Association of ovarian response with picoAMH in women undergoing controlled ovarian hyperstimulation.

OBJECTIVE: Our study aims to evaluate the diagnostic performance of a high-sensitivity picoAnti-Müllerian Hormone (picoAMH) for predicting ovarian response in women undergoing controlled ovarian hyperstimulation and occurrence of ovarian hyperstimulation syndrome. METHODS: Retrospective cohort study at a single academic fertility center including all patients with picoAMH ELISA who underwent controlled ovarian hyperstimulation. The primary outcome was the number of oocytes retrieved, and secondary outcomes included cycle cancellation and ovarian hyperstimulation syndrome. Patients were grouped into poor, normal, and hyper-responders based on number of oocytes retrieved. RESULTS: The mean AMH and antral follicle count (AFC) were significantly different between normal response vs. hyper response group (p < 0.0001). Only serum AMH and not AFC was significantly increased in patients diagnosed with ovarian hyperstimulation syndrome (OHSS). For prediction of OHSS, receiver operating characteristic (ROC) analysis revealed that AMH (area under the ROC curve [AUC] = 0.85) was significantly better than the AFC (AUC = 0.64). The serum AMH cut-off at sensitivity of 80% for predicting OHSS among hyper responders from ROC curve was 3.67 ng/ml. Serum AMH measured by picoAMH ELISA showed superior correlation to number of oocytes retrieved when compared to AFC in the age group over 40 years old (r2 = 0.74 and r2 = 0.4, respectively) CONCLUSION: This study shows great utility of picoAMH ELISA for predicting ovarian response to controlled ovarian hyperstimulation (COH). Diagnostic performance of picoAMH for prediction of OHSS is superior to the AFC in our cohort.

Comparative analysis of the ileal bacterial composition of post-weaned pigs fed different high-quality protein sources.

To further understand the contribution of feedstuff ingredients to gut health in swine, gut histology and intestinal bacterial profiles associated with the use of two high-quality protein sources, microbially enhanced soybean meal (MSBM) and Menhaden fishmeal (FM) were assessed. Weaned pigs were fed one of three experimental diets: (1) basic diet containing corn and soybean meal (Negative Control (NEG)), (2) basic diet + fishmeal (FM; Positive Control (POS)) and (3) basic diet + MSBM (MSBM). Phase I POS and MSBM diets (d 0 to d 7 post-wean) included FM or MSBM at 7.5%, while Phase II POS and MSBM diets (d 8 to d 21) included FM or MSBM at 5.0%. Gastrointestinal tissue and ileal digesta were collected from euthanised pigs at d 21 (eight pigs/diet) to assess gut histology and intestinal bacterial profiles, respectively. Data were analysed using Proc Mixed in SAS, with pig as the experimental unit and pig (treatment) as the random effect. Histological and immunohistochemical analyses of stomach and small intestinal tissue using haematoxylin-eosin, Periodic Acid Schiff/Alcian blue and inflammatory cell staining did not reveal detectable differences in host response to dietary treatment. Ileal bacterial composition profiles were obtained from next-generation sequencing of PCR generated amplicons targeting the V1 to V3 regions of the 16S rRNA gene. Lactobacillus-affiliated sequences were found to be the most highly represented across treatments, with an average relative abundance of 64.0%, 59.9% and 41.80% in samples from pigs fed the NEG, POS and MSBM diets, respectively. Accordingly, the three most abundant Operational Taxonomic Units (OTUs) were affiliated to Lactobacillus, showing a distinct abundance pattern relative to dietary treatment. One OTU (SD_Ssd_00001), most closely related to Lactobacillus amylovorus, was found to be more abundant in NEG and POS samples compared to MSBM (23.5% and 35.0% v. 9.2%). Another OTU (SD_Ssd_00002), closely related to Lactobacillus johnsonii, was more highly represented in POS and MSBM samples compared to NEG (14.0% and 15.8% v. 0.1%). Finally, OTU Sd_Ssd-00011, highest sequence identity to Lactobacillus delbrueckii, was found in highest abundance in ileal samples from MSBM-fed pigs (1.9% and 3.3% v. 11.3, in POS, NEG and MSBM, respectively). There was no effect of protein source on bacterial taxa to the genus level or diversity based on principal component analysis. Dietary protein source may provide opportunity to enhance presence of specific members of Lactobacillus genus that are associated with immune-modulating properties without altering overall intestinal bacterial diversity.

