Natural killer cell IFN-γ-activity is associated with Plasmodium falciparum infection during pregnancy

The ability of natural killer (NK) cells to produce gamma interferon (IFN-γ) after ex vivo stimulation with crude schizont lysate of Plasmodium falciparum was studied in uninfected and P. falciparum-infected pregnant Gabonese women segregated according to the gravidity at the time of delivery. This activity was measured in purified NK cells as well as in whole blood from the periphery and cord. Crude schizont lysate-stimulated NK cells from primiparous women produced significantly more IFN-γ than those from multiparous women (P < 0.001). Women with malaria infection produced more IFN-γ than negative women in peripheral blood (P < 0.001) indicating that immunological determinants regulating the susceptibility to malaria in pregnant women are parasite-specific. These findings reveal that NK cells are major source of IFN-γ when exposed to P. falciparum antigens in vitro in absence of any other co-stimulant.     

Characteristics of the spermathecal contents of old and young honeybee queens

Sperm are often stored, for a long time after mating, in females of various animal species. In case of the queen honeybee (Apis mellifera), sperm remain fertile for several years in the spermatheca. Little information is available regarding the effect of long-term storage of sperm on its fertility. To evaluate this, enzymes and/or sperm have been analysed from the spermatheca of 75 queens of various ages (0 year Y0, n = 14; one year Y1, n = 14; two years Y2, n = 7; virgin queen VQ, n = 40) and semen samples have been taken from 46 drones. The sperm from the spermatheca of older queens move more slowly (F = 11.45, P < 0.0001) and show different movement patterns (Chi² = 90.0, P < 0.0001) from those of the other groups. The spermatheca content of differently aged mated queens differ significantly with respect to the activities of lactate dehydrogenase (F = 3.37, P < 0.05), citrate synthase (F = 6.24, P < 0.005) and arginine kinase (F = 9.44, P < 0.0006). Glyceraldehyde 3-phosphate dehydrogenase (F = 0.10, P = 0.91) does not differ significantly. The results suggest considerable changes in the energy metabolic profile of the spermatheca tissue, of the sperm or of both during sperm storage.     

Photonic characteristics and ex vivo imaging of Escherichia coli-Xen14 within the bovine reproductive tract

The objectives of this study were to (1) characterize the photonic properties of Escherichia coli-Xen14 and (2) conduct photonic imaging of E. coli-Xen14 within bovine reproductive tract segments (RTS) ex vivo (Bos indicus). E. coli-Xen14 was grown for 24 h in Luria Bertani medium (LB), with or without kanamycin (KAN). Every 24 h, for an 8-d interval, inoculums were imaged and photonic emissions (PE) collected. Inoculums were subcultured and plated daily to determine the colony forming units (CFU) and ratio of photon emitters to nonemitters. In the second objective, abattoir-derived bovine reproductive tracts (n = 9) were separated into posterior and anterior vagina, cervix, uterine body, and uterine horns. Two concentrations (3.2 × 10⁸ and 3.2 × 10⁶ CFU/200 μL for relative [High] and [Low], respectively) of E. coli-Xen14 were placed in translucent tubes for detection of PE through RTS. The CFU did not differ (P = 0.31) over time with or without KAN presence; they remained stable with 99.93% and 99.98% photon emitters, respectively. However, PE were lower (P < 0.0001) in cultures containing KAN than in those containing no KAN (629.8 ± 117.7 vs. 3012.0 ± 423.5 relative lights units per second [RLU/sec], respectively). On average, the percentage of PE between RTS, for both concentrations, was higher (P < 0.05) in the uterine body. In summary, E. coli-Xen14 remained stable with respect to the proportions of photon emitters with or without KAN (used to selectively culture E. coli-Xen14). However, KAN presence suppressed photonic activity. The ability to detect PE through various segments of the reproductive tract demonstrated the feasibility of monitoring the presence of E. coli-Xen14 in the bovine reproductive tract ex vivo.     

