MDA-MB-231 breast cancer cell viability,motility and matrix adhesion are regulated by a complex interplay of heparan sulfate,chondroitin−/dermatan sulfate and hyaluronan biosynthesis

Proteoglycans and glycosaminoglycans modulate numerous cellular processes relevant to tumour progression, including cell proliferation, cell-matrix interactions, cell motility and invasive growth. Among the glycosaminoglycans with a well-documented role in tumour progression are heparan sulphate, chondroitin/dermatan sulphate and hyaluronic acid/hyaluronan. While the mode of biosynthesis differs for sulphated glycosaminoglycans, which are synthesised in the ER and Golgi compartments, and hyaluronan, which is synthesized at the plasma membrane, these polysaccharides partially compete for common substrates. In this study, we employed a siRNA knockdown approach for heparan sulphate (EXT1) and heparan/chondroitin/dermatan sulphate-biosynthetic enzymes (β4GalT7) in the aggressive human breast cancer cell line MDA-MB-231 to study the impact on cell behaviour and hyaluronan biosynthesis. Knockdown of β4GalT7 expression resulted in a decrease in cell viability, motility and adhesion to fibronectin, while these parameters were unchanged in EXT1-silenced cells. Importantly, these changes were associated with a decreased expression of syndecan-1, decreased signalling response to HGF and an increase in the synthesis of hyaluronan, due to an upregulation of the hyaluronan synthases HAS2 and HAS3. Interestingly, EXT1-depleted cells showed a downregulation of the UDP-sugar transporter SLC35D1, whereas SLC35D2 was downregulated in β4GalT7-depleted cells, indicating an intricate regulatory network that connects all glycosaminoglycans synthesis. The results of our in vitro study suggest that a modulation of breast cancer cell behaviour via interference with heparan sulphate biosynthesis may result in a compensatory upregulation of hyaluronan biosynthesis. These findings have important implications for the development of glycosaminoglycan-targeted therapeutic approaches for malignant diseases.     

Reconstruction of radionuclide intakes for the residents of East Urals Radioactive Trace (1957–2011)

The East Urals Radioactive Trace (EURT) was formed after a chemical explosion in the radioactive waste-storage facility of the Mayak Production Association in 1957 (Southern Urals, Russia) and resulted in an activity dispersion of 7.4?×?10¹⁶ Bq into the atmosphere. Internal exposure due to ingestion of radionuclides with local foodstuffs was the main factor of public exposure at the EURT. The EURT cohort, combining residents of most contaminated settlements, was formed for epidemiological study at the Urals Research Center for Radiation Medicine, Russia (URCRM). For the purpose of improvement of radionuclide intake estimates for cohort members, the following data sets collected in URCRM were used: (1) Total β-activity and radiochemical measurements of ⁹⁰Sr in local foodstuffs over all of the period of interest (1958–2011; n?=?2200), which were used for relative ⁹⁰Sr intake estimations. (2) ⁹⁰Sr measurements in human bones and whole body (n?=?338); these data were used for average ⁹⁰Sr intake derivations using an age- and gender-dependent Sr-biokinetic model. Non-strontium radionuclide intakes were evaluated on the basis of ⁹⁰Sr intake data and the radionuclide composition of contaminated foodstuffs. Validation of radionuclide intakes during the first years after the accident was first carried out using measurements of the feces β-activity of EURT residents (n?=?148). The comparison of experimental and reconstructed values of feces β-activity shows good agreement. ⁹⁰Sr intakes for residents of settlements evacuated 7–14 days after the accident were also obtained from ⁹⁰Sr measurements in human bone and whole body. The results of radionuclide intake reconstruction will be used to estimate the internal doses for the members of the EURT cohort.     

Heating-induced transition of Potyvirus Potato Virus A coat protein into β-structure

In our previous communication, we have reported that virions of plant Potyvirus Potato Virus A (PVA) have a peculiar structure characterized by high content of disordered regions in intravirus coat protein (CP). In this report, we describe unusual properties of the PVA CP. With the help of a number of physicochemical methods, we have observed that the PVA CP just released from the virions by heating at 60–70 °C undergoes association into oligomers and transition to β- (and even cross-β-) conformation. Transition to β-structure on heating has been recently reported for a number of viral and non-viral proteins. The PVA CP isolated by LiCl method was also transformed into cross-β-structure on heating to 60 °C. Using the algorithms for protein aggregation prediction, we found that the aggregation-prone segments should be located in the central region of a PVA CP molecule. Possibly this transition mimics some functions of PVA CP in the virus life cycle in infected plants.     

