Reduced Growth Rate Research Articles

Climatic drivers and tree growth in a key production species: The case of Cryptomeria japonica (Thunb. ex L.f.) D.Don in the Azores archipelago

Forest management plays a crucial role in preserving and enhancing the delivery of ecosystem services, whereas dendrochronological methods can play a significant role in this regard. In the Azores archipelago, with a temperate oceanic climate, with low thermal amplitude, and mild and relatively wet Summers, Cryptomeria japonica (Thunb. ex L.f.) D.Don is the main timber production species. It has been extensively planted for timber production and erosion control, being the mainstay of the regional forestry chain, with a rotation period of 30 years. Although dendrochronological studies have targeted this species elsewhere, this dendrochronological study in the Azores aimed to better understand its climate-growth relationships in the archipelago. For this purpose, we sampled 140 trees, in a total of 361 wood cores samples. Following standard dendrochronological methods, we obtained four site chronologies from different volcanic complexes in São Miguel island. We used a stepwise modelling approach, with Random Forest, Principal Component Analysis and Generalized Linear Models. Our results suggest that despite year-round precipitation in the Azores, C. japonica is adversely affected by warm Winter and Summers, while on site SC it benefits from these conditions. As previously observed, precipitation may be overshadowed by temperature-driven mechanisms in C. japonica. Our spatial analysis resulted into climate-growth associations with similar directions for all site chronologies. In the future, higher Winters and Summer temperatures could lead to increased water stress and reduced growth rates. This should be considered when projecting the future distribution and productivity of C. japonica forests under different climate change scenarios. To ensure the long-term survival of this economically important tree species, adaptation responses should include genetic and conservation measures. Our findings provide baseline information for defining management approaches for this strategic species.

Probing the Effects of Retinoblastoma Binding Protein 6 (RBBP6) Knockdown on the Sensitivity of Cisplatin in Cervical Cancer Cells.

Cervical cancer is a major cause of death in women despite the advancement of current treatment modalities. The conventional therapeutic agent, cisplatin (CCDP), is the standard treatment for CC; however, resistance often develops due to the cancer's heterogeneity. Therefore, a detailed elucidation of the specific molecular mechanisms driving CC is crucial for the development of targeted therapeutic strategies. Retinoblastoma binding protein 6 (RBBP6) is a potential biomarker associated with cell proliferation and is upregulated in cervical cancer sites, exhibiting apoptosis and dysregulated p53 expression. Furthermore, RBBP6 has been demonstrated to sensitize cancer cells to radiation and certain chemotherapeutic agents by regulating the Bcl-2 gene, thus suggesting a crosstalk among RBBP6/p53/BCL-2 oncogenic signatures. The present study, therefore, investigated the relationship between cisplatin and RBBP6 expression in CC cells. Herein, we first explored bioinformatics simulations and identified that the RBBP6/p53/BCL-2 signaling pathway is overexpressed and correlated with CC. For further analysis, we explored the Genomics of Drug Sensitivity in Cancer (GDSC) and found that most of the CC cell lines are sensitive to CCDP. To validate these findings, RBBP6 was silenced in HeLa and Vero cells using RNAi technology, followed by measurement of wild-type p53 and Bcl-2 at the mRNA level using qPCR. Cells co-treated with cisplatin and siRBBP6 were subsequently analyzed for apoptosis induction and real-time growth monitoring using flow cytometry and the xCELLigence system, respectively. Cancer cells in the co-treatment group showed a reduction in apoptosis compared to the cisplatin-treated group. Moreover, the real-time growth monitoring revealed a reduced growth rate in RBBP6 knockdown cells treated with cisplatin. Although wild-type p53 remained unchanged in the co-treatment group of cancer cells, Bcl-2 was completely repressed, suggesting that RBBP6 is necessary for sensitizing cervical cancer cells to cisplatin treatment by downregulating Bcl-2. The Vero cell population, which served as a non-cancerous control cell line in this study, remained viable following treatment with both siRBBP6 and cisplatin. Findings from this study suggest that RBBP6 expression promotes cisplatin sensitivity in HeLa cells through Bcl-2 downregulation. Knockdown of RBBP6 limits apoptosis induction and delays cell growth inhibition in response to cisplatin. The knowledge obtained here has the potential to help improve cisplatin efficacy through personalized administration based on the expression profile of RBBP6 among individual patients.

