The relationship between physical activity, appetite and energy intake in older adults: A systematic review.

Ageing often causes a reduction in appetite and energy intake in older adults which can result in malnutrition. Current guidelines for older adults suggest increasing physical activity to enhance appetite. However, it is unclear if there is evidence to support this advice. This aim of this review is to assess if appetite and energy intake is changed in older adults that undertake acute or regular physical activity (measured from cross sectional and intervention studies). Databases SPORTDiscus, CINAHL, MEDLINE were searched for studies between 1970 and 2017 using search terms related to ageing, physical activity (including exercise), energy and appetite. Studies included contained adults over 60 years, including acute, cross-sectional and intervention (longitudinal) studies. Of 34 full-text articles assessed, 8 were included. The Cochrane Collaboration's tool was used for assessing risk of bias. No acute studies were found. Of the cross-sectional studies, one study suggested that individuals who undertake habitual physical activity had an increased energy intake and none of the studies found differences in appetite ratings. Energy intakes increased in the intervention studies, though not always sufficiently to maintain energy balance. One study showed that ability to correctly compensate for previous energy intake was better in those that undertake habitual physical activity. The limited number of studies, wide range of data collection methodologies, time-scales and interventions mean that definitive outcomes are difficult to identify. At this stage advice to increase acute or habitual physical activity as a mean to increase appetite is not supported by sufficient evidence.

A protocol for a randomised double-blind placebo-controlled feasibility study to determine whether the daily consumption of flavonoid-rich pure cocoa has the potential to reduce fatigue in people with relapsing and remitting multiple sclerosis (RRMS).

BACKGROUND: Dietary interventions including consumption of flavonoids, plant compounds found in certain foods, may have the ability to improve fatigue. However, to date, no well-designed intervention studies assessing the role of flavonoid consumption for fatigue management in people with MS (pwMS) have been performed. The hypothesis is that the consumption of a flavonoid-rich pure cocoa beverage will reduce fatigue in pwMS. The aim of this study is to determine the feasibility and potential outcome of running a trial to evaluate this hypothesis. METHODS: Using a randomised (1:1) double-blind placebo-controlled feasibility study, 40 men and women (20 in each trial arm) with a recent diagnosis (< 10 years) of relapsing and remitting MS (RRMS) and who are over 18 years of age will be recruited from neurology clinics and throughout the Thames Valley community. During a 6-week nutrition intervention period, participants will consume the cocoa beverage, high flavonoid or low flavonoid content, at breakfast daily. At baseline, demographic factors and disease-related factors will be assessed. Fatigue, activity and quality of life, in addition to other measures, will be taken at three visits (baseline, week 3 and week 6) in a university setting by a researcher blinded to group membership. Feasibility and fidelity will be assessed through recruitment and retention, adherence and a quantitative process evaluation at the end of the trial.We will describe demographic factors (age, gender, level of education) as well as disease-related factors (disease burden scores, length of time diagnosed with MS) and cognitive assessment, depression and quality of life and general physical activity in order to characterise participants and determine possible mediators to identify the processes by which the intervention may bring about change. Feasibility (recruitment, safety, feasibility of implementation of the intervention and evaluation, protocol adherence and data completion) and potential for benefit (estimates of effect size and variability) will be determined to inform future planned studies. Results will be presented using point estimates, 95% confidence intervals and p values. Primary statistical analysis will be on an intention-to-treat basis and will use the complete case data set. DISCUSSION: We propose that a flavonoid-enriched cocoa beverage for the management of fatigue will be well received by participants. Further, if it is implemented early in the disease course of people diagnosed with RRMS, it will improve mobility and functioning by modifying fatigue. TRIAL REGISTRATION: Registered with ISRCTN Registry. Trial registration No: ISRCTN69897291; Date April 2016.

Coconut oil has less satiating properties than medium chain triglyceride oil.

