Biofilm exopolysaccharides alter sensory-neuron-mediated sickness during lung infection.

Infections of the lung cause observable sickness thought to be secondary to inflammation. Signs of sickness are crucial to alert others via behavioral-immune responses to limit contact with contagious individuals. Gram-negative bacteria produce exopolysaccharide (EPS) that provides microbial protection; however, the impact of EPS on sickness remains uncertain. Using genome-engineered Pseudomonas aeruginosa (P. aeruginosa) strains, we compared EPS-producers versus non-producers and a virulent Escherichia coli (E. coli) lung infection model in male and female mice. EPS-negative P. aeruginosa and virulent E. coli infection caused severe sickness, behavioral alterations, inflammation, and hypothermia mediated by TLR4 detection of the exposed lipopolysaccharide (LPS) in lung TRPV1+ sensory neurons. However, inflammation did not account for sickness. Stimulation of lung nociceptors induced acute stress responses in the paraventricular hypothalamic nuclei by activating corticotropin-releasing hormone neurons responsible for sickness behavior and hypothermia. Thus, EPS-producing biofilm pathogens evade initiating a lung-brain sensory neuronal response that results in sickness.

Translin facilitates RNA polymerase II dissociation and suppresses genome instability during RNase H2- and Dicer-deficiency.

The conserved nucleic acid binding protein Translin contributes to numerous facets of mammalian biology and genetic diseases. It was first identified as a binder of cancer-associated chromosomal translocation breakpoint junctions leading to the suggestion that it was involved in genetic recombination. With a paralogous partner protein, Trax, Translin has subsequently been found to form a hetero-octomeric RNase complex that drives some of its functions, including passenger strand removal in RNA interference (RNAi). The Translin-Trax complex also degrades the precursors to tumour suppressing microRNAs in cancers deficient for the RNase III Dicer. This oncogenic activity has resulted in the Translin-Trax complex being explored as a therapeutic target. Additionally, Translin and Trax have been implicated in a wider range of biological functions ranging from sleep regulation to telomere transcript control. Here we reveal a Trax- and RNAi-independent function for Translin in dissociating RNA polymerase II from its genomic template, with loss of Translin function resulting in increased transcription-associated recombination and elevated genome instability. This provides genetic insight into the longstanding question of how Translin might influence chromosomal rearrangements in human genetic diseases and provides important functional understanding of an oncological therapeutic target.

A comprehensive investigation of the Sr3AsX3 (X = F/Cl/Br) inorganic cubic perovskites' strain-induced structural, electronic, optical, and mechanical properties with solar cell applications.

In comparison to the lead halide perovskites, nowadays, lead-free halide perovskites have demonstrated a number of benefits, including efficient optical absorption, increased stability, variable bandgap, excellent mobility of carriers, non-toxicity, abundant raw ingredients, and low manufacturing cost. The use of the Perdew-Burke-Ernzerhof (PBE) and Heyd-Scuseria-Ernzerhof (HSE) hybrid functional inside the quantum espresso software allowed for a thorough examination of these materials, potentially leading to improvements in the development of ecologically acceptable and economically sustainable perovskite-based products. This work has extensively examined the effects of compressive and tensile strain on the structural, optical, and electronic characteristics of the inorganic cubic perovskite Sr3AsX3 (X = F, Cl, Br) with varying X anion using first-principles density-functional theory (FP-DFT). At the Γ point, the unstrained Sr3AsF3, Sr3AsCl3, and Sr3AsBr3 compounds have a direct bandgap of 1.68/2.50 eV, 1.65/2.47 eV, and 1.522/2.30 eV, respectively, from the PBE/HSE methods. A drop in bandgap values occurs when the X-anion switches from F to Cl to Br. Furthermore, the bandgaps of the three proposed structures show a minor increase in response to tensile strain and a decreasing prevalence in response to compressive strain. The optical properties, which include dielectric functions, absorption coefficient, and electron loss function, are consistent with the band characteristics of these components, all of which point to a significant capability for absorption in the visible region. The dielectric constants of Sr3AsF3, Sr3AsCl3, and Sr3AsBr3 are discovered to have peaks that, with compressive strain, redshift (move towards lower photon energy) and, under tensile strain, blueshift (move towards upper photon energy). In comparison to the compounds Sr3AsF3 and Sr3AsCl3, the parameters indicate that the material Sr3AsBr3 is more optically advantageous. The SCAPS-1D simulator was used to methodically examine the photovoltaic (PV) performance of novel cell topologies that included SnS2 as an electron transport layer (ETL) and Sr3AsF3, Sr3AsCl3, and Sr3AsBr3 as absorbers and primarily 19.76, 19.89, and 20.89% PCE was achieved, respectively.

