Significant Differences In Growth Research Articles

Do unions provide employment protection in times of economic crisis? A natural experiment of COVID‐19

Considering the COVID‐19 pandemic as an exogenous shock, this article attempts to determine whether unions adjusted wages and working hours to protect their members during the economic crisis that it triggered. Based on 2018–20 Korean panel data, the author finds that, during the pandemic, union members were 1.9 times more likely to keep their jobs than non‐union members. However, no significant difference is found between the wage growth in the two sectors, although union members decreased their working hours more than nonunion members. The results therefore suggest that unions provided employment protection during this period by reducing working hours, not wages.

Micro texturing and laser irradiation, two stimulus of growth and differentiation to neural like cell on the PMMA polymer

Nerve injuries and neurodegenerative disorders remain serious challenges due to the poor results of nerve cell regeneration. The most promising treatment for such injuries is treatments based on stimulating cells to grow and differentiate stem cells in that area into nerve cells, but there are obstacles in controlling the growth and differentiation into fully functional nerve cells and research is ongoing to overcome these obstacles. In this study, polymethyl methacrylate (PMMA) and its laser modified surfaces were used for cell culture and differentiation into neuron-like cells. In order to show the differentiation of PC12 cells into neuron-like cells, the expression levels of the neuronal markers Map2 and Synapsin-1 were examined. The results showed the growth of PC12 on raw PMMA was increased significantly compared to the control group and micrometric parallel grooves lead to a significant difference in cell growth compared to raw PMMA. Also, the simultaneous irradiation of laser to the cell cultured on PMMA caused more cell adhesion on the surface, faster growth and differentiation of PC12 into neuron-like cells.

Determining the Growing Potential of Jatropha curcas and Pongamia pinnata under Various Water Regimes

Pakistan is the among the most populated countries of the world with predominantly arid climate facing different challenges like food insecurity, energy crises, poor health system, poverty, small forest area cover, and significant effects of global warming and climate change. However, there is still a large area with scanty vegetation. Research on devising cultivation techniques for growing more economically profitable plants in these areas will not only improve the ecological environment but also the people’s living standards. Jatropha curcas (JC) and Pongamia pinnata (PP) are two economically important species (bioenergy plants) with potential to grow on dry tropical arid land. Therefore, this study was carried out to check the hypothesis “JC and PP are the suitable species to grow in the dry tropical arid land of Pakistan due to their drought tolerance ability”. The experiment was carried out at the nursery of Bahauddin Zakariya University, Multan. One year old seedlings of both plants were used in the experiment. The design involves two plants of the same species in each row, four treatments, and three replications. The levels of drought were T1 (100% water), T2 (75% water), T3 (50% water), and T4 (25% water). A generalized linear model was applied to check the variation among species and within the treatments of the same species. There is a significant difference (p ≥ 0.05) in growth among species and within the treatments of the same species. JC showed better growth as compared to PP in water stress conditions but PP grow as well. The growth parameters of JC showed a positive response in T2 (75% water) and T3 (50% water) drought levels while the growth parameters of PP showed a positive response in treatments T1 (100% water) and T2 (75% water)as compared to other drought levels. In the case of the fresh and dry weight of JC and PP, significant difference (p ≥ 0.05) was also found among species and within the treatments. Both JC and PP are recommended to grow in the dry tropical regions of Pakistan but JC is declared as the prime specie as compared to PP.

The enhancement of photosynthetic performance, water use efficiency and potato yield under elevated CO2 is cultivar dependent.