It is well established that the consumption of medium-chain triglycerides (MCT) can increase satiety and reduce food intake. Many media articles promote the use of coconut oil for weight loss advocating similar health benefits to that of MCT. The aim of this study was to examine the effect of MCT oil compared to coconut oil and control oil on food intake and satiety. Following an overnight fast, participants consumed a test breakfast smoothie containing 205kcal of either (i) MCT oil (ii) coconut oil or (iii) vegetable oil (control) on three separate test days. Participants recorded appetite ratings on visual analogue scales and were presented with an ad libitum lunch meal of preselected sandwiches 180min after consumption of the breakfast. The results showed a significant difference in energy and macronutrient intakes at the ad libitum meal between the three oils with the MCT oil reducing food intake compared to the coconut and control oil. Differences in food intake throughout the day were found for energy and fat, with the control having increased food intake compared to the MCT and coconut. The MCT also increased fullness over the three hours after breakfast compared to the control and coconut oils. The coconut oil was also reported as being less palatable than the MCT oil. The results of this study confirm the differences that exist between MCT and coconut oil such that coconut oil cannot be promoted as having similar effects to MCT oil on food intake and satiety.

They say coconut oil can aid weight loss, but can it really?

There has in recent years, been much media speculation and consumer interest in the beneficial satiating properties of consuming coconut oil and its potential to aid weight loss. However, the media has primarily cited studies using medium-chain triglycerides (MCT) oil. The current perspective looks at the research that is available on coconut oil. It examines if and how MCT-related research can be applied to coconut oil and if there is potential for coconut oil to aid weight loss. The current report indicates a lack of consistent evidence on the topic of coconut oil, satiety and weight loss. Given both the publicity and the increased consumption of coconut oil further research, particularly long-term clinical trials, in this area are warranted.

Applying an extended theory of planned behaviour to predict breakfast consumption in adolescents.

BACKGROUND/OBJECTIVES: Breakfast skipping increases during adolescence and is associated with lower levels of physical activity and weight gain. Theory-based interventions promoting breakfast consumption in adolescents report mixed findings, potentially because of limited research identifying which determinants to target. This study aimed to: (i) utilise the Theory of Planned Behaviour (TPB) to identify the relative contribution of attitudes (affective, cognitive and behavioural) to predict intention to eat breakfast and breakfast consumption in adolescents and (ii) determine whether demographic factors moderate the relationship between TPB variables, intention and behaviour. SUBJECTS/METHODS: Questionnaires were completed by 434 students (mean 14±0.9 years) measuring breakfast consumption (0-2, 3-6 or 7 days), physical activity levels and TPB measures. Data were analysed by breakfast frequency and demographics using hierarchical and multinomial regression analyses. RESULTS: Breakfast was consumed everyday by 57% of students, with boys more likely to eat a regular breakfast, report higher activity levels and report more positive attitudes towards breakfast than girls (P<0.001). The TPB predicted 58% of the variation in intentions. Overall, the model was predictive of breakfast behaviours (P<0.001), but the relative contribution of TPB constructs varied depending on breakfast frequency. Interactions between gender and intentions were significant when comparing 0-2- and 3-6-day breakfast eaters only highlighting a stronger intention-behaviour relationship for girls. CONCLUSIONS: Findings confirm that the TPB is a successful model for predicting breakfast intentions and behaviours in adolescents. The potential for a direct effect of attitudes on behaviours should be considered in the implementation and design of breakfast interventions.

Flavonoid rich dark cocoa may improve fatigue in people with multiple sclerosis, yet has no effect on glycaemic response: An exploratory trial.

