Application of Directed Evolution and Machine Learning to Enhance the Diastereoselectivity of Ketoreductase for Dihydrotetrabenazine Synthesis.

Biocatalysis is an effective approach for producing chiral drug intermediates that are often difficult to synthesize using traditional chemical methods. A time-efficient strategy is required to accelerate the directed evolution process to achieve the desired enzyme function. In this research, we evaluated machine learning-assisted directed evolution as a potential approach for enzyme engineering, using a moderately diastereoselective ketoreductase library as a model system. Machine learning-assisted directed evolution and traditional directed evolution methods were compared for reducing (±)-tetrabenazine to dihydrotetrabenazine via kinetic resolution facilitated by BsSDR10, a short-chain dehydrogenase/reductase from Bacillus subtilis. Both methods successfully identified variants with significantly improved diastereoselectivity for each isomer of dihydrotetrabenazine. Furthermore, the preparation of (2S,3S,11bS)-dihydrotetrabenazine has been successfully scaled up, with an isolated yield of 40.7% and a diastereoselectivity of 91.3%.

Comparisons of Various Anthropometric Indices for Incident Hypertension in Military Young Adults: The CHIEF Cohort Study.

AIM: The study aimed to compare the predictive capabilities of the traditional anthropometric indices with the novel anthropometric indices for incident hypertension. BACKGROUND: Some novel anthropometric indices, e.g., the Body Roundness Index (BRI) and A Body Shape Index (ABSI) have been associated with prevalent hypertension. There are a few cohort studies that have examined the association of the novel anthropometric indices with newonset hypertension in young adults. METHODS: This study included 2,448 military male and female young adults, aged 18-39 years, free of hypertension at baseline in Taiwan; they were followed for incidence of hypertension from 2014 till the end of 2020. Blood Pressure (BP) in mmHg was measured twice and averaged to verify hypertension, which was defined as systolic BP ≥130 and/or diastolic BP ≥80 or on antihypertensive medication therapy in each annual health examination. Anthropometric indices included the Body Mass Index (BMI) defined as the weight (kg)/height squared (m2), Waist Girth (WC) in cm, the Waist-to-height Ratio (WHtR), the BRI defined as 364.2 - 365.5 × {1 - [(WC/2π)/(0.5 × height)]2}0.5, as well as ABSI defined as WC/(BMI2/3 × height1/2). Multiple Cox regression analysis and Area Under the Curve (AUC) of the Receiver of Operating Characteristics (ROC) were utilized with adjustments for the baseline potential covariates to determine the association and compare the performance of various indices for incident hypertension. RESULTS: During a median follow-up period of 6.0 years, 920 new-onset hypertension cases (37.6%) developed. Higher BMI, WC, BRI (per each 1-unit increase) and WHtR (per each 0.1- unit increase) were associated with a greater risk of new-onset hypertension [Hazard Ratios (HRs) and 95% confidence intervals: 1.060 (1.035-1.085), 1.021 (1.011-1.030), and 1.178 (1.077-1.288), respectively], whereas there was no association between ABSI and new-onset hypertension. For the ROC, WC was observed with the greatest AUC for incident new-onset hypertension [0.661 (0.638-0.683)], followed by BMI [0.650 (0.628-0.673)], while the ABSI was found with the lowest AUC [0.544 (0.521-0.568)]. CONCLUSION: Most of the anthropometric indices were associated with a higher risk of new-onset hypertension among young adults, except for ABSI. In addition, this study has suggested the traditional indices, such as WC and BMI, to be superior to the latest ones, e.g., BRI and ABSI, for the prediction of new-onset hypertension.

Asymmetric Synthesis of ß-Hydroxyphosphonates via a Chemoenzymatic Cascade.

Chiral ß-hydroxyphosphonates are essential building blocks for organophosphorus compounds. However, the asymmetric synthesis of these units remains a significant challenge. Herein, we describe a one-pot chemoenzymatic cascade process to access chiral ß-hydroxyphosphonates, which combines photo-oxidative chemical reactions and bioreductions. The incorporation of photooxidation in the chemical reaction resulted in up to 92% yield and >99% enantiomeric excess (ee) of ß-hydroxyphosphonates in the cascade. In addition, the scale-up of diethyl (S)-(2-hydroxy-2-phenylethyl)phosphonate demonstrates the potential application of this strategy.