A novel in vivo regulatory role of P-glycoprotein in alloimmunity

P-glycoprotein (P-gp) is required for adaptive immunity through defined functions in T cell activation and antigen presenting cell (APC) maturation. The potential role of P-gp as an in vivo regulator of alloimmunity is currently unknown. Here we show that P-gp blockade prolongs graft survival in a murine heterotopic cardiac allotransplantation model through in vivo inhibition of the T helper 1 (Th1) cytokine IFN-γ and the Th2 product IL-4, and via downregulation of the APC-expressed positive costimulatory molecule CD80. In vitro, the P-gp antagonist PSC833, a non-calcineurin-inhibitory cyclosporine A analogue, specifically inhibited cellular efflux of the P-gp substrate rhodamine-123 in wild-type CD3⁺ T cells and MHC class II⁺ APCs but not their P-gp knockout counterparts that lacked rhodamine-123 efflux capacity. Additionally, P-gp blockade significantly inhibited murine alloimmune T cell activation in a dose-dependent fashion. In vivo, P-gp blockade significantly prolonged graft survival in Balb/c recipients of C57BL/6 cardiac allografts from 8.5 ± 0.5 to 11.7 ± 0.5 days (P < 0.01), similar in magnitude to the effects of monotherapy with cyclosporine A. Moreover, P-gp blockade, compared to controls, attenuated intragraft expression of CD3 and CD80, but not CD86, and inhibited IFN-γ and IL-4 production (P < 0.05). In the setting of systemic CD86 inhibition, P-gp blockade suppressed IFN-γ and IL-4 production significantly further (to 98% and 89% inhibition, respectively) compared to either P-gp or anti-CD86 blockade alone, and markedly prolonged allograft survival compared to anti-CD86 blockade alone (40.5 ± 4.6 versus 22.5 ± 2.6 days, respectively, P < 0.01). Our findings define a novel in vivo regulatory role of P-gp in alloimmunity and identify P-gp as a potential therapeutic target in allotransplantation.     

Regulation of carboxylesterase 1 (CES1) in human adipose tissue

Carboxylesterase 1 (CES1) has recently been suggested to play a role in lipolysis. Our aim was to study the regulation of CES1 expression in human adipose tissue. In the SOS Sib Pair Study, CES1 expression was higher in obese compared with lean sisters (n = 78 pairs, P = 8.7 × 10⁻¹⁸) and brothers (n = 12 pairs, P = 0.048). CES1 expression was higher in subcutaneous compared with omental adipose tissue in lean (P = 0.027) and obese subjects (P = 0.00036), and reduced during diet-induced weight loss (n = 24, weeks 8, 16, and 18 compared to baseline, P < 0.0001 for all time points). CES1 expression was higher in isolated adipocytes compared with intact adipose tissue (P = 0.0018) and higher in large compared with small adipocytes (P = 4.1 × 10⁻⁶). Basal and stimulated lipolysis was not different in individuals with high, intermediate, and low expression of CES1. Thus, CES1 expression was linked to body fat and adipocyte fat content but not to lipolytic activity.     

An experimental facility for free air humidity manipulation (FAHM) can alter water flux through deciduous tree canopy

