Molecular Parameters Research Articles

PATH-07. CLINICAL AND RADIOGRAPHIC FEATURES OF DIFFUSE GLIOMA MOLECULAR SUBTYPES IN ACCORDANCE WITH THE 2021 CNS5 WHO CLASSIFICATION

Abstract BACKGROUND The incorporation of molecular parameters into WHO CNS5 has led to reclassification of many gliomas. We undertook to describe presenting clinical and radiographic features of diffuse adult gliomas according to CNS5 category. METHODS We reviewed pathology reports, clinical and MRI data at presentation of 972 adult patients with glioma diagnosed between 1/2010-2/2022. Continuous variables were presented as median; categorical variables as numbers and percentages, and comparison were performed using chi-square test. RESULTS 736 patients had sufficient data for CNS5 re-classification and were included in the analysis: Grade 2 IDH-mutant (IDHmut) astrocytoma (A2) n=69, grade 3 IDHmut astrocytoma (A3) n=37, grade 4 IDHmut astrocytoma (A4), n=32, grade 2 oligodendroglioma (O2) n=60, grade 3 oligodendroglioma (O3) n=23, and grade 4 IDH-wildtype glioblastoma (GBM) n=515. Age at presentation significantly differed between grade 2&3 gliomas and grade 4 (mean age:39 versus 61.7; p<0.001); A4 were also younger than GBM (39.8 versus 63.1). A majority in all categories except grade 2 gliomas were male. Seizure was more common in IDHmut gliomas than GBM (58.4% versus 31.8%;p<0.001), while cognitive impairment was more common in GBM than in IDHmut gliomas (64.4% versus 34.4%;p<0.001) and in A4 (64% versus 40.6%;p=0.013). Focal deficits at presentation were more frequent in GBM than in IDHmut gliomas (74.2% versus 30.5%;p<0.001), but not significantly different compared to A4 (74.2% versus 58.1%;p=0.06). Contrast enhancement on MRI was more frequent in GBM compared to IDHmut gliomas (93% versus 48%;p<0.001), but similar frequency with A4 (93% versus 96%;p=1.00). The minimum ADC was significantly higher in IDHmut glioma including A4 than GBM(p<0.001). Calcification was more common in oligodendroglioma than astrocytoma and GBM (p<0.001), and more frequent in A4 than GBM(p=0.016). CONCLUSION Significant differences in the clinical presentation and radiologic features exist among CNS5 glioma subtypes, informing potential diagnosis, management and prognosis at initial presentation.

Impact of high-fructose diet and metformin on histomorphological and molecular parameters of reproductive organs and vaginal microbiota of female rat

There are limited data on the effects of a high-fructose diet on the female reproductive system. Although metformin has some functional effects on female fertility, its reproductive outcome on high fructose diet-induced metabolic syndrome is unclear. The aim of the present study is to evaluate the impact of a high fructose diet on histomorphological and molecular parameters of the reproductive organs and vaginal microbiota as well as the treatment potential of metformin. Wistar albino rats were used in the study. The metabolic syndrome model was induced by a high-fructose diet in rats for 15 weeks. Metformin was orally administered once a day for the last 6 weeks. The high-fructose diet increased blood glucose, triglycerides, insulin, and ovarian testosterone levels; however, it reduced ovarian aromatase levels and follicle numbers and caused uterine inflammation. The high-fructose diet-induced molecular abnormalities on ovarian tissue were demonstrated by the downregulation of ovarian insulin signaling pathway proteins and dysregulation of ovarian mitogenic and apoptotic pathway proteins. A high-fructose diet caused vaginal dysbiosis, metformin increased probiotic bacteria in the vaginal microbiota. Our results revealed that metformin improves ovarian impairments by modulating hormonal balance, insulin level, mapk, and apoptotic signaling molecules, as well as regulating the vaginal microbiota.

