Laser-induced breakdown spectroscopy (LIBS) of animal fat (lard): Efficient sample preparation for onsite analysis and influence of sample temperature on the signal intensity and plasma parameters.

We report an efficient sample preparation method (freezing) for onsite fat and meat analysis via a specially designed thermoelectric cooling and temperature-controlling system. This investigation also focused on the effect of phase change on the sensitivity and reproducibility of LIBS emission signals and plasma parameters. The plasma emissions of animal fats (lard) were recorded when the sample was frozen (-2 °C), fluid (15 °C), and in a liquid state (37 °C) with a thermoelectric cooling system. At each temperature, the plasma emissions were acquired at laser pulse energy from 50 to 300 mJ and detector gate delay (DGD) from 0.5 to 5 µs. With increasing sample temperature, the DGD, where the optical emission intensity reached a maximum, decreased. At a laser pulse energy of 200 mJ and a sample temperature of -2 °C, the emission signals increased fourfold, the signal-to-noise ratio (SNR) improved tenfold, and the self-absorption in the emission lines decreased significantly. The repeatability of the emission signals and plasma parameters of frozen and liquid fat samples was determined using the relative standard deviation (RSD) of Se I (473.08 nm) and K I (766.48 nm) emission lines. The RSDs of the emission signals improved from 40 to 18 % and 37 to 16 %, whereas the shot-to-shot RSDs of the electron temperature and electron number density get improved from 11 to 6 % and 12 to 6.8 %, respectively.

Impact of soil moisture stress during the silk emergence and grain-filling in maize.

Suboptimal soil moisture during the growing season often limits maize growth and yield. However, the growth stage-specific responses of maize to soil moisture regimes have not been thoroughly investigated. This study investigated the response of maize to five different soil moisture regimes, that are, 0.25, 0.20, 0.15, 0.10, and 0.05 m3 m-3 volumetric water content (VWC), during flowering and grain-filling stages. Sub-optimal soil moisture at the flowering and grain-filling stages reduced ear leaf stomatal conductance by 73 and 64%, respectively. An increase in stress severity caused significant reductions in ear leaf chlorophyll content and greenness-associated vegetation indices across growth stages. Fourteen days of soil moisture stress during flowering delayed silk emergence, reduced silk length (19%), and silk fresh weight (34%). Furthermore, sub-optimal soil moisture caused a significant reduction in both kernel number (53%) and weight (54%). Soil moisture stress at the flowering had a direct impact on kernel number and an indirect effect on kernel weight. During grain-filling, disruption of ear leaf physiology resulted in a 34% decrease in kernel weight and a 43% decrease in kernel number. Unlike grain-filling, treatments at the flowering significantly reduced kernel starch (3%) and increased protein by 29%. These findings suggest that developing reproductive stage stress-tolerant hybrids with improved resilience to soil moisture stress could help reduce the yield gap between irrigated and rainfed maize.

Phenological stage and vegetation index for predicting corn yield under rainfed environments.

Uncrewed aerial systems (UASs) provide high temporal and spatial resolution information for crop health monitoring and informed management decisions to improve yields. However, traditional in-season yield prediction methodologies are often inconsistent and inaccurate due to variations in soil types and environmental factors. This study aimed to identify the best phenological stage and vegetation index (VI) for estimating corn yield under rainfed conditions. Multispectral images were collected over three years (2020-2022) during the corn growing season and over fifty VIs were analyzed. In the three-year period, thirty-one VIs exhibited significant correlations (r ≥ 0.7) with yield. Sixteen VIs were significantly correlated with the yield at least for two years, and five VIs had a significant correlation with the yield for all three years. A strong correlation with yield was achieved by combining red, red edge, and near infrared-based indices. Further, combined correlation and random forest an alyses between yield and VIs led to the identification of consistent and highest predictive power VIs for corn yield prediction. Among them, leaf chlorophyll index, Medium Resolution Imaging Spectrometer (MERIS) terrestrial chlorophyll index and modified normalized difference at 705 were the most consistent predictors of corn yield when recorded around the reproductive stage (R1). This study demonstrated the dynamic nature of canopy reflectance and the importance of considering growth stages, and environmental conditions for accurate corn yield prediction.

Posterior Ethmomaxillary Cells: Anatomical Variation to be Considered in Endoscopic Sinus Surgery.

