ST-ITEF: Spatio-Temporal Intraoperative Task Estimating Framework to recognize surgical phase and predict instrument path based on multi-object tracking in keratoplasty.

Computer-assisted cognition guidance for surgical robotics by computer vision is a potential future outcome, which could facilitate the surgery for both operation accuracy and autonomy level. In this paper, multiple-object segmentation and feature extraction from this segmentation are combined to determine and predict surgical manipulation. A novel three-stage Spatio-Temporal Intraoperative Task Estimating Framework is proposed, with a quantitative expression derived from ophthalmologists' visual information process and also with the multi-object tracking of surgical instruments and human corneas involved in keratoplasty. In the estimation of intraoperative workflow, quantifying the operation parameters is still an open challenge. This problem is tackled by extracting key geometric properties from multi-object segmentation and calculating the relative position among instruments and corneas. A decision framework is further proposed, based on prior geometric properties, to recognize the current surgical phase and predict the instrument path for each phase. Our framework is tested and evaluated by real human keratoplasty videos. The optimized DeepLabV3 with image filtration won the competitive class-IoU in the segmentation task and the mean phase jaccard reached 55.58 % for the phase recognition. Both the qualitative and quantitative results indicate that our framework can achieve accurate segmentation and surgical phase recognition under complex disturbance. The Intraoperative Task Estimating Framework would be highly potential to guide surgical robots in clinical practice.

Docosahexaenoic acid alleviates LPS-induced cytotoxicity in HL-1 cardiac cells via improving stress-induced mitochondrial fragmentation.

Sepsis-induced cardiac injury is associated with oxidative stress and mitochondrial dysfunction. Docosahexaenoic acid (DHA), an essential omega-3 fatty acid, protects the injured myocardium by modulating mitochondrial dysfunction. We aimed to confirm whether the cardioprotective effect of DHA is mediated via the alleviation of mitochondrial fragmentation in lipopolysaccharide (LPS)-induced cardiomyopathy in vitro. We found that DHA improved cell viability and alleviated cardiac cell apoptosis by reducing lactate dehydrogenase (LDH) release, expression levels of Cleaved caspase-3, and Caspase 3 activity. DHA attenuated oxidative stress as evidenced by decreased ROS production and increased superoxide dismutase activity. In addition, DHA ameliorated mitochondrial dysfunction by modulating mitochondrial respiratory chain injury and mitochondrial fragmentation, especially decreasing the mitochondrial fission-related protein p-Drp1(ser 616) but no effects on Drp1, p-Drp1(ser 637), and mitochondrial fusion-related protein. Our data suggest that DHA conferred cardioprotection by alleviating oxidative stress-induced apoptosis, which may be associated with alleviation of stress-induced mitochondrial fragmentation.

GSDMD-mediated pyroptosis restrains intracellular Chlamydia trachomatis growth in macrophages.

Pyroptosis, a type of programmed necrosis associated with inflammatory, is a host defense mechanism against microbial infections. Although Chlamydia has been shown to induce pyroptosis, whether pyroptosis directly impacts the growth of Chlamydia has not been demonstrated. In this study, we found that C. trachomatis L2 infection of the mouse macrophage RAW 264.7 cells induced pyroptosis by monitoring the ultrastructural changes under transmission electron microscopy and the release of LDH and IL-1ß. More importantly, this C. trachomatis-triggered pyroptosis with activation of caspase-1 and caspase-11 was also accompanied by gasdermin D (GSDMD) activation. Suppression of these two inflammatory caspases inhibited GSDMD activation. Interestingly, the C. trachomatis-triggered pyroptosis significantly inhibited the intracellular growth of C. trachomatis since inactivation of either GSDMD or caspase-1/11 significantly rescued infectious C. trachomatis yields, which suggests pyroptosis response can be utilized as an intrinsic mechanism to restrict C. trachomatis intracellular infection in addition to the well- documented extrinsic mechanisms by recruiting and enhancing inflammatory responses. This study may reveal novel targets for attenuating C. trachomatis infectivity and/or pathogenicity.

The Association Between Hyperuricemia and Obesity Metabolic Phenotypes in Chinese General Population: A Retrospective Analysis.

