorescein isothiocyanate fluorescence to measure the immunosignal. A two- to five-fold dose-dependent difference in the level of cisplatin (CDDP)-induced Pt-DNA adducts between a CDDP-sensitive and -resistant human tumour cell line was detected. Large differences in Pt-DNA adduct levels after in vitro CDDP incubation between human buccal cells, lymphocytes and biopsies of different tumour types were observed. Pt-DNA adduct levels were fivefold higher in human testicular tumours than in colon tumours, representing CDDP-sensitive and -resistant tumours, respectively, in the clinic. These data suggest the possibility of predictive testing by measuring Pt-DNA adduct levels. Pt-DNA adducts in patients after treatment with CDDP were shown in normal buccal cells and in imprints of fresh tumour biopsies as well as in paraffin-embedded tumour cells. The analysis of Pt-DNA adducts at a single-cell level in small samples of normal and tumour cells during and/or after treatment is feasible with GPt and will hopefully enable more selective treatment of patients.

In this study, the mechanism(s) by which heat increases cis-diamminedichloroplatinum (cisplatin, cDDP) sensitivity in cDDP-sensitive and -resistant cell lines of murine as well as human origin were investigated. Heating cells at 43 degrees C during cDDP exposure was found to increase drug accumulation significantly in the cDDP-resistant cell lines but had little effect on drug accumulation in the cDDP-sensitive cell lines. DNA adduct formation, however, was significantly increased in all cell lines studied. Furthermore, ongoing formation of platinum (Pt)-DNA adducts after the end of cDDP treatment was enhanced and/or adduct removal was decreased in heated cells, resulting in relatively more DNA damage remaining at 24 h after the end of cDDP exposure. Correlation plots with survival revealed weak correlations with cellular Pt accumulation (r2 = 0.59) and initial Pt-DNA adduct formation (r2 = 0.64). Strong correlations, however, were found with Pt-DNA adducts at 6 h (r2 = 0.97) and 24 h (r2 = 0.89) after the incubation with the drug. In conclusion, the mechanism by which heat sensitizes cells for cDDP action seems to be the sum of multiple factors, which comprise heat effects on accumulation, adduct formation and adduct processing. This mechanism did not seem to differ between cDDP-sensitive and -resistant cells, emphasizing the potential of hyperthermia to reduce cDDP resistance.

A method is described for the semi-automated measurement of inulin concentrations in serum and urine in the presence of glucose. The concentration of glucose is measured simultaneously and is used to correct for almost all "inulin-like" interferences. The inulin standard curve is linear over a wide range (5-2500 mg/l). Between-run precision is < 6% and recovery from spiked sera is 98%. This method offers the possibility of measuring glomerular filtration rate in patients with varying glucose concentrations (e.g. diabetics).

In 14 ADPKD patients the total body clearance and the urinary clearance of inulin using the constant infusion method were compared with the "single-shot" technique. Triplicate measurements of both clearances by each infusion method were obtained in 12 out of 14 patients. A high correlation was found between the total body clearance and the urinary clearance for both the constant infusion method (r = 0.96) and the single injection technique (r = 0.96). The coefficient of variation for the total body clearance of inulin was significantly lower for the constant infusion method and the single injection technique (7.8% and 7.1%) than for the urinary clearance of inulin (11.3% vs. 9.7%, P < 0.05). A constant overestimation of the urinary clearance by the total body clearance was observed with both methods (constant infusion method 8.3 ml.min-1 x 1.73 m-2 and single injection technique 13.4 ml.min-1 x 1.73 m-2). No concentration-dependent clearance was present. Determination of plasma inulin, especially at low levels, showed substantial interference by glucose. We conclude that, taking into account a constant overestimate of urinary clearance by the total body clearance of inulin, the single injection total body clearance possesses the best reproducibility and shows a good agreement with the conventional urinary clearance, which can be calculated by: GFR = TBCLss-13.1 ml.min-1 x 1.73 m-2 (in the range of 28 to 124 ml.min-1 x 1.73 m-2).

In this paper we describe, as an example, how we obtained the information needed to evaluate a newly introduced protocol for ordering X-rays for ankle trauma patients. Extensive use was made of available data and facilities of the hospital information system (HIS). Procedures for collecting the required additional data, which were not recorded in the HIS but were needed to evaluate the protocol, were embedded in the current medical and administrative routine of the emergency room. These additional data were also stored in the HIS. Periodically all data were downloaded to a personal computer to analyse the impact of using the protocol on quality of care and costs. In total 1241 patients entered the study, and for 1149 patients a complete dataset was obtained. The sensitivity and specificity of the protocol at the threshold value which was used during the initial study period was 0.77 and 0.80. The reduction in the number of ankle X-rays due to the protocol was significant when compared with a strategy of ordering an X-ray for every ankle trauma patient visiting the emergency room.