To identify the posterior ethmomaxillary (PEM) cells on computed tomographic scans for complete sinus clearance during endoscopic sinus surgery to prevent disease recurrence. Prospective single institution study. MAA ENT Hospitals Pvt. Ltd., Hyderabad. 350 patients were selected for the study. Computed tomographic scans of these patients were performed suffering from chronic rhinosinusitis and requiring endoscopic sinus surgery (primary or revision cases). The scans were evaluated and presence of PEM cells was noted. These findings were co-related intra-operatively and above-mentioned cells were opened. In revision cases, these cells were left unopened in previous cases and were cleared in present surgery and the patients were followed up for recurrence. A total number of 350 CT scans of nose and para-nasal sinus were studied. There were 176 males and 174 were females. The presence of PEM cells was 11.42% with bilaterality in 80%. In revision cases, it was 23%. PEM cells act as hidden areas in the para-nasal sinus and if not identified and cleared, it acts as a nidus for disease recurrence and cause for failure of surgery. Identification of PEM is important during surgery for complete disease clearance. As very little information exists in the current literature, we are submitting this study to enlighten Rhinologist regarding PEM cells.

Effects of fine grinding on mid-infrared spectroscopic analysis of plant leaf nutrient content.

Fourier transform mid infrared (FT-MIR) spectroscopy combined with modeling techniques has been studied as a useful tool for multivariate chemical analysis in agricultural research. A drawback of this method is the sample preparation requirement, in which samples must be dried and fine ground for accurate model calibrations. For research involving large sample sets, this may dramatically increase the time and cost of analysis. This study investigates the effect of fine grinding on model performance using leaf tissue from a variety of crop species. Dried leaf samples (N = 300) from various environmental conditions were obtained with data on 11 nutrients measured using chemical methods. The samples were scanned with attenuated total reflectance (ATR) and diffuse reflectance (DRIFT) FT-MIR techniques. Scanning was repeated after fine grinding for 2, 5, and 10 min. The spectra were analyzed for the 11 nutrients using partial least squares regression with a 75%/25% split for calibration and validation and repeated for 50 iterations. All analytes except for boron, iron, and zinc were well-modeled (average R2 > 0.7), with higher R2 values on ATR spectra. The 5 min level of fine grinding was found to be most optimal considering overall model performance and sample preparation time.

Interference of Nanoparticulates in seed invigoration of Green gram.

In the present study, the impact of four metal/metal oxide nanoparticles (NPs) viz.Ag, ZnO),ZVI and TiO2 on physiological seed quality attributes of green gram (Vigna radiata) were evaluated. The synthesized NPs characterized and evaluated the germination percentage, vigour indices and physiological responses like catalase and peroxidase activities (seed quality parameters) of fresh, naturally aged and fresh accelerated aged seed lots of green gram. In naturally aged seeds, zinc oxide-NPs (1000 mg kg-1) treated seeds showed 14.96% higher germination percentage, 24.81% higher vigour index I and (3696) and 33.33% higher vigour index II than the controls. The treated seeds with ZnO-NPs (1000 mg kg-1) under fresh accelerated aged conditions resulted in higher than 15.15% of germination percentage, 23.61% of vigour index I and 24.11% of vigour index II over controls. Moreover, ZnO-NPs treated naturally aged seeds showed lower electrical conductivity (EC) of 20.10 µ S cm-1g-1 than the control (26.60 µ S cm-1 g-1). Pertinent to catalase enzyme activity, ZnO-NPs (1000 mg kg-1) treated naturally aged seed lots resulted in 356.89 µmol H2O2 mg-1 min-1 activity, 216.05 µmol H2O2 mg-1 min-1 activity in fresh accelerated aged seed lots.. Similarly, ZnO-NPs (1000 mg kg-1) enhanced peroxidase enzyme activity in naturally aged seed lots (3.21 µg/FW/10 min) than control (0.72 µg/FW/10 min) that depicts 63.35% of increased enzyme activity. The present results showcases the ZnO-NPs as potent nano-priming agents in maintaining the seed quality parameters that ultimately establish better crop stand and field performance.

Resilience of soybean cultivars to drought stress during flowering and early-seed setting stages.