CONTEXT: Current research suggests that dark cocoa may reduce fatigue; however, the effect on fatigue in people with MS (pwMS) has never been established. The objective of this feasibility study was to explore the acute effect of high flavonoid cocoa on measures of fatigue and glycaemic response. METHODS: This was a randomised crossover participant blind exploratory study in 12 participants (2 male and 10 female) with MS-related fatigue (>4 on the Fatigue Severity Scale; FSS). After fasting overnight, participants consumed the high flavonoid cocoa drink (350 mg gallic acid equivalents {GAE}/g) or a low flavonoid cocoa control (120 mg GAE/g), consuming the alternative drink on the next visit. Fatigue was self-reported on a 100 mm visual analogue scale at 30-min time intervals for 2 h post cocoa consumption and every 2 h for the rest of the day. Fatigability was monitored using a 6 min walk test (6MWT) at the end of the visit (2 h), and activity monitors worn for 24 h commencing at 12 noon on the day of testing. The feasibility of performing the trial including outcome measures was documented. RESULTS: A moderate effect was found in self-reported fatigue throughout the day in favour of the high flavonoid group (Cohen's d 0.32, 95% non-central t CI -0.57 to 1.20). Fatigability measures did not change. Participants consumed and enjoyed the cocoa, all participants completed the study and outcome measures were accepted. CONCLUSION: The results of this study support further trials to investigate the feasibility and efficacy of pure cocoa as a dietary supplement for fatigue in pwMS.

Diamond-coated 'black silicon' as a promising material for high-surface-area electrochemical electrodes and antibacterial surfaces.

This report describes a method to fabricate high-surface-area boron-doped diamond (BDD) electrodes using so-called 'black silicon' (bSi) as a substrate. This is a synthetic nanostructured material that contains high-aspect-ratio nano-protrusions, such as spikes or needles, on the Si surface produced via plasma etching. We now show that coating a bSi surface composed of 15 µm-high needles conformably with BDD produces a robust electrochemical electrode with high sensitivity and high electroactive area. A clinically relevant demonstration of the efficacy of these electrodes is shown by measuring their sensitivity for detection of dopamine (DA) in the presence of an excess of uric acid (UA). Finally, the nanostructured surface of bSi has recently been found to generate a mechanical bactericidal effect, killing both Gram-negative and Gram-positive bacteria at high rates. We will show that BDD-coated bSi also acts as an effective antibacterial surface, with the added advantage that being diamond-coated it is far more robust and less likely to become damaged than Si.

Oral versus intubated feeding and the effect on glycaemic and insulinaemic responses, gastric emptying and satiety.

Cephalic phase responses (CPR) are important in early initiation of digestion and maximal absorption of nutrients prior to ingestion. Bypassing CPR has been shown to have consequences on metabolic responses that may influence satiety. The aim of this study was to investigate if using gastric intubation to bypass oro-pharyngeal and oesophageal exposure would reduce CPR including insulin and blood glucose and whether these impact on gastric emptying and satiety. Ten male subjects were tested on 2 occasions, 3-7 days apart after an overnight fast, in randomized order. Subjects were cannulated and intubated with a gastric tube for both tests. For test one, subjects ate 400 ml soup with a spoon and for test two the soup was infused into the stomach at an equivalent rate. Subsequently measurements of glycaemic (GR) and insulinaemic responses (IR) from cannula samples, breath samples for measurement of gastric emptying using the [(13)C] sodium acetate breath test and visual analogue scales (VAS) for satiety were taken over 180 min. There were differences in IR over the first 15 min (Oral: 169.0 ± 22.1; Gastric 124.1 ± 18.8; t(9) = 2.67; p = 0.028) but no difference in GR. There were differences in gastric emptying half time (Oral: 85.0 ± 2.7; Gastric 79.4 ± 3.3; t(9) = 2.40; p = 0.04) and ascension time (Oral: 68.2 ± 2.2; Gastric 64.0 ± 2.2; t(9) = 2.57; p = 0.03) with food taking longer to empty from the stomach on the Oral test day than on the Gastric test day. There was no significant difference in the satiety ratings. This study demonstrated that bypassing oro-pharyngeal and oesophageal exposure decreases the normal physiological CPR with detriment to IR and gastric emptying.

A comparison of body composition measurement techniques.