Does the guideline-based physical activity level for cardiovascular health also benefit periodontal health?

Abstract Background/purpose: It is unclear about whether the guideline-based physical activity (PA) level for cardiovascular health also benefits periodontal health. Therefore, this study aimed to clarify the association between guideline-based PA levels and periodontitis in young adults. Materials and methods: This was a cross-sectional study which included 334 military participants, aged 18-44 years in Taiwan. The PA level was assessed by total running time per week (wk) in the past 6 months from a self-reported questionnaire, and was classified as moderate intensity PA <150, 150-299, and ≥300 min/wk according to the American guideline for cardiovascular health. Periodontitis was defined based on the 2017 World Workshop. Multiple logistic regression analysis was used to determine the association with adjustment for age, sex, educational level, tobacco smoking, alcohol intake, body mass index, waist circumference, blood pressure and teeth brushing frequency. Results: As compared to the PA: 150-299 min/wk, the PA <150 min/wk was associated with a greater possibility of periodontitis (odds ratio (OR): 2.45 (95% confidence interval (CI): 1.24-4.82), whereas the PA ≥300 min/wk was not associated with periodontitis (OR: 0.98 (95% CI: 0.13-1.09)) in young adults. Conclusion: The guideline-based moderate intensity PA level ≥150 min/wk is found to have superior periodontal health in young adults. However, there is no additional benefit in more exercise time to obtain better periodontal health when the suggested PA level has been achieved.

Obesity Phenotypes and Dental Calculus in Young Adults: CHIEF Oral Health Study.

AIM: The study aimed to examine the association of obesity phenotypes with dental calculus. BACKGROUND: Obesity has been recognized as a risk factor for kidney and gallbladder stones formation and periodontitis. OBJECTIVE: We have investigated the association between obesity, metabolic risk factors, and dental calculus, which is a sequela following periodontitis. METHODS: This study included 5,281 military members, aged 19-45 years, without antihypertensive medications in Taiwan. Obesity was defined as body mass index ≥27.5 kg/m2, and metabolic syndrome (MetS) was defined according to the modified ATP III criteria. Supragingival calculus in any teeth, except for impacted teeth and the third molar, was the outcome of interest. Multiple linear regression analysis with adjustments for age, sex, toxic substance use, brushing teeth frequency, and blood leukocyte counts, was used to determine the association of obesity with dental calculus numbers. Multiple logistic regression analysis was used to assess the association between obesity with or without MetS and the presence of any dental calculus. RESULTS: BMI was positively correlated to dental calculus numbers [ß and confidence intervals (CI) = 0.023 (0.014, 0.032)]. Compared to the obesity(-)/MetS(-) group, there were dosedependent associations for the obesity(-)/MetS(+), obesity(+)/MetS(-), and obesity(+)/MetS(+) groups with the presence of any dental calculus [odds ratios (ORs): 1.08 (0.76, 1.53), 1.31 (1.08, 1.58), and 1.51 (1.20, 1.90), respectively]. Of the metabolic risk factors, abdominal obesity and hypertension were independently associated with dental calculus [ORs: 1.33 (1.13, 1.55) and 1.30 (1.11, 1.52), respectively]. CONCLUSION: This study suggests general obesity as an independent risk factor for dental calculus formation, and MetS, particularly the components of abdominal obesity, and hypertension may also increase the prevalence of dental calculus. Diet control and regular exercise might be preventive measures for the development of both obesity and dental calculus.