Drought stress during the reproductive stage and declining soybean yield potential raise concerns about yield loss and economic return. In this study, ten cultivars were characterized for 20 traits to identify reproductive stage (R1-R6) drought-tolerant soybean. Drought stress resulted in a marked reduction (17%) in pollen germination. The reduced stomatal conductance coupled with high canopy temperature resulted in reduced seed number (45%) and seed weight (35%). Drought stress followed by rehydration increased the hundred seed weight at the compensation of seed number. Further, soybean oil decreased, protein increased, and cultivars responded differently under drought compared to control. In general, cultivars with high tolerance scores for yield displayed lower tolerance scores for quality content and vice versa. Among ten cultivars, LS5009XS and G4620RX showed maximum stress tolerance scores for seed number and seed weight. The observed variability in leaf reflectance properties and their relationship with physiological or yield components suggested that leaf-level sensing information can be used for differentiating drought-sensitive soybean cultivars from tolerant ones. The study led to the identification of drought-resilient cultivars/promising traits which can be exploited in breeding to develop multi-stress tolerant cultivars.

Proteomic analysis response of rice (Oryza sativa) leaves to ultraviolet-B radiation stress.

Rice (Oryza sativa) is a human staple food and serves as a model organism for genetic and molecular studies. Few studies have been conducted to determine the effects of ultraviolet-B (UV-B) stress on rice. UV-B stress triggers morphological and physiological changes in plants. However, the underlying mechanisms governing these integrated responses are unknown. In this study, we conducted a proteomic response of rice leaves to UV-B stress using two-dimensional gel electrophoresis and identified the selected proteins by mass spectrometry analysis. Four levels of daily biologically effective UV-B radiation intensities were imposed to determine changes in protein accumulation in response to UV-B stress: 0 (control), 5, 10, and 15 kJ m-2 d-1in two cultivars, i.e., IR6 and REX. To mimic the natural environment, we conducted this experiment in Sunlit Soil-Plant-Atmosphere-Research (SPAR) chambers. Among the identified proteins, 11% of differentially expressed proteins were found in both cultivars. In the Rex cultivar, only 45% of proteins are differentially expressed, while only 27.5% were expressed in IR6. The results indicate that REX is more affected by UV-B stress than IR6 cultivars. The identified protein TSJT1 (spot 16) in both cultivars plays a crucial role in plant growth and development during stress treatment. Additionally, we found that UV-B stress altered many antioxidant enzymes associated with redox homeostasis and cell defense response. Another enzyme, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), has been identified as spot 15, which plays an essential role in glycolysis and cellular energy production. Another vital protein identified is glycosyl hydrolase (GH) as spot 9, which catalyzes the hydrolysis of glycosidic bonds in cell wall polymers and significantly affects cell wall architecture. Some identified proteins are related to photosynthesis, protein biosynthesis, signal transduction, and stress response. The findings of our study provide new insights into understanding how rice plants are tailored to UV-B stress via modulating the expression of UV-B responsive proteins, which will help develop superior rice breeds in the future to combat UV-B stress. Data are available via ProteomeXchange with identifier PXD032163.

Effects of drought and heat stresses during reproductive stage on pollen germination, yield, and leaf reflectance properties in maize (Zea mays L.).

Drought and heat stresses are the major abiotic stress factors detrimental to maize (Zea mays L.) production. Much attention has been directed toward plant responses to heat or drought stress. However, maize reproductive stage responses to combined heat and drought remain less explored. Therefore, this study aimed to quantify the impact of optimum daytime (30°C, control) and warmer daytime temperatures (35°C, heat stress) on pollen germination, morpho-physiology, and yield potential using two maize genotypes ("Mo17" and "B73") under contrasting soil moisture content, that is, 100% and 40% irrigation during flowering. Pollen germination of both genotypes decreased under combined stresses (42%), followed by heat stress (30%) and drought stress (19%). Stomatal conductance and transpiration were comparable between control and heat stress but significantly decreased under combined stresses (83% and 72%) and drought stress (52% and 47%) compared with the control. Genotype "Mo17" reduced its green leaf area to minimize the water loss, which appears to be one of the adaptive strategies of "Mo17" under stress conditions. The leaf reflectance of both genotypes varied across treatments. Vegetation indices associated with pigments (chlorophyll index of green, chlorophyll index of red edge, and carotenoid index) and plant health (normalized difference red-edge index) were found to be highly sensitive to drought and combined stressors than heat stress. Combined drought and heat stresses caused a significant reduction in yield and yield components in both Mo17 (49%) and B73 (86%) genotypes. The harvest index of genotype "B73" was extremely low, indicating poor partitioning efficiency. At least when it comes to "B73," the cause of yield reduction appears to be the result of reduced sink number rather than the pollen and source size. To the best of our awareness, this is the first study that showed how the leaf-level spectra, yield, and quality parameters respond to the short duration of independent and combined stresses during flowering in inbred maize. Further studies are required to validate the responses of potential traits involving diverse maize genotypes under field conditions. This study suggests the need to develop maize with improved tolerance to combined stresses to sustain production under increasing temperatures and low rainfall conditions.

