bluuebunny/crossref_metadata_2025 huggingface dataset:
The expression of novel PII-like protein from Lentilactobacillus hilgardii
Authors: Iskhakova Z , Zhuravleva D , Matigorova V , Kayumov A
Published: February 2022

The understanding of molecular mechanisms of bacterial adaptation to environmental changes represents the basis for maintaining the homeostasis of microbiomes of natural and artificial biotopes. Lactic acid bacteria are presented everywhere in the environment and present the indigenous microflora of various fermented substrates, fruits, soil and feces. In natural habitats the starvation by available nitrogen, carbon or energy sources can occur. Nevertheless, the regulation of their nitrogen metabolism remains unexplored and is presented only in a few works. Generally, in the most bacteria the nitrogen metabolism is regulated by PII like proteins in response to the nutrition and energy availability via sensing changes of intracellular level of ATP, ADP, 2-OG and glutamine. Among more than 400 species of Lactobacillaceae known to the date, only seven contain genes encoding for PII homologue. In Lentilactobacilli, the gene of PII-like proteins has unique genetic background being located within the potABCDN operon, which encodes the ABC transporter of spermidine/putrescine, the products of amino acids metabolism. These molecules contain multiple amino groups and thus could also serve as nitrogen source. The maximum expression of the potABCDN operon was observed upon transfer to a medium containing spermidine as solely nitrogen source, while is suppressed upon growth with glutamate and glutamine. This suggests the availability of energy supply as the leading factor determining the level of potABCDN operon transcription in L. hilgardii cells.
Resistance of several Banten local rice varieties (Oryza sativa L) towards brown planthopper (Nilaparvata lugens Stal.) attack as potential genetic resources
Authors: Hastuti D , Yenny R F , Susilawati P N , Welasih A P
Published: February 2022

The purpose of this study was to investigate the genetic resistance of 12 local varieties of rice (Oryza sativa L.) from Lebak, Banten Province, Indonesia to Brown Planthopper (BPH) (Nilaparvata lugens Stal.). The DNA amplification of PCR with SSR marker was analyzed by scaling the emerging tape and the allele size on the associated marker by using the gel analizer software and determined the percentage of polymorphism by calculating the total of polymorphic alleles in each primer. Datas were also analyzed in relation of plant familiarity patterns by using NTSys software to determine the right elders in the assembly of new improved varieties. The results showed that the varieties were suspected to have similar resistance if they had the same allele size with the differential varieties, the Sereh Variety in the Bph1 resistance genes, Tambleg variety in the Bph3 resistance genes, Cireh Gudang variety in the Bph2&3 resistance genes, and Caok Variety in the Bph4 resistance genes. Cireh Gudang varieties are suspected of having horizontal resistance as they are supported by the polygenic resistance genes in the Bph2&3 genes. Tambleg variety have the closest genetic similarities to the differential varieties, so it is presumed to be the best variety to be used as an elderly superior varieties of brown planthopper resistant, followed by Sereh and Cireh Gudang varieties.
The Effect of Bio-Fertilizer Applications Bacillus cereus and Lysinibacillus sp On Paprika Plants (Capsicum annuum L) On Plant Nutrient Content and Cultivation Media
Authors: Sodiq Abdul Hasyim , Setiawati Mieke Rochimi , Santosa Dwi Andreas , Widayat Dedi
Published: February 2022

This study conducted several applications of biofertilizer Bacillus cereus and Lysinibacillus sp on two doses of chemical fertilizer to find out the impact of the use of biofertilizers on paprika cultivation. The design of the research experiment used is a randomized design group of factorial patterns with two factors, the first factor is the use of biofertilizers with seven levels. The second factor is the use of chemical fertilizer doses consisting of two levels. Parameters observed in the laboratory include nutrient content N, P, K plant, and cultivation media. The results showed the nutrient content of P and K plants the highest yields were indicated by bio-fertilizer isolated 2 (P2) and biofertilizer consortium (P4) with a dose of chemical fertilizer 75% (A2). The total nutrient content of the planting media 2 best results are indicated by the treatment of biofertilizer isolated 1 (P1) and biofertilizer isolated 2 (P2) with 100% chemical fertilizer (A1). Bio-fertilizers Isolated Bacillus subtilis 1 and Bacillus subtilis 2 and Lysinibacillus sp. needs to be tested functionally as a nutrient-producing microbe as an absolute technical requirement of biofertilizer products by Ministry of Agriculture standards.
Internal factors and competency on the performance model of self-support agricultural extensional agents in Serang City
Authors: Fauzan A , Mirajiani  , Mulyaningsih A
Published: February 2022

This study was conducted to analyze the internal factors that affect the competence and performance of self-support agricultural extension agents in Serang City. The study was designed as explanatory research, which explains the variables’ position and the causal relationship between variables with a quantitative approach using Partial Least Square (PLS) analysis. Data collection was carried out from April to June 2021 with 34 respondents. The types of data collected were primary and secondary data. The data scale in this study was in the form of ratio data for internal factor variables and ordinal data for competence and performance variables. Based on the data processing results on the performance model of self-support agricultural extension agents in Serang City, it can be concluded that internal factors have a significant direct influence on competence with an influence value of 0.354. Internal factors also significantly influence the performance of self-support agricultural extension agents either directly with a value of 0.297 or indirectly through the competence variable with a value of 0.223. Competence also has a significant effect on performance, with a value of 0.631. Internal factors include training, experience, cosmopolitanism, and social status aspects. Competence must be enhanced to increase the performance of self-supporting agricultural educators. Competence of self-support agricultural teachers refers to the ability and capacity of self-support agricultural instructors to perform their duties. Enhancing the competence and performance of self-supporting agricultural extension workers can be accomplished by boosting their internal factors such as training, experience, and global exposure through training, internships, courses, and coordination meetings. Meanwhile, when recruiting independent agricultural extension workers, socioeconomic status might be considered.
Farmers’ perceptions towards local food in Pandeglang Regency
Authors: Mulyaningsih A , Astuti A , Sari R M
Published: February 2022

