A longstanding challenge in surfactant assembly is accessing vesicle and condensed states in low-molecular-weight systems without precipitation or severe kinetic trapping while obtaining structural insights into the resulting mesostructures. Here, we show that a two-component mixture of decylammonium bromide (DeAB) and sodium salicylate (NaSal) exhibits concentration-dependent vesicle and coacervate formation at low surfactant concentrations. At a fixed [DeAB] = 10 mM, well below the critical micelle concentration, equimolar NaSal (10 mM) lowers the aggregation threshold and produces precipitation-free vesicles, consistent with a multilamellar organization. By contrast, increasing [NaSal] to 50 mM induced liquid-liquid phase separation, forming an amphiphile-rich condensed phase. Freeze-fracture transmission electron microscopy (TEM) and two- and three-dimensional cryo-TEM further revealed that the condensed phase was structurally heterogeneous, containing persistent internal voids and coexisting nanoscale architectures. Together, these results identify the DeAB/NaSal system as a molecularly simple platform in which vesicle and coacervate states can be accessed through counterion concentration. This system provides an experimentally tractable basis for mechanistic studies of structured surfactant condensates, including protocell-relevant models.
Understanding how functionalized nanoparticles interact with double-stranded DNA at the interface is essential for two purposes. It helps interpret nanoparticle-induced nucleic acid deformation and guides the design of DNA-active nanomaterials. Here, we use all-atom molecular dynamics simulations, umbrella sampling, and electrostatic/contact analyses to examine the sequence and DNA-size dependence of the interactions between a cysteine-functionalized CdTe nanoparticle and dsDNA, with groove geometry serving as a key interfacial binding mode. For both 32-bp DNA sequences studied, nanoparticle binding is attractive in both groove orientations, but the major-groove pathway is consistently more favorable than the minor-groove pathway. The free-energy minimum for the GATATC-containing DNA is deeper than that for the ATCGAT mutant, especially in the major groove, indicating clear sequence dependence in groove-mediated interfacial binding. Structural analysis further shows that stronger major-groove binding is associated with more extensive nanoparticle-DNA contacts and a more favorable local electrostatic environment. Nanoparticle adsorption induces measurable DNA bending. The bending response follows the same trend as the interfacial binding strength. Specifically, major-groove binding produces stronger bending than minor-groove binding. The GATATC-containing DNA bends more strongly than the ATCGAT mutant. The bending response also weakens with increasing DNA size, as the duplex length increases from 32 bp to 80-90 bp. Analyses of base stacking and phosphodiester-bond fluctuations further indicate that local structural stability differs between A-T and G-C base pairs. These local differences contribute to the observed DNA bending. These results define the CdTe nanoparticle-DNA interaction as a sequence- and size-dependent interfacial recognition process that is mediated by groove-dependent binding, in which interfacial contacts, electrostatic interactions, sequence, and DNA length jointly govern binding free energy and DNA deformation.
This manuscript investigates the limitations of enzyme-electrocatalyst coupling in one-step designed amperometric cholesterol biosensors. One-step electrodeposition from Pd-ChOx-Nafion electrolyte produces a bioinorganic hybrid sensing layer, while preserving the enzyme's native conformation and biocatalytic activity. Despite structural coupling between Pd and ChOx, the amperometric response remains dominated by enzymatic H2O2 production, indicating that direct electronic communication between ChOx and the electrode is not established. More specifically, we show that direct electron transfer (DET) cannot occur even in ultrathin electrodeposited nanostructured films, likely due to oxygen-dominated electrochemistry. Furthermore, the sensitivity of cholesterol determination using Pd-NPs/ChOx/Nafion-modified electrodes was independent of layer thickness and architecture, highlighting the limitations of the Pd-NPs-enzyme interface. These findings provide mechanistic insight into ultrathin enzyme-electrocatalyst interfaces and are expected to impact the development of next-generation cholesterol biosensors.
