CHARACTERISTICS, OUTCOMES, AND OPTICAL COHERENCE TOMOGRAPHY FEATURES OF MACULAR HOLE AFTER RHEGMATOGENOUS RETINAL DETACHMENT REPAIR.

PURPOSE: To investigate full-thickness macular holes (FTMHs) that develop after rhegmatogenous retinal detachment (RRD) repair. METHODS: Retrospective consecutive case series of patients who underwent RRD repair between 2015 and 2021 and subsequently developed FTMH. Twenty-three patients (23 eyes) were included. All eyes underwent pars plana vitrectomy + membrane peel for macular hole (MH) repair. RESULTS: The mean age at RRD diagnosis was 62 years and 52% were male. In total, 78% were macula-off RRDs. Fourteen eyes (60.9%) underwent pars plana vitrectomy alone, 7 (30.4%) underwent pars plana vitrectomy and scleral buckling, 1 (4.3%) underwent scleral buckling only, and 1 (4.3%) underwent pneumatic retinopexy. Median time from RRD repair to MH diagnosis was 116 days. Macula-off RRDs had a significantly lower time to MH diagnosis (mean 154 days, P < 0.05) than macula-on RRDs (mean 875 days). Twenty-one MH (91%) had optical coherence tomography evidence of an epiretinal membrane. Mean logMAR visual acuity at MH diagnosis was 1.05 (∼20/224) and significantly improved to 0.66 (∼20/91) at the final follow-up ( P < 0.001); 100% of MHs closed by the final follow-up. CONCLUSION: Pars plana vitrectomy for MH after RRD repair has a high closure rate and leads to significant VA improvement. MH formation after RRD repair may be associated with macula-off detachments and epiretinal membrane. Macula-off RRDs develop MH faster than macula-on RRDs.

Biomolecules of mushroom: a recipe of human wellness.

The Indian system of medicine - Ayurveda says "When diet is wrong, medicine is of no use. When diet is correct, medicine is of no use". In this context, mushroom constitutes one of the major resources for nutraceuticals. Biomolecules of mushrooms have attracted the attention of researchers around the globe due to their proven healthy attributes. They have a plenitude of health-giving properties and these range from immunomodulatory, antiviral, antibacterial, antifungal, antioxidant, anti-inflammatory, antitumor, anticancer, anti-HIV, antidiabetic, anticholesterolic to antiarthritic activities.Mushrooms contain both primary and secondary metabolites. The primary metabolites provide energy while the secondary metabolite exhibits medicinal properties. Hence, the mushroom can be a recipe for human wellness and will play a significant role in fighting COVID-19 pandemics and other infectious diseases.The key findings suggested in this paper refer to the exploration of health and the healing traits of biomolecules of mushrooms. This article reviews the current status of the medicinal attributes of mushrooms and their biomolecules in different diseases such as cardiovascular, diabetes, reproductive diseases, cancer, and neurodegenerative diseases. The global malnutrition-related morbidity and mortality among children under five and lactating women presents a frightening picture and also a black spot on the human face. Malnutrition is responsible for more ill-health than any other cause. Mushrooms as a rich source of bioactive compounds can be claimed as "Best from the Waste" since they grow on the most abundant organic wastes of the Earth, the lignocellulosic substrate, and 'Best of the Rest' because they are excellent nutraceutical resources.

Portable Prussian Blue-Based Sensor for Bacterial Detection in Urine.

Bacterial infections can affect the skin, lungs, blood, and brain, and are among the leading causes of mortality globally. Early infection detection is critical in diagnosis and treatment but is a time- and work-consuming process taking several days, creating a hitherto unmet need to develop simple, rapid, and accurate methods for bacterial detection at the point of care. The most frequent type of bacterial infection is infection of the urinary tract. Here, we present a wireless-enabled, portable, potentiometric sensor for E. coli. E. coli was chosen as a model bacterium since it is the most common cause of urinary tract infections. The sensing principle is based on reduction of Prussian blue by the metabolic activity of the bacteria, detected by monitoring the potential of the sensor, transferring the sensor signal via Bluetooth, and recording the output on a laptop or a mobile phone. In sensing of bacteria in an artificial urine medium, E. coli was detected in ~4 h (237 ± 19 min; n = 4) and in less than 0.5 h (21 ± 7 min, n = 3) using initial E. coli concentrations of ~103 and 105 cells mL-1, respectively, which is under or on the limit for classification of a urinary tract infection. Detection of E. coli was also demonstrated in authentic urine samples with bacteria concentration as low as 104 cells mL-1, with a similar response recorded between urine samples collected from different volunteers as well as from morning and afternoon urine samples.