Determination the Anticorrosive Properties of Trimebutine Maleate for Aluminum Corrosion in 2M HCl Gravimetric and Quantum Chemistry Approaches

The present work evaluated the inhibition properties of (R,S)-2-(dimethylamino)-2-phenylbutyl 3,4,5-trimethoxybenzoate or trimebutine maleate (TM) in aluminum corrosion in 2M hydrochloric acid. Tests were carried out using gravimetric methods and density functional theory (DFT). The results indicate that trimebutine maleate is a good inhibitor of aluminum corrosion in 2M hydrochloric acid at low temperatures. Indeed, the inhibition efficiency of this molecule increases with increasing concentration and decreases with increasing temperature. For an inhibitor concentration of 5.2mM and at 298K, we obtained an inhibition efficiency of 93.61%. The adsorption process is spontaneous, exothermic and follows the Langmuir model. The activation parameters were calculated and analyzed. DFT at the B3LYP/6-311G (d.p) level was used to determine molecular parameters such as EHOMO, ELUMO, ΔE, η, S, μ , A, I, χ, ΔN, ω. These theoretical parameters showed that TM is highly reactive and can both donate and receive electrons from aluminum. They thus confirmed the good inhibition performance of the molecule studied. These parameters were used to explain the inhibition efficiencies obtained. Local selectivity was analyzed using Fukui functions and the dual descriptor to determine possible nucleophilic and electrophilic attack sites.

Low phosphorus causes hepatic energy metabolism disorder through Dynamin-related protein 1-mediated mitochondrial fission in fish

BackgroundLow phosphorus (LP) diets perturb hepatic energy metabolism homeostasis in fish. However, the specific mechanisms in LP-induced hepatic energy metabolism disorders remain to be fully elucidated. ObjectivesThis study sought to elucidate the underlying mechanisms of mitochondria involved in LP-induced energy metabolism disorders. MethodsSpotted seabass were fed diets with 0.72% (S-AP, control) or 0.36% (S-LP) available phosphorus for 10 weeks. Drp1 was knocked down or protein kinase A (PKA) was activated using 8Br-cAMP (5 μM, a PKA activator) in spotted seabass hepatocytes under LP medium. Zebrafish were fed Z-LP diets (0.30% available phosphorus) containing Mdivi-1 (5 mg/kg, a Drp1 inhibitor) or 8Br-cAMP (0.5 mg/kg) for 6 weeks. Biochemical and molecular parameters, along with transmission electron microscopy and immunofluorescence, were used to assess hepatic glycolipid metabolism, mitochondrial function and morphology. ResultsSpotted seabass fed S-LP diets showed reduced ATP (0.52-fold) and cyclic adenosine monophosphate (cAMP) (0.52-fold) levels, along with reduced Drp1 (s582) (0.38-fold) and PKA (0.61-fold) phosphorylation levels in the liver compared with those fed S-AP diets (P < 0.05). Drp1 knockdown elevated ATP levels (1.99-fold), decreased mitochondrial DRP1 protein levels (0.45-fold), and increased mitochondrial aspect ratio (1.82-fold) in LP-treated hepatocytes (P < 0.05). Furthermore, 8Br-cAMP-treated hepatocytes exhibited higher PKA phosphorylation (2.85-fold), ATP levels (1.60-fold), and mitochondrial aspect ratio (2.00-fold), along with decreased mitochondrial DRP1 protein levels (0.29-fold) under LP medium (P < 0.05). However, mutating s582 to alanine mimic Drp1 dephosphorylation decreased ATP levels (0.63-fold) and mitochondrial aspect ratio (0.53-fold) in 8Br-cAMP-treated hepatocytes (P < 0.05). In addition, zebrafish were fed Z-LP diets containing Mdivi-1 or 8Br-cAMP had higher ATP levels (3.44-fold or 1.98-fold) than that fed Z-LP diets (P < 0.05). ConclusionsThese findings provide a potential mechanistic elucidation for LP-induced energy metabolism disorders through the cAMP/PKA/Drp1-mediated mitochondrial fission signaling pathway.

Accumulation patterns and chronic toxic effects of triphenyl phosphine in Mytilus coruscus

Triphenylphosphine oxide (TPPO) as a new organophosphorus flame retardant is extensively used in the industrial and pharmaceutical fields due to its chemical stability and abundance. In marine environments, TPPO accumulates as a result of natural ecological processes. This study aimed to evaluate the accumulation process and toxicological effects of TPPO utilizing mussels (Mytilus coruscus), a species traditionally employed in marine monitoring and commonly cultivated in offshore aquaculture. GC–MS/MS was employed to quantify the accumulation of TPPO in the gill, gonad, muscle, and digestive gland tissues of mussels exposed to three sublethal concentrations (20.0, 100.0, and 500.0 μg/L) over durations of 3, 7, 14, and 28 days, as well as the examining of the chemical and molecular parameters. Results showed that the accumulation of TPPO (100.0 and 500.0 μg/L) increased throughout the 28 d of exposure, and the peak accumulation of 20.0 μg/L TPPO was detected on 14 d. Short-term TPPO exposure did not cause oxidative stress in mussels. Chronic exposure for 28 days resulted in significant oxidative damage and oxidative stress in mussels, as evidenced by elevated expression and activity of SOD, increased levels of glutathione and malondialdehyde, and upregulation of HSP70 and HSP90, metallothionein, and CYP4Y1, alongside the initiation of apoptotic cell death. TPPO exposure activated natural immune genes and disrupted lipid metabolism in mussels. This study demonstrated the persistent accumulation of TPPO in mussels, highlighting its ecotoxicological hazards and underscoring the necessity to predict the risks associated with widespread environmental exposure to TPPO.

