Isoniazid Inhibits the Heme-Based Reactivity of Mycobacterium tuberculosis Truncated Hemoglobin N

Isoniazid represents a first-line anti-tuberculosis medication in prevention and treatment. This prodrug is activated by a mycobacterial catalase-peroxidase enzyme called KatG in Mycobacterium tuberculosis), thereby inhibiting the synthesis of mycolic acid, required for the mycobacterial cell wall. Moreover, isoniazid activation by KatG produces some radical species (e.g., nitrogen monoxide), that display anti-mycobacterial activity. Remarkably, the ability of mycobacteria to persist in vivo in the presence of reactive nitrogen and oxygen species implies the presence in these bacteria of (pseudo-)enzymatic detoxification systems, including truncated hemoglobins (trHbs). Here, we report that isoniazid binds reversibly to ferric and ferrous M. tuberculosis trHb type N (or group I; Mt-trHbN(III) and Mt-trHbN(II), respectively) with a simple bimolecular process, which perturbs the heme-based spectroscopic properties. Values of thermodynamic and kinetic parameters for isoniazid binding to Mt-trHbN(III) and Mt-trHbN(II) are K = (1.1±0.1)×10⁻⁴ M, k _on = (5.3±0.6)×10³ M⁻¹ s⁻¹ and k _off = (4.6±0.5)×10⁻¹ s⁻¹; and D = (1.2±0.2)×10⁻³ M, d _on = (1.3±0.4)×10³ M⁻¹ s⁻¹, and d _off = 1.5±0.4 s⁻¹, respectively, at pH 7.0 and 20.0°C. Accordingly, isoniazid inhibits competitively azide binding to Mt-trHbN(III) and Mt-trHbN(III)-catalyzed peroxynitrite isomerization. Moreover, isoniazid inhibits Mt-trHbN(II) oxygenation and carbonylation. Although the structure of the Mt-trHbN-isoniazid complex is not available, here we show by docking simulation that isoniazid binding to the heme-Fe atom indeed may take place. These data suggest a direct role of isoniazid to impair fundamental functions of mycobacteria, e.g. scavenging of reactive nitrogen and oxygen species, and metabolism.     

Methane emissions from beef cattle grazing on semi-natural upland and improved lowland grasslands

In ruminants, methane (CH₄) is a by-product of digestion and contributes significantly to the greenhouse gas emissions attributed to agriculture. Grazed grass is a relatively cheap and nutritious feed but herbage species and nutritional quality vary between pastures, with management, land type and season all potentially impacting on animal performance and CH₄ production. The objective of this study was to evaluate performance and compare CH₄ emissions from cattle of dairy and beef origin grazing two grassland ecosystems: lowland improved grassland (LG) and upland semi-natural grassland (UG). Forty-eight spring-born beef cattle (24 Holstein–Friesian steers, 14 Charolais crossbred steers and 10 Charolais crossbred heifers of 407 (s.d. 29), 469 (s.d. 36) and 422 (s.d. 50) kg BW, respectively), were distributed across two balanced groups that grazed the UG and LG sites from 1 June to 29 September at stocking rates (number of animals per hectare) of 1.4 and 6.7, respectively. Methane emissions and feed dry matter (DM) intake were estimated by the SF₆ tracer and n-alkane techniques, respectively, and BW was recorded across three experimental periods that reflected the progression of the grazing season. Overall, cattle grazed on UG had significantly lower (P<0.001) mean daily DM intake (8.68 v. 9.55 kg/day), CH₄ emissions (176 v. 202 g/day) and BW gain (BWG; 0.73 v. 1.08 kg/day) than the cattle grazed on LG but there was no difference (P>0.05) in CH₄ emissions per unit of feed intake when expressed either on a DM basis (20.7 and 21.6 g CH₄ per kg DM intake for UG and LG, respectively) or as a percentage of the gross energy intake (6.0% v. 6.5% for UG and LG, respectively). However, cattle grazing UG had significantly (P<0.001) greater mean daily CH₄ emissions than those grazing LG when expressed relative to BWG (261 v. 197 g CH₄/kg, respectively). The greater DM intake and BWG of cattle grazing LG than UG reflected the poorer nutritive value of the UG grassland. Although absolute rates of CH₄ emissions (g/day) were lower from cattle grazing UG than LG, cattle grazing UG would be expected to take longer to reach an acceptable finishing weight, thereby potentially off-setting this apparent advantage. Methane emissions constitute an adverse environmental impact of grazing by cattle but the contribution of cattle to ecosystem management (i.e. promoting biodiversity) should also be considered when evaluating the usefulness of different breeds for grazing semi-natural or unimproved grassland.     