Heart rate variability as a marker of cardiovascular dysautonomia in post-COVID-19 syndrome using artificial intelligence.

INTRODUCTION: Cardiovascular dysautonomia comprising postural orthostatic tachycardia syndrome (POTS) and orthostatic hypotension (OH) is one of the presentations in COVID-19 recovered subjects. We aim to determine the prevalence of cardiovascular dysautonomia in post COVID-19 patients and to evaluate an Artificial Intelligence (AI) model to identify time domain heart rate variability (HRV) measures most suitable for short term ECG in these subjects. METHODS: This observational study enrolled 92 recently COVID-19 recovered subjects who underwent measurement of heart rate and blood pressure response to standing up from supine position and a 12-lead ECG recording for 60 s period during supine paced breathing. Using feature extraction, ECG features including those of HRV (RMSSD and SDNN) were obtained. An AI model was constructed with ShAP AI interpretability to determine time domain HRV features representing post COVID-19 recovered state. In addition, 120 healthy volunteers were enrolled as controls. RESULTS: Cardiovascular dysautonomia was present in 15.21% (OH:13.04%; POTS:2.17%). Patients with OH had significantly lower HRV and higher inflammatory markers. HRV (RMSSD) was significantly lower in post COVID-19 patients compared to healthy controls (13.9 ± 11.8 ms vs 19.9 ± 19.5 ms; P = 0.01) with inverse correlation between HRV and inflammatory markers. Multiple perceptron was best performing AI model with HRV(RMSSD) being the top time domain HRV feature distinguishing between COVID-19 recovered patients and healthy controls. CONCLUSION: Present study showed that cardiovascular dysautonomia is common in COVID-19 recovered subjects with a significantly lower HRV compared to healthy controls. The AI model was able to distinguish between COVID-19 recovered patients and healthy controls.

Pleurotus sajor-caju-Mediated Synthesis of Silver and Gold Nanoparticles Active against Colon Cancer Cell Lines: A New Era of Herbonanoceutics.

Herbal medicines are widely used worldwide and much appreciated because of their fewer side effects and the ability to fight diseases at the root cause. Active 'phyto' ingredients require a scientific approach and a mechanism to distribute components at the target site for better therapeutic results. Nanotechnology, on the other hand, has created new hope for cancer treatment but is still far from being proven in clinical settings. This article combines a unique approach to synthesis with the use of Pleurotus sajor-caju, followed by microwave irritation of silver and gold nanoparticles that ensures the capping of the active phyto ingredient and further enhances the effects of nanomedicine to fight colon cancer, thus opening a new era of what we call herbonanoceutics. The article also compares the characteristics and properties of silver (Au) and gold (Ag) nanoparticles synthesized by an in house developed novel microwave-assisted rapid green synthesis method. The as-prepared Ag NPs and Au NPs were compared using ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and energy dispersive spectroscopy (EDS). Our comparative study revealed that both assemblies display face-centred cubic structures (FCCs) and are nanocrystalline in nature. The advantage of the approach was that the sizes of gold and silver were identical in range with a similar distribution pattern. This has helped us to study the activity against colon cancer cell line (HCT-116) without incoherence since size plays a key role in the application. More specifically, morphological changes, cell viability, the production of reactive oxygen species (ROS) and the fragmentation of DNA have been further reported to assess better the results obtained with the two metals. Our results suggest that the newly adopted synthesis method may ensure the dual benefits from phyto ingredients which further enhances the effectiveness of advanced nanomedicine.

