Two-billion-year-old evaporites capture Earth's great oxidation.

Major changes in atmospheric and ocean chemistry occurred in the Paleoproterozoic era (2.5 to 1.6 billion years ago). Increasing oxidation dramatically changed Earth's surface, but few quantitative constraints exist on this important transition. This study describes the sedimentology, mineralogy, and geochemistry of a 2-billion-year-old, ~800-meter-thick evaporite succession from the Onega Basin in Russian Karelia. The deposit consists of a basal unit dominated by halite (~100 meters) followed by units dominated by anhydrite-magnesite (~500 meters) and dolomite-magnesite (~200 meters). The evaporite minerals robustly constrain marine sulfate concentrations to at least 10 millimoles per kilogram of water, representing an oxidant reservoir equivalent to more than 20% of the modern ocean-atmosphere oxidizing capacity. These results show that substantial amounts of surface oxidant accumulated during this critical transition in Earth's oxygenation.

Is regular exercise an effective strategy for weight loss maintenance?

Weight regain after weight loss is one of the most significant challenges to successful obesity treatment. Regular exercise has long been touted as a strategy for weight loss maintenance, but the lack of clear evidence in clinical trials has caused some to question its effectiveness. In this review, we present the arguments both questioning and in support of exercise as an obesity therapeutic. Our purpose is to bring clarity to the literature, present a unified perspective, and identify the gaps in knowledge that need to be addressed in future studies. Critical questions remain including sex differences, individual variability and compensatory behaviors in response to exercise, exercise adherence, the role of energy flux and the molecular mechanisms mediating the beneficial effects of exercise after weight loss and during weight regain. Future research should focus on these critical questions to provide a more complete understanding of the potential benefits of exercise on weight loss maintenance.

A Pleistocene ice core record of atmospheric O2 concentrations.

The history of atmospheric O2 partial pressures (Po2) is inextricably linked to the coevolution of life and Earth's biogeochemical cycles. Reconstructions of past Po2 rely on models and proxies but often markedly disagree. We present a record of Po2 reconstructed using O2/N2 ratios from ancient air trapped in ice. This record indicates that Po2 declined by 7 per mil (0.7%) over the past 800,000 years, requiring that O2 sinks were ~2% larger than sources. This decline is consistent with changes in burial and weathering fluxes of organic carbon and pyrite driven by either Neogene cooling or increasing Pleistocene erosion rates. The 800,000-year record of steady average carbon dioxide partial pressures (Pco2) but declining Po2 provides distinctive evidence that a silicate weathering feedback stabilizes Pco2 on million-year time scales.

The role for adipose tissue in weight regain after weight loss.

Weight regain after weight loss is a substantial challenge in obesity therapeutics. Dieting leads to significant adaptations in the homeostatic system that controls body weight, which promotes overeating and the relapse to obesity. In this review, we focus specifically on the adaptations in white adipose tissues that contribute to the biological drive to regain weight after weight loss. Weight loss leads to a reduction in size of adipocytes and this decline in size alters their metabolic and inflammatory characteristics in a manner that facilitates the clearance and storage of ingested energy. We present the hypothesis whereby the long-term signals reflecting stored energy and short-term signals reflecting nutrient availability are derived from the cellularity characteristics of adipose tissues. These signals are received and integrated in the hypothalamus and hindbrain and an energy gap between appetite and metabolic requirements emerges and promotes a positive energy imbalance and weight regain. In this paradigm, the cellularity and metabolic characteristics of adipose tissues after energy-restricted weight loss could explain the persistence of a biological drive to regain weight during both weight maintenance and the dynamic period of weight regain.

Validation of photographic food records in children: are pictures really worth a thousand words?