The effect of dietary addition of nitrate or increase in lipid concentrations,alone or in combination,on performance and methane emissions of beef cattle

Adding nitrate to or increasing the concentration of lipid in the diet are established strategies for reducing enteric methane (CH₄) emissions, but their effectiveness when used in combination has been largely unexplored. This study investigated the effect of dietary nitrate and increased lipid included alone or together on CH₄ emissions and performance traits of finishing beef cattle. The experiment was a 2×4 factorial design comprising two breeds (cross-bred Aberdeen Angus (AAx) and cross-bred Limousin (LIMx) steers) and four dietary treatments (each based on 550 g forage : 450 g concentrate/kg dry matter (DM)). The four dietary treatments were assigned according to a 2×2 factorial design where the control treatment contained rapeseed meal as the main protein source, which was replaced either with nitrate (21.5 g nitrate/kg DM); maize distillers dark grains (MDDG, which increased diet ether extract from 24 to 37 g/kg DM) or both nitrate and MDDG. Steers (n=20/dietary treatment) were allocated to each of the four treatments in equal numbers of each breed with feed offered ad libitum. After 28 days adaptation to dietary treatments, individual animal intake, performance and feed efficiency were recorded for 56 days. Thereafter, CH₄ emissions were measured over 13 weeks (six steers/week). Increasing dietary lipid did not adversely affect animal performance and showed no interactions with dietary nitrate. In contrast, addition of nitrate to diets resulted in poorer live-weight gain (P<0.01) and increased feed conversion ratio (P<0.05) compared with diets not containing nitrate. Daily CH₄ output was lower (P<0.001) on nitrate-containing diets but increasing dietary lipid resulted in only a non-significant reduction in CH₄. There were no interactions associated with CH₄ emissions between dietary nitrate and lipid. Cross-bred Aberdeen Angus steers achieved greater live-weight gains (P<0.01), but had greater DM intakes (P<0.001), greater fat depth (P<0.01) and poorer residual feed intakes (P<0.01) than LIMx steers. Cross-bred Aberdeen Angus steers had higher daily CH₄ outputs (P<0.001) but emitted less CH₄ per kilogram DM intake than LIMx steers (P<0.05). In conclusion, inclusion of nitrate reduced CH₄ emissions in growing beef cattle although the efficacy of nitrate was less than in previous work. When increased dietary lipid and nitrate inclusion were combined there was no evidence of an interaction between treatments and therefore combining different nutritional treatments to mitigate CH₄ emissions could be a useful means of achieving reductions in CH₄ while minimising any adverse effects.     

A new estimation of global soil greenhouse gas fluxes using a simple data-oriented model

Soil greenhouse gas fluxes (particularly CO₂, CH₄, and N₂O) play important roles in climate change. However, despite the importance of these soil greenhouse gases, the number of reports on global soil greenhouse gas fluxes is limited. Here, new estimates are presented for global soil CO₂ emission (total soil respiration), CH₄ uptake, and N₂O emission fluxes, using a simple data-oriented model. The estimated global fluxes for CO₂ emission, CH₄ uptake, and N₂O emission were 78 Pg C yr⁻¹ (Monte Carlo 95% confidence interval, 64–95 Pg C yr⁻¹), 18 Tg C yr⁻¹ (11–23 Tg C yr⁻¹), and 4.4 Tg N yr⁻¹ (1.4–11.1 Tg N yr⁻¹), respectively. Tropical regions were the largest contributor of all of the gases, particularly the CO₂ and N₂O fluxes. The soil CO₂ and N₂O fluxes had more pronounced seasonal patterns than the soil CH₄ flux. The collected estimates, including both the previous and the present estimates, demonstrate that the means of the best estimates from each study were 79 Pg C yr⁻¹ (291 Pg CO₂ yr⁻¹; coefficient of variation, CV = 13%, N = 6) for CO₂, 21 Tg C yr⁻¹ (29 Tg CH₄ yr⁻¹; CV = 24%, N = 24) for CH₄, and 7.8 Tg N yr⁻¹ (12.2 Tg N₂O yr⁻¹; CV = 38%, N = 11) for N₂O. For N₂O, the mean of the estimates that was calculated by excluding the earliest two estimates was 6.6 Tg N yr⁻¹ (10.4 Tg N₂O yr⁻¹; CV = 22%, N = 9). The reported estimates vary and have large degrees of uncertainty but their overall magnitudes are in general agreement. To further minimize the uncertainty of soil greenhouse gas flux estimates, it is necessary to build global databases and identify key processes in describing global soil greenhouse gas fluxes.     

The impact of divergent breed types and diets on methane emissions,rumen characteristics and performance of finishing beef cattle