Identification of factors limiting the allotopic production of the Cox2 subunit of yeast cytochrome c oxidase.

Mitochondrial genes can be artificially relocalized in the nuclear genome in a process known as allotopic expression, such is the case of the mitochondrial cox2 gene, encoding subunit II of cytochrome c oxidase (CcO). In yeast, cox2 can be allotopically expressed and is able to restore respiratory growth of a cox2-null mutant if the Cox2 subunit carries the W56R substitution within the first transmembrane stretch. However, the COX2W56R strain exhibits reduced growth rates and lower steady-state CcO levels when compared to wild-type yeast. Here, we investigated the impact of overexpressing selected candidate genes predicted to enhance internalization of the allotopic Cox2W56R precursor into mitochondria. The overproduction of Cox20, Oxa1, and Pse1 facilitated Cox2W56R precursor internalization, improving the respiratory growth of the COX2W56R strain. Overproducing TIM22 components had a limited effect on Cox2W56R import, while overproducing TIM23-related components showed a negative effect. We further explored the role of the Mgr2 subunit within the TIM23 translocator in the import process by deleting and overexpressing the MGR2 gene. Our findings indicate that Mgr2 is instrumental in modulating the TIM23 translocon to correctly sort Cox2W56R. We propose a biogenesis pathway followed by the allotopically produced Cox2 subunit based on the participation of the 2 different structural/functional forms of the TIM23 translocon, TIM23MOTOR and TIM23SORT, that must follow a concerted and sequential mode of action to insert Cox2W56R into the inner mitochondrial membrane in the correct Nout-Cout topology.

Structural and functional analyses of nematode-derived antimicrobial peptides support the occurrence of direct mechanisms of worm-microbiota interactions

The complex relationships between gastrointestinal (GI) nematodes and the host gut microbiota have been implicated in key aspects of helminth disease and infection outcomes. Nevertheless, the direct and indirect mechanisms governing these interactions are, thus far, largely unknown. In this proof-of-concept study, we demonstrate that the excretory-secretory products (ESPs) and extracellular vesicles (EVs) of key GI nematodes contain peptides that, when recombinantly expressed, exert antimicrobial activity in vitro against Bacillus subtilis. In particular, using time-lapse microfluidics microscopy, we demonstrate that exposure of B. subtilis to a recombinant saposin-domain containing peptide from the ‘brown stomach worm’, Teladorsagia circumcincta, and a metridin-like ShK toxin from the ‘barber’s pole worm’, Haemonchus contortus, results in cell lysis and significantly reduced growth rates. Data from this study support the hypothesis that GI nematodes may modulate the composition of the vertebrate gut microbiota directly via the secretion of antimicrobial peptides, and pave the way for future investigations aimed at deciphering the impact of such changes on the pathophysiology of GI helminth infection and disease.

Long-term evaluation of pasture production, seasonality, and variability: An application of the DairyMod pasture model for three tropical species