Quantitative multiphoton imaging of cell metabolism, stromal fibers, and keratinization enables label-free discrimination of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) features atypical clinical manifestations and a low 5-year survival rate (< 5% in many developing countries where most of the disease occurs). Precise ESCC detection and grading toward timely and effective intervention are therefore crucial. In this study, we propose a multidimensional, slicing-free, and label-free histopathological evaluation method based on multispectral multiphoton fluorescence lifetime imaging microscopy (MM-FLIM) for precise ESCC identification. To assess the feasibility of this method, comparative imaging on fresh human biopsy specimens of different ESCC grades is performed. By constructing fluorescence spectrum- and lifetime-coded images, ESCC-induced morphological variations are unveiled. Further quantification of cell metabolism and stromal fibers reveals potential indicators for ESCC detection and grading. The specific identification of keratin pearls provides additional support for the early detection of ESCC. These findings demonstrate the viability of using MM-FLIM and the series of derived indicators for histopathological evaluation of ESCC. As there is an increasing interest in developing multiphoton endoscopes and multiphoton FLIM systems for clinical use, the proposed method would probably allow noninvasive, label-free, and multidimensional histological detection and grading of ESCC in the future.

Epidemiological characteristics, clinical manifestations, and mental health status of human mpox cases: A multicenter cross-sectional study in China.

Human mpox is occurring worldwide, however, evidence from the Asian Pacific Region is limited. In this multicenter cross-sectional study, information of confirmed mpox cases diagnosed between June 1 and July 31, 2023 in China. Information included demographic and epidemiological characteristics, and clinical manifestations, laboratory results, and mental health status of mpox cases. A total of 115 confirmed mpox cases were enrolled. All cases were men. A total of 102 (90.3%) identified as homosexual. The median age was 31.0 years (interquartile range 27.0-36.5). A total of 65 (56.5%) were HIV-positive, of whom 92.3% were receiving antiretroviral therapy (ART). A total of 19/39 (40.4%) had a CD4 cell count <500 cells/µL. Systemic features such as fever (73.0%), lymphadenopathies (49.6%), and myalgia (28.7%) were commonly observed. Skin lesions were present in all participants: 49.6% in the genital area and 27.0% in the perianal area. Vesicular rash (78.3%) and papular rash (44.3%) were the most common lesion morphologies. People living with HIV were more likely to have anxiety than those living without HIV. The majority of mpox cases had primary genital lesions and sexual activities before diagnosis, which supports the likelihood of sexual contact transmission. Guidelines on hospitalization and isolation protocols for mpox patients necessitate further confirmation.

Programmable Synthesis of High-Entropy Nanoalloys for Efficient Ethanol Oxidation Reaction.

Controllable synthesis of nanoscale high-entropy alloys (HEAs) with specific morphologies and tunable compositions is crucial for exploring advanced catalysts. The present strategies either have great difficulties to tailor the morphology of nanoscale HEAs or suffer from narrow elemental distributions and insufficient generality. To overcome the limitations of these strategies, here we report a robust template-directed synthesis to programmatically fabricate nanoscale HEAs with controllable compositions and structures via independently controlling the morphology and composition of HEA. As a proof of concept, 12 kinds of nanoscale HEAs with controllable morphologies of zero-dimension (0D) nanoparticles, 1D nanowires, 2D ultrathin nanorings (UNRs), 3D nanodendrites, and vast elemental compositions combining five or more of Pd/Pt/Ag/Cu/Fe/Co/Ni/Pb/Bi/Sn/Sb/Ge are synthesized. Moreover, the as-prepared HEA-PdPtCuPbBiUNRs/C demonstrates the state-of-the-art electrocatalytic performance for the ethanol oxidation reaction, with 25.6- and 16.3-fold improvements in mass activity, relative to commercial Pd/C and Pt/C catalysts, respectively, as well as greatly enhanced durability. This work provides a myriad of nanoscale HEAs and a general synthetic strategy, which are expected to have broad impacts for the fields of catalysis, sensing, biomedicine, and even beyond.

Rapid and label-free histological imaging of unprocessed surgical tissues via dark-field reflectance ultraviolet microscopy.