BACKGROUND/OBJECTIVES: Self-reported food records are commonly used to estimate dietary intake. However, diet diaries are time consuming for participants and children are often unfamiliar with standard portion sizes or weights/volume of foods that can add to the error associated with self-reported intake. We hypothesize that photographic food records to assess dietary intake will be as accurate as a standard food diary and will decrease participant/family burden. SUBJECTS/METHODS: A total of 28 healthy subjects, 10-16 years, consumed a weighed diet for 3 days and returned any uneaten items for weigh back on day 4. During the 3 days of weighed diet, subjects recorded all intake both using a standard diet diary and taking photographs before and after consumption. Photographs were analyzed by two independent dieticians for estimation of serving size. The actual amount consumed was compared to the diary and photographic estimates through Spearman's correlation coefficients and confidence intervals. RESULTS: There was no difference between the diet diary and photographic estimates of total energy, carbohydrate, fat, protein, fiber, vitamins A, D and E, calcium, iron or zinc compared to actual intake. However, both participants and their parents reported that the photographic method was quicker, simpler and would be preferred if they were to record dietary intake in the future. In this study cohort, 36% of subjects accurately reported actual daily energy intake (+/-5% of actual intake), only 29% underreported energy intake and 35% overreported energy intake. CONCLUSIONS: Photographic food records can be used to accurately estimate dietary intake in a pediatric population. In addition, this method is less burdensome for the participants and their family.

Quantitative real-time reverse transcription-PCR analysis of deformed wing virus infection in the honeybee (Apis mellifera L.).

Deformed wing virus (DWV) can cause wing deformity and premature death in adult honeybees, although like many other bee viruses, DWV generally persists as a latent infection with no apparent symptoms. Using reverse transcription (RT)-PCR and Southern hybridization, we detected DWV in all life stages of honeybees, including adults with and without deformed wings. We also found DWV in the parasitic mite Varroa destructor, suggesting that this mite may be involved in the transmission of DWV. However, the detection of the virus in life stages not normally associated with mite parasitism (i.e., eggs and larvae) suggests that there are other modes of transmission. The levels of DWV in different life stages of bees were investigated by using TaqMan real-time quantitative RT-PCR. The amounts of virus varied significantly in these different stages, and the highest levels occurred in pupae and in adult worker bees with deformed wings. The variability in virus titer may reflect the different abilities of bees to resist DWV infection and replication. The epidemiology of DWV is discussed, and factors such as mite infestation, malnutrition, and climate are also considered.

Characterization and potential use of a Cryptosporidium parvum virus (CPV) antigen for detecting C. parvum oocysts.

The purpose of this study was to characterize the viral symbiont (CPV) of Cryptosporidium parvum sporozoites and evaluate the CPV capsid protein (CPV40) as a target for sensitive detection of the parasite. Recombinant CPV40 was produced in Escherichia coli, purified by affinity chromatography, and used to prepare polyclonal rabbit sera specific for the viral capsid protein. Anti-rCPV40 recognized a 40 kDa and a 30 kDa protein in C. parvum oocysts and appeared to localize to the apical end of the parasite. Anti-rCPV40 serum was capable of detecting as few as 1 C. parvum oocyst in a dot blot assay, the sensitivity being at least 1000-fold greater than sera reactive with total native C. parvum oocyst protein or specific for the 41 kDa oocyst surface antigen. Water samples were seeded with C. parvum oocysts and incubated at 4, 20, or 25 degrees C for greater than 3 months to determine if CPV levels were correlated with oocyst infectivity. Samples were removed monthly and subjected to mouse and cell culture infectivity, as well as PCR analysis for infectivity and viral particle presence. While sporozoite infectivity declined by more than 75% after 1 month at 25 degrees C, the CPV signal was similar to that of control samples at 4 degrees C. By 3 months at 20 degrees C, the C. parvum oocysts were found to be non-infectious, but retained a high CPV signal. This study indicates that CPV is an excellent target for sensitive detection of C. parvum oocysts in water, but may persist for an indefinite time after oocysts become non-infectious.

Morphological, Molecular, and Differential-Host Characterization of Meloidogyne floridensis n. sp. (Nematoda: Meloidogynidae), a Root-Knot Nematode Parasitizing Peach in Florida.