BACKGROUND: An understanding of the reproducibility of body composition measurements is essential for effective intervention studies. Air displacement plethysmography (ADP) and bioelectrical impedance (BIA) are two easy-to-use measures of body composition. The present study aimed to assess the reproducibility of ADP and BIA and compare them with each other, as well as with skinfold measurement. METHODS: Forty-one participants were tested on two occasions following an overnight fast. On test day 1, participants' height, weight and % body fat (BF) were measured using ADP and BIA. Measurements were replicated to assess the within-day reproducibility. On test day 2, participants were again tested using ADP and BIA and had skinfold measurements taken. Three skinfold equations for BF calculation were applied. Comparisons of within- and between-day reproducibility and between measurement techniques were completed using Pearson correlations and Bland-Altman analysis. RESULTS: Both Pearson correlation and Bland-Altman analysis showed good within- and between-day relationships and agreement for BF from ADP and BIA measurements. The two methods had a high correlation between them; however, the mean difference between the two was 3.1% (4.1%). From the skinfold equations used, the best agreement with ADP had a mean difference of 0.3% (0.8%) and, with BIA, had mean differences of 1.9% (4.2%). CONCLUSIONS: The data indicate that ADP and BIA cannot be used interchangeably, although both measurements had good within- and between-day agreement.

Comparison of the GEM and the ECAL indirect calorimeters against the Deltatrac for measures of RMR and diet-induced thermogenesis.

The Deltatrac™ II Metabolic Monitor (Datex-Ohmeda Inc.) is considered the standard reference machine in indirect calorimetry; however, it is no longer commercially available thus there is a need for new machines. The gas exchange measurement (GEM; GEM Nutrition Ltd) and the ECAL (Health Professional Solutions) are alternative measuring systems. The aim of this study was to compare the ECAL and GEM with Deltatrac for measures of RMR and the GEM to the Deltatrac for measures of diet-induced thermogenesis (DIT). Twenty healthy participants were tested on test day 1 (T1) and test day 2 (T2). RMR was measured in a randomised order for 30 min on the Deltatrac, the GEM and the ECAL. Following this, a 1553 kJ meal was consumed and DIT was measured on the Deltatrac and the GEM in alternating 15 min intervals for 4 h. The GEM reported consistently higher values than the Deltatrac for VO2, VCO2, RMR and fat oxidation (P < 0·005). The ECAL was significantly higher than the Deltatrac for measures of VO2, RMR, carbohydrate oxidation (T2) and respiratory quotient and fat oxidation (T1, T2) (P < 0·05). There were no significant differences within repeated RMR measures on the ECAL, the GEM or the Deltatrac. DIT measures were consistently higher on the GEM (T1) (P < 0·005); however, there were no significant differences between repeated measures. The findings suggest that while the GEM and the ECAL were not accurate alternatives to the Deltatrac, they may be reliable for repeated measures.

Molecular weight of barley ß-glucan influences energy expenditure, gastric emptying and glycaemic response in human subjects.

Barley ß-glucan (BG) has been shown to reduce glycaemic response (GR) in some studies. It is hypothesised that this reduction may be a function of its physical properties that delay gastric emptying (GE). The effect of these changes in GR and GE on diet-induced thermogenesis (DIT) is not known. The aim of the present study was to assess the effect of BG of different molecular weights and purities on GR, GE and DIT in healthy subjects. This was a randomised, single-blind, repeated-measures design where fifteen healthy subjects were tested on three occasions following an overnight fast. Following the baseline measurements, the volunteers were fed a soup containing high-molecular-weight BG (HBG), a soup containing low-molecular-weight BG (LBG) or a control soup with no BG (CHO). Following the consumption of the breakfast, GR was measured using finger-prick blood samples, GE was determined using the 13C-octanoic acid breath test and DIT was measured using indirect calorimetry. There was a difference in GR AUC between the soups after 60 min but not after 120 min. The CHO and LBG meals had a greater GR than the HBG meal. There were differences in all GE time points, with the HBG meal having the slowest GE time. There was a correlation between the GR and the initial GE times. There were differences in total DIT between the three test meals with the HBG meal having the lowest DIT. The present study indicates that HBG has the ability to delay GE due to increased viscosity, resulting in a decreased GR and DIT.

Soups increase satiety through delayed gastric emptying yet increased glycaemic response.