Low- and High-Temperature Phenotypic Diversity of Brassica carinata Genotypes for Early-Season Growth and Development.

Temperature is a major abiotic stress factor limiting plant growth and development during the early developmental stage. Information on carinata (Brassica carinata A. Braun) traits response to low and high temperatures is necessary for breeding or selecting genotypes suited for specific ecoregions, which is limited. In the present study, 12 carinata genotypes were evaluated under low (17/09°C), optimum (22/14°C), and high (27/19°C) day/night temperatures at the early developmental stage. This study quantified temperature effects on several physiological and morphological characteristics of 12-advanced carinata lines. High-temperature plants decreased (15%) the accumulation of flavonoids and increased the nitrogen balance index by 25%. Low-temperature treatment significantly inhibited the aboveground (plant height, leaf area, number, and shoot weight) and root (length, surface area, and weight) traits. Across all genotypes, the shoot weight decreased by 55% and the root weight by 49% under low temperature. On the other hand, the maximum proportion of biomass was partitioned to roots under low temperature than at the high temperature. A poor relationship (r 2 = 0.09) was found between low- and high-temperature indices, indicating differences in trait responses and tolerance mechanisms. AX17004 and AX17009 with higher root to shoot ratios might be suitable for late planting windows or regions with low-temperature spells. The two genotypes (AX17015 and AX17005) accumulated higher biomass under low- and high-temperature treatments can be used for planting in later summer or early winter. The identified low- and high-temperature stress-tolerant carinata genotypes could be a valuable resource for increasing stress tolerance during the early developmental stage.

Morpho-Physiological, Yield, and Transgenerational Seed Germination Responses of Soybean to Temperature.

Temperature is the primary factor affecting the morpho-physiological, developmental, and yield attributes of soybean. Despite several temperature and soybean studies, functional relationships between temperature and soybean physiology and yield components are limited. An experiment was conducted to determine the optimum temperature for soybean gas exchange and yield components using indeterminate (Asgrow AG5332, AG) and determinate (Progeny P5333 RY, PR) growth habit cultivars. Plants grown outdoors were exposed to 5 day/night temperature treatments, 21/13, 25/17, 29/21, 33/25, and 37°C/29°C, from flowering to maturity using the sunlit plant growth chambers. Significant temperature and cultivar differences were recorded among all measured parameters. Gas exchange parameters declined with increasing temperature treatments during the mid-pod filling stage, and quadratic functions best described the response. The optimum temperature for soybean pod weight, number, and seed number was higher for AG than PR, indicating greater high-temperature tolerance. Soybean exposed to warmer parental temperature (37°C/29°C) during pod filling decreased significantly the transgenerational seed germination when incubated at 18, 28, and 38°C. Our findings suggest that the impact of temperature during soybean development is transferable. The warmer temperature has adverse transgenerational effects on seed germination ability. Thus, developing soybean genotypes tolerant to high temperatures will help growers to produce high-yielding and quality beans. The quantified temperature, soybean physiology, and yield components-dependent functional algorithms would be helpful to develop adaptation strategies to offset the impacts of extreme temperature events associated with future climate change.

A Novel Modified-Vista Technique With Connective Tissue Graft in the Treatment of Gingival Recession: A Case Report.

