The dynamics of cardiolipin synthesis post-mitochondrial fusion

AbstractAlteration in mitochondrial fusion may regulate mitochondrial metabolism. Since the phospholipid cardiolipin (CL) is required for function of the mitochondrial respiratory chain, we examined the dynamics of CL synthesis in growing Hela cells immediately after and 12h post-fusion. Cells were transiently transfected with Mfn-2, to promote fusion, or Mfn-2 expressing an inactive GTPase for 24h and de novo CL biosynthesis was examined immediately after or 12h post-fusion. Western blot analysis confirmed elevated Mfn-2 expression and electron microscopic analysis revealed that Hela cell mitochondrial structure was normal immediately after and 12h post-fusion. Cells expressing Mfn-2 exhibited reduced CL de novo biosynthesis from [1,3-3H]glycerol immediately after fusion and this was due to a decrease in phosphatidylglycerol phosphate synthase (PGPS) activity and its mRNA expression. In contrast, 12h post-mitochondrial fusion cells expressing Mfn-2 exhibited increased CL de novo biosynthesis from [1,3-3H]glycerol and this was due to an increase in PGPS activity and its mRNA expression. Cells expressing Mfn-2 with an inactive GTPase activity did not exhibit alterations in CL de novo biosynthesis immediately after or 12h post-fusion. The Mfn-2 mediated alterations in CL de novo biosynthesis were not accompanied by alterations in CL or monolysoCL mass. [1-14C]Oleate incorporation into CL was elevated at 12h post-fusion indicating increased CL resynthesis. The reason for the increased CL resynthesis was an increased mRNA expression of tafazzin, a mitochondrial CL resynthesis enzyme. Ceramide-induced expression of PGPS in Hela cells or in CHO cells did not alter expression of Mfn-2 indicating that Mfn-2 expression is independent of altered CL synthesis mediated by elevated PGPS. In addition, Mfn-2 expression was not altered in Hela cells expressing phospholipid scramblase-3 or a disrupted scramblase indicating that proper CL localization within mitochondria is not essential for Mfn-2 expression. The results suggest that immediately post-mitochondrial fusion CL de novo biosynthesis is “slowed down” and then 12h post-fusion it is “upregulated”. The implications of this are discussed

The effects of carbogen, carbon dioxide, and oxygen on noise-induced hearing loss

An investigation into the effect of Carbogen (95% O2/5% CO2), 5% CO2/air, and 100% oxygen on cochlear threshold shifts caused by noise was undertaken. Five groups of eight pigmented guinea pigs were exposed to 105 dB broad band noise for 6 h per day for five consecutive days with each group receiving the various gaseous mixtures either during noise exposure or for 1 h immediately after noise exposure. A control group received the same noise exposure but respired air. Auditory threshold shifts, as measured by the auditory evoked brainstem response, were measured at 2,4,8,12,16, 20 and 24 kHz. Recordings were taken pre-exposure and at Day 1, 3, 5, and Weeks 2 and 3 after noise exposure. Carbogen, given during noise exposure, resulted in a trend toward less post noise exposure threshold shift (as compared to controls) which reached statistical significance by Week 3 at all frequencies except 2 and 20 kHz. Subjects given Carbogen after exposure also showed a general trend toward decreased noise induced threshold shifts, as compared to controls, but this was not statistically significant. The mixture of 5% CO2 /air given during noise exposure yielded no difference in threshold shifts as compared to controls. When 100% oxygen was administered during noise exposure, a marked decrease in noise induced threshold shifts could be seen as compared to controls, with differences reaching statistical significance by day 5 at most frequencies. These results indicate that oxygen (i.e. cochlear-oxygenation) is a more important factor than CO2 (i.e., as a vasodilator) in protection of the cochlea from noise induced damage.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/29055/1/0000088.pd

Production of extended-spectrum β -lactamases and the potential indirect pathogenic role of Prevotella isolates from the cystic fibrosis respiratory microbiota

Extended-spectrum β-lactamase (ESBL) production and the prevalence of the β-lactamase-encoding gene blaTEM were determined in Prevotella isolates (n = 50) cultured from the respiratory tract of adults and young people with cystic fibrosis (CF). Time–kill studies were used to investigate the concept of passive antibiotic resistance and to ascertain whether a β-lactamase-positive Prevotella isolate can protect a recognised CF pathogen from the action of ceftazidime in vitro. The results indicated that approximately three-quarters (38/50; 76%) of Prevotella isolates produced ESBLs. Isolates positive for ESBL production had higher minimum inhibitory concentrations (MICs) of β-lactam antibiotics compared with isolates negative for production of ESBLs (P < 0.001). The blaTEM gene was detected more frequently in CF Prevotella isolates from paediatric patients compared with isolates from adults (P = 0.002), with sequence analysis demonstrating that 21/22 (95%) partial blaTEM genes detected were identical to blaTEM-116. Furthermore, a β-lactamase-positive Prevotella isolate protected Pseudomonas aeruginosa from the antimicrobial effects of ceftazidime (P = 0.03). Prevotella isolated from the CF respiratory microbiota produce ESBLs and may influence the pathogenesis of chronic lung infection via indirect methods, including shielding recognised pathogens from the action of ceftazidime

Studying feasibility and effects of a two-stage nursing staff training in residential geriatric care using a 30 month mixed-methods design [ISRCTN24344776]