A facility for free air humidity manipulation (FAHM) was established to investigate the effect of increased air humidity on trees’ performance and their canopy functioning with respect to rising air humidity predicted for Northern Europe. The FAHM system enables air relative humidity (RH) to be increased up to 18 units (%) over the ambient level during mist fumigation, depending on the wind speed inside the experimental stand. Water was dispersed inside 14 × 14 m experimental plots in the form of mist with an average particle size of 50 μm from June to August in 2008, and from May to September in 2009. The average increase in RH was 7 units (%) over the whole period of humidification in 2008 (P < 0.05). The average diurnal stem sap flux density per unit projected leaf area (F) in silver birch (Betula pendula Roth.) trees was 24.8% (P < 0.05) and 27.2% (P < 0.01) higher in control (C) plots compared to humidification (H) plots during misting in 2008 and 2009, respectively. However, the difference between C and H plots was statistically insignificant (P > 0.05) in silver birch on the days without misting. In hybrid aspen (Populus tremula L. × P. tremuloides Michx.) the average difference in F between C and H plots was 61.1% (P < 0.001) during mist fumigation in the summer of 2009. Nevertheless, the difference was considerable (38.8%; P < 0.001) also on the days without misting, reflecting the impact of plant inner factors on F as a result of long-term acclimation to fumigation. The leaves of silver birch in a humidified plot demonstrated up to 2.4 °C lower (P < 0.05) leaf temperature (T_L) compared to the control plot in 2009. The decline in T_L decreased the humidity gradient between leaf and air by about 1/3, whereas 2/3 of the effect was caused directly by changes in air humidity in the leaf boundary layer. Our preliminary data suggest that the FAHM experimental facility enables water fluxes through a deciduous tree canopy to be reduced and this effect is attributable both to the increased air humidity and decreased leaf temperature. Changes in these two basic factors may create considerable differences in the physiology, anatomy and nutrition of a whole tree, also affecting forest functioning in the light of global climate change.     

Cox-2 gene variants in migraine

Purpose

Migraine is a multifactorial and complex disorder, and any clear diagnostic marker to assess the status of the migraineurs has not been established, yet. Nonsteroidal anti-inflammatory drugs reduce production of prostanoids including PGE2 by inhibiting COX-1 and/or COX-2, and thereby suppress inflammatory pain in patients suffering from rheumatoid arthritis, osteoarthritis, and migraine. Thus, COX-2 regulation is important in the pathogenesis and treatment of migraine. We prospectively investigated COX-2-765G → C and COX-2-1195A → G gene polymorphisms which may account for an increased risk of migraine.

Methods

The present analyses are based on 144 case subjects with migraine disease and 123 non-case subjects. Genotyping of COX-2 gene polymorphisms (COX-2-765G → C, COX-2-1195A → G) was detected by PCR-RFLP.

Results

We, for the first time, demonstrated positive association of COX-2 gene variants with an increased risk for development of migraine. Carriers of COX-2-765 C + genotype in controls were higher than in the patients (57.7% and 36.1% respectively; P < 0.0001) and the frequencies of G + genotype in patients were higher than in the controls (97.9% and 88.6% respectively; P: 0.002). In addition, frequencies of COX-2-765 GG and GC genotypes in patients were higher than in the controls (P < 0.0001, P < 0.0001 respectively). It seems that COX-2-765 G + genotype had increased and COX-2-765 C + genotype had decreased risk for migraine. In COX-2-1195 polymorphism only AG genotype was statistically significantly different in patients than in the controls (P < 0.05).

Conclusions

Our findings have suggested that COX-2-765 G + genotype could facilitate the development of migraine disease.     

Cytotoxicity from sulfide exposure in a sulfide-tolerant marine invertebrate

It is unknown whether sulfide-tolerant marine invertebrates suffer cytotoxicity from sulfide exposure in vivo at environmentally-relevant concentrations. We tested this with the mudflat polychaete Glycera dibranchiata. Exposure of live animals to sulfide up to 2.4 mmol l^− 1 for 24 h did not affect animal survival, and animal condition (gross appearance and activity) was unaffected at sulfide concentrations up to 0.25 mmol l^− 1. However, animal condition was decreased at higher sulfide concentrations (P < 0.0001, n = 33). Coelomic fluid obtained from the animals showed decreased erythrocyte count (P = 0.0035, n = 14), indicating cell loss, and increased propidium iodide and Hoechst 33324 staining (P < 0.0001 and P = 0.0003, respectively, n = 14), indicating loss of plasma membrane integrity, even at sulfide concentrations that produced no change in animal condition. When G. dibranchiata were allowed a 72 h recovery period following 24 h sulfide exposure, there was no overall improvement in condition (P ≥ 0.12, n = 7-8), and worms that had been exposed to 1 mmol l^−  1 sulfide still had an erythrocyte count that was less than half that of control animals (P = 0.028, n = 7). The inability to completely recover the cell count was at least partially due to impaired production of new erythrocytes, since analysis of BrdU incorporation indicated that erythrocyte proliferation was reduced from 2% per day in control animals to 0.12% per day in animals exposed to 1 mmol l^− 1 sulfide (P = 0.010, n = 21). Together, these findings indicate that at least some sulfide-tolerant marine invertebrates experience significant cellular injury and impaired tissue proliferation when exposed to environmentally relevant sulfide concentrations, even when the appearance and behavior of the animal appear unaffected.     

