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BackgroundIn this study, the preliminary experience of endoscopic nipple-areolar-complex (NAC) sparing total mastectomy were analyzed and reported.MethodsThe medical records of the patients who underwent Endoscopic NAC sparing total mastectomy from November 2019 to June 2022 in a single institute were collected and analyzed. The medical records of their clinicopathologic characteristics, perioperative parameters, postoperative complications, oncologic results were collected retrospectively. The learning curve was evaluated using univariable and multivariable analyses and analyzed using the CUSUM method.ResultsA total of 56 surgeries from 53 patients were analyzed. The mean total operation time was 272.50±66.74 minutes while mean oncologic mastectomy was 155.61±43.21 minutes. The mean postoperative hospital days were 9.85±4.70 days. In CUSUM analysis, the cases needed to decrease operation time were 10th. The overall complication rate related to implant was 15.09%. There were one locoregional recurrence patient and one distant metastasis and mortality patient observed during median follow-up of 19.12±12.01 months.ConclusionAccording to our preliminary study, endoscopic NAC sparing total mastectomy is a safe procedure, and the operation time can be reduced with accumulated surgical experience.

The ribosomal genes (rDNA genes) encode 47S rRNA which accounts for up to 80% of all cellular RNA. At any given time, no more than 50% of rDNA genes are actively transcribed, and the other half is silent by forming heterochromatin structures through DNA methylation. In cancer cells, upregulation of ribosome biogenesis has been recognized as a hallmark feature, thus, the reduced methylation of rDNA promoter has been thought to support conformational changes of chromatin accessibility and the subsequent increase in rDNA transcription. However, an increase in the heterochromatin state through rDNA hypermethylation can be a protective mechanism teetering on the brink of a threshold where cancer cells rarely successfully proliferate. Hence, clarifying hypo- or hypermethylation of rDNA will unravel its additional cellular functions, including organization of genome architecture and regulation of gene expression, in response to growth signaling, cellular stressors, and carcinogenesis. Using the bisulfite-based quantitative real-time methylation-specific PCR (qMSP) method after ensuring unbiased amplification and complete bisulfite conversion of the minuscule DNA amount of 1 ng, we established that the rDNA promoter was significantly hypermethylated in 107 breast, 65 lung, and 135 colon tumour tissue samples (46.81%, 51.02% and 96.60%, respectively) as compared with their corresponding adjacent normal samples (26.84%, 38.26% and 77.52%, respectively; p < 0.0001). An excessive DNA input of 1 μg resulted in double-stranded rDNA remaining unconverted even after bisulfite conversion, hence the dramatic drop in the single-stranded DNA that strictly required for bisulfite conversion, and leading to an underestimation of rDNA promoter methylation, in other words, a faulty hypomethylation status of the rDNA promoter. Our results are in line with the hypothesis that an increase in rDNA methylation is a natural pathway protecting rDNA repeats that are extremely sensitive to DNA damage in cancer cells.

[This corrects the article DOI: 10.1371/journal.pone.0294647.].

This study investigates the impact of arc length, current intensity, travel speed, and gas flow rate on surface hardness and line width during arc quenching process of S45C steel. The current intensity has the greatest influence on the surface hardness of S45C steel, followed by the travel speed, gas flow rate, and arc length. Using the Taguchi method, the optimal values of the parameters such as the arc length of 1.5 mm, the current intensity of 125 A, the travel speed of 250 mm/min and the gas flow rate of 12.5 l/min were calculated. The optimal surface hardness would be 379 HV, with a standard deviation of 46.4 HV. The current intensity is the most critical component in determining line width among these parameters. The arc length ranks second, followed by the TIG gun's travel speed. The gas flow rate is the least significant factor. A longer arc length may result in a broader heat zone, which leads to a better line width. Increasing the arc length, current intensity, travel speed, and gas flow rate results in a similar pattern of surface hardness change caused by the low-heated and over-heated phenomena. The microhardness distribution showed a hardening zone of up to 2500 μm and a maximum hardness of 453 HV. The microstructure of arc quenching samples has three zones: hardening, heat-affected, and base metal. The hardening zone exhibits a martensite microstructure with a tiny needle shape and a residual austenite matrix.

This study investigates the impact of arc length, current intensity, travel speed, and gas flow rate on surface hardness and line width during arc quenching process of S45C steel. The current intensity has the greatest influence on the surface hardness of S45C steel, followed by the travel speed, gas flow rate, and arc length. Using the Taguchi method, the optimal values of the parameters such as the arc length of 1.5 mm, the current intensity of 125 A, the travel speed of 250 mm/min and the gas flow rate of 12.5 l/min were calculated. The optimal surface hardness would be 379 HV, with a standard deviation of 46.4 HV. The current intensity is the most critical component in determining line width among these parameters. The arc length ranks second, followed by the TIG gun's travel speed. The gas flow rate is the least significant factor. A longer arc length may result in a broader heat zone, which leads to a better line width. Increasing the arc length, current intensity, travel speed, and gas flow rate results in a similar pattern of surface hardness change caused by the low-heated and over-heated phenomena. The microhardness distribution showed a hardening zone of up to 2500 μm and a maximum hardness of 453 HV. The microstructure of arc quenching samples has three zones: hardening, heat-affected, and base metal. The hardening zone exhibits a martensite microstructure with a tiny needle shape and a residual austenite matrix. [ABSTRACT FROM AUTHOR]

The autonomous and active Long-Interspersed Element-1 (LINE-1, L1) and the non-autonomous Alu retrotransposon elements, contributing to 30% of the human genome, are the most abundant repeated sequences. With more than 90% of their sequences being methylated in normal cells, these elements undeniably contribute to the global DNA methylation level and constitute a major part of circulating-cell-free DNA (cfDNA). So far, the hypomethylation status of LINE-1 and Alu in cellular and extracellular DNA has long been considered a prevailing hallmark of ageing-related diseases and cancer. This study demonstrated that errors in LINE-1 and Alu methylation level measurements were caused by an excessive input quantity of genomic DNA used for bisulfite conversion. Using the minuscule DNA amount of 0.5 ng, much less than what has been used and recommended so far (500 ng-2 μg) or 1 μL of cfDNA extracted from 1 mL of blood, we revealed hypermethylation of LINE-1 and Alu in 407 tumour samples of primary breast, colon and lung cancers when compared with the corresponding pair-matched adjacent normal tissue samples (P < 0.05-0.001), and in cfDNA from 296 samples of lung cancers as compared with 477 samples from healthy controls (P < 0.0001). More importantly, LINE-1 hypermethylation in cfDNA is associated with healthy ageing. Our results have not only contributed to the standardized bisulfite-based protocols for DNA methylation assays, particularly in applications on repeated sequences but also provided another perspective for other repetitive sequences whose epigenetic properties may have crucial impacts on genome architecture and human health.