Enhancing the differential diagnosis of small pulmonary nodules: a comprehensive model integrating plasma methylation, protein biomarkers, and LDCT imaging features

BackgroundAccurate differentiation between malignant and benign pulmonary nodules, especially those measuring 5–10 mm in diameter, continues to pose a significant diagnostic challenge. This study introduces a novel, precise approach by integrating circulating cell-free DNA (cfDNA) methylation patterns, protein profiling, and computed tomography (CT) imaging features to enhance the classification of pulmonary nodules.MethodsBlood samples were collected from 419 participants diagnosed with pulmonary nodules ranging from 5 to 30 mm in size, before any disease-altering procedures such as treatment or surgical intervention. High-throughput bisulfite sequencing was used to conduct DNA methylation profiling, while protein profiling was performed utilizing the Olink proximity extension assay. The dataset was divided into a training set and an independent test set. The training set included 162 matched cases of benign and malignant nodules, balanced for sex and age. In contrast, the test set consisted of 46 benign and 49 malignant nodules. By effectively integrating both molecular (DNA methylation and protein profiling) and CT imaging parameters, a sophisticated deep learning-based classifier was developed to accurately distinguish between benign and malignant pulmonary nodules.ResultsOur results demonstrate that the integrated model is both accurate and robust in distinguishing between benign and malignant pulmonary nodules. It achieved an AUC score 0.925 (sensitivity = 83.7%, specificity = 82.6%) in classifying test set. The performance of the integrated model was significantly higher than that of individual methylation (AUC = 0.799, P = 0.004), protein (AUC = 0.846, P = 0.009), and imaging models (AUC = 0.866, P = 0.01). Importantly, the integrated model achieved a higher AUC of 0.951 (sensitivity = 83.9%, specificity = 89.7%) in 5–10 mm small nodules. These results collectively confirm the accuracy and robustness of our model in detecting malignant nodules from benign ones.ConclusionsOur study presents a promising noninvasive approach to distinguish the malignancy of pulmonary nodules using multiple molecular and imaging features, which has the potential to assist in clinical decision-making.Trial registration: This study was registered on ClinicalTrials.gov on 01/01/2020 (NCT05432128). https://classic.clinicaltrials.gov/ct2/show/NCT05432128.

Clog P-Guided Development of Multi-Colored Buffering Fluorescent Probes for Super-Resolution Imaging of Lipid Droplet Dynamics.

Super-resolution fluorescence imaging of live cells increasingly demands fluorescent probes capable of multi-color and long-term dynamic imaging. Understanding the mechanisms of probe-target recognition is essential for the engineered development of such probes. In this study, it is discovered that the molecular lipid solubility parameter, Clog P, determines the staining performance of fluorescent dyes on lipid droplets (LDs). Fluorescent dyes with Clog P values between 2.5 and 4 can form buffering pools outside LDs, replacing photobleached dyes within LDs to maintain constant fluorescence intensity in LDs, thereby enabling dynamic super-resolution imaging of LDs. Guided by Clog P, four different colored buffering LD probes spanning the visible light spectrum have been developed. Using Structured Illumination Microscopy (SIM), the role of LD dynamics have been tracked during cellular ferroptosis with the secretion, storage, and degradation of overexpressed ACSL3 proteins. It is found that LDs serve as storage sites for these proteins through membrane fusion, and further degrade overexpressed proteins via interactions with organelles like lysosomes or through lipophagy, thereby maintaining cellular homeostasis.