Mortality among patients with frequent emergency department use for alcohol-related reasons in Ontario: a population-based cohort study

BACKGROUND:Little is known about the risk of death among people who visit emergency departments frequently for alcohol-related reasons, including whether mortality risk increases with increasing frequency of visits. Our primary objective was to describe the sociodemographic and clinical characteristics of this high-risk population and examine their 1-year overall mortality, premature mortality and cause of death as a function of emergency department visit frequency in Ontario, Canada.METHODS:We conducted a population-based retrospective cohort study using linked health administrative data (Jan. 1, 2010, to Dec. 31, 2016) in Ontario for people aged 16–105 years who made at least 2 emergency department visits for mental or behavioural disorders due to alcohol within 1 year. We subdivided the cohort based on visit frequency (2, 3 or 4, or ≥ 5). The primary outcome was 1-year mortality, adjusted for age, sex, income, rural residence and presence of comorbidities. We examined premature mortality using years of potential life lost (YPLL).RESULTS:Of the 25 813 people included in the cohort, 17 020 (65.9%) had 2 emergency department visits within 1 year, 5704 (22.1%) had 3 or 4 visits, and 3089 (12.0%) had 5 or more visits. Males, people aged 45–64 years, and those living in urban centres and lower-income neighbourhoods were more likely to have 3 or 4 visits, or 5 or more visits. The all-cause 1-year mortality rate was 5.4% overall, ranging from 4.7% among patients with 2 visits to 8.8% among those with 5 or more visits. Death due to external causes (e.g., suicide, accidents) was most common. The adjusted mortality rate was 38% higher for patients with 5 or more visits than for those with 2 visits (adjusted hazard ratio 1.38, 95% confidence interval 1.19–1.59). Among 25 298 people aged 16–74 years, this represented 30 607 YPLL.INTERPRETATION:We observed a high mortality rate among relatively young, mostly urban, lower-income people with frequent emergency department visits for alcohol-related reasons. These visits are opportunities for intervention in a high-risk population to reduce a substantial mortality burden.

Alcohol is a leading driver of morbidity and mortality worldwide.¹ An estimated 3 million deaths in 2016 — 5% of all global deaths — were attributable to alcohol consumption.² The 2016 Global Burden of Disease Study showed that alcohol was the single greatest risk factor for ill health worldwide among people aged 15–49 years.³ In Canada, hospital admissions for alcohol-attributable conditions out-number those for myocardial infarction.⁴ Alcohol-related harms cost Canadians about $14.6 billion annually, with $3.3 billion in health care costs.⁵In addition to the societal impact of mental and behavioural disorders due to alcohol (henceforth referred to as alcohol-related) — mainly acute intoxication and withdrawal — these disorders are common reasons for emergency department visits.^6,7 Data from the United States and Canada, furthermore, suggest that alcohol-related emergency department visits have increased in recent years.^8,9 For example, a study in Ontario showed that, between 2003 and 2016, the age-standardized rates of alcohol-attributable emergency department visits increased by 86.5% in women and 53.2% in men.⁸ People who visit emergency departments frequently for alcohol-related reasons have high levels of comorbidity and social disadvantage,^10,11 and represent a readily identifiable patient population for whom interventions to address unmet social and health care needs could be developed.^12–14 A systematic review suggested that screening and brief intervention for alcohol-related problems in the emergency department is a promising approach for reducing problematic alcohol consumption.¹³Despite this, little is known about the risk of death, a key outcome for health system performance, among people who use emergency departments frequently for alcohol-related reasons, including whether mortality risk increases with increasing frequency of visits. To address this gap, our primary objective was to describe the sociodemographic and clinical characteristics of this high-risk population and examine their 1-year overall mortality, premature mortality and cause of death as a function of emergency department visit frequency in Ontario, the most populous Canadian province.¹⁵     

Evidence that gestation duration and lactation duration are coupled traits in primates