Purpose: Serum uric acid (UA) not only affects the development of obesity but also alters the metabolic status in obese subjects; thus we investigated the relationship between serum UA and the overweight/obese metabolic phenotypes. Methods: The demographic, biochemical, and hematological data were collected for 12,876 patients undergoing routine physical examination, and 6,912 participants were enrolled in our study. Participants were classified into four obesity metabolic phenotypes according to their BMI and the presence of metabolic syndrome: metabolically healthy overweight/obese (MHOO), metabolically healthy and normal weighted (MHNW), metabolically abnormal and overweight/obese (MAOO), and metabolically abnormal but normal weighted (MANW). Univariate and multivariate logistic regression analysis, stratified analysis, and also interaction analysis were conducted to analyze the relationship between serum UA and obesity metabolic phenotypes. Results: Multivariable logistic regression analysis showed that hyperuricemia was positively associated with MHOO, MANW, and MAOO phenotypes relative to MHNW. After adjusting for the confounding factors, the odds ratios (OR) for individuals with hyperuricemia to be MHOO, MANW, and MAOO phenotypes were 1.86 (1.42-2.45), 2.30 (1.44-3.66), and 3.15 (2.34-4.24), respectively. The ORs for having MHOO, MANW, and MAOO increased 6% [OR: 1.06 (1.05-1.07), P < 0.0001], 5% [OR: 1.05 (1.03-1.07), P < 0.0001], and 11% [OR: 1.11 (1.10-1.13), P < 0.0001] for each 10 unit (µmol/L) of increase in serum UA level. Stratification analysis as well as an interaction test showed that sex and age did not interfere with the association of hyperuricemia with each metabolic phenotype. In terms of the components of the metabolic syndrome, after adjusting for other confounding factors including all of the metabolic indicators except itself, hyperuricemia was positively associated with increased BMI [OR: 1.66 (1.32-2.09), P < 0.0001], hypertriglyceridemia [OR: 1.56 (1.21-2.02), P = 0.0006], and hypertension [OR: 1.22 (1.03-1.46), P = 0.0233], while it had no significant association with hyperglycemia and low HDL-C (all P > 0.05). Conclusion: In our study, we discovered that hyperuricemia was positively associated with MHOO, MANW, and MAOO phenotypes, and this relationship was independent of sex and age.

REPORT-Mild encephalitis/encephalopathy with a reversible splenial lesion associated with mumps infection: A case report.

We describe for the first time an child who demonstrated Mild encephalitis/encephalopathy with a reversible splenial lesion (MERS) after mumps infection in China. In this report, a 12-year-old boy came to Children's Hospital Affiliated to Zhengzhou University due to fever, swelling and pain under the earlobe for 4 days, and headache and vomiting for half of a day. Laboratory examinations showed a blood sodium level of 125mmol/L, both the Immunoglobulin M and Polymerase Chain Reaction results for the serum mumps virus were positive. Brain Magnetic Resonance Imaging (MRI) showed slight hypointense on T1 weighted images, hyperintense on T2-weighted images, fluid attenuated inversion recovery, diffusion-weighted images in the splenium of the corpus callosum indicative of MERS. On the 8th day, the patient no longer had swelling and pain around the parotid salivary glands, the sodium levels returned to normal. Onset of 14th d, follow-up brain MRI did not reveal any abnormalities. The case given to us indicates that MERS should be considered when patients after mumps infection presents with neurological symptoms and MRI should be performed to evaluate the splenium of the corpus callosum.

Genome-wide high-resolution mapping of DNA methylation reveals epigenetic variation in the offspring of sexual and asexual propagation in Robinia pseudoacacia.