Congenital Uterine Malformation by Experts (CUME): better criteria for distinguishing between normal/arcuate and septate uterus?

OBJECTIVES: To assess the level of agreement between experts in distinguishing between septate and normal/arcuate uterus using their subjective judgment when reviewing the coronal view of the uterus from three-dimensional ultrasound. Another aim was to determine the interobserver reliability and diagnostic test accuracy of three measurements suggested by recent guidelines, using as reference standard the decision made most often by experts (Congenital Uterine Malformation by Experts (CUME)). METHODS: Images of the coronal plane of the uterus from 100 women with suspected fundal internal indentation were anonymized and provided to 15 experts (five clinicians, five surgeons and five sonologists). They were instructed to indicate whether they believed the uterus to be normal/arcuate (defined as normal uterine morphology or not clinically relevant degree of distortion caused by internal indentation) or septate (clinically relevant degree of distortion caused by internal indentation). Two other observers independently measured indentation depth, indentation angle and indentation-to-wall-thickness (I:WT) ratio. The agreement between experts was assessed using kappa, the interobserver reliability was assessed using the concordance correlation coefficient (CCC), the diagnostic test accuracy was assessed using the area under the receiver-operating characteristics curve (AUC) and the best cut-off value was assessed using Youden's index, considering as the reference standard the choice made most often by the experts (CUME). RESULTS: There was good agreement between all experts (kappa, 0.62). There were 18 septate and 82 normal/arcuate uteri according to CUME; European Society of Human Reproduction and Embryology (ESHRE)-European Society for Gynaecological Endoscopy (ESGE) criteria (I:WT ratio > 50%) defined 80 septate and 20 normal/arcuate uteri, while American Society for Reproductive Medicine (ASRM) criteria defined five septate (depth > 15 mm and angle < 90°), 82 normal/arcuate (depth < 10 mm and angle > 90°) and 13 uteri that could not be classified (referred to as the gray-zone). The agreement between ESHRE-ESGE and CUME was 38% (kappa, 0.1); the agreement between ASRM criteria and CUME for septate was 87% (kappa, 0.39), and considering both septate and gray-zone as septate, the agreement was 98% (kappa, 0.93). Among the three measurements, the interobserver reproducibility of indentation depth (CCC, 0.99; 95% CI, 0.98-0.99) was better than both indentation angle (CCC, 0.96; 95% CI, 0.94-0.97) and I:WT ratio (CCC, 0.92; 95% CI, 0.90-0.94). The diagnostic test accuracy of these three measurements using CUME as reference standard was very good, with AUC between 0.96 and 1.00. The best cut-off values for these measurements to define septate uterus were: indentation depth ≥ 10 mm, indentation angle < 140° and I:WT ratio > 110% . CONCLUSIONS: The suggested ESHRE-ESGE cut-off value overestimates the prevalence of septate uterus while that of ASRM underestimates this prevalence, leaving in the gray-zone most of the uteri that experts considered as septate. We recommend considering indentation depth ≥ 10 mm as septate, since the measurement is simple and reliable and this criterion is in agreement with expert opinion. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Optimization of oestradiol assays to improve utility in an in vitro fertilization setting.

Background The measurement of oestradiol is an integral component for the management of ovarian stimulation for in vitro fertilization. Automated immunoassays offer fast assay times and high throughput, with less sensitivity and specificity. The aim of this study is to optimize the oestradiol assay in patients undergoing ovarian stimulation for in vitro fertilization via comparison of oestradiol values obtained using two immunoassays compared with mass spectrometry. Methods Patients undergoing ovarian stimulation were prospectively recruited. Serum samples were analysed with ADVIA Centaur® CP Immunoassay, Abbott Architect i1000® immunoassay and AB Sciex 5500 liquid chromatography-tandem mass spectrometry (LC-MS/MS) systems. Per cent bias was determined for each system to report the average tendency of the values to be larger or smaller than the LC-MS/MS value. Linear regression of total follicular volume and oestradiol was computed. Results The ADVIA Centaur® CP assay had a positive bias of 20% compared with LC-MS/MS, while the Architect i1000® had a non-significant, negative bias of 0.3%. With regression fit, a clear, positive relationship was seen between follicular volume and oestradiol. The Architect i1000® assay had a greater correlation (R2 = 0.46) compared with Centaur® CP (R2 = 0.36), when oestradiol values were >1000 pg/mL (3670 pmol/L). Conclusions The Abbott Architect i1000® oestradiol assay exhibits greater agreement with LC-MS/MS and exhibited better correlation to follicular volume when oestradiol values are >1000 pg/mL (3670 pmol/L), prompting a change in the clinic's oestradiol platform. Attention to assay quality assurance via LC-MS/MS can improve the oestradiol accuracy and permit more informed clinical decisions for improved patient outcomes.