Citric acid improves growth performance and phosphorus digestibility in Beluga (Huso huso) fed diets where soybean meal partly replaced fish meal

A 2 × 3 factorial experiment was conducted to evaluate the effect of partial substitution of dietary fish meal with soybean meal and citric acid (CA) supplementation on growth, food utilization, muscle composition and nutrient digestibility of Beluga, Huso huso. Three isonitrogenic and isoenergetic diets, as SBM₁ (soybean meal protein (SBP):fishmeal protein (FP) = 1:3), SBM₂ (SBP:FP = 2:3) and SBM₃ (SBP:FP = 1:1) containing two levels of CA (0 and 30 g kg⁻¹) were fed to triplicate groups of fish for 8 weeks. The results revealed that adding CA increased (P<0.05) the weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) and decreased (P<0.05) food conversion rate (FCR) whereas partial substitution of fishmeal with soybean meal decreased (P<0.05) growth performance. Partial replacement of fishmeal with SBM (P=0.021) as well as CA supplementation (P=0.019) and their interaction (P<0.001) affected hepatosomatic index (HSI). No differences (P>0.05) were observed for viscerosomatic index (VSI) among treatments (P>0.05). No differences (P>0.05) were detected in moisture, protein of muscle sample among treatments, but lipid content was reduced (P<0.05) while ash content increased (P=0.035) by CA. Soybean meal decreased (P<0.05) nutrient digestibility, whereas CA improved apparent protein and phosphorus (P) digestibility (P=0.011 and P<0.001 respectively). No interaction (P<0.05) between levels of SBM and CA was found on these parameters. Results of the present study indicate that Beluga has a limited ability to utilize SBM as a protein source in practical diets whereas CA can improve growth and nutrient utilization in Beluga.     

The modification of glucose levels and N source in the Hungate's medium to stimulate the production of fibrolytic enzymes of Anaeromyces sp. YQ3 grown on corn stalks

Hungate's method is a well-accepted protocol for the isolation or incubation of anaerobes with a roll tube technique. The aim of this study was to stimulate fungal enzyme production by optimizing the components of Hungate's medium for the growth of a rumen fungus Anaeromyces sp. YQ3. The organism was grown on corn stalks and incubated for 10 days in defined media with two glucose levels (G⁺, glucose in the Hungate's medium as a glucose control; G^?, glucose removed in a modified Hungate's medium) and four N sources (N1: yeast extract + tryptone + (NH₄)₂SO₄ in Hungate's medium (control); N2: yeast extract + (NH₄)₂SO₄; N3: tryptone + (NH₄)₂SO₄; and N4: tryptone + yeast extract). In the G^? media, the recovered activities of feruloyl esterase (FAE) (P<0.0001), acetyl esterase (AE) (P=0.0065) and xylanase (P<0.0001) were decreased, while the G⁺ media with N1 nitrogen stimulated the production of FAE and xylanase (P<0.0001). The G^? medium with N2 nitrogen increased the recovered activities of carboxymethyl cellulase (P=0.0001) and avicelase (P<0.0001), while the N3 and N4 media increased the recovered activity of AE (P=0.0015). The N4 medium was comparable to the N1 medium in stimulating the amount of recovered xylanase activity. The activities of FAE (P<0.0001), AE (P<0.0001), and xylanase (P<0.0001) showed a time-dependent increase and reached their peaks at day 10, while the avicelase activity peaked at day 8 (P=0.0071). The esterase activities (FAE and AE) were positively correlated with the enzyme activities of xylanase and carboxymethyl cellulase (r > 0.48, P<0.05). After a 10-day incubation, the glucose in the Hungate's media contributed to an increase in organic matter disappearance (P<0.0001) and volatile fatty acid (VFA) concentration (P<0.0001), except for molar acetate proportions. The N4 treatment increased organic matter disappearance and total VFA concentration (P=0.0002). The change in N source did not alter molar proportions of acetate, propionate and valerate, while the N2 treatment increased molar butyrate proportion (P<0.0035), and both N2 and N3 increased the molar proportion of branched chain VFAs (P<0.0041). In summary, the glucose in the Hungate's medium is beneficial for stimulating the production of esterases and xylanase, thereby promoting fungal growth. Amending the N source in Hungate's medium brings about different yields of rumen fungal esterases and polysaccharide hydrolases that have important nutritional impacts on fibre degradation in ruminant animals.     