Adoption of improved pastures coupled with intensified management provide quality pastures in adequate quantities and thus improve livestock productivity. While pasture modelling is imperative for exploring the performance of newer pastures, models are little used for long-term simulations of multiple tropical pastures (genotype), under varying soil, climate (environment) and pasture production systems (management). We applied the DairyMod, a biophysical model to simulate the long-term pasture production of Brachiaria ruziziensis x B. decumbens x B. brizantha ‘Brachiaria Mulato II’ (BM), Megathyrsus maximus ‘Gatton Panic’ (GP), and Chloris gayana ‘Rhodes grass cv. Reclaimer’ (RR) across major dairying regions of Sri Lanka under different management scenarios and characterize the long-term pasture growth, seasonality and spatial variability, and possible implications for dairying in Sri Lanka. Simulations of three pasture species were carried out for 16 locations (8 dry (DZ), 5 intermediate (IZ), and 3 wet zone (WZ)) over 30 years (1980–2010). Three pasture management scenarios simulated were; 1) potential pasture production system under non-limiting N and irrigation (Yp) 2) rainfed pasture production system under non-limiting N fertilizer (Yw), and 3) rainfed pasture production system under current nitrogen (N) fertilizer rate (Ya). Statistical techniques were used to identify the long-term growth rates, variability, and trends in pasture production. The long-term pasture production varied greatly among climate, species, and management scenarios. Overall, the Ya showed a seasonal cycle following the rainfall pattern, with a reduction in growth rates in dry seasons (May–September). Pasture growth rates were greater in GP at Ya, and BM at Yw and Yp while RR showed the lowest growth rate at all times. Variability of pasture growth was high in DZ (May–September) and RR has the lowest growth variability. The Yw increased the growth rate (doubled) while the Yp substantially increased (nearly tripled) the growth rate and growth pattern producing less variable pastures. Simulated growth rates suggest that GP in low-input and BM in high-input farming areas would be more suitable. Our study suggested that the BM, GP, and RR are edaphic-climatologically fit for major dairying regions in Sri Lanka and the appropriate fertilizer and irrigation management can greatly increase the herbage accumulation and availability of year-round pastures. While this study offers valuable insights, the species-specific growth pattern, growth variability, yield potential under different managements and the possible implications for herbage quality need to be sensibly considered when selecting the appropriate species.

A supermultiplier demand-led growth accounting analysis applied to the Spanish economy (1998–2019)

In this paper, we analyse the demand-led determinants of Spanish economic growth from 1998–2019. We apply the supermultiplier demand-led growth accounting methodology by Freitas/Dweck (2013) with two modifications: First, we incorporate consumption out of public transfers, following Haluska et al. (2021) and Haluska (2023). Second, we incorporate consumption out of public wages as a source of autonomous demand, theoretically suggested by Serrano/Pimentel (2019). Our demand-led growth decomposition highlights: (i) public demand and exports as important stable growth drivers, and a decreasing supermultiplier that reduces growth rates; (ii) the indirect effect of a real estate boom in the economic expansion of 1998–2008 caused by increasing public revenues and opening space to the expansion of public demand; and (iii) the incapacity of exports to lead the recovery alone, as the latter started only with the resumption of the public and private demand.

Antimicrobial and antibiofilm effect of cannabinoids from Cannabis sativa against methicillin-resistant Staphylococcus aureus (MRSA) causing bovine mastitis.

Antimicrobial resistance (AMR) poses a serious threat to human, animal, and plant health on a global scale. Search and elimination techniques should be used to effectively counter the spread of methicillin-resistant Staphylococcus aureus (MRSA) infections. With only a few novel drugs in clinical development, the quest for plant-based alternatives to prevent the spread of antibiotic resistance among bacteria has accelerated. Treatment of MRSA infections is challenging owing to rapidly emerging resistance mechanisms coupled with their protective biofilms. In the present research, we examined the antibacterial properties of ten plant-derived ethanolic leaf extracts. The most effective ethanolic leaf extract against MRSA in decreasing order of zone of inhibition, Cannabis sativa L. > Syzygium cumini > Murraya koenigii > Eucalyptus sp. > while Aloe barbadensis, Azadirachta indica, had very little impact. Mangifera indica, Curcuma longa, Tinospora cordifolia, and Carica papaya did not exhibit inhibitory effects against MRSA; hence, Cannabis was selected for further experimental study. The minimal inhibitory concentration (MIC) of Cannabis sativa L. extract was 0.25mgml-1 with 86% mortality. At a sub-MIC dosage of 0.125mgml-1, the biofilm formation was reduced by 71%. The two major cannabinoids detected were cannabidiol and delta-9-tetrahydrocannabinol (Δ9-THC), which were majorly attributed to substantial inhibitory action against MRSA. The time-kill kinetics demonstrated a bactericidal action at 4 MIC over an 8-20-h time window with a 90% reduction in growth rate. The results from SEM, and light microscopy Giemsa staining revealed a reduction in cells in the treated group with increased AKP activity, indicating bacterial cell membrane breakdown. These findings suggested cannabinoids may be a promising alternative to antibiotic therapy for bovine biofilm-associated MRSA.