Utilizing pretreatment and fungal incubation to enhance the nutritional value of canola meal.

AIMS: The objective of this study was to determine the optimal pretreatment and fungal strain to reduce glucosinolates (GLS), fibre and residual sugars while increasing the nutritional value of canola meal. METHODS AND RESULTS: Submerged incubation conditions were used to evaluate four pretreatment methods (extrusion, hot water cook, dilute acid and dilute alkali) and three fungal cultures (Aureobasidium pullulans Y-2311-1, Fusarium venenatum NRRL-26139 and Trichoderma reesei NRRL-3653) in hexane-extracted (HE) and cold-pressed (CP) canola meal. CONCLUSIONS: The combination of extrusion pretreatment followed by incubation with T. reesei resulted in the greatest overall improvement to HE canola meal, increasing protein to 51·5%, while reducing NDF, GLS and residual sugars to 18·6%, 17·2 µmol l-1  g-1 and 5% w/w, respectively. Extrusion pretreatment and incubation with F. venenatum performed the best with CP canola meal, resulting in 54·4% protein while reducing NDF, GLS and residual sugars to 11·6%, 6·7 µmol l-1  g-1 and 3·8% w/w respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The work is significant in that it provides a method of reducing GLS (up to 98%) and neutral detergent fibre (up to 65%) while increasing the protein content (up to 45%) of canola meal. This novel pretreatment and submerged incubation process could be used to produce a canola product with higher nutritional value for livestock consumption.

Growth performance of calves fed microbially enhanced soy protein in pelleted starters.

Our objective was to determine effects of feeding calves pelleted starters with microbially enhanced (fungi-treated) soy protein (MSP) in replacement of soybean meal (SBM) with different milk replacers (MR). Thirty-six Holstein calves (2 d old; 24 females, 12 males) in individual hutches were used in a 12-wk randomized complete block design study. Treatments were (1) MSP pellets with MR formulated for accelerated growth (28% crude protein, 18% fat; MSPA), (2) SBM pellets with MR formulated for accelerated growth (SBMA), and (3) MSP pellets with conventional MR (20% crude protein, 20% fat; MSPC). Pellets were similar except for 23% MSP or 23% SBM (dry matter basis). Pellets and water were fed ad libitum throughout the study. Feeding rates of MR on a dry matter basis were 0.37kg twice daily during wk 1, 0.45kg twice daily during wk 2 to 5, and 0.45kg once daily during wk 6. Intakes were recorded daily. Body weights, frame size measurements, and jugular blood samples were collected 2 d every 2 wk at 3 h after the morning feeding. Fecal grab samples were collected 5 times per d for 3 d during wk 12 and then composited by calf for analysis of apparent total-tract digestibility of nutrients using acid detergent insoluble ash as an internal marker. Total and starter pellet dry matter intake were greatest for calves fed SBMA and least for MSPC. Calves had similar average daily gain among treatments, but there was a treatment by week interaction and during the last few weeks of the study calves on MSPC had less body weight compared with MSPA or SBMA. Gain-to-feed ratio was similar among treatments; however, there was a treatment by week interaction. Serum glucose was similar among treatments. Plasma urea nitrogen was greatest for calves fed MSPA and least for MSPC. Plasma concentrations of IGF-1 were greatest for calves fed SBMA. Plasma concentrations of triglycerides were greatest for calves fed MSPC. Plasma concentrations of ß-hydroxybutyrate had a treatment by time interaction. Treatments had similar total-tract dry matter digestibility, but calves fed MSPC had greater crude protein digestibility than SBMA, with MSPA similar to both. Results demonstrated calves fed pelleted starters with MSP had maintained growth performance with less starter intake compared with SBM.

