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The rapid evolution of CRISPR technology has revolutionized molecular biology, and among the various systems, CRISPR/Cas12a stands out for its high specificity and efficient collateral cleavage activity. This review article focuses on the recent advancements and applications of split technology within the CRISPR/Cas12a framework, highlighting its transformative role in molecular diagnostics and biosensing. Split technology innovatively divides functional nucleic acid components into modular segments that are activated by specific targets, significantly enhancing the specificity and sensitivity of biosensors. This design addresses the inherent limitations of traditional sensor systems, enabling the direct detection of ultrashort nucleic acids and improved discrimination of single-nucleotide variants, thereby facilitating the simultaneous detection of multiple biomolecules. The versatility of split-enabled biosensors extends beyond genetic testing, making them valuable tools in diagnostics, therapeutics, and environmental science. Despite challenges such as crRNA degradation and reassembly kinetics, ongoing research and engineering solutions continue to enhance the stability and performance of these systems. This review synthesizes the foundational principles, recent advancements, and potential applications of split technology while also identifying challenges and opportunities for future exploration. Ultimately, our insights provide a comprehensive resource to leverage the full potential of CRISPR/Cas12a-based split technology in advancing biosensing methodologies and clinical applications.

Objectives: To compare the efficacy of fluoxetine and N-acetylcysteine (NAC) in reducing craving, relapse, and improving treatment adherence in men with alcohol dependence syndrome (ADS). Methods: Men aged 18 to 65 years with a diagnosis of ADS who had maintained >=7 days of abstinence from alcohol were recruited. Participants were randomised in a 1:1 ratio to receive either oral fluoxetine 20 mg/day or NAC 600 mg twice/day for 12 weeks. Participants were followed up at weeks 4, 8, and 12. Outcome measures included the Penn Alcohol Craving Scale (PACS), relapse rate, treatment adherence, and adverse effects. Results: In total, 100 men were equally randomised to receive either fluoxetine or NAC. Both fluoxetine and NAC were effective in lowering PACS scores over 12 weeks, with scores being lower in the NAC group than in the fluoxetine group at week 4 (15.4 vs 17.6, p equivalent 0.03), week 8 (11.3 vs 14.2, p equivalent 0.002), and week 12 (7.8 vs 11.1, p < 0.001). The reduction in PACS scores was significantly greater in the NAC group from week 4 onward. Relapse rates were lower in the NAC group (18.0% vs 32.0%, p equivalent 0.049). Treatment adherence was higher (but not significantly) in the NAC group (90.0% vs 84.0%, p equivalent 0.38). Both medications were well tolerated. All adverse effects were mild. Conclusion: Both fluoxetine and NAC were effective in reducing alcohol craving over 12 weeks, with greater reduction in the NAC group at each follow-up assessment. The NAC group also had a lower relapse rate and better adherence and tolerability.

A 24 kDa leucine-rich protein from ion exchange fractions of Solanum trilobatum, which has anti-bacterial activity against both the Gram-negative Vibrio cholerae and Gram-positive Staphylococcus aureus bacteria has been purified. In this study, mass spectrometry analysis identified the leucine richness and found a luminal binding protein (LBP). Circular dichroism suggests that the protein was predominantly composed of α- helical contents of its secondary structure. Scanning electron microscopy visualized the characteristics and morphological and structural changes in LBP-treated bacterium. Further in vitro studies confirmed that mannose-, trehalose- and raffinose-treated LBP completely inhibited the hemagglutination ability towards rat red blood cells. Altogether, these studies suggest that LBP could bind to sugar moieties which are abundantly distributed on bacterial surface which are essential for maintaining the structural integrity of bacteria. Considering that Solanum triolbatum is a well-known medicinal and edible plant, in order to shed light on its ancient usage in this work, an efficient anti-microbial protein was isolated, characterized and its in vitro functional study against human pathogenic bacteria was evaluated.

This work reports the synthesis of silver nanoparticles (AgNPs) using aqueous extract of Plumbago auriculata, and evaluates their antibacterial and larvicidal activities. The synthesized silver nanoparticles were characterized by various spectroscopy techniques, such as FTIR, XRD, TEM, EDX, Zeta potential, and DLS. The synthesized AgNPs exhibited significant antibacterial activity against Gram-positive and Gram-negative bacteria, such as Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae. Furthermore, synthesized nanoparticles inhibited the fourth instars larvae of Aedes aegypti and Culex quinquefasciatus at the concentration of 45.1 and 41.1 µg/mL respectively. Results of dose-dependent studies showed that synthesized nanoparticles were also effective at low concentrations. Molecular docking studies performed with the salivary protein and odorant-binding protein of Aedes aegypti and Culex quinquefasciatus demonstrated that the naphthoquinone compound plumbagin exhibited reliable binding affinity towards the two enzymes. The findings thus reveal that the plant extract and its nanoparticles can be a better alternative to available chemicals to control mosquitos.