The relationship between five widely-evaluated variants in CDKN2A/B and CDKAL1 genes and the risk of type 2 diabetes: A meta-analysis

The genes encoding two cyclin-dependent kinases-inhibitor-2A/B (CDKN2A/B) and 5 regulatory subunit-associated protein-like 1 (CDKAL1) have been investigated extensively in associations with type 2 diabetes; the results, however, are often irreproducible. We therefore sought to evaluate these associations by performing a meta-analysis on five widely-evaluated variants from the two genes. There were 38 studies (patients/controls: 51,940/52,234) for rs10811661, 16 studies (20,029/24,419) for rs564398 in CDKN2A/B gene, and 27 studies (28,383/47,635) for rs7756992, 26 studies (28,816/31,713) for rs7754840, 21 studies (29,260/38,400) for rs10946398 in CDKAL1 gene. Overall risk estimates for type 2 diabetes conferred by rs10811661-T, rs564398-A, rs7754840-C, rs7756992-G, and rs10946398-C alleles were 1.17 (95% CI: 1.10–1.23; P < 0.0005; I² = 83.9%), 1.1 (95% CI: 1.0–1.21; P = 0.051; I² = 88.3%), 1.24 (95% CI: 1.18–1.3; P < 0.0005; I² = 74.3%), 1.2 (95% CI: 1.11–1.3; P < 0.0005; I² = 92.0%), and 1.19 (95% CI: 1.1–1.29; P < 0.0005; I² = 90.8%), respectively. There was evident publication bias for rs564398 and rs7754840. Subgroup analyses by ethnicity showed remarkable divergences in risk estimate for rs564398 between Asians (odds ratio [OR] = 1.01; 95% CI: 0.86–1.19; P = 0.868) and Caucasians (OR = 1.19; 95% CI: 1.03–1.35; P = 0.012) (P < 0.05). For all variants examined, the results of studies in retrospective design or with population-based controls were comparative with that of overall studies. In meta-regression analyses, age was found to exert a significant influence on the association between rs10811661 and type 2 diabetes (P = 0.003), as well as between rs7754840 and gender (P = 0.034). Taken together, our findings provide evidence for a significant contribution of CDKN2A/B gene rs10811661 and CDKAL1 gene rs7756992 and rs10946398 to type 2 diabetes.     