As a fourth major food crop, potato could fulfill the nutritional demand of the growing population. Understanding how potato plants respond to predicted increase in atmospheric CO2 at the physiological, biochemical and molecular level is therefore important to improve potato productivity. Thus, the main objectives of the present study are to investigate the effects of elevated CO2 on the photosynthetic performance, water use efficiency and tuber yield of various commercial potato cultivars combined with biochemical and molecular analyses. We grew five potato cultivars (AC Novachip, Atlantic, Kennebec, Russet Burbank and Shepody) at either ambient CO2 (400 μmol CO2 mol-1) or elevated (750 μmol CO2 mol-1) CO2. Compared to ambient CO2-grown counterparts, elevated CO2-grown Russet Burbank and Shepody exhibited a significant increase in tuber yield of 107% and 49% respectively, whereas AC Novachip, Atlantic and Kennebec exhibited a 16%, 6% and 44% increment respectively. These differences in CO2-enhancement of tuber yield across the cultivars were mainly associated with the differences in CO2-stimulation of rates of photosynthesis. For instance, elevated CO2 significantly stimulated the rates of gross photosynthesis for AC Novachip (30%), Russet Burbank (41%) and Shepody (28%) but had minimal effects for Atlantic and Kennebec when measured at growth light. Elevated CO2 significantly increased the total tuber number for Atlantic (40%) and Shepody (83%) but had insignificant effects for other cultivars. Average tuber size increased for AC Novachip (16%), Kennebec (30%) and Russet Burbank (80%), but decreased for Atlantic (25%) and Shepody (19%) under elevated versus ambient CO2 conditions. Although elevated CO2 minimally decreased stomatal conductance (6-22%) and transpiration rates (2-36%), instantaneous water use efficiency increased by up to 79% in all cultivars suggesting that enhanced water use efficiency was mainly associated with increased photosynthesis at elevated CO2. The effects of elevated CO2 on electron transport rates, non-photochemical quenching, excitation pressure, and leaf chlorophyll and protein content varied across the cultivars. We did not observe any significant differences in plant growth and morphology in elevated versus ambient CO2-grown plants. Taken all together, we conclude that the CO2-stimulation of photosynthetic performance, water use efficiency and tuber yield of potatoes is cultivar dependent.

Reduced growth sensitivity to water availability as potential indicator of drought-induced tree mortality risk in a Mediterranean Pinus sylvestris L. forest

IntroductionDrought-associated tree mortality has been increasing worldwide since the last decades, impacting structure and functioning of forest ecosystems, with implications for energy, carbon and water fluxes. However, the understanding of the individual vulnerability to drought-induced mortality is still limited.MethodsWe aimed to identify the factors that triggered the mortality of the widely distributed Pinus sylvestris L. in an extensive forest area in central Spain. We compared radial growth patterns in pairs of alive and recently dead individuals that co-occur in close proximity and present similar age and size, thereby isolating the effects of size and environment from the mortality process. Temporal dynamics of growth, growth synchrony, and growth sensitivity to water availability (precipitation minus potential evapotranspiration) were compared between alive and recently dead trees.Results and discussionOver the last 50 years, although we did not detect significant differences in growth between alive and dead trees, an increase in the growth synchrony and sensitivity to water availability (i.e. slope of the climatic water balance in the growth model) was observed in all trees as drought intensity increased. 20 years before mortality, dead individuals showed lower growth synchrony and growth sensitivity to water availability than alive ones, without significant differences in growth. Recorded reduction in growth synchrony and growth sensitivity to water availability in dead trees suggests a decoupling between tree growth and climate, which could increase the risk of hydraulic failure and/or carbon starvation under increasingly arid conditions. Thus, the use of reduced growth sensitivity to water availability as potential early-warning signal of tree mortality, together with reduced growth synchrony, should be further explored, particularly in pine species in seasonally dry areas.

No trade-off between growth and OsHV-1 tolerance in Pacific oysters (Crassostrea gigas) farmed on the West Coast, USA.

An important pathogen of concern for Pacific oyster growers in the USA is ostreid herpesvirus 1 (OsHV-1). Currently, oyster stocks exist that are tolerant to OsHV-1; however, it is uncertain if a trade-off exists between their tolerance to OsHV-1 and their growth in different environments. To investigate any potential trade-offs, Pacific oyster families with varying levels of OsHV-1 tolerance were grown in a bay where OsHV-1 is endemic (Tomales Bay, CA) and in a bay where OsHV-1 is absent (Willapa Bay, WA). In Tomales Bay, we found that oysters from OsHV-1 tolerant families grew faster than oysters from OsHV-1 susceptible families, while in Willapa Bay, no statistically significant difference in growth was found between oyster families with different levels of OsHV-1 tolerance observed in Tomales Bay. These findings indicate that Pacific oysters bred to be tolerant to OsHV-1 would not be expected to have a longer time-to-market regardless of the presence of OsHV-1 in the growing environment.