Molecular mechanistic insight of hepatitis B virus mediated hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a cancer which occurs in liver and severity of this cancer makes it the sixth most prevalent cancer and second leading cause of death among all cancers. The load of hepatitis-B virus (HBV) in serum is one of the important risk factors for the HCC. Several other factors also contribute to the HBV associated malignant hepatoma (HCC) i.e. HBV mutation, integration and condition of the host. Transformation of the liver to HBV-associated HCC usually accompanies long-run symptoms i.e. inflammation and cirrhosis of the liver and infective agent load could be a vigorous prognosticator for each incidence and progression of this carcinoma. One of the prominent factors i.e. HBV X supermolecule (HBx) interferes with many signal pathways that are related to the proliferation and apoptosis of hepatic cells. Besides, HBx C-terminal truncation is also responsible for HCC. Longtime HBV infection causes risk of HCC; thus most of the study related to HBV (85%) is limited to HBV endemic regions. In this review, we have outlined the molecular mechanisms that come from other than HBV endemic places which can be innovative approaches to treat HCC.

Lipid Bilayer-like Mixed Self-Assembled Monolayers with Strong Mobility and Clustering-Dependent Lectin Affinity.

Glycans at the surface of cellular membranes modulate biological activity via multivalent association with extracellular messengers. The lack of tuneable simplified models mimicking this dynamic environment complicates basic studies of these phenomena. We here present a series of mixed reversible self-assembled monolayers (rSAMs) that addresses this deficiency. Mixed rSAMs were prepared in water by simple immersion of a negatively charged surface in a mixture of sialic acid- and hydroxy-terminated benzamidine amphiphiles. Surface compositions derived from infrared reflection-absorption spectroscopy (IRAS) and film thickness information (atomic force microscopy, ellipsometry) suggest the latter to be statistically incorporated in the monolayer. These surfaces' affinity for the lectin hemagglutinin revealed a strong dependence of the affinity on the presentation, density, and mobility of the sialic acid ligands. Hence, a spacer length of 4 ethylene glycol and a surface density of 15% resulted in a dissociation constant Kd,multi of 1.3 × 10-13 M, on par with the best di- or tri-saccharide-based binders reported to date, whereas a density of 20% demonstrated complete resistance to hemagglutinin binding. These results correlated with ligand mobility measured by fluorescence recovery after photobleaching which showed a dramatic drop in the same interval. The results have a direct bearing on biological cell surface multivalent recognition involving lipid bilayers and may guide the design of model surfaces and sensors for both fundamental and applied studies.

Accuracy of left atrial bipolar voltages obtained by ConfiDENSE multielectrode mapping in patients with persistent atrial fibrillation.

INTRODUCTION: The ConfiDENSE™ module (Carto3 v4) allows rapid annotation of endocardial electrograms acquired by multielectrode (ME) mapping. However, its accuracy in assessing atrial voltages is unknown. METHODS AND RESULTS: Two ConfiDENSE™ left atrial voltage maps were created during continuous pacing in 20 patients undergoing catheter ablation for persistent AF using a ME lasso catheter and a contact force (CF) sensing ablation catheter. The automated tissue proximity indicator (TPI) filter was then applied to the ME map to yield a TPI map. Reference maps (RM) were created offline by a blinded observer by manually assessing all points against fidelity criteria. Bipolar voltages and proportion of low voltage points (< 0.5 mV) derived from the ME, CF, and TPI maps were compared with those derived from the RM. Note that 853 ± 365 points, 252 ± 184 points, and 144 ± 73 were collected for ME, TPI, and CF maps, respectively, and 429 ± 153 points were included in the RM. Voltages with CF and TPI maps were similar to those with RM (1.57 ± 0.47 mV vs. 1.63 ± 0.31 mV, P = 0.57 and 1.50 ± 0.38 mV vs. 1.63 ± 0.31 mV, P = 0.07, respectively), whereas ME maps showed a significantly lower mean voltage (1.00 ± 0.22 mV, P < 0.001). As compared to RM maps (17 ± 8%), low voltage points were significantly overestimated by the ME maps (50 ± 9% (P < 0.001) and TPI maps (28 ± 13% (P < 0.001), but not by the CF maps (22 ± 14%, P = 0.17). CONCLUSION: Application of the TPI filter to ConfiDENSE maps significantly increases the quality of the voltage data, conserving a reasonable point density, but still overestimates low voltage points as compared to CF-sensing maps or maps reviewed manually.

Prospective use of Ablation Index targets improves clinical outcomes following ablation for atrial fibrillation.