滞育诱导温周期对桃小食心虫滞育幼虫生理指标的影响

【目的】为了明确秋季温度升高对桃小食心虫Carposina sasakii Matsumura 耐寒性的影响,针对桃小食心虫滞育过程的生理指标进行了一系列研究。【方法】在GXZ-0450型智能光照培养箱中,温度为26~18, 26~20, 26~22, 28~18, 28~20, 28~22, 30~18, 30~20和30~22℃ 9组变温下（高温10 h,低温14 h,相对湿度70%~80%,光周期为12L∶12D）分别诱导桃小食心虫滞育,再对滞育幼虫过冷却点（supercooling point, SCP）、结冰点（freezing point, FP）以及体内水、脂肪和小分子糖醇类物质含量进行测定。【结果】在适宜滞育的光照下变温不会影响桃小食心虫的滞育率,所有脱果的老熟幼虫全部进入滞育,但其生理指标却有很大差异,其中SCP和FP在较高日高温下较高（SCP：F_135,2=23.83,P<0.001;FP：F_135,2=47.64,P<0.0001）,而在较高夜低温下反而较低（SCP：F_135,2=52.88,P<0.0001;FP：F_135,2=33.34,P<0.0001）。含水量随夜低温和日高温升高显著升高（夜低温：F_135,2=13.47,P<0.0001;日高温：F_135,2=10.39,P<0.0001）。脂肪含量随夜低温升高下降（F_135,2=40.91, P<0.0001）,随日高温升高而上升（F_135,2=161.18,P<0.0001）。小分子糖醇类物质含量随温度的变化各不相同,但差异均显著（P<0.0001）：其中海藻糖、葡萄糖和赤藓糖醇的含量随夜低温升高而下降,但在日高温30℃时海藻糖和赤藓糖醇的含量却随夜温的升高而升高,葡萄糖的含量随夜低温的变化无差异;甘油随日高温的升高而显著升高,随夜低温的升高而显著下降;山梨醇和肌醇的含量随夜低温和日高温的变化差异很大。【结论】桃小食心虫不同温度下诱导滞育的幼虫SCP、FP、水、脂肪和小分子糖醇类含量存在明显差异。滞育诱导期温度对桃小食心虫幼虫的滞育生理有重要的作用,其滞育幼虫起始生理差别可能会影响其越冬。     

The effects of saliva collection,handling and storage on salivary testosterone measurement

Several endocrine parameters commonly measured in plasma, such as steroid hormones, can be measured in the oral fluid. However, there are several technical aspects of saliva sampling and processing that can potentially bias the validity of salivary testosterone measurement. The aim of this study was to evaluate the effects caused by repeated sampling; 5 min centrifugation (at 2000, 6000 or 10,000g); the stimulation of saliva flow by a cotton swab soaked in 2% citric acid touching the tongue; different storage times and conditions as well as the impact of blood contamination on salivary testosterone concentration measured using a commercially available ELISA kit. Fresh, unprocessed, unstimulated saliva samples served as a control. Salivary testosterone concentrations were influenced neither by repeated sampling nor by stimulation of salivary flow. Testosterone levels determined in samples stored in various laboratory conditions for time periods up to 1 month did not differ in comparison with controls. For both genders, salivary testosterone levels were substantially reduced after centrifugation (men F = 29.1; women F = 56.17, p < 0.0001). Blood contamination decreased salivary testosterone levels in a dose-dependent manner (men F = 6.54, p < 0.01, F = 5.01, p < 0.05). Salivary testosterone can be considered A robust and stable marker. However, saliva processing and blood leakage can introduce bias into measurements of salivary testoterone using ELISA. Our observations should be considered in studies focusing on salivary testosterone.     

Effects of NaOH treatment on condensed tannin contents and gas production kinetics of tree leaves

