Atypical APC/C‐dependent degradation of Mcl‐1 provides an apoptotic timer during mitotic arrest

The initiation of apoptosis in response to the disruption of mitosis provides surveillance against chromosome instability. Here, we show that proteolytic destruction of the key regulator Mcl‐1 during an extended mitosis requires the anaphase‐promoting complex or cyclosome (APC/C) and is independent of another ubiquitin E3 ligase, SCF^Fbw7. Using live‐cell imaging, we show that the loss of Mcl‐1 during mitosis is dependent on a D box motif found in other APC/C substrates, while an isoleucine‐arginine (IR) C‐terminal tail regulates the manner in which Mcl‐1 engages with the APC/C, converting Mcl‐1 from a Cdc20‐dependent and checkpoint‐controlled substrate to one that is degraded independently of checkpoint strength. This mechanism ensures a relatively slow but steady rate of Mcl‐1 degradation during mitosis and avoids its catastrophic destruction when the mitotic checkpoint is satisfied, providing an apoptotic timer that can distinguish a prolonged mitotic delay from normal mitosis. Importantly, we also show that inhibition of Cdc20 promotes mitotic cell death more effectively than loss of APC/C activity through differential effects on Mcl‐1 degradation, providing an improved strategy to kill cancer cells.     

Roaming Companion Cats as Potential Causes of Conflict and Controversy: A Representative Questionnaire Study of the Danish Public

Cats have grown in popularity as companion animals, but there are also people who strongly dislike them. Companion cats allowed to roam freely outdoors are seen by some as a nuisance. This paper, drawing on research conducted in Denmark, aims to quantify potentially conflicting attitudes to cats among the public that may feed into cat-related conflicts and controversies. Questionnaire data were collected from a representative sample of the Danish population (n = 2,003), where 21% (n = 415) owned cats and 79% (n = 1,588) did not. In all, 65% of respondents confirmed that they liked cats, 21% reported that they did not, and 14% were undecided. The main reasons for disliking cats concerned “behavior,” not hazards such as the spread of diseases and predation. Of the 21% of the surveyed Danes who reported that they currently had a cat in the household, 72% allowed their cat to roam outdoors. Sixty percent of the respondents did not perceive this as a problem. However, the potential for conflict was demonstrated by the fact that 27% of respondents regarded outdoor roaming as problematic. Of these, about a quarter saw free-roaming cats as a big problem and as a cause of strife between neighbors. Comparatively fewer of those who owned cats saw their animals as a cause of problems. Thus, only 12% of those owning outdoor cats thought that problems were caused when their cats defecated in a neighbor’s garden, which compares with the 17% of the total population who are bothered by other people’s cats defecating in their gardens. Our data show that while the majority of Danes believe cats should be allowed to roam in public spaces, a significant minority strongly dislikes cats and would prefer restrictions on roaming.     

Deregulation of neuronal miRNAs induced by amyloid-β or TAU pathology

Background

Despite diverging levels of amyloid-β (Aβ) and TAU pathology, different mouse models, as well as sporadic AD patients show predictable patterns of episodic memory loss. MicroRNA (miRNA) deregulation is well established in AD brain but it is unclear whether Aβ or TAU pathology drives those alterations and whether miRNA changes contribute to cognitive decline.

Methods

miRNAseq was performed on cognitively intact (4 months) and impaired (10 months) male APPtg (APP^swe/PS1^L166P) and TAUtg (THY-Tau22) mice and their wild-type littermates (APPwt and TAUwt). We analyzed the hippocampi of 12 mice per experimental group (n =?96 in total), and employed a 2-way linear model to extract differentially expressed miRNAs. Results were confirmed by qPCR in a separate cohort of 4 M and 10 M APPtg and APPwt mice (n =?7–9 per group) and in human sporadic AD and non-demented control brain. Fluorescent in situ hybridization identified their cellular expression. Functional annotation of predicted targets was performed using GO enrichment. Behavior of wild-type mice was assessed after intracerebroventricular infusion of miRNA mimics.

Results

Six miRNAs (miR-10a-5p, miR-142a-5p, miR-146a-5p, miR-155-5p, miR-211-5p, miR-455-5p) are commonly upregulated between APPtg and TAUtg mice, and four of these (miR-142a-5p, miR-146a-5p, miR-155-5p and miR-455-5p) are altered in AD patients. All 6 miRNAs are strongly enriched in neurons. Upregulating these miRNAs in wild-type mice is however not causing AD-related cognitive disturbances.

Conclusion

Diverging AD-related neuropathologies induce common disturbances in the expression of neuronal miRNAs. 4 of these miRNAs are also upregulated in AD patients. Therefore these 4 miRNAs (miR-142a-5p, miR-146a-5p, miR-155-5p and miR-455-5p) appear part of a core pathological process in AD patients and APPtg and TAUtg mice. They are however not causing cognitive disturbances in wild-type mice. As some of these miRNA target AD relevant proteins, they may be, in contrast, part of a protective response in AD.