AIMS: Late recovery of ablated tissue leading to reconnection of pulmonary veins remains common following radiofrequency catheter ablation for AF. Ablation Index (AI), a novel ablation quality marker, incorporates contact force (CF), time, and power in a weighted formula. We hypothesized that prospective use of our previously published derived AI targets would result in better outcomes when compared to CF-guided ablation. METHODS: Eighty-nine consecutive drug-refractory AF patients (49% paroxysmal) underwent AI-guided ablation (AI-group). AI targets were 550 for anterior/roof and 400 for posterior/inferior left atrial segments. Procedural and clinical outcomes of these patients were compared to 89 propensity-matched controls who underwent CF-guided ablation (CF-group). All 178 procedures were otherwise similar, and both groups were followed-up for 12 months. The last 25 patients from each group underwent analysis of all VisiTags™ for ablation duration, CF, Force-Time Integral, and impedance drop. RESULTS: First-pass pulmonary vein isolation (PVI) was more frequent in AI-group than in CF-group (173 [97%] vs. 149 [84%] circles, P < 0.001), and acute PV reconnection was lower (11 [6%] vs. 24 [13%] circles, P = 0.02). Mean PVI ablation time was similar (AI-group: 42 ± 9 vs. CF-group: 45 ± 14 minutes, P = 0.14). Median impedance drop for AI-group was significantly higher than in CF-group (13.7 [9-19] Ω vs. 8.8 [5.2-13] Ω, P < 0.001). Two major complications occurred in CF-group and none in AI-group. Atrial tachyarrhythmia recurrence was significantly lower in AI-group (15 of 89 [17%]) than in CF-group (33 of 89 [37%], P = 0.002). CONCLUSION: AI-guided ablation is associated with significant improvements in the incidence of acute PV reconnection and atrial tachyarrhythmia recurrence rate compared to CF-guided ablation, potentially due to creation of better quality lesions as suggested by greater impedance drop.

Double orifice mitral valve in rheumatic heart disease.

A 55-year-old female, presented to our outpatient department with complaints of dyspnea on exertion, NYHA grade II for 7 years, which had progressed to NYHA Grade III in the past 6 months. An echocardiogram done showed severe mitral stenosis with mitral valve fused in the middle to create an appearance of 2 separate valves (complete bridge type) with 2 separate turbulent jets of flow across these valves. No other congenital anomaly was seen. Due to presence of severe subvalvular disease, she was sent for elective mitral valve replacement. This case demonstrates that a careful analysis of the subvalvular apparatus of the mitral valve is needed before making a decision for definitive treatment of this condition.

Intermittent Right Ventricular Outflow Tract Capture due to Chronic Right Atrial Lead Dislodgement.

A 58 year old male, known case of type 2 diabetes and hypertension, had undergone implantation of a dual chamber pacemaker(DDDR) in 2007 for complaints of recurrent syncope and trifascicular block with a normal ejection fraction andnormal coronaries. His post implantation parameters were normal at that time.He now presented to our pacemaker clinic where his ECG done showed two types o fpaced complexes. The first few complexes were consistent with atrial sensed right ventricular apical pacing with left superior axis. Later complexes showed loss of atrial sensing with pacing from right ventricular outflow tract(inferior axis) with subtle oscillation in it's axis. On application of magnet, two pacemaker spikes were visible withinterspike interval of 120 ms and paced complexes with inferior axis starting from the first spike suggesting that the atrial lead was responsible for RVOT depolarization. On interrogation of the pacemaker, atrial EGM showed sensed activity from atrium followed by large sensed ventricular complex. Fluoroscopy confirmed that the atrial lead was dislodged and was intermittently prolapsing into the RVOT. Since the patient was asymptomatic, he refused any intervention and subsequentlyhis atrial lead was switched off by telemetry. The above case signifies that asymptomatic lead dislodgement is no talways manifested as loss of capture and even subtle variation of the axis o fthe paced complexes can provide us with a clue that can be confirmed by telemetry of the pacemaker and fluoroscopy.

Scleral Buckle Removal: Long-Term Patient Outcomes.

