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.

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.

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.

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.

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.

Drought stress has transgenerational effects on soybean seed germination and seedling vigor.

Effects of environmental stressors on the parent may be transmitted to the F1 generation of plants that support global food, oil, and energy production for humans and animals. This study was conducted to determine if the effects of drought stress on parental soybean plants are transmitted to the F1 generation. The germination and seedling vigor of F1 soybean whose maternal parents, Asgrow AG5332 and Progeny P5333RY, were exposed to soil moisture stress, that is, 100, 80, 60, 40, and 20% replacement of evapotranspiration (ET) during reproductive growth, were evaluated under controlled conditions. Pooled over cultivars, effects of soil moisture stress on the parents caused a reduction in the seed germination rate, maximum seed germination, and overall seedling performance in the F1 generation. The effect of soil moisture stress on the parent environment induced seed quality that carried on the F1 generation seed gemination and seedling traits under optimum conditions and further exasperated when exposed to increasing levels of drought stress. Results indicate that seed weight and storage reserve are key factors positively associated with germination traits and seedling growth. Our data confirm that the effects of soil moisture stress on soybean are transferable, causing reduced germination, seedling vigor, and seed quality in the F1 generation. Therefore, optimal water supply during soybean seed formation period may be beneficial for seed producers in terms of optimizing seed quality and vigor characteristics of commodity seed.

Evaluating rice for salinity using pot-culture provides a systematic tolerance assessment at the seedling stage.

BACKGROUND: Rice (Oryza sativa L.) is one of the major staple food crops consumed globally. However, rice production is severely affected by high salinity levels, particularly at the seedling stage. A good solution would be the development of an efficient screening methodology to identify genotypes possessing genes for salt tolerance. RESULT: A new salinity tolerance screening technique using rice seedlings in pot-culture was tested. This method controls soil heterogeneity by using pure sand as a growth medium and minimizes unexpected extreme weather conditions with a movable shelter. Seventy-four rice genotypes were screened at three salinity treatments including high salt stress (electrical conductivity (EC) 12 dSm- 1), moderate salt stress (EC 6 dSm- 1), and control (no salt stress), imposed 1 week after emergence. Several shoot and root morpho-physiological traits were measured at 37 days after sowing. A wide range of variability was observed among genotypes for measured traits with root traits being identified as the best descriptors for tolerance to salt stress conditions. Salt stress response indices (SSRI) were used to classify the 74 rice genotypes; 7 genotypes (9.46%) were identified as salt sensitive, 27 (36.48%) each as low and moderately salt tolerant, and 13 (17.57%) as highly salt tolerant. Genotypes FED 473 and IR85427 were identified as the most salt tolerant and salt sensitive, respectively. These results were further confirmed by principal component analysis (PCA) for accuracy and reliability. CONCLUSION: Although tolerant genotypes still need to be confirmed in field studies and tolerance mechanisms identified at the molecular level, information gained from this study could help rice breeders and other scientists to accelerate breeding by selecting appropriate donor parents, progenies and potential genotypes at early growth stages necessary for salinity tolerance research.

Soybean seed physiology, quality, and chemical composition under soil moisture stress.

Soybean seed quality is often determined by its constituents which are important to sustain overall nutritional aspects. The objective of this study was to investigate the effects of soil moisture stress during reproductive stage on seed quality and composition. Plants were subjected to five levels of soil moisture stresses at flowering, and yield and quality traits were examined at maturity. Seed protein, palmitic and linoleic acids, sucrose, raffinose, stachyose, N, P, K, and Ca significantly decreased whereas oil, stearic, oleic and linolenic acids, Fe, Mg, Zn, Cu, and B increased in response to soil moisture deficiency. The relationship between seed protein and oil was negatively correlated. The changes in seed constituents could be due to changes in nutrient accumulation and partitioning in soybean seeds under water stress. This information suggests the requirement of adequate soil moisture during flowering and seed formation stages to obtain the higher nutritional value of soybean seeds.

Interactive effects on CO2, drought, and ultraviolet-B radiation on maize growth and development.