This paper investigates the performance of hybrid radio frequency/free space optical (RF/FSO) systems combined with non-orthogonal multiple access communications technology. We examine a scenario where the source and destination are separated by a large distance, with no direct link between them. The relay, denoted R, operates using the decode-and-forward (DF) protocol. Under the DF relaying scheme, the relay employs successive interference cancellation (SIC). In this setup, the FSO link from the source to the relay follows a Gamma-Gamma distribution, while the RF link from the relay to multiple users follow a Nakagami-m distribution. Based on this system model, we analyze the outage probability (OP). Our findings indicate a direct relationship between SIC and OP performance: the higher the SIC capability, the more effective the system. In addition, the system's performance is dependent on the parameters of the FSO channel. Finally, Monte Carlo simulations are presented to further validate our framework and findings.

[This corrects the article DOI: 10.1371/journal.pone.0257449.].

It is important to establish the relationship between appropriate nutritional intake and improvements in activities of daily living (ADLs) in elderly hospitalized patients. This prospective observational study aimed to investigate diet order compliance (DOC) during 8 weeks of hospitalization and calculate the odds of improved functional independence measure (FIM) scores for high- and low-DOC groups using covariate-adjusted models in geriatric convalescent hospitals. The study subjects were elderly inpatients (>65 years old) with degenerative disease who consumed prescribed oral meals (Functional Oral Intake Scale (FOIS) = 6/7) and who did not receive physical/occupational therapy. The personalized diet order was prescript, and the DOC was calculated using dietitian-monitored daily intake data. The 73 patients were divided into a low-DOC group (< 84.0%, n = 35) and a high-DOC group (≥ 84.0%, n = 38) on the basis of the median DOC (84.0%, average for 8 weeks). Twenty (52.6%) high-DOC patients and nine (25.7%) low-DOC patients experienced motor-FIM improvements (P = 0.017). After 8 weeks, the change in motor-FIM in the high-DOC group (1.6±0.3) was greater than that in the low-DOC group (0.3±0.1; P = 0.001). According to the baseline and nutrition-intake-adjusted model of multiple logistic regression analysis, in the high-DOC group, the motor-FIM improvement OR was 5.102 (95% CI: 1.100-16.233, P = 0.036), and the total-FIM improvement OR was 5.273 (95% CI: 1.102-25.238, P = 0.037). High compliance with individualized nutritional prescriptions can increase FIM scores in clinical settings. Thus, comprehensive approaches to increase dietary compliance are needed for elderly long-term care patients.

[This corrects the article DOI: 10.1371/journal.pone.0270808.].

BackgroundHypertension (HTN) is linked to an enhanced risk of chronic kidney disease and cardiovascular disease. While sodium and potassium intake affect blood pressure (BP) control, the urine sodium-to-potassium (Na/K) ratio, which reflects dietary balance and renal regulation of these electrolytes, could be associated with BP. This study aimed to evaluate the independent association between urine Na/K and uncontrolled HTN.MethodsData were collected from the Korea National Health and Nutrition Examination Survey from 2016 to 2021. A total of 5,770 participants diagnosed with HTN were enrolled in this study. Uncontrolled HTN was characterized by a systolic blood pressure (SBP) ≥ 140 mmHg or diastolic blood pressure (DBP) ≥ 90 mmHg. Logistic regression analysis was used to assess the relationship between urine Na/K and the risk of uncontrolled HTN.ResultsThe urine Na/K was positively correlated with both SBP and mean arterial pressure. Higher urine Na/K was significantly associated with an increased risk of uncontrolled HTN using both continuous (odds ratio [95% confidence interval] 1.13 [1.09-1.16], P ConclusionUrine Na/K ratio is independently associated with uncontrolled HTN in the general population and in patients with CKD. Our findings suggest that monitoring the urine Na/K could serve as an effective tool for identifying subjects at risk of uncontrolled HTN.

[This corrects the article DOI: 10.1371/journal.pone.0303758.].

Although sensor technology has advanced with better materials, biomarkers, and fabrication and detection methods, creating a rapid, accurate, and affordable bacterial detection platform is still a major challenge. In this study, we present a combination of hybrid-MoS2 nanosheets and an amine-customized probe to develop a fast, sensitive biosensor for Bacillus subtilis DNA detection. Based on fluorescence measurements, the biosensor exhibits a detection range of 23.6-130 aM, achieves a detection limit of 18.7 aM, and was stable over four weeks. In addition, the high selectivity over Escherichia coli and Vibrio proteolyticus DNAs of the proposed Bacillus subtilis sensors is demonstrated by the fluorescence quenching effect at 558 nm. This research not only presents a powerful tool for B. subtilis DNA detection but also significantly contributes to the advancement of hybrid 2D nanomaterial-based biosensors, offering substantial promise for diverse applications in biomedical research and environmental monitoring.

There currently exists no comprehensive and up-to date overview on the financial impact of the different adverse events covered by the Patient Safety Indicators (PSIs) from the Agency for Healthcare Research and Quality. We conducted a retrospective case-control study using propensity score matching on a national administrative data set of 1 million inpatients in Switzerland to compare excess costs associated with 16 different adverse events both individually and on a nationally aggregated level. After matching 8,986 cases with adverse events across the investigated PSIs to 26,931 controls, we used regression analyses to determine the excess costs associated with the adverse events and to control for other cost-related influences. The average excess costs associated with the PSI-related adverse events ranged from CHF 1,211 (PSI 18, obstetric trauma with instrument) to CHF 137,967 (PSI 10, postoperative acute kidney injuries) with an average of CHF 27,409 across all PSIs. In addition, adverse events were associated with 7.8-day longer stays, 2.5 times more early readmissions (within 18 days), and 4.1 times higher mortality rates on average. At a national level, the PSIs were associated with CHF 347 million higher inpatient costs in 2019, which corresponds to about 2.2% of the annual inpatient costs in Switzerland. By comparing the excess costs of different PSIs on a nationally aggregated level, we offer a financial perspective on the implications of in-hospital adverse events and provide recommendations for policymakers regarding specific investments in patient safety to reduce costs and suffering.