Evaluation of microbially enhanced soybean meal as an alternative to fishmeal in weaned pig diets.

An experimental, microbially enhanced soybean product (MEPRO) was evaluated as a replacement for fishmeal (FM). Assessment of feedstuffs should include estimation of digestibility as well as pig performance and in combination with dietary additives. Digestibility values determined in growing pigs may not apply to nursery pigs; thus, standardized ileal digestibility (SID) of amino acids (AA) in MEPRO and FM were determined using 30±1.6 kg BW ileal-cannulated barrows (n=6) and 9.8±1.2 kg BW barrows (n=37; serial slaughter). Experimental diets included MEPRO, FM and nitrogen free where FM and MEPRO were included as the sole protein source. The SID of AAs was 3% to 5% lower in MEPRO than FM when fed to 30 kg pigs. The SID of arginine and methionine was greater (P<0.05) in MEPRO than FM when fed to 10 kg pigs. The SID of AAs was 12% to 20% lower in FM when fed to 10 v. 30 kg pigs but only 3% to 9% lower in MEPRO. A total of 336 barrows and gilts were weaned at 21 days of age (initial BW=6.1±0.8 kg) and used in a performance trial. Pens of pigs were assigned to one of the six experimental diets (8 pens/diet in two blocks). Treatment diets were fed in Phase I (7 days) and Phase II (14 days) with all pigs fed a common Phase III diet (14 days). Experimental diets included (1) negative control (NEG) containing corn, soybean meal and whey, (2) NEG+acidifier, (3) NEG+FM (POS), (4) POS+acidifier (POS A+), (5) NEG+MEPRO (MEPRO) and (6) MEPRO+acidifier. The FM and MEPRO were included at 7.5% and 5.0% in Phase I and II diets, respectively. Diets were formulated to meet the standard nutrient requirements for weaned pigs. Pig BW and feed disappearance was measured weekly and fecal scores were measured daily for the first 14 days post-weaning as an indicator of post-weaning diarrhea syndrome (PWDS). Performance (BW, daily gain, feed intake and gain : feed) was not significantly different among treatments. Treatment for PWDS occurred on different days in each block. Analysis of fecal score was completed separately by block. Pigs fed the NEG diets had higher (P=0.02) fecal scores than pigs fed the POS diets on days 2 and 3 (block 1) and higher (P<0.05) than pigs fed MEPRO or POS diets and diets with dietary acidifier on days 6 and 3 (block 2). The MEPRO holds promise as an alternative to FM in nursery pig diets.

Evaluation of a bioprocessed soybean meal on nursery pig performance and immune status.

A total of 239 pigs (initial BW 6.56 ± 0.87 kg, 21 d of age) were used in a 35-d study to investigate the effects of fish meal (FM) and spray dried plasma (SDP) in combination with a bioprocessed soybean meal (SBM) on growth performance and immune responses in weaned pigs. Equal numbers of barrows and gilts were randomly allotted to 1 of 4 dietary treatments (10 pens/treatment) according to initial BW and sex: positive control (CON; corn/SBM diet) containing SDP and FM, the CON with bioprocessed SBM replacing FM (BPSBM), the CON with bioprocessed SBM replacing SDP (BPSBM), and the CON with bioprocessed SBM replacing both SDP and FM (BPSBM). Experimental diets were fed in Phase I (d 1-7 post-wean) and II (d 8-21) followed by a common Phase III diet (d 22-35); changes in BW and feed disappearance were determined accordingly. Pigs were sensitized against ovalbumin (OVA) and Candida albicans (CAA) on d 7 and 21. Assessment of immune response was based on lymphocyte proliferation in response to mitogens Concanavalin A and phytohemagglutinin (d 14 post-wean), dermal hypersensitivity to OVA and CAA (% increase in local swelling at 2, 6, 24, and 48 h post-injection) on d 28, and primary and secondary anti-OVA IgG at d 21 and 28, respectively. Pigs fed CON were heavier (P < 0.01) than pigs fed BPSBM and BPSBM, and not different from pigs fed BPSBM, at the end of Phase I and II (6.99, 6.80, 6.52, or 6.60 kg, pooled SEM 0.08, respectively in Phase I or 12.47, 12.18, 11.42, and 11.85 kg, pooled SEM 0.21, in Phase II, respectively). Hypersensitivity to OVA peaked at 2h in pigs fed CON, BPSBM, and BPSBM or peaked at 6h in pigs fed BPSBM (121.4, 165.6, 139.0, and 144.1%, pooled SEM 22.9, respectively, at 2 h and 86.7, 114.5, 95.0, and 156.8%, pooled SEM 29.4, respectively at 6 h). Peak response to CAA occurred at 2h in all groups (42.6, 55.2, 48.2, and 50.6%, pooled SEM 11.9, respectively, in the CON, BPSBM, BPSBM, and BPSBM, respectively). There was no difference in hypersensitivity due to experimental diet at any time point. Secondary anti-OVA IgG was 2-fold lower based on optical density values in pigs fed CON compared with BPSBM and BPSBM (0.78 vs. 1.56 and 1.55 optical density at 405 nm, pooled SEM 0.42, respectively). Dietary treatment did not impact lymphocyte proliferation. The bioprocessed SBM is a suitable alternative for FM and/or SDP in Phase I and II nursery diets based on pig growth. The prolonged hypersensitivity to OVA indicate that bioprocessed SBM may have a positive impact on pig immune function and the 2-fold increase in anti-OVA IgG warrants further investigation on the impact of bioprocessed SBM on pig immune function.

