ATF2 loss promotes 5-FU resistance in colon cancer cells via activation of the ATR-Chk1 damage response pathway.

BACKGROUND: The role of ATF2 in colon cancer (CC) is controversial. Recently, we reported that low ATF2 expression is characteristic of highly invasive tumors, suggesting that ATF2 might also be involved in therapy resistance. 5-Fluorouracil (5-FU) is the best-known chemotherapeutic drug for CC, but drug resistance affects its curative effect. To date, the role of ATF2 in the 5-FU response remains elusive. METHODS/RESULTS: For our study, we had available HCT116 cells (wild-type p53) and HT29 colon tumor cells (mutant p53) and their corresponding CRISPR‒Cas9-generated ATF2-KO clones. We observed that loss of ATF2 triggered dose- and time-dependent 5-FU resistance in HCT116 cells by activating the DNA damage response (DDR) pathway with high p-ATRThr1989 and p-Chk1Ser317 levels accompanied by an increase in the DNA damage marker γ-H2AX in vitro and in vivo using the chicken chorioallantoic membrane (CAM) model. Chk1 inhibitor studies causally displayed the link between DDR and drug resistance. There were contradictory findings in HT29 ATF2-KO cells upon 5-FU exposure with low p-Chk1Ser317 levels, strong apoptosis induction, but no effects on DNA damage. In ATF2-silenced HCT116 p53-/- cells, 5-FU did not activate the DDR pathway. Co-immunoprecipitation and proximity ligation assays revealed that upon 5-FU treatment, ATF2 binds to ATR to prevent Chk1 phosphorylation. Indeed, in silico modelling showed reduced ATR-Chk1 binding when ATF2 was docked into the complex. CONCLUSIONS: We demonstrated a novel ATF2 scaffold function involved in the DDR pathway. ATF2-negative cells are highly resistant due to effective ATR/Chk1 DNA damage repair. Mutant p53 seems to overwrite the tumor suppressor function of ATF2.

Optimised Protocol for Managing Failed Catheterisation: Leveraging Bedside Retrograde Urethrography and Cystourethroscopy.

Introduction Urethral catheterization is a routine procedure often required for many hospitalized patients. Various conditions, such as meatal stenosis, stricture urethra, false passage, benign prostatic hyperplasia, bladder neck contractures, and impacted urethral stones, can contribute to difficulty in catheterisation. In the setting of failed attempts at per urethral catheter placement, the subsequent intervention is suprapubic catheter (SPC) insertion. SPC placement has its associated complications and causes inconvenience to the patients. We framed an algorithm to minimise the need for SPC insertion in cases of difficult per urethral catheterisation in a non-trauma setting. This study aimed to evaluate the common causes of difficult per urethral catheterisation and establish the efficacy of our algorithm in managing difficult catheterisation with bedside retrograde urethrography (RGU) and cystoscopy while avoiding SPC placement. Materials and methods This prospective observational study was conducted from September 2022 to June 2024. Patients admitted with urinary retention or requiring routine catheterisation, with one failed attempt at catheterisation, were included in the study. Our algorithm for the management of difficult catheterisation in a non-trauma setting, to avoid SPC, integrates a bedside RGU and retrograde urethroscopy using either a 15.5 Fr cystoscope sheath or a 6 Fr ureteroscope to identify the urethral pathology, followed by dilatation and per urethral catheterisation. Results Among 55 patients (aged 34-82 years), 48 (87.27%) were male and seven (12.73%) were female. The most common indication for catheterisation was routine catheterisation for output monitoring (n = 30; 54.54%), followed by acute retention (n = 25; 45.45%). Bulbar urethral stricture (n = 28; 50.9%) was the most common cause of difficult catheterisation, followed by meatal/sub-meatal narrowing (n = 13; 23.63%), enlarged prostate or high bladder neck (n = 4; 7.27%), and impacted stones (n = 3; 5.45%). Successful catheterisation was achieved in 48 male patients following urethroscopy with a 6 Fr ureteroscope or 15.5 Fr cystoscope. In females, reducing the pelvic organ prolapse enabled catheterisation in two cases, while five required serial dilatation and catheterisation. Successful per-urethral catheterisation was achieved in all 55 (100%) patients, thus avoiding SPC. Conclusions Conventional blind catheterisation techniques have limited success in the setting of failed initial catheterisation. This approach, which employs bedside fluoroscopy and direct visualisation of the urethra using a cystoscope or ureteroscope, helped achieve higher success rates (n = 55; 100%) for difficult per-urethral catheterisation and avoided the need for SPC. Proper implementation of this protocol for dealing with difficult per-urethral catheterisation will reduce the unnecessary burden on the healthcare system by minimising the potential iatrogenic urethral injuries and reducing the need for SPC.