This study was undertaken to further develop our understanding of the links between breed, diet and the rumen microbial community and determine their effect on production characteristics and methane (CH₄) emissions from beef cattle. The experiment was of a 2×2 factorial design, comprising two breeds (crossbred Charolais (CHX); purebred Luing (LU)) and two diets (concentrate-straw or silage-based). In total, 80 steers were used and balanced for sire within each breed, farm of origin and BW across diets. The diets (fed as total mixed rations) consisted of (g/kg dry matter (DM)) forage to concentrate ratios of either 500 : 500 (Mixed) or 79 : 921 (Concentrate). Steers were adapted to the diets over a 4-week period and performance and feed efficiency were then measured over a 56-day test period. Directly after the 56-day test, CH₄ and carbon dioxide (CO₂) emissions were measured (six steers/week) over a 13-week period. Compared with LU steers, CHX steers had greater average daily gain (ADG; P<0.05) and significantly (P<0.001) lower residual feed intake. Crossbred Charolais steers had superior conformation and fatness scores (P<0.001) than LU steers. Although steers consumed, on a DM basis, more Concentrate than Mixed diet (P<0.01), there were no differences between diets in either ADG or feed efficiency during the 56-day test. At slaughter, however, Concentrate-fed steers were heavier (P<0.05) and had greater carcass weights than Mixed-fed steers (P<0.001). Breed of steer did not influence CH₄ production, but it was substantially lower when the Concentrate rather than Mixed diet was fed (P<0.001). Rumen fluid from Concentrate-fed steers contained greater proportions of propionic acid (P<0.001) and lower proportions of acetic acid (P<0.001), fewer archaea (P<0.01) and protozoa (P=0.09), but more Clostridium Cluster XIVa (P<0.01) and Bacteroides plus Prevotella (P<0.001) than Mixed-fed steers. When the CH₄ to CO₂ molar ratio was considered as a proxy method for CH₄ production (g/kg DM intake), only weak relationships were found within diets. In conclusion, although feeding Concentrate and Mixed diets produced substantial differences in CH₄ emissions and rumen characteristics, differences in performance were influenced more markedly by breed.     

Validity of Hydration Non-Invasive Indices during the Weightcutting and Official Weigh-In for Olympic Combat Sports

Background

In Olympic combat sports, weight cutting is a common practice aimed to take advantage of competing in weight divisions below the athlete''s normal weight. Fluid and food restriction in combination with dehydration (sauna and/or exercise induced profuse sweating) are common weight cut methods. However, the resultant hypohydration could adversely affect health and performance outcomes.

Purpose

The aim of this study is to determine which of the routinely used non-invasive measures of dehydration best track urine osmolality, the gold standard non-invasive test.

Method

Immediately prior to the official weigh-in of three National Championships, the hydration status of 345 athletes of Olympic combat sports (i.e., taekwondo, boxing and wrestling) was determined using five separate techniques: i) urine osmolality (U_OSM), ii) urine specific gravity (U_SG), iii) urine color (U_COL), iv) bioelectrical impedance analysis (BIA), and v) thirst perception scale (TPS). All techniques were correlated with U_OSM divided into three groups: euhydrated (G₁; U_OSM 250–700 mOsm·kg H₂O⁻¹), dehydrated (G₂; U_OSM 701–1080 mOsm·kg H₂O⁻¹), and severely dehydrated (G₃; U_OSM 1081–1500 mOsm·kg H₂O⁻¹).

Results

We found a positive high correlation between the U_OSM and U_SG (r = 0.89: p = 0.000), although this relationship lost strength as dehydration increased (G₁ r = 0.92; G₂ r = 0.73; and G₃ r = 0.65; p = 0.000). U_COL showed a moderate although significant correlation when considering the whole sample (r = 0.743: p = 0.000) and G₁ (r = 0.702: p = 0.000) but low correlation for the two dehydrated groups (r = 0.498–0.398). TPS and BIA showed very low correlation sizes for all groups assessed.

Conclusion

In a wide range of pre-competitive hydration status (U_OSM 250–1500 mOsm·kg H₂O⁻¹), U_SG is highly associated with U_OSM while being a more affordable and easy to use technique. U_COL is a suitable tool when U_SG is not available. However, BIA or TPS are not sensitive enough to detect hypohydration at official weight-in before an Olympic combat championship.     

Physical Activity Attenuates the Genetic Predisposition to Obesity in 20,000 Men and Women from EPIC-Norfolk Prospective Population Study

Background

We have previously shown that multiple genetic loci identified by genome-wide association studies (GWAS) increase the susceptibility to obesity in a cumulative manner. It is, however, not known whether and to what extent this genetic susceptibility may be attenuated by a physically active lifestyle. We aimed to assess the influence of a physically active lifestyle on the genetic predisposition to obesity in a large population-based study.

Methods and Findings

We genotyped 12 SNPs in obesity-susceptibility loci in a population-based sample of 20,430 individuals (aged 39–79 y) from the European Prospective Investigation of Cancer (EPIC)-Norfolk cohort with an average follow-up period of 3.6 y. A genetic predisposition score was calculated for each individual by adding the body mass index (BMI)-increasing alleles across the 12 SNPs. Physical activity was assessed using a self-administered questionnaire. Linear and logistic regression models were used to examine main effects of the genetic predisposition score and its interaction with physical activity on BMI/obesity risk and BMI change over time, assuming an additive effect for each additional BMI-increasing allele carried. Each additional BMI-increasing allele was associated with 0.154 (standard error [SE] 0.012) kg/m² (p = 6.73×10⁻³⁷) increase in BMI (equivalent to 445 g in body weight for a person 1.70 m tall). This association was significantly (p _interaction = 0.005) more pronounced in inactive people (0.205 [SE 0.024] kg/m² [p = 3.62×10⁻¹⁸; 592 g in weight]) than in active people (0.131 [SE 0.014] kg/m² [p = 7.97×10⁻²¹; 379 g in weight]). Similarly, each additional BMI-increasing allele increased the risk of obesity 1.116-fold (95% confidence interval [CI] 1.093–1.139, p = 3.37×10⁻²⁶) in the whole population, but significantly (p _interaction = 0.015) more in inactive individuals (odds ratio [OR] = 1.158 [95% CI 1.118–1.199; p = 1.93×10⁻¹⁶]) than in active individuals (OR = 1.095 (95% CI 1.068–1.123; p = 1.15×10⁻¹²]). Consistent with the cross-sectional observations, physical activity modified the association between the genetic predisposition score and change in BMI during follow-up (p _interaction = 0.028).