Sequential extrusion-ozone pretreatment of switchgrass and big bluestem.

Pretreatment is one of the biggest challenges in utilizing lignocellulosic feedstocks to meet the mandatory requirements for biofuels around the world. Earlier researchers evaluated extrusion and ozone pretreatment separately and found that sugar recovery can be improved significantly from 15-20 to 40-75 % for different feedstocks. To further improve sugar recoveries, extrusion-ozone sequential pretreatment was explored. Accordingly, optimal extruded switchgrass (176 °C, 155 rpm, 20 % moisture, and 8 mm) and big bluestem (180 °C, 155 rpm, 20 % moisture, and 8 mm) at 25-75 % moisture content were exposed to an ozone flow rate of 37-365 mg/h for 2.5 to 10 min. Pretreated samples were then subjected to enzymatic hydrolysis to determine sugar recovery. Statistical analyses confirmed significant effects of the independent variables and their interactions on sugar recoveries for both feedstocks. Maximum glucose, xylose, and total sugar recovery of 66.4, 82.3, and 70.4 %, respectively, were obtained when a low-moisture (25 %) extruded switchgrass was ozonated for 2.5 min at a flow rate of 37 mg/h. Respectively, this represents increases of 3.42, 5.01, and 3.42 times that of the control. When big bluestem at 25 % moisture was extruded and then ozonated for 2.5 min at a flow rate of 365 mg/h, resulting glucose, xylose, and total sugar recoveries of 90.8, 92.2, and 87.5 %, respectively, were obtained. These represent increases of 4.5, 2.7, and 3.9 times than that of the control. It is also noteworthy that furfural and hydroxymethyl furfural were not detected in any of the pretreatments, and only low levels (0.14-0.18 g/l) of acetic acid were measured. The results show that sequential pretreatment using extrusion and ozone is an efficient way to improve sugar recovery from herbaceous biomass feedstocks.

Sequential extrusion-microwave pretreatment of switchgrass and big bluestem.

Developing an effective and economical biomass pretreatment method is a significant roadblock to meeting the ever growing demand for transportation fuels. Earlier studies with different feedstocks revealed that in the absence of chemicals, neither extrusion nor microwave could be standalone pretreatments. However, there is potential that the advantages of these individual methods can be harnessed in a sequential pretreatment process. Accordingly, switchgrass and big bluestem were extruded and then subject to microwave pretreatment, under optimal conditions that had been separately determined in prior studies. Pretreated biomass was then subject to enzymatic hydrolysis to understand the effectiveness of the sequential pretreatment on sugar recovery and generation of fermentation inhibitors. Statistical analysis confirmed that moisture content, microwave power level, and exposure time (and their interactions) had significant influence on sugar recovery. Sequential pretreatment of switchgrass (25% moisture, 450W and 2.5min) resulted in a maximum glucose, xylose, and total sugar recovery of 52.6%, 75.5%, and 59.2%, respectively. This was higher by 1.27 and 2.71, 1.21 and 4.60, and 1.25 and 2.87 times compared to extrusion alone and the unpretreated control, respectively. The same sequential pretreatment conditions achieved maximum glucose, xylose, and total sugar recovery of 83.2%, 92.1%, and 68.1%, respectively, for big bluestem. This was 1.14 and 4.1, 1.18 and 2.7, and 1.20 and 3.0 times higher than extrusion alone and the unpretreated control, respectively. This sequential pretreatment process did not aggravate acetic acid formation over levels observed with the individual pretreatments. Furthermore, furfural, HMF, and formic acid were not detected in any of the treatments. Although the sequential pretreatment process enhanced sugar recovery without increasing the levels of potential fermentation inhibitors, the increased energy input for the microwave treatment may not be economical.