Consumption of corn, cassava, and sweet potatoes is closely related to people’s perceptions of local non-rice food. This study was conducted to determine farmers’ perceptions of local food corn, cassava, and sweet potatoes and to explore the obstacles faced in realizing food diversification. The study was conducted from February to April 2021 in Pandeglang Regency, Banten, which is the center of corn, cassava and sweet potato plants. The research method used a survey method to 270 respondents who were selected purposively. Data were analyzed descriptively by making tabulations. The results of the study indicate that the perception of farmers on local food seen from the perception of relative profit is classified as moderate, based on complexity is classified as very low, based on suitability and easy to observe is classified as very high. The obstacles faced by farmers in realizing local food diversification were the availability of raw materials for sweet potato, cassava and corn which is still very volatile because it depends on the cropping pattern and harvest season, cannot be planted continuously in one year because it is vulnerable to pests, the price fluctuating, there is no uniformity of type or variety and quality, and requires storage space for the harvest.
Evaluation on some nitrogen degrading bacteria for culture of glass ell
Authors: Widyastuti Y R , Saputra A , Yosmaniar  , Prihadi T H , Sunarno M T D
Published: February 2022

Nitrogen (N) degrading bacterias are needed for the control of fish culture organic waste. The purpose of the study was to evaluate N- degrading bacterias on survival, growth performance, blood profil, and media water quality for glass eel culture. Twelve aquaria were used at rate of 20 fish/L water. Artemia sp and peleted diet were fed at satiation on test fish for 60 days of experimental period. Completely randomized design with 4 treatments and 3 replications were used. The treatments were different N- degrading bacterias, namely: A) no bacteria (control); B) Nitrobacter sp, C) Pseudomonas sp, and D) Bacillus sp. The parameters tested were survival rate, absolute growth and length, blood profil, and water quality. The results showed that Bacillus sp was the best bacteria for degrading nitrogen to produce the best values of survival rate (90.62±0.40%), total length (9.31±0.59 cm), body weight (1.04±0.17 g/fish), total erythrocytes (1.04×106–1.10×106 cells/mm³) and total leukocytes (7.61×106–8.85×106 cells/mm³). Nitrite, nitrate, ammonia concentrations in culture added bacterias were relatively stable and lower than those at control. It could be stated that adding Bacillus sp can improve survival, growth performance, and blood profil as well as uphold water quality in optimal range conditions.
Thermal and pH Stability of the Red Seaweed (Rhodophyceae) Phycoerythrin Pigments from Kupang, Indonesia
Authors: Munifah Ifah , Musfiroh Siti , Munandar Aris , Surilayani Dini
Published: February 2022

This is a study of a red seaweed sample taken from the seawaters of Kupang, East Nusa Tenggara, Indonesia. Red seaweed utilization as a producer of bioactive compounds is potential to be developed further due to the presence of phycoerythrin (PE) pigment. The phycoerythrin (PE) pigment is extracted using phosphate buffer solvent. The objective of this study is to determine the effect of heat treatment at temperatures ranging from 30° to 80°C on the stability of the antioxidants and proteins in PE pigment. Moreover, the study will also endeavor to learn the effect of adding buffer solutions on the stability of the antioxidants and proteins in PE pigment with pH values ranging from 3 to 8. The study on the stability of K2 red seaweed phycoerythrin (PE) pigment sample from Kupang, East Nusa Tenggara revealed that it remains stable even after being exposed to a 60°C temperature and a fairly wide range of pH buffers. The K2 pigment sample has an R-PE of 0.031 ml/mL. Meanwhile, the R-PE pigment purity index is 0.041 and the yield is 0.042%.
Growth and Feed Digestibility of Sangkuriang Catfish with Corn Gluten Meal-Based Feeding
Authors: Mustahal  , Hermawan D , Syamsunarno M B , Agung L A , Nurhaliza A S , Rachmawati R F , Anggraeni D , Putra A N
Published: February 2022

Diet testing was conducted for 50 days to investigate the growth and feed digestibility of Sangkuriang catfish with corn gluten meal (CGM)-based feeding. This involved 5 treatments with different levels of CGM in the feed 0, 75, 150, 225, and 300 g.kg−1 for three replications. The result showed that the value of SGR ranged from 0.87 to 1.51 %.day−1, the SGR values with CGM at 0, 75 and 150 g.kg−1 were significantly higher than the values in CGM 225 and 300 g.kg−1. The best value of protein and lipid digestibility were recorded with CGM at 0, 75 and 150 g.kg−1 compared to CGM at 225 and 300 g/kg. A greater propensity for higher CGM levels to negatively impact on growth and feed digestibility of Sangkuriang catfish. Therefore, CGM is confirmed to process the potential for application at a level of 150 g.kg−1 in the Sakuriang catfish diet without negatively influencing growth and feed digestibility.
Effectiveness of Different Mesh Size To Release Small Blue Swimming Crab on Trap Fishing
Authors: Nurdin H S , Susanto A , Saputra D Y A
Published: February 2022