Shoot Growth and Virus Elimination by Antiviral Agents in Node Cultures of Three Rose Cultivars

Background and objective: This study was carried out to examine the effects of antiviral agents on shoot growth and virus elimination during node culture in rose 'Deep Purple', 'Natal Briar' and 'Pink Beauty' infected with TRSV and ArMV.Methods: Three varieties of roses confirmed to be infected with TRSV and ArMV through ImmunoStrip® Tests were node-cultured, and the medium was treated with antiviral agents ribavirin and vidarabine at 0, 10, 20, and 40 mg⋅L<sup>-1</sup> . After 60 days of culture, shoot growth and virus infection rates were evaluated.Results: In 'Deep Purple', vidarabine treatment inhibited shoot growth and survival rate more than ribavirin, and had no effect on virus elimination. Treatment of 20 mg⋅L<sup>-1</sup> ribavirin was no significant difference in shoot growth and survival rate compared to control, and TRSV and ArMV infection rates were lowered to 25% and 0%, respectively. In 'Natal Briar', treatment of 10-20 mg⋅L<sup>-1</sup> ribavirin showed no statistical difference in shoot growth compared to the control, but the infection rates of TRSV and ArMV were 100%. The shoot growth and survival rate were greatly suppressed in treatment of 10 mg⋅L<sup>-1</sup> vidarabine, but both TRSV and ArMV infection rates were 50%. In 'Pink Beauty', the higher the treatment concentration of ribavirin and vidarabine, the more the shoot growth was suppressed. Ribavirin had no effect on TRSV elimination, and 20 mg⋅L<sup>-1</sup> vidarabine treatment showed that both TRSV and ArMV infection rates were 50%.Conclusion: Depending on the type and concentration of the antiviral agent, the elimination effects of viruses were different for each variety of rose. It was most effective to treat 20 mg⋅L<sup>-1</sup> ribavirin in 'Deep Purple', 10 mg⋅L<sup>-1</sup> vidarabine in 'Natal Briar', and 20 mg⋅L<sup>-1</sup> vidarabine in 'Pink Beauty' in media for TRSV and ArMV elimination.

Effects of enhancing sweet potatoes variety diversity in increasing yield and quality of tuber

Sweet potato is a tuber crop which is grown for both its tubers and vines. The tubers are used as human food while the vines are used as livestock feed. It is a crop that is able to resist drought conditions and will take a short time to mature hence plays a big role in food security in Kenya. There are many different varieties of sweet potatoes in the world and in Kenya which differ in growth and yield characteristics. Since most of the population in Embu West Sub County is made up of small scale farmers who undertake farming as an economic activity, it is necessary for them to adopt the variety that produces the highest yield. A study was carried out to determine effects of enhancing suit potatoes variety diversity in increasing yield and quality of tuber of SPK004, Kenspot 3 and Kenspot 4.Data on specific growth and yield parameters was collected in two seasons , summarized using excel then analysed using SPSS version 23, α=0.05. The study showed significant growth and yield potential difference in all yield parameters among the three sweet potato varieties with Kenspot 4 being the highest producer of and biomass above the ground and widest tuber diameter.

Investigation on Bacterial Growth and pH in Milk after the Expiration Date.

Food waste is a serious national and global problem. Milk is one of the most frequently wasted food items. This study aims to determine how long postexpiration-pasteurized milk may still be safe to consume and what is the relationship between bacterial growth from the milk and time after expiration. The experiments were carried out by incubating milk with agarose gel. The results showed that the bacterial growth was relatively low for at least the first few days after expiration. The more the days passed after the expiration date, the more the bacteria grew from the milk. There was no significant difference in the bacteria colony numbers in the whole milk samples opened either on day 0 or day 5 of expiration. None of the fat-free milk samples collected on the later (1-10) days showed any statistically significant difference in bacterial growth compared to the samples collected on the day after expiration (day 0). The bacterial growth increased with the increasing fat content of the milk. In addition, the rate of bacterial growth from the milk correlated with the acidity of the milk that is measured by pH. No significant sensory changes could be detected in any of the milk samples immediately after opening on the day of expiration or for up to 10 days after expiration when the unopened cartons were kept refrigerated. However, within 24 hours of opening the carton, whole milk that has expired for 6 or more days and 2% milk that has expired for 8 or more days developed a sour taste and mildly pungent smell. This sensory change was accompanied by the formation of lumps and fat-water separation. Fat-free milk samples remained unchanged under the same conditions. The experimental results suggest that whole and 2% skim milk may be safe for consumption up to 5-6 days after expiration. Fat-free milk may be safe for consumption up to 10 days after expiration, possibly longer. This study devised a way to show that milk is still safe for consumption after expiration; it will help reduce food waste.

Evaluation of drought stress level in Sargent's cherry (Prunus sargentii Rehder) using photosynthesis and chlorophyll fluorescence parameters and proline content analysis.