Vibrational Spectra and Computational Study of Amyl Acetate: MEP, AIM, RDG, NCI, ELF and LOL Analysis

This work is focused on biologically active neat amyl acetate and its solutions in ethanol/heptane. According to the experimental results, when the concentration of amyl acetate in the amyl acetate-ethanol solution decreases, the additional band appears on the low-frequency side. The primary reason for the formation of such additional band is the intermolecular hydrogen bonding between amyl acetate and ethanol. In the amyl acetate-heptane solution, as the concentration of amyl acetate in the solution decreases, the band corresponding to the C=O stretching vibrations shifted to a higher frequency. This is explained by the fact that heptane breaks intermolecular interactions in solution, resulting in a simpler spectral band corresponding to the C=O stretching vibrations. Calculations are also used to study interactions in amyl acetateethanol complexes and their spectral manifestations. When the complex formation energies are calculated, this energy increases with the number of molecules, but the average hydrogen bond energy per one bond remains unchanged. The density functional theory (DFT) method is used to analyze molecular structural parameters: Mulliken atomic charge distribution; thermodynamic parameters; molecular electrostatic potential (MEP) surface; atoms in molecules (AIM) analysis; quantum chemical parameters such as reduced density gradient (RDG) and noncovalent interaction (NCI) analysis; electron localization functions (ELF) analysis; and localized orbital locator (LOL) analysis.

Cancer ameliorates cardiac dysfunction and suppresses fibrosis in a mouse model for Duchenne muscular dystrophy

Abstract The interplay between heart failure and cancer represents a double-edged sword. Whereas cardiac remodeling promotes cancer progression, tumor growth suppresses cardiac hypertrophy and reduces fibrosis deposition. Whether these two opposing interactions are connected awaits to be determined. In addition, it is not known whether cancer affects solely the heart, or if other organs are affected as well. To explore the dual interaction between heart failure and cancer, we studied the human genetic disease Duchenne Muscular Dystrophy (DMD) using the MDX mouse model. We analyzed fibrosis and cardiac function as well as molecular parameters by multiple methods in the heart, diaphragm, lungs, skeletal muscles, and tumors derived from MDX and control mice. Surprisingly, cardiac dysfunction in MDX mice failed to promote murine cancer cell growth. In contrast, tumor-bearing MDX mice displayed reduced fibrosis in the heart , skeletal and diaphragm muscles, resulting in improved cardiac function. The latter is at least partially mediated via M2 macrophage recruitment to the heart and diaphragm muscles. Remarkably, an analysis of of cytokines in the blood of tumor-bearing and non-tumor-bearing MDX mice initially revealed several potential factors. These factors hold promise in significantly reducing fibrosis within our model. Administering these factors simultaneously to MDX mice led to decreased fibrosis in the heart, diaphragm, and skeletal muscles, along with an improvement in cardiac function. Collectively, our data support the notion that the effect of heart failure on tumor promotion is independent of the improved cardiac function in tumor-bearing mice. Reduced fibrosis in tumor-bearing MDX mice stems from the suppression of new fibrosis synthesis and the removal of existing fibrosis. These findings offer potential therapeutic strategies for DMD patients, fibrotic diseases, and cardiac dysfunction.Tumor growth suppresses fibrosisGraphical representation

Individual and combined effects of commercial glyphosate, atrazine and 2,4-D herbicides on the gerbil ventral prostate

Exposure to pesticides, individually or in a mixture, in drinking water is one of the main sources of human contamination, which causes adverse effects on the reproductive system. Our study aimed to investigate, the effects of a 90-day exposure to low concentrations of glyphosate (GLY), atrazine (ATZ), and 2,4-dichlorophenoxyacetic acid (2,4-D), in commercial formulations, on morphological, molecular, and hormonal parameters of the ventral prostate of gerbils (Meriones unguiculatus). The animals were exposed via drinking water to individual concentrations of GLY: 700 μg/L, ATZ: 3 μg/L, and 2,4-D: 70 μg/L, as well as to their mixture (MIX). Our findings showed an increase in prostatic complex relative weight in ATZ-exposed animals. Stereological and morphometric techniques indicated an increase in the percentage and thickness of muscular stroma, following an increase in the amount of collagen and reticular fibers in the MIX group. Histopathological analysis showed a decrease in the incidence of epithelial atrophy, subepithelial inflammation, and microacini in the MIX. On the other hand, ATZ-exposed animals showed an increase in hyperplasia and total prostatic intraepithelial neoplasia (PIN). The expression of caspase-3 decreased and estrogen receptor alpha (ERα) increased in the 2,4-D and MIX. Western blotting showed an increase in estrogen receptor beta (ERβ) expression in MIX-exposed animals. Testosterone levels decreased in animals from the GLY, ATZ and 2,4-D groups. Our findings provide evidence that individual or combined exposure to herbicides causes hormonal imbalance and morphological alterations, besides favoring the incidence of proliferative lesions in the prostate, predisposing the gland to more severe injuries.