INTRODUCTION: The increasing esthetic demands from patients have required that clinicians sharpen their skills and adopt newer and more novel techniques to satisfy these demands. In periodontal therapy, periodontal plastic surgery poses a substantial challenge to the clinician, both because it is technique sensitive and also because it includes a wide array of procedures and their variations. Conventional plastic procedures have provided satisfactory results in the treatment of gingival recession but there is, presently, a greater need for more advanced procedures that cause less surgical morbidity as also provide improved results. Minimally invasive surgery has been harnessed in periodontics for this purpose and has been extensively used in multiple indications, including root coverage. Modified-Vestibular Incision Supra-periosteal Tunnel Access (M-VISTA) applies the principles of minimally invasive surgery to provide satisfactory results in root coverage. CASE PRESENTATION: A 28-year-old male patient presented with multiple Miller Class I/II buccal recession defects and associated high labial frenum on right maxillary anterior teeth. M-VISTA technique allowed coronal repositioning of gingival margin, which was then stabilized by orthodontics bracket assisted suturing technique. The V-shaped incision in the region of the frenal attachment allowed for conducting both the M-VISTA and frenectomy techniques simultaneously. Connective tissue graft was inserted in the supra-periosteal tunnel and vertical incision was sutured. Complete root coverage was achieved and maintained at 9 months with excellent esthetic outcomes. CONCLUSIONS: The use of connective tissue graft along with M-VISTA technique allows the clinician to successfully treat multiple recession defects.

Sweetpotato cultivars responses to interactive effects of warming, drought, and elevated carbon dioxide.

Plants are sensitive to changes projected in climates, such as elevated carbon dioxide (eCO2), high temperature (T), and drought stress (DS), which affect crop growth, development, and yield. These stresses, either alone or in combination, affect all aspects of sweetpotato plant growth and development, including storage root development and yield. We tested three sweetpotato cultivars (Beauregard, Hatteras, and LA1188) responses to eight treatments (Control, DS, T, eCO2, DS + T, T + eCO2, DS + eCO2, DS + T + eCO2). All treatments were imposed 36 days after transplanting (DAP) and continued for 47 days. Treatments substantially affected gas exchange, photosynthetic pigments, growth, and storage root components. Cultivars differed considerably for many of the measured parameters. The most significant negative impact of DS was recorded for the shoot and root weights. The combination of DS + T had a significant negative effect on storage root parameters. eCO2 alleviated some of the damaging effects of DS and high T in sweetpotato. For instance, eCO2 alone or combined with DS increased the storage root weights by 22% or 42% across all three cultivars, respectively. Based on the stress response index, cultivar "Hatteras" was most tolerant to individual and interactive stresses, and "LA 1188" was sensitive. Our findings suggest that eCO2 negates the negative impact of T or DS on the growth and yield of sweetpotato. We identified a set of individual and interactive stress-tolerant traits that can help select stress cultivars or breed new lines for future environments.

Selection of QPX7831, an Orally Bioavailable Prodrug of Boronic Acid ß-Lactamase Inhibitor QPX7728.

In recognition of the need for effective oral therapies to treat Gram-negative bacterial infections, efforts were directed toward identifying an oral prodrug of ß-lactamase inhibitor clinical candidate QPX7728. Seventeen prodrugs were synthesized; key properties investigated were rates of cleavage to the active form in vitro, pharmacokinetics across species, and crystallinity. Compound 5-Na (QPX7831 Sodium) emerged with optimal properties across all key attributes.

Developing Functional Relationships between Soil Waterlogging and Corn Shoot and Root Growth and Development.

Short- and long-term waterlogging conditions impact crop growth and development, preventing crops from reaching their true genetic potential. Two experiments were conducted using a pot-culture facility to better understand soil waterlogging impacts on corn growth and development. Two corn hybrids were grown in 2017 and 2018 under ambient sunlight and temperature conditions. Waterlogging durations of 0, 2, 4, 6, 8, 10, 12, and 14 days were imposed at the V2 growth stage. Morphological (growth and development) and pigment estimation data were collected 15 days after treatments were imposed, 23 days after sowing. As waterlogging was imposed, soil oxygen rapidly decreased until reaching zero in about 8-10 days; upon the termination of the treatments, the oxygen levels recovered to the level of the 0 days treatment within 2 days. Whole-plant dry weight declined as the waterlogging duration increased, and after 2 days of waterlogging, a 44% and 27% decline was observed in experiments 1 and 2, respectively. Leaf area and root volume showed an exponential decay similar to the leaf and root dry weight. Leaf number and plant height were the least sensitive measured parameters and decreased linearly in both experiments. Root forks were the most sensitive parameter after 14 days of waterlogging in both experiments, declining by 83% and 80% in experiments 1 and 2, respectively. The data from this study improve our understanding of how corn plants react to increasing durations of waterlogging. In addition, the functional relationships generated from this study could enhance current corn simulation models for field applications.