Gestation duration and lactation duration are usually treated as independently evolving traits in primates, but the metabolic theory of ecology (MTE) suggests both durations should be determined by metabolic rate. We used phylogenetic generalized least-squares linear regression to test these different perspectives. We found that the allometries of the durations are divergent from each other and different from the scaling exponent predicted by the MTE (0.25). Gestation duration increases much more slowly (0.06 < m < 0.12), and lactation duration much more quickly (0.36 < m < 0.52) with body mass than the MTE predicts. By contrast, we found that the combined duration of gestation and lactation is consistent with the MTE''s predictions (0.22 < m < 0.35). These results suggest that gestation duration and lactation duration might best be viewed as distinct but coupled adaptations. When transferring energy to their offspring, primate mothers must meet metabolically dictated physiological requirements while optimizing the timing of the switch from gestation to lactation in relation to some as-yet-unidentified body-size-related factor.     

Eucaryotic cell components that bind to chlamydial elementary bodies: the histones

HeLa-cell-membrane fractions isolated by sonication as used previously to identify chlamydial adhesins were examined by a blotting technique for binding chlamydial elementary bodies (EB). One HeLa cell protein with apparent molecular mass of 32 kDa was found to bind native EB. A monoclonal antibody (mAb) raised against this chlamydial binding host-cell protein reacted with eucaryotic histones. Histone fractions were capable of binding EB in an ELISA assay and histone H1 was identified as the chlamydial-binding host cell protein in the Hela cell membrane fraction. Probing with specific mAbs against histone H3 and DNA confirmed that chromatin components were present in the host-cell membrane extract. These data suggest that the HeLa-cell-binding chlamydial proteins were previously identified by their reaction with chromatin and not with membrane components.     

Increase in accumulation of strontium-90 in the maternal skeleton during pregnancy and lactation: analysis of the Techa River data

The unique contamination of the Techa River (Southern Urals, Russia) in the 1950s by long-lived ⁹⁰Sr allows investigation of the accumulation of bone-seeking elements in humans. This study is based on information compiled at the Urals Research Center for Radiation Medicine (Chelyabinsk, Russia) over a long period of time. It includes the results of in vivo measurements of ⁹⁰Sr-body burden with a whole body counter (WBC), data on personal medical examinations and residence and family histories. Data on 185 women from two Techa riverside villages Muslyumovo and Brodokalmak were selected. The settlements differ in terms of ⁹⁰Sr diet intake (higher in Muslyumovo than in Brodokalmak) and ethnicity (residents were mainly Slavs in Brodokalmak and Turkic in Muslyumovo). Results of a total of 555 WBC measurements performed in 1974–1997 were available for the women studied; maximum measured values reached 40 kBq/body. The women from each settlement were subdivided into three groups according to their childbearing history: pregnancy and lactation occurred (1) during the period of maximal ⁹⁰Sr intake (1950–1951); (2) after the period of maximal intake and (3) before this period or women who were childless. An increase was found in accumulation of ⁹⁰Sr in maternal skeleton during pregnancy and lactation (group 1) by a factor of 1.5–2 in comparison with non-pregnant, non-lactating women. This result was found in both Muslyumovo and Brodokalmak samples. An increase in accumulation of toxic elements in pregnant/lactating women is associated with increased radiation/toxic doses and risk for the women’s health.     

Preparation, resonance assignment, and preliminary dynamics characterization of residue specific 13C/15N-labeled elongated DNA for the study of sequence-directed dynamics by NMR

DNA is a highly flexible molecule that undergoes functionally important structural transitions in response to external cellular stimuli. Atomic level spin relaxation NMR studies of DNA dynamics have been limited to short duplexes in which sensitivity to biologically relevant fluctuations occurring at nanosecond timescales is often inadequate. Here, we introduce a method for preparing residue-specific ¹³C/¹⁵N-labeled elongated DNA along with a strategy for establishing resonance assignments and apply the approach to probe fast inter-helical bending motions induced by an adenine tract. Preliminary results suggest the presence of elevated A-tract independent end-fraying internal motions occurring at nanosecond timescales, which evade detection in short DNA constructs and that penetrate deep (7 bp) within the DNA helix and gradually fade away towards the helix interior.