Extrusion pretreatment of pine wood chips.

Pretreatment is the first step to open up lignocellulose structure in the conversion of biomass to biofuels. Extrusion can be a viable pretreatment method due to its ability to simultaneously expose biomass to a range of disruptive conditions in a continuous flow process. Extruder screw speed, barrel temperature, and feedstock moisture content are important factors that can influence sugar recovery from biomass. Hence, the current study was undertaken to investigate the effects of these parameters on extrusion pretreatment of pine wood chips. Pine wood chip at 25, 35, and 45 % wb moisture content were pretreated at various barrel temperatures (100, 140, and 180 °C) and screw speeds (100, 150, and 200 rpm) using a screw with compression ratios of 3:1. The pretreated pine wood chips were subjected to standard enzymatic hydrolysis followed by sugar and byproducts quantification. Statistical analyses revealed the existence of significant differences in sugar recovery due to independent variables based on comparing the mean of main effects and interaction effects. Pine wood chips pretreated at a screw speed of 150 rpm and a barrel temperature of 180 °C with a moisture content of 25 % resulted in a maximum cellulose, hemicellulose, and total sugar recoveries of 65.8, 65.6, and 66.1 %, respectively, which was about 6.7, 7.9, and 6.8 fold higher than the control (unpretreated pine chips). Furthermore, potential fermentation inhibitors such as furfural, hydroxyl methyl furfural, and acetic acid were not found in any of the treatment combinations.

Profiling transient daytime peaks in urban air pollutants: city centre traffic hotspot versus urban background concentrations.

Averaged annual and daily limit values imposed for the protection of human health from urban air pollutants fail to address the potential influence of variations in personal exposure during the day, even though asthma attacks, myocardial ischaemia, and other effects have been linked to highly transient rises in atmospheric contamination. There is, therefore, a general need for the wider availability of information on hourly variations in urban pollutant levels, as well as on spatial differences between urban background and localised contamination hotspots. We compare data from urban background and traffic hotspot monitoring stations over a 12-month period to demonstrate the hourly flux of air pollutants, notably NO(x), SO(2) and PM(10), and illustrate the effects of controlling influences, such as traffic movements and exotic dust intrusions. Our methodology focuses on daytime transient peaks, which occur when much of the urban population is awake and active, and when striking increases in pollutant concentrations are especially common at traffic hotspots. Greater emphasis on the daily reality of transient peaks in urban atmospheric pollutants offers a higher degree of spatial and temporal data resolution for use in epidemiological studies.

Conversion of volatile fatty acids into polyhydroxyalkanoate by Ralstonia eutropha.

AIMS: The aims of this study were to optimize condensed corn solubles (CCS) as a medium for growth of Ralstonia eutropha and to determine the effects of individual volatile fatty acids (VFAs) on polyhydroxyalkanoate (PHA) production. METHODS AND RESULTS: A CCS medium of concentration 240 g l(-1) with a carbon : nitrogen ratio of 50 : 1 was developed as the optimal medium. Cultures were grown in 1-l aerated flasks at 250 rev min(-1) at 30 degrees C for 120 h. Comparable growth rates were observed in CCS vs a defined medium. At 48 h, VFAs were fed individually at different levels. Optimal levels of all the acids were determined to maximize PHA production. An overall comparison of the VFAs indicated that butyric and propionic acids provided the best results. CONCLUSION: An optimized CCS medium supported growth of R. eutropha. Butyric and propionic acids were the most efficient carbon sources to maximize PHA production when added at the 5 g l(-1) level. SIGNIFICANCE AND IMPACT OF THE STUDY: The study shows that a byproduct of ethanol industry can be effectively used as a low cost medium for PHA production, thus partly reducing the cost of commercialization of biopolymers.

