Frozen Translational and Rotational Motion of Human Immunodeficiency Virus Transacting Activator of Transcription Peptide-Modified Nanocargo on Neutral Lipid Bilayer

With time-resolved high-precision single-particle tracking methodologies, we explored the adsorption and thermal motion of transacting activator of transcription (TAT) peptide-modified nanocargo on a model lipid bilayer in the nonelectrostatic domain. We found that the lateral and rotational motion of TAT peptide-modified nanocargo could be effectively suppressed on the surface of neutral lipid membrane, a feature that cannot be explained by existing hypotheses. A semiquantitative association activation energy analysis revealed that multiple weak bonds were required for the initial adsorption process. As a result, the localized multiple TAT peptides on the surface of the nanocargo can provide a pathway for the creation of a net of peptide–lipid complexes (e.g., lipid domain). The dragging forces caused by these complexes effectively confined the thermal motion of the nanocargo on the fluid membrane that cannot be achieved by individual peptides with random spatial and conformational distributions. These interesting findings could provide insightful information for the better understanding of the intracellular internalization mechanism of TAT peptide-modified nanocargo and might shed new light on the development of highly efficient intracellular carriers for site-specific delivery of drugs and genes

Probing Single-Molecule Fluorescence Spectral Modulation within Individual Hotspots with Subdiffraction-Limit Image Resolution

The enhancement of the electromagnetic field on the rough metallic nanostructure has been extensively applied to obtain chemical or biological information about molecules with high sensitivity and has received much attention due to its potential applications in new types of devices based on nanoelectronics and nanophotonics. The typical size of the field enhancement area, the so-called hotspot, is approximately 1 order of magnitude smaller than the optical diffraction limit. In the present study, an optical super-resolution microscopic and spectroscopic approach is introduced to explore single-molecule fluorescence within a hotspot where nonhomogeneous spectral modulation is resolved beyond the optical diffraction limit for the first time. Distinct Stokes shifts from individual dyes were directly observed within single hotspots, which were found to be independent of the local electromagnetic field strength. The method reported here provides a robust tool to probe the optical properties of nanoresonantors with high temporal and spatial resolution

iPS progenitor cells vs. MEFs and feature correlation.

(a) shows the examples of iPS progenitor cell images (blue circles) and normal MEFs images (yellow boxes) taken from phase 1, 2 and 3 of field 2 (Left, middle and right). Nucleus and cytoplasm of the enlarged progenitor cells and normal MEFs are colored in light blue and green respectively. (b) shows the Pearson coefficients between remaining types of features in three phases after the first step of feature selection. Note in this figure ellipsoid-prolate is denoted as E-prolate, intensity-StdDev as I-stdDev, intensity-min as I-Min, intensity-max as I-Max, nucleus-cytoplasm volume ratio as Ratio, ellipsoid-oblate as E-oblate.</p

Feature ranking and selection.

This figure shows how the precision values change with the deleted feature in a recursive fashion. Least important features are removed earlier.</p

Time window selection.

The three subplots represent the precision values for different time windows based on 21 start frames (x axis) and 12 window lengths (7 frames to 29 frames) for phases 1, 2, and 3 (from top to bottom) respectively, and the black bash line in each subplot indicates a precision value of 0.55.</p

Loss of <i>Gsα</i> in NCCs results in severe craniofacial malformations.

<p>(A, B) P0 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutant and control. <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutant mice become cyanotic and die within hours after birth, exhibit domed skull, shortened maxilla and mandible and exposed tongue. (C-F) The gross appearance of <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutants (C and E) and controls (D and F) are identical at E10.5 and E12.5 respectively. (G, H) E14.5 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutant and control. <i>Wnt1-cre;Gsα</i>^<i>f/f</i> embryos show short snout, round face and hypertelorism. (I-L) E16.5 and E18.5 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutants (I and K) display exposed tongues and shortened maxilla and mandible compare to controls (J and L).</p

Loss of <i>Gsα</i> in NCCs results in abnormal ossification.