Diabetes mellitus is characterized by insulin resistance and β-cell failure. The latter involves impaired insulin secretion and β-cell dedifferentiation. Sulfonylurea (SU) is used to improve insulin secretion in diabetes, but it suffers from secondary failure. The relationship between SU secondary failure and β-cell dedifferentiation has not been examined. Using a model of SU secondary failure, we have previously shown that functional loss of oxidoreductase Cyb5r3 mediates effects of SU failure through interactions with glucokinase. Here we demonstrate that SU failure is associated with partial β-cell dedifferentiation. Cyb5r3 knockout mice show more pronounced β-cell dedifferentiation and glucose intolerance after chronic SU administration, high-fat diet feeding, and during aging. A Cyb5r3 activator improves impaired insulin secretion caused by chronic SU treatment, but not β-cell dedifferentiation. We conclude that chronic SU administration affects progression of β-cell dedifferentiation and that Cyb5r3 activation reverses secondary failure to SU without restoring β-cell dedifferentiation.

[This corrects the article DOI: 10.1371/journal.pone.0210248.].

The rapid and accurate detection of pathogenic bacteria is essential for food safety and public health. Conventional detection techniques, such as nucleic acid sequence-based amplification and polymerase chain reaction, are time-consuming and require specialized equipment and trained personnel. Here, we present quick, disposable impedance sensors based on the novel hybrid MoS2 nanomaterial for detecting Escherichia coli DNA. Our results indicate that the proposed sensors operate linearly between 10- 20 and 10-15 M concentrations, achieving an impressive detection limit of 10-20 M with the highest sensitivity observed at a 0.325 nM probe concentration sensor. Furthermore, the electrochemical impedance spectroscopy biosensors exhibited potential selectivity for Escherichia coli DNA over Bacillus subtilis and Vibrio proteolyticus DNA sequences. The findings offer a promising avenue for efficient and precise DNA detection, with potential implications for broader biotechnology and medical diagnostics applications.

Stroke is a leading cause of death and long-term disability which can cause oxidative damage and inflammation of the neuronal cells. Retinoic acid is an active metabolite of vitamin A that has various beneficial effects including antioxidant and anti-inflammatory effects. In this study, we investigated whether retinoic acid modulates oxidative stress and inflammatory factors in a stroke animal model. A middle cerebral artery occlusion (MCAO) was performed on adult male rats to induce focal cerebral ischemia. Retinoic acid (5 mg/kg) or vehicle was injected into the peritoneal cavity for four days before MCAO surgery. The neurobehavioral tests were carried out 24 h after MCAO and cerebral cortex tissues were collected. The cortical damage was assessed by hematoxylin-eosin staining and reactive oxygen species assay. In addition, Western blot and immunohistochemical staining were performed to investigate the activation of glial cells and inflammatory cytokines in MCAO animals. Ionized calcium-binding adapter molecule-1 (Iba-1) and glial fibrillary acidic protein (GFAP) were used as markers of microglial and astrocyte activation, respectively. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were used as representative pro-inflammatory cytokines. Results showed that MCAO damage caused neurobehavioral defects and histopathological changes in the ischemic region and increased oxidative stress. Retinoic acid treatment reduced these changes caused by MCAO damage. We detected increases in Iba-1 and GFAP in MCAO animals treated with vehicle. However, retinoic acid alleviated increases in Iba-1 and GFAP caused by MCAO damage. Moreover, MCAO increased levels of nuclear factor-κB and pro-inflammatory cytokines, including TNF-α and IL-1β. Retinoic acid alleviated the expression of these inflammatory proteins. These findings elucidate that retinoic acid regulates microglia and astrocyte activation and modulates pro-inflammatory cytokines. Therefore, this study suggests that retinoic acid exhibits strong antioxidant and anti-inflammatory properties by reducing oxidative stress, inhibiting neuroglia cell activation, and preventing the increase of pro-inflammatory cytokines in a cerebral ischemia.

BackgroundPrevious studies have reported inconsistent results regarding the advantages or disadvantages of spironolactone use in patients undergoing hemodialysis (HD). This study aimed to evaluate survival according to the use of spironolactone in a large sample of patients undergoing maintenance HD.MethodsThis retrospective study used laboratory and clinical data from the national HD Quality Assessment Program and claims data. The participants of the quality assessment program were patients who had been undergoing maintenance HD for ≥ 3 months, patients undergoing HD at least twice a week. Patients with no spironolactone prescription during the assessment periods were designated as the control group. Patients with one or more prescriptions of spironolactone during the assessment periods were assigned to the SPR group.ResultsThe number of patients in the control and SPR groups were 54,588 and 315, respectively. The 5-year survival rates were 69.1% and 59.1% in the control and SPR groups, respectively (P < 0.001). Cox regression analyses showed that the hazard ratio in the SPR group was 1.34 (P < 0.001) in univariate analysis and 1.13 (P = 0.249) in multivariable analysis. Univariate Cox-regression analysis showed a better patient survival rate in the control group than in the SPR group; however, multivariable analyses showed similar patient survival rates between the two groups.ConclusionThis study showed no difference in survival between patients undergoing HD with and without spironolactone use.

This study aimed to investigate the efficacy and safety of using optimized parameters obtained by computer simulation for ultrasound-guided high-intensity focused ultrasound (HIFU) treatment of uterine adenomyosis in comparison with conventional parameters. We retrospectively assessed a single-institution, prospective study that was registered at Clinical Research Information Service (CRiS) of Republic of Korea (KCT0003586). Sixty-six female participants (median age: 44 years) with focal uterine adenomyosis were prospectively enrolled. All participants were treated with a HIFU system by using treatment parameters either for treating uterine fibroids (Group A, first 20 participants) or obtained via computer simulation (Group B, later 46 participants). To assess the treatment efficacy of HIFU, qualitative indices, including the clinically effective dysmenorrhea improvement index (DII), were evaluated up to 3 years after treatment, whereas quantitative indices, such as the nonperfused volume ratio and adenomyosis volume shrinkage ratio (AVSR), on MRI were evaluated up to 3 months after treatment. Quantitative/qualitative indices were compared between Groups A and B by using generalized linear mixed effect model. A safety assessment was also performed. Results showed that clinically effective DII was more frequently observed in Group B than in Group A (odds ratio, 3.69; P = 0.025), and AVSR were higher in Group B than in Group A (least-squares means, 21.61; P = 0.001). However, two participants in Group B developed skin burns at the buttock and sciatic nerve pain and required treatment. In conclusion, parameters obtained by computer simulation were more effective than the conventional parameters for treating uterine adenomyosis by using HIFU in terms of clinically effective DII and AVSR. However, care should be taken because of the risk of adverse events.