Generating magnon frequency combs with tunable spacing via a single-frequency driving is crucial for practical applications, but it typically relies on complex spin textures like skyrmions or vortices. Here, we theoretically and numerically demonstrate magnon frequency comb generation in geometrically curved ferromagnetic thin films using single-frequency microwave excitation, without topological spin textures. We first show that the curvature transforms the planar ferromagnetic resonance into a localized, redshifted magnon bound state, which, under nonresonant driving, activates sequential three-magnon scattering processes assisted by the curvature-driven effective anisotropy and Dzyaloshinskii-Moriya interaction. It finally produces equally spaced, robust frequency combs with spacing exactly set by the bound mode frequency. Moreover, we find that the curvature gradient at the hybrid interface mimics an analog event horizon, with the bound state's redshift resembling gravitational effects in black hole physics. Micromagnetic simulations confirm these curvature-driven nonlinear phenomenon, unveiling a novel geometric strategy for controlling magnon interactions and advancing compact magnonic devices.
We propose a novel automatic approach to construct low-complexity, manifold, and non-self-intersecting cages, which preserves geometric features, maintains tight enclosure, enforces symmetry, and produces high-quality meshes. The method is realized through a coherent three-stage pipeline. Starting with isosurface extraction from the Signed Distance Field (SDF), our method inherently enforces envelope and non-self-intersection properties via density field modification and leverages sharp features of the input to drive a shape-preserving initial cage. Second, a constrained Quadric Error Metric (QEM) simplification problem is solved using a geometry-driven push-pull strategy, decimating the initial cage to a user-specified vertex count. This yields a concise intermediate structure while preserving the fundamental properties. Finally, we introduce a symmetry-aware refinement stage via an integrated optimization that simultaneously preserves symmetry, improves mesh quality, and ensures tight alignment with the input. We evaluate symmetry using a deviation metric (SDE) to extract a symmetry plane and produce a symmetry-consistent cage, followed by mesh quality enhancement and collision-free enforcement. Through extensive experiments on diverse models, including evaluations on public benchmarks, we demonstrate that our method outperforms existing techniques in handling complex shapes, automatically preserving features and symmetry while generating a compact, coarse cage.
Pollinator-mediated floral divergence is common, but the ecological mechanisms allowing divergent forms to coexist are often unclear. In South Africa, Lapeirousia anceps corolla tube length coevolves with the proboscis lengths of its local pollinators, long-proboscid flies. At most sites, proboscis and corolla length are closely matched, but there are occasional bimodal populations, where short- and long-tubed plants coexist. To understand the selective environment that may lead to bimodality and coexistence, we studied a bimodal population over a 21-year period, during which several fires occurred. Early in succession, pollinators and long-tubed plants were more abundant, and they produced more seeds than shorter-tubed plants. Later in succession, shorter-tubed plants produced relatively more seeds and flowered more frequently. In these plants, self-pollination contributed more to seed set, which explains their success in the absence of pollinators. Patterns of selection on tube length shifted in response to pollinator visitation rates, suggesting that the coexistence of short- and long-tubed plants might be facilitated by differences in post-fire successional niches. This illustrates how temporal heterogeneity caused by fire could promote co-occurrence of divergent forms. If so, ongoing changes in natural fire regimes may affect floral diversity.
Behavioural analysis with small aquatic model organisms is increasingly applied across diverse areas of biosciences and biomedicine, including ecology, pharmacology, and experimental neurobiology. High-throughput behavioural phenotyping workflows frequently employ organisms such as larval zebrafish (Danio rerio) in multiwell laboratory plates, yet the considerable cost of commercial imaging platforms often limits access for smaller laboratories, early career investigators and researchers in low- and middle-income countries. Here, we describe a rapid and practical approach for constructing an inexpensive, flexible and fully customisable behavioural data acquisition system suitable for use with a broad range of aquatic model species. We also examine an important but frequently overlooked methodological issue in quantitative behavioural research: how video acquisition parameters, particularly resolution and bitrate, influence the reliability of automated animal tracking. Using Artemia franciscana nauplii, adult Daphnia carinata, and 7 days postfertilisation larval Danio rerio, we show that video resolution is a major determinant of detection reliability, particularly for very small test organisms. Artemia franciscana could not be reliably tracked in multiarena configurations at resolutions below full high definition, whereas ultra high definition (3840 × 2160 pixels) provided robust detection across all tested species. We also show that concerns regarding ultra high definition file size can be mitigated by optimising bitrate settings. Moderate compression produced major reductions in file size while preserving detection reliability at practical compression levels. Together, these findings provide a practical framework for designing affordable and custom animal tracking systems and selecting video acquisition settings that support reliable high-throughput behavioural phenotyping across diverse aquatic species.