KEY MESSAGE: We detected the genome-wide pattern of DNA methylation and its association with gene expression in sexual and asexual progenies of mature Robinia pseudoacacia trees. DNA methylation plays an important role in plant reproduction and development. Although some studies on sexual reproduction have been carried out in model plants, little is known about the dynamic changes in DNA methylation and their effect on gene expression in sexual and asexual progeny of woody plants. Here, through whole-genome bisulfite sequencing, we revealed DNA methylation patterns in the sexual and asexual progenies of mature Robinia pseudoacacia to understand the regulation of gene expression by DNA methylation in juvenile seedlings. An average of 53% CG, 34% CHG and 5% CHH contexts was methylated in the leaves of mature and juvenile individuals. The CHH methylation level of asexually propagated seedlings was significantly lower than that of seed-derived seedlings and mature trees. The intergenic regions had the highest methylation level. Analysis of differentially methylated regions (DMRs) showed that most of them were hypermethylated and located in the gene upstream and introns. A total of 24, 108 and 162 differentially expressed genes containing DMRs were identified in root sprouts (RSs), root cuttings (RCs) and seed-derived seedlings (SSs), respectively, and a large proportion of them showed hypermethylation. In addition, DMRs were enriched within GO subcategories including catalytic activity, metabolic process and cellular process. The results reveal widespread DNA methylation changes between mature plants and their progenies through sexual/asexual reproduction, which provides novel insights into DNA methylation reprogramming and the regulation of gene expression in woody plants.

Shikonin ameliorates injury and inflammatory response of LPS-stimulated WI-38 cells via modulating the miR-489-3p/MAP2K1 axis.

Pneumonia is an inflammatory disease induced by infection with different pathogens. Currently, multiple preclinical studies have revealed that shikonin, a natural naphthoquinone, can mitigate lipopolysaccharide (LPS)-induced inflammation, but its underlying mechanism in pneumonia remains unknown. This research was designed to explore the function and regulatory mechanism of shikonin in LPS-induced cell injury and inflammation in WI-38 cells. In-vitro model of pneumonia was constructed by treating WI-38 cells with LPS. Expression of miR-489-3p and MAP2K1 was tested by RT-qPCR and (or) Western blot analysis. Cell viability was examined by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay. The productions of pro-inflammatory cytokines were determined by enzyme-linked immunosorbent assays. Cell apoptosis was detected by Western blot and flow cytometry analysis. In the current study, LPS induced WI-38 cell damage by inhibiting cell viability and promoting cell apoptosis and inflammation. Shikonin ameliorated LPS-induced cell injury and elevated miR-489-3p expression. LPS-induced inflammatory injury was further mitigated by upregulation of miR-489-3p. In addition, MAP2K1, the target of miR-489-3p, was upregulated by LPS. Furthermore, upregulation of MAP2K1 reversed the influence of shikonin and miR-489-3p mimics on LPS-induced cell injury and inflammation. This study revealed that shikonin protected WI-38 cells against LPS-induced cell injury and inflammatory response by regulating the miR-489-3p/MAP2K1 axis, thus affecting the progression of pneumonia.

Influence of Serum Albumin on HbA1c and HbA1c-Defined Glycemic Status: A Retrospective Study.

Background: Glycated hemoglobin (HbA1c) is commonly used in the diagnosis and evaluation of glycemic control in diabetes, and it may be influenced by several non-glycemic and glycemic factors, including albumin. This retrospective study investigated the influence of albumin on HbA1c and HbA1c-defined glycemic status. Methods: The demographic, hematological, and biochemical data were collected for 11,922 patients undergoing routine physical examination. Univariate and multivariate linear regression analyses, stratified analyses and interaction analyses, and multiple logistic regression were conducted to identify the association between albumin and HbA1c in people with different glycemic status. Results: HbA1c levels were inversely associated with serum albumin level (P < 0.0001) in all participants. Risk factors leading to the association included age > 45 years, high fasting plasma glucose (≥7.0 mmol/L), and anemia. The negative association between HbA1c and albumin was curved (P < 0.0001) and had a threshold effect in the HbA1c-defined diabetic population; the association was significantly stronger when the albumin level fell below 41.4 g/L (ß: -0.31, 95% CI: -0.45 to -0.17, P < 0.0001). A 2 g/L increase in albumin reduced the odds of HbA1c-defined dysglycemia, diabetes, and poor glycemia control by 12% to 36%, after adjustment for all possible confounders. Conclusions: HbA1c was inversely associated with albumin level in all participants, and the association was significantly stronger in people with diabetes (defined by HbA1c criteria). For diabetic patients with lower albumin level, there was an increased risk of an erroneous HbA1c-based identification and management of glycemic status.