Structural Characterization, Spectroscopic Profile, Molecular Docking, ADMET Properties, Molecular Dynamics Simulation Studies, and Molecular Mechanics Generalized Born Surface Area Analysis of 5-(Adamantan-1-yl)-4-butyl-2,4-dihydro-3H-1,2,4-triazole-3-thione as a Potential COX Inhibitor.

Employing a synergistic combination of theoretical density functional theory (DFT) and experimental techniques, we conducted a comprehensive analysis elucidating the structural and pharmacological attributes of 5-(adamantan-1-yl)-4-butyl-2,4-dihydro-3H-1,2,4-triazole-3-thione (5A4BT) as a potent COX inhibitor. The X-ray crystallographic data of 5A4BT showed the pivotal role played by weak interactions, notably π-π and C-H-π interactions, alongside hydrogen bonding, in orchestrating the intricate supramolecular architectures within the crystalline lattice. A quantitative analysis of the arrangement of the crystal structure, as well as both inter- and intramolecular interactions, was conducted using Hirshfeld surfaces and 2D fingerprint plots. Additionally, a comprehensive examination of the IR spectra was undertaken, employing both experimental methods and theoretical DFT techniques, to elucidate the vibrational characteristics of the compound. The strength of intermolecular N-H···S hydrogen bonding and charge transfer within the system was assessed through natural bonding orbital analysis. Moreover, Bader's atoms in molecules theory was employed to estimate the strength of intermolecular hydrogen bonds, revealing strong interactions within the 5A4BT dimer. The title compound exhibited binding affinities of -6.4 and -6.5 kcal/mol for COX1 (PDB 3KK6) and COX2 (1CX2) target proteins, respectively. For the first time, predictions regarding ADMET properties, drug-likeness, and toxicity, including favorable bioavailability, along with 100 ns molecular dynamics simulations, binding free energy, and energy decomposition per residue in the binding cavity of the protein from molecular mechanics generalized born surface area approach, collectively indicate the potential of 5A4BT as a nonselective COX inhibitor.

Histone variants shape chromatin states in Arabidopsis.

How different intrinsic sequence variations and regulatory modifications of histones combine in nucleosomes remain unclear. To test the importance of histone variants in the organization of chromatin we investigated how histone variants and histone modifications assemble in the Arabidopsis thaliana genome. We showed that a limited number of chromatin states divide euchromatin and heterochromatin into several subdomains. We found that histone variants are as significant as histone modifications in determining the composition of chromatin states. Particularly strong associations were observed between H2A variants and specific combinations of histone modifications. To study the role of H2A variants in organizing chromatin states we determined the role of the chromatin remodeler DECREASED IN DNA METHYLATION (DDM1) in the organization of chromatin states. We showed that the loss of DDM1 prevented the exchange of the histone variant H2A.Z to H2A.W in constitutive heterochromatin, resulting in significant effects on the definition and distribution of chromatin states in and outside of constitutive heterochromatin. We thus propose that dynamic exchanges of histone variants control the organization of histone modifications into chromatin states, acting as molecular landmarks.

Spectroscopic, electronic structure, molecular docking, and molecular dynamics simulation study of 7-Trifluoromethyl-1H-indole-2-carboxylic acid as an aromatase inhibitor.

The present work encompasses a combined experimental and theoretical investigation of the molecular structure, vibrational wavenumbers, electronic structure at the ground and electronic excited states, molecular electrostatic potential surface of 7-(Trifluoromethyl)-1H-indole-2-carboxylic acid (TICA) and possibility of the title molecule as an aromatase inhibitor using molecular docking and molecular dynamic simulations. A stable conformer has been obtained using potential energy scans by varying appropriate dihedral angles. The obtained minimum energy conformer was further optimized at the 6-311++G (d, p) basis set by applying the most accepted B3LYP functional. A good agreement between experimental and calculated normal modes of vibration has been observed. The hydrogen-bonded interaction between two monomeric units of TICA has been investigated using NBO,QTAIM, and NCI (noncovalent interactions) analysis. Molecular docking of TICA with human placental aromatase (PDB ID: 3S79) reveals the formation of polar hydrogen bonds as well as hydrophobic interactions between the ligand and the protein, right in the binding cavity. TICA satisfies all pharmacokinetic filters (Lipinski rule of five, the Veber rule, Ghose rule, Egan rule, as well as the Muegge rule) and has a high bioavailability score of 0.85. Dynamic stability of the ligand within the binding pocket of the target protein has been confirmed by 100 ns molecular dynamics simulation results. The present study provides an excellent starting point for additional in vivo research, and TICA may eventually serve as a significant therapeutic candidate for the treatment of breast cancer.

