Search Results (3,868)

Human papillomavirus type 53 (HPV53) is one of the most prevalent HPV genotypes in China, frequently detected in cervical intraepithelial neoplasia and cervical cancer, yet remains outside the coverage of all currently available prophylactic vaccines and is relatively understudied. This study performed a comprehensive analysis of HPV53 clinical infection profiles, genomic diversity, and T-cell epitopes to inform therapeutic vaccine development. Clinical analysis of 158 HPV53-positive patients showed that infections were most prevalent in women aged 40–59 years, with persistent infection identified in 13.3% participants and a subset of cases associated with cervical lesions. Genomic analysis of 134 HPV53 isolates identified four lineages (A-D, with lineage D further subdivided into four sublineages, and an overall nucleotide variability of 4.4%. E2 was the most variable protein while E7 was the most conserved. Immunoinformatic prediction identified 176 HLA class I-restricted T-cell epitopes across E6, E7, E1, and E2, from which 20 candidates were selected for experimental validation. Ten demonstrated strong HLA binding affinity in vitro, and murine immunization identified a E6 peptide VYNFAYTDL as an immunodominant epitope. Three validated epitopes exhibited sequence overlap with 12 to 13 of other 13 high-risk HPV genotypes, suggesting their potential as broadly cross-reactive targets. These findings clarify the genomic diversity and immunogenic epitope landscape of HPV53, providing a foundation for the rational design of therapeutic vaccines. Full article

Objectives: This study aimed to develop curcumin nanoparticles (Cur@PCL-PEG-MF/cRGDfc) with retinal-targeting capability and to evaluate their biological effects and pharmacological mechanisms in vitro. Methods: After synthesis of the carrier framework, metformin (MF) and cRGDfc were conjugated to the carrier material using the carbodiimide method and Michael addition reaction, respectively. Subsequently, self-assembled nanoparticles were formed from the carrier and curcumin under specific conditions. The materials were characterized by spectroscopy, chromatography, elemental analysis, energy-dispersive spectroscopy and X-ray diffraction. The efficacy of the formulation was evaluated in two cell lines, ARPE-19 and HUVEC-T1. In addition, the pharmacological mechanism was explored using transcriptome sequencing as a complementary approach. Key Findings: Self-assembled nanoparticles were successfully prepared by combining the two modified carrier materials, PCL-PEG-MF and PCL-PEG-cRGDfc, with curcumin. The nanoparticles exhibited an encapsulation efficiency of 78.09%, a particle size of 162.33 nm, and a zeta potential of −23.28 mV and displayed a spherical morphology. They showed sustained release in simulated physiological conditions and stronger affinity for ARPE-19 cells under oxidative stress. Nearly 100% of the nanoparticles were internalized by the cells, which was accompanied by reduced ROS and LDH release and decreased DNA fragmentation. In addition, the nanoparticles inhibited neovascularization by reducing VEGF-A release, thereby potentially protecting the retina in macular degeneration and reducing choroidal hemorrhage. Further analyses showed that curcumin and its nanoformulations significantly reduced the expression of inflammatory factors such as IL-1β and IL-18, lowered the protein levels of Caspase-1, GSDMD-N, and NLRP3, and increased AMPK levels. Conclusions: Using PCL-PEG as the carrier framework, MF and cRGDfc were conjugated to construct a curcumin-loaded nanoparticle with retinal-targeting capability. This nanoparticle, characterized by a small particle size, sustained release, and targeted delivery to retinal pigment epithelium (RPE) cells under oxidative stress, alleviated oxidative stress-induced damage. Its therapeutic effect may be mediated, at least in part, by interference with the AMPK/mTOR pathway and activation of the NLRP3/Caspase-1/GSDMD pathway. Full article