Bioinformatic prediction of immunodominant regions in spike protein for early diagnosis of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

BACKGROUND: To contain the pandemics caused by SARS-CoV-2, early detection approaches with high accuracy and accessibility are critical. Generating an antigen-capture based detection system would be an ideal strategy complementing the current methods based on nucleic acids and antibody detection. The spike protein is found on the outside of virus particles and appropriate for antigen detection. METHODS: In this study, we utilized bioinformatics approaches to explore the immunodominant fragments on spike protein of SARS-CoV-2. RESULTS: The S1 subunit of spike protein was identified with higher sequence specificity. Three immunodominant fragments, Spike56-94, Spike199-264, and Spike577-612, located at the S1 subunit were finally selected via bioinformatics analysis. The glycosylation sites and high-frequency mutation sites on spike protein were circumvented in the antigen design. All the identified fragments present qualified antigenicity, hydrophilicity, and surface accessibility. A recombinant antigen with a length of 194 amino acids (aa) consisting of the selected immunodominant fragments as well as a universal Th epitope was finally constructed. CONCLUSION: The recombinant peptide encoded by the construct contains multiple immunodominant epitopes, which is expected to stimulate a strong immune response in mice and generate qualified antibodies for SARS-CoV-2 detection.

Circulating levels of IL-33 are elevated by obesity and positively correlated with metabolic disorders in Chinese adults.

BACKGROUND: Interleukin-33 (IL-33) plays a pivotal role in regulating innate immune response and metabolic homeostasis. However, whether its circulating level is correlated with obesity and metabolic disorders in humans remains largely unknown. We aimed to address this gap by determining IL-33 serum level and its downstream type 2 inflammatory cytokines interleukin-5 (IL-5) and interleukin-13 (IL-13) in overweight/obese population, and analyzing the specific associations between IL-33 and obesity metabolic phenotypes. METHODS: 217 subjects were enrolled and divided into three groups: healthy control (HC) subjects, metabolically healthy overweight/obese (MHOO) subjects and metabolically unhealthy overweight/obese (MUOO) subjects. Circulating levels of IL-33, IL-5 and IL-13 were measured using ELISA analyses. Multivariate regression analyses were further performed to determine the independent association between IL-33 and obesity metabolic phenotypes. RESULTS: Circulating levels of IL-33 were significantly elevated in subjects of MUOO group compared with HC group and MHOO group, while no significant difference was observed between the latter two groups in IL-33 levels. Consistent with this, serum levels of IL-5/13 were higher in the MUOO group compared with HC and MHOO groups. After adjusted for all confounders, MUOO phenotype was significantly associated with increased IL-33 serum levels (OR = 1.70; 95% CI 1.09-2.64; p = 0.019). With the MHOO group as the reference population, higher circulating level of IL-33 was also positively associated with MUOO phenotype after adjusting for confounders (OR = 1.50; 95% CI 1.20-1.88; p = 2.91E-4). However, there was no significant association between MHOO phenotype and IL-33 levels (p = 0.942). Trend analysis further confirmed the positive correlation between MUOO phenotype and IL-33 level (p for trend = 0.019). Additionally, IL-33 was significantly and positively correlated with diastolic blood pressure (DBP), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), white blood cell (WBC), neutrophil and IL-5 only in MUOO group, while inversely correlated with high density lipoprotein cholesterol (HDL-C) in MHOO subjects. CONCLUSION: Circulating levels of IL-33 were significantly elevated in overweight/obese Chinese adults with metabolic disorders. Increased levels of IL-33 were positively associated with metabolically unhealthy overweight/obese phenotype and several metabolic syndrome risk factors.

A Novel Cleavage Pattern of Complement C5 Induced by Chlamydia trachomatis Infection via the Chlamydial Protease CPAF.