BACKGROUND/OBJECTIVES: Previous studies have demonstrated the satiating properties of soups compared with solids; however, the mechanisms controlling soup-induced satiety are unknown. This study aimed to understand the physiological mechanisms causing soup to be more satiating. SUBJECTS/METHODS: A total of 12 volunteers were tested on three occasions after a solid meal, chunky soup or smooth soup test meal for gastric emptying (GE) using the sodium [1-¹³C] acetate breath test, satiety using visual analog scales (VAS) and glycaemic response (GR) using finger prick blood samples. RESULTS: There was a significant difference in GE half-time (P=0.022) and GE ascension time (P=0.018), with the longest GE times for the smooth soup and the shortest for the solid meal. The GR area under the curve was significantly different between meals (P=0.040). The smooth soup had the greatest GR (87.0 ± 49.5 mmol/l/min), followed by the chunky soup (65.4 ± 48.0 mmol/l/min), with the solid meal having the lowest GR (61.6 ± 36.8 mmol/l/min). Volunteers were fuller after the smooth soup compared with solid meal (P=0.034). CONCLUSIONS: The smooth soup induced greater fullness compared with the solid meal because of a combination of delayed GE leading to feelings of gastric distension and rapid accessibility of nutrients causing a greater glycaemic response.

Gastrointestinal transit, post-prandial lipaemia and satiety following 3 days high-fat diet in men.

BACKGROUND/OBJECTIVES: High-fat (HF) diets of 2 weeks have been shown to accelerate gastric emptying (GE). To date, no studies have shown any alteration in GE following shorter HF diets. The aim of this study was to assess if an HF, high-energy diet of 3 days can adapt gastrointestinal (GI) transit, blood lipids and satiety. SUBJECTS/METHODS: Eleven male volunteers participated in a study consisting of three, 3-day interventions each separated by a test day. During the first intervention, volunteers recorded their diet. In the second and third interventions, volunteers repeated their food diary plus either a low-fat yogurt or HF yogurt supplement in randomized order. Test days involved measurement of GE using the (13)C octanoic-acid breath-test, mouth-to-caecum transit time (MCTT) using the inulin H(2) breath test and satiety using visual analogue scales. Blood samples for measurement of lipaemia were taken using a venous cannula. RESULTS: MCTT was different between the three test days (P=0.038), with the shortest MCTT following the HF intervention. GE was shortest following the HF intervention. There were no differences in satiety between the interventions. The HF intervention reduced triglycerides, total cholesterol and low-density lipoprotein cholesterol, and increased high-density lipoprotein cholesterol. CONCLUSION: This study shows that changes in GI transit owing to an HF diet can occur in a time period as short as 3 days.

Genome comparisons reveal a dominant mechanism of chromosome number reduction in grasses and accelerated genome evolution in Triticeae.

Single-nucleotide polymorphism was used in the construction of an expressed sequence tag map of Aegilops tauschii, the diploid source of the wheat D genome. Comparisons of the map with the rice and sorghum genome sequences revealed 50 inversions and translocations; 2, 8, and 40 were assigned respectively to the rice, sorghum, and Ae. tauschii lineages, showing greatly accelerated genome evolution in the large Triticeae genomes. The reduction of the basic chromosome number from 12 to 7 in the Triticeae has taken place by a process during which an entire chromosome is inserted by its telomeres into a break in the centromeric region of another chromosome. The original centromere-telomere polarity of the chromosome arms is maintained in the new chromosome. An intrachromosomal telomere-telomere fusion resulting in a pericentric translocation of a chromosome segment or an entire arm accompanied or preceded the chromosome insertion in some instances. Insertional dysploidy has been recorded in three grass subfamilies and appears to be the dominant mechanism of basic chromosome number reduction in grasses. A total of 64% and 66% of Ae. tauschii genes were syntenic with sorghum and rice genes, respectively. Synteny was reduced in the vicinity of the termini of modern Ae. tauschii chromosomes but not in the vicinity of the ancient termini embedded in the Ae. tauschii chromosomes, suggesting that the dependence of synteny erosion on gene location along the centromere-telomere axis either evolved recently in the Triticeae phylogenetic lineage or its evolution was recently accelerated.