This study investigates the enhanced efficacy of photoinduced microbial inactivation targeting both planktonic and biofilm forms of Gram-negative bacteria, specifically Acinetobacter baumannii and Proteus mirabilis. The approach utilizes diaminocyclohexane (DACH)-based diamines to augment antibacterial photodynamic therapy (aPDT) employing 5-aminolevulinic acid. Results demonstrated that blue light irradiation produced the highest bactericidal effects, achieving mortality rates of 99.9990 ± 0.0008% for P. mirabilis and 99.990 ± 0.001% for A. baumannii at a light dose of 70 J·cm-2 in the presence of a non-cytotoxic concentration of (1R,2R)-N,N'-bis([1,1'-biphenyl]-4-ylmethyl)cyclohexane-1,2-diamine. The biofilm destruction efficiency was suboptimal, with maximum bactericidal efficiencies of 87.1 ± 0.1% and 73.1 ± 0.2% for P. mirabilis and A. baumannii, respectively, after exposure to blue light at a dose of 93.5 J·cm-2. Increased accumulation of protoporphyrin IX within bacterial cells was identified as a key factor contributing to the enhanced photobiocidal efficacy. Additionally, molecular docking analysis was employed to explore potential structural insights into the biological activity of this cyclohexane-1,2-diamine-based ligand. Computational simulations suggested that the ligand could plausibly accommodate within the protoporphyrin IX binding pocket of the modeled ferrochelatase, indicating a potential binding mode compatible with the catalytic cavity.
Choosing a relatively large reward with a lower chance of payoff compared with a relatively small reward with a higher chance of payoff can be operationalized as a risky choice. Risky choices are often observed in humans through gambling. Gambling disorder is characterized by excessive gambling that results in detrimental personal, social, and financial outcomes. Clarifying mechanisms that contribute to perseverative gambling could help clarify why problem gambling may occur. The presentation of win-paired cues is suggested to contribute to excessive gambling because of a conditioned-reinforcing function. Stimulant drugs have been shown to enhance conditioned reinforcers and to increase risky choice under certain experimental conditions. The present study was designed to examine how the combined administration of d-amphetamine, a commonly prescribed psychomotor stimulant, and the presentation of win-paired cues on winning and losing trials would impact risky choice patterns in a probability-discounting procedure in Sprague-Dawley rats. d-Amphetamine produced dose-dependent increases in risky choice with or without win-paired cues. Win-paired cues alone did not alter absolute levels of risky choice, but the combination of d-amphetamine and win-paired cues promoted perseveration for the risk-associated response option. This effect was particularly evident following losing trials where the win-paired cue was presented. Results are discussed in the context of electronic gambling machine use, and suggestions for research with more commonly used stimulants are offered.