Urogenital Chlamydia trachomatis infection is one of the most common bacterial sexually transmitted diseases globally. Untreated C. trachomatis infections can ascend to the upper genital tract and establish a series of severe complications. Previous studies using C3-/- and C5-/- mice models demonstrated that C3-independent activation of C5 occurred during C. trachomatis infection. However, the mechanism of how chlamydial infection activates C5 in the absence of C3 has yet to be elucidated. To delineate interactions between C5 and chlamydial infection, cleavage products in a co-incubation system containing purified human C5 and C. trachomatis-HeLa229 cell lysates were analyzed, and a novel cleavage pattern of C5 activation induced by C. trachomatis infection was identified. C5 was cleaved efficiently at the previously unidentified site K970, but was cleaved poorly at site R751. C5b was modified to C5bCt, which later formed C5bCt-9, which had enhanced lytic ability compared with C5b-9. The chlamydial serine protease CPAF contributed to C3-independent C5 activation during C. trachomatis infection. Nafamostat mesylate, a serine protease inhibitor with a good safety profile, had a strong inhibitory effect on C5 activation induced by chlamydial infection. These discoveries reveal the mechanism of C3-independent C5 activation induced by chlamydial infection, and furthermore provide a potential therapeutic target and drug for preventing tubal fibrosis caused by chlamydial infection.

Omega-3 fatty acid supplementation is associated with favorable outcomes in patients with sepsis: an updated meta-analysis.

OBJECTIVES: The efficacy of omega-3 fatty acids in the treatment of sepsis is controversial. We conducted an updated meta-analysis to clarify the efficacy of omega-3 fatty acids in patients with sepsis. METHODS: PubMed, EMBASE, and the Cochrane Library were searched for randomized clinical trials (RCTs) on omega-3 fatty acid supplementation in adults with sepsis. RESULTS: Twenty eligible RCTs involving 1514 patients were included in the meta-analysis. Omega-3 fatty acid supplementation was linked to reductions of mortality (I2 = 0, relative risk [RR] = 0.82, 95% confidence interval [CI] = 0.69-0.97), the duration of mechanical ventilation (DMV; I2 = 74%, weighted mean difference [WMD] = -2.20, 95% CI = -4.00 to -0.40), and intensive care unit (ICU) length of stay (LOS; I2 = 91%, WMD = -3.86, 95% CI = -5.72 to -2.01). Subgroup analysis illustrated that mortality was significantly reduced in patients with sepsis and gastrointestinal dysfunction (RR = 0.5, 95% CI = 0.29-0.86, I2 = 0). CONCLUSION: Omega-3 fatty acid supplementation might be associated with reduced mortality in patients with sepsis, especially those with gastrointestinal dysfunction. Furthermore, omega-3 fatty acid administration could shorten DMV and ICU LOS.

Delirium During Recovery in Patients With Severe COVID-19: Two Case Reports.

Novel coronavirus disease (COVID-19), caused by SARS-CoV-2, has rapidly evolved into a worldwide pandemic, leaving patients with life-threatening respiratory, cardiovascular, and cerebral complications. Here we reported on two patients with severe COVID-19 who experienced delirium in the early stage of recovery and mental illness including fatigue, anxiety, and post-traumatic stress disorder in the post-illness stage of COVID-19. Two patients were admitted to hospital due to clinical symptoms and features of CT and were confirmed for COVID-19 by positive results of a throat swab for SARS-CoV-2. Due to severe respiratory symptoms and a low oxygenation index, they were transferred to the ICU and received invasive mechanical ventilation and sedation. Hyperactive delirium was observed after being transferred out of the ICU. Different treatment measures were taken in time. Delirium did not occur again in hospital, but they showed mental suffering, including fatigue, anxiety, and post-traumatic stress disorder (PTSD), during the 5 month follow-up after discharge.

Immunodominant regions prediction of nucleocapsid protein for SARS-CoV-2 early diagnosis: a bioinformatics and immunoinformatics study.

COVID-19 caused by SARS-CoV-2 is sweeping the world and posing serious health problems. Rapid and accurate detection along with timely isolation is the key to control the epidemic. Nucleic acid test and antibody-detection have been applied in the diagnosis of COVID-19, while both have their limitations. Comparatively, direct detection of viral antigens in clinical specimens is highly valuable for the early diagnosis of SARS-CoV-2. The nucleocapsid (N) protein is one of the predominantly expressed proteins with high immunogenicity during the early stages of infection. Here, we applied multiple bioinformatics servers to forecast the potential immunodominant regions derived from the N protein of SARS-CoV-2. Since the high homology of N protein between SARS-CoV-2 and SARS-CoV, we attempted to leverage existing SARS-CoV immunological studies to develop SARS-CoV-2 diagnostic antibodies. Finally, N229-269, N349-399, and N405-419 were predicted to be the potential immunodominant regions, which contain both predicted linear B-cell epitopes and murine MHC class II binding epitopes. These three regions exhibited good surface accessibility and hydrophilicity. All were forecasted to be non-allergen and non-toxic. The final construct was built based on the bioinformatics analysis, which could help to develop an antigen-capture system for the early diagnosis of SARS-CoV-2.

