Identification of a catalytic active but non-aggregating MDM2 RING domain variant

As a key regulator of the tumour suppressor protein p53, MDM2 is involved in various types of cancer and has thus been an attractive drug target. So far, small molecule design has primarily focussed on the N-terminal p53-binding domain although on-target toxicity effects have been reported. Targeting the catalytic RING domain of MDM2 resembles an alternative approach to drug MDM2 with the idea to prevent MDM2-mediated ubiquitination of p53 while retaining MDM2's ability to bind p53. The design of RING inhibitors has been limited by the extensive aggregation tendency of the RING domain, making it challenging to undertake co-crystallization attempts with potential inhibitors. Here we compare the purification profiles of the MDM2 RING domain from several species and show that the MDM2 RING domain of other species than human is much less prone to aggregate although the overall structure of the RING domain is conserved. Through sequence comparison and mutagenesis analyses, we identify a single point mutation, G443T, which greatly enhances the dimeric fraction of human MDM2 RING domain during purification. Neither does the mutation alter the structure of the RING domain, nor does it affect E2(UbcH5B)-Ub binding and activity. Hence, MDM2-G443T facilitates studies involving binding partners that would be hampered by the low solubility of the wild-type RING domain. Furthermore, it will be valuable for the development of MDM2 RING inhibitors

The influence of spatial pattern on visual short-term memory for contrast

Several psychophysical studies of visual short-term memory (VSTM) have shown high-fidelity storage capacity for many properties of visual stimuli. On judgments of the spatial frequency of gratings, for example, discrimination performance does not decrease significantly, even for memory intervals of up to 30 s. For other properties, such as stimulus orientation and contrast, however, such “perfect storage” behavior is not found, although the reasons for this difference remain unresolved. Here, we report two experiments in which we investigated the nature of the representation of stimulus contrast in VSTM using spatially complex, two-dimensional random-noise stimuli. We addressed whether information about contrast per se is retained during the memory interval by using a test stimulus with the same spatial structure but either the same or the opposite local contrast polarity, with respect to the comparison (i.e., remembered) stimulus. We found that discrimination thresholds got steadily worse with increasing duration of the memory interval. Furthermore, performance was better when the test and comparison stimuli had the same local contrast polarity than when they were contrast-reversed. Finally, when a noise mask was introduced during the memory interval, its disruptive effect was maximal when the spatial configuration of its constituent elements was uncorrelated with those of the comparison and test stimuli. These results suggest that VSTMfor contrast is closely tied to the spatial configuration of stimuli and is not transformed into a more abstract representation

The association between muscular power from childhood to adulthood and adult measures of glucose homeostasis

This study aimed to assess whether the longitudinal association between childhood muscular fitness and adult measures of glucose homeostasis persist despite changes in muscular fitness across the life course. This prospective longitudinal study included 586 participants who had their muscular power (standing long jump distance), cardiorespiratory fitness (CRF), and waist circumference measured as children (aged 9, 12, 15 years) and again 20 years later as adults. In adulthood, these participants also provided a fasting blood sample which was tested for glucose and insulin. Glucose homeostasis measures including insulin resistance (HOMA2-IR) and beta cell function (HOMA2-β) were estimated. Child and adult muscular power levels were separated into thirds, and tracking groups (persistently low, decreasing, persistently moderate, increasing, and persistently high) were created. Sex-stratified multivariable linear regression models were used to examine the association between muscular power tracking groups and adult measures of glucose homeostasis. Compared with males with persistently high muscular power, males with increasing and persistently low muscular power had higher fasting insulin (increasing: β = 1.12 mU/L, P = .04; persistently low: β = 2.12 mU/L, P = .001) and HOMA2-β (increasing: β = 8.50%, P = .03; persistently low: β = 11.27%, P = .01) independent of CRF and males with persistently low muscular power had greater fasting insulin (β = 1.22 mU/L, P = .02) and HOMA2-IR (β = 0.14, P = .02) independent of waist circumference. Non-significant associations were present for females. For males, maintaining persistently high muscular power between childhood and adulthood could lead to a healthier adult glucose homeostasis profile

The association between grip strength measured in childhood, young- and mid-adulthood and prediabetes or type 2 diabetes in mid-adulthood

Background: Although low child and adult grip strength is associated with adverse cardiometabolic health, how grip strength across the life course associates with type 2 diabetes is unknown. This study identified the relative contribution of grip strength measured at specific life stages (childhood, young adulthood, mid-adulthood) with prediabetes or type 2 diabetes in mid-adulthood. Methods: Between 1985 and 2019, 263 participants had their grip strength measured using an isometric dynamometer in childhood (9-15 years), young adulthood (28-36 years) and mid-adulthood (38-49 years). In mid-adulthood, a fasting blood sample was collected and tested for glucose and glycated haemoglobin (HbA1c). Participants were categorized as having prediabetes or type 2 diabetes if fasting glucose levels were ≥ 5.6 mmol or if HbA1c levels were ≥ 5.7% (≥ 39 mmol/mol). A Bayesian relevant life course exposure model examined the association between lifelong grip strength and prediabetes or type 2 diabetes. Results: Grip strength at each time point was equally associated with prediabetes or type 2 diabetes in mid-adulthood (childhood: 37%, young adulthood: 36%, mid-adulthood: 28%). A one standard deviation increase in cumulative grip strength was associated with 34% reduced odds of prediabetes or type 2 diabetes in mid-adulthood (OR 0.66, 95% credible interval 0.40, 0.98). Conclusions: Greater grip strength across the life course could protect against the development of prediabetes and type 2 diabetes. Strategies aimed at increasing muscular strength in childhood and maintaining behaviours to improve strength into adulthood could improve future cardiometabolic health. The Association Between Grip Strength Measured in Childhood, Young- and Mid-adulthood and Prediabetes or Type 2 Diabetes in Mid-adulthood