A root-knot nematode, Meloidogyne floridensis n. sp., is described and illustrated from peach originally collected from Gainesville, Florida. This new species resembles M. incognita, M. christiei, M. graminicola, and M. hispanica, but with LM and SEM observations it differs from these species either by the body length, shape of head, tail and tail terminus of second-stage juveniles, body length and shape of spicules in males, and its distinctive female perineal pattern. This pattern has a high to narrowly rounded arch with coarsely broken and network-like striae in and around anal area, faint lateral lines interrupting transverse striae, a sunken vulva and anus, and large distinct phasmids. Molecular data from ribosomal IGS illustrate that M. floridensis n. sp. is different from the mitotic species M. arenaria, M. incognita, and M. javanica. Data from RAPDs confirm it and suggest that this new species lies in an intermediate phylogenetic position between the previous species and the meiotic species M. hapla, M. fallax, and M. chitwoodi. Differential host tests based on annual crops and on Prunus accessions are reported.

Quantitation of a Glyptapanteles indiensis polydnavirus gene expressed in parasitized host, Lymantria dispar, by real-time quantitative RT-PCR.

Glyptapanteles indiensis is a polydnavirus-carrying wasp that parasitizes early instar gypsy moth larvae. During oviposition, the wasp injects calyx fluid containing polydnavirus along with its eggs into the host. Within the host, expression of polydnavirus genes triggers a set of changes in host physiology, which are of critical importance for the survival of the wasp. In the present study, a G. indiensis polydnavirus (GiPDV) gene, represented by cDNA clone GiPDV 1.1, was selected for expression analysis in the parasitized host. The GiPDV 1.1 gene transcript was detected in host hemolymph 30 min post-parasitization (pp) and continued to be detected for six days. The level of GiPDV 1.1 expression varied in different host tissues and expression in the brain was lower than in the hemolymph. The findings suggest that GiPDV 1.1 could be involved in early protection of parasitoid eggs from host cellular encapsulation. The temporal and spatial variations in PDV gene expression in different host tissues post-parasitization affirm their specific host regulation mechanism.

The future of the carbon cycle: review, calcification response, ballast and feedback on atmospheric CO2.

The operation of the carbon cycle forms an important part of the processes relevant to future changes in atmospheric carbon dioxide. The balance of carbon between terrestrial and oceanic reservoirs is an important factor and here we focus in particular on the oceans. Future changes in the carbon cycle that may affect air-sea partitioning of CO(2) are difficult to quantify but the palaeoceanographic record and modern observational studies provide important evidence of what variations might occur. These include changes in surface nutrient use, the oceanic inventory of nutrients, and the elemental composition and rain-rate ratio of marine particles. Recent work has identified two inter-linked processes of potential importance that we consider in some detail: the response of marine calcification to changes in surface water CO(2) and the association of particulate organic carbon with ballast minerals, in particular biogenic calcite. We review evidence from corals, coccolithophores and foraminifera, which suggests that the response of reduced calcification provides a negative feedback on rising atmospheric CO(2). We then use a box model to demonstrate how the calcification response may affect the organic carbon rain rate through the ballast effect. The ballast effect on export fluxes of organic and inorganic carbon acts to counteract the negative calcification response to increased CO(2). Thus, two oceanic buffers exert a significant control on ocean-atmosphere carbonate chemistry: the thermodynamic CO(2) buffer; and the ballast/calcification buffer. Just how tightly coupled the rain-rate ratio of CaCO(3)/C(org) is to fluxes of ballast minerals is an important question for future research.

Folate protects against oxidative modification of human LDL.

Elevated plasma total homocysteine is considered to be a graded risk factor for cardiovascular disease. Folate, through its homocysteine-lowering potential, may therefore be protective. Folate, however, may have protective effects independent of homocysteine-lowering. We have measured the effects of folate on Cu-catalysed oxidative damage to the unsaturated lipids in human LDL. Experiments were carried out in the presence of citrate, and followed increases in absorption at 234 nm, which measures the amount of conjugated diene produced. There is a lag time during which endogenous antioxidants are oxidised, followed by rapid oxidation of lipid. Addition of 0-6 microm - 5-methyltetrahydrofolate produced a dose-dependent increase in the lag time, suggesting that folate may have a direct anti-oxidant role in vivo, which is independent of any indirect effects through lowering of homocysteine levels.