The Gene vepN Regulated by Global Regulatory Factor veA That Affects Aflatoxin Production, Morphological Development and Pathogenicity in Aspergillus flavus.

Velvet (VeA), a light-regulated protein that shuttles between the cytoplasm and the nucleus, serves as a key global regulator of secondary metabolism in various Aspergillus species and plays a pivotal role in controlling multiple developmental processes. The gene vepN was chosen for further investigation through CHIP-seq analysis due to significant alterations in its interaction with VeA under varying conditions. This gene (AFLA_006970) contains a Septin-type guanine nucleotide-binding (G) domain, which has not been previously reported in Aspergillus flavus (A. flavus). The functional role of vepN in A. flavus was elucidated through the creation of a gene knockout mutant and a gene overexpression strain using a well-established dual-crossover recombinational technique. A comparison between the wild type (WT) and the ΔvepN mutant revealed distinct differences in morphology, reproductive capacity, colonization efficiency, and aflatoxin production. The mutant displayed reduced growth rate; dispersion of conidial heads; impaired cell wall integrity; and decreased sclerotia formation, colonization capacity, and aflatoxin levels. Notably, ΔvepN exhibited complete growth inhibition under specific stress conditions, highlighting the essential role of vepN in A. flavus. This study provides evidence that vepN positively influences aflatoxin production, morphological development, and pathogenicity in A. flavus.

Heavy metal tolerance and accumulation in the Brassica species (Brassica chinensis var. parachinensis and Brassica rapa L.): A pot experiment

This study delves into the heavy metal tolerance and accumulation capabilities of Brassica chinensis var. parachinensis (B. chinensis) and Brassica rapa L. (B. rapa) in a pot experiment, specifically focusing on cadmium (Cd), chromium (Cr) and lead (Pb). Agricultural topsoils were spiked with varying concentrations of these heavy metals (0 mg/kg, 75 mg/kg, 150 mg/kg, 225 mg/kg and 300 mg/kg) for each element. The experiment involved cultivating 15 pots each of B. chinensis and B. rapa over 60 days. Results indicated that both Brassica species experienced delayed germination, with B. chinensis exhibiting a significant drop in germination percentage to 53 % at the highest concentration (300 mg/kg), while B. rapa showed a tendency for an increased germination percentage of up to 80 % at elevated metal concentrations; however, these differences were not statistically significant. Both B. chinensis and B. rapa demonstrated a stable decline in growth rate from 0.05 cm/day to 0.04 cm/day with increasing heavy metal concentrations, and the he reduction in relative growth rate was significant at the highest concentration compared to the control. The stress tolerance index revealed a significant decrease in plant heights for B. chinensis, in contrast to the stable performance of B. rapa, showcasing the tolerance of B. rapa to toxic conditions. Despite insignificant differences in fresh biomass due to metal treatments, B. chinensis consistently yielded higher biomass, yet it had a lower edible index due to its higher root biomass. Leaf areas increased significantly in both species at higher soil treatments, while root lengths remained unchanged, suggesting their resilience to elevated heavy metal concentrations. Analysis of plant tissues (leaves, stems and roots) using ICP-OES revealed that B. rapa accumulated the highest Cd concentration (864 mg/kg), whereas B. chinensis accumulated the highest Pb concentration (953 mg/kg) in root parts. Both species significantly accumulated Cr in roots, demonstrating a sequestration mechanism. These findings suggest that both species, particularly, B. rapa possess strong tolerance and accumulation capabilities for non-essential heavy metals, making them potential hyperaccumulators for green remediation techniques in toxic soil environments. Understanding the molecular mechanisms driving these responses and validating phytoremediation potential in real-world scenarios is essential for developing sustainable soil management practices.