Yield, Physiological Performance, and Phytochemistry of Basil (Ocimum basilicum L.) under Temperature Stress and Elevated CO2 Concentrations.

Early season sowing is one of the methods for avoiding yield loss for basil due to high temperatures. However, basil could be exposed to sub-optimal temperatures by planting it earlier in the season. Thus, an experiment was conducted that examines how temperature changes and carbon dioxide (CO2) levels affect basil growth, development, and phytonutrient concentrations in a controlled environment. The experiment simulated temperature stress, low (20/12 °C), and high (38/30 °C), under ambient (420 ppm) and elevated (720 ppm) CO2 concentrations. Low-temperature stress prompted the rapid closure of stomata resulting in a 21% decline in net photosynthesis. Chlorophylls and carotenoids decreased when elevated CO2 interacted with low-temperature stress. Basil exhibited an increase in stomatal conductance, intercellular CO2 concentration, apparent quantum yield, maximum photosystem II efficiency, and maximum net photosynthesis rate when subjected to high-temperature stress. Under elevated CO2, increasing the growth temperature from 30/22 °C to 38/30 °C markedly increased the antioxidants content of basil. Taken together, the evidence from this research recommends that varying the growth temperature of basil plants can significantly affect the growth and development rates compared to increasing the CO2 concentrations, which mitigates the adverse effects of temperature stress.

Novel Surgical Approach for Root Coverage Using Coronally Advanced Flap in Conjunction With Transposition of Periosteal Pedicle Graft.

INTRODUCTION: Gingival recession is a complex phenomenon with multifactorial etiology. It is defined as the apical migration of the soft tissue margin beyond the cemento-enamel junction, thereby exposing the root surface. It results in the destruction of both soft and hard tissues. CASE PRESENTATION: Three patients with buccal gingival recession defects underwent surgical treatment consisting of transposition of a periosteal pedicle in conjunction with the coronally advanced flap (CAF) technique. As the cambium layer of the periosteum has greater osteoblastic potential than the fibrous layer, this study considered juxtaposing of the cambium layer directly onto the denuded root surface. A 9-month review demonstrated satisfactory: root coverage; gain in clinical attachment, reduction in probing depth; and increase in width of keratinized gingiva with a good color match and minimal scarring. CONCLUSION: Within the limits of the study, this CAF technique, in conjunction with the cambium layer of periosteum, showed a significant amount of root coverage.

The duration of embryo culture after mouse IVF differentially affects cardiovascular and metabolic health in male offspring.