Cell-penetrating peptides (CPPs) can deliver biomacromolecular cargos into cells, potentially enabling a new mode of intracellular drug delivery. However, a major problem with CPP-mediated delivery is entrapment of CPPs within endosomes as covalent linkages ensure CPPs and cargos share common fates. We previously developed a CPP-adaptor system based on reversible, calcium-dependent cargo binding that produces cargo release from adaptors as complexes dissociate following internalization and Ca2+ efflux from early endosomes. Having employed CPP-adaptors with an array of protein cargos of differing charges, it became apparent that positively charged cargos often appeared to dominate internalization and that association with the adaptor had little effect. To systematically address the effects of cargo charge and CPP function, we tested the ability of several adaptors to increase internalization of a set of adaptor binding GFP cargos having charges of +9, + 15, + 20, + 25 and +36. Intrinsic internalization of these cargos reproduced reported patterns showing that positive charge increases internalization. Interestingly, labeling these cargos with a chemical fluorophore revealed that GFP fluorescence grossly underestimated total internalization as shown by the fluor. Internalization was charge and concentration dependent with more positive cargos showing apparent saturation of internalization at 100-400 nM, well below the concentrations at which covalently linked CPP-cargos are commonly dosed. We tested the ability of 5 adaptors to internalize these cargos. Our prototype adaptor, TAT-CaM, was completely ineffective with the + 9 cargo, but internalized moderately charged cargos extremely efficiently at concentrations far below the µM range. A derivative adaptor, TAT-LAH4-CaM, was highly effective with all cargos and produced similar maximal internalization at 100-400 nM. However, two adaptors specifically designed with increased positive charge inhibited internalization of the most positive cargos. One of these, GFP-CaM, based on the supercharged GFP with net charge of +36, did increase internalization of the least positive cargos, demonstrating an adaptor with high affinity for the cell surface can increase internalization of a neutral cargo at very low concentration. The common maximal level of intrinsic GFP cargo internalization correlated with surface loading of these cargos, suggesting a limit to the beneficial effects of increased plasma membrane association. However, TAT-CaM further increased internalization above intrinsic levels via an apparently distinct mechanism. In this limited study of the interaction of cargo charge and adaptor efficacy, we found diverse behaviors that hint at the power and flexibility possible with adaptor/cargo internalization.
Diabetes mellitus is a chronic metabolic disorder characterized by persistent hyperglycemia and oxidative stress, leading to multi-organ dysfunction and impaired wound healing. This study evaluated the antidiabetic, antioxidant, hematological, biochemical, and wound-healing effects of silver-doped copper carbonate nanoparticles (Ag-CuCO₃ NPs) in alloxan-induced diabetic albino mice. Diabetes was induced via intraperitoneal injection of alloxan monohydrate (150 mg/kg) and animals were divided into control, diabetic control, and treatment groups receiving low dose (LD) or high dose (HD) of Ag-CuCO₃ NPs orally for 28 days. Hematological, biochemical, hormonal, electrolyte, and wound-healing parameters were assessed, and data were analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post hoc test, with p < 0.05 considered statistically significant. Diabetic controls showed severe hyperglycemia (> 280 mg/dL), elevated LDL (64 ± 3 mg/dL), reduced RBC count (5.1 ± 0.3 × 10⁶/µL) and delayed wound closure beyond Day 20. Low-dose Ag-CuCO₃ NP treatment significantly reduced fasting blood glucose levels from diabetic values (> 280 mg/dL) toward near-normal levels and improved hematological indices, including a ~ 42% increase in RBC count (2.85 to 4.05 × 10⁶/µL), ~ 38% increase in hemoglobin (7.46 to 10.33 g/dL), and ~ 115% increase in hematocrit (10.43 to 22.43%) compared with diabetic controls. Insulin levels increased by ~ 53% (6.12 to 9.35 µIU/mL), while sodium levels improved by ~ 3% (136.13 to 140.03 mmol/L), indicating partial restoration of metabolic and renal balance. High-dose treatment produced a more pronounced improvement in oxidative status, with SOD activity increasing by ~ 100% (2.31 to 4.62 U/mg protein) and lipid peroxidation marker MDA decreasing by ~ 51% (6.41 to 3.12 nmol/mg protein). However, high-dose treatment was also associated with elevations in hepatic enzymes, including AST (~ 50% increase; 56.33 to 84.66 U/L) and ALT (~ 114% increase; 25.66 to 55 U/L), suggesting potential dose-related hepatic stress. While high-dose treatment achieved faster glycemic and oxidative stress correction, low-dose treatment provided more balanced therapeutic efficacy with minimal biochemical disturbances. Overall, these findings provide preliminary evidence of dose-dependent antidiabetic, antioxidant and wound-healing effects of Ag-CuCO₃ nanoparticles in a preclinical diabetic model, warranting further mechanistic and comparative investigations.