Blue swimming crab (Portunus pelagicus) is a fishery commodity with important economic value and also become the main commodity for export needs. However, crab production dominantly comes from fishing activities using traps and gillnet. The fisher use traps without escape gaps so that small crabs have no chance of escaping from the traps. One alternative way to improve the crab trap selectivity is the use of larger mesh sizes on the wall of the trap. This study aimed to determine the effective mesh size for escaping small crabs. This study was conducted in Fisheries Laboratory, University of Sultan Ageng Tirtayasa from July to December 2020. We used different mesh sizes on the trap wall including of 2.0; 2.5; and 3 inches. For all experiments, we used 90 crabs with an average carapace width of 72 mm. The results showed that the pattern of crab when escaping through the net at each mesh size was varied. The crab crept up the netting wall before attempting to escape. The fastest time of crab escapes was found on a 3 inch mesh size with an average time of 10 seconds. Moreover, the highest percentage of crabs that can pass was obtained at the 3-inch mesh size with the value 60%. Based on these results, to increase the catch selectivity we recommend a 3-inch mesh to be used in the collapsible trap wall for blue swimming crab fishing.
Physical properties of bioplastic agar/chitosan blend
Authors: Agusman  , Fransiska D , Murdinah  , Wahyuni T , Irianto H E , Priambudi P , Fateha  , Abdullah A H D , Nissa R D , Firdiana B , Nurhayati
Published: February 2022

Thermoplastic agar/chitosan blend was prepared by a rheomix process, and the effect of concentration agar: chitosan on the material properties forms were investigated. This research aimed to observe the physical properties of bioplastic agar/chitosan produced through rheomix. Bioplastic was produced by mixing 65 % solid material (agar: chitosan) and 35% glycerol, the ration through the melting process at an initial temperature of 100°C. The weight ratio of agar : chitosan was 65% : 0% (F0/control), 52% : 13% (F1), 39 % : 26% (F2), 26 %: 39% (F3). The tensile strength and elongation of bioplastic were decrees with a higher chitosan concentration. The color and the opacity of bioplastic were affected by the ratio of agar: chitosan. The moisture content and the water vapor transmission rate (WVTR) of bioplastic were increased by increasing the chitosan ratio. The SEM micrograph of bioplastic showed that the blend agar and chitosan seem to have a rough and inhomogeneous surface than pure bioplastic agar. The best blend was bioplastic with agar: chitosan 52%: 13% (F1), which have properties of : tensile strength 4.95 ± 0.56 MPa, elongation at break 34.94 ± 0.99 %, moisture content 15.33 ± 0.43%, WVTR 3259.39 ± 28.12 g/m2.24 h.
Retinal age gap as a predictive biomarker of future risk of Parkinson’s disease
Authors: Hu Wenyi , Wang Wei , Wang Yueye , Chen Yifan , Shang Xianwen , Liao Huan , Huang Yu , Bulloch Gabriella , Zhang Shiran , Kiburg Katerina , Zhang Xueli , Tang Shulin , Yu Honghua , Yang Xiaohong , He Mingguang , Zhu Zhuoting
Published: February 2022

Introduction

retinal age derived from fundus images using deep learning has been verified as a novel biomarker of ageing. We aim to investigate the association between retinal age gap (retinal age–chronological age) and incident Parkinson’s disease (PD).

Methods

a deep learning (DL) model trained on 19,200 fundus images of 11,052 chronic disease-free participants was used to predict retinal age. Retinal age gap was generated by the trained DL model for the remaining 35,834 participants free of PD at the baseline assessment. Cox proportional hazards regression models were utilised to investigate the association between retinal age gap and incident PD. Multivariable logistic model was applied for prediction of 5-year PD risk and area under the receiver operator characteristic curves (AUC) was used to estimate the predictive value.

Results

a total of 35,834 participants (56.7 ± 8.04 years, 55.7% female) free of PD at baseline were included in the present analysis. After adjustment of confounding factors, 1-year increase in retinal age gap was associated with a 10% increase in risk of PD (hazard ratio [HR] = 1.10, 95% confidence interval [CI]: 1.01–1.20, P = 0.023). Compared with the lowest quartile of the retinal age gap, the risk of PD was significantly increased in the third and fourth quartiles (HR = 2.66, 95% CI: 1.13–6.22, P = 0.024; HR = 4.86, 95% CI: 1.59–14.8, P = 0.005, respectively). The predictive value of retinal age and established risk factors for 5-year PD risk were comparable (AUC = 0.708 and 0.717, P = 0.821).

Conclusion

retinal age gap demonstrated a potential for identifying individuals at a high risk of developing future PD.
Seizures initiate in zones of relative hyperexcitation in a zebrafish epilepsy model
Authors: Niemeyer James E , Gadamsetty Poornima , Chun Chanwoo , Sylvester Sherika , Lucas Jacob P , Ma Hongtao , Schwartz Theodore H , Aksay Emre R F
Published: February 2022

Seizures are thought to arise from an imbalance of excitatory and inhibitory neuronal activity. While most classical studies suggest excessive excitatory neural activity plays a generative role, some recent findings challenge this view and instead argue that excessive activity in inhibitory neurons initiates seizures.