[This corrects the article DOI: 10.1371/journal.pone.0017830.].

Emerging technologies focused on the detection and quantification of circulating tumor DNA (ctDNA) in blood show extensive potential for managing patient treatment decisions, informing risk of recurrence, and predicting response to therapy. Currently available tissue-informed approaches are often limited by the need for additional sequencing of normal tissue or peripheral mononuclear cells to identify non-tumor-derived alterations while tissue-naïve approaches are often limited in sensitivity. Here we present the analytical validation for a novel ctDNA monitoring assay, FoundationOne®Tracker. The assay utilizes somatic alterations from comprehensive genomic profiling (CGP) of tumor tissue. A novel algorithm identifies monitorable alterations with a high probability of being somatic and computationally filters non-tumor-derived alterations such as germline or clonal hematopoiesis variants without the need for sequencing of additional samples. Monitorable alterations identified from tissue CGP are then quantified in blood using a multiplex polymerase chain reaction assay based on the validated SignateraTM assay. The analytical specificity of the plasma workflow is shown to be 99.6% at the sample level. Analytical sensitivity is shown to be >97.3% at ≥5 mean tumor molecules per mL of plasma (MTM/mL) when tested with the most conservative configuration using only two monitorable alterations. The assay also demonstrates high analytical accuracy when compared to liquid biopsy-based CGP as well as high qualitative (measured 100% PPA) and quantitative precision (

ObjectiveTo investigate the correlation between oxygen saturation index (OSI) and oxygenation index (OI) for evaluating the blood oxygenation status in neonates with respiratory failure requiring mechanical ventilation support and to assess the predictive capability of OSI in determining clinically relevant OI cutoffs.MethodsA prospective study was conducted on neonates who received invasive mechanical ventilation at the neonatal intensive care unit of tertiary hospital in Vietnam. Bland-Altman analysis was utilized to evaluate the agreement between OSI and OI.ResultsA total of 123 neonates, including both term and preterm infants, were included in the study. A high agreement rate of 94.3% within the 95% limits of agreement (between OI and OSI), with a narrow similarity value of 3.3 (95% CI: -5.1 to 11.8) and high correlation coefficient (r = 0.791, p15 was determined to be 7.45, with a sensitivity of 100% and a specificity of 87.4% (AUC 0.955; 95% CI: 0.922-0.989, p < 0.05). Similarly, an OSI cutoff value of 9.9 corresponded to an OI of 25, displaying a sensitivity of 100% and specificity of 87.4% (AUC 0.92). The receiver operating characteristic (ROC) curves for OSI exhibited statistically significant results (p < 0.05).ConclusionThe findings demonstrate a strong correlation between OSI and OI in neonates with respiratory failure. Furthermore, OSI, as a non-invasive method, can serve as a substitute for OI to evaluate the severity of hypoxic respiratory failure and lung injury in neonates.

Tidal flats provide critical habitat for migratory waterbird species; however, populations of migratory waterbirds have significantly declined due to tidal flat loss and degradation caused by human activities, particularly in Asia. Gochang getbol is one of tidal flats located on the southwest coast of South Korea and a center of clam production. Using bird monitoring data collected at five zones (zone1 to zone5) established across Gochang getbol and near coastal area, we examined distribution patterns of migratory bird diversity and conservation-related species along the coast of Gochang getbol. The intensity of human activity ‒ mudflat culture (mostly bivalve) and aquaculture was relatively high at zone2 and zone3, occupying > 30% of 2km circular area surrounding most sample points of these zones. Zone1 and particularly zone4 contained more natural/semi-natural habitats (less disturbed mudflats and wetlands) and zone5 had smallest mudflat than others. Shannon diversity, species richness, and abundance of migratory birds differed between zones (Anova test, P ≤ 0.02) except Shannon diversity in winter. In fall, all values were higher at zone4 than zone3 and zone5. In winter, zone1 showed greatest species richness and higher abundance than zone2, zone3, and zone5. In spring, while most differences were found between zone4 and zone5, abundance at zone4 was somewhat higher than zone2. The results from the fourth corner analysis indicated that abundance of species foraging at mudflat level was positively associated with zone1 (winter) but negatively with zone3 (fall). Sandpipers were positively associated with zone4. Abundance distribution maps of conservation-related species, created by inverse distance-weighted interpolation modeling, also showed high abundance of most conservation-related species at zone4 and 1. The findings of our study suggest the importance of natural/semi-natural habitat, and the possible link between human activity and distribution patterns of migratory birds in Gochang getbol. While we need further investigation on direct response of migratory birds to human activity, areas with low human activity with more natural/semi-natural habitat, e.g., zone4 and zone1 may be crucial for the conservation of migratory birds.

Nitric oxide (NO) promotes angiogenesis via various mechanisms; however, the effective transmission of NO in ischemic diseases is unclear. Herein, we tested whether NO-releasing nanofibers modulate therapeutic angiogenesis in an animal hindlimb ischemia model. Male wild-type C57BL/6 mice with surgically-induced hindlimb ischemia were treated with NO-releasing 3-methylaminopropyltrimethoxysilane (MAP3)-derived or control (i.e., non-NO-releasing) nanofibers, by applying them to the wound for 20 min, three times every two days. The amount of NO from the nanofiber into tissues was assessed by NO fluorometric assay. The activity of cGMP-dependent protein kinase (PKG) was determined by western blot analysis. Perfusion ratios were measured 2, 4, and 14 days after inducing ischemia using laser doppler imaging. On day 4, Immunohistochemistry (IHC) with F4/80 and gelatin zymography were performed. IHC with CD31 was performed on day 14. To determine the angiogenic potential of NO-releasing nanofibers, aorta-ring explants were treated with MAP3 or control fiber for 20 min, and the sprout lengths were examined after 6 days. As per either LDPI (Laser doppler perfusion image) ratio or CD31 capillary density measurement, angiogenesis in the ischemic hindlimb was improved in the MAP3 nanofiber group; further, the total nitrate/nitrite concentration in the adduct muscle increased. The number of macrophage infiltrations and matrix metalloproteinase-9 (MMP-9) activity decreased. Vasodilator-stimulated phosphoprotein (VASP), one of the major substrates for PKG, increased phosphorylation in the MAP3 group. MAP3 nanofiber or NO donor SNAP (s-nitroso-n-acetyl penicillamine)-treated aortic explants showed enhanced sprouting in an ex vivo aortic ring assay, which was partially abrogated by KT5823, a potent inhibitor of PKG. These findings suggest that the novel NO-releasing nanofiber, MAP3 activates PKG and promotes therapeutic angiogenesis in response to hindlimb ischemia.