The effect of exposure to irrigant solutions on apical dentin biofilms in vitro.

This study assessed the effectiveness of different concentrations of sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX) (Vista Dental Products, Racine, WI), and BioPure MTAD (Dentsply Endodontics-Tulsa Dental, Tulsa, OK). Intracanal contents were collected from 10 patients diagnosed with chronic apical periodontitis. The samples were cultured on hemisections of root apices to generate a polymicrobial biofilm. Each biofilm was separately immersed in 6% NaOCl, 3% NaOCl, 1% NaOCl, 2% CHX, 1% NaOCl followed by BioPure MTAD, and sterile phosphate buffered solution (PBS). SEM analysis showed 6% NaOCl and 3% NaOCl were capable of disrupting and removing the biofilm; 1% NaOCl and 1% NaOCl followed by MTAD were capable of disrupting the biofilm, but not eliminating bacteria; 2% CHX was not capable of disrupting the biofilm. Viable bacteria could not be cultured from specimens exposed to 6% NaOCl, 2 % CHX, or 1% NaOCl followed by BioPure MTAD. These results indicate that 6% NaOCl was the only irrigant capable of both rendering bacteria nonviable and physically removing the biofilm.

Cardiac abnormalities induced by zinc deficiency are associated with alterations in the expression of genes regulated by the zinc-finger transcription factor GATA-4.

Zinc (Zn) deficiency during pregnancy results in a wide variety of developmental abnormalities. The objective of this study was to determine if expression of cardiac developmental genes regulated by Zn-finger transcription factors could be modulated during dietary Zn deficiency. Rats were fed 0.5 (low Zn) or 90 (controls) microg Zn/g diet throughout pregnancy. Fetal development was examined and RNA isolated at gestation day (GD) 13 and 20. Cardiac abnormalities were detected at GD 20 in 82% of fetuses from dams fed low Zn diets compared with only 2% in controls. Cardiac developmental gene expression regulated by the Zn-finger transcription factor, GATA-4, was measured by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). In GD 13 and 20 hearts, two genes critical for heart development, alpha-myosin heavy chain (alpha-MHC) and cardiac troponin I (cTnI), were down-regulated in Zn-deficient fetuses. Expression of alpha-MHC was 66 and 40% lower at GD 13 and 20, respectively, in fetuses from dams fed low Zn diets compared with fetuses from control dams (p<0.05). Fetal cardiac TnI RNA levels were reduced 40 and 45% at GD 13 and 20 in the Zn-deficient group compared with controls (p<0.05). Fetal cardiac transcript levels of GATA-4 and MHox, a gene regulated by a helix-loop-helix transcription factor, whose expressions are not Zn-dependent, were unaffected by diet. These data indicated that alterations in gene regulation might be an underlying mechanism of cardiac abnormalities. Dysfunction of other Zn-dependent transcription factors may be an integral part of the extensive teratogenesis associated with Zn deficiency.

Microsatellite markers in avocado (Persea americana Mill.): genealogical relationships among cultivated avocado genotypes.

Twenty-five microsatellite markers uniquely differentiated 35 avocado cultivars and two wild relatives. Average heterozygosity was high (60.7%), ranging from 32% in P. steyermarkii to 84% in Fuerte and Bacon. In a subset of 15 cultivars, heterozygosity averaged 63.5% for microsatellites, compared to 41.8% for restriction fragment length polymorphisms (RFLPs). A neighbor-joining tree, according to average shared allele distances, consisted of three clusters likely corresponding to the botanical races of avocado and intermediate clusters uniting genotypes of presumably racially hybrid origin. Several results were at odds with existing botanical assignments that are sometimes rendered difficult by incomplete pedigree information, the complexity of the hybrid status (multiple backcrossing), or both. For example, cv. Harvest clustered with the Guatemalan race cultivars, yet it is derived from the Guatemalan x Mexican hybrid cv. Gwen. Persea schiedeana grouped with cv. Bacon. The rootstock G875 emerged as the most divergent genotype in our data set. Considerable diversity was found particularly among accessions from Guatemala, including G810 (West Indian race), G6 (Mexican race), G755A (hybrid Guatemalan x P. schiedeana), and G875 (probably not P. americana). Low bootstrap support, even upon exclusion of (known) hybrid genotypes from the data matrix, suggests the existence of ancient hybridization or that the botanical races originated more recently than previously thought.