Dietary inflammatory potential and severe headache or migraine: a systematic review of observational studies.

OBJECTIVES: We conducted the current systematic review to investigate the association between dietary inflammatory index (DII) and severe headaches or migraine among adults via synthesizing observational evidence. METHOD: We conducted a systematic literature search of observational studies through PubMed, Scopus, and Web of Science databases from inception until July 2024. The PECO framework was implemented to select eligible studies as follows: Population (adults with severe headache or migraine), Exposure (individuals with the highest adherence to a pro-inflammatory diet), Comparison (individuals with the lowest adherence to a pro-inflammatory diet), Outcome (risk of developing severe headache or migraine, headaches frequency, duration, severity, and migraine-related disability). RESULTS: After reviewing six studies involving 31,958 individuals, we found that following an anti-inflammatory diet is associated with a lower frequency and severity of migraine headaches. Additionally, our research revealed that individuals with migraines tend to have lower adherence to an anti-inflammatory diet when compared to people without migraines. Surprisingly, adherence to a pro-inflammatory diet was linked to a reduced risk of chronic daily headaches. CONCLUSION: Present findings imply a negative link between an inflammatory diet and severe headaches or migraine. However, further well-designed longitudinal studies are needed to interpret the causality and shed light on the underlying mechanisms.

Effect of thyme, ginger, and their nano-particles on growth performance, carcass characteristics, meat quality and intestinal bacteriology of broiler chickens.

This study was conducted to evaluate the effects of thyme, ginger, and their nano-particles, as alternatives to antibiotic growth promotors (AGP), on productive performance, carcass traits, meat quality and gut health of broiler chickens. A total of 270 one-day-old broiler chicks were randomly distributed into 6 groups, each consisting of 3 replicates (n = 15 chicks/replicate). The birds in group 1 were fed the control diet which contained neither antibiotic growth promotors nor phytogenic feed additives (PFA). Birds in group 2 were fed diets containing 0.05% of AGP (Bacitracin methylene disalicylate). Chicks in group 3 and 4 were fed diets supplemented with 1.0% of thyme and ginger, respectively, whereas birds in group 5 and 6 were offered diets including 0.10% of nano-thyme and nano-ginger, respectively. The experiment lasted for 35 days. It was found that thyme and ginger with their nano-products, like the antibiotic, improved the body weight, weight gain and feed conversion rate of birds. The effect of ginger and nano-ginger on body weight and weight gain was greater than other treatments. During the overall feeding period, the feed cost of production was the highest in antibiotic group, but was the lowest in ginger and nano-ginger treatments. There was no effect of dietary treatments on carcass yield or organs weight except bursa of Fabricius and abdominal fat. Thyme, ginger and their nano-composites increased the weight of bursa and reduced the abdominal fat amount. The phytogenic additives and their nano-particles improved the colour, water holding capacity, and flavor of meat. Moreover, these additives reduced the total intestinal bacterial count as well as the total aerobic mesophilic count of meat. The effect of PFA and their nano-particles on the bacterial count was similar to that of antibiotic. In conclusion, thyme and ginger with their nano- particles can be considered as promising agents in feeding of broilers to improve the growth performance, gut health and meat quality. Moreover, these additives can be used as alternatives to AGP to overcome its health hazards and the high cost. The nanotechnology of herbal plants enables them to be added in smaller amounts in poultry diets with producing the same effect of raw ingredients, and this could be due to the higher bioavailability.