The alarming global rise in antibiotic resistance, driven by the widespread overuse of traditional antibiotics, has created an urgent demand for new antimicrobial solutions. This study presents zinc oxide (ZnO) nanorods as a potential nano-antibiotic agent. ZnO nanorods, with a 2:3 aspect ratio, were synthesized using an efficient one-step hydrothermal method at a low temperature of 100°C, reducing the synthesis time to just 5 hours. The synthesized ZnO nanorods' morphology, structure, and composition were characterized using scanning electron microscopy, x-ray diffraction, and energy dispersive x-ray spectroscopy. The potent antimicrobial activity of these nanorods against common bacterial strains such as Escherichia coli, Bacillus subtilis, and Vibrio parahaemolyticus was examined through optical density at 600 nm (OD600) measurements and inhibition zone analysis, demonstrating substantial inhibition of bacterial growth. In particular, at a concentration of 5 mg/mL, ZnO nanorods achieved a 96% reduction of B. subtilis bacteria in OD600 and an impressive 99.87% reduction in culturing assays within one day, showcasing bactericidal efficiency on par with tetracycline at 0.003 mg/mL. Furthermore, a predictive model of bacterial growth was developed and validated, providing insights into the time-dependent bactericidal efficiency of the synthesized nanorods. These results highlight the potential of ZnO-based composites as a promising solution to combat antibiotic resistance, paving the way for next-generation antimicrobial materials.

Beta cell failure is a critical feature of diabetes. It includes defects of insulin production, secretion, and altered numbers of hormone-producing cells. In previous work, we have shown that beta cell failure is mechanistically linked to loss of Foxo1 function. This loss of function likely results from increased Foxo1 protein degradation, due to hyperacetylation of Foxo1 from increased nutrient turnover. To understand the mechanisms of Foxo1-related beta cell failure, we performed genome-wide analyses of its target genes, and identified putative mediators of sub-phenotypes of cellular dysfunction. Chromatin immunoprecipitation analyses demonstrated a striking pattern of Foxo1 binding to the promoters of a cluster of aldo-ketoreductases on chromosome 13: Akr1c12, Akr1c13, Akr1c19. Of these, Akr1c19 has been reported as a marker of Pdx1-positive endodermal progenitor cells. Here we show that Akr1c19 expression is dramatically decreased in db/db islets. Thus, we investigated whether Akr1c19 is involved in beta cell function. We performed gain- and loss-of-function experiments in cultured beta cells and generated Akr1c19 knockout mice. We show that Foxo1 and HNF1a cooperatively regulate Akr1c19 expression. Nonetheless, functional characterization of Akr1c19 both using islets and knockout mice did not reveal abnormalities on glucose homeostasis. We conclude that reduced expression of Akr1c19 is not sufficient to affect islet function.

In this study, we present a novel approach to injection molding, focusing on the strength of weld lines in polyamide 6 (PA6) composite samples. By implementing a mold temperature significantly higher than the typical molding practice, which rarely exceeds 100°C, we assess the effects of advanced mold temperature management. The research introduces a newly engineered mold structure specifically designed for localized mold heating, distinguishing it as the 'novel cavity.' This innovative design is compared against traditional molding methods to highlight the improvements in weld line strength at elevated mold temperatures. To optimize the molding parameters, we apply an Artificial Neural Network (ANN) in conjunction with a Genetic Algorithm (GA). Our findings reveal that the optimal ultimate tensile strength (UTS) and elongation values are achieved with a filling time of 3.4 seconds, packing time of 0.8 seconds, melt temperature of 246°C, and a novel high mold temperature of 173°C. A specific sample demonstrated the best molding parameters at a filling time of 3.4 seconds, packing time of 0.4 seconds, melt temperature of 244°C, and mold temperature of 173°C, resulting in an elongation value of 582.6% and a UTS of 62.3 MPa. The most influential factor on the PA6 sample's UTS and elongation at the weld line was found to be the melt temperature, while the filling time had the least impact. SEM analysis of the fracture surfaces revealed ductile fractures with rough surfaces and grooves, indicative of the weld line areas' bonding quality. These insights pave the way for significant improvements in injection molding conditions, potentially revolutionizing the manufacturing process by enhancing the structural integrity of the weld lines in molded PA6 samples. [ABSTRACT FROM AUTHOR]

Background: In this study, the preliminary experience of endoscopic nipple-areolar-complex (NAC) sparing total mastectomy were analyzed and reported. Methods: The medical records of the patients who underwent Endoscopic NAC sparing total mastectomy from November 2019 to June 2022 in a single institute were collected and analyzed. The medical records of their clinicopathologic characteristics, perioperative parameters, postoperative complications, oncologic results were collected retrospectively. The learning curve was evaluated using univariable and multivariable analyses and analyzed using the CUSUM method. Results: A total of 56 surgeries from 53 patients were analyzed. The mean total operation time was 272.50±66.74 minutes while mean oncologic mastectomy was 155.61±43.21 minutes. The mean postoperative hospital days were 9.85±4.70 days. In CUSUM analysis, the cases needed to decrease operation time were 10th. The overall complication rate related to implant was 15.09%. There were one locoregional recurrence patient and one distant metastasis and mortality patient observed during median follow-up of 19.12±12.01 months. Conclusion: According to our preliminary study, endoscopic NAC sparing total mastectomy is a safe procedure, and the operation time can be reduced with accumulated surgical experience. [ABSTRACT FROM AUTHOR]