Intracavernosal mesenchymal stem cell therapy in ischaemic priapism: an experimental study.

The most common form of priapism is ischaemic and its prevalence in men has increased in recent years as a result of intracavernosal drug use. Currently, there is no approved specific treatment for ischaemic priapism other than cavernosal aspiration, which can only provide detumescence. This study aims to evaluate the efficacy of intracavernosal mesenchymal stem cell (MSC) therapy in an ischaemic priapism model. Thirty male Wistar albino rats were divided into three groups: sham (n = 6), priapism (n = 12) and priapism + MSC treatment (n = 12). The experimental groups were also divided into 1 and 12h subgroups of ischaemic priapism. The experimental model was created using a vacuum erection device and constrictive tape technique, and intracavernosal MSC were applied immediately after the tape was removed. After 4weeks, intracavernosal pressures (ICPs) and systemic mean arterial pressure (MAP) were measured. Penectomy was then performed to assess histopathological and molecular changes in the rats' penile tissues. In the ischaemic priapism model, MSC therapy showed significant improvements in peak and mean ICPs and mean ICP/MAP ratio. Histopathological analysis showed significant increases in smooth-muscle/collagen ratio and e-NOS and n-NOS expression. Although there was a decrease in fibrosis, it was not significant. At the molecular level, there were significant decreases in TGF-beta and VEGF mRNA expression, whilst NGF and BDNF mRNA-expression levels showed significant increases with MSC therapy. In terms of ICPs, the therapy showed more significant improvements in short-term priapism. However, when looking at histopathological and molecular parameters, the therapy had positive effects on a wider range of parameters in the long-term priapism. MSC treatment improved cavernosal physiology and had positive effects at the histopathological and molecular level in the ischaemic priapism model.

Analysis of Molecular Imaging Biomarkers Derived from [18F]FDG PET/CT in mCRPC: Whole-Body Total Lesion Glycolysis (TLG) Predicts Overall Survival in Patients Undergoing [225Ac]Ac-PSMA-617-Augmented [177Lu]Lu-PSMA-617 Radioligand Therapy

Background/Objectives: The augmentation of [177Lu]Lu-PSMA-617 radioligand therapy by alpha emitting [225Ac]Ac-PSMA-617, known as the tandem therapy concept, is a promising escalating treatment option in advanced mCRPC. In this study, we evaluated the value of [18F]FDG PET/CT-derived molecular imaging biomarkers for predicting response and outcome to PSMA tandem RLT in n = 33 patients with insufficient response on [177Lu]Lu-PSMA-617 monotherapy. Methods: Six different molecular imaging parameters at baseline, i.e., before initiation of PSMA tandem RLT with respect to SUVmax, SUVpeak, SUV5, SUVmean, metabolic tumor volume (MTV), and total lesion glycolysis (TLG) were tested for association with response and overall survival (OS). Results: After the initiation of augmentation, 24.2% of patients with a previously insufficient response experienced partial remission, and 39.4% experienced stable disease. The median OS was 7 months (95% CI: 4–11 months). None of the tested parameters were able to predict the response (all p &gt; 0.529). In contrast, the [18F]FDG PET/CT-derived whole-body molecular imaging parameter TLG was significantly (p = 0.029) associated with OS of patients undergoing [225Ac]Ac-PSMA-617 augmented [177Lu]Lu-PSMA-617 RLT after insufficient response to [177Lu]Lu-PSMA-617 monotherapy. Conclusion: Implementing [18F]FDG PET/CT in the management of PSMA-RLT in clinical practice may contribute to outcome prediction and provide a route to more individualized management in mCRPC.

Chlorpyrifos and dimethoate exposure impairs female fertility by deregulating WNT signaling pathway & uterine receptivity

The study assessed histological, biochemical, oxidative stress, and molecular parameters to evaluate the consequences of Chlorpyrifos and Dimethoate exposure on uterine health in female rats. Despite showing no obvious signs of toxicity apart from minor clinical symptoms in DM-exposed rats, both pesticides caused degenerative changes in uterine tissue. This study demonstrates that pesticides induce inflammatory responses and oxidative stress in rats, by NF-κB activation and altering antioxidant enzyme levels. Besides, CPF and DM exposure disrupted gene expression of HOXA10, HOXA11, and WNT and reduced activation of β-catenin in the uterus, which is crucial for implantation and reproductive function. These findings suggest that pesticide exposure may impair reproductive health and fertility in females, highlighting potential implications for human health.