Background: Olfactory dysfunction is a pathology associated with viral infections, toxic damage, aging, and neurodegenerative diseases. Damage to the olfactory epithelium impairs olfactory function and related neurological behaviors. This study evaluated the restorative effects of Scutellaria baicalensis Georgi (SBG) extract and baicalein in a methimazole-induced olfactory dysfunction model. Methods: Olfactory epithelial damage was induced in mice with methimazole, followed by treatment with SBG extract or baicalein. Olfactory and neurobehavioral functions were assessed using odor-finding, novel object recognition (NOR), Morris water maze (MWM), open field (OFT), and elevated plus maze tests (EPM). Histological, immunohistochemical, and in vitro analyses were performed to evaluate epithelial regeneration, mature olfactory sensory neurons (OSNs) expressing olfactory marker protein (OMP), and proliferative activity. Results: Methimazole induced severe olfactory epithelial damage, impairing olfactory behavior and reducing learning and memory. Treatment with SBG extract and baicalein significantly improved olfactory and cognitive functions. Histological and immunohistochemical analyses confirmed restoration of epithelial structure and olfactory neurons. In vitro, SBG extract increased epithelial cell density and modulated proliferative activity. Conclusions: SBG extract and baicalein promote recovery of olfactory function and improve neurobehavioral outcomes, indicating their potential as therapies for olfactory dysfunction. Full article

This study investigates color and surface alterations in neat and modified PEEK materials (10 × 3 mm, n = 140) immersed in various beverage solutions. Surface roughness (Ra) and color change (ΔE) were analyzed, supported by SEM and AFM evaluations. Specimens polished with 400–1200 grit sandpaper were measured via profilometry before a 30-day immersion in distilled water, coffee, tea, cola, and red wine (refreshed every 12 h). Post-immersion results indicated that material and solution types significantly influenced Ra and ΔE values (p < 0.05), with the TP group being the least affected. Statistically, tea caused the greatest discoloration. The GFP group exhibited the highest Ra, indicating inferior surface stability, whereas TP and CR groups remained below the 0.2 μm clinical threshold. Specifically, the TP group demonstrated the lowest Ra value. Although perceptible color changes occurred in all materials, a positive correlation was identified between material type and beverage solutions. SEM and AFM analyses confirmed the presence of surface micro-cracks and morphological irregularities. Full article

The valorization of culinary by-products into functional bioactive resources represents a significant advancement in sustainable biotechnology. This study characterizes an extract derived from “battuto toscano” by-products, a traditional blend of garlic, onion, carrot, and celery trimmings, recovered through circular economy principles. Comprehensive antioxidant profiling was performed alongside biological evaluations on human cell lines and anti-glycation assays. Results from Folin–Ciocalteu, FRAP, and TEAC assays confirmed a high concentration of secondary metabolites with significant scavenging capacity. In vitro testing on primary human fibroblasts and HaCaT keratinocytes revealed a concentration- and time-dependent biological response, with lower concentrations showing better compatibility and transiently enhancing HaCaT metabolic activity. Furthermore, BTE reduced AGE-associated fluorescence in the BSA–glucose model, particularly at 5 mg/mL, supporting its potential anti-glycation activity. These findings establish “battuto toscano” by-products as a reservoir of sustainable biomolecules. This study offers a transformative resource for the pharma/nutraceutical sectors by bridging culinary tradition with biomedical innovation. Full article

Oxidative stress is a major contributor to the development of age-related macular degeneration (AMD), and excessive oxidative stress can induce retinal pigment epithelium (RPE) dysfunction, apoptosis, and retinal degeneration. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) is a major enzymatic source of reactive oxygen species (ROS); however, its mechanistic role in sodium iodate (NaIO₃)-induced oxidative injury remains unclear. Tetrahydrocurcumin (THC), the major metabolite of curcumin, exhibits potent antioxidant and cytoprotective activities, but its protective effects against AMD-associated retinal degeneration have not been fully elucidated. In the present study, we investigated whether THC protects against NaIO₃-induced ROS-mediated apoptosis in RPE cells through regulation of NOX2 signaling. In vitro, THC significantly attenuated NaIO₃-induced cytotoxicity and prevented apoptosis by suppressing hydrogen peroxide (H₂O₂) production and intracellular ROS accumulation in ARPE-19 cells. THC also preserved mitochondrial membrane potential by inhibiting the Src/p47^phox/NOX2 signaling pathway and subsequently attenuated mitochondria-mediated apoptotic signaling. Furthermore, THC markedly reduced the expression of apoptotic proteins, including Bax, cleaved caspase-3, and cleaved PARP, concomitantly with suppression of Ras/Raf/MEK/ERK signaling. Mechanistically, treatment with the selective NOX2 inhibitor GSK2795039 significantly attenuated NaIO₃-induced ROS accumulation and mitochondrial depolarization, while co-treatment with THC further enhanced these protective effects. In vivo, THC ameliorated NaIO₃-induced retinal structural abnormalities by preserving the outer nuclear layer (ONL), reducing caspase-3 expression, and improving pupillary light responses in mice. Collectively, these findings demonstrate that THC protects against NaIO₃-induced retinal degeneration through suppressing NOX2-dependent oxidative stress and downstream Ras/Raf/MEK/ERK-mediated apoptotic signaling, highlighting its potential as a therapeutic candidate for AMD and other oxidative stress-related retinal disorders. Full article