Klebsiella pneumoniae (K. pneumoniae) is a bacterium that causes the production of thick, viscous mucus. Lower respiratory tract infections caused by K. pneumoniae are characterized by the presence of thick mucus. It can also lead to surgical site infections, urinary tract infections (UTIs), diarrhea, upper respiratory tract infections, wound infections, meningitis, bacteremia, and septicemia. This study aimed to determine the phenotypic and genotypic ESBL production, antibiotic susceptibility, and plasmid-mediated quinolone resistance (PMQR) genes in 192 K. pneumoniae strains (55.2% urine) isolated from clinical samples in hospitals in Konya province. Phenotypic ESBL production was assessed using the double disc synergy test. The presence of blaCTX-M, blaTEM, blaOXA, and blaSHV genes was investigated by multiplex PCR. PMQR genes (qnrA, qnrD, qnrB, qnrS, oqxAB, aac(6')-Ib-cr, qepA, qnrC) were also screened by multiplex PCR. Among the 192 isolates, 36 (18.75%) were phenotypically ESBL-positive and 186 (96.87%) were genotypically positive. Of the 19 antibiotics tested, the highest resistance was to Ampicillin (10 µg) (97.9%), and the lowest to Colistin (10 µg) (11.45%). All strains were resistant to at least one antibiotic, with 113 different resistance profiles identified. According to the multiple antibiotic resistance (MAR) index, 72% (139) of strains had an index > 0.2. The most prevalent ESBL gene was blaSHV (184; 95.83%), followed by blaTEM (95; 49.47%), blaOXA (87; 45.31%), and blaCTX-M (27; 14.06%). The most common ESBL gene combination was blaSHV + blaTEM + blaOXA (26.5%). Among PMQR genes, oqxAB was most frequent (156; 81.25%), while qnrC was least frequent (7; 3.6%). These findings highlight the importance of detecting ESBL-producing strains and PMQR genes for effective therapy. Rapid identification using multiplex PCR may help prevent prolonged hospitalization and improve patient outcomes.
Drug-induced pulmonary toxicity (DIPT) is a serious and often underappreciated complication that can emerge in patients receiving a wide variety of prescribed drugs, from cytotoxic chemotherapy agents and antiarrhythmics to immunomodulatory compounds. Four agents, in particular methotrexate (MTX), cyclophosphamide (CPA), bleomycin (BLM), and amiodarone (AMI) have the most extensive body of evidence documenting their pulmonary adverse effects, with each capable of producing distinct injury patterns that nonetheless share overlapping features. The mechanisms driving DIPT are far from simple; multiple pathways converge to produce the damage, including oxidative stress, dysregulated inflammatory responses, various modes of programmed cell death (apoptosis, pyroptosis, and ferroptosis), and activation of pro-fibrotic cascades most notably TGF-β1/Smad2/3, NF-κB, and the NLRP3 inflammasome. Standard treatment options for early drug withdrawal, corticosteroids, and supportive care are frequently inadequate, and a significant proportion of patients are left with persistent pulmonary fibrosis long after the causative drug has been stopped. This review offers a thorough examination of the epidemiology, risk determinants, pathological features, diagnostic criteria, and molecular underpinnings of DIPT as caused by MTX, CPA, BLM, and AMI. Going further, it assembles a structured, evidence-grounded compilation of the pharmacological and natural agents that have shown protective potential in experimental DIPT models. Four summary tables covering a combined total of 74 protective interventions are presented, detailing experimental designs, induction protocols, and mechanistic targets. The striking range of strategies assessed spanning small-molecule antioxidants and plant-derived polyphenols to repurposed established drugs highlights the growing understanding that DIPT's mechanistic complexity calls for multi-targeted therapeutic solutions. Taken together, this review integrates available preclinical and clinical data into a practical framework for developing rational lung protection strategies against drug-induced injury.