We investigated this question of imbalance in a zebrafish seizure model with two-photon imaging of excitatory and inhibitory neuronal activity throughout the brain using a nuclear-localized calcium sensor. We found that seizures consistently initiated in circumscribed zones of the midbrain before propagating to other brain regions. Excitatory neurons were both more prevalent and more likely to be recruited than inhibitory neurons in initiation as compared with propagation zones. These findings support a mechanistic picture whereby seizures initiate in a region of hyperexcitation, then propagate more broadly once inhibitory restraint in the surround is overcome.
Cortical thickness differences are associated with cellular component morphogenesis of astrocytes and excitatory neurons in nonsuicidal self-injuring youth
Authors: Cai Suping , Guo Zitong , Wang Xuwen , Huang Kexin , Yuan Kai , Huang Liyu
Published: February 2022

Nonsuicidal self-injury (NSSI) generally occurs in youth and probably progresses to suicide. An examination of cortical thickness differences (ΔCT) between NSSI individuals and controls is crucial to investigate potential neurobiological correlates. Notably, ΔCT are influenced by specific genetic factors, and a large proportion of cortical thinning is associated with the expression of genes that overlap in astrocytes and pyramidal cells. However, in NSSI youth, the mechanisms underlying the relations between the genetic and cell type-specific transcriptional signatures to ΔCT are unclear. Here, we studied the genetic association of ΔCT in NSSI youth by performing a partial least-squares regression (PLSR) analysis of gene expression data and 3D-T1 brain images of 45 NSSI youth and 75 controls. We extracted the top-10 Gene Ontology terms for the enrichment results of upregulated PLS component 1 genes related to ΔCT to conduct the cell-type classification and enrichment analysis. Enrichment of cell type-specific genes shows that cellular component morphogenesis of astrocytes and excitatory neurons accounts for the observed NSSI-specific ΔCT. We validated the main results in independent datasets to verify the robustness and specificity. We concluded that the brain ΔCT is associated with cellular component morphogenesis of astrocytes and excitatory neurons in NSSI youth.
Bone marrow activation in response to metabolic syndrome and early atherosclerosis
Authors: Devesa Ana , Lobo-González Manuel , Martínez-Milla Juan , Oliva Belén , García-Lunar Inés , Mastrangelo Annalaura , España Samuel , Sanz Javier , Mendiguren José M , Bueno Hector , Fuster Jose J , Andrés Vicente , Fernández-Ortiz Antonio , Sancho David , Fernández-Friera Leticia , Sanchez-Gonzalez Javier , Rossello Xavier , Ibanez Borja , Fuster Valentin
Published: February 2022

Aims

Experimental studies suggest that increased bone marrow (BM) activity is involved in the association between cardiovascular risk factors and inflammation in atherosclerosis. However, human data to support this association are sparse. The purpose was to study the association between cardiovascular risk factors, BM activation, and subclinical atherosclerosis.

Methods and results

Whole body vascular 18F-fluorodeoxyglucose positron emission tomography/magnetic resonance imaging (18F-FDG PET/MRI) was performed in 745 apparently healthy individuals [median age 50.5 (46.8–53.6) years, 83.8% men] from the Progression of Early Subclinical Atherosclerosis (PESA) study. Bone marrow activation (defined as BM 18F-FDG uptake above the median maximal standardized uptake value) was assessed in the lumbar vertebrae (L3–L4). Systemic inflammation was indexed from circulating biomarkers. Early atherosclerosis was evaluated by arterial metabolic activity by 18F-FDG uptake in five vascular territories. Late atherosclerosis was evaluated by fully formed plaques on MRI. Subjects with BM activation were more frequently men (87.6 vs. 80.0%, P = 0.005) and more frequently had metabolic syndrome (MetS) (22.2 vs. 6.7%, P < 0.001). Bone marrow activation was significantly associated with all MetS components. Bone marrow activation was also associated with increased haematopoiesis—characterized by significantly elevated leucocyte (mainly neutrophil and monocytes) and erythrocyte counts—and with markers of systemic inflammation including high-sensitivity C-reactive protein, ferritin, fibrinogen, P-selectin, and vascular cell adhesion molecule-1. The associations between BM activation and MetS (and its components) and increased erythropoiesis were maintained in the subgroup of participants with no systemic inflammation. Bone marrow activation was significantly associated with high arterial metabolic activity (18F-FDG uptake). The co-occurrence of BM activation and arterial 18F-FDG uptake was associated with more advanced atherosclerosis (i.e. plaque presence and burden).

Conclusion

In apparently healthy individuals, BM 18F-FDG uptake is associated with MetS and its components, even in the absence of systemic inflammation, and with elevated counts of circulating leucocytes. Bone marrow activation is associated with early atherosclerosis, characterized by high arterial metabolic activity. Bone marrow activation appears to be an early phenomenon in atherosclerosis development.

[Progression of Early Subclinical Atherosclerosis (PESA); NCT01410318].
Long-Term Trajectories of Frailty and Its Components After Kidney Transplantation
Authors: Chu Nadia M , Ruck Jessica , Chen Xiaomeng , Xue Qian-Li , Norman Silas P , Segev Dorry L , McAdams-DeMarco Mara A
Published: February 2022

Background

Frailty is common and associated with poor outcomes among kidney transplant (KT) recipients. While frailty improves in the first 3 months post-KT with restored kidney function, longer-term trajectories are likely to plateau/decline due to aging and other stressors (eg, immunosuppression). We evaluated longer-term post-KT trajectories of the physical frailty phenotype (PFP) and its components in adult patients at 2 centers.

Methods

PFP components were measured at admission, 1, 3, 6 months, 1 year, and annually thereafter post-KT. We used adjusted mixed-effects models to describe repeated measures of continuous components (weight, gait speed, grip strength, activity) and generalized estimating equations to quantify longitudinal, binomial response patterns (PFP; exhaustion).