Plasma activated water effects on behavior, performance, carcass quality, biochemical changes, and histopathological alterations in quail.

BACKGROUND: Plasma-activated water (PAW) is an innovative promising technology which could be applied to improve poultry health. The current study investigated the effects of drinking water supply with PAW on quail behaviour, performance, biochemical parameters, carcass quality, intestinal microbial populations, and internal organs histopathology. A total of 54 twenty-one-day-old Japanese quail chicks were randomly allotted to three treatments provided with PAW at doses 0, 1 ml (PAW-1), and 2 ml (PAW-2) per one litter drinking water. Each treatment contained 6 replicates (3 birds/ cage; one male and two females). RESULTS: The results clarified that there were no significant (P > 0.05) changes in behaviour, and performance. For the biochemical indicators, the PAW-1 group showed significantly higher serum H2O2, total protein and globulin levels compared with the other groups (P = 0.015, < 0.001, and 0.019; respectively). PAW groups had significantly lower serum creatinine and urea levels than the control (P = 0.003). For the carcass quality, the internal organs relative weight between different treatments was not changed. In contrast, there was a significant increase in the meat colour, taste, and overall acceptance scores in PAW groups compared with the control one (P = 0.013, 0.001, and < 0.001; respectively). For the intestinal microbial population, lactobacilli count was significantly higher in PAW-2 compared with the control group (P = 0.014), while there were no changes in the total bacterial count between different treatment groups. Moreover, mild histological changes were recorded in the intestine, liver, and spleen of PAW groups especially PAW-2 compared with the control one. CONCLUSIONS: PAW offered benefits, such as reducing creatine and urea levels, improving meat characteristics, and increasing lactobacilli count, all of which are crucial for sustainable quail farming. Therefore, further research is needed.

Hyaluronic acid impacts hematological endpoints and spleen histological features in African catfish (Clarias gariepinus).

Since its identification in the vitreous humour of the eye and laboratory biosynthesis, hyaluronic acid (HA) has been a vital component in several pharmaceutical, nutritional, medicinal, and cosmetic uses. However, little is known about its potential toxicological impacts on aquatic inhabitants. Herein, we investigated the hematological response of Clarias gariepinus to nominal doses of HA. To achieve this objective, 72 adult fish were randomly and evenly distributed into four groups: control, low-dose (0.5 mg/l HA), medium-dose (10 mg/l HA), and high-dose (100 mg/l HA) groups for two weeks each during both the exposure and recovery periods. The findings confirmed presence of anemia, neutrophilia, leucopoenia, lymphopenia, and eosinophilia at the end of exposure to HA. In addition, poikilocytosis and a variety of cytomorphological disturbances were observed. Dose-dependent histological alterations in spleen morphology were observed in the exposed groups. After HA removal from the aquarium for 2 weeks, the groups exposed to the two highest doses still exhibited a notable decline in red blood cell count, hemoglobin concentration, mean corpuscular hemoglobin concentration, and an increase in mean corpuscular volume. Additionally, there was a significant rise in neutrophils, eosinophils, cell alterations, and nuclear abnormalities percentages, along with a decrease in monocytes, coupled with a dose-dependent decrease in lymphocytes. Furthermore, only the highest dose of HA in the recovered groups continued to cause a significant increase in white blood cells. White blood cells remained lower, and the proportion of apoptotic RBCs remained higher in the high-dose group. The persistence of most of the haematological and histological disorders even after recovery period indicates a failure of physiological compensatory mechanisms to overcome the HA-associated problems or insufficient duration of recovery. Thus, these findings encourage the inclusion of this new hazardous agent in the biomonitoring program and provide a specific pattern of hematological profile in HA-challenged fish. Further experiments are highly warranted to explore other toxicological hazards of HA using dose/time window protocols.

Application of gelatin-based zinc oxide nanoparticles bionanocomposite coatings to control Listeria monocytogenes in Talaga cheese and camel meat during refrigerated storage.