Recent evidence suggests that microbiota-derived porphyrins contribute to skin aging, a phenomenon termed porphyr’aging. These pro-inflammatory molecules alter the expression of genes involved in senescence, trigger melanogenesis, and decrease collagen I synthesis in skin. The aim of this study was to evaluate the anti-aging properties of an upcycled Laminaria hyperborea extract (LHE) targeting bacterial porphyrins discovered after screening. The impact of LHE on porphyrin biosynthesis and on melanogenesis and wrinkles was evaluated using in vitro and ex vivo tests and by conducting a double-blinded vs. placebo clinical trial. LHE significantly reduced coproporphyrin III production in Gram-positive skin bacteria and significantly decreased porphyrin levels in vivo at the skin surface. This activity was supported by a specific composition of LHE, comprising laminaran and mannitol. It also significantly decreased melanin content in skin explants and pigmentation in the clinical study (−5.9%). This effect was particularly pronounced in dark spots (ITA +39.9%), and the number of precursor spots also decreased (−6.9%). In addition, LHE significantly stimulated type I α-1 pro-collagen production in fibroblasts and increased collagen I and elastin expression in skin explants. These results were consistent with the clinical study, showing significant reductions in wrinkle number (−9.8%) and area (−5.8%). These findings suggest that targeting microbiota-derived porphyrins and their consequences may represent a promising approach to reduce the visible signs of aging. Full article

Background: Bone and joint infections (BJIs) are a significant clinical challenge due to their tendency to recur, increased healthcare expenses, reduced quality of life, and mortality. Patients with BJIs are a heterogeneous group due to their different clinical presentations as well as patient-related risk factors. Empiric antibiotic regimens are commonly based on deductions from in vitro microbiologic findings, despite the fact that their relative efficacy and optimal antibiotic choices are underexplored. Methods: This retrospective cohort study included 521 patients surgically treated for BJIs at a specialized orthopedic infection unit between 2016 and 2023. Treatment strategies were guided by the Oral Versus Intravenous Antibiotics for Bone and Joint Infection (OVIVA) trial. All patients received a narrow-spectrum Gram-positive–targeted empiric systemic antibiotic regimen determined according to regional recommendations in collaboration with infectious disease specialists. The primary outcome was clinical failure within one year, with a minimum follow-up of 12 months. For the analyses, the patients were divided into three groups based on microbiological susceptibility: susceptible (SusEmp), non-susceptible (NonSus) and culture-negative (CulNeg) patients. Results: The three groups were found to differ significantly in seven patient-related factors: sex, age at primary operation (OP age), BMI, ASA group, diabetes status, peripheral arterial disease status (PAD), and endocrinopathy status (other than diabetes). In performing multivariate analyses, OP age was found to be independently associated with the overall failure rate (p = 0.04) and ASA group (p = 0.047), and PAD (p = 0.043) was independently associated with the secondary outcome of proximal amputation. Patients with non-susceptible pathogens (NonSus) had more than twice the odds of clinical failure (OR: 2.10; 95% CI: 1.12–3.95) and nearly fivefold higher odds of secondary proximal amputation (OR: 4.95; 95% CI: 1.41–23.2) compared with patients with susceptible pathogens (SusEmp). Conclusions: The current study demonstrates that a large group of patients can presumably be treated safely with a more restrictive narrow approach. More studies are needed to identify subgroups suited for the safe use of a narrow-spectrum empiric regimen, hereby reserving the broad-spectrum antibiotics for patients with the right indications and for whom it would have a positive effect on the clinical outcome. Such an approach would justify a more restrictive stewardship of broad-spectrum antibiotic use without negatively impacting patient outcomes. Full article