Effects of NaOH treatment on the crude protein (CP), condensed tannin (CT) and in vitro gas production kinetics of leaves of Arbutus andrachne, Glycyrrhiza glabra L. and wheat straw were determined. Wheat straw, which is tannin-free, was used as the standard. The NaOH treatment was completed by pulverization of samples with 0, 20, 40, 60 and 80 g/L of NaOH solution in the proportion of 1 L of solution to 1 kg of sample. Gas production was determined at 3, 6, 12, 24, 48, 72 and 96 h of incubation. NaOH treatment linearly decreased (P<0.001) the CT contents of leaves of Arbutus andrachne and Glycyrrhiza glabra L. whereas NaOH treatment had no effect on the CP contents of Arbutus andrachne, Glycyrrhiza glabra L. and wheat straw. The 80 g/L NaOH treatment reduced the CT content of leaves of Arbutus andrachne and Glycyrrhiza glabra L. by 59.6% and 86.7%, respectively. NaOH treatment linearly decreased (P<0.01) gas production rate of Arbutus andrachne although it linearly increased (P<0.01) gas production rate of wheat straw. In contrast, NaOH treatment had no effect on gas production rate of leaves of Glycyrrhiza glabra L. NaOH treatment linearly decreased (P<0.001) potential gas production of leaves of Arbutus andrachne and Glycyrrhiza glabra L. whereas NaOH treatment linearly increased (P<0.001) potential gas production of wheat straw. Treatment with NaOH can be used to improve the nutritive value of tannin-free forages such as straw, but not for tannin-containing leaves.     

In vitro ruminal methanogenesis of a hay-rich substrate in response to different combination supplements of nitrocompounds; pyromellitic diimide and 2-bromoethanesulphonate

Sixteen combinations of 5 treatments at 4 levels were designed in a L₁₆(4⁵) orthogonal experimental design to evaluate associative effects of five methanogenesis inhibitors at four dose levels: nitroethane (NE, 0 mM, 5 mM, 10 mM and 15 mM), 2-nitroethanol (NEOH, 0 mM, 5 mM, 10 mM and 15 mM), 2-nitro-1-propanol (NPOH, 0 mM, 5 mM, 10 mM and 15 mM), pyromellitic diimide (PMDI, 0 mM, 0.02 mM, 0.05 mM and 0.07 mM) and 2-bromoethanesulphonate (BES, 0 mM, 0.01 mM, 0.03 mM and 0.05 mM) on in vitro ruminal methane production of the mixed substrate (Chinese wildrye hay:maize meal = 4:1) using a cumulative gas production technique. After 48 h incubation, in vitro dry matter disappearance (IVDMD), total gas production (GP₄₈, ml/g DM) and total volatile fatty acids (VFA) production in various combinations of these inhibitors were decreased by 10.6-56.0, 26.5-44.5 and 20.3-47.6%, respectively (P<0.05). The molar proportion of acetate in the inhibitor combination groups was decreased by 6.6-12.5% while those of propionate and butyrate were increased by 7.0-19.2 and 21.9-56.5% (P<0.01), respectively. Methane proportion (MP) in total gas production was reduced by 79.4-98.5% (P<0.01), and the highest inhibition occurred in the combination of 10 mM NE, 10 mM NPOH, 0.07 mM PMDI and 0.01 mM BES in cultures. The partial correlation coefficients between NE, NEOH, NPOH, PMDI or BES and CH₄ proportion were −0.465 (P<0.01), −0.417 (P<0.01), −0.355 (P<0.05), −0.408 (P<0.01) and −0.345 (P<0.05), respectively, indicating that NE was the most potent inhibitor, followed by NEOH and PMDI, and finally NPOH and BES. In general, VFA production in the inhibitor combinations was substantially shifted to produce much more butyrate and propionate and less acetate. The combination of 15 mM NE, 10 mM NEOH, 5 mM NPOH, 0.07 mM PMDI and 0.01 mM BES in cultures, leading to >95% methane inhibition, may be the optimal application of these inhibitors with less depression of total VFA production. Further feeding trials to validate these combinations is still required on rumen function, methane production, growth performance and milk production.     