Background The fibula, situated laterally in the leg, receives vital nutrition through nutrient arteries during embryonic bone growth and early ossification. This study aims to assess the direction, distance, location, number, and foraminal index of nutrient foramina in dry fibulae from the South Indian population. Materials and methods A descriptive cross-sectional analysis involved 63 dry adult human fibulae sourced from the Department of Anatomy, Saveetha Medical College and Hospital, Thandalam. Parameters like fibula length, location, number, and direction of vascular foramina were recorded. Statistical analyses were performed on morphometric data and foraminal index. Results The mean fibula length was 34.68 ± 2.11 cm. Among the fibulae, 88.88% had a single nutrient foramen, 4.76% had dual foramina, and 6.34% lacked nutrient foramina. Most single foramina were found on the medial crest (66.66%), followed by between the medial crest and posterior border (20.63%). Nutrient foramina were primarily located in Zone II (87.30%), followed by Zone III (11.11%) and Zone I (1.58%). Directionally, 85.71% pointed downward, while 14.28% pointed upward. The mean foraminal index was 40.85 ± 6.78, ranging from 32.57 to 56.25. Conclusion Zone II, particularly on the medial crest, was the most prevalent location for vascular foramina in the fibula. Dual foramina occurred in 6.34% of cases. This precise anatomical knowledge is valuable for various medical professionals, including anthropologists, forensic experts, radiologists, plastic surgeons, and orthopedic surgeons, especially in procedures involving vascularized fibular bone grafts.

Background/Objectives: Phloroglucinol (PHL), a phenolic compound extracted from the brown alga Rosenvingea intricata, exhibits potent antioxidant and anticancer properties. This study aims to extract, purify, and characterize PHL, and further develop functionalized zinc oxide nanoparticles (ZnO NPs) loaded with PHL to enhance its therapeutic potential. Methods: PHL was extracted using acetone and purified through Sephadex LH-20 column chromatography, yielding a highly enriched fraction (F-3). The purified compound was characterized by FTIR, HPLC, NMR, and LC-MS. ZnO NPs were synthesized, PEGylated, and conjugated with PHL, forming ZnO-PEG-PHL NPs. Their characterization included DLS, zeta potential, XRD, SEM-EDAX, and encapsulation efficiency studies. Antioxidant assays (DPPH, FRAP, ABTS, RPA) were performed and in vitro cytotoxicity on A549 lung cancer cells were determined to evaluate the therapeutic efficacy of PHL. Results: The purified PHL fraction showed a high phenolic content (45.65 PHL mg/g), which was was confirmed by spectral analysis. The ZnO-PEG-PHL NPs increased in size from 32.36 nm to 46.68 nm, with their zeta potential shifting from −37.87 mV to −26.82 mV. The antioxidant activity was superior for the ZnO-PEG-PHL NPs in all assays, while the in vitro cytotoxicity tests showed an IC50 of 40 µg/mL compared to 60 µg/mL for the ZnO NPs and 70 µg/mL for PHL. Apoptotic studies revealed significant cell cycle arrest and apoptosis induction. Conclusions: The synthesized ZnO-PEG-PHL NPs demonstrated enhanced antioxidant and anticancer properties, making them promising candidates for cancer therapy and antioxidant applications.

Bimetallic nanocomposites represent an important class of nanomaterials with diverse applications in optoelectronics, catalysis, bioimaging, and antimicrobial therapy. In the present study, Gs@Zn/Cu nanocomposites were synthesized via a green synthesis approach. Gracilaria spinulosa seaweed extract was employed to simultaneously load zinc/copper nanoparticles, utilizing its bioactive components as reducing and binding agents. Antibacterial assays demonstrated that the synthesized nanocomposite possesses notable antibacterial activity against Escherichia coli and Staphylococcus aureus . The combination of zinc and copper produced inhibition zones of 20 mm against E. coli and 16 mm against S. aureus . Antibiofilm assays indicated that the Gs@Zn/Cu nanocomposite reduced approximately 85% of biofilm formation at the lowest tested concentrations. Artemia biosafety assays indicated that the synthesized Gs@Zn/Cu nanocomposites are non-toxic and may be suitable for clinical applications. Collectively, these results suggest that bioactive Gs@Zn/Cu nanocomposites have considerable potential as antimicrobial and biomedical materials.