Conclusions

Our study shows that living a physically active lifestyle is associated with a 40% reduction in the genetic predisposition to common obesity, as estimated by the number of risk alleles carried for any of the 12 recently GWAS-identified loci. Please see later in the article for the Editors'' Summary     

Priming of Soil Carbon Decomposition in Two Inner Mongolia Grassland Soils following Sheep Dung Addition: A Study Using 13C Natural Abundance Approach

To investigate the effect of sheep dung on soil carbon (C) sequestration, a 152 days incubation experiment was conducted with soils from two different Inner Mongolian grasslands, i.e. a Leymus chinensis dominated grassland representing the climax community (2.1% organic matter content) and a heavily degraded Artemisia frigida dominated community (1.3% organic matter content). Dung was collected from sheep either fed on L. chinensis (C₃ plant with δ¹³C = −26.8‰; dung δ¹³C = −26.2‰) or Cleistogenes squarrosa (C₄ plant with δ¹³C = −14.6‰; dung δ¹³C = −15.7‰). Fresh C₃ and C₄ sheep dung was mixed with the two grassland soils and incubated under controlled conditions for analysis of ¹³C-CO₂ emissions. Soil samples were taken at days 17, 43, 86, 127 and 152 after sheep dung addition to detect the δ¹³C signal in soil and dung components. Analysis revealed that 16.9% and 16.6% of the sheep dung C had decomposed, of which 3.5% and 2.8% was sequestrated in the soils of L. chinensis and A. frigida grasslands, respectively, while the remaining decomposed sheep dung was emitted as CO₂. The cumulative amounts of C respired from dung treated soils during 152 days were 7–8 times higher than in the un-amended controls. In both grassland soils, ca. 60% of the evolved CO₂ originated from the decomposing sheep dung and 40% from the native soil C. Priming effects of soil C decomposition were observed in both soils, i.e. 1.4 g and 1.6 g additional soil C kg⁻¹ dry soil had been emitted as CO₂ for the L. chinensis and A. frigida soils, respectively. Hence, the net C losses from L. chinensis and A. frigida soils were 0.6 g and 0.9 g C kg⁻¹ soil, which was 2.6% and 7.0% of the total C in L. chinensis and A. frigida grasslands soils, respectively. Our results suggest that grazing of degraded Inner Mongolian pastures may cause a net soil C loss due to the positive priming effect, thereby accelerating soil deterioration.     

Effects of Winter Cover Crops Straws Incorporation on CH4 and N2O Emission from Double-Cropping Paddy Fields in Southern China

Residue management in cropping systems is believed to improve soil quality. However, the effects of residue management on methane (CH₄) and nitrous oxide (N₂O) emissions from paddy field in Southern China have not been well researched. The emissions of CH₄ and N₂O were investigated in double cropping rice (Oryza sativa L.) systems with straw returning of different winter cover crops by using the static chamber-gas chromatography technique. A randomized block experiment with three replications was established in 2004 in Hunan Province, China, including rice–rice–ryegrass (Lolium multiflorum L.) (Ry-R-R), rice–rice–Chinese milk vetch (Astragalus sinicus L.) (Mv-R-R) and rice–rice with winter fallow (Fa-R-R). The results showed that straw returning of winter crops significantly increased the CH₄ emission during both rice growing seasons when compared with Fa-R-R. Ry-R-R plots had the largest CH₄ emissions during the early rice growing season with 14.235 and 15.906 g m⁻² in 2012 and 2013, respectively, when Ry-R-R plots had the largest CH₄ emission during the later rice growing season with 35.673 and 38.606 g m⁻² in 2012 and 2013, respectively. The Ry-R-R and Mv-R-R also had larger N₂O emissions than Fa-R-R in both rice seasons. When compared to Fa-R-R, total N₂O emissions in the early rice growing season were increased by 0.05 g m⁻² in Ry-R-R and 0.063 g m⁻² in Mv-R-R in 2012, and by 0.058 g m⁻² in Ry-R-R and 0.068 g m⁻² in Mv-R-R in 2013, respectively. Similar result were obtained in the late rice growing season, and the total N₂O emissions were increased by 0.104 g m⁻² in Ry-R-R and 0.073 g m⁻² in Mv-R-R in 2012, and by 0.108 g m⁻² in Ry-R-R and 0.076 g m⁻² in Mv-R-R in 2013, respectively. The global warming potentials (GWPs) from paddy fields were ranked as Ry-R-R>Mv-R-R>Fa-R-R. As a result, straw returning of winter cover crops has significant effects on increase of CH₄ and N₂O emission from paddy field in double cropping rice system.     