Rheological analysis of network structure of raw natural rubber prepared by different processing techniques

The performance of raw natural rubber (NR) is dominated by its network structure, particular the levels of long chain branching (LCB) and entanglement. Here, three type of raw rubbers were prepared using different processing methods for commercial grade NRs. Linear and nonlinear viscoelastic behaviors, as well as stress relaxation, were analyzed to access their network structures. The third relative harmonic, phase angle, and first to second quarter-period integral ratio of stress curve were utilized as indicators for the nonlinearity of samples. By combining these values with flow activation energy and characteristic times, it can be confirmed that naturally coagulated NR showed higher elasticity than acid-coagulated NR due to high levels of LCB and entanglement, and hot air drying could lead to chain degradation. These findings correlated with molecular weight parameters, gel content and Mooney viscosity results. This research establishes a method for monitoring raw NR quality and predicting its properties.

8509 Molecular and Reproductive Assessment in Ames strain under or not Hormonal Replacement

Abstract Disclosure: B.V. Azevedo: None. N. Trigueiro: None. J.M. Marques: None. V. Yariwake: None. L. Tamashiro: None. R. Cruz: None. M.M. Veras: None. L. Carvalho: None. Introduction: The Ames lineage, a mouse model with a mutation in the Prop1 gene, exhibits GH, TSH, and PRL deficiencies along with low levels of gonadotropins. This has prompted an investigation into aging and reproductive restoration after hormone replacement reported in the literature. Aims: The present study aims to analyze the pathophysiologic mechanisms in the reproductive system and fertility restoration. Methods: This is a prospective study approved by the ethical committee for animal use at the University of Sao Paulo Medical School. An isogenic Ames lineage was categorized into four groups: 1) Ames wild-type mice, 2) untreated Ames dwarf mice with sexual maturity, 3) untreated Ames dwarf mice without sexual maturity, and 4) Ames dwarf mice under hormonal treatment. Male Ames dwarf mice received treatment with levothyroxine and human recombinant GH, starting at 30 days postnatal (dp) for 40 days, followed by weekly applications until 90 dp. At the time of euthanasia, a set of analyses were conducted on animals from each group to characterize sexual maturation, fertility, and tissue collection for reproductive parameter analyses, such as seminiferous tubule morphology by histology, testicular weight, gonadosomatic index, and spermogram analysis. Additionally, pituitary RNAseq was performed using the poly-A tail capture method with the Illumina stranded mRNA prep kit and sequenced on the NextSeq 2000. Results: Treated dwarf mice exhibited fertility restoration, an increase in testicular volume with an improvement of testicular function, and complete spermatogenesis. Untreated Ames dwarf mice were infertile, but those that reached sexual maturity without hormonal replacement showed improvements in reproductive parameters, such as enhanced testicular development, maturation of some seminiferous tubules, increased testicular weight, and improvement in the spermogram compared to immature Ames dwarf mice. Transcriptome analyses revealed distinct patterns of all Ames dwarf mice groups and the wild-type. It was observed a decrease in Fshb expression in the group without sexual maturation compared to the wild-type group. Additionally, Prl and Slc6a12 were upregulated in the spontaneously sexually mature Ames dwarf compared to the non-restored Ames dwarf mice. Conclusion: GH and levothyroxine treatment in the Ames lineage are effective in restoring fertility and improving reproductive parameters in male Ames dwarf mice. However, sexually mature Ames dwarf mice without treatment presented improvements in reproductive and molecular parameters despite remaining infertile. These findings provide valuable insights into the complex interaction among hormonal replacement, modulating mechanisms, and sexual maturity. Presentation: 6/2/2024