Background/Objectives: Embryo implantation is a highly regulated, multistep process requiring precise synchronization between a developmentally competent blastocyst and a receptive endometrium. Despite advances in reproductive medicine, implantation failure remains a major limiting factor in assisted reproductive technology (ART), particularly in cases of recurrent implantation failure (RIF). This review aims to summarize current knowledge on the molecular, cellular, and immunological mechanisms governing embryo–endometrial interaction and to evaluate contemporary strategies for optimizing implantation outcomes. Methods: This narrative review synthesizes the current literature on embryo implantation, including studies addressing uterine receptivity, etiological factors contributing to implantation failure, and emerging diagnostic and therapeutic approaches. The review integrates findings from molecular biology, clinical ART practices, and bioengineering-based models. Key areas include transcriptomic tools such as endometrial receptivity analysis, time-lapse imaging, artificial-intelligence-based embryo selection, and advanced in vitro models (e.g., microfluidic “womb-on-a-chip” systems and three-dimensional embryo–endometrial platforms). The literature was identified through major biomedical databases, following a structured but non-systematic approach. Results: Implantation success is dependent on a complex interplay of hormonal regulation, gene expression, immune modulation, and embryo quality. Disruption of uterine receptivity during the window of implantation is a critical contributor to infertility and RIF. Multiple factors—including genetic abnormalities, maternal age, lifestyle influences, immunological imbalance, uterine pathology, and chronic endometrial conditions—are implicated in implantation failure. Emerging technologies, such as AI-assisted embryo selection, transcriptomic profiling, and advanced in vitro implantation models, provide enhanced insight into implantation dynamics and offer potential for improved clinical outcomes. Conclusions: Advances in understanding embryo implantation and the development of innovative diagnostic and therapeutic technologies hold significant promise for improving reproductive success. However, further research, validation, and standardization are required before these approaches can be fully integrated into routine clinical practice. A more personalized and mechanism-based approach to implantation may ultimately enhance ART outcomes and reduce the burden of infertility. Full article

Invasive candidiasis caused by drug-resistant Candida species represents a critical global health challenge, with few novel therapeutic scaffolds under development. Here, silver nanoparticles were synthesized using a 21-day fermented Chun Mee kombucha tea extract (K-AgNPs) and characterized by UV-Vis spectroscopy, transmission electron microscopy, nanoparticle tracking analysis, and Fourier-transform infrared spectroscopy. LC-MS/MS profiling of the kombucha substrate documented a phytochemical landscape dominated by epigallocatechin (up to 122,631 µg/mL) and epigallocatechin gallate (up to 415 µg/mL), with a progressive ~80% decline in epicatechin and concomitant increases in gallic acid and chlorogenic acid across the 21-day fermentation. K-AgNPs obtained were spherical, 19.4 nm (±7.9 nm SD) in diameter, with a surface plasmon resonance peak at 415 nm. FTIR confirmed phenolic, carboxylate, and glycosidic surface capping. Antifungal susceptibility testing against eight Candida species, including the WHO critical–priority pathogen Candidozyma auris, showed concordant minimum inhibitory and minimum fungicidal concentrations of 0.80–1.60 µg/mL, confirming fungicidal activity. In vivo evaluation in Galleria mellonella larvae across six infection models demonstrated that K-AgNP treatment at the species-specific MIC significantly improved larval survival versus untreated infected controls (p < 0.01–0.001), while nanoparticle-only groups maintained ≥98% survival, indicating negligible toxicity. Co-treatment amplified total hemocyte mobilization, and K-AgNP-only larvae maintained hemocyte viability above 96% at all time points, indistinguishable from negative controls. Together, these findings demonstrate antifungal activity of K-AgNPs across the genus Candida in standardized in vitro and in vivo settings and provide justification for further investigation, including head-to-head comparison against licensed antifungals and physicochemical validation of nanoparticle stability under assay conditions. Full article