Results

Among 1 336 recipients (mean age = 53) followed for a median of 1.9 years (interquartile range [IQR] = 0.1–3.2), likelihood of frailty declined in the first 2.5 years post-KT (adjusted odds ratio [aOR] = 0.96, 95% confidence interval [CI]: 0.95, 0.98), but increased after 2.5 years post-KT (aOR = 1.03, 95% CI: 1.00, 1.05). In the first 2.5 years post-KT, recipients demonstrated increases in weight (0.4 lbs/month, 95% CI: 0.3, 0.5), grip strength (0.2 kg/month, 95% CI: 0.1, 0.2), and activity (23.9 kcal/month, 95% CI: 17.5, 30.2); gait speed remained stable (−0.01 s/month, 95% CI: 0.01, 0.003). Additionally, likelihood of becoming exhausted declined post-KT (OR = 0.99, 95% CI: 0.98, 1.00). After 2.5 years post-KT, recipients demonstrated decreased grip strength (−0.07 kg/month, 95% CI: −0.12, −0.01) and activity (−20 kcal/month, 95% CI: −32.3, −8.2); they had stable weight (−0.003 lbs/month, 95% CI: −0.17, 0.16), gait speed (−0.003 s/month, 95% CI: −0.02, 0.01), and likelihood of becoming exhausted (OR = 1.01, 95% CI: 0.99, 1.02).

Conclusion

Despite frailty improvements in the first 2.5 years, recipients’ frailty worsened after 2.5 years post-KT. Specifically, they experienced gains in strength, activity, and exhaustion in the first 2.5 years post-KT, but declined in strength and activity after 2.5 years post-KT while experiencing persistent slowness. Clinicians should consider monitoring recipients for worsening frailty after 2.5 years despite shorter-term improvements.
The Moho reflectivity of the subduction beneath the Southwestern Alps from ambient seismic noise autocorrelations
Authors: Ashruf T N , Morelli A
Published: February 2022

The Western Alps shows a complex crustal organization due to the subduction of the European Plate beneath the Adriatic Plate and exhumation of the mantle wedge. The lithospheric structure of the Western Alps, that may hold significance for understanding orogenic processes and evolution, has been the subject of many geophysical studies, but the Moho profile remains unclear and this has led to controversies about the depth and extent of the European Plate beneath the Adriatic Plate. With the goal of retrieving detailed information on crustal constitution, we use autocorrelation of seismic ambient noise as a tool to map the body wave reflectivity structure at the subduction zone under the southwestern Alps. We use data recorded by the China–Italy–France Alps (CIFALPS) seismic transect, that includes 45 stations located approximately 5–10 km apart along a profile crossing the Alpine continental subduction in the Western Alps. We analyse the data set in four different frequency bands between 0.09 and 2 Hz. We automatically pick the arrival time of the Moho reflection in the second derivative of the envelope of the autocorrelation stack using prior Moho information. The 0.5–1 Hz frequency band mostly gives the best result due to the clear changes in reflectivity along the waveforms of the autocorrelation stacks after the picked arrival times of the Moho reflections. We find spatial coherence between 18 and 23 km depth in the western portion of the profile, indicating relatively homogeneous crustal rocks, and highly reflective structure under the central mountain range, due to the existence of a highly faulted zone. The very thin crust and the underlying mantle wedge known as the Ivrea body show instead high transparency to seismic waves and absence of reflections. The subduction profile of the European Plate shows a steep trend as compared to previous studies. We discuss autocorrelation stacks and Moho depths obtained from the arrival times of the picked reflectivity change in comparison with previous studies to validate the different reflection structures. Stacked ambient noise autocorrelations reliably image varied crustal properties and reflectivity structures in the highly heterogeneous region of the southwestern Alps.
A3 TARGETING PROLINE METABOLISM TO OVERCOME TREATMENT RESISTANCE IN ESOPHAGEAL CANCER
Authors: Douchin J , Nogueira de Almeida L , Gonneaud A , Boisvert F , Dufour A , Giroux V
Published: February 2022

Background

Patients with esophageal malignancy have a 5-year survival rate of only 14% in Canada. This high mortality rate is due to three factors: late diagnosis, difficulty to surgically remove the tumor due to its localization and treatment resistance. Treatment resistance has been ascribed to the presence of cancer stem cells (CSCs) inside the tumor. However, no treatment specifically directed against CSCs is available to patients. Therefore, targeting CSCs is a promising strategy to improve survival of patients with esophageal squamous cell carcinoma (ESCC), the most common type of esophageal cancer worldwide.

Aims

Herein, we developed an unbiased approach to identify new players in chemotherapy and radiotherapy resistance in ESCC.

Methods

We established radioresistant (RR), chemoresistant (CR) and radiochemoresistant (RCR) human ESCC cell lines using weekly radiation and/or continuous treatment with increasing doses of chemotherapeutic agent 5-FU. We validated that the process of resistance acquisition correlates with enrichment in CSCs as revealed by higher ALDH1 expression, and increased proportion of ALDH1high cells and CD24high/CD44high cells in flow cytometry. We then used a proteomic approach to identify new players in treatment resistance.

Results

Interestingly, pathway analysis demonstrated enrichment in energy metabolism as well as amino acid metabolism. Seahorse assays showed a more quiescent metabolism in all three types of resistant cells compared to the control cell line. More precisely, resistant cell lines have a lower respiration rate than control cell line, while glycolysis remains unchanged. Surprisingly, our results show a metabolic rewiring very different from the well-known Warburg effect. To further characterise these metabolic changes, we performed an unbiased metabolomic pilot study and confirmed a decrease in amino acid levels such as proline, in resistant cell lines. Preliminary data show that when cultured in DMEM with proline addition, CD44high/CD24high cell proportion is decreased in control and RR cell lines suggesting that proline is a key regulator of CSC population in ESCC.