Predicting Greenhouse Gas Emissions and Soil Carbon from Changing Pasture to an Energy Crop

Bioenergy related land use change would likely alter biogeochemical cycles and global greenhouse gas budgets. Energy cane (Saccharum officinarum L.) is a sugarcane variety and an emerging biofuel feedstock for cellulosic bio-ethanol production. It has potential for high yields and can be grown on marginal land, which minimizes competition with grain and vegetable production. The DayCent biogeochemical model was parameterized to infer potential yields of energy cane and how changing land from grazed pasture to energy cane would affect greenhouse gas (CO₂, CH₄ and N₂O) fluxes and soil C pools. The model was used to simulate energy cane production on two soil types in central Florida, nutrient poor Spodosols and organic Histosols. Energy cane was productive on both soil types (yielding 46–76 Mg dry mass⋅ha⁻¹). Yields were maintained through three annual cropping cycles on Histosols but declined with each harvest on Spodosols. Overall, converting pasture to energy cane created a sink for GHGs on Spodosols and reduced the size of the GHG source on Histosols. This change was driven on both soil types by eliminating CH₄ emissions from cattle and by the large increase in C uptake by greater biomass production in energy cane relative to pasture. However, the change from pasture to energy cane caused Histosols to lose 4493 g CO₂ eq⋅m⁻² over 15 years of energy cane production. Cultivation of energy cane on former pasture on Spodosol soils in the southeast US has the potential for high biomass yield and the mitigation of GHG emissions.     

Adult-Onset Obesity Reveals Prenatal Programming of Glucose-Insulin Sensitivity in Male Sheep Nutrient Restricted during Late Gestation

Background

Obesity invokes a range of metabolic disturbances, but the transition from a poor to excessive nutritional environment may exacerbate adult metabolic dysfunction. The current study investigated global maternal nutrient restriction during early or late gestation on glucose tolerance and insulin sensitivity in the adult offspring when lean and obese.

Methods/Principal Findings

Pregnant sheep received adequate (1.0M; CE, n = 6) or energy restricted (0.7M) diet during early (1–65 days; LEE, n = 6) or late (65–128 days; LEL, n = 7) gestation (term ∼147 days). Subsequent offspring remained on pasture until 1.5 years when all received glucose and insulin tolerance tests (GTT & ITT) and body composition determination by dual energy x-ray absorptiometry (DXA). All animals were then exposed to an obesogenic environment for 6–7 months and all protocols repeated. Prenatal dietary treatment had no effect on birth weight or on metabolic endpoints when animals were ‘lean’ (1.5 years). Obesity revealed generalised metabolic ‘inflexibility’ and insulin resistance; characterised by blunted excursions of plasma NEFA and increased insulin_AUC (from 133 to 341 [s.e.d. 26] ng.ml⁻¹.120 mins) during a GTT, respectively. For LEL vs. CE, the peak in plasma insulin when obese was greater (7.8 vs. 4.7 [s.e.d. 1.1] ng.ml⁻¹) and was exacerbated by offspring sex (i.e. 9.8 vs. 4.4 [s.e.d. 1.16] ng.ml⁻¹; LEL male vs. CE male, respectively). Acquisition of obesity also significantly influenced the plasma lipid and protein profile to suggest, overall, greater net lipogenesis and reduced protein metabolism.

Conclusions

This study indicates generalised metabolic dysfunction with adult-onset obesity which also exacerbates and ‘reveals’ programming of glucose-insulin sensitivity in male offspring prenatally exposed to maternal undernutrition during late gestation. Taken together, the data suggest that metabolic function appears little compromised in young prenatally ‘programmed’ animals so long as weight is adequately controlled. Nutritional excess in adulthood exacerbates any programmed phenotype, indicating greater vigilance over weight control is required for those individuals exposed to nutritional thrift during gestation.     

Quantitative Assessment of the Association between rs2046210 at 6q25.1 and Breast Cancer Risk

Genome-wide association studies (GWAS) have identified several genetic susceptibility loci for breast cancer (BC). One of them, conducted among Chinese women, found an association of rs2046210 at 6q25.1 with the risk of BC recently. Since then, numerous association studies have been carried out to investigate the relationship between this polymorphism and BC risk in various populations. However, these have yielded contradictory results. We therefore performed a meta-analysis to clarify this inconsistency. Overall, a total of 235003 subjects based on 13 studies were included in our study. Significantly increased BC risk was detected in the pooled analysis [allele contrast: OR = 1.13, 95%CI = 1.10–1.17, P(Z) <10⁻⁵, P(Q) <10⁻⁴; dominant model: OR = 1.21, 95%CI = 1.14–1.27, P(Z) <10⁻⁵, P(Q) <10⁻⁴; recessive model: OR = 1.18, 95%CI = 1.12–1.24, P(Z) <10⁻⁵, P(Q) = 0.04]. In addition, our data revealed that rs2046210 conferred greater risk in estrogen receptor (ER)-negative tumors [OR = 1.27, 95%CI = 1.15–1.40, P(Z) <10⁻⁵, P(Q) <10⁻⁴] than in ER-positive ones [OR = 1.18, 95%CI = 1.09–1.28, P(Z) <10⁻⁴, P(Q) = 0.0003]. When stratified by ethnicity, significant associations were found in Caucasian and Asian populations, but not detected among Africans. There was evidence of heterogeneity (P<0.05), however, the heterogeneity largely disappeared after stratification by ethnicity. The present meta-analysis demonstrated that the rs2046210 polymorphism may be associated with increased BC susceptibility, but this association varies in different ethnicities.     