Temperature-dependent toxicity of fluoxetine alters the thermal plasticity of marine diatoms

Anthropogenic activities have led to the emergence of pharmaceutical pollution in marine ecosystems, posing a significant threat to biodiversity in conjunction with global climate change. While the ecotoxicity of human drugs on aquatic organisms is increasingly recognized, their interactions with environmental factors, such as temperature, remain understudied. This research investigates the physiological effects of the selective serotonin reuptake inhibitor (SSRI), fluoxetine, on two diatom species, Phaeodactylum tricornutum and Thalassiosira weissflogii. Results demonstrate that fluoxetine significantly reduces growth rate and biomass production, concurrently affecting pigment contents and the thermal performance curve (TPC) of the diatoms. Fluoxetine reduces the synthesis of chlorophyll a (Chl a) and carotenoid (Car), indicating inhibition of photosynthesis and photoprotection. Furthermore, fluoxetine decreases the maximum growth rate (μmax) while increasing the optimum temperature (Topt) in both species, suggesting an altered thermal plasticity. This shift is attributed to the observed decrease in the inhibition rate of fluoxetine with rising temperatures. These findings emphasize the physiological impacts and ecological implications of fluoxetine on phytoplankton and underscore the significance of considering interactions between multiple environmental drivers when accessing the ecotoxicity of potential pollutants. The present study provides insights into crucial considerations for evaluating the impacts of pharmaceutical pollution on marine primary producers.

Antibiotic resistance begets more resistance: chromosomal resistance mutations mitigate fitness costs conferred by multi-resistant clinical plasmids.

Plasmids are the primary vectors of horizontal transfer of antibiotic resistance genes among bacteria. Previous studies have shown that the spread and maintenance of plasmids among bacterial populations depend on the genetic makeup of both the plasmid and the host bacterium. Antibiotic resistance can also be acquired through mutations in the bacterial chromosome, which not only confer resistance but also result in changes in bacterial physiology and typically a reduction in fitness. However, it is unclear whether chromosomal resistance mutations affect the interaction between plasmids and the host bacteria. To address this question, we introduced 13 clinical plasmids into a susceptible Escherichia coli strain and three different congenic mutants that were resistant to nitrofurantoin (ΔnfsAB), ciprofloxacin (gyrA, S83L), and streptomycin (rpsL, K42N) and determined how the plasmids affected the exponential growth rates of the host in glucose minimal media. We find that though plasmids confer costs on the susceptible strains, those costs are fully mitigated in the three resistant mutants. In several cases, this results in a competitive advantage of the resistant strains over the susceptible strain when both carry the same plasmid and are grown in the absence of antibiotics. Our results suggest that bacteria carrying chromosomal mutations for antibiotic resistance could be a better reservoir for resistance plasmids, thereby driving the evolution of multi-drug resistance.IMPORTANCEPlasmids have led to the rampant spread of antibiotic resistance genes globally. Plasmids often carry antibiotic resistance genes and other genes needed for its maintenance and spread, which typically confer a fitness cost on the host cell observed as a reduced growth rate. Resistance is also acquired via chromosomal mutations, and similar to plasmids they also reduce bacterial fitness. However, we do not know whether resistance mutations affect the bacterial ability to carry plasmids. Here, we introduced 13 multi-resistant clinical plasmids into a susceptible and three different resistant E. coli strains and found that most of these plasmids do confer fitness cost on susceptible cells, but these costs disappear in the resistant strains which often lead to fitness advantage for the resistant strains in the absence of antibiotic selection. Our results imply that already resistant bacteria are a more favorable reservoir for multi-resistant plasmids, promoting the ascendance of multi-resistant bacteria.