<p>(A-H) Von Kossa and nuclear red staining of heads coronal sections in <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutants and controls from different embryonic stages. The aggregated mesenchymal cell in maxilla are similar between E12.5 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutant and control (arrows in A and B); however, the ossification region in E14.5 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutant is larger than that in control (arrows in C and D), and this phenotype are much more severe at later embryonic stages (arrows in E-H). (I-L) Von Kossa staining (I, J) and alcian blue staining (K, L) show the abnormal ossification and malformation of nasal septum cartilage in E17.5 <i>Wnt1-cre;Gsα</i>^<i>f/f</i> mutants (arrows in I and K).</p

Model validation.

In all sub-figures, X axis indicates the start frame of the best time windows and the corresponding window length (13 frames) is indicated in the inlet. (a) 5-fold cross-validation precisions over 10 runs. (b) the standard deviation of the average precision of the neighborhood time windows in Fig 6D. (c) the standard deviation of the average precision of the distant windows in Fig 6E. (d) the average precision of seven neighborhood time windows calculated over 10 holdout validation runs. (e) the average precision over 10 independent tests for six best time windows on their corresponding distant windows.</p

Model comparison for different missing frame number and imputation methods.

(a) shows the average precision over six time periods (TP1 to TP6) for each missing frame number and imputation method set_KNN (colored as blue), set_mean (colored as red), set_mean_mod (colored as green) and all three imputation methods (colored as gray). (b) shows the standard deviation, as a function of missing frame number, of imputation method set_KNN (colored as blue), set_mean (colored as red), set_mean_mod (colored as green) and all three imputation methods (colored as gray).</p

DataSheet_1_Non-Assisted Hatching Trophectoderm Biopsy Does Not Increase The Risks of Most Adverse Maternal and Neonatal Outcome and May Be More Practical for Busy Clinics: Evidence From China.docx

ObjectiveThis study was conducted in order to investigate whether non-assisted hatching trophectoderm (TE) biopsy increases the risks of adverse perinatal outcomes in livebirths following elective single cryopreserved-thawed blastocyst transfer.Patients and MethodsA total of 5,412 cycles from 4,908 women who achieved singleton livebirths between 2013 and 2019 were included in this retrospective cohort study. All embryos in this study were fertilized by intracytoplasmic sperm injection (ICSI) and cryopreserved through vitrification. The main intervention is to open the zona pellucida (ZP) of day 5/6 blastocyst immediately for biopsy without pre-assisted hatching. The main outcome measures are the common maternal and neonatal outcomes, including hypertensive disorders of pregnancy (HDPs), gestational diabetes mellitus (GDM), abnormal placentation, abnormalities in umbilical cord and amniotic fluid, preterm birth, cesarean section, low birth weight, postpartum hemorrhage, and prolonged hospital stay (both mothers and infants). The generalized estimation equation (GEE) was used to control the effects of repeated measurements. The non-conditional logistic regression model was used to examine the associations between embryo biopsy status and each adverse perinatal event. Given that the selection bias and changes in learning curve might affect the results, we selected 1,086 similar (matching tolerance = 0.01) cycles from the ICSI group via propensity score matching (PSM) for second comparisons and adjustment (conditional logistic regression).ResultsAfter adjusting for confounders, we confirmed that the non-assisted hatching protocol did not increase the risks of most adverse maternal and neonatal outcomes. Despite this, there were increased risks of GDM (aOR: 1.522, 95% CI: 1.141–2.031) and umbilical cord abnormalities (aOR: 11.539, 95% CI: 1.199–111.067) in the biopsy group. In the second comparisons after PSM, GDM incidence in the biopsy group was still higher (7.26% vs. 5.16%, P = 0.042), yet all measurement outcomes were equally likely to occur in both groups after the second adjustment.ConclusionsThe non-assisted hatching TE biopsy does not increase the risks of most adverse perinatal outcomes. However, there is a higher GDM incidence in the biopsy group, and this association warrants further study. Considering its safety and simplicity, the non-assisted hatching protocol has the potential to become the preferred option for TE biopsy, especially in busy clinics and IVF laboratories.</p