Transporting Antitumor Drug Tamoxifen and Its Metabolites, 4-Hydroxytamoxifen and Endoxifen by Chitosan Nanoparticles

Synthetic and natural polymers are often used as drug delivery systems in vitro and in vivo. Biodegradable chitosan of different sizes were used to encapsulate antitumor drug tamoxifen (Tam) and its metabolites 4-hydroxytamoxifen (4-Hydroxytam) and endoxifen (Endox). The interactions of tamoxifen and its metabolites with chitosan 15, 100 and 200 KD were investigated in aqueous solution, using FTIR, fluorescence spectroscopic methods and molecular modeling. The structural analysis showed that tamoxifen and its metabolites bind chitosan via both hydrophilic and hydrophobic contacts with overall binding constants of K _tam-ch-15  = 8.7 (±0.5)×10³ M⁻¹, K _tam-ch-100  = 5.9 (±0.4)×10⁵ M⁻¹, K _tam-ch-200  = 2.4 (±0.4)×10⁵ M⁻¹ and K _{hydroxytam-ch-15}  = 2.6(±0.3)×10⁴ M⁻¹, K _{hydroxytam – ch-100}  = 5.2 (±0.7)×10⁶ M⁻¹ and K _{hydroxytam-ch-200}  = 5.1 (±0.5)×10⁵ M⁻¹, K _endox-ch-15  = 4.1 (±0.4)×10³ M⁻¹, K _endox-ch-100  = 1.2 (±0.3)×10⁶ M⁻¹ and K _endox-ch-200  = 4.7 (±0.5)×10⁵ M⁻¹ with the number of drug molecules bound per chitosan (n) 2.8 to 0.5. The order of binding is ch-100>200>15 KD with stronger complexes formed with 4-hydroxytamoxifen than tamoxifen and endoxifen. The molecular modeling showed the participation of polymer charged NH₂ residues with drug OH and NH₂ groups in the drug-polymer adducts. The free binding energies of −3.46 kcal/mol for tamoxifen, −3.54 kcal/mol for 4-hydroxytamoxifen and −3.47 kcal/mol for endoxifen were estimated for these drug-polymer complexes. The results show chitosan 100 KD is stronger carrier for drug delivery than chitosan-15 and chitosan-200 KD.     

Microbial population dynamics in an extreme environment: controlling factors in talus soils at 3750 m in the Colorado Rocky Mountains

Variation of CH₄ emissions over a three-year period was studied in a reed-dominated (Phragmites australis) littoral transect of a boreal lake undergoing shoreline displacement due to postglacial rebound. The seasonal variation in plant-mediated CH₄ emissions during open-water periods was significantly correlated with sediment temperature. The highest plant-mediated emission rates (up to 2050 mg CH₄ m^–2 d^–1) were found in the outermost reed zone, where culms of the previous growing seasons had accumulated and free-floating plants grew on the decomposing culms. In reed zones closer to the shoreline as well as in mixed stands of reed and cattail, the maximum daily rates were usually > 500 mg CH₄ m^–2 d^–1. The total plant-mediated CH₄ emission during the open-water period was significantly correlated with the seasonal maximum of green shoot biomass. This relationship was strongest in the continuously flooded (water depth > 25 cm) outermost zones. In this area, emissions through ebullition were of greatest importance and could exceed plant-mediated emissions. In general, total emissions of the open-water periods varied from ca. 20 to 50 g CH₄ m^–2 a^–1, but in the outermost reed zone, the plant-mediated emissions could be as high as 123 g CH₄ m^–2 a^–1; ebullition emissions from this zone reached > 100 g CH₄ m^–2 a^–1. The proportion of CH₄ released in winter was usually < 10% of annual emissions. Emissions of CH₄ were higher in this flooded transgression shore the than those measured in boreal peatlands, but the role of ancient carbon stores as a substrate supply compared with recent anthropogenic eutrophication is unknown.     

Genetic Association Study of TNFAIP3, IFIH1, IRF5 Polymorphisms with Polymyositis/Dermatomyositis in Chinese Han Population

Background

Single-nucleotide polymorphisms (SNPs) in the TNFAIP3, IFIH1, and IRF5 genes have been associated with several auto-inflammation diseases, while the susceptibility between these genes and idiopathic inflammatory myopathies (IIMs) were not reported. This study aimed to investigate whether TNFAIP3, IFIH1, and IRF5 gene polymorphisms confer susceptibility for the IIMs in Chinese Han population.