Conclusions

To conclude, our results suggest an important role of metabolism in ESCC treatment resistance. This study is a first step towards the identification of new targets to fight treatment resistance in ESCC patients.

Funding Agencies

CAG, CIHRCanada research chair TIER 2
A7 FOXL1+ TELOCYTES IN MOUSE COLON ORCHESTRATE ECM BIODYNAMICS AND WOUND REPAIR RESOLUTION
Authors: Pomerleau V , Reyes-Nicolas V , Jurkovic C , Boisvert F , Perreault N
Published: February 2022

Background

The extracellular matrix (ECM) is a complex assembly of proteins that provide mechanical and biochemical stimuli to the epithelial and mesenchymal cells of the GI mucosa. Deficiencies in ECM assembly, protein production or excessive accumulation can lead to multiples pathologies including fibrosis and cancer. FoxL1+-Telocytes (TCFoxL1+) are subepithelial cells that form a network underneath the epithelium, contributing to the microenvironment that supports epithelial and immune cell homeostasis. We have previously shown that BMPR1A signaling deletion in TCFoxL1+ influences the microenvironment via stromagenesis, immune infiltration and colonic dysplasia in mouse model of GI diseases. However, the precise molecular and mechanical events that contributes to the onset of this state have yet to be elucidated.

Aims

Characterize the modulations in ECM biodynamics induced by BmpR1a-deficient TC FoxL1+ ( BmpR1a△FoxL1+) in mouse colon submucosa.

Methods

Matrisomics was performed to determine the inventory of ECM proteins expressed solely in the GI stromal compartment following tissue deconstruction of control and BmpR1a△FoxL1+ mice colons. Histological and biochemical methods were used to further characterize the collagen network and matrisome-associated modulations. Fluorescence In Situ Hybridization (FISH) was performed to study the bacterial presence in the mucosa.

Results

The set of identified proteins shows an enrichment for proteins involved in collagen network regulation, wound repair homeostasis and immune regulation such as Col1a2, Col3a1, Col6a4 and Coll14a1, as well as SerpinH1, MFAP4, ANXA1 and S100A9. Collagen network is affected with increased deposition and reorganization of fiber alignment. Unfolded collagen content was also increased in dysplastic areas of BmpR1a△FoxL1+ mouse colon with a concomitant increase in the collagen-chaperone SerpinH1. Validations of other targets indicate that BmpR1a△FoxL1+ mice deals with some type of tissue micro-injury and inflammation that is unresolved, creating a unfavorable microenvironment for tissue homeostasis.

Conclusions

Taken together, these results suggest that Bmp-signaling deficient TCFoxL1+ significantly contribute to the collagen network biodynamics through increased collagen deposition, fiber alignment reorganization and regulation of the collagen triple-helix assembly. Other matrisome modulations suggest a state of unresolved wound healing due to tissue injury, that could be the etiology of GI pathology and lead to more severe conditions upon various environmental triggers.

Funding Agencies

CIHR
A11 ISOLATION OF NON-POLAR METABOLITES IN EXCRETORY/SECRETORY PRODUCTS FROM PARASITIC HELMINTHS AND THEIR POTENTIAL AS IMMUNOTHERAPY IN INFLAMMATORY BOWEL DISEASE
Authors: Siciliani E A , Leroux L , Tam M , Arai T , Urban J F , Martin R J , Geary T G , Stevenson M , Lopes F , Jardim A
Published: February 2022

Background

Parasitic helminths are known to modulate host immune responses. This is thought to be mediated by their secretome. We are interested in the excretory/secretory products and mechanisms for modulating immune dysfunction in autoinflammatory diseases.

Aims

This research studies the mechanisms of immune modulation by parasitic helminths in the context of IBD. We aim to describe immunomodulatory helminth-derived metabolites (ESM).

Methods

Helminth-conditioned media was used to isolate ESM, which were further purified using column chromatography.

Bone marrow (BM) derived macrophages (BMDM) from C57BL6 mice, were treated with ESP fractions from Trichuris suis, Ascaris suum, Heligmosomoides polygyrus bakeri or Dirofilaria immitis, stimulated with LPS, and secreted cytokine levels measured. Moreover, BM was cultured with or without ESM throughout differentiation to BMDM.

Colitic mice (3% DSS, 5 days) were treated with A. suum ESM or PBS once daily IP. Colon lengths and TNFα mRNA were measured, and histological preparations were scored to assess pathology.

Bioactive D. immitis ESM were fractionated using preparatory HPLC and assayed for bioactivity. Active fractions were analysed using MS/MS and fragmentation patterns and molecular weights were obtained. The active fractions are currently being studied by NMR to deduce a structure of an active metabolite.

Results

BMDM treated with crude ESM decreased TNFα secretion and increased IL-10. BMDM precursors which were treated with A. suum ESM throughout differentiation had reduced proliferation in a dose dependent manner. These BMDM showed remodeling of BMDM metabolic pathways. Intracellular ROS production was inversely proportional to Alamar blue oxidation.

We found that ESM from A. suum improved DSS-colitis. Specifically, mice with DSS-induced colitis given IP ESM had longer colons, lower histolopathology score, and lower TNFα mRNA expression in gut tissue.

HPLC-fractionated D. immitis ESM used to treat BMDM yielded varying suppression of TNFα with LPS stimulation. MS/MS of TNFα suppressive fractions contained masses with fragmentation patterns which were detected in fractions of several of the above-mentioned parasite species. Preliminary NMR studies will determine if this represents a conserved structure.