The influence of the mass transport regime on laser chemical vapor deposited carbon fibers

Laser Chemical Vapor Deposition (LCVD) is a process through which fibers are additively deposited by the dissociation and deposition of a precursor gas into the solid phase under a translating laser focal point. In the hyperbaric pressure conditions, LCVD has two identified reaction regimes. The first being limited by surface reaction kinetics (SRK) and the second being limited by mass transport (MT). This article examines the effects of mass transport (MT) limitations on carbon fiber growth using either ethane (C2H6) or ethylene (C2H4). Both hydrocarbon precursors reveal a growth rate reduction when transitioning from SRK-to-MT to meet the no-slip condition in fluid dynamics. This transition correlates to a fiber microstructure change from a core shell morphology with a graphitic structure (SRK) to a nodular morphology with an amorphous structure (MT). The fiber fracture stress is generally found to be higher in the MT than SRK regime because of the reduction of easily cleaved graphitic planes in the MT fibers. Collectively, these results demonstrate the importance of the MT limited growth regime and how fiber characteristics can be manipulated when grown in this regime.

Costs of being a diet generalist for the protist predator Dictyostelium discoideum

Consumers range from specialists that feed on few resources to generalists that feed on many. Generalism has the clear advantage of having more resources to exploit, but the costs that limit generalism are less clear. We explore two understudied costs of generalism in a generalist amoeba predator, Dictyostelium discoideum, feeding on naturally co-occurring bacterial prey. Both involve costs of combining prey that are suitable on their own. First, amoebas exhibit a reduction in growth rate when they switched to one species of prey bacteria from another compared to controls that experience only the second prey. The effect was consistent across all six tested species of bacteria. These switching costs typically disappear within a day, indicating adjustment to new prey bacteria. This suggests that these costs are physiological. Second, amoebas usually grow more slowly on mixtures of prey bacteria compared to the expectation based on their growth on single prey. There were clear mixing costs in three of the six tested prey mixtures, and none showed significant mixing benefits. These results support the idea that, although amoebas can consume a variety of prey, they must use partially different methods and thus must pay costs to handle multiple prey, either sequentially or simultaneously.

On the validity of the Tada stress intensity factor solution for the single edge notch tension specimen with pinned ends

The validity of the Tada stress intensity factor (KTada) for pinned-ends single edge notch tension [SEN(T)] specimens is assessed via a combined experimental-modeling approach. Analysis of fatigue crack growth rate reductions during constant-ΔKTada loading demonstrates that specific combinations of alloy stiffness, geometry, and loading result in the true K deviating below KTada. Geometrically non-linear, 3-dimensional finite element calculations confirm mild-to-strong influences of these parameters, which are not captured by KTada. Most existing pinned SEN(T) data are found to use parameters where KTada is not significantly reduced, but the present results underscore the need for a more broadly applicable K solution.

Biosurfactant-Assisted Phytoremediation of Diesel-Contaminated Soil by Three Different Legume Species

This study aims to assess the impact of HydroBreak PLUS biosurfactant on the phytoremediation of diesel-contaminated soil by three legume plant species: Medicago sativa, Lotus corniculatus, and Melilotus albus. Legumes were grown in soil contaminated with diesel (4.0 g kg−1, 6.0 g kg−1) for 90 days, and the changes in soil diesel and nutrient concentrations, plant growth, and physiological parameters were measured. Diesel negatively affected the biomass production of all legumes, though the reduction in growth rate was observed only in L. corniculatus and M. albus. L. corniculatus had the highest diesel removal rate of 93%, M. albus had the lowest of 87.9%, and unplanted treatments had significantly lower diesel removal rates (up to 66.5%). The biosurfactant mitigated diesel-induced reduction in plant shoot and root weight and an increase in L. corniculatus root biomass (24.2%) were observed at 4.0 g kg−1 diesel treatment. The use of biosurfactant accelerated diesel removal from the soil, though the effect was diesel soil concentration and plant species-dependent. In unplanted treatments, the diesel removal rates increased by 16.4% and 6.9% in the treatments with 4 and 6 mg kg−1, respectively. The effect of biosurfactants on diesel removal by plants was less pronounced and reached 4.6% and 3.2% in the treatments with 4 and 6 mg kg−1, respectively. The study revealed that the phytoremediation efficiency could not be directly linked to plant physiological parameters as only M. sativa changes in plant growth corresponded well with photosystem II performance. Implementation of legumes and biosurfactants has a positive effect on soil quality by its enrichment with inorganic P and soluble phenols, while no enrichment in NO3− and NH4+ was observed.