Postnatal development of hepatocellular apolipoprotein B assembly and secretion in the rat.

In this study, we explored the paradox that in suckling rats the serum concentration of LDL is high although the liver secretes only minimal quantities of VLDL, the presumed precursor of LDL. Freshly isolated hepatocytes and hepatocytes in primary culture obtained from adult (90 days old) and suckling (17 days old) rats were used to investigate the synthesis and secretion of apolipoprotein B (apoB) and lipids as well as the density profile of secreted apoB-containing lipoproteins. Furthermore, the effects of dexamethasone and oleate on apoB biogenesis were investigated in primary cultures of hepatocytes from adult and suckling rats. Hepatocytes from suckling rats were unable to assemble mature VLDL but secreted apoB as primordial lipoprotein particles in the LDL-HDL density range. Intracellular degradation of apoB was also reduced in hepatocytes from suckling rats compared with that in hepatocytes from adults. The immaturity in VLDL assembly and apoB degradation of hepatocytes from suckling rats could be overcome by treating the cultures with dexamethasone plus oleate or dexamethasone alone. The lower microsomal triacylglycerol transfer protein (MTP) mRNA concentrations in hepatocytes from suckling rats in comparison with hepatocytes from adult rats were not reflected in lower MTP activity levels. Furthermore, dexamethasone plus oleate treatment had no effect on MTP activity although VLDL assembly and secretion were clearly stimulated. We conclude that, during the suckling period of the rat, serum LDL is directly produced by the liver. This is a result of impaired hepatic VLDL assembly, which is a consequence of low triglyceride synthesis and an inefficient mobilization of bulk lipids in the second step of VLDL assembly.

Real-time PCR for the detection of Cryptosporidium parvum.

Real time, TaqMan PCR assays were developed for the Cp11 and 18S rRNA genes of the protozoan parasite Cryptosporidium parvum. The TaqMan probes were specific for the genus Cryptosporidium, but could not hybridize exclusively with human-infectious C. parvum species and genotypes. In conjunction with development of the TaqMan assays, two commercial kits, the Mo Bio UltraClean Soil DNA kit, and the Qiagen QIAamp DNA Stool kit, were evaluated for DNA extraction from calf diarrhea and manure, and potassium dichromate and formalin preserved human feces. Real-time quantitation was achieved with the diarrhea samples, but nested PCR was necessary to detect C. parvum DNA in manure and human feces. Ileal tissues were obtained from calves at 3, 7, and 14 days post-infection, and DNA extracted and assayed. Nested PCR detected C. parvum DNA in the 7-day post-infection sample, but neither of the other time point samples were positive. These results indicate that real-time quantitation of C. parvum DNA, extracted using the commercial kits, is feasible on diarrheic feces, with large numbers of oocysts and small concentrations of PCR inhibitor(s). For samples with few oocysts and high concentrations of PCR inhibitor(s), such as manure, nested PCR is necessary for detection.

Direct evidence for a two-step assembly of ApoB48-containing lipoproteins in the lumen of the smooth endoplasmic reticulum of rabbit enterocytes.

The aim of this study was to investigate the types and characteristics of chylomicron precursors in the lumen of the secretory compartment of rabbit enterocytes. Luminal contents were separated into density subfractions in two continuous self-generating gradients of different density profiles. In enterocytes from rabbits fed a low fat diet, newly synthesized and immunodetectable apoB48 was only in the subfraction of density similar to high density lipoprotein (dense particles); the luminal triacylglycerol (TAG) content was low and only in the subfraction of density similar to that of chylomicrons/very low density lipoproteins (light particles). After feeding fat, newly synthesized, and immunodetectable apoB48 was in both dense (phospholipid-rich) and light (TAG-rich) particles. Luminal TAG mass and synthesis increased after fat feeding and was only in light particles. Pulse-chase experiments showed that the luminal-radiolabeled apoB48 lost from the dense particles was recovered in the light particles and the secreted chylomicrons. All of the light particle lipids (mass and newly synthesized) co-immunoprecipitated with apoB48. However, in the dense particles, there was a preferential co-precipitation of the preexisting rather than newly synthesized phospholipid. Assembly of apoB48-containing TAG-enriched lipoproteins is therefore a two-step process. The first step produces dense apoB48 phospholipid-rich particles, which accumulate in the smooth endoplasmic reticulum lumen. In the second step, these dense particles rapidly acquire the bulk of the TAG and additional phospholipid in a single and rapid step.