Conclusions

Helminth-derived components can immunologically polarize a response in vitro, as well as favour recovery in DSS colitis. Through multiple purification steps, a nearly pure fraction is found to have bioactivity, suggesting a single, bioactive molecule that is conserved across several parasitic helminths. These data are important in understanding the host-parasite interaction modulated by ESM, as well as provide therapeutic potential in IBD.

Funding Agencies

NSERC, FRQNT
A14 THE INTESTINAL MICROBIOTA CONTRIBUTES TO AGE-RELATED MEMORY DECLINE
Authors: Kraimi N , De Palma G , Lu J , Bowdish D , Verdu E , Sibille E , Prevot T , Collins S M , Bercik P
Published: February 2022

Background

Age-related deterioration of cognitive function and memory capacity occur in both humans and rodents. For example, significant memory deficits have been reported in conventionally raised (SPF) old mice compared to conventionally raised young mice submitted to a spatial memory task (Prevot et al., 2019, Mol Neuropsychiatry 5, 84–97). Microbiota-to-brain signaling is now well established in mice and humans, but the extent to which it influences age-associated memory decline is unknown.

Aims

Our study examines whether the intestinal microbiota contributes to age-associated changes in brain function. We address the specific hypothesis that age-associated cognitive decline is attenuated in the absence of the intestinal microbiota.

Methods

We assessed anxiety-like and depressive-like behavior, locomotor activity and spatial memory performance in young germ-free (GF) mice (2–3 months of age, n=24) and senescent GF mice (13–27 months old, n=22) maintained in axenic conditions, and compared them to conventionally raised (SPF) mice of the same age. Anxiety-like behavior, locomotor activity and depressive-like behavior were measured using the light-dark preference, open-field, and tail suspension tests. We also used the Y-maze test based on a spontaneous alternation task to assess cognition, with the alternation rate as a proxy of spatial working memory. The age-associated inflammation was assessed with IL-6 cytokine plasma concentrations measured by ELISA.

Results

Anxiety-like behavior and depressive-like behavior did not change with the age regardless of the microbial status. However, old SPF mice traveled less distance (866.8 cm) than young SPF mice (1375 cm, p < 0.01) in the open-field. Similarly, old GF mice also traveled less distance (458.9 cm) than young GF mice (875.7 cm, p < 0.0001). In contrast to old SPF mice, old GF mice did not show memory impairment in the spatial memory task. Indeed, old SPF mice displayed lower alternation rate of 58.3%, compared to that found in young SPF mice (76.9%, p < 0.05) while both old and young GF mice had an identical alternation rate of 73.3% ( p > 0.05). In addition, IL-6 plasma levels revealed that old GF mice did not show signs of age-associated inflammation that was evident in old SPF mice (3.68 vs. 13.93 pg/ml, p < 0.05).

Conclusions

We conclude that the absence of age-related memory deficit in old germ-free mice is consistent with a role for the microbiota in age-related cognitive decline, likely mediated via the immune system, as suggested by the absence of age-associated inflammation in germ-free mice. We propose that novel microbiota-targeted therapeutic strategies may prevent or delay the cognitive decline of aging.

Funding Agencies

CIHRBalsam Family Foundation
A17 TRP53 REGULATES ESLF-RENEWAL AND CELL DIFFERENTIATION IN KRT15+ INTESTINAL STEM CELLS
Authors: Dubey A , Gonneaud A , Giroux V
Published: February 2022

Background

Different pools of stem cells assure the intestinal epithelium homeostasis and regeneration. We recently reported that Krt15+ intestinal cells harbor self-renewal, multipotent and regenerative capacities, characteristics consistent with a stem cell population. p53 has been associated with the development of various types of cancer, and recent studies suggested it also regulates adult stem cell behavior in some tissues. However, its role in the maintenance of intestinal stem cells has not been explicitly covered. Therefore, we hypothesize that Trp53 loss specifically in Krt15+ intestinal stem cells will perturb the epithelial homeostasis and differentiation.

Aims

Identify the role of Trp53 in the regulation of intestinal stem cell and intestinal homeostasis

Methods

To induce Trp53 loss specifically in Krt15+ cells, we generated Krt15 CrePR1;Trp53fl/fl ( Krt15△Trp53) mice and induced Cre recombination by injecting RU486 (PR agonist). Mice were euthanized after Cre recombination at different time points. Organoid cultures were established from control and experimental mice.

Results

Two-month post Cre recombination, we observed no major morphological changes in the intestinal epithelium. However, we observed that crypts isolated from Krt15△Trp53 mice died rapidly and had difficulties forming organoids. When hyperactivating the Wnt/beta-catenin pathway through CHIR treatment, we observed that budding capacity was maintained in experimental organoids while control organoids formed mainly cysts as expected. We also observed an increased in Paneth cell-specific genes, suggesting an enrichment in Paneth cells. Interestingly, the expression of EpHB2 receptor and its target genes was decreased in organoids derived from Krt15△Trp53 mice. Ephrin signaling plays a crucial role in intestinal homeostasis by regulating cell position and migration, and previous studies suggest that ephrin receptors are a target of p53. Indeed, twelve-month post Cre recombination, altered architecture of crypt and villi was noted in the small intestinal epithelium of Krt15△Trp53 mice. Differentiation towards secretory cell types, specifically Paneth, goblet, and tuft cells, are significantly increased in Krt15△Trp53 mice. Interestingly, we also observed crypt cells expressing both goblet and Paneth cell markers suggesting dysfunction in the cell fate decision. Finally, decreased Notch pathway activation was also observed supporting that Trp53 loss, specifically in Krt15+ cells, leads to a dysregulation of secretory cell fate.