The influence of X-irradiation on the sensitivity of the rat sciatic nerve to local hyperthermia was investigated. A 10 or 20 mm long segment of the nerve was irradiated intraoperatively using 50 kV X-rays. Hyperthermia (30 min at 45 degrees C), was applied to the irradiated part (over a length of 5 mm) of the nerve using a brass thermode. Functional damage to the nerve was assessed using the toe-spreading test, which mainly assesses the motor function of the sciatic nerve. Radiation alone (doses up to 70 Gy) did not lead to detectable damage for at least 90 weeks. Hyperthermia alone (30 min at 45 degrees C) resulted in complete loss of motor function. This function loss was transient and complete recovery took place in about 4 weeks. Recovery time was scored as the number of days between hyperthermia and the day on which 50% of the motor function had returned. Irradiation (35 Gy) of a nerve segment, which included the heated part, resulted in a delayed recovery from the heat treatment compared to controls (heat only). The time interval and sequence between irradiation and hyperthermia hardly influenced the recovery delay. The size of the irradiated nerve segment did influence the recovery delay. Irradiation of a 20 mm nerve segment led to longer recovery delays than irradiation of a 10 mm segment (a delay of 5-10 days and 1-5 days respectively). A dose-response relation for the irradiation-induced delay in recovery was observed when a large segment (20 mm) of the nerve was irradiated immediately after heat with a dose ranging from 5 to 40 Gy. The delay in heat recovery was dose-dependent below 20 Gy, but after radiation doses above 20 Gy the recovery delay remained almost constant.

During a period of 11 years in specialist practice, the authors notified 309 cases of occupationally-conditioned disease and occupational accidents. The majority of diseases were in the locomotor system. The authors reviewed the sex and age distributions and the distribution of the conditions involved. Their share of notifications from specialist practice during the period 1983-1986 was also reviewed together with the relevant specialties and the action taken by the Workers Supervision Authorities. The authors found their share in the notifications was relatively excessive but that it was declining because a great number of specialists have begun to notify conditions in the locomotor system. In addition, the authors have observed a considerable reduction in the rime taken to analyse cases by the Workers Supervision Authorities and an increase in the number of factory visits. The number of factory visits on account of the authors' notifications constituted 11% which shows that notification of occupationally-conditioned diseases is worth while.

The cell-lineage of Bithynia tentaculata L. has been studied through the formation of the fourth quartet of micromeres, i.e., the 81-cell stage. Up to the 28-cell stage, the cleavage pattern is radially symmetrical, and the divisions take place synchronously in the quadrants. Bilateral symmetry is established by the formation of the mesentoblast 4d at the future dorsal side. A resting stage preceding the formation of 4d is lacking. The mesentoblast 4d is formed earlier than the other fourth quartet cells, and it divides twice before 4a-4c are formed. Bilateral symmetry in the molluscan cross becomes apparent by different divisions of the tipcell and the basal cell in the dorsal arm. Dorsoventrality is progressively corroborated by the divisions in the second and third quartets of micromeres, which take a different course in the A and B quadrants on the one side and in the C and D quadrants on the other side., (Copyright © 1986 Wiley-Liss, Inc.)

The significance of the first quartet of micromeres for the morphogenesis ofBithynia - a polar lobe-forming gastropod-has been studied by deletion experiments. After removal of the whole first quartet at the 8-cell stage a dorsoventrally organized veliger larva is formed. Apparently, an interaction between the animal micromeres and a vegetal macromere, which is essential for the origin of a dorsoventral organization in equally cleaving gastropods, is not required in polar lobe forming eggs. It is concluded that in these eggs dorsoventrality is determined by segregation of the polar lobe. The embryos, in which the first quartet has been removed, never develop head structures. This indicates that the capacity to form head structures is restricted to the first quartet of micromeres. Deletion of a specific first quartet micromere (1a, 1b, 1c or 1d) showed, however, that the individual cells of this quartet are not strictly determined right from their origin. Frequently regulative development was observed after removal of individual first quartet cells.

In the first polar lobe ofBithynia eggs a special plasm, the vegetal body, is present. It consists of a cupshaped aggregate of small vesicles. Centrifugation of eggs prior to first cleavage may result in displacement of the vegetal body. In about 50% of thecentriguged eggs the vegetal body is found outside the polar lobe, in one of the blastomeres. Removal of the polar lobe from non-centrifuged eggs always leads to severe defects in development. When the lobe is removed from centrifuged eggs, however, about 50% of the eggs develop into normal embryos. It is concluded that the presence of the vegetal body in a blastomere suffices to ensure normal development and, hence, that the polar lobe-specific morphogenetic determinants are contained within the vegetal body.