The ribosomal genes (rDNA genes) encode 47S rRNA which accounts for up to 80% of all cellular RNA. At any given time, no more than 50% of rDNA genes are actively transcribed, and the other half is silent by forming heterochromatin structures through DNA methylation. In cancer cells, upregulation of ribosome biogenesis has been recognized as a hallmark feature, thus, the reduced methylation of rDNA promoter has been thought to support conformational changes of chromatin accessibility and the subsequent increase in rDNA transcription. However, an increase in the heterochromatin state through rDNA hypermethylation can be a protective mechanism teetering on the brink of a threshold where cancer cells rarely successfully proliferate. Hence, clarifying hypo- or hypermethylation of rDNA will unravel its additional cellular functions, including organization of genome architecture and regulation of gene expression, in response to growth signaling, cellular stressors, and carcinogenesis. Using the bisulfite-based quantitative real-time methylation-specific PCR (qMSP) method after ensuring unbiased amplification and complete bisulfite conversion of the minuscule DNA amount of 1 ng, we established that the rDNA promoter was significantly hypermethylated in 107 breast, 65 lung, and 135 colon tumour tissue samples (46.81%, 51.02% and 96.60%, respectively) as compared with their corresponding adjacent normal samples (26.84%, 38.26% and 77.52%, respectively; p < 0.0001). An excessive DNA input of 1 μg resulted in double-stranded rDNA remaining unconverted even after bisulfite conversion, hence the dramatic drop in the single-stranded DNA that strictly required for bisulfite conversion, and leading to an underestimation of rDNA promoter methylation, in other words, a faulty hypomethylation status of the rDNA promoter. Our results are in line with the hypothesis that an increase in rDNA methylation is a natural pathway protecting rDNA repeats that are extremely sensitive to DNA damage in cancer cells. [ABSTRACT FROM AUTHOR]

The autonomous and active Long-Interspersed Element-1 (LINE-1, L1) and the non-autonomous Alu retrotransposon elements, contributing to 30% of the human genome, are the most abundant repeated sequences. With more than 90% of their sequences being methylated in normal cells, these elements undeniably contribute to the global DNA methylation level and constitute a major part of circulating-cell-free DNA (cfDNA). So far, the hypomethylation status of LINE-1 and Alu in cellular and extracellular DNA has long been considered a prevailing hallmark of ageing-related diseases and cancer. This study demonstrated that errors in LINE-1 and Alu methylation level measurements were caused by an excessive input quantity of genomic DNA used for bisulfite conversion. Using the minuscule DNA amount of 0.5 ng, much less than what has been used and recommended so far (500 ng-2 μg) or 1 μL of cfDNA extracted from 1 mL of blood, we revealed hypermethylation of LINE-1 and Alu in 407 tumour samples of primary breast, colon and lung cancers when compared with the corresponding pair-matched adjacent normal tissue samples (P < 0.05–0.001), and in cfDNA from 296 samples of lung cancers as compared with 477 samples from healthy controls (P < 0.0001). More importantly, LINE-1 hypermethylation in cfDNA is associated with healthy ageing. Our results have not only contributed to the standardized bisulfite-based protocols for DNA methylation assays, particularly in applications on repeated sequences but also provided another perspective for other repetitive sequences whose epigenetic properties may have crucial impacts on genome architecture and human health. [ABSTRACT FROM AUTHOR]

This paper investigates the performance of hybrid radio frequency/free space optical (RF/FSO) systems combined with non-orthogonal multiple access communications technology. We examine a scenario where the source and destination are separated by a large distance, with no direct link between them. The relay, denoted R, operates using the decode-and-forward (DF) protocol. Under the DF relaying scheme, the relay employs successive interference cancellation (SIC). In this setup, the FSO link from the source to the relay follows a Gamma-Gamma distribution, while the RF link from the relay to multiple users follow a Nakagami-m distribution. Based on this system model, we analyze the outage probability (OP). Our findings indicate a direct relationship between SIC and OP performance: the higher the SIC capability, the more effective the system. In addition, the system's performance is dependent on the parameters of the FSO channel. Finally, Monte Carlo simulations are presented to further validate our framework and findings. [ABSTRACT FROM AUTHOR]

It is important to establish the relationship between appropriate nutritional intake and improvements in activities of daily living (ADLs) in elderly hospitalized patients. This prospective observational study aimed to investigate diet order compliance (DOC) during 8 weeks of hospitalization and calculate the odds of improved functional independence measure (FIM) scores for high- and low-DOC groups using covariate-adjusted models in geriatric convalescent hospitals. The study subjects were elderly inpatients (>65 years old) with degenerative disease who consumed prescribed oral meals (Functional Oral Intake Scale (FOIS) = 6/7) and who did not receive physical/occupational therapy. The personalized diet order was prescript, and the DOC was calculated using dietitian-monitored daily intake data. The 73 patients were divided into a low-DOC group (< 84.0%, n = 35) and a high-DOC group (≥ 84.0%, n = 38) on the basis of the median DOC (84.0%, average for 8 weeks). Twenty (52.6%) high-DOC patients and nine (25.7%) low-DOC patients experienced motor-FIM improvements (P = 0.017). After 8 weeks, the change in motor-FIM in the high-DOC group (1.6±0.3) was greater than that in the low-DOC group (0.3±0.1; P = 0.001). According to the baseline and nutrition-intake-adjusted model of multiple logistic regression analysis, in the high-DOC group, the motor-FIM improvement OR was 5.102 (95% CI: 1.100–16.233, P = 0.036), and the total-FIM improvement OR was 5.273 (95% CI: 1.102–25.238, P = 0.037). High compliance with individualized nutritional prescriptions can increase FIM scores in clinical settings. Thus, comprehensive approaches to increase dietary compliance are needed for elderly long-term care patients. [ABSTRACT FROM AUTHOR]