Use of some cost-effective technologies for a routine clinical pathology laboratory.

Classically, the need for highly sophisticated instruments with important economic costs has been a major limiting factor for clinical pathology laboratories, especially in developing countries. With the aim of making clinical pathology more accessible, a wide variety of free or economical technologies have been developed worldwide in the last few years. 3D printing and Arduino approaches can provide up to 94% economical savings in hardware and instrumentation in comparison to commercial alternatives. The vast selection of point-of-care-tests (POCT) currently available also limits the need for specific instruments or personnel, as they can be used almost anywhere and by anyone. Lastly, there are dozens of free and libre digital tools available in health informatics. This review provides an overview of the state-of-the-art on cost-effective alternatives with applications in routine clinical pathology laboratories. In this context, a variety of technologies including 3D printing and Arduino, lateral flow assays, plasmonic biosensors, and microfluidics, as well as laboratory information systems, are discussed. This review aims to serve as an introduction to different technologies that can make clinical pathology more accessible and, therefore, contribute to achieve universal health coverage.

Downmodulation of lysophosphatidic acid by Berberine loaded folate-conjugated glycol chitosan nanoparticles (BFGCN) to mitigate Rheumatoid arthritis (RA) & Cardio-vascular disease(CVD): Current knowledge and future perspectives

The perils of cardiovascular diseases (CVD) are enhanced by systemic chronic inflammation in autoimmune disorders like Rheumatoid arthritis (RA), in which the patients generally exhibit a high inflammatory burden, dyslipidemia causing 50-60% of RA patients susceptible to CVD dependent mortality. Lysophosphatidic acid (LPA) is a polar, pleiotropic lipid molecule that is water soluble and present in the synovial fluid that can be exploited as an effective biomarker for lipid-signalling. Current research on alternative medicine has recognized various new molecular targets of Berberine (BBR) and established novel signals in support of the efficacy and therapeutic potential of BBR to fight CVD. Therefore, BBR, an alkaloid with poor aqueous solubility could be foreseen as a therapeutic strategy for the reduction of inflammation induced lipidemia by targeting the macrophages and modulating their functions. Hence, a novel BBR loaded folate-conjugated glycol chitosan nanoparticles (BFGCN) could be hypothesized as a three-pronged approach to target activated macrophages, fibroblasts of synovial fluid for downmodulation of LPA. The greatest challenge is the heterogeneity, complexity and interdependence of RA and CVD. Investigation of prognostic and predictive biomarkers is urgently required. Therefore, an improved understanding of the pathogenesis of RA would facilitate identifying an improved targeted treatment and management of RA patients.

Study on molecular structure, spectroscopic behavior, NBO, and NLO analysis of 3-methylbezothiazole-2-thione.

Experimentally observed spectral data (FT-TR and FT-Raman) of 3-methylbezothiazole-2-thione (3MBT2T) were compared with the spectral data obtained by DFT/B3LYP method using 6-311++G(d,p) basis set. UV-Vis spectrum of the title compound was recorded and the electronic properties, such as frontier molecular orbitals and band gap energies were calculated by TD-DFT approach. The molecular properties like dipole moment, polarizability, first static hyperpolarizability, molecular electrostatic potential surface (MEPs), and contour map were calculated to get a better comprehension of the properties of the title molecule. Natural bond orbital (NBO) analysis was applied to investigate the stability of the molecule arising from charge delocalization. Global and local reactivity descriptors were also computed to predict reactivity and reactive sites on the molecule.

FT-IR, FT-Raman, NMR, UV and quantum chemical studies on monomeric and dimeric conformations of 3,5-dimethyl-4-methoxybenzoic acid.

Extensive spectroscopic investigations along with theoretical quantum chemical studies on 3,5-dimethyl-4-methoxybenzoic acid (DMMBA) have been consummated. The fundamental vibrational transitions were addressed by experimental FT-IR (4000-400cm(-1)) and FT-Raman (4000-10cm(-1)) techniques and density functional calculations at B3LYP/6-311++G(d,p) and B3LYP/6-311++G(df,pd) levels of theory. The (1)H, (13)C and DEPT 135 NMR spectra of studied compound were recorded in deuterated dimethylsulfoxide (DMSO-d6), and compared with computed data obtained by using gauge including atomic orbital (GIAO) method. The electronic absorption spectra in methanol and ethanol solution were evaluated in the range of 200-400nm, and TD-DFT method was chosen for computational study. The spectroscopic and theoretical results were compared to the corresponding properties for monomer and dimer structures for the most stable conformer. Stability of the molecule arising from hyperconjugative interactions and charge delocalization has been analyzed using natural bond orbital (NBO) analysis. Moreover, the thermodynamic and nonlinear optical (NLO) properties were evaluated.