STUDY QUESTION: Do the long-term health outcomes following IVF differ depending upon the duration of embryo culture before transfer? SUMMARY ANSWER: Using a mouse model, we demonstrate that in male but not female offspring, adverse cardiovascular (CV) health was more likely with prolonged culture to the blastocyst stage, but metabolic dysfunction was more likely if embryo transfer (ET) occurred at the early cleavage stage. WHAT IS KNOWN ALREADY: ART associate with increased risk of adverse CV and metabolic health in offspring, and these findings have been confirmed in animal models in the absence of parental infertility issues. It is unclear which specific ART treatments may cause these risks. There is increasing use of blastocyst, versus cleavage-stage, transfer in clinical ART which does not appear to impair perinatal health of children born, but the longer-term health implications are unknown. STUDY DESIGN, SIZE, DURATION: Five mouse groups were generated comprising: (i) natural mating (NM)-naturally mated, non-superovulated and undisturbed gestation; (ii) IV-ET-2Cell-in-vivo derived two-cell embryos collected from superovulated mothers, with immediate ET to recipients; (iii) IVF-ET-2Cell-IVF generated embryos, from oocytes from superovulated mothers, cultured to the two-cell stage before ET to recipients; (iv) IV-ET-BL-in-vivo derived blastocysts collected from superovulated mothers, with immediate ET to recipients; (v) IVF-ET-BL-IVF generated embryos, from oocytes from superovulated mothers, cultured to the blastocyst stage before ET to recipients. Both male and female offspring were analysed for growth, CV and metabolic markers of health. There were 8-13 litters generated for each group for analyses; postnatal data were analysed by multilevel random effects regression to take account of between-mother and within-mother variation and litter size. PARTICIPANTS/MATERIALS, SETTINGS, METHODS: C57/BL6 female mice (3-4 weeks old) were used for oocyte production; CBA males for sperm with human tubal fluid medium were used for IVF. Embryos were transferred (ET) to MF1 pseudo-pregnant recipients at the two-cell stage or cultured in synthetic oviductal medium enriched with potassium medium to the blastocyst stage before ET. Control in-vivo embryos from C57BL6 × CBA matings were collected and immediately transferred at the two-cell or blastocyst stage. Postnatal assays included growth rate up to 27 weeks; systolic blood pressure (SBP) at 9, 15 and 21 weeks; lung and serum angiotensin-converting enzyme (ACE) activity at time of cull (27 weeks); glucose tolerance test (GTT; 27 weeks); basal glucose and insulin levels (27 weeks); and lipid accumulation in liver cryosections using Oil Red O imaging (27 weeks). MAIN RESULTS AND THE ROLE OF CHANCE: Blastocysts formed by IVF developed at a slower rate and comprised fewer cells that in-vivo generated blastocysts without culture (P < 0.05). Postnatal growth rate was increased in all four experimental treatments compared with NM group (P < 0.05). SBP, serum and lung ACE and heart/body weight were higher in IVF-ET-BL versus IVF-ET-2Cell males (P < 0.05) and higher than in other treatment groups, with SBP and lung ACE positively correlated (P < 0.05). Glucose handling (GTT AUC) was poorer and basal insulin levels were higher in IVF-ET-2Cell males than in IVF-ET-BL (P < 0.05) with the glucose:insulin ratio more negatively correlated with body weight in IVF-ET-2Cell males than in other groups. Liver/body weight and liver lipid droplet diameter and density in IVF-ET-2Cell males were higher than in IVF-ET-BL males (P < 0.05). IVF groups had poorer health characteristics than their in-vivo control groups, indicating that outcomes were not caused specifically by background techniques (superovulation, ET). No consistent health effects from duration of culture were identified in female offspring. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Results from experimental animal models cannot be extrapolated to humans. Nevertheless, they are valuable to develop conceptual models, in this case, in the absence of confounding parental infertility, in assessing the safety of ART manipulations. WIDER IMPLICATIONS OF THE FINDINGS: The study indicates that longer duration of embryo culture after IVF up to blastocyst before ET leads to increased dysfunction of CV health in males compared with IVF and shorter cleavage-stage ET. However, the metabolic health of male offspring was poorer after shorter versus longer culture duration. This distinction indicates that the origin of CV and metabolic health phenotypes after ART may be different. The poorer metabolic health of males after cleavage-stage ET coincides with embryonic genome activation occurring at the time of ET. STUDY FUNDING/COMPETING INTEREST(S): This work was supported through the European Union FP7-CP-FP Epihealth programme (278418) and FP7-PEOPLE-2012-ITN EpiHealthNet programme (317146) to T.P.F., the Biotechnology and Biological Sciences Research Council (BBSRC) (BB/F007450/1) to T.P.F., and the Saudi government, University of Jeddah and King Abdulaziz University to A.A. The authors have no conflicts of interest to declare.

Discovery of Cyclic Boronic Acid QPX7728, an Ultrabroad-Spectrum Inhibitor of Serine and Metallo-ß-lactamases.

Despite major advances in the ß-lactamase inhibitor field, certain enzymes remain refractory to inhibition by agents recently introduced. Most important among these are the class B (metallo) enzyme NDM-1 of Enterobacteriaceae and the class D (OXA) enzymes of Acinetobacter baumannii. Continuing the boronic acid program that led to vaborbactam, efforts were directed toward expanding the spectrum to allow treatment of a wider range of organisms. Through key structural modifications of a bicyclic lead, stepwise gains in spectrum of inhibition were achieved, ultimately resulting in QPX7728 (35). This compound displays a remarkably broad spectrum of inhibition, including class B and class D enzymes, and is little affected by porin modifications and efflux. Compound 35 is a promising agent for use in combination with a ß-lactam antibiotic for the treatment of a wide range of multidrug resistant Gram-negative bacterial infections, by both intravenous and oral administration.