Abstract 478: Targeted combination nano-drug delivery system to enhance anti-cancer efficacy and reduce side effects

Abstract Background: Colorectal cancer (CRC) is a global health concern, with significant morbidity and mortality. Oxaliplatin (OXL) is a common treatment for CRC but its effectiveness is hampered by side effects, including gastrointestinal and neurotoxic effects. APX3330 exerts anti-angiogenic and neuroprotective effects. It is proposed that the combined use of OXL and APX3330-loaded nanoparticles (NPs) would improve drug delivery to cancer cells with enhanced therapeutic efficacy while reducing side effects of OXL. Methods: We developed dual-loaded NPs containing OXL and APX3330 linked to a cancer targeting monoclonal antibody (NPs+mAb). We characterized the physicochemical properties of the NPs and evaluated their cytotoxicity and drug release using CRC cell lines. In vivo, we assessed their therapeutic efficiency and side effects using an orthotopic CRC mouse model. Results: NPs exhibited an average size of 229±26 nm with a zeta potential of -24.1±2.1 mV. The drug loading and encapsulation efficiency were 3.3±0.2% and 71.9%, respectively. Morphological analysis via scanning electron microscopy confirmed NP structure. In vitro drug release studies showed a slower release profile from NPs compared to free OXL, with a cumulative release of 34% (w/w) after 48 hours. Notably, cytotoxicity evaluation on CT-26 cells revealed a significant reduction in the IC50 for NPs compared to free APX3330 and OXL. In vivo studies demonstrated a remarkable 31% reduction in tumor growth rate in animals treated with NPs+mAb compared to those receiving free forms of OXL and APX3330. Body weight for animals treated with NPs+mAb remained similar to the healthy group, indicating minimal systemic toxicity. Moreover, the length of the colon in healthy organs remained steady compared to animals treated with OXL, which experienced a reduction in colon length. Animals treated with the free form of OXL exhibited diarrhoea, while the stool water content for animals treated with NPs+mAb remained at a similar level as the healthy group, suggesting reduced gastrointestinal side effects. Conclusions: This study demonstrates the potential of NPs with improved retention and encapsulation efficiency for effective drug delivery. Our research validates our novel method for creating NPs with mAb conjugation, enhancing cytotoxicity against CRC cell lines. In conclusion, the designed NPs+mAb formulation improves OXL treatment efficacy, providing targeted delivery of anti-cancer drugs to the tumor site and reducing tumor size while minimizing side effects. Citation Format: Niloufar Rashidi, Majid Davidson, Vasso Apostolopoulos, Mark R. Kelley, Kulmira Nurgali. Targeted combination nano-drug delivery system to enhance anti-cancer efficacy and reduce side effects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 478.

Comparing physiological responses of acute and chronically stressed diploid and triploid Atlantic salmon (Salmo salar)

We compared the performance and physiological response of diploid and triploid Atlantic salmon (Salmo salar) following 21 days of either unpredictable chronic stress (UCS) or no disturbance (control). Thereafter, all four groups were challenged with a novel acute stressor consisting of crowding, netting and transfer into another tank. Growth was monitored during the 21-day period, and stress response to the novel acute stress challenge was assessed by blood chemistry parameters and pituitary gene expression. Plasma parameters were analyzed before (0) and 15, 30, 45, 60, 90, 120, 240, and 300 min after acute stress induction, whereas gene expression was analyzed at 0, 30, 60, 120, and 240 min. The results showed: a) similar growth rate and condition factor reductions from UCS in triploids and diploids, b) higher post-stress plasma Acth and cortisol levels in triploids compared with diploids, c) wider post-stress fluctuations in most of the other plasma parameters for triploids compared with diploids, particularly for glucose and lactate, d) pomca1, pomcb, crf-r1a and crf-r1b mRNA levels were more upregulated in control diploids than control triploids, and e) only minor changes in pituitary gene expression from UCS in both diploids and triploids, but with a noticeable upregulation of pomca2 and crf-r1a and crf-r1b in the triploids. Our results suggest that triploid Atlantic salmon have a higher energy cost in response to stress than diploids salmon. This can make them more sensitive to suboptimal conditions and episodes of acute and chronic stress.