Listeria monocytogenes is a concerning foodborne pathogen incriminated in soft cheese and meat-related outbreaks, highlighting the significance of applying alternative techniques to control its growth in food. In the current study, eco-friendly zinc oxide nanoparticles (ZnO-NPs) were synthesized using Rosmarinus officinalis, Punica granatum, and Origanum marjoram extracts individually. The antimicrobial efficacy of the prepared ZnO-NPs against L. monocytogenes was assessed using the agar well diffusion technique. Data indicated that ZnO-NPs prepared using Origanum marjoram were the most effective; therefore, they were used for the preparation of gelatin-based bionanocomposite coatings. Furthermore, the antimicrobial efficacy of the prepared gelatin-based bionanocomposite coatings containing eco-friendly ZnO-NPs was evaluated against L. monocytogenes in Talaga cheese (an Egyptian soft cheese) and camel meat during refrigerated storage at 4 ± 1 oC. Talaga cheese and camel meat were inoculated with L. monocytogenes, then coated with gelatin (G), gelatin with ZnO-NPs 1% (G/ZnO-NPs 1%), and gelatin with ZnO-NPs 2% (G/ZnO-NPs 2%). Microbiological examination showed that the G/ZnO-NPs 2% coating reduced L. monocytogenes count in the coated Talaga cheese and camel meat by 2.76 ± 0.19 and 2.36 ± 0.51 log CFU/g, respectively, by the end of the storage period. Moreover, G/ZnO-NPs coatings controlled pH changes, reduced water losses, and improved the sensory characteristics of Talaga cheese and camel meat, thereby extending their shelf life. The obtained results from this study indicate that the application of gelatin/ZnO-NPs 2% bionanocomposite coating could be used in the food industry to control L. monocytogenes growth, improve quality, and extend the shelf life of Talaga cheese and camel meat.

Deciphering the Mechanisms and Biotechnological Implications of Nanoparticle Synthesis Through Microbial Consortia.

Nanomaterial synthesis is a growing study area because of its extensive range of uses. Nanoparticles' high surface-to-volume ratio and rapid interaction with various particles make them appealing for diverse applications. Traditional physical and chemical methods for creating metal nanoparticles are becoming outdated because they involve complex manufacturing processes, high energy consumption, and the formation of harmful by-products that pose major dangers to human health and the environment. Therefore, there is an increasing need to find alternative, cost-effective, dependable, biocompatible, and environmentally acceptable ways of producing nanoparticles. The process of synthesizing nanoparticles using microbes has become highly intriguing because of their ability to create nanoparticles of varying sizes, shapes, and compositions, each with unique physicochemical properties. Microbes are commonly used in nanoparticle production because they are easy to work with, can use low-cost materials, such as agricultural waste, are cheap to scale up, and can adsorb and reduce metal ions into nanoparticles through metabolic activities. Biogenic synthesis of nanoparticles provides a clean, nontoxic, ecologically friendly, and sustainable method using renewable ingredients for reducing metals and stabilizing nanoparticles. Nanomaterials produced by bacteria can serve as an effective pollution control method due to their many functional groups that can effectively target contaminants for efficient bioremediation, aiding in environmental cleanup. At the end of the paper, we will discuss the obstacles that hinder the use of biosynthesized nanoparticles and microbial-based nanoparticles. The paper aims to explore the sustainability of microorganisms in the burgeoning field of green nanotechnology.

Effectiveness of ChatGPT in remote learning environments: An empirical study with medical students in Saudi Arabia.

Purpose: This study aims to assess the effectiveness of ChatGPT in remote learning among medical students. Methods: This cross-sectional survey study recruited 386 medical students from three public universities in Saudi Arabia. Participants completed an online questionnaire designed to assess perceptions of ChatGPT's effectiveness in remote learning. The questionnaire included Likert scale questions to evaluate various aspects of ChatGPT's support in remote learning, such as personalized learning, language and communication skills, and interactive quizzing. Data were analyzed using SPSS, employing descriptive statistics, independent samples t-tests, one-way ANOVA, and Cronbach's alpha to evaluate reliability. Results: Participants mostly used ChatGPT on a weekly (43.2%) or daily (48.7%) basis, primarily on personal computers (62.5%). Mean scores for ChatGPT's support in remote learning were high for personalized learning (4.35), language and communication skills (4.23), and interactive quizzing and assessments (4.01). Statistically significant differences were found based on gender for interactive quizzing (p = .0177) and continuity of education (p = .0122). Conclusion: Despite certain challenges and variations in perceptions based on gender and education level, the overwhelmingly positive attitudes toward ChatGPT highlight its potential as a valuable tool in medical education.