Spectroscopic (FT-IR, FT-Raman, and UV-visible) and quantum chemical studies on molecular geometry, Frontier molecular orbitals, NBO, NLO and thermodynamic properties of 1-acetylindole.

Quantum chemical calculations of ground state energy, geometrical structure and vibrational wavenumbers of 1-acetylindole were carried out using density functional (DFT/B3LYP) method with 6-311++G(d,p) basis set. The FT-IR and FT-Raman spectra were recorded in the condensed state. The fundamental vibrational wavenumbers were calculated and a good correlation between experimental and scaled calculated wavenumbers has been accomplished. Electric dipole moment, polarizability and first static hyperpolarizability values of 1-acetylindole have been calculated at the same level of theory and basis set. The results show that the 1-acetylindole molecule possesses nonlinear optical (NLO) behavior with non-zero values. Stability of the molecule arising from hyper-conjugative interactions and charge delocalization has been analyzed using natural bond orbital (NBO) analysis. UV-Visible spectrum of the molecule was recorded in the region 200-500nm and the electronic properties like HOMO and LUMO energies and composition were obtained using TD-DFT method. The calculated energies and oscillator strengths are in good correspondence with the experimental data. The thermodynamic properties of the compound under investigation were calculated at different temperatures.

Cerebral Hydatid Cyst: A Case Report.

Objective: Cerebral Hydatid disease (CHD) is very rare manifestation of echinococcosis, representing only 2% of all celebral space occupying lesion even in the countries where the disease in is endemic. The aim of this paper is to describe the characteristic features of cerebral hydatid disease in computed tomography, magnetic resonance Imaging (MRI) and to report a multiloculated cyst (more than 100 loculi) along with its management. Case Presentation: In this paper we have reported a young boy of 20yrs with primary CHD without associated extracranial lesions with focal neurological deficits and intracranial hypertension. The extracranial investigations were found to be negative. CT and MRI of the patient suggested it to be a multiloculated Hydatid cyst. The patient was managed surgically and more than 100 daughter cysts were recovered, antihelminthic medications were given to the patient was discharged successfully. Conclusion: Multilocular Hydatid cyst is a rare SOL of Brain. When present the patient remains asymptomatic for long followed mostly by symptoms of headache and vomiting. Patients may also present with focal deficit or seizures‑Surgery remains the mainstay of treatment with careful evacuation of the cysts alongwith the medications for the causative agent (Echinococcus granulosus or multilocularies). Prevention of the disease should be given utmost emphasis.

Non-small cell lung cancer presenting with choroidal metastasis as first sign and showing good response to chemotherapy alone: a case report.

INTRODUCTION: Metastatic tumors are the most common intra-ocular malignancies and choroid is by far the most common site for intra-ocular malignancies. Multiple foci are usually involved, and bilateral involvement is frequently seen. The primary sites for choroidal metastasis in decreasing order and by gender are: breast, lung, unknown primary, gastrointestinal and pancreas, skin melanoma and other rare sources in females, and lung, unknown primary, gastrointestinal and pancreas, prostate, kidney, skin melanoma and other rare sources in males. Available treatment options are external beam radiotherapy and plaque radiotherapy, while new methods like surgical resection, transpupillary thermotherapy and intravitreal chemotherapy offer promises for the future. The use of chemotherapy alone for choroidal metastases is not widely reported. CASE PRESENTATION: We report the case of a 50-year-old Indian man who had a unilateral solitary lesion in his right eye. He was found to have an adenocarcinoma of the lung with choroidal metastasis as the first presenting sign. There were no findings of metastasis involving his contralateral eye. He was administered chemotherapy based on gemcitabine and carboplatin. He had significant progressive subjective and objective improvement since his first chemotherapy. His current best corrected visual acuity is 20/60 after three cycles of chemotherapy. CONCLUSIONS: Chemotherapy alone can be used as an effective mode of treatment in patients who have primary tumors that respond to chemotherapy.

Brain metastasis from esophageal carcinoma.

Esophageal carcinoma rarely metastasizes to brain. In our center, among 504 cases of esophageal cancer registered for treatment during a 15-year (1990-2005) period, brain metastasis from esophageal carcinoma was detected in only 1 case. An unusual case of esophageal carcinoma that presented with brain metastasis is reported here.