PURPOSE: Scleral buckling has been a reliable treatment option in the repair of primary rhegmatogenous retinal detachments. Occasionally, patients require scleral buckles (SBs) to be removed for various reasons. While outcomes of SB removal have been investigated in this subset of patients, there has not been any large patient series to reach any conclusions. Long-term sequelae of SB removal are debated in the literature, specifically around the risk of redetachment. DESIGN: We performed a retrospective, observational study to evaluate the clinical indications for, and outcomes of, SB removal. PARTICIPANTS: No control patients in this retrospective, observational study. METHODS: Eighty-six individuals with a history of SB removal from June 1, 2000, to January 1, 2021, were followed from a large academic center and a private, retina-only practice in Chicago. Exclusion criteria were age of < 18 years and unplanned or self-explanted SB removal. MAIN OUTCOME MEASURES: Data extracted included patient symptoms before SB removal, indications for removal, resolution of symptoms following removal, rate of redetachment, and rate of additional ocular surgery. Secondary outcomes included identifying factors associated with poorer outcomes. RESULTS: Eighty-six eyes with history of SB removal were included with an average follow-up of 4 years. Approximately 60% were males and the mean age at the time of SB removal was 59 years. Leading indications for removal were exposure (61.63%), infection (20.93%), and diplopia/strabismus (19.77%). The average time from SB placement to removal was 12.28 ± 11.16 years. Most patients requiring SB removal presented with symptoms, specifically of pain and discomfort (65.12%), diplopia (22.09%), and drainage/discharge (18.60%). Of these patients, 86.59% experienced symptom resolution following SB removal. Notably, 6.56% (4 eyes) of all eyes with at least 1 year of follow-up experienced a redetachment requiring surgery. Within this subset, the average time from SB placement to removal was 2.05 ± 2.01 years and time to redetachment following removal was 15.95 ± 25.71 months. Nine percent of all eyes required additional strabismus or oculoplastic surgery. CONCLUSIONS: Scleral buckle removal provides a high rate of symptomatic relief and low risk of subsequent detachment. Nevertheless, close monitoring is warranted to monitor for recurrent retinal detachments. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Folate-Mediated Targeting and Controlled Release: PLGA-Encapsulated Mesoporous Silica Nanoparticles Delivering Capecitabine to Pancreatic Tumor.

The discovery of specifically tailored therapeutic delivery systems has sparked the interest of pharmaceutical researchers considering improved therapeutic effectiveness and fewer adverse effects. The current study concentrates on the design and characterization of PLGA (polylactic-co-glycolic acid) capped mesoporous silica nanoparticles (MSN)-based systems for drug delivery for pH-sensitive controlled drug release in order to achieve a targeted drug release inside the acidic tumor microenvironment. The physicochemical properties of the nanoformulations were analyzed using TEM, zeta potential, AFM, TGA, FTIR, and BET analyses in addition to DLS size. The final formed PLGA-FoA-MSN-CAP and pure MSN had sizes within the therapeutic ranges of 164.5 ± 1.8 and 110.7 ± 2.2, respectively. Morphological characterization (TEM and AFM) and elemental analysis (FTIR and XPS) confirmed the proper capping and tagging of PLGA and folic acid (FoA). The PLGA-coated FoA-MSN exhibited a pH-dependent controlled release of the CAP (capecitabine) drug, showing efficient release at pH 6.8. Furthermore, the in vitro MTT test on PANC1 and MIAPaCa-2 resulted in an IC50 value of 146.37 µg/ml and 105.90 µg/ml, respectively. Mitochondrial-mediated apoptosis was confirmed from the caspase-3 and annexin V/PI flow cytometry assay, which displayed a cell cycle arrest at the G1 phase. Overall, the results predicted that the designed nanoformulation is a potential therapeutic agent in treating pancreatic cancer.

Pseudo-aneurysm of mitral aortic intervalvular fibrosa: two case reports.

The fibrous body between the mitral and aortic valve, known as mitral-aortic intervalvular fibrosa (MAIVF) is prone to infection and injury resulting in pseudo-aneurysm formation. Because of its relative rarity, we are far from making any conclusion regarding the natural history and appropriate therapeutic strategy for this condition. We report two cases of this condition with two different and rare etiologies with strikingly different natural courses, providing insight into the natural course and timing of surgery in this rare entity.