Enhancing medical students critical thinking skills through ChatGPT: An empirical study with medical students.

STUDY PURPOSE: This study aims to assess the effectiveness of ChatGPT in critical thinking skills among medical students. METHODS: This cross-sectional survey study recruited 392 medical students from three public universities in Saudi Arabia. Participants completed an online questionnaire assessing perceptions of ChatGPT's impact on critical thinking skills. Data were analyzed using SPSS, employing descriptive statistics, t-tests, analysis of variance, and Cronbach's alpha to evaluate reliability. RESULTS: Significant gender-based differences were found in perceptions of ChatGPT's efficacy, particularly in generating diverse perspectives (P = 0.0407*) and encouraging questioning (P = 0.0277*). Reflective practice perceptions varied significantly by age (P = 0.0302*), while academic backgrounds yielded significant differences across all factors assessed (P < 0.0001*). Overall, 92.6% believed integrating ChatGPT would benefit critical thinking skills. Most participants (N = 174) strongly agreed that ChatGPT improved critical thinking. CONCLUSION: Integrating ChatGPT into medical education could offer valuable opportunities for fostering critical thinking abilities, albeit with the need for addressing associated challenges and ensuring inclusivity.

Role of Eruca sativa L. seeds in boosting the reproductive performance of male Japanese quails (Coturnix c. japonica).

Herein we attempt to shed light on the potential improving effect of Eruca sativa seeds (ESS) on the reproductive aspects of male Japanese quails. To accomplish this objective, two groups of quails were supplemented with ESS powder at doses of 5 and 10 g/kg feed from 7 days to 140 days of age, in addition to the control group, which did not receive treatment. Forty males were reared singly in cages to evaluate sperm characters and 32 males were raised with 64 females to evaluate fertility and sperm penetrability. Sixty-six phytochemical compounds were found according to gas chromatography-mass spectrometry analysis of ESS. The most plentiful ones are 13-docosenoic acid methyl ester, 9-octadecenoic acid methyl ester, and linoleic acid methyl ester. Both 5 g/kg and 10 g/kg doses of ESS showed similar effectiveness in enhancing various reproductive parameters, including gonadal index, sperm characteristics, fertility, libido, and cloacal gland attributes. However, some aspects like sperm concentration and testosterone levels exhibited a dose-dependent response. There is no significant change in mortality rate of supplemented groups compared to the control one. ESS also caused a reduction in feed intake and an enhancement in feed conversion ratio without affecting final body weight and body weight gain. This suggests potential nutritional benefits beyond reproductive health. The low-dose-fed group showed a significant reduction in total cholesterol and malondialdehyde compared to the high-dose-fed and unfed groups. The higher dose notably increased total antioxidant capacity compared to the lower dose and control group. Despite the positive effects on male reproductive parameters, there wasn't a significant impact on hatchability percentage, indicating that while male fertility improved, it might not have directly affected the viability of the eggs. Overall, the study suggests that ESS could be a safe and promising addition to the diet of male Japanese quails to enhance their reproductive capabilities without adverse effects. The findings could have implications for poultry farming by potentially improving breeding efficiency and health outcomes in quails.

Pyrimidine-morpholine hybrids as potent druggable therapeutics for Alzheimer's disease: Synthesis, biochemical and in silico analyses.