Implant surface roughness affects osteoblast gene expression.

The transcription factor Cbfa1 regulates osteoblast differentiation and expression of genes necessary for the development of a mineralized phenotype. The purpose of this study was to determine if Cbfa1 and BSPII gene expression are influenced by implant surface microtopography. Osteoblasts were cultured on 600-grit (grooved) or sandblasted (roughened) cpTi implant discs. Mineralization was evaluated by Alizarin-Red-S staining. Real Time PCR was used for quantitative analysis of Cbfa1 and BSPII gene expression. Enhanced mineralization was seen in osteoblasts grown on roughened implant surfaces relative to tissue culture plastic. Real Time PCR showed significant (P < 0.05) increases in Cbfa1 gene expression in cells grown on roughened, as compared with grooved, implant surfaces. BSPII gene expression was also increased on rough surfaces in the UMR cells, but was reduced in the rat calvarial osteoblast cultures. These results suggest that osteoblast gene expression and mineralization are affected by roughened implant surface microtopographies during osseointegration of dental implants.

A decrease in intracellular zinc level precedes the detection of early indicators of apoptosis in HL-60 cells.

Low extracellular zinc concentrations have been associated with the induction of apoptosis. To assess the relationship between intracellular zinc concentration and the rate of apoptosis, cells were grown in media containing 0.5, 25, or 50 microM zinc and analyzed by flow cytometry or fluorescence microscopy. Cells grown in 0.5 microM zinc medium over 48 h showed a successive decrease in intracellular zinc concentration measured by the zinc-specific fluorophore, zinquin. After 18 h in 0.5 microM zinc medium, rhodamine 123 retention decreased. However, the addition of 10 microM zinc to the 0.5 microM medium before 16 h in culture restored rhodamine retention in the cells. After 30 h there was an increase in the number of cells cultured in 0.5 microM zinc medium that bound annexin V-FITC. These data indicated that decreased intracellular zinc concentration preceded early markers of apoptosis, with alterations in mitochondrial transmembrane potential preceding the loss of polarity in the cell membrane.

Dynamics of mobile element activity in chalcone synthase loci in the common morning glory (Ipomoea purpurea).

Mobile element dynamics in seven alleles of the chalcone synthase D locus (CHS-D) of the common morning glory (Ipomoea purpurea) are analyzed in the context of synonymous nucleotide sequence distances for CHS-D exons. By using a nucleotide sequence of CHS-D from the sister species Ipomoea nil (Japanese morning glory [Johzuka-Hisatomi, Y., Hoshino, A., Mori, T., Habu, Y. & Iida, S. (1999) Genes Genet. Syst. 74, 141-147], it is also possible to determine the relative frequency of insertion and loss of elements within the CHS-D locus between these two species. At least four different types of transposable elements exist upstream of the coding region, or within the single intron of the CHS-D locus in I. purpurea. There are three distinct families of miniature inverted-repeat transposable elements (MITES), and some recent transpositions of Activator/Dissociation (Ac/Ds)-like elements (Tip100), of some short interspersed repetitive elements (SINEs), and of an insertion sequence (InsIpCHSD) found in the neighborhood of this locus. The data provide no compelling evidence of the transposition of the mites since the separation of I. nil and I. purpurea roughly 8 million years ago. Finally, it is shown that the number and frequency of mobile elements are highly heterogeneous among different duplicate CHS loci, suggesting that the dynamics observed at CHS-D are locus-specific.