Accumulation of aggregated amyloid beta (Aβ) species is a defining pathological hallmark of Alzheimer’s disease and is associated with extensive neuronal structural abnormalities. Mild cognitive impairment (MCI), a transitional stage between normal aging and the onset of dementia, is thought to represent an early phase of this pathological continuum. Studies at the cellular level suggest that the conditions impair the maintenance of established neuronal processes/networks and restrict their capacity for elongation or re-elongation. They may also attenuate the activation and process extension of quiescent neural progenitor or stem-like cells. These early cellular changes precede overt neurodegeneration in neural tissue and are likely to contribute to cognitive decline. They highlight the importance of in vitro models for identifying molecular targets involved in recovery from disease. In this study, we investigated the effects of aggregated Aβ (25–35) on neuronal process elongation and associated intracellular events in the N1E-115 cell line, a widely used model of neuronal differentiation. Addition of aggregated Aβ to cultured N1E-115 cells attenuated process elongation in a concentration-dependent manner. This morphological impairment was accompanied by decreased expression of neuronal differentiation markers. In contrast, at the half-maximal inhibitory concentration for process elongation, long-term cultured cells did not exhibit apparent process retraction or degenerative morphology. This mild but progressive impairment, without extensive cell death, is consistent with the cellular features of early-stage conditions rather than advanced Alzheimer’s pathologies. Similar results were observed in primary cortical neurons. Aβ also decreased the level of GTP-bound Ras and phosphorylation of the downstream mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK). Furthermore, treatment with hesperetin, a bioactive flavonoid compound, recovered the Aβ-induced inhibition of neuronal process elongation. Hesperetin also restored Ras and MAPK/ERK states, suggesting that its effects are associated, at least in part, with modulation of signaling through Ras and MAPK/ERK. Our findings suggest that hesperetin may serve as a useful molecular probe for modulating early cellular responses associated with Alzheimer’s disease-related pathology. This in vitro model might serve as a useful platform for investigating the molecular target candidates involved in recovery from nervous system disorders. Full article

Effective hand washing takes time and hand sanitizers that contain alcohol have a number of drawbacks, and frequent use of alcohol may cause skin damage. The objective of this study is to formulate nanostructured lipid carrier systems containing chlorhexidine digluconate to be applied topically for hand hygiene, especially for people sensitive to alcohol. A cytotoxicity experiment was conducted to ascertain the safe dosage for each of the three nano-cream formulas (F1, F2 and F3). Following each treatment, the viral titer was assessed using tissue culture infectious dose₅₀ and standard plaque assays. The selected formulation was characterized rheologically. Furthermore, fifteen volunteers of various ages and genders participated in the vivo antimicrobial test of the selected formulation as a hand sanitizer. All of the formulas were found to be safe. Using the disc diffusion method, the three formulations exhibited in vitro antimicrobial effects against different microbes. F1 showed biphasic release, reasonable skin deposition and spherical droplets under a microscope. F1 exhibited a non-Newtonian shear thinning flow behavior. After 30 min, the reduction values for rotavirus and Phix-174 were 21 and 4%, respectively. Additionally, the impact of F1 was assessed on the infectivity of simian rotavirus sa-11 (ds RNA) and Phix-174 (ss DNA) bacteriophage. According to the findings of the in vivo study, the percentage of total bacterial counts that were removed varied from 91 to 100%. Moreover, the range of the removal percentage of total fungi was 95.38 to 100%. In summary, F1 can be used as an economic, safe, and effective hand antiseptic. It can also completely replace alcohol in the market. Full article

Fucoidan, a sulfated polysaccharide, is known for its beneficial bioactive effects, for example antioxidant, anti-inflammatory, and vascular modulatory effects. Such a bioactive compound may also be useful for treating neurodegenerative diseases like age-related macular degeneration (AMD). Our research focuses on AMD-related pathomechanisms using primary porcine retinal pigment epithelium (RPE) cells in vitro and zebrafish (Danio rerio) models in vivo. We tested the bioactivity of a commercially available fucoidan (FVs) from bladderwrack with regard to pathomechanisms of AMD. We performed multiplex assays, RT-qPCR and fluorescence-based assays for the formation of nitric oxide (DAF-FM assay) and reactive oxygen species (DCF-DA assay) to analyze angiogenesis-related chemokines and pro-inflammatory cytokines as well as protection against oxidative stress and inflammatory insult. Our results showed that FVs significantly reduced the secretion of pro-angiogenic vascular endothelial growth factor A (VEGF-A) and follistatin as well as the pro-inflammatory cytokines interleukin 8 (IL-8) after lipopolysaccharide (LPS) and polyinosinic/polycytidylic acid (PIC) induction. Interleukin 6 (IL-6) was also reduced in the supernatant of the RPE cells. Additionally, in zebrafish, fucoidan decreased the production of NO and ROS. Gene expression of zebrafish embryos revealed anti-inflammatory effects by suppressing pro-inflammatory genes and significantly downregulating, e.g., interleukin 1 beta (IL-1β). These findings indicate modulation of oxidative stress, inflammatory responses, and VEGF secretion of the used FVs. This study demonstrates that fucoidan possesses AMD-relevant bioactivities in vitro and in vivo, suggesting fucoidan warrants further investigation in AMD-related research and related pathological mechanisms. Full article