Rapid extraction of DNA From Escherichia coli and Cryptosporidium parvum for use in PCR.

The Xtra Amp tube, Isocode paper, Instagene matrix, and PrepMan matrix methods were evaluated for their ability to rapidly extract PCR-quality DNAs from Escherichia coli O157:H7 and Cryptosporidium parvum. All methods provided satisfactory DNA from E. coli, and the Xtra Amp and Instagene reagents provided satisfactory DNA from C. parvum.

A role for smooth endoplasmic reticulum membrane cholesterol ester in determining the intracellular location and regulation of sterol-regulatory-element-binding protein-2.

Cellular cholesterol homoeostasis is regulated through proteolysis of the membrane-bound precursor sterol-regulatory-element-binding protein (SREBP) that releases the mature transcription factor form, which regulates gene expression. Our aim was to identify the nature and intracellular site of the putative sterol-regulatory pool which regulates SREBP proteolysis in hamster liver. Cholesterol metabolism was modulated by feeding hamsters control chow, or a cholesterol-enriched diet, or by treatment with simvastatin or with the oral acyl-CoA:cholesterol acyltransferase inhibitor C1-1011 plus cholesterol. The effects of the different treatments on SREBP activation were confirmed by determination of the mRNAs for the low-density lipoprotein receptor and hydroxymethylglutaryl-CoA (HMG-CoA) reductase and by measurement of HMG-CoA reductase activity. The endoplasmic reticulum was isolated from livers and separated into subfractions by centrifugation in self-generating iodixanol gradients. Immunodetectable SREBP-2 accumulated in the smooth endoplasmic reticulum of cholesterol-fed animals. Cholesterol ester levels of the smooth endoplasmic reticulum membrane (but not the cholesterol levels) increased after cholesterol feeding and fell after treatment with simvastatin or C1-1011. The results suggest that an increased cellular cholesterol load causes accumulation of SREBP-2 in the smooth endoplasmic reticulum and, therefore, that membrane cholesterol ester may be one signal allowing exit of the SREBP-2/SREBP-cleavage-regulating protein complex to the Golgi.

Superior role of apolipoprotein B48 over apolipoprotein B100 in chylomicron assembly and fat absorption: an investigation of apobec-1 knock-out and wild-type mice.

Editing of apolipoprotein (apo)-B100 mRNA to yield apo-B48 is a specific and developmentally regulated step in enterocytes of mammals. However, the functional significance of this step is not known. Since mice containing only apo-B100 have not been documented to exhibit any difference in intestinal fat absorption from wild-type mice, the evolutionary advantage of apoB mRNA editing has been questioned. In the present study, we have compared fat absorption and chylomicron assembly in apobec-1 knock-out (KO) or wild-type (WT) mice subjected to different dietary manipulations: low-fat chow, a fat-enriched 'Western' diet and overnight fasting. Experiments in vivo and in vitro revealed differences in the ability of KO and WT enterocytes to assemble and secrete chylomicrons under different dietary conditions. After overnight fasting, chylomicron secretion is reduced considerably in KO compared with WT enterocytes. This is not due to reduced synthesis of apo-B or triacylglycerol (TAG), but appears to be a result of impaired assembly of chylomicrons, so that triacylglycerol accumulates in the enterocytes. After feeding with fat, secretion of chylomicrons enriched in pre-existing TAG is stimulated in KO compared with WT mice. In the present study, we have documented for the first time that apo-B100 is considerably less efficient than apo-B48 in exerting its role in the early stage of chylomicron assembly, which is rate-limiting under conditions of low dietary fat. However, this impairment is overcome by increased TAG stores that stimulate later stages in assembly, which are rate-limiting in the fat-fed state. apo-B mRNA editing may result in more efficient fat absorption, specifically under conditions of food shortage or low-fat content, and thus provide an evolutionary advantage.