     

Morphological variability of Tragopogon pterodes Pančić ex Petrović achenes: taxonomic evaluation of heterocarpy

This study presents the carpological heterogeneity of Tragopogon pterodes, an endemic plant species from Balkan Peninsula. The goal of this paper was to determine the variability of morphological characters of achenes and assess the degree of their morphological differentiation relative to the position they occupy in the receptacle. Analysis included four different populations of T. pterodes and examination of 18 (8 qualitative and 10 quantitative) characters of achenes in total. Data analysis employed basic statistic, univariate (ANOVA, Tukey HSD) and multivariate (CDA, CA) analyses to check the level of heterocarpy in this species. The results indicate possible morphological differentiation of three groups of achenes (outer, median and inner achenes), depending of their position on the receptacle. The members of all three types were described, illustrated and compared. The variability of investigated carpological characters in this species is providing important information prior to their further taxonomic treatment.     

Behaviour of bovine phosphatidylethanolamine-binding protein with model membranes. Evidence of affinity for negatively charged membranes.

The ability of phosphatidylethanolamine-binding protein (PEBP) to bind membranes was tested by using small and large unilamellar vesicles and monolayers composed of l-alpha-1,2-dimyristoylphosphatidylcholine, l-alpha-1,2-dimyristoylphosphatidylglycerol and l-alpha-1,2-dimyristoylphosphatidylethanolamine. PEBP only bound to model membranes containing l-alpha-1,2-dimyristoylphosphatidylglycerol; the interaction was primarily due to electrostatic forces between the basic protein and the acidic phospholipids. Further experiments indicated that the interaction was not dependent on the length and unsaturation of the phospholipid acyl chains and was not modified by the presence of cholesterol in the membrane. PEBP affinity for negatively charged membranes is puzzling considering the previous identification of the protein as a phosphatidylethanolamine-binding protein, and suggests that the association of PEBP with phospholipid membranes is driven by a mechanism other than its binding to solubilized phosphatidylethanolamine. An explanation was suggested by its three-dimensional structure: a small cavity at the protein surface has been reported to be the binding site of the polar head of phosphatidylethanolamine, while the N-terminal and C-terminal parts of PEBP, exposed at the protein surface, appear to be involved in the interaction with membranes. To test this hypothesis, we synthesized the two PEBP terminal regions and tested them with model membranes in parallel with the whole protein. Both peptides displayed the same behaviour as whole PEBP, indicating that they could participate in the binding of PEBP to membranes. Our results strongly suggest that PEBP directly interacts with negatively charged membrane microdomains in living cells.     

Anthropometric and health-related fitness characteristics in middle-aged obese women

The purpose of this investigation was to determine the possible relationships between anthropometrical and health-related fitness parameters in obese middle-aged women. Twenty one obese (BMI > 27 kg/m2) and 12 control (BMI < 27 kg/m2) middle-aged females (35-45 yrs) participated in this investigation. Three series of anthropometrical measurements on the right side of the body were taken according to the O-scale physique assessment system. The Heath-Carter anthropometric somatotyping method was used and the ratio of waist to hip circumferences was calculated. The body composition was measured using bioelectrical impedance method (Bodystat-500, UK). The following health-related fitness tests used were: dynamic sit-up, hand grip, sit-and-reach, single leg balance and plate tapping. Physical working capacity (PWC) was measured using single ergometer test. Obese women possessed significantly higher (p < 0.05) values for skinfold, girth and breadth measurements. While the differences in somatotype indices were not statistically significant (p > 0.05) between obese and control groups, the transformation of somatotype characteristics to the effect sizes (ESs) revealed that these differences were large (ectomorphy: ES = 1.73; endomorphy: ES = 1.64; mesomorphy: ES = 1.71). Relative aerobic fitness (VO2max/kg, calculated from PWC) and dynamic sit-up were higher (p < 0.05) in control subjects, while obese women presented significantly higher values for hand grip strength. The thicker skinfold thicknesses increased the absolute value of PWC in obese group (r = 0.39-0.57; p < 0.05). In contrast, the thinner skinfold thicknesses in suprailiac and mid-thigh significantly increased the absolute value of PWC in control women. There were only a few significant correlations between girth and breadth measures, and health-related fitness parameters. In addition, somatotype characteristics seldom influenced the results of health-related fitness tests. Stepwise multiple regression analysis demonstrated that health-related fitness test items were more dependent on the anthropometrical parameters in obese than in control women.     

New Technique for Biotyping

An efficient technique for biotyping, using disposable trays instead of glass tubes and agar instead of liquid media, is described.     

Escherichia coli O Antigen Typing by Means of a Mechanized Microtechnique

A mechanized microtechnique for the serological typing of Escherichia coli cultures is described.