Prolonged use of clear aligners promotes bacterial colonization and biofilm formation, which can compromise orthodontic outcomes. There is a clear clinical demand for approaches that can suppress pathogenic activity while preserving the fundamental functional and material characteristics of the aligners. To address this need, a novel strategy of fabricating metal–organic framework (MOF) coatings on aligners was adopted. Metal–organic frameworks (MOFs) have emerged as promising antibacterial coating materials by combining antimicrobial metal ions with biocompatible organic ligands. Three distinct porphyrin-based MOFs (Ag-, Cu-, and Ce-TCPP) were synthesized and fabricated as coatings on clear aligner surfaces via a coordination-driven self-assembly approach. The coated aligners were comprehensively assessed in vitro to determine their antibacterial performance, anti-inflammatory potential, biocompatibility, and key physical characteristics. Among the three coatings, Ag-TCPP showed the most favorable overall antibacterial and anti-biofilm performance in the present experimental system and facilitated macrophage polarization toward an anti-inflammatory M2-like phenotype. Ag-TCPP exhibited a significant inhibition zone of 6.75 ± 0.25 mm and reduced biofilm biomass by 72.2%. All MOF coatings exhibited excellent biocompatibility, and their application did not compromise the aligners’ mechanical integrity or aesthetic properties (light transmittance). This study reports the successful development of a novel metal–organic framework (MOF)-based coating strategy for clear aligners. Among the formulations investigated, the Ag-TCPP coating exhibited outstanding antibacterial and immunomodulatory performance while maintaining the critical mechanical integrity and aesthetic qualities of the aligner. The findings of this work offer a practical approach to designing multifunctional orthodontic devices that may reduce biofilm-related complications and improve clinical outcomes. Full article

Background: The purpose of this in vitro study was to evaluate and compare the fracture resistance of 3D-printed partial veneers with finish lines at three different locations and conventional full veneers with finish lines at the gingival level. All restorations were digitally designed and 3D printed using a nanoceramic filled resin specifically developed for veneer restorations. Methods: Four maxillary right central incisor typodont teeth were prepared for labial veneers with finish lines at different locations: incisal third (InT), middle portion of the middle third (MmT), lower portion of the middle third (LmT), and conventional veneer with the finish line at the gingival level (CoV). Each preparation was scanned, and 15 casts were 3D printed from each scan. A total of 60 3D-printed veneers were fabricated (n = 15 per group) using a nanoceramic-filled resin designed for veneer restorations. The restorations were cemented to the 3D-printed dies using the manufacturer’s adhesive and resin cement. The specimens were artificially aged with 10,000 thermal cycles between 5 °C and 55 °C, with a dwell time of 30 s, and then loaded to failure using a universal testing machine. Fracture load values were analyzed using one-way ANOVA and the Tukey honestly significant difference post hoc test (α = 0.05). In addition, fracture patterns were evaluated using scanning electron microscopy images for descriptive purposes. Results: The mean fracture resistance of the 3D-printed partial and conventional labial veneers differed significantly depending on restoration design (p < 0.05). Among the partial veneers, the LmT group showed the highest fracture resistance (279.86 N), followed by the MmT group (266.92 N), while the InT group showed the lowest value (179.22 N). The conventional veneer group (CoV) demonstrated higher fracture resistance (404.07 N) than all partial veneer groups. Conclusions: The fracture resistance of 3D-printed partial and conventional labial veneers fabricated with nanoceramic-filled resins differed according to finish line location. Conventional veneers demonstrated higher fracture resistance than all partial veneer designs. The smallest partial veneer, with the margin located in the incisal third, showed lower fracture resistance than the partial veneer designs with finish lines in the middle third. Full article