Sargent's cherry trees (Prunus sargentiiRehder) are widely planted as an ornamental, climate change-sensing species. This study investigated changes in the soil moisture content, fresh weight, photosynthesis and chlorophyll fluorescence properties, and the chlorophyll and proline content of four-year-old P. sargentii seedlings after 30 days of drought stress. In the trees subjected to drought stress treatment, soil moisture content decreased, and the fresh weight of the aboveground part of the plant decreased. However, there was no significant difference in the root growth of the dried plants. Among the photosynthesis parameters, Pn MAX, E and gs showed a significant (p<0.001) decrease after 15 days in dry-stressed seedlings, but there was no difference between treatments in WUE until 20 days, and there was a significant (p<0.001) difference after 24 days. Chlorophyll fluorescence parameters, Fv/Fm, ΦPSII, Rfd, NPQ, and Pn MAX, also increased after 10 days in dry-stressed seedlings, but these changes did not reach statistical significance compared to the control treatment. These results may suggest that drought stress highly correlates with photosynthesis and chlorophyll fluorescence parameters. Chlorophyll content also significantly decreased in the seedlings under drought stress compared with the control treatment. The proline content decreased until the 10th day of drought stress treatment and increased after the 15th day, showing an increase of 10.9% on the 15th day and 57.1% on the 30th day, compared to the control treatment. These results suggest that photosynthesis, chlorophyll fluorescence parameters, and proline content can be used to evaluate drought stress in trees. The results of this study can contribute to the management of forests, such as the irrigation of trees when pore control ability and photosynthesis ability decrease.

Significant reduction of grain size and texture intensity in laser powder bed fusion fabricated nickel-based superalloy by increasing constitutional supercooling

Coarse columnar grains are always present in nickel-based superalloys fabricated by laser powder bed fusion (LPBF), which not only causes anisotropy in mechanical properties, but also increases the hot cracking susceptibility of the alloys. Generally, the addition of nucleating agents is a common consideration for grain refinement. However, the intrinsic ultra-high thermal gradient and cooling rate of the LPBF process result in significant differences in grain nucleation and growth compared to conventional casting. The ultra-high thermal gradient of the LPBF process severely inhibits constitutional supercooling, which significantly reduces the heterogeneous nucleation efficiency of nickel-based superalloys. In this work, we introduced Zr into Haynes 230 alloy to improve constitutional supercooling of the LPBF molten pool during solidification and investigated the individual/synergistic effects of nucleating agent (TiC particles) and solute (Zr atoms) addition on the grain refinement of Haynes 230 alloy. It was found that the columnar grain size and texture intensity of LPBF fabricated Haynes 230 alloys were dramatically decreased with Zr and TiC co-addition, which obviously improved the mechanical properties as well as weakened its anisotropy. Our work revealed that inhibition of cell epitaxial growth and promotion of cell nucleation are necessary conditions to refine the grain size of LPBF fabricated nickel-based superalloys. Solutes with high growth restriction factor (Q value), such as Zr in our work, can rapidly generate constitutional supercooling, thereby obviously improving the heterogeneous nucleation efficiency of nucleating agents in nickel-based superalloy during the LPBF process.

FRI178 The Clinicopathological Distinction And Malignant Potential In Patients With Lipid-rich and Lipid-poor Non-functioning Adrenal Adenoma