Methods

A large case–control study of Chinese subjects with polymyositis (PM) (n = 298) and dermatomyositis (DM) (n = 530) was accomplished. 968 healthy and ethnically matched controls were available for comparison. Six SNPs in the TNFAIP3 region (rs2230926 and rs5029939), the IFIH1 gene (rs1990760 and rs3747517) and the IRF5 region (rs4728142 and rs729302) were assessed and genotyped using the Sequenom MassArray iPLEX platform.

Results

Our study indicated a strong allele association was observed in PM/DM and PM patients for rs2230926 (OR: 1.61, 95%CI: 1.20–2.16, P_c = 7.5×10⁻³; OR: 1.88, 95%CI: 1.30–2.74, P_c = 4.0×10⁻³, respectively) and rs5029939 (OR: 1.64, 95%CI: 1.21–2.21, P_c = 6.0×10⁻³; OR: 1.88, 95%CI: 1.28–2.76, P_c = 5.5×10⁻³,respectively). And rs2230926 and rs5029939 were significantly associated with interstitial lung disease (ILD) in PM/DM and PM patients (P_c = 0.04 and P_c = 0.016; P_c = 0.02 and P_c = 0.03, respectively). In addition, rs4728142 allele and genotype had significant association with PM/DM patients (P_c = 0.026 and P_c = 0.048, respectively). Further analysis with three logistic regression genetic models revealed statistically significant difference in the genotypic distribution in the PM/DM, PM or DM patients when the additive and dominant models were used.

Conclusions

This was the first study to reveal TNFAIP3 and IRF5 polymorphisms were associated with PM/DM patients or these patients with ILD, indicating that TNFAIP3 and IRF5 might be the susceptibility gene for PM/DM patients in Chinese Han population.     

Polymorphism Located between CPT1B and CHKB,and HLA-DRB1*1501-DQB1*0602 Haplotype Confer Susceptibility to CNS Hypersomnias (Essential Hypersomnia)

Background

SNP rs5770917 located between CPT1B and CHKB, and HLA-DRB1*1501-DQB1*0602 haplotype were previously identified as susceptibility loci for narcolepsy with cataplexy. This study was conducted in order to investigate whether these genetic markers are associated with Japanese CNS hypersomnias (essential hypersomnia: EHS) other than narcolepsy with cataplexy.

Principal Findings

EHS was significantly associated with SNP rs5770917 (P_allele = 3.6×10⁻³; OR = 1.56; 95% c.i.: 1.12–2.15) and HLA-DRB1*1501-DQB1*0602 haplotype (P _positivity = 9.2×10⁻¹¹; OR = 3.97; 95% c.i.: 2.55–6.19). No interaction between the two markers (SNP rs5770917 and HLA-DRB1*1501-DQB1*0602 haplotype) was observed in EHS.

Conclusion

CPT1B, CHKB and HLA are candidates for susceptibility to CNS hypersomnias (EHS), as well as narcolepsy with cataplexy.     

The Principal Genetic Determinants for Nasopharyngeal Carcinoma in China Involve the HLA Class I Antigen Recognition Groove

Nasopharyngeal carcinoma (NPC) is an epithelial malignancy facilitated by Epstein-Barr Virus infection. Here we resolve the major genetic influences for NPC incidence using a genome-wide association study (GWAS), independent cohort replication, and high-resolution molecular HLA class I gene typing including 4,055 study participants from the Guangxi Zhuang Autonomous Region and Guangdong province of southern China. We detect and replicate strong association signals involving SNPs, HLA alleles, and amino acid (aa) variants across the major histocompatibility complex-HLA-A, HLA –B, and HLA -C class I genes (P_{HLA-A-aa-site-62} = 7.4×10⁻²⁹; P _{HLA-B-aa-site-116} = 6.5×10⁻¹⁹; P _{HLA-C-aa-site-156} = 6.8×10⁻⁸ respectively). Over 250 NPC-HLA associated variants within HLA were analyzed in concert to resolve separate and largely independent HLA-A, -B, and -C gene influences. Multivariate logistical regression analysis collapsed significant associations in adjacent genes spanning 500 kb (OR2H1, GABBR1, HLA-F, and HCG9) as proxies for peptide binding motifs carried by HLA- A*11:01. A similar analysis resolved an independent association signal driven by HLA-B*13:01, B*38:02, and B*55:02 alleles together. NPC resistance alleles carrying the strongly associated amino acid variants implicate specific class I peptide recognition motifs in HLA-A and -B peptide binding groove as conferring strong genetic influence on the development of NPC in China.     