Treatment of Advanced Emphysema with Emphysematous Lung Sealant (AeriSeal (R))

Background: This report summarizes initial tests of an emphysematous lung synthetic polymer sealant (ELS) designed to reduce lung volume in patients with advanced emphysema. Objectives: The primary study objective was to define a therapeutic strategy to optimize treatment safety and effectiveness. Methods: ELS therapy was administered bronchoscopically to 25 patients with heterogeneous emphysema in an open-label, noncontrolled study at 6 centers in Germany. Treatment was performed initially at 2-4 subsegments. After 12 weeks, patients were eligible for repeat therapy to a total of 6 sites. Safety and efficacy were assessed after 6 months. Responses were evaluated in terms of changes from baseline in lung physiology, functional capacity, and health-related quality of life. Follow-up is available for 21 of 25 patients. Results: Treatment was well tolerated. There were no treatment-related deaths (i.e. within 90 days of treatment), and an acceptable short-and long-term safety profile. Physiological and clinical benefits were observed at 24 weeks. Efficacy responses were better among Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage III patients {[}n = 14; change in residual volume/total lung capacity (Delta RV/TLC) = -7.4 +/- 10.3%; Delta forced expiratory volume in 1 s (Delta FEV(1)) = +15.9 +/- 22.6%; change in forced vital capacity (Delta FVC) = +24.1 +/- 22.7%; change in carbon monoxide lung diffusion capacity (Delta DLCO) = +19.3 +/- 34.8%; change in 6-min walk test (Delta 6MWD) = +28.7 +/- 59.6 m; change in Medical Research Council Dyspnea (Delta MRCD) score = -1.0 +/- 1.04 units; change in St. George's Respiratory Questionnaire (Delta SGRQ) score = -9.9 +/- 15.3 units] than for GOLD stage IV patients (n = 7; Delta RV/TLC = -0.5 +/- 6.4%; Delta FEV 1 = +2.3 +/- 12.3%; Delta FVC = +2.6 +/- 21.1%; Delta DLCO = -2.8 +/- 17.2%; Delta 6MWD = +28.3 +/- 58.4 m; Delta MRCD = 0.3 +/- 0.81 units; Delta SGRQ = -6.7 +/- 7.0 units). Conclusions: ELS therapy shows promise for treating patients with advanced heterogeneous emphysema. Additional studies to assess responses in a larger cohort with a longer follow-up are warranted. Copyright (C) 2011 S. Karger AG, Base

Tracking of secretory phospholipase A2 enzyme activity levels from childhood to adulthood: a 21-year cohort.

OBJECTIVE: Secretory phospholipase A2 (sPLA2) enzyme activity is a potential inflammatory biomarker for cardiovascular disease. We examined the tracking, or persistence, of sPLA2 enzyme activity levels from childhood to adulthood, and identify potentially modifiable factors affecting tracking. METHOD: Prospective cohort of 1735 children (45% females) who had serum sPLA2 enzyme activity levels and other cardiovascular disease risk factors measured in 1980 that were followed-up in 2001. RESULTS: sPLA2 activity tracked from childhood to adulthood for males (r=0.39) and females (r=0.45). Those who decreased body mass index relative to their peers were more likely to resolve elevated childhood sPLA2 levels than have persistent elevated sPLA2 levels in childhood and adulthood. Those who consumed less fruit, and gained more body mass index relative to their peers, began smoking or were a persistent smoker between childhood and adulthood were more likely to develop incident elevated sPLA2 levels than those with persistent not elevated sPLA2 levels. CONCLUSIONS: Childhood sPLA2 enzyme activity levels associate with adult sPLA2 levels 21 years later. Healthful changes in modifiable risk factors that occur between childhood and adulthood might prevent children from developing elevated sPLA2 levels in adulthood