The identification of effective and druggable cholinesterase inhibitors to treat progressive neurodegenerative Alzheimer's disorder remains a continuous drug discovery hunt. In this perspective, the present study investigates the design and discovery of pyrimidine-morpholine hybrids (5a-l) as potent cholinesterase inhibitors. Palladium-catalyzed Suzuki-Miyaura cross-coupling reaction was employed to introduce the structural diversity on the pyrimidine heterocyclic core. A range of commercially available boronic acids was successfully coupled showing a high functional group tolerance. In vitro cholinesterase inhibitory potential using Ellman's method revealed significantly strong potency. Compound 5h bearing a meta-tolyl substituent at 2-position of pyrimidine ring emerged as a lead candidate against AChE with an inhibitory potency of 0.43 ± 0.42 µM, ∼38-fold stronger value than neostigmine (IC50 = 16.3 ± 1.12 µM). Compound 5h also showed the lead inhibition against BuChE with an IC50 value of 2.5 ± 0.04 µM. The kinetics analysis of 5h revealed the non-competitive mode of inhibition against AChE whereas computational modelling results of potent leads depicted diverse contacts with the binding site amino acid residues. Molecular dynamics simulations revealed the stability of biomolecular system, while, ADME analysis demonstrated druglikeness behaviour of potent compounds. Overall, the investigated pyrimidine-morpholine scaffold presented a remarkable potential to be developed as efficacious anti-Alzheimer's drugs.

Development of pyrolo[2,3-c]pyrazole, pyrolo[2,3-d]pyrimidine and their bioisosteres as novel CDK2 inhibitors with potent in vitro apoptotic anti-proliferative activity: Synthesis, biological evaluation and molecular dynamics investigations.

Inhibiting the CDK2/cyclin A2 enzyme has been validated in multiple clinical manifestations related to multiple types of cancer. Herein, novel series of pyrolo[2,3-c]pyrazole, pyrolo[2,3-c]isoaxazole and pyrolo[2,3-d]pyrimidine, pyrolo[3,2-c]pyridine & indole based analogs were designed, synthesized and biologically evaluated for their in vitro antiproliferative activity where the obtained results revealed that most of the newly synthesized compounds showed significant cytotoxic activity towards MCF-7 (breast cancer cell lines) and HepG-2 (hepatocellular carcinoma) with IC50 ranging from 3.20 µM to 10.05 µM & from 2.18 µM to 13.49 µM, respectively, compared to that of Sorafenib (IC50 9.76 & 13.19 µM, respectively). The in vitro inhibitory profile of the most promising compounds (9, 11, 14, 15, 16, 17 and 20) towards CDK2/CyclinA2 was evaluated. Compounds 14 & 15 exhibited potent inhibitory profile against CDK2 with (IC50 0.11 and 0.262 µM, respectively comparable to Sorafenib IC50 0.184 µM. Western blotting of 14 & 15 at MCF-7 cell line confirmed the diminishing activity on CDK2. Furthermore, both compounds exserted a significant cell cycle arrest and apoptosis. Moreover, the normal cell line cytotoxicity for both compounds revealed low cytotoxic results in normal cells rather than cancer cells. Molecular docking and dynamic simulation validated the potentiality of the newly synthesized compounds to have high binding affinity within CDK2 binding pocket. 3DQSAR pharmacophore, in-silico ADME/TOPKAT studies and drug-likeness showed proper pharmacokinetic properties and helped in structure requirements prediction. The obtained model and pattern of substitution could be used for further development of CDK2 inhibitors.

The effects of Ginger (Zingiber officinale) roots on the reproductive aspects in male Japanese Quails (Coturnix coturnix japonica).

BACKGROUND: The Japanese quail is considered one of the most significant species in the poultry industry. However, the high male-to-female ratio results in the aggressive behavior of males. Dietary strategies that improve the properties of semen could reduce the number of males required to maintain optimal fertility and reduce aggressive behavior. Therefore, this study aims to provide insight into the possible improving efm fect of ginger roots on the reproductive aspects of Japanese male quails. RESULTS: To achieve this objective, powder of Ginger roots was administrated to 2 groups of quails (10, and 15 g/Kg feed) from 7 days until 70 days of age. Some males were reared singly in cages (n = 40 for each group) to assess sperm quality and other males (n = 32 for each group) were raised with females to assess fertility and sperm-egg penetration. Additionally, biochemical tests and histological examination were also performed. When compared to the control group, dietary inclusion of Ginger at a dose of 15 g caused more improvement in ejaculate volume, sperm concentration, motility, viability and sperm-egg penetration. Whereas, the motility and fertility percentages of sperms were equipotent in both doses. Dose-dependent increases were found in the cloacal gland area and volume, as well as foam production and weight. Both doses resulted in a significant reduction in plasma total cholesterol along with an elevation cin plasma testosterone and lipid peroxides. The comparison between all groups concerning nitric oxide, catalase, superoxide dismutase, and total antioxidant capacity revealed the absence of significant difference. Morphologically, the diameter of the seminiferous tubules and the height of germinal epithelium significantly increased especially in the higher dose of Ginger. CONCLUSIONS: Ginger roots especially at a dose of 15 gm/kg feed was effective in improving male reproductive performance. These findings are of utmost importance in encouraging the addition of Ginger roots in ration formulation in male quails.