Abstract Disclosure: P. Pengkhum: None. S. Lertlam: None. P. Cheunsuchon: None. V. Suvannarerg: None. S. Sirinvaravong: None. T. Wannachalee: None. Introduction: Adrenal incidentaloma (AI) is defined as an adrenal mass detected during imaging performed for conditions unrelated to suspected adrenal diseases. The latest evidence shows that most AIs are non-functioning adrenocortical adenomas (NFA). Of these, 70% are lipid-rich adenomas (LRA) and 30% are lipid-poor adenomas (LPA). Current guidelines do not recommend further follow-up imaging in patients with an adrenal mass size &amp;lt;4 cm with clear benign features. But some non-functioning lipid-poor lesions may carry malignant potential and proper management including surgical resection should be offered. However, the natural history and optimal specific follow-up surveillance of non-functioning LPA have not been well characterized. Objectives: We aimed to evaluate the clinical characteristics, natural courses, and histopathological data in NFA, and compared the malignant potential between LRA and LPA. Methods: We retrospectively studied all patients with AI between Jan 2005 to June 2022. Only patients with NFA who underwent adrenalectomy or were followed up by imaging for at least 6 months were included. The demographic, radiographic, and follow-up data were evaluated in all patients. Available adrenal tissues were performed for pathological assessment by using the Weiss score, Ki-67, and reticulin algorithm to identify the malignant potential. Results: Of 303 patients with AIs, 146 patients (48%) with NFAs were recruited, including 79 masses in LRA group and 77 masses in LPA group. Of these, the median age was 61 years, 43% were men, and the median follow-up period was 47 months. The initial median adenoma size was 1.6 cm [1.2-2.1]. During the follow-up period, only 3 LPAs developed significant interval enlargement. However, none of these were neither treated by surgical resection nor demonstrated malignant transformation based on imaging appearance. Overall, there was no difference in tumor growth between both groups by radiological assessment. Furthermore, 27 patients of NFAs underwent adrenalectomy; 8 masses in LRA group and 19 masses in LPA group. Of these, no difference in histological assessment in both groups. However, two large LPAs which size ≥4 cm revealed a Weiss score ≥3 with the presence of reticulin algorithm indicating potential malignant behavior. In contrast to LRA. Regardless of the adrenal mass size, the histopathological results revealed no suggestive malignant features. Conclusions: There was no significant difference in characteristics and tumor growth in patients with non-functioning LRA and LPA across 4-year interval follow-up data. Significant enlargement of adenomas was observed in few small LPAs, therefore periodic follow-up imaging should be offered in small LPAs with evidence of interval enlargement. According to our histopathological data, surgical resection should be considered in the large LPA (≥4 cm) due to its malignant potential. Presentation: Friday, June 16, 2023

Formulated feeds prolong the reproductive period of a cultured echinoid

AbstractObjectiveThe production of echinoid larvae and juveniles is required in both wild‐population enhancement programs and full life‐cycle culture. Many echinoid species of commercial interest spawn only once or twice a year, however, restricting temporal access to viable broodstock. High‐energy/high‐protein formulated feeds have been used for gonad enhancement of a variety of echinoid species, but the goal of that activity has always been to produce premature gonads, which the market requires, not the production of mature gonads for breeding. The present investigation, originally established to examine gonad enhancement, took advantage of a spawning event to assess the efficacy of a formulated feed to extend the spawning period or reduce gonad recovery time after spawning in a commercially relevant echinoid, the green sea urchin Strongylocentrous droebachiensis.MethodsUrchins held in suspended sea cages were fed either a formulated, commercial, pelleted urchin diet or bull kelp Nereocystis luetkeana over 12 weeks, with survival, somatic growth, gonad quantity (wet weight, yield), and gonad quality (presence/absence of gametes) being examined over time.ResultSurvivorship after 12 weeks was high, little difference being observed between the two diets (mean ± SE: 98.2 ± 1.1% and 95.9 ± 1.9% for kelp and formulated feed, respectively). There was no significant difference in somatic growth between the two treatments, but the formulated feed led to significantly higher gonad wet weight and yield than kelp at the end of the study. When compared with kelp, the formulated feed may have led to earlier spawning and quicker gonad recovery and prevented the complete spawning of all urchins, thus extending the period of gamete availability.ConclusionFormulated feeds may have the potential to maintain reproductively active broodstock over an extended time, allowing expansion of the period in which echinoid larvae and juveniles can be produced.

The Right Time to Add A Commercial Feed for Rabbitfish (Siganus Guttatus) from 10 to 25 Days of Age

A study on the timing of the start of supplied commercial feed for rabbitfish fry (Siganus guttatus) to determine the optimal survival rate and growth in terms of length and weight at 30 days. Four trials, 3 replicates each, and 4 batches in 2020 – 2022 differed when supplied commercial feed to fish fry at 10, 15, 20, and 25 days old before provided commercial feed, all larvae fed by zooplankton and phytoplankton. The results showed that the rabbitfish fry supplemented with commercial feed at 20 days old (NT3 (20)) showed the fastest growth in length, an average of 18.53 mm at 30 days old, average daily increase of the size of 0.65 mm/day and growth rate of size of 5.95% and 36.98 to 38.01g/head of trial 3 (TN3) (20) in 4 feeding batches, and the highest survival rate of 71.85% and weight at the final feeding stage at 30 days of age. There were significant differences in weight growth for the fish fry from at last feeding, and during the fry stages, fish tends to grow in length continuously, then increase in weight later. There was a significant impact by stomach volume at 30 days of the age of NT3 was much higher than in other trials. The other three timing trails of commercial feeding did not have any differences and showed less effectiveness than the treatment of fish at 10, 20, and 30 days old in terms of growth in length, weight, and survival rate. The stomach volume of Fries was stimulated by feedstuffs and early supplied feed to fries when they could eat; trial 3 (NT20) was the biggest stomach size, with a circumference of 11,76 mm2.