A Genome-Wide Association Study of Total Serum and Mite-Specific IgEs in Asthma Patients

Immunoglobulin E (IgE) is one of the central players in asthma and allergic diseases. Although the serum IgE level, a useful endophenotype, is generally increased in patients with asthma, genetic factors influencing IgE regulation in asthma are still not fully understood. To identify the genetic variations associated with total serum and mite-specific IgEs in asthmatics, a genome-wide association study (GWAS) of 657,366 single nucleotide polymorphisms (SNPs) was performed in 877 Korean asthmatics. This study found that several new genes might be associated with total IgE in asthmatics, such as CRIM1 (rs848512, P = 1.18×10⁻⁶; rs711254, P = 6.73×10⁻⁶), ZNF71 (rs10404342, P = 7.60×10⁻⁶), TLN1 (rs4879926, P = 7.74×10⁻⁶), and SYNPO2 (rs1472066, P = 8.36×10⁻⁶; rs1038770, P = 8.66×10⁻⁶). Regarding the association of specific IgE to house dust mites, it was observed that intergenic SNPs nearby to OPRK1 and LOC730217 might be associated with Dermatophagoides pteronyssinus (D.p.) and Dermatophagoides farinae (D.f.) in asthmatics, respectively. In further pathway analysis, the phosphatidylinositol signaling system and adherens junction pathways were estimated to play a role in the regulation of total IgE levels in asthma. Although functional evaluations and replications of these results in other populations are needed, this GWAS of serum IgE in asthmatics could facilitate improved understanding of the role of the newly identified genetic variants in asthma and its related phenotypes.     

Genome-Wide Association Studies Identify Two Novel BMP15 Mutations Responsible for an Atypical Hyperprolificacy Phenotype in Sheep

Some sheep breeds are naturally prolific, and they are very informative for the studies of reproductive genetics and physiology. Major genes increasing litter size (LS) and ovulation rate (OR) were suspected in the French Grivette and the Polish Olkuska sheep populations, respectively. To identify genetic variants responsible for the highly prolific phenotype in these two breeds, genome-wide association studies (GWAS) followed by complementary genetic and functional analyses were performed. Highly prolific ewes (cases) and normal prolific ewes (controls) from each breed were genotyped using the Illumina OvineSNP50 Genotyping Beadchip. In both populations, an X chromosome region, close to the BMP15 gene, harbored clusters of markers with suggestive evidence of association at significance levels between 1E⁻⁰⁵ and 1E⁻⁰⁷. The BMP15 candidate gene was then sequenced, and two novel non-conservative mutations called FecX^Gr and FecX^O were identified in the Grivette and Olkuska breeds, respectively. The two mutations were associated with the highly prolific phenotype (p_FecX^Gr = 5.98E⁻⁰⁶ and p_FecX^O = 2.55E⁻⁰⁸). Homozygous ewes for the mutated allele showed a significantly increased prolificacy (FecX^Gr/FecX^Gr, LS = 2.50±0.65 versus FecX⁺/FecX^Gr, LS = 1.93±0.42, p<1E⁻⁰³ and FecX^O/FecX^O, OR = 3.28±0.85 versus FecX⁺/FecX^O, OR = 2.02±0.47, p<1E⁻⁰³). Both mutations are located in very well conserved motifs of the protein and altered the BMP15 signaling activity in vitro using a BMP-responsive luciferase test in COV434 granulosa cells. Thus, we have identified two novel mutations in the BMP15 gene associated with increased LS and OR. Notably, homozygous FecX^Gr/FecX^Gr Grivette and homozygous FecX^O/FecX^O Olkuska ewes are hyperprolific in striking contrast with the sterility exhibited by all other known homozygous BMP15 mutations. Our results bring new insights into the key role played by the BMP15 protein in ovarian function and could contribute to a better understanding of the pathogenesis of women′s fertility disorders.     

Determination of Critical Nitrogen Dilution Curve Based on Stem Dry Matter in Rice

Plant analysis is a very promising diagnostic tool for assessment of crop nitrogen (N) requirements in perspectives of cost effective and environment friendly agriculture. Diagnosing N nutritional status of rice crop through plant analysis will give insights into optimizing N requirements of future crops. The present study was aimed to develop a new methodology for determining the critical nitrogen (N_c) dilution curve based on stem dry matter (S_DM) and to assess its suitability to estimate the level of N nutrition for rice (Oryza sativa L.) in east China. Three field experiments with varied N rates (0–360 kg N ha⁻¹) using three Japonica rice hybrids, Lingxiangyou-18, Wuxiangjing-14 and Wuyunjing were conducted in Jiangsu province of east China. S_DM and stem N concentration (SNC) were determined during vegetative stage for growth analysis. A N_c dilution curve based on S_DM was described by the equation (N_c = 2.17W^−0.27with W being S_DM in t ha⁻¹), when S_DM ranged from 0.88 to 7.94 t ha⁻¹. However, for S_DM < 0.88 t ha⁻¹, the constant critical value N_c = 1.76% S_DM was applied. The curve was dually validated for N-limiting and non-N-limiting growth conditions. The N nutrition index (NNI) and accumulated N deficit (N_and) of stem ranged from 0.57 to 1.06 and 51.1 to −7.07 kg N ha⁻¹, respectively, during key growth stages under varied N rates in 2010 and 2011. The values of ΔN derived from either NNI or N_and could be used as references for N dressing management during rice growth. Our results demonstrated that the present curve well differentiated the conditions of limiting and non-limiting N nutrition in rice crop. The S_DM based N_c dilution curve can be adopted as an alternate and novel approach for evaluating plant N status to support N fertilization decision during the vegetative growth of Japonica rice in east China.