8509 Molecular and Reproductive Assessment in Ames strain under or not Hormonal Replacement

Abstract Disclosure: B.V. Azevedo: None. N. Trigueiro: None. J.M. Marques: None. V. Yariwake: None. L. Tamashiro: None. R. Cruz: None. M.M. Veras: None. L. Carvalho: None. Introduction: The Ames lineage, a mouse model with a mutation in the Prop1 gene, exhibits GH, TSH, and PRL deficiencies along with low levels of gonadotropins. This has prompted an investigation into aging and reproductive restoration after hormone replacement reported in the literature. Aims: The present study aims to analyze the pathophysiologic mechanisms in the reproductive system and fertility restoration. Methods: This is a prospective study approved by the ethical committee for animal use at the University of Sao Paulo Medical School. An isogenic Ames lineage was categorized into four groups: 1) Ames wild-type mice, 2) untreated Ames dwarf mice with sexual maturity, 3) untreated Ames dwarf mice without sexual maturity, and 4) Ames dwarf mice under hormonal treatment. Male Ames dwarf mice received treatment with levothyroxine and human recombinant GH, starting at 30 days postnatal (dp) for 40 days, followed by weekly applications until 90 dp. At the time of euthanasia, a set of analyses were conducted on animals from each group to characterize sexual maturation, fertility, and tissue collection for reproductive parameter analyses, such as seminiferous tubule morphology by histology, testicular weight, gonadosomatic index, and spermogram analysis. Additionally, pituitary RNAseq was performed using the poly-A tail capture method with the Illumina stranded mRNA prep kit and sequenced on the NextSeq 2000. Results: Treated dwarf mice exhibited fertility restoration, an increase in testicular volume with an improvement of testicular function, and complete spermatogenesis. Untreated Ames dwarf mice were infertile, but those that reached sexual maturity without hormonal replacement showed improvements in reproductive parameters, such as enhanced testicular development, maturation of some seminiferous tubules, increased testicular weight, and improvement in the spermogram compared to immature Ames dwarf mice. Transcriptome analyses revealed distinct patterns of all Ames dwarf mice groups and the wild-type. It was observed a decrease in Fshb expression in the group without sexual maturation compared to the wild-type group. Additionally, Prl and Slc6a12 were upregulated in the spontaneously sexually mature Ames dwarf compared to the non-restored Ames dwarf mice. Conclusion: GH and levothyroxine treatment in the Ames lineage are effective in restoring fertility and improving reproductive parameters in male Ames dwarf mice. However, sexually mature Ames dwarf mice without treatment presented improvements in reproductive and molecular parameters despite remaining infertile. These findings provide valuable insights into the complex interaction among hormonal replacement, modulating mechanisms, and sexual maturity. Presentation: 6/2/2024

Molecular insights into 5-hydroxymethylfurfural: a computational, spectroscopic, and docking investigation

The quantum chemical properties of 5-hydroxymethylfurfural were investigated using Density Functional Theory alongside vibrational spectroscopy. Key outcomes included optimizing molecular structure, vibrational frequencies, and various molecular parameters. By comparing DFT results with experimental infrared spectra, molecular motion was clarified. Reactive sites were identified through Molecular Electrostatic Potential and Fukui function analyses. Hirshfeld surface analysis revealed insights into the crystal structure’s intermolecular interactions and hydrogen bonding. Time-dependent Density Functional Theory combined with the Polarizable Continuum Model provided Ultraviolet spectra, highlighting charge transfer between the highest occupied and lowest unoccupied molecular orbitals. The compound’s electronegativity (4.7239) and electron affinity were assessed. Biological studies, including drug-likeness evaluations and molecular docking, also demonstrated potential physiological benefits, mainly through the compound’s low binding energy. A 100-nanosecond molecular dynamics simulation of the 5-HMF-4LB4 complex revealed its stability and dynamic behavior through analyses of Root Mean Square Deviation, Root Mean Square Fluctuation, hydrogen bonding, Solvent Accessible Surface Area, and radius of gyration.

Characteristics of risk factors and molecular genetic parameters in children with chronic kidney disease