The novel drug candidate S2/IAPinh improves survival in models of pancreatic and ovarian cancer

Cancer selective apoptosis remains a therapeutic challenge and off-target toxicity has limited enthusiasm for this target clinically. Sigma-2 ligands (S2) have been shown to enhance the cancer selectivity of small molecule drug candidates by improving internalization. Here, we report the synthesis of a novel drug conjugate, which was created by linking a clinically underperforming SMAC mimetic (second mitochondria-derived activator of caspases; LCL161), an inhibitor (antagonist) of inhibitor of apoptosis proteins (IAPinh) with the sigma-2 ligand SW43, resulting in the new chemical entity S2/IAPinh. Drug potency was assessed via cell viability assays across several pancreatic and ovarian cancer cell lines in comparison with the individual components (S2 and IAPinh) as well as their equimolar mixtures (S2 + IAPinh) both in vitro and in preclinical models of pancreatic and ovarian cancer. Mechanistic studies of S2/IAPinh-mediated cell death were investigated in vitro and in vivo using syngeneic and xenograft mouse models of murine pancreatic and human ovarian cancer, respectively. S2/IAPinh demonstrated markedly improved pharmacological activity in cancer cell lines and primary organoid cultures when compared to the controls. In vivo testing demonstrated a marked reduction in tumor growth rates and increased survival rates when compared to the respective control groups. The predicted mechanism of action of S2/IAPinh was confirmed through assessment of apoptosis pathways and demonstrated strong target degradation (cellular inhibitor of apoptosis proteins-1 [cIAP-1]) and activation of caspases 3 and 8. Taken together, S2/IAPinh demonstrated efficacy in models of pancreatic and ovarian cancer, two challenging malignancies in need of novel treatment concepts. Our data support an in-depth investigation into utilizing S2/IAPinh for the treatment of cancer.

A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium

Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated in four different media for differentiation and subsequently characterized histomorphologically, on mRNA and on protein level in comparison to the native epithelium of the same donor horses to identify ideal culture conditions for an in vitro model system. With increasing enterocyte differentiation, the enteroids showed a reduced growth rate as well as a predominantly spherical morphology and less budding compared to enteroids in proliferation medium. Combined or individual withdrawal of stem cell niche pathway components resulted in lower mRNA expression levels of stem cell markers and concomitant differentiation of enterocytes, goblet cells and enteroendocrine cells. For eqCE, withdrawal of Wnt alone was sufficient for the generation of differentiated enterocytes with a close resemblance to the in vivo epithelium. Combined removal of Wnt, R-spondin and Noggin and the addition of DAPT stimulated differentiation of eqJE at a similar level as the in vivo epithelium, particularly with regard to enterocytes. In summary, we successfully defined a medium composition that promotes the formation of eqJE and eqCE consisting of multiple cell types and resembling the in vivo epithelium. Our findings emphasize the importance of adapting culture conditions to the respective species and the intestinal segment. This in vitro model will be used to investigate the pathological mechanisms underlying equine gastrointestinal disorders in future studies.

Vietnam's Long-run Growth: Connecting the Dots through Climate Damage Spillovers

We propose to examine how climate damage may transform Vietnam's long-run growth rate. Because of cross-country linkages forged by bilateral trade, there are two channels through which international damage spillovers may occur. First, the dynamics of partners' growth determine future trends in Vietnam's volume of exports. Second, since the domestic impact of climate change may be heterogeneous across countries, there will be a differentiated impact on export and import market shares. Both terms play a critical role in changing trade patterns that are likely to shift Vietnam's external constraint. This demand-side view of growth based on the balance-of-payments constraint is a powerful predictor of inter-country growth differences. Our study show that the consequences of climate change could equate to a 2.5% reduction in Vietnam's growth rate over the period 2020-2060. Our decomposition exercise by effect and by partner area shows that international damage spillovers result from very different individual behaviours.