Application of computed tomography for the evaluation of obstetrically relevant pelvic parameters in German Holstein-Friesian cows

The aim of this study was to use computed tomography for the evaluation of the suitability of external pelvimetry to determine obstetrically relevant parameters. External pelvimetric measurements obtained in vivo using calipers and internal pelvimetric measurements obtained in vitro using computed tomography were taken in 30 German Holstein-Friesian cows (Bos taurus). All measurements were highly reproducible with intraclass correlation coefficients ≥98%. Hip width was the external variable with the highest correlation to internal variables, particularly pelvic inlet area and circumference, pelvic volume, medial horizontal diameter of the pelvic inlet, and the narrowest horizontal diameter of the midpelvis (r² > 0.60, P < 0.0001). The pelvic inlet area and circumference, the pelvic volume, and the diagonal diameter of the pelvis were sufficiently predicted with the aid of external pelvic measurements and age (r²≥ 0.80, P < 0.0001). The results of this study show that external pelvimetry yields reliable information about the size of the pelvis when the age of the cow is considered.     

New cerebellar phenotypes in YAC transgenic mouse in vivo library of human Down syndrome critical region-1

Down syndrome (DS) is the most frequent genetic cause of mental retardation (MR) associated with neurological alterations. To allow a genetic dissection of DS phenotype, we studied eight transgenic mouse lines carrying YACs containing human DNA fragments covering DS critical region (DCR-1), as an in vivo library. Herein, we found an increased brain size in the 152F7-mice containing DYRK1A gene. We also identified a new cerebellar alteration in two independent lines carrying 230E8-YAC. These mice showed significant elongation of the cerebellar antero-posterior axis (p < 0.001), determined by increased length of rostral folia of the vermis (lobule II-V, p < 0.0001; lobule VI, p < 0.001). In addition, we identified a major neurological defect in culmen and declivus lobules in the 230E8-mice. We analyzed P30, P12, and P9 stages and detected high significant increased lengths of anterior lobules (II-VI) of 230E8-mice at P30 and P12 (lobule II-V, p < 0.0001; lobule VI, p < 0.05), but not at P9, indicating that this new phenotype appears between P9 and P12. Interestingly, 230E8-mice also present increased cortical cell density and mild learning defects. 230E8-YAC contains seven genes, some of which could be potentially responsible for this phenotype. Between them, we proposed DOPEY2 as potential candidate gene for these cerebellar alterations considering its high expression in the brain and that its homologous genes in yeast, Caenorhabditis elegans and Drosophila are involved in morphogenesis, suggesting a conserved role of DOPEY2 as a patterning gene.     

Egg production and early development of Thysanoessa inermis and Euphausia pacifica (Crustacea: Euphausiacea) in the northern Gulf of Alaska

Early life history patterns were studied in the dominant euphausiids from the northern Gulf of Alaska (GOA) in 2001-2004. Gravid females of Thysanoessa inermis were observed in April and May. Brood size varied from 10 to 1021 eggs with an average of 138 ± 19 (95% CI) eggs female^− 1. Most gravid females started to release eggs within the first 2 days of incubation. The average number of eggs released per female was similar in incubation Day 1 and 2, but significantly smaller on Day 3 and 4. About 25% of the females were continuously releasing eggs over 3 days rather than producing a single distinctive brood. In contrast, gravid females of Euphausia pacifica were observed from early July through October. Most gravid females released eggs on the first day of observation, while only 2% of females produced eggs repeatedly. Brood size varied from 20 to 246 eggs with an average of 102 ± 12 (95% CI) eggs female^− 1. The relationship between E. pacifica brood size and ambient chlorophyll-a concentration was sigmoidal (r² = 0.73), with food saturated brood size of 144 ± 14(SE, P < 0.001) eggs, and half-saturation occurring at 0.46 ± 0.02(SE, P < 0.001) mg chlorophyll-a m^− 3. The average interbrood interval of E. pacifica reared at 12 °C and satiated food conditions in the laboratory was ∼ 8 days, suggesting their potential individual fecundity in the GOA was 1148-1530 eggs per spawning season. Hatching and early development (from egg to furcilia stage) was studied under 5 °C, 8 °C and 12 °C. Hatching was nearly synchronous and lasted 3-6 h, depending on incubation temperature. Development times from egg to the first furcilia stage ranged between 20 and 33 days for T. inermis, and 15 and 45 days for E. pacifica at 12 °C and 5 °C, respectively.