Spatial and temporal variations in inhalable CuZnPb aerosols within the Mexico City pollution plume.

We report on the CuPbZn content of PM10 and PM2.5 samples collected from three sites (urban T0, suburban T1 and rural T2) during the Mexico City MILAGRO campaign of March 2006. Daytime city centre concentrations of summation operator CuZnPb(PM10) were much higher (T0 > 450 ng m(-3)) than at the suburban site (T1 < 200 ng m(-3)). Rural site (T2) summation operator CuZnPb(PM10) concentrations exceeded 50 ng m(-3) when influenced by the megacity plume but dropped to 10 ng m(-3) during clean northerly winds. Nocturnal metal concentrations more than doubled at T0, as pollutants became trapped in the nightly inversion layer, but decreased at the rural site. Transient spikes in concentrations of different metals, e.g. a "copper event" at T0 (CuPM10 281 ng m(-3)) and "zinc event" at T1 (ZnPM10 1481 ng m(-3)) on the night of March 7-8, demonstrate how industrial pollution sources produce localised chemical inhomogeneities in the city atmosphere. Most metal aerosols are <2.5 microm and SEM study demonstrates the dominance of Fe, Ti, Ba, Cu, Pb and Zn (and lesser Sn, Mo, Sb, W, Ni, V, As, Bi) in metalliferous particles that have shapes including spherical condensates, efflorescent CuZnClS particles, cindery Zn, and Cu wire. Metal aerosol concentrations do not change in concert with PM10 mass, which is more influenced by wind resuspension than industrial emissions. Metalliferous particles can induce cell damage, and PM composition is probably more important than PM mass, with respect to negative health effects, so that better monitoring and control of industrial emissions would likely produce significant improvements in air quality.

Rhodospirillum rubrum: utilization of condensed corn solubles for poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) production.

AIMS: This study sought to develop a less expensive medium for growth of the polyhydroxyalkanoate-producing bacterium Rhodospirillum rubrum from the ethanol production coproduct, condensed corn solubles (CCS). METHODS AND RESULTS: Small-scale trials using R. rubrum were performed in aerated or anaerobic stoppered serum bottles filled with media. The CCS (240 g l(-1)) achieved a maximum cell density and growth rate comparable with the defined supplemented malate-ammonium medium (mSMN) or tryptic soy broth. Microaerophilic solubles medium cultures exhibited significantly higher maximum cell densities and growth rates than did strictly anaerobic cultures; while illumination, nickel or biotin addition had no effect. Growth of R. rubrum in a pH controlled bioreactor was significantly better in CCS (240 g l(-1)) than in mSMN medium and supported production of 0.36% (cell dry weight) poly-(3-hydroxybutyrate-Co-3-hydroxyvalerate) after 24 h. CONCLUSIONS: A CCS medium was devised that supported R. rubrum growth for biopolymer production as effective as the defined medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that a more economical medium can be developed for biopolymer production using a low value coproduct from ethanol production. The impact is that this inexpensive solubles medium may make it more economical to produce the biopolymer on a commercial scale.

Oocyte biology and genetics revelations from polar bodies.

The enormous volume of the fertilized egg is attributable to the suppression of cleavage during oocyte growth and the unequal cleavages during the first and second meiotic divisions. The two products of these divisions are the diminutive polar bodies (PB), which contain a redundant set of chromosomes/chromatids plus cytoplasmic organelles. The PB have strictly limited but differential life spans; while viable they possess the genetic potential to support normal embryonic development after transfer to a cytoplast. In addition to the theoretical possibility of using this non-cloning technique to generate more embryos, polar bodies can be used for genetic testing. By cytogenetic analysis of both PB using fluorescent in-situ hybridization (FISH) or chromosome painting, partial or full chromosomal status in the oocyte can be predicted; this approach finds particular application for women of advanced reproductive age as well as with maternally inherited translocations and single gene defects. By studying both of the PB, potential problems of interpretation arising from allele dropout can be reduced; a heterozygous first polar body provides the least ambiguous result. Mitochondria segregate randomly during meiotic cleavages providing an opportunity also to use the PB to screen for mitochondrial mutations and deletions. Thus, the PB can serve useful diagnostic purposes, especially where pre-fertilization screening or avoidance of embryo biopsy is desirable.