Conventional laparoscopic instruments provide limited dexterity in confined anatomical environments, whereas existing articulated designs rely on complex rigid joints or motorised actuation, increasing mechanical complexity and limiting miniaturisation. A hybrid compliant-tendon laparoscopic instrument was developed to integrate articulated distal motion and active grasping within a 5 mm form factor. The compliant segment was fabricated from super-elastic Nitinol, and grasping was achieved using a tendon-pulley mechanism. The system was evaluated using nonlinear finite element analysis and experimental testing, including digital image correlation and benchtop force measurements. The mechanism achieved up to 75° articulation with mean model errors below 8%. The grasper produced a maximum force of 13.2 ± 0.47 N and maintained performance across articulation angles. Pull-out tests confirmed stable interaction with soft-tissue analogues. The proposed design demonstrates a compact, low-complexity approach for combining compliant articulation and tendon-driven grasping within a 5 mm surgical instrument.
Haloferax volcanii DSM27206 (formerly Haloferax alexandrinus TMT) is a halophilic red archaeon. The genome contains a circular chromosome (2,850,539 bp with a guanine-cytosine [GC] content of 66.7%) and three plasmids (5,595, 296,275, and 496,018 bp with a GC content of 64.0, 66.4, and 64.0%, respectively). National Center for Biotechnology Information Prokaryotic Genome Annotation Pipeline annotation identified 3,458 protein-encoding genes.
Some drugs possess reinforcing properties, which are commonly studied using the conditioned place preference (CPP) paradigm. CPP has been applied across a wide range of taxa, including nonmammalian vertebrates and invertebrates. In the present study, goldfish were used as an experimental model to examine CPP induced by nicotine, a substance that remains a major public health concern. Nicotine produced CPP at lower doses and conditioned place aversion (CPA) at higher doses. One advantage of fish models is the feasibility of drug administration by immersion, an animal-friendly alternative to injection. However, because intraperitoneal injection is the standard method in rodent studies, quantitative comparisons of dose-response relationships across taxa are difficult. To address this issue, I compared the dose-response relationship of general activity following nicotine administration by immersion with that following intraperitoneal injection in goldfish. Based on this comparison, a conversion factor between the two administration procedures was estimated. Using this conversion, nicotine-induced CPP and CPA in goldfish were quantitatively compared with findings from rodent studies. These results contribute to cross-species evaluation of drug reinforcement and support the utility of fish models in behavioral pharmacology.
Agricultural systems are designed to meet the increasing global demand for food. However, pests pose a significant threat to agricultural productivity, potentially hindering the ability to meet the demand. Biological control is considered an effective method to manage these pests and tackle food loss. Among the various biological agents, endophytic fungi stand out as a promising alternative to insecticides due to their numerous beneficial effects. In this review, we summarize the reports on the major phyla and classes of endophytic fungi as well as their identified sources. Endophytic fungi are known to synthesize insecticidal metabolites that directly help to prevent pest invasion, reduce the pest's adaptability to their surroundings, and increase their sensitivity to natural enemies. Besides, several metabolites produced by the endophytic fungi signal the host plant to produce its defense chemicals, triggering the plant's immune response and thereby enhancing its resistance. Therefore, the review further discusses the direct (endophyte-insect interactions) and indirect effects of the endophytic fungi on the insect pests (enhanced plant defense) and provides evidence to support their significant role in the endophytic fungus-plant-insect triadic relationship. Taking these reports into consideration, we suggest that endophytic fungi could serve as a significant component of sustainable pest management. However, the negative impact of the fungal metabolites on the mammals that consume the host plants needs to be investigated.