A disorder clinically resembling cystic fibrosis caused by biallelic variants in the AGR2 gene.

PURPOSE: We sought to describe a disorder clinically mimicking cystic fibrosis (CF) and to elucidate its genetic cause. METHODS: Exome/genome sequencing and human phenotype ontology data of nearly 40 000 patients from our Bio/Databank were analysed. RNA sequencing of samples from the nasal mucosa from patients, carriers and controls followed by transcriptome analysis was performed. RESULTS: We identified 13 patients from 9 families with a CF-like phenotype consisting of recurrent lower respiratory infections (13/13), failure to thrive (13/13) and chronic diarrhoea (8/13), with high morbidity and mortality. All patients had biallelic variants in AGR2, (1) two splice-site variants, (2) gene deletion and (3) three missense variants. We confirmed aberrant AGR2 transcripts caused by an intronic variant and complete absence of AGR2 transcripts caused by the large gene deletion, resulting in loss of function (LoF). Furthermore, transcriptome analysis identified significant downregulation of components of the mucociliary machinery (intraciliary transport, cilium organisation), as well as upregulation of immune processes. CONCLUSION: We describe a previously unrecognised autosomal recessive disorder caused by AGR2 variants. AGR2-related disease should be considered as a differential diagnosis in patients presenting a CF-like phenotype. This has implications for the molecular diagnosis and management of these patients. AGR2 LoF is likely the disease mechanism, with consequent impairment of the mucociliary defence machinery. Future studies should aim to establish a better understanding of the disease pathophysiology and to identify potential drug targets.

Investigation of novel bis-thiadiazole bearing schiff base derivatives as effective inhibitors of thymidine phosphorylase: Synthesis, in vitro bioactivity and molecular docking study.

Thymidine phosphorylase (TP) is an angiogenic enzyme. It is crucial for the development, invasion and metastasis of tumors as well as angiogenesis. In our current research, we examine how structurally changing bis-thiadiazole bearing bis-schiff bases affects their ability to inhibit TP. Through the oxidative cyclization of pyridine-based bis-thiosemicarbazone with iodine, a series of fourteen analogs of bis-thiadiazole-based bis-imines with pyridine moiety were developed. Newly synthesized scaffolds were assessed in vitro for their thymidine phosphorylase inhibitory potential and showed moderate to good inhibition profile. Eleven scaffolds such as 4a-4d,4f-4 h and 4j-4 m were discovered to be more effective than standard drug at inhibiting the thymidine phosphorylase enzyme with IC50 values of 1.16 ± 1.20, 1.77 ± 1.10, 2.48 ± 1.30, 12.54 ± 1.60, 14.63 ± 1.70, 15.53 ± 1.80, 17.47 ± 1.70, 18.98 ± 1.70, 19.53 ± 1.50, 22.73 ± 2.40 and 24.87 ± 2.80 respectively, while remaining three analogs such as 4n, 4i and 4ewere found to be more potent, but they were less potent than the standard drug. All analogs underwent SAR studies based on the pattern of substitutions around the aryl part of the bis-thiadiazole skeleton. The most active analogs in the synthesized series were then molecular docking study performed to investigate their interactions of active part of enzyme. The results showed that remarkable interactions were exhibited by these analogs with the targeted enzymes active sites. Furthermore, to confirm the structure of synthesized analogs by employing spectroscopic tools such as HREI-MS and NMR.