Growth Analysis of Three Soybean Varieties on Ultisol

Expansion of planting areas by utilizing sub-optimal land and the use of superior varieties is one of the alternatives in increasing soybean production in Indonesia. Therefore, this study aims to analyze the growth of three soybean varieties on Ultisol land in the Experimental Garden of the Faculty of Agriculture, Andalas University. This activity was conducted from September to November of 2022. The soybean varieties used in this study were Dena 1, Devon 1, and Demas 1. This research was designed using a Randomized Group Design with 1 factor, namely the type of variety with 3 groups of replications. The observation results of each treatment were analyzed statistically. If F count is greater than F table, then continued with Tukey's Further Test at the 5% level. It was concluded that some of the observed characters of the three soybean varieties showed that there were significant differences in growth in the observation of plant height and leaf area index. While in the observation of plant growth rate and the ratio of crown: root there was no significant difference between the tested varieties.

Tumor Regression with 5-Aminolevulinic Acid (5-ALA)-Mediated Radiodynamic Therapy (RDT) Using Different Megavoltage Energies

5-aminolevulinic acid (5-ALA) accumulates in target tumor cells, where it is metabolized to a photosensitizer - protoporphyrin IX (PpIX). Cherenkov light induced by high-energy photon beams effectively activates the PpIX due to the Soret band. The activated photosensitizers lead to cellular toxicities to kill malignant tumor cells by converting surrounding tissue oxygens into singlet oxygens. Radiodynamic therapy (RDT) utilizes cellular damage caused by both radiation dose and activated photosensitizer. The emission of Cherenkov light was observed higher with higher energy irradiation in phantom and ex-vivo tissues. Therefore, using an in-vivo mouse model, this study aimed to investigate the efficacy and energy dependency of RDT combined with 5-ALA and different ranges of megavoltage photon irradiation. In order to investigate individual and synergistic effects of 5-ALA administration and radiation treatment, the tumors (n = 344) were randomized into eight groups: control (untreated), 5-ALA only, 6-, 15-, and 45-MV conventional radiation treatment (RT) only, 6-, 15-, and 45-MV RDT. 4 Gy in a single fraction was delivered to the tumors using three different energy photons for RT only and RDT groups. 5-ALA was systemically injected into 5-ALA only and RDT groups at 100 mg/kg by tail-vein 4 hours before irradiation for endogenous PpIX accumulation in the tumor. Tumor growth was measured using a 1.5 T MR scanner on the day of treatment (prior to the treatment), 3, 7, and 14 days post-treatment. Two-way repeated ANOVA with Bonferroni correction was used to compare each treatment group to determine the statistically significant difference in tumor growth. A total of 45 MV RDT resulted in the most significant decrease in tumor growth by 58.8 ± 3.4 %, 58.0 ± 3.0 %, and 55.0 ± 3.0 % compared to 5-ALA, 45 MV RT, and control group on 7 days post-treatment (P<0.001), respectively. Moreover, a synergistic effect of 45 MV RDT causes a 47.1-54.1 % additional decrease in tumor growth toward the effective treatment outcome. Compared to the different energies, 45 MV RDT resulted in a 52.2 ± 3.1 % and 19.7 ± 7.2 % decrease in tumor growth compared to 6 MV RDT and 15 MV RDT on 7 days post-treatment, respectively (P<0.001), whereas the conventional RT was not. Radiodynamic therapy using 5-ALA administration and 45MV photon beam irradiation resulted in the most significant tumor growth control. A photon energy dependency was observed in radiodynamic therapy. 45MV photon beams showed greater in activating PpIX, improving the synergistic effects of radiation dose and activated photosensitizer than the other energies. The preliminary results provide a foundation for new innovative treatment strategies that have the potential to improve cancer treatment.