Macrophages are central to host immunity and tissue homeostasis, exhibiting remarkable functional plasticity across a continuum of states-ranging from pro-inflammatory (M1-like) to anti-inflammatory and tissue-reparative (M2-like) phenotypes. Environmental exposures can induce persistent epigenetic changes that shape macrophage responses well beyond the acute phase, a phenomenon now recognized as trained immunity. This narrative review synthesizes current knowledge on how diverse components of the exposome-including diet, air pollution, agricultural chemicals, heavy metals, endocrine-disrupting chemicals, per- and polyfluoroalkyl substances (PFAS), alcohol, smoking, the gut microbiome, maternal diet, and psychosocial stress-remodel the macrophage epigenome. We examine the underlying epigenetic mechanisms, namely DNA methylation, histone modifications, and non-coding RNAs, and discuss their impact on macrophage polarization, cytokine production, and trained immunity induction. Special emphasis is placed on the distinction between bona fide trained immunity and transient inflammatory skewing, the limitations of the classical M1/M2 framework, and the identification of "epigenetic vulnerability nodes" at which multiple environmental signals converge on a small set of chromatin-modifying enzymes and transcription factors. We also highlight critical knowledge gaps, including the lack of data for emerging contaminants such as micro- and nanoplastics, the uncertain reversibility of exposure-induced epigenetic marks, and the challenge of demonstrating transgenerational inheritance in humans. By connecting molecular mechanisms with broader public health implications, this review provides a critical framework for understanding environmentally driven immune dysregulation and outlines future research directions, including mixture toxicology, single-cell multi-omics, and the integration of epigenetic endpoints into chemical risk assessment.
The purpose of this study was to compare the effectiveness of Low-Level Laser Acupuncture with Dry Needling along with Routine Physical Therapy on clinical and functional outcomes in patients with Chronic Cervical Myofascial Pain Syndrome. An Assessor-blind Randomized Controlled trial was conducted at the Physical Therapy department of Rawal General & Dental Hospital, Pakistan. The study duration was June 2023 to November 2024. A non-probability convenience sampling technique was employed to produce a study sample of 100 individuals with Chronic Cervical Myofascial Pain Syndrome. The 100 eligible participants, with a mean age of 34 ± 7.21 years, were randomly allocated into two equal groups (n = 50 per group). Group A received Low-Level Laser Acupuncture and Routine Physical Therapy, while Group B underwent Dry Needling and Routine Physical Therapy. The outcomes of the study were pain intensity, neck disability, cervical range of motion, and health-related quality of life measured by the Numeric Pain Rating Scale, Neck Disability Index, Goniometer, and SF-36 health survey questionnaire, respectively. The outcome measures were evaluated at three temporal points: the pre, mid, and post-treatment sessions. Each participant received a total of 18 treatment sessions. The IBM SPSS Statistics version 26.0 was used to analyze and interpret the results. The Mann-Whitney U test, Friedman test, and subsequent Wilcoxon signed-rank post hoc analyses with Bonferroni adjustment demonstrated statistically significant differences (p < 0.05 in all cases), both intergroup and intragroup over time. The Low-Level Laser Acupuncture and Dry Needling both proved to be efficient adjunct therapies to routine physical therapy for Chronic Cervical Myofascial Pain Syndrome for different durations of improvement.
There is a growing need in biochemistry education to move beyond traditional, recipe-based experiments and toward integrated, research-based experiences that mimic real-world scientific inquiry. We designed a multi-week, project-based learning (PBL) module where students undertake the complete process of producing ergothioneine (ERG), a valuable antioxidant, using recombinant DNA technology in Escherichia coli. The project encompasses molecular cloning (optional), protein expression analysis, fermentation in a bioreactor, and product quantification. The module was successfully implemented in an upper-level biochemistry laboratory course. Assessment via pre- and post-lab quizzes, lab reports, and a comprehensive student self-evaluation survey showed significant gains in technical skills, conceptual understanding, and research confidence. This laboratory module provides an effective framework for integrating modern biotechnological techniques into the undergraduate curriculum. It enhances student learning by contextualizing theoretical knowledge within a continuous, authentic research project, thereby fostering critical thinking, problem-solving skills, and enthusiasm for biochemical research.