A Facile Synthesis of Flower-like Iron Oxide Nanoparticles and Its Efficacy Measurements for Antibacterial, Cytotoxicity and Antioxidant Activity.

The objective of this study was to investigate the rhombohedral-structured, flower-like iron oxide (Fe2O3) nanoparticles that were produced using a cost-effective and environmentally friendly coprecipitation process. The structural and morphological characteristics of the synthesized Fe2O3 nanoparticles were analyzed using XRD, UV-Vis, FTIR, SEM, EDX, TEM, and HR-TEM techniques. Furthermore, the cytotoxic effects of Fe2O3 nanoparticles on MCF-7 and HEK-293 cells were evaluated using in vitro cell viability assays, while the antibacterial activity of the nanoparticles against Gram-positive and Gram-negative bacteria (Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae) was also tested. The results of our study demonstrated the potential cytotoxic activity of Fe2O3 nanoparticles toward MCF-7 and HEK-293 cell lines. The antioxidant potential of Fe2O3 nanoparticles was evidenced by the 1,1-diphenyl-2-picrylhydrazine (DPPH) and nitric oxide (NO) free radical scavenging assays. In addition, we suggested that Fe2O3 nanoparticles could be used in various antibacterial applications to prevent the spread of different bacterial strains. Based on these findings, we concluded that Fe2O3 nanoparticles have great potential for use in pharmaceutical and biological applications. The effective biocatalytic activity of Fe2O3 nanoparticles recommends its use as one of the best drug treatments for future views against cancer cells, and it is, therefore, recommended for both in vitro and in vivo in the biomedical field.

Biosynthesized nanoparticles: a novel approach for cancer therapeutics.

Nanotechnology has become one of the most rapid, innovative, and adaptable sciences in modern science and cancer therapy. Traditional chemotherapy has limits owing to its non-specific nature and adverse side effects on healthy cells, and it remains a serious worldwide health issue. Because of their capacity to specifically target cancer cells and deliver therapeutic chemicals directly to them, nanoparticles have emerged as a viable strategy for cancer therapies. Nanomaterials disclose novel properties based on size, distribution, and shape. Biosynthesized or biogenic nanoparticles are a novel technique with anti-cancer capabilities, such as triggering apoptosis in cancer cells and slowing tumour growth. They may be configured to deliver medications or other therapies to specific cancer cells or tumour markers. Despite their potential, biosynthesized nanoparticles confront development obstacles such as a lack of standardisation in their synthesis and characterization, the possibility of toxicity, and their efficiency against various forms of cancer. The effectiveness and safety of biosynthesized nanoparticles must be further investigated, as well as the types of cancer they are most successful against. This review discusses the promise of biosynthesized nanoparticles as a novel approach for cancer therapeutics, as well as their mode of action and present barriers to their development.

Computational Approaches to Designing Antiviral Drugs against COVID-19: A Comprehensive Review.

The global impact of the COVID-19 pandemic caused by SARS-CoV-2 necessitates innovative strategies for the rapid development of effective treatments. Computational methodologies, such as molecular modelling, molecular dynamics simulations, and artificial intelligence, have emerged as indispensable tools in the drug discovery process. This review aimed to provide a comprehensive overview of these computational approaches and their application in the design of antiviral agents for COVID-19. Starting with an examination of ligand-based and structure-based drug discovery, the review has delved into the intricate ways through which molecular modelling can accelerate the identification of potential therapies. Additionally, the investigation extends to phytochemicals sourced from nature, which have shown promise as potential antiviral agents. Noteworthy compounds, including gallic acid, naringin, hesperidin, Tinospora cordifolia, curcumin, nimbin, azadironic acid, nimbionone, nimbionol, and nimocinol, have exhibited high affinity for COVID-19 Mpro and favourable binding energy profiles compared to current drugs. Although these compounds hold potential, their further validation through in vitro and in vivo experimentation is imperative. Throughout this exploration, the review has emphasized the pivotal role of computational biologists, bioinformaticians, and biotechnologists in driving rapid advancements in clinical research and therapeutic development. By combining state-of-the-art computational techniques with insights from structural and molecular biology, the search for potent antiviral agents has been accelerated. The collaboration between these disciplines holds immense promise in addressing the transmissibility and virulence of SARS-CoV-2.