Synthesis, carbonic anhydrase enzyme inhibition evaluations, and anticancer studies of sulfonamide based thiadiazole derivatives.

The sulfonamide-based thiadiazole derivatives (STDs) with different hydrophobic/hydrophilic substitutions were synthesized to investigate their potentials in carbonic anhydrase inhibition (CAI). The CAI activity of the STDs (4a-4h) and the mechanism of the inhibition kinetics were determined. STD 4f contained both methoxy and Cl groups at benzene ring in STD 4f showed the lowest IC50 value. The molecular docking study confirmed that STDs bind strongly with the active sites of the target protein PDBID 1V9E. With the help of Lineweaver-Burk plots, inhibition kinetics of PDBIR 1V9E protein with STDs were determined. Cytotoxicity was checked against human keratinocyte cell lines and the anticancer properties were determined against MCF-7 cell lines. The electrochemical method was used to investigate the binding study with DNA and CA enzymes. Anticancer studies showed that STDs have weak bonding ability to DNA and strong binding ability with CA. It is concluded that anticancer activity is through CAI rather than by DNA binding.

Synthesis, biological evaluation, and in silico studies of new CDK2 inhibitors based on pyrazolo[3,4-d]pyrimidine and pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine scaffold with apoptotic activity.

Cyclin-dependent kinase inhibition is considered a promising target for cancer treatment for its crucial role in cell cycle regulation. Pyrazolo pyrimidine derivatives were well established for their antitumor activity via CDK2 inhibition. In this research, new series of pyrazolopyrimidine derivatives (4-15) was designed and synthesised as novel CDK2 inhibitors. The anti-proliferative activities against MCF-7, HCT-116, and HepG-2 were used to evaluate their anticancer activity as novel CDK2 inhibitors. Most of the compounds showed superior cytotoxic activity against MCF-7 and HCT-116 compared to Sorafenib. Only compounds 8, 14, and 15 showed potent activity against HepG-2. The CDK2/cyclin A2 enzyme inhibitory activity was tested for all synthesised compounds. Compound 15 showed the most significant inhibitory activity with IC50 0.061 ± 0.003 µM. It exerted remarkable alteration in Pre G1 and S phase cell cycle progression and caused apoptosis in HCT cells. In addition, the normal cell line cytotoxicity for compound 15 was assigned revealing low cytotoxic results in normal cells rather than cancer cells. Molecular docking was achieved on the designed compounds and confirmed the two essential hydrogen binding with Leu83 in CDK2 active site. In silico ADMET studies and drug-likeness showed proper pharmacokinetic properties which helped in structure requirements prediction for the observed antitumor activity.

Marine Alga Ulva fasciata-Derived Molecules for the Potential Treatment of SARS-CoV-2: An In Silico Approach.

SARS-CoV-2 is the causative agent of the COVID-19 pandemic. This in silico study aimed to elucidate therapeutic efficacies against SARS-CoV-2 of phyco-compounds from the seaweed, Ulva fasciata. Twelve phyco-compounds were isolated and toxicity was analyzed by VEGA QSAR. Five compounds were found to be nonmutagenic, noncarcinogenic and nontoxic. Moreover, antiviral activity was evaluated by PASS. Binding affinities of five of these therapeutic compounds were predicted to possess probable biological activity. Fifteen SARS-CoV-2 target proteins were analyzed by the AutoDock Vina program for molecular docking binding energy analysis and the 6Y84 protein was determined to possess optimal binding affinities. The Desmond program from Schrödinger's suite was used to study high performance molecular dynamic simulation properties for 3,7,11,15-Tetramethyl-2-hexadecen-1-ol-6Y84 for better drug evaluation. The ligand with 6Y84 had stronger binding affinities (-5.9 kcal/mol) over two standard drugs, Chloroquine (-5.6 kcal/mol) and Interferon α-2b (-3.8 kcal/mol). Swiss ADME calculated physicochemical/lipophilicity/water solubility/pharmacokinetic properties for 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, showing that this therapeutic agent may be effective against SARS-CoV-2.