C-reactive protein and albumin kinetics before community-acquired bloodstream infections- A Danish population-based cohort study

aEarly changes in biomarker levels probably occur before bloodstream infection (BSI) is diagnosed. However, this issue has not been fully addressed. We aimed at evaluating the kinetics of C-reactive protein (CRP) and plasma albumin (PA) in the 30 days before community-acquired (CA) BSI diagnosis. From a population-based BSI database we identified 658 patients with at least one measurement of CRP or PA from day-30 (D-30) through day-1 (D-1) before the day of CA-BSI (D0) and a measurement of the same biomarker at D0 or D1. Amongst these, 502 had both CRP and PA measurements which fitted these criteria. CRP and PA concentrations began to change inversely some days before CA-BSI diagnosis, CRP increasing by day-3.1 and PA decreasing by day-1.3. From D-30 to D-4, CRP kinetics (expressed as slopes-rate of concentration change per day) was-1.5 mg/l/day. From D-3 to D1, the CRP slope increased to 36.3 mg/l/day. For albumin, the slope between D-30 to D-2 was 0.1 g/l/day and changed to-1.8 g/l/day between D-1 and D1. We showed that biomarker levels begin to change some days before the CA-BSI diagnosis, CRP 3.1 days and PA 1.3 days before.publishersversionepub_ahead_of_prin

Analysis of periosteal lesions from commingled human remains at the Xagħra Circle hypogeum reveals the first case of probable scurvy from Neolithic Malta

Abstract Objectives Palaeopathological analysis is key for characterising population health at the individual level and across large assemblages but is rarely exploited to unite the remains of disarticulated individuals. This study explores the potential for individual identification through differential diagnosis of periosteal lesions in a commingled deposit, both to ascertain the number of individuals represented and provide a differential diagnosis. Materials and Methods The late Neolithic Xag?ra Circle hypogeum on Gozo contains the remains of more than 800 individuals, most of which were transformed to a collective disarticulated assemblage. Across the excavated population, pathological observations are strikingly low. In one specific 1???1-m area in a single stratigraphic context, fragmented and disarticulated cranial and post-cranial non-adult bones were identified that displayed periosteal new bone formation. To aid differential diagnosis, macroscopic analysis, taphonomic analysis and micro-computed tomography (?CT) imaging were integrated. Results This approach, when combined with osteobiographical analyses, reveals that the elements most likely derive from one individual, a young child, who presents a probable case of scurvy. The potential for micronutrient co-morbidities are explored, but without further microscopic study it cannot be determined if this individual also experienced iron-deficiency anaemia and/or rickets. Discussion In the context of the Mediterranean and Europe in later prehistory, reported cases of scurvy are currently low and often reveal periods of environmental instability and resource insufficiency. Our finding of non-adult scurvy in late 3rd millennium BC Malta contributes to a developing picture of an increasingly unstable palaeoenvironment and declining population health at this time, although it may also indicate an individual case of poor childhood health within this broader context.1 Introduction 1.1 Archaeological context of the Xagħra Circle hypogeum 2 Materials and methods 2.1 Remains presenting periosteal lesions 2.2 Macroscopic and micro-CT analysis 3 Macroscopic and radiological observations of pathology 3.1 Frontal bone (FB0039, FB0040) and zygoma (FB0041) 3.2 Mandible (FB0042) 3.3 Left rib (FB0043) and right rib (FB0044) 3.4 Ulna (FB0045) 4 Differential diagnosis 4.1 Frontal bone (FB0039, FB0040) and zygoma (FB0041) 4.2 Mandible (FB0042) 4.3 Left rib (FB0043) and right rib (FB0044) 4.4 Ulna (FB0045) 5 Discussion 5.1 Scurvy: Causes, consequences and comorbidities 5.2 Scurvy in prehistory 5.3 The Maltese context 6 Conclusio

Childhood Infections, Socioeconomic Status, and Adult Cardiometabolic Risk

BACKGROUND AND OBJECTIVES: Socioeconomic disadvantage throughout the life course is associated with increased risk of cardiometabolic diseases, but traditional risk factors do not fully account for the social gradient. We investigated the interactions between low socioeconomic status (SES) and infection in childhood and adverse cardiometabolic parameters in adulthood. METHODS: Participants from the Cardiovascular Risk in Young Finns Study, a cohort well phenotyped for childhood and adulthood cardiometabolic risk factors and socioeconomic parameters, were linked to lifetime hospitalization data from birth onward available from the Finnish National Hospital Registry. In those with complete data, we investigated relationships between infection-related hospitalization in childhood, SES, and childhood and adult cardiometabolic parameters. RESULTS: The study cohort consisted of 1015 participants (age range 3–18 years at baseline and 30–45 years at follow-up). In adults who were raised in below-median income families, childhood infection-related hospitalizations (at age 0–5 years) were significantly associated with higher adult BMI (β ± SE comparing those with 0 vs ≥1 hospitalizations 2.4 ± 0.8 kg/m2, P = .008), waist circumference (7.4 ± 2.3 cm, P = .004), and reduced brachial flow–mediated dilatation (−2.7 ± 0.9%, P = .002). No equivalent associations were observed in participants from higher-SES families. CONCLUSIONS: Infection was associated with worse cardiovascular risk factor profiles only in those from lower-SES families. Childhood infection may contribute to social gradients observed in adult cardiometabolic disease risk factors. These findings suggest reducing childhood infections, especially in socioeconomic disadvantaged children, may reduce the cardiometabolic disease burden in adults