Investigation of Hydrogen Peroxide for 45MV 5-Aminolevulinic Acid-Mediated Radiodynamic Therapy

Cytotoxicity caused by the reactive oxygen species (ROS), such as singlet oxygen species (1O2), superoxide radical (O2-), and hydroxyl radical (HO), is a mechanism for treating cancer cells in radiation therapy. 5-aminolevulinic acid (5-ALA)-mediated radiodynamic therapy (RDT) is more effective in killing tumor cells than conventional radiation therapy. ROS is produced not only by ionizing radiation but also by Cherenkov light-activated protoporphyrin IX (PpIX), which is metabolized endogenously from 5-ALA. Moreover, PpIX also catalyzes hydrogen peroxide to generate 1O2, and an enhanced catalytic yield of 1O2 was observed in X-ray irradiation in vitro. Therefore, using an in-vivo mouse model, this study aimed to investigate the effect of hydrogen peroxide as a coenzyme catalyst on a novel 45MV 5-ALA-mediated RDT. A subcutaneous C57BL/6 mouse model of KP1 cell line was used. The tumors (n = 240) were randomized into six groups, consisting of untreated, conventional radiation treatment (RT), and RDT with or without hydrogen peroxide: 1. control (untreated), 2. hydrogen peroxide, 3. 45MV RT, 4. 45MV RT + hydrogen peroxide, 5. 45MV RDT, 6. 45MV RDT + hydrogen peroxide. For 45MV photon irradiation, a single fraction of 4 Gy was delivered to the tumors. 5-ALA was systemically injected at 100 mg/kg by tail-vein 4 hours before the treatment for endogenous PpIX accumulation in the tumor. Carbamide peroxide was used to deliver hydrogen peroxide to tissue and was administered at 60 mg/kg intratumorally into tumors ∼3-5 min before the treatment. The treatment effect of a single fraction of treatment was measured by calculating tumor growth, measured using a 1.5 T MR scanner on the day of treatment (prior to the treatment), 3 and 7 days post-treatment. Two-way repeated ANOVA with Bonferroni correction was used to compare each treatment group to determine the statistically significant difference in tumor growth. A total of 45MV RDT with hydrogen peroxide was shown to significantly delay the tumor growth for the mouse model and cell line investigated in this work. 45MV RDT with hydrogen peroxide group resulted in a decrease in tumor growth by 51.3 ± 4.1 % and 56.1 ± 5.1 % compared to the control group on 3 and 7 days post-treatment, respectively (P<0.001), and 43.4 ± 0.8 % and 50.9 ± 0.8 % compared to 45MV RT alone on 3 and 7 days post-treatment, respectively. Moreover, the enhancement effect of hydrogen peroxide on 45MV RDT was 2.2-4.7 times greater on 45MV RT alone (P<0.05). Hydrogen peroxide did not contribute to tumor growth when administered alone. A total of 45MV 5-ALA-mediated RDT with hydrogen peroxide resulted in the most significant tumor growth delay compared to the other groups. The catalytic effect of PpIX and hydrogen peroxide was observed in-vivo. These preliminary results demonstrate an effective cancer treatment modality.

Combination of BET Inhibition with Radiation Induces Anti-Tumor Immune Response in Pre-Clinical Models of ER-Positive Breast Cancer