Introduction. Information about the health status of children with nephrological pathology includes many manifestations of functional system disorders. However, early diagnosis of chronic kidney disease (CKD) is rarely carried out due to the lack of reliable predictors of this process.The aim: To analyze the association of anamnestic signs, laboratory and genetic research results with chronic kidney disease in children to identify potential predictors of the disease.Material and Methods. The data were obtained from a single-center retrospective catamnestic cohort study (2011–2022). The main group included 128 children with chronic kidney disease aged 1 to 17 years. The comparison group consisted of 30 children without diagnosed kidney pathology aged from 1 to 17 years. There were no statistically significant differences in sex and age between the two groups. The informative value of more than 150 features, including anamnestic ones, the state of the mother’s health during pregnancy, antenatal and perinatal factors, features of the child’s development and manifestation of the disease, the results of clinical, laboratory, and genetic research methods were evaluated. An integrated approach to the study included methods for hypotheses testing on statistical significance of indicators differences, regression analysis, and ROC analysis.Results. Hereditary burden of nephrological pathology in the family, unfavorable course of pregnancy (anemia, toxicosis of the first half), obstetric and somatic pathology in the mother, the serial number of childbirth are associated with the subsequent development of CKD. Erythrocytes in the urine and daily protein loss were detected only in the group of children with CKD. In this group, a decrease in the number of red blood cells, increased erythrocyte sedimentation rate (ESR), allergies at an early age, otitis media, persistent viral and bacterial infections, and a number of signs of connective tissue dysplasia were more common. The main group statistically significantly differed from the comparison group by a number of polymorphic genetic markers. It can be assumed that the polymorphisms AGT1 Thr174Thr, AGT2 Thr235Thr, NO3 C786T are associated with a predisposition to CKD, and the products of polymorphisms AGT1 Thr174Met, AGT2 Met235Met, R1 AGT2 C1166C, NO3 T786T may have a protective effect according to development of CKD.Conclusion. The risk factors for CKD identified in this study can be used to develop software to support medical decision making for early detection of children at high risk of disease progression.

A density functional theory study of a series of symmetric dibenzylideneacetone analogues as potential chemical UV-filters

The aim of this research was to provide valuable insights on symmetrical α,β-unsaturated ketones as potential chemical ultraviolet (UV) filters from experimental data and theoretical aspects. Towards this end, density functional theory (DFT/B3LYP) calculations on a series of symmetrical α,β-unsaturated ketones, (1E,4E)-1,5-bis[4-(R)phenyl]penta-1,4-diene-3-one (R = methylthio, 1; R = dimethylamino, 2; R = ethyl, 3), were performed to determine the effect of different electron-donating substituents on their stability when exposed to solar UV radiation. Their molecular structures, and UV-visible, infrared (IR) and NMR (1H and 13C) spectra were theoretically obtained from their optimized geometries with the B3LYP/6-311++ G (d, p) basis set and were compared with the experimental results. Conformational analysis was performed and the most stable conformer of each compound was identified as the trans-trans conformer, which was further supported by experimental NMR data. The UV spectra and effect of solvent polarity and proticity were studied by the time-dependent-DFT (TD-DFT) approach with the B3LYP/6-311++G (d, p) level of theory. Furthermore, various molecular parameters like dipole moment, frontier molecular orbital (FMO) energies, ΔEHOMO-LUMO gap, lifetime of the first excited state (τ), global chemical reactivity descriptors, and natural bond orbital analysis were predicted at the same level of theory and compared with the experimental data. Inspection of the active FMOs revealed the photoinstability trends of 1 and 3 under UV irradiation. However, introducing a -N(CH3)2 substituent to 2 at the para-position improves its photostability towards simulated UV radiation. Thus, compound 2 has the potential to provide efficient broad-spectrum protection against UV radiation. This work has shown that molecular modelling strategies can assist to rationalize experimental findings and also support the identification of photoproducts of 1 and 3.

Antiquorum Sensing Effect, Theoretical, and X‐Ray Studies of a Schiff Base Molecule Containing Pyrimidine Nucleus

AbstractCompound 4, a Schiff base‐containing pyrimidine moiety, was synthesized from the reaction of N‐amino pyrimidine derivative with 4‐fluorobenzaldehyde. The molecular and crystallographic structure of 4 was determined by the SC‐XRD method. According to these results, compound 4 crystallized in the triclinic P‐1 space group, and the stability of its crystal structure was ensured through intermolecular C−H⋅⋅⋅π and π⋅⋅⋅π interactions. In addition to the antibacterial effects of the molecule on some Gram‐positive and Gram‐negative bacteria, the inhibition of biofilm formation in P. aeruginosa PAO1 and the production of violacein pigment in Chromobacterium violaceum (C. violaceum) 12472 were studied. The results demonstrated that compound 4 had strong effectiveness in preventing the formation of biofilms produced by PAO1 with an inhibition rate of 69 %. Extensive theoretical calculation studies of molecule have been conducted and the molecular structure of 4 has been optimized in the ground state using density functional theory (DFT). The investigated compound's frontier molecular orbitals, chemical parameters, and molecular electrostatic potential surfaces were examined. Molecular docking studies were performed to explain the binding interaction 4 with the high inhibitory effect against C. violaceum phenylalanine hydroxylase D139A (PDB ID: 4Q3Y), D139E (PDB ID: 4Q3W), D139K (PDB ID: 4Q3Z), and D139N (PDB ID: 4Q3X) mutations.