High expression of miR-483-5p aggravates sepsis-induced acute lung injury.

Sepsis-induced acute lung injury (ALI) has high morbidity and mortality rates, and there remains a need for therapeutic methods to improve the outcome of ALI patients. miR-483-5p is an important regulator for the development of various diseases such as sepsis. Nevertheless, it is not known whether miR-483-5p has an effect on sepsis-induced ALI. To explore this issue, this study used cecal ligation and puncture (CLP)-treated mice and lipopolysaccharide (LPS)-treated pulmonary microvascular endothelial cells (PMVECs) cells to simulate the models of sepsis-induced ALI in vivo and in vitro. Pathological and histological changes of lungs from sepsis-induced ALI mice were detected by Hematoxylin-eosin staining. The detection levels of caspase-3, interleukin (IL)-6 and IL-1ß were used to reflect the effect of miR-483-5p on apoptosis and inflammation of sepsis-induced ALI. The detection level of lactate dehydrogenase (LDH) in PMVECs cells was used to reflect the severe extent of sepsis-induced injury. The expression of miR-483-5p in lung tissues of sepsis-induced ALI mice was determined by qRT-PCR. In addition, the interaction of miR-483-5p with PIAS1 was identified and validated by Targetscan website and luciferase reporter assay, respectively. The results showed that miR-483-5p was up-regulated in the lung tissues of sepsis-induced ALI mice. Knockdown of miR-483-5p effectively ameliorated lung injury in mice with sepsis-induced ALI and inhibited inflammation and apoptosis of LPS-treated PMVECs cells. Furthermore, in vitro experiment revealed that PIAS1 was a potential target of miR-483-5p. Moreover, miR-483-5p could suppress PIAS1 expression to aggravate inflammation and apoptosis of LPS-treated PMVECs cells. These findings suggest miR-483-5p is a potential therapeutic and diagnostic biomarker for sepsis-induced ALI and provide a new insight for understanding the molecular mechanism of sepsis-induced ALI.

[Prenatal diagnosis for two fetuses carrying partial deletion of Y chromosome].

OBJECTIVE: To perform prenatal diagosis for two fetuses carrying partial deletion of Y chromosome. METHODS: Routine G- and C-banding were carried out to analyze the chromosomal karyotypes of the fetuses and their fathers. Fetal DNA was also subjected to low-coverage massively parallel copy number variation sequencing (CNV-seq), fluorescence in situ hybridization (FISH), SRY gene and AZF factor testing. RESULTS: Both fetuses showed a 46, XN, del(Y) (q11.2) karyotype at 320-400 band level by the analysis of amniotic fluid chromosomes. FISH with Y chromosome centromere probe indicated that in both cases the number of Y chromosome was normal. Both fathers had an apparently normal karyotype at 320-400 band level. For fetus 1, CNV-seq test revealed a 12.88 Mb deletion at Yq11.221-q12, which encompassed the whole of AZFb+AZFc regions and may lead to male infertility, sperm deficiency and/or severe oligospermia. In fetus 2, CNV-seq also detected a 14.84 Mb deletion at Yq11.21-q12, which encompassed the whole of the AZF region and may lead to severe spermatogenesis disorder resulting in severe oligoasthenospermia and azoospermia. In both cases, testing of SRY gene was positive. No point mutation of the SRY gene was identified. Analysis of amniotic fluid DNA confirmed partial or total absence of AZF in the two fetuses, respectively. CONCLUSION: Combined use of various technologies can enable accurate detection of structural abnormalities of the Y chromosome and facilitate genetic counseling. CNV-seq can help with rapid screening of Y chromosome microdeletions and may be used as a complementary test for chromosomal karyotyping.