Background: Hypertension (HTN) is linked to an enhanced risk of chronic kidney disease and cardiovascular disease. While sodium and potassium intake affect blood pressure (BP) control, the urine sodium-to-potassium (Na/K) ratio, which reflects dietary balance and renal regulation of these electrolytes, could be associated with BP. This study aimed to evaluate the independent association between urine Na/K and uncontrolled HTN. Methods: Data were collected from the Korea National Health and Nutrition Examination Survey from 2016 to 2021. A total of 5,770 participants diagnosed with HTN were enrolled in this study. Uncontrolled HTN was characterized by a systolic blood pressure (SBP) ≥ 140 mmHg or diastolic blood pressure (DBP) ≥ 90 mmHg. Logistic regression analysis was used to assess the relationship between urine Na/K and the risk of uncontrolled HTN. Results: The urine Na/K was positively correlated with both SBP and mean arterial pressure. Higher urine Na/K was significantly associated with an increased risk of uncontrolled HTN using both continuous (odds ratio [95% confidence interval] 1.13 [1.09–1.16], P <0.01]) and across quartile values (with Q1 as a reference; Q2: 1.26 [1.06–1.49], P = 0.01; Q3: 1.50 [1.27–1.78], P <0.01; Q4: 1.85 [1.55–2.17], P < 0.01). The subgroup analysis also showed that higher urine Na/K were significantly related to the risk of uncontrolled HTN in the presence of proteinuria or CKD. Conclusion: Urine Na/K ratio is independently associated with uncontrolled HTN in the general population and in patients with CKD. Our findings suggest that monitoring the urine Na/K could serve as an effective tool for identifying subjects at risk of uncontrolled HTN. [ABSTRACT FROM AUTHOR]

The alarming global rise in antibiotic resistance, driven by the widespread overuse of traditional antibiotics, has created an urgent demand for new antimicrobial solutions. This study presents zinc oxide (ZnO) nanorods as a potential nano-antibiotic agent. ZnO nanorods, with a 2:3 aspect ratio, were synthesized using an efficient one-step hydrothermal method at a low temperature of 100°C, reducing the synthesis time to just 5 hours. The synthesized ZnO nanorods' morphology, structure, and composition were characterized using scanning electron microscopy, x-ray diffraction, and energy dispersive x-ray spectroscopy. The potent antimicrobial activity of these nanorods against common bacterial strains such as Escherichia coli, Bacillus subtilis, and Vibrio parahaemolyticus was examined through optical density at 600 nm (OD600) measurements and inhibition zone analysis, demonstrating substantial inhibition of bacterial growth. In particular, at a concentration of 5 mg/mL, ZnO nanorods achieved a 96% reduction of B. subtilis bacteria in OD600 and an impressive 99.87% reduction in culturing assays within one day, showcasing bactericidal efficiency on par with tetracycline at 0.003 mg/mL. Furthermore, a predictive model of bacterial growth was developed and validated, providing insights into the time-dependent bactericidal efficiency of the synthesized nanorods. These results highlight the potential of ZnO-based composites as a promising solution to combat antibiotic resistance, paving the way for next-generation antimicrobial materials. [ABSTRACT FROM AUTHOR]

BackgroundSARS-CoV-2 Delta variant caused a large number of COVID-19 cases in many countries, including Vietnam. Understanding mortality risk factors is crucial for the clinical management of severe COVID-19.MethodsWe conducted a retrospective study at an intensive care center in Ho Chi Minh City that urgently built by Bach Mai Hospital during the COVID-19 outbreak in Vietnam, when the Delta variant predominated. Participants were laboratory-confirmed patients with SARS-CoV-2 infection, admitted in August 2021. Data on patients' demographic and clinical characteristics, radiographic and laboratory findings, treatment, and clinical time course were compared between survivors and non-survivors. Risk factors to mortality were assessed using logistic regression.ResultsAmong 504 eligible COVID-19 patients, case fatality was 52.2%. Unvaccinated patients accounted for 61.2% of non-survivors and 43.6% of survivors (p < 0.001). The time from onset to hospital admission was 8 days in non-survivors and 7 days in survivors (p = 0.004). Among non-survivors, 90.2% developed acute respiratory distress syndrome (ARDS). Oxygen therapy was administered for all patients, but antiviral agent was given to 51.7% of non-survivors. 54.2% of non-survivors tested positive for the bacterial infection using blood culture. The risk factors for mortality were diabetes mellitus, respiration rate, oxygen saturation, vaccination status, time from onset to admission, and older age.ConclusionsCritical patients with COVID-19 owing to the Delta variant in Vietnam had delayed hospital admission, leading to ARDS and death. Early availability of vaccines and preventing bacterial infections are crucial for reducing mortality of COVID-19, especially in low- and middle-income countries.

Background Growing evidence suggests an association between the vitamin D levels and respiratory outcomes of preterm infants. The objective of this systematic review and meta-analysis was to explore whether premature neonates with a vitamin D deficiency have an increased risk of respiratory distress syndrome (RDS). Methods We searched PubMed, EMBASE, and the Cochrane Library up through July 20, 2021. The search terms were ‘premature infant’, ‘vitamin D’, and ‘respiratory distress syndrome’. We retrieved randomized controlled trials and cohort and case-control studies. For statistical analysis, we employed the random-effects model in Comprehensive Meta-Analysis Software ver. 3.3. We employed the Newcastle-Ottawa Scales for quality assessment of the included studies. Results A total of 121 potentially relevant studies were found, of which 15 (12 cohort studies and 3 case-control studies) met the inclusion criteria; the studies included 2,051 preterm infants. We found significant associations between RDS development in such infants and vitamin D deficiency within 24 h of birth based on various criteria, thus vitamin D levels < 30 ng/mL (OR 3.478; 95% CI 1.817–6.659; p < 0.001), < 20 ng/mL (OR 4.549; 95% CI 3.007–6.881; p < 0.001), < 15 ng/mL (OR 17.267; 95% CI 1.084–275.112; p = 0.044), and < 10 ng/ml (OR 1.732; 95% CI 1.031–2.910; p = 0.038), and an even lower level of vitamin D (SMD = –0.656; 95% CI –1.029 to –0.283; p = 0.001). Conclusion Although the vitamin D deficiency definitions varied and different methods were used to measure vitamin D levels, vitamin D deficiency or lower levels of vitamin D within 24 h of birth were always associated with RDS development. Monitoring of neonatal vitamin D levels or the maintenance of adequate levels may reduce the risk of RDS.