Separation of plasma lipoproteins in self-generated gradients of iodixanol.

The major classes of plasma lipoprotein, very low density lipoproteins (VLDL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL), are characterized on the basis of differences in density and charge ( Table 1 ). Centrifugation is the 'gold-standard' for the analysis of plasma lipo-protein classes and beta quantitation, and other analytical procedures (1-4). Lipoprotein classes are separated by flotation of plasma or serum in a series of centrifugation steps in which the density of the plasma is increased sequentially by addition of potassium bromide. Alternatively, plasma is layered beneath a discontinuous gradient and centrifuged to separate the lipoprotein classes in a single step. However, it is impractical to use these methods in a routine analytical or clinical laboratory because of the long centrifugation steps required. It is also necessary to remove the high salt concentrations used before further analysis (e.g., agarose gel electrophoresis or determination of the cholesterol and/or triglyceride levels)can be carried out. The current method used for the assay of LDL and HDL levels in the chemical pathology laboratory involves determination of total plasma cholesterol followed by selective precipitation of HDL and determination of the cholesterol remaining in the supernatant (1). From the data obtained, the LDL and HDL cholesterol levels are indirectly calculated. It is generally accepted that this method is limited and that the results are compromised by modest elevation in levels of plasma triglyceride, such as frequently occurs in clinical samples (5). Table 1 Plasma Lipoproteins Lipoprotein Protein % PL % Cho1 % TAG % Mobility % Density % Meanvalues in serum (mg/dL) Chylomicrons 2 4 5 89 Start <1.000 <10 VLDL 10 16 17 57 pre-ß <1.006 50-200 LDL 29 24 47 6 ß 1.019-1.063 200-300 HDL 49 29 19 5 α 1.063-1.210 200-300 The mean % composition of the major classis of lipoproteins is given: PL, phospholipids; chol, cholesterol and cholesterol ester; TAG, triglyceride. The mobility from the origin is pre-ß; ß; a see Fig. 1 for example. The density is the band at which the lipoproteins float from salt gradients.

Fractionation of lipoprotein subclasses in self-generated gradients of iodixanol.

Chapter 5 described the use of self-generated gradients of iodixanol for the fractionation of human plasma lipoproteins into the major classes: high-density, low-density, and very low density (HDL, LDL, and VLDL). During the metabolism of plasma HDL and LDL, the lipid and apoprotein composition of the lipoprotein particles changes in such a manner that a series of subclasses exists, each with a distinctive range of densities (1). Thus, in KBr gradients, the two major subclasses, HDL(2) and HDL(3), have densities of 1.063-1.125 g/mL and 1.125-1.21 g/mL, respectively (1). In some individuals a third subclass (HDL1) is recognized (1.055-1.063 g/mL). Electrophoretic (2) and immunological (3,4) techniques have identified additional subfractions. Likewise, subclasses of LDL have been identified and isolated using shallow KBr gradients (5,6). The major LDL subfractions are LDL(1), LDL(2), and LDL(3), which have densities of 1.025-1.034 g/mL, 1.034-1.044 g/mL, and 1.044-1.060 g/mL, respectively (6), and electrophoretic analysis has identified more subfractions (7). The subfractions of LDL are of particular interest, as the presence of small, dense LDL particles in the plasma appears to be associated with a predisposition to cardiovascular disease, and they are recognized as a major causative factor in atherosclerosis (8). Methods for monitoring the LDL subclass pattern in population studies and in dietary and drug intervention trials are thus of considerable interest. This chapter is concerned primarily with the subfractionation of LDL. Although HDL subfractionation systems using iodixanol self-generated gradients have not yet been validated by direct comparison with other methods (e.g., gradient gel electrophoresis or KBr gradient centrifugation), a protocol is included.