Crop growth and development are highly responsive to global climate change components such as elevated carbon dioxide (CO2), drought, and ultraviolet-B (UV-B) radiation. Plant tolerance to these environmental stresses comprises its genetic potential, physiological changes, metabolism, and signaling pathways. An inclusive understanding of morphological, physiological, and biochemical responses to these abiotic stresses is imperative for the development of stress tolerant varieties for future environments. The objectives of this study were to characterize the changes in vegetative and physiological traits in maize hybrids in their response to multiple environmental factors of (CO2) [400 and 750µmolmol(-1) (+(CO2)], irrigation treatments based evapotranspiration (ET) [100 and 50% (-ET)], and UV-B radiation [0 and 10kJm(-2)d(-1) (+UV-B)] and to identify the multiple stress tolerant hybrids aid in mitigating projected climate change for shaping future agriculture. Six maize hybrids (P1498, DKC 65-81, N75H-GTA, P1319, DKC 66-97, and N77P-3111) with known drought tolerance variability were grown in eight sunlit, controlled environment chambers in which control treatment consisted of 400µmolmol(-1) [CO2], 100% ET-based irrigation, and 0kJ UV-B. Plants grown at +UV-B alone or combination with 50% ET produced shorter plants and smaller leaf area while elevated CO2 treatments ameliorated the damaging effects of drought and higher UV-B levels on maize hybrids. Plant height, leaf area, total dry matter chlorophyll, carotenoids, and net photosynthesis measured were increased in response to CO2 enrichment. Total stress response index (TSRI) for each hybrid, developed from the cumulative sum of response indices of vegetative and physiological parameters, varied among the maize hybrids. The hybrids were classified as tolerant (P1498), intermediate (DKC 65-81, N75H-GTA, N77P-3111) and sensitive (P1319 and DKC 66-97) to multiple environmental stresses. The positive correlation between TSRI and vegetative and physiological index developed in this study demonstrates that a combination of vegetative and physiological traits is an effective screening tool to identify germplasm best suited to cope with future changing climates. Furthermore, the tolerant hybrids identified in this study indicate that the possibility of cultivar selection for enhanced agronomic performance and stability in a water limited environment with higher UV-B, anticipated to occur in future climates.

Discovery of a Cyclic Boronic Acid ß-Lactamase Inhibitor (RPX7009) with Utility vs Class A Serine Carbapenemases.

The increasing dissemination of carbapenemases in Gram-negative bacteria has threatened the clinical usefulness of the ß-lactam class of antimicrobials. A program was initiated to discover a new series of serine ß-lactamase inhibitors containing a boronic acid pharmacophore, with the goal of finding a potent inhibitor of serine carbapenemase enzymes that are currently compromising the utility of the carbapenem class of antibacterials. Potential lead structures were screened in silico by modeling into the active sites of key serine ß-lactamases. Promising candidate molecules were synthesized and evaluated in biochemical and whole-cell assays. Inhibitors were identified with potent inhibition of serine carbapenemases, particularly the Klebsiella pneumoniae carbapenemase (KPC), with no inhibition of mammalian serine proteases. Studies in vitro and in vivo show that RPX7009 (9f) is a broad-spectrum inhibitor, notably restoring the activity of carbapenems against KPC-producing strains. Combined with a carbapenem, 9f is a promising product for the treatment of multidrug resistant Gram-negative bacteria.

Antiviral efficacy upon administration of a HepDirect prodrug of 2'-C-methylcytidine to hepatitis C virus-infected chimpanzees.

Hepatitis C virus (HCV) infects an estimated 170 million individuals worldwide, and the current standard of care, a combination of pegylated interferon alpha and ribavirin, is efficacious in achieving sustained viral response in ~50% of treated patients. Novel therapies under investigation include the use of nucleoside analog inhibitors of the viral RNA-dependent RNA polymerase. NM283, a 3'-valyl ester prodrug of 2'-C-methylcytidine, has demonstrated antiviral efficacy in HCV-infected patients (N. Afdhal et al., J. Hepatol. 46[Suppl. 1]:S5, 2007; N. Afdhal et al., J. Hepatol. 44[Suppl. 2]:S19, 2006). One approach to increase the antiviral efficacy of 2'-C-methylcytidine is to increase the concentration of the active inhibitory species, the 5'-triphosphate, in infected hepatocytes. HepDirect prodrug technology can increase intracellular concentrations of a nucleoside triphosphate in hepatocytes by introducing the nucleoside monophosphate into the cell, bypassing the initial kinase step that is often rate limiting. Screening for 2'-C-methylcytidine triphosphate levels in rat liver after oral dosing identified 1-[3,5-difluorophenyl]-1,3-propandiol as an efficient prodrug modification. To determine antiviral efficacy in vivo, the prodrug was administered separately via oral and intravenous dosing to two HCV-infected chimpanzees. Circulating viral loads declined by ~1.4 log(10) IU/ml and by >3.6 log(10) IU/ml after oral and intravenous dosing, respectively. The viral loads rebounded after the end of dosing to predose levels. The results indicate that a robust antiviral response can be achieved upon administration of the prodrug.