<p>Abstract</p> <p>Background</p> <p>Transfer techniques and lifting weights often cause back pain and disorders for nurses in geriatric care. The Kinaesthetics care conception claims to be an alternative, yielding benefits for nurses as well as for clients.</p> <p>Starting a multi-step research program on the effects of Kinaesthetics, we assess the feasibility of a two-stage nursing staff training and a pre-post research design. Using quantitative and qualitative success criteria, we address mobilisation from the bed to a chair and backwards, walking with aid and positioning in bed on the staff level as well as on the resident level. In addition, effect estimates should help to decide on and to prepare a controlled trial.</p> <p>Methods/Design</p> <p>Standard basic and advanced Kinaesthetics courses (each comprising four subsequent days and an additional counselling day during the following four months) are offered to n = 36 out of 60 nurses in a residential geriatric care home, who are in charge of 76 residents. N = 22 residents needing movement support are participating to this study.</p> <p>On the staff level, measurements include focus group discussions, questionnaires, physical strain self-assessment (Borg scale), video recordings and external observation of patient assistance skills using a specialised instrument (SOPMAS). Questionnaires used on the resident level include safety, comfort, pain, and level of own participation during mobilisation. A functional mobility profile is assessed using a specialised test procedure (MOTPA).</p> <p>Measurements will take place at baseline (T0), after basic training (T1), and after the advanced course (T2). Follow-up focus groups will be offered at T1 and 10 months later (T3).</p> <p>Discussion</p> <p>Ten criteria for feasibility success are established before the trial, assigned to resources (missing data), processes (drop-out of nurses and residents) and science (minimum effects) criteria. This will help to make rational decision on entering the next stage of the research program.</p> <p>Trial Registration</p> <p>Current Controlled Trials <a href="http://www.controlled-trials.com/ISRCTN24344776">ISRCTN24344776</a>.</p

Microtearding mode study in NSTX using machine learning enhanced reduced model

This article presents a survey of NSTX cases to study the microtearing mode (MTM) stabilities using the newly developed global reduced model for Slab-Like Microtearing modes (SLiM). A trained neutral network version of SLiM enables rapid assessment (0.05s/mode) of MTM with $98\%$ accuracy providing an opportunity for systemic equilibrium reconstructions based on the matching of experimentally observed frequency bands and SLiM prediction across a wide range of parameters. Such a method finds some success in the NSTX discharges, the frequency observed in the experiment matches with what SLiM predicted. Based on the experience with SLiM analysis, a workflow to estimate the potential MTM frequency for a quick assessment based on experimental observation has been established

Spitzer-selected z > 1.3 protocluster candidates in the LSST deep-drilling fields

We have identified 189 candidate z > 1.3 protoclusters and clusters in the LSST Deep Drilling Fields. This sample will enable the measurement of the metal enrichment and star formation history of clusters during their early assembly period through the direct measurement of the rate of supernovae identified through the LSST. The protocluster sample was selected from galaxy overdensities in a Spitzer/IRAC colour-selected sample using criteria that were optimized for protocluster purity using a realistic light-cone. Our tests reveal that 60–80 per cent of the identified candidates are likely to be genuine protoclusters or clusters, which is corroborated by a ∼4σ stacked X-ray signal from these structures. We provide photometric redshift estimates for 47 candidates which exhibit strong peaks in the photo-z distribution of their candidate members. However, the lack of a photo-z peak does not mean a candidate is not genuine, since we find a stacked X-ray signal of similar significance from both the candidates that exhibit photo-z peaks and those that do not. Tests on the light-cone reveal that our pursuit of a pure sample of protoclusters results in that sample being highly incomplete (∼ 4 per cent) and heavily biased towards larger, richer, more massive, and more centrally concentrated protoclusters than the total protocluster population. Most (∼ 75 per cent) of the selected protoclusters are likely to have a maximum collapsed halo mass of between 1013 and 1014 M, with only ∼ 25 per cent likely to be collapsed clusters above 1014 M. However, the aforementioned bias ensures our sample is ∼ 50 per cent complete for structures that have already collapsed into clusters more massive than 1014 M

Prospective Study Examining Clinical Outcomes Associated with a Negative Pressure Wound Therapy System and Barker’s Vacuum Packing Technique

Background The open abdomen has become a common procedure in the management of complex abdominal problems and has improved patient survival. The method of temporary abdominal closure (TAC) may play a role in patient outcome. Methods A prospective, observational, open-label study was performed to evaluate two TAC techniques in surgical and trauma patients requiring open abdomen management: Barker’s vacuum-packing technique (BVPT) and the ABTheraTM open abdomen negative pressure therapy system (NPWT). Study endpoints were days to and rate of 30-day primary fascial closure (PFC) and 30-day all-cause mortality. Results Altogether, 280 patients were enrolled from 20 study sites. Among them, 168 patients underwent at least 48 hours of consistent TAC therapy (111 NPWT, 57 BVPT). The two study groups were well matched demographically. Median days to PFC were 9 days for NPWT versus 12 days for BVPT (p = 0.12). The 30-day PFC rate was 69 % for NPWT and 51 % for BVPT (p = 0.03). The 30-day all-cause mortality was 14 % for NPWT and 30 % for BVPT (p = 0.01). Multivariate logistic regression analysis identified that patients treated with NPWT were significantly more likely to survive than the BVPT patients [odds ratio 3.17 (95 % confidence interval 1.22–8.26); p = 0.02] after controlling for age, severity of illness, and cumulative fluid administration. Conclusions Active NPWT is associated with significantly higher 30-day PFC rates and lower 30-day all-cause mortality among patients who require an open abdomen for at least 48 h during treatment for critical illness