Conclusions

Loss of Trp53 specifically in Krt15+ cells affects the small intestinal homeostasis and secretory cell differentiation. Decreased Notch and ephrin signaling support the possible role of Trp53 in cell fate decision and self-renewal.

Funding Agencies

None
A18 DEVELOPMENT OF CHRONIC ILEITIS IN THE TNFΔARE MOUSE IS ASSOCIATED WITH A LOSS OF MESENTERIC LYVE1+ MACROPHAGES
Authors: Keane K P , Almutairi K , Roizes S , Stephens M , von der Weid P
Published: February 2022

Background

Crohn’s disease (CD) is a form of inflammatory bowel disease that causes transmural inflammation of any part of the gastrointestinal (GI) tract but preferentially the terminal ileum and/or the colon. This chronic inflammation is not limited to the GI tract but also affects the mesentery of inflamed regions where it induces the formation of tertiary lymphoid organs (TLOs). The homeostatic LYVE1+ macrophages (LYmΦ) have been identified in many different organs including the mesentery. They have been shown to typically align along blood vessels where they participate in the control of collagen deposition and during inflammation, regulate vascular permeability and immune cell recruitment. Here we investigate the role of the mesenteric LYmΦ in a transgenic model of CD, the TNFΔARE mouse which develops terminal ileitis over time and expresses similar mesenteric alterations (TLOs) as seen in CD.

Aims

To determine whether the mesenteric LYmΦ population is altered during the development of chronic inflammation in the TNFΔARE mouse and whether they participate in the development of TLOs.

Methods

Confocal immunofluorescent imaging of the terminal ileal mesentery of TNFΔARE mice and littermate controls were preformed to identify changes to the populations of LYmΦ during the development of chronic inflammation. Myeloperoxidase activity was used to assess terminal ileitis.

Results

During the initial stage of ileitis (8 weeks) LYmΦs are present across the terminal ileal mesentery in both TNFΔARE and age-matched WT littermate controls with a subpopulation aligning along the collecting lymphatic vessels of the mesentery (n=6). At 20 weeks, while the ileitis progresses, aggregates of CD45+ cells have formed within the TNFΔARE mice where LYmΦs also accumulate (n=5). At 28 weeks when the ileitis worsens, the LYmΦs are greatly reduced from the terminal ileal mesentery as well as from the CD45+ cell aggregates (n=5). WT littermate controls never develop large CD45+ cell aggregates (n=5 for each time point). Antibodies to the LYVE1 cytosolic and extracellular domains confirmed the loss of the LYVE1 receptor over receptor cleavage, however, there was no correlation of altered collagen-1 deposition within the mesentery at 28 weeks.

Conclusions

Here we show a spatial relationship between LYmΦs and collecting lymphatic vessels as well as a decrease in LYmΦs during the development of chronic ileitis in the TNFΔARE mouse. Whether they contribute to TLO formation, or their loss exacerbates terminal ileitis is still unknown and under current investigation.

Funding Agencies

CCC
A23 ESTIMATING INDIRECT AND OUT-OF-POCKET COSTS IN PEDIATRIC INFLAMMATORY BOWEL DISEASE: A NATION-WIDE CROSS-SECTIONAL ANALYSIS
Authors: El-Matary W , Witt J , Bernstein C N , Jacobson K , Mack D R , Otley A , Walters T , Huynh H Q , deBruyn J , Griffiths A , Benchimol E I
Published: February 2022

Background

Identifying disease-related costs is a crucial step to plan for proper allocation of resources and future healthcare services for persons with inflammatory bowel disease (IBD). Data on pediatric inflammatory bowel disease-associated costs are limited.

Aims

We aimed to estimate indirect and out of pocket (OOP) pediatric IBD-associated costs in Canada.

Methods

In a nation-wide cross-sectional analysis, caregivers of children and young adults (<17 years) with IBD were invited to complete a questionnaire on lost work hours and OOP costs related to IBD in the 4 weeks prior to the survey. Participants were re-invited to periodically answer the same questionnaire every 3–9 months for 2 years. Lost productivity was calculated using the Human Capital method. Costs were reported in 2018 inflation-adjusted Canadian dollars. Predictors of high cost users (top 25%) were examined using negative binomial regression.

Results

Consecutive 243 (82 incident cases) of 262 (92.7%) approached participants completed the first survey with a total of 450 surveys longitudinally completed over 2 years. The annual median indirect costs per patient were $5,951 (IQR $1,812- $12,278), with $5,776 (IQR $1,465-$11,733) for Crohn’s disease (CD) and $6,084 (IQR $2,470-$13,371) for ulcerative colitis (UC) (p=0.77). The annual median per patient OOP costs were $2,925 (IQR $978- $8,125) with $3,021 (IQR $978- $8,125) for CD and $2,600 (IQR $975- $8,125) for UC (p=0.55). Older age (10-17y) at diagnosis (p=0.04) and parents in part-time employment (p=0.01) were predictors of higher indirect costs, while female sex (p<0.001), parents with a lower education level (p<0.001) and lower annual family income (p<0.01) were associated with higher OOP costs.

Conclusions

Indirect and OOP IBD-associated costs are substantial and more likely to affect families with unstable employment and lower annual income. Examining different strategies and interventions to reduce these costs such as virtual platforms, telephone and outreach clinics especially in poor communities and families with low annual income is warranted.

Funding Agencies

CIHRThe Children’s Hospital Research Institute of Manitoba