The biogenic synthesis of nanomaterials has great advantages, biocompatible, and valuable material production. In this study, we demonstrated a sustainable and biocompatible approach for the synthesis of Ag nanoparticles (Ag NPs) using the endophytic fungi Neocosmospora solani (NS) extract isolated from the Anaphalis contorta stem. The anticancer and antibacterial potential of bioactive NS mediated AgNPs was evaluated by in vitro studies. The physicochemical characteristics of synthesized NS-AgNPs were systematically investigated by UV-vis, XRD, FTIR, FE-SEM, DLS and Zeta potential analyzer. In details, UV-visible spectroscopy confirmed the presence of biosynthesized NS-AgNPs at 432 nm, while XRD analysis confirmed their crystal nature. The FTIR spectrum confirmed the presence of functional groups in biomolecules that act as a capping agent for the nanoparticles. SEM was used to evaluate the shape of AgNPs. Dynamic light scattering (DLS) indicated that the average particle size is 362.3 nm, a zeta potential was − 0.168 mV with a single peak. The biosystem technique produced stable AgNPs up to 2 months following synthesis. In addition, the NS-AgNPs were exposed to excellent antibacterial efficacy against human pathogenic bacterial strains. The results of the anticancer assessment of NS-AgNPs against A549 lung cancer cells revealed that the dose-dependent cytotoxic and morphological changes have been reported in both AO/EB and Hoechst’s staining assays. In the future, it might be an excellent antibiotic and anticancer material for biomedical applications.

Medicinal plant-based biocompatible nanomaterials have become increasingly important in the healthcare field. Zeolitic imidazole (ZIF-8) frameworks are fascinating porous crystalline materials that have great attention in the field of drug delivery due to their biodegradability, pH sensitivity to changes in acidic conditions, and chemical durability. The development of ZIF-8 particles loaded with Achyranthes aspera extract (AA) and the evaluation of their anticancer properties are the primary findings of this study. Various spectroscopy and macroscopic strategies were utilized to completely investigate the material characteristics of the AA@ZIF-8 nanocomposite. MTT results of the AA@ZIF-8 nanocomposite demonstrated significant anticancer potential against A549 cells with an IC 50 value of 63.22 ± 0.03 μg/mL. Fluorescent microscopy investigations have demonstrated that enhanced levels of reactive oxygen species (ROS) can lead to DNA damage, which could be the possible cause for the cytotoxic effects of an AA@ZIF-L nanocomposite on the A549 cells. The materials’ biocompatibility has been proven via the Artemia salina acute toxicity test. The Artemia salina acute toxicity assay results confirmed that the nanocomposites’ LC 50 value was 160.13 ± 2.82 μg/mL, with a nontoxic nature. According to the research findings, AA@ZIF-8 nanocomposite is a promising biocompatible material for drug delivery system for lung cancer therapy and other biomedical applications.

Brassica oleracea Linn. a dietary rich vegetable with high potential health properties, remarkably its secondary metabolites possessing interesting pharmacological properties. In the twenty-first century, nanoparticles and nano-based biomedical applications play a dominant role in the health sector. Silver nanoparticles are one such metallic nanoparticle that has potential application in the pharma field and act as a potent source against microbial infections. The biosynthesized silver nanoparticles from Brassica oleracea were characterized using various spectroscopy and electron microscopy techniques. Among the diverse secondary metabolites found in Brassica oleracea L., the methanolic extract of the leaf showed the presence of different phytochemical analysis present in the plant sample. Phenolic compounds and flavonoids have received considerable attention for their potent anti-oxidative properties. Based on the quantitative analysis of phytochemical studies, the phenols and flavonoids were found to be in desirable concentrations. Phyto-synthesized silver nanoparticles were visually validated by their color change and confirmed with the spectral analysis by the presence of peak values that matched with the synthesized level of silver nanoparticles. Brassica oleracea leaf extract was screened for its anti-bacterial efficacy and evaluated against Gram-positive and Gram-negative strains using the disc diffusion method. As a result, the phyto-synthesized silver nanoparticles showed promising results against pathogenic bacterial strains.