Routine examination for intraoperative histopathologic assessment is lengthy and laborious. Here, we present the dark-field reflectance ultraviolet microscopy (DRUM) that enables label-free imaging of unprocessed and thick tissues with subcellular resolution and a high signal-to-background ratio. To the best of our knowledge, DRUM provides image results for pathological assessment with the shortest turnaround time (2-3 min in total from sample preparation to tissue imaging). We also proposed a virtual staining process to convert DRUM images into pseudo-colorized images and enhance the image familiarity of pathologists. By imaging various tissues, we found DRUM can resolve cell nuclei and some extranuclear features, which are comparable to standard H&E images. Furthermore, the essential diagnostic features of intraoperatively excised tumor tissues also can be revealed by DRUM, demonstrating its potential as an additional aid for intraoperative histopathology.

Ketoreductase Catalyzed (Dynamic) Kinetic Resolution for Biomanufacturing of Chiral Chemicals.

Biocatalyzed asymmetric reduction of ketones is an environmentally friendly approach and one of the most cost-effective routes for producing chiral alcohols. In comparison with the well-studied reduction of prochiral ketones to generate chiral alcohols with one chiral center, resolution of racemates by ketoreductases (KREDs) to produce chiral compounds with at least two chiral centers is also an important strategy in asymmetric synthesis. The development of protein engineering and the combination with chemo-catalysts further enhanced the application of KREDs in the efficient production of chiral alcohols with high stereoselectivity. This review discusses the advances in the research area of KRED catalyzed asymmetric synthesis for biomanufacturing of chiral chemicals with at least two chiral centers through the kinetic resolution (KR) approach and the dynamic kinetic resolution (DKR) approach.

The Sandwich Technique with Body Flossing Wire to Revascularize Left Subclavian Artery in Thoracic Endovascular Aortic Repair.

BACKGROUND: To evaluate the safety and efficacy of the sandwich technique with the body flossing wire to revascularize the left subclavian artery (LSA) in thoracic endovascular aortic repair (TEVAR). METHODS: From April 2014 to April 2015, 20 consecutive patients with a variety of thoracic aortic pathologies who underwent LSA revascularization with the sandwich technique and body flossing wire during TEVAR were included. Outcomes including technical success, endoleaks, perioperative mortality and morbidity, and graft patency were analyzed. RESULTS: There were 13 patients (65.0%) who presented with aortic dissection, 3 (15.0%) with thoracic aortic aneurysm, 3 (15.0%) with penetrating aortic ulcer, and 1 (5.0%) with traumatic aortic disruption. Technical success rate was 100%. But one type Ia endoleak (5.0%) was noted and treated with extended endografting. One sandwiched gutter leak (5.0%) resolved spontaneously at 6 months. One sandwiched graft (5.0%) was occluded but not treated because no related complications were found. At a mean follow-up of 9.7 months (range 4-17), all the patients with aortic aneurysm, penetrating aortic ulcer, or traumatic aortic disruption had complete thrombosis of the aortic pathologies. Patients with aortic dissection had thrombosed false lumen down to the distal aortic endograft edge (12/13, 92.3%) or the celiac artery level (8/13, 61.5%). There was no neurological deficit but 2 non-procedure-related late deaths during the study period. CONCLUSIONS: Our early experience showed that the sandwich technique with the body flossing wire to maintain blood flow to the LSA is a safe and effective method. This technique can be applied in various clinical situations with a high technique success rate. However, outcomes are preliminary and larger studies are required.

Isolated intestinal transplantation for megacystis microcolon intestinal hypoperistalsis syndrome: case report.

MMIHS is a rare congenital disease. It is characterized by distended urinary bladder, small colon and intestinal hypoperistalsis, or aperistalsis with normal morphology. There is no specific treatment for MMIHS, and most patients have to be maintained by TPN, which frequently causes TPN-related liver failure, loss of venous access, or catheter-related sepsis. The prognosis of patients with MMIHS is poor, and most patients die early. Multivisceral transplantation including stomach, duodenum, intestine, and liver has been used for the treatment of patients with MMIHS because these patients often have liver failure. We report an eight-yr-old patient with MMIHS who was treated with isolated intestinal transplantation. She had completely oral intake during the four yr of follow-up. The experience in this case suggests isolated intestinal transplantation may be indicated in selected cases with MMIHS.