Me6-TREN/TREN Mixed-Ligand Effect During SET-LRP in the Catalytically Active DMSO Revitalizes TREN into an Excellent Ligand.

A mixed-ligand effect was observed for mixtures of tris(2-dimethylaminoethyl)amine (Me6-TREN) with tris(2-aminoethyl)amine (TREN) ligands during Cu(0) wire-catalyzed, single-electron transfer-living radical polymerization (SET-LRP) of methyl acrylate (MA) initiated with bis(2-bromopropionyl)ethane (BPE) in DMSO. The external order of reaction of SET-LRP both in the presence of Me6-TREN, TREN and of the mixed-ligand Me6-TREN/TREN, in DMSO, demonstrated a catalytic activity for DMSO similar to that reported in the presence of Cu(0) powder. The catalytic activity of DMSO, with close to 100% chain-end functionality, facilitates the much less expensive TREN to act as a very efficient ligand that is competitive with Me6-TREN and with the mixed-ligand and revitalizes TREN into an excellent ligand. The highest activity of the mixed-ligand at 1/1 ratio between ligands suggests that in addition to a fast exchange between these two ligands, a new single dynamic ligand stabilized by hydrogen-bonding, may generate these results.

Real-time PCR assay may be used to verify suspicious test results of Ureaplasmas spp. from the liquid culture method.

Ureaplasma spp. are associated with female genital tract infections and are mainly tested by liquid culture in developing countries. To evaluate the accuracy of liquid culture, 686 vaginal swabs were collected and tested by using the Mycoplasma Culturing, Identification, Enumeration, and Susceptibility (IES) Kit. Then these culture broths were verified using real-time PCR. Among 368 Ureaplasma positive broths, 263 contained Ureaplasma parvum, 30 contained Ureaplasma urealyticum, 57 contained both, and 18 were negative by real-time PCR. In 318 Ureaplasmas negative broths, 78 were found to be Ureaplasma positive by real-time PCR. Using real-time PCR as the reference, the false positive rate of the liquid culture was 7.0%. It has been suggested that the liquid culture positive broth should be inoculated onto solid agar to eliminate false-positives. However, solid culture is rarely used due to low sensitivity and being time consuming. Real-time PCR may be performed to replace solid culture to verify suspicious liquid culture results.

Replacing Cu(II)Br2 with Me6-TREN in Biphasic Cu(0)/TREN Catalyzed SET-LRP Reveals the Mixed-Ligand Effect.

The mixed-ligand system consisting of tris(2-aminoethyl)amine (TREN) and tris(2-dimethylaminoethyl)amine (Me6-TREN) during the Cu(0) wire-catalyzed single electron transfer-living radical polymerization (SET-LRP) of methyl acrylate (MA) in "programmed" biphasic mixtures of the dipolar aprotic solvents NMP, DMF, and DMAc with H2O is reported. Kinetic and chain end analysis studies by NMR and MALDI-TOF before and after thio-bromo "click" reaction demonstrated that Me6-TREN complements and makes the less expensive TREN a very efficient ligand in the absence of externally added Cu(II)Br2. Statistical analysis of the kinetic data together with control experiments demonstrated that this mixed-ligand effect enhanced the apparent rate constant of propagation, monomer conversion, and molecular weight control. The most efficient effect was observed at a 1/1 molar ratio between these two ligands, suggesting that in addition to a fast exchange between the two ligands, a new single dynamic ligand generated by hydrogen bonding may be responsible for the mixed ligand observed.

[Genetic analysis and prenatal diagnosis of a fetus with harlequin ichthyosis].

OBJECTIVE: To carry out variant analysis for a fetus suspected with harlequin ichthyosis (HI). METHODS: Whole exome sequencing (WES) was employed to detect potential variant in the fetus. Suspected variant was validated by Sanger sequencing. RESULTS: A homozygous missense variant c.6858delT (p.F2286fs) was detected in the fetus, for which both parents were heterozygous carriers. Pathological analysis confirmed the diagnosis of HI. CONCLUSION: The c.6858delT variant of the ABCA12 gene probably underlies the disease in the fetus.