Purpose To assess experience with and perceptions of clinical application of artificial intelligence (AI) to chest radiographs among doctors in a single hospital. Materials and methods A hospital-wide online survey of the use of commercially available AI-based lesion detection software for chest radiographs was conducted with all clinicians and radiologists at our hospital in this prospective study. In our hospital, version 2 of the abovementioned software was utilized from March 2020 to February 2021 and could detect three types of lesions. Version 3 was utilized for chest radiographs by detecting nine types of lesions from March 2021. The participants of this survey answered questions on their own experience using AI-based software in daily practice. The questionnaires were composed of single choice, multiple choices, and scale bar questions. Answers were analyzed according to the clinicians and radiologists using paired t-test and the Wilcoxon rank-sum test. Results One hundred twenty-three doctors answered the survey, and 74% completed all questions. The proportion of individuals who utilized AI was higher among radiologists than clinicians (82.5% vs. 45.9%, p = 0.008). AI was perceived as being the most useful in the emergency room, and pneumothorax was considered the most valuable finding. Approximately 21% of clinicians and 16% of radiologists changed their own reading results after referring to AI, and trust levels for AI were 64.9% and 66.5%, respectively. Participants thought AI helped reduce reading times and reading requests. They answered that AI helped increase diagnostic accuracy and were more positive about AI after actual usage. Conclusion Actual adaptation of AI for daily chest radiographs received overall positive feedback from clinicians and radiologists in this hospital-wide survey. Participating doctors preferred to use AI and regarded it more favorably after actual working with the AI-based software in daily clinical practice.

Sexually transmitted diseases are major causes of infertility, ectopic pregnancy, and premature birth. Here, we developed a new multiplex real-time polymerase chain reaction (PCR) assay for the simultaneous detection of nine major sexually transmitted infections (STIs) found in Vietnamese women, including Chlamydia trachomatis, Neisseria gonorrhoeae, Gardnerella vaginalis, Trichomonas vaginalis, Candida albicans, Mycoplasma hominis, Mycoplasma genitalium, and human alphaherpesviruses 1 and 2. A panel containing three tubes × three pathogens/tube was predesigned based on double-quenched TaqMan probes to increase detection sensitivity. There was no cross-reactivity among the nine STIs and other non-targeted microorganisms. Depending on each pathogen, the agreement with commercial kits, sensitivity, specificity, repeatability and reproducibility coefficient of variation (CV), and limit of detection of the developed real-time PCR assay were 99.0%–100%, 92.9%–100%, 100%,

The measurement of skeletal muscle mass is essential for the diagnosis of sarcopenia. Muscle ultrasonography has emerged as a useful tool for evaluating sarcopenia because it can be used to assess muscle quality and quantity. This study investigated whether muscle ultrasonography is effective for estimating appendicular skeletal muscle mass (ASM) and screening for sarcopenia. This study prospectively enrolled 212 healthy volunteers aged 40-80 years. ASM was measured using the bioelectrical impedance analysis. Muscle thickness (MT) and echo-intensity (EI) were measured in four muscles (biceps brachii, BB; triceps brachii, TB; rectus femoris, RF; biceps femoris, BF) on the dominant hand. A hold-out cross-validation method was used to develop and validate the ASM prediction equation. In the model development group, the ASM prediction equations were deduced as follows: estimated ASM for men (kg) = 0.167 × weight (kg) + 0.228 × height (cm) + 0.143 × MT of BF (mm)- 0.822 × EI to MT ratio of BB- 28.187 (R2 = 0.830) and estimated ASM for women (kg) = 0.115 × weight + 0.215 × height (cm) + 0.139 × MT of RF-0.638 × EI to MT ratio of BB- 23.502 (R2 = 0.859). In the cross-validation group, the estimated ASM did not significantly differ from the measured ASM in both men (p = 0.775; intraclass correlation coefficient [ICC] = 0.948) and women (p = 0.516; ICC = 0.973). In addition, multiple logistic regression analysis revealed that the ratios of EI to MT in the BF and RF muscles in men and MT in the BB muscle in women could be valuable parameters for sarcopenia screening. Therefore, our study suggests that muscle ultrasound could be an effective tool for estimating ASM and screening sarcopenia.

ObjectivesThe objectives of this study were 1) to investigate the prevalence and co-existence of frailty and malnutrition and 2) to identify factors related to frailty (including malnutrition) according to the level of frailty.MethodsData collection was conducted from July 11, 2021, to January 23, 2022, in 558 older adults residing in 16 long-term care facilities (LTCFs) in Korea. The FRAIL-NH and Mini-Nutritional Assessment short form were used to measure frailty and nutrition, respectively. The data analysis included descriptive statistics and a multivariate logistic regression.ResultsThe mean age of the participants was 83.68 (± 7.39) years. Among 558 participants, 37 (6.6%), 274 (49.1%), and 247 (44.3%) were robust, prefrail, and frail, respectively. At the same time, 75.8% were categorized as having malnutrition status (malnourished: 18.1%; risk of malnutrition: 57.7%), and 40.9% had co-existing malnutrition and frailty. In the multivariate analysis, malnutrition was identified as the major frailty-related factor. Compared with a normal nutritional status, the incidence of frailty in the malnutrition group was 10.35 times (95% CI: 3.78-28.36) higher than the incidence of robustness and 4.80 times (95% CI: 2.69-8.59) higher than the incidence of prefrail.ConclusionThe prevalence of frailty and malnutrition, and their co-existence, among older adults residing in LTCFs was high. Malnutrition is a major factor that increases the incidence of frailty. Therefore, active interventions are needed to improve the nutritional status of this population.

BackgroundTaxation on sugar-sweetened beverages (SSBs) has been adopted in more than 40 countries but remained under discussion in Vietnam. This study aimed to estimate the health impacts of different SSBs tax plans currently under discussion to provide an evidence base to inform decision-making about a SSBs tax policy in Vietnam.Method & findingsFive tax scenarios were modelled, representing three levels of price increase: 5%, 11% and 19-20%. Scenarios of the highest price increase were assessed across three different tax designs: ad valorem, volume-based specific tax & sugar-based specific tax. We modelled SSBs consumption in each tax scenario; how this reduction in consumption translates to a reduction in total energy intake and how this relationship in turn translates to an average change in body weight and obesity status among adults by applying the calorie-to weight conversion factor. Changes in type 2 diabetes burden were then calculated based on the change in average BMI of the modelled cohort. A Monte Carlo simulation approach was applied on the conversion factor of weight change and diabetes risk reduction for the sensitivity analysis. We found that the taxation that involved a 5% price increase gave relatively small impacts while increasing SSBs' price up to 20% appeared to impact substantially on overweight and obesity rates (reduction of 12.7% and 12.4% respectively) saving 27 million USD for direct medical cost. The greatest reduction was observed for overweight and obesity class I. The decline in overweight and obesity rates was slightly higher for women than men.ConclusionThis study supports the SSB tax policy in pursuit of public health benefits, especially where the tax increase involves around a 20% price increase. The health benefit and revenue gains were evident across all three tax designs with the specific tax based on sugar density achieving greatest effects.