Delaying the initiation of progesterone supplementation results in decreased pregnancy rates after in vitro fertilization: a randomized, prospective study.

OBJECTIVE: To compare IVF outcome between two protocols for luteal phase supplementation, one beginning on day 3 after oocyte retrieval and the other beginning on day 6 after retrieval. DESIGN: Prospective, randomized study. SETTING: University-based assisted reproductive technology center. PATIENT(S): One hundred twenty-six consecutive patients undergoing IVF between January and July 2000. INTERVENTION(S): Patients were randomized to begin luteal phase support using vaginal progesterone beginning either on day 3 after oocyte retrieval or on day 6 after oocyte retrieval. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates and implantation rates. RESULT(S): All patients randomized underwent transfer. There were no differences in age, oocytes retrieved, or embryos transferred between the two groups. Those patients receiving luteal phase support with progesterone beginning on day 6 after retrieval had a significantly lower clinical pregnancy rate per transfer compared with those beginning support on day 3 after retrieval (44.8% vs. 61.0%, respectively). This difference in pregnancy rates was greater in those patients undergoing a luteal gonadotropin releasing hormone (GnRH) agonist down-regulation protocol (47.5% vs. 71.4%, day 6 vs. day 3, respectively). Beginning support on day 6 also significantly decreased implantation rates in the GnRH agonist group (21.0% vs. 34.0%, day 6 vs. day 3, respectively). CONCLUSION(S): Pregnancy rates are significantly decreased by initiating luteal-phase progesterone supplementation on day 6 after oocyte retrieval during in vitro fertilization cycles.

High FSH:LH ratio and low LH levels in basal cycle day 3: impact on follicular development and IVF outcome.

PURPOSE: To examine the impact of low basal cycle day 3 serum LH levels or a high FSH:LH ratio on IVF results. METHODS: A homogeneous group of patients was analyzed as identified by normal basal cycle of follicle stimulating hormone (FSH), Luteinizing hormone (LH), and estradiol (E2) levels. High responders (high LH:FSH ratio) and low responders (high FSH or E2 levels, and women > or = 42 years of age) were excluded from analysis. Only cycles stimulated with a combination of a GnRHa (luteal suppression) and pure FSH were studied. RESULTS: Patients with low basal LH levels (< 3 mIU/mL) did not differ significantly from controls in terms of response to controlled ovarian hyperstimulation but there was a clear trend toward poorer implantation and clinical pregnancy rates. On the other hand, patients with a high FSH:LH ratio (> 3) had significantly fewer mature oocytes aspirated, and lower implantation and clinical pregnancy rates than patients with gonadotropin ratio < or = 3. These negative effects were evident in the presence of normal basal FSH levels and after adequate matching of female's age and number of embryos transferred. CONCLUSIONS: These studies highlight a negative impact of a basal cycle high FSH:LH ratio (and possibly low LH levels) on follicular development and oocyte quality in these patients subjected to pituitary down-regulation followed by pure FSH administration. A high FSH:LH ratio may be therefore used as an early biomarker of poor ovarian response.

Characterization of telomerase activity in the human oocyte and preimplantation embryo.

Telomerase, a ribonucleoprotein, has been described as an essential component of highly proliferative cells as it stabilizes the telomeres and avoids cellular senescence. The objective of this study was to modify the polymerase chain reaction-based telomeric repeat amplification protocol to detect telomerase activity in the single cell and to characterize the activity expressed in the human oocyte through to the blastocyst stage embryo. A comparative evaluation of telomerase activity and developmental stage was conducted using discarded or donated human oocytes and embryos. Telomerase activity was detected in all developmental stages evaluated from immature oocytes through to blastocyst stage embryos. Immature oocytes and blastocysts had similar levels of telomerase activity; however, both groups had significantly (P < 0.05) higher activity than zygote through to pre-morula stage embryos. Seventy-five thawed zygotes were cultured to day 3, biopsied by removing 1-2 cells, and the biopsied embryos were cultured to blastocyst stage. There was no difference (P < 0.05) in telomerase activity between cells biopsied from embryos that reached the blastocyst stage and cells from those that arrested in growth. This study has shown that human oocytes through to blastocyst stage embryos express telomerase activity, but that the level of telomerase activity in biopsied blastomeres, of the day 3 cleavage stage embryo, is not predictive of embryonic growth potential.