Despite advances in multidisciplinary care, the risk of late recurrence in Estrogen Receptor-positive (ER+) breast cancer (BC) patients following standard-of-care treatment ranges from 20-52% at 20 years. While immune checkpoint inhibitors (CPI), have shown enormous potential in inducing long term remission in some cases, patients with ER+ BC show poor response to these agents. The objective of this study is to develop effective radiation therapy (RT)-based combination regimens that induce anti-tumor immunity against immunologically cold ER+ breast tumors. We have previously shown that pharmacological inhibition of bromodomain and extraterminal domain (BET) family of epigenetic regulator proteins overcomes both endocrine therapy resistance and radiation resistance in pre-clinical models of ER+ BC by reversing pathological reprogramming of transcriptional and DNA repair pathways, respectively. In this study, we explore the potential of RT+BET inhibition combination to elicit anti-tumor immunity against ER+ BC. A unique ER+ BC model, MXT+, was used to generate tumors in female, BDF1 mice. PD-L1 expression following RT or RT+OTX015 (a BET inhibitor) was assessed by Western blot analysis. Comet assay was used to assess the degree of unrepaired DNA damage following treatment with RT alone, OTX015 alone and RT+OTX015. Multiplex IHC was used to assess changes in tumor microenvironment after various treatments. Tumor growth characteristics were established following treatment with vehicle, RT alone (8 Gy x 3), OTX015 alone (100 mg/kg x 3 days) and RT+OTX015. Using murine ER+ BC model, MXT+, we show that RT is a potent inducer of PD-L1 expression. Combination of RT+OTX015 completely ablated RT-mediated PD-L1 induction at the transcription level. OTX015 also accentuated RT-induced DNA damage through inhibition of NHEJ pathway, resulting in increased micronuclei formation (which activate cGAS/STING pathway, a critical bridge between innate and adaptive immunity). Combination of OTX015 with RT also caused a statistically significant (on unpaired t test) increase tumor infiltrating lymphocytes. In in vivo therapeutic studies, combination of OTX015 with RT (8 Gy x 3) resulted in extraordinary synergy and caused complete inhibition of long-term tumor growth. Treatment with RT alone (8 Gy x 3) or OTX015 alone (on 3 consecutive days) only caused modest inhibition of tumor growth. ANOVA with Dunnett's test was used to adjust for multiple comparisons and showed statistically significant difference in tumor growth between OTX015+RT arm and OTX alone as well as OTX015+RT arm and RT alone. Patients with ER+ BC face high risk of late recurrence after definitive treatment and show poor response to immune CPIs. We show that combination of BET inhibition with RT induces anti-tumor immunity and completely prevents tumor growth. Clinical validation of this regimen is warranted to prevent late recurrences in ER+ BC patients.

Embryonic cranial cartilage defects in the Fgfr3Y367C /+ mouse model of achondroplasia.

Achondroplasia, the most common chondrodysplasia in humans, is caused by one of two gain of function mutations localized in the transmembrane domain of fibroblast growth factor receptor 3 (FGFR3) leading to constitutive activation of FGFR3 and subsequent growth plate cartilage and bone defects. Phenotypic features of achondroplasia include macrocephaly with frontal bossing, midface hypoplasia, disproportionate shortening of the extremities, brachydactyly with trident configuration of the hand, and bowed legs. The condition is defined primarily on postnatal effects on bone and cartilage, and embryonic development of tissues in affected individuals is not well studied. Using the Fgfr3Y367C/+ mouse model of achondroplasia, we investigated the developing chondrocranium and Meckel's cartilage (MC) at embryonic days (E)14.5 and E16.5. Sparse hand annotations of chondrocranial and MC cartilages visualized in phosphotungstic acid enhanced three-dimensional (3D) micro-computed tomography (microCT) images were used to train our automatic deep learning-based 3D segmentation model and produce 3D isosurfaces of the chondrocranium and MC. Using 3D coordinates of landmarks measured on the 3D isosurfaces, we quantified differences in the chondrocranium and MC of Fgfr3Y367C/+ mice relative to those of their unaffected littermates. Statistically significant differences in morphology and growth of the chondrocranium and MC were found, indicating direct effects of this Fgfr3 mutation on embryonic cranial and pharyngeal cartilages, which in turn can secondarily affect cranial dermal bone development. Our results support the suggestion that early therapeutic intervention during cartilage formation may lessen the effects of this condition.

Giant Duckweed (Spirodela polyrhiza) Root Growth as a Simple and Sensitive Indicator of Copper and Chromium Contamination.

Aquatic environment are often contaminated with heavy metals from various industrial sources. However, physicochemical techniques for pollutant detection are limited, thus prompting the need for additional bioassays. We investigated the use of greater duckweed (Spirodela polyrhiza) as a bioindicator of metal pollution. We exposed S. polyrhiza to four pollutants (namely, silver, cadmium, copper, and chromium) and assessed metal toxicity by measuring its frond area and the length of its regrown roots. The plant displayed significant differences in both frond size and root growth in response to the four metals. Silver was the most toxic (EC50 = 23 µg L-1) while copper the least (EC50 = 365-607 µg L-1). Direct comparisons of metal sensitivity and the reliability of the two endpoint assays showed that root growth was more sensitive (lower in terms of 50% effective concentration) to chromium, cadmium, and copper, and was more reliable (lower in terms of coefficient of variation) than those for frond area. Compared to conventional Lemna-based tests, the S. polyrhiza test is easier to perform (requiring only one 24-well plate, 3 mL of medium and a 72-h exposure). Moreover, it does not require livestock cultivation/maintenance, making it more suitable for repeated measurements. Measurements of S. polyrhiza root length may be suitable for assessment when copper and chromium in municipal and industrial wastewater exceed the environmentally permissible levels.