Accurate SNV detection in single cells by transposon-based whole-genome amplification of complementary strands.

Single-nucleotide variants (SNVs), pertinent to aging and disease, occur sporadically in the human genome, hence necessitating single-cell measurements. However, detection of single-cell SNVs suffers from false positives (FPs) due to intracellular single-stranded DNA damage and the process of whole-genome amplification (WGA). Here, we report a single-cell WGA method termed multiplexed end-tagging amplification of complementary strands (META-CS), which eliminates nearly all FPs by virtue of DNA complementarity, and achieved the highest accuracy thus far. We validated META-CS by sequencing kindred cells and human sperm, and applied it to other human tissues. Investigation of mature single human neurons revealed increasing SNVs with age and potentially unrepaired strand-specific oxidative guanine damage. We determined SNV frequencies along the genome in differentiated single human blood cells, and identified cell type-dependent mutational patterns for major types of lymphocytes.

Biased migration of confined neutrophil-like cells in asymmetric hydraulic environments.

Cells integrate multiple measurement modalities to navigate their environment. Soluble and substrate-bound chemical gradients and physical cues have all been shown to influence cell orientation and migration. Here we investigate the role of asymmetric hydraulic pressure in directional sensing. Cells confined in microchannels identified and chose a path of lower hydraulic resistance in the absence of chemical cues. In a bifurcating channel with asymmetric hydraulic resistances, this choice was preceded by the elaboration of two leading edges with a faster extension rate along the lower resistance channel. Retraction of the "losing" edge appeared to precipitate a final choice of direction. The pressure differences altering leading edge protrusion rates were small, suggesting weak force generation by leading edges. The response to the physical asymmetry was able to override a dynamically generated chemical cue. Motile cells may use this bias as a result of hydraulic resistance, or "barotaxis," in concert with chemotaxis to navigate complex environments.

Comparing outcomes of ovarian cystectomy by vaginal natural orifice transluminal endoscopic surgery versus laparoendoscopic single-site surgery: A retrospective cohort study.

BACKGROUND: We aimed to compare the outcomes of ovarian cystectomy (OC) performed by vaginal natural orifice transluminal endoscopic surgery (vNOTES) vs transumbilical laparoendoscopic single-site surgery (LESS). METHODS: We retrospectively analyzed the data of patients in our hospital who underwent OC either by vNOTES or LESS between January 2015 and September 2021. Demographic data were collected. The primary outcome was the conversion rate. The secondary outcomes were the duration of surgery, length of hospital stay, estimated blood loss, maximum body temperature within 48 hours after operation, and duration of maximum body temperature (hours), among others. Statistical analysis was done using the SPSS software. RESULTS: Exactly 284 patients were screened. The vNOTES and LESS groups consisted of 21 and 47 patients, respectively. There was no significant difference in the conversion rates between the two groups (0 vs 8.5% in vNOTES and LESS, respectively; p = 0.303). Compared with the vNOTES group, the LESS group had a larger cyst diameter (6.00 ± 2.32 vs 4.69 ± 1.29 cm; p = 0.004), more endometriotic cysts (42.6% vs 9.5%; p < 0.001), and more pelvic adhesions requiring adhesiolysis (57.4% vs 19.0%; p = 0.003). At baseline, there were no other differences between the groups. The secondary outcomes included a shorter duration of surgery (70.14 ± 27.30 vs 99.57 ± 36.26 minutes; p = 0.001) and lower estimated blood loss (64.29 ± 39.19 vs 163.43 ± 251.20 mL; p = 0.011) in the vNOTES group. Regression analysis showed the diameter of the ovarian cyst correlated with surgical time. The complication was comparable between the two groups. CONCLUSION: Above all, the advantages of vNOTES include an absence of visible scars, shorter surgical duration, and less blood loss when compared with LESS. Further large-scale prospective trials should confirm the results of our study.

Transient ischemic attack after mRNA-based COVID-19 vaccination during pregnancy: A case report.

BACKGROUND: Thrombocytopenia with thrombosis syndrome has been reported after vaccination against severe acute respiratory syndrome coronavirus 2 with two mRNA vaccines. The syndrome is characterized by thrombosis, especially cerebral venous sinus thrombosis, and may lead to stroke. Pregnant women with stroke show higher rates of pregnancy loss and experience serious pregnancy complications. We present the case of a 24-year-old pregnant woman with a transient ischemic attack (TIA) that developed after vaccination with the Moderna mRNA-1273 vaccine (at 37 2/7 wk). CASE SUMMARY: TIA occurred 13 d following the coronavirus disease vaccination. At 39 1/7 wk of pregnancy, the patient presented with sudden onset of right eye blurred vision with headache, dizziness with nausea, right-hand weakness, anomia, and alexia. The symptoms lasted 3 h; TIA was diagnosed. Blood test results revealed elevated D-dimer, cholesterol, and triglyceride levels. Brain magnetic resonance imaging showed no acute hemorrhagic or ischemic stroke. At pregnancy 37 6/7 wk, she was admitted for cesarean delivery to reduce subsequent risk of stroke during labor. Body mass index on admission was 19.8 kg/m2. Magnetic resonance angiography and transesophageal echocardiography showed no abnormalities. The next day, a mature female baby weighing 2895 g and measuring 50 cm was delivered. Apgar scores were 8 and 9 in the first and fifth minutes. D-dimer levels decreased on postoperative day 4. After discharge, the autoimmune panel was within normal limits, including antinuclear and antiphospholipid antibodies. CONCLUSION: TIA might be developed after the mRNA vaccines in pregnant women.

Vulvar myoma: A case report and review of literature.

OBJECTIVE: To present a case of vulvar myoma and the factors differentiating this tumor from Bartholin's cyst. CASE REPORT: A 50-year-old woman presented with a nodule over the left labia majora. Pelvic examination showed swelling and redness of the left labia majora. A 2-cm nodule with firm consistency was found near the vaginal opening. There was no inguinal lymphadenopathy. Bartholin's cyst was suggested, and oral cephalexin was prescribed for 1 week, but no improvement was seen. Therefore, she underwent excision of the nodule. Pathology revealed it to be a benign vulvar myoma. The patient recovered well, and no recurrence was noted after 2 months of follow-up. CONCLUSION: Vulvar myoma is rare. Sexual history, nodule consistency, and imaging are helpful, but the final diagnosis of vulvar myoma is usually made following surgical excision and histopathological analysis.

Risk of chronic spontaneous urticaria in reproductive-aged women with abnormal uterine bleeding: A population-based cohort study.

Women with abnormal uterine bleeding (AUB) have been reported to develop chronic spontaneous urticaria (CSU). Nevertheless, whether or not AUB women have an increased risk of CSU has not been examined in large-scale epidemiologic studies. This study aimed to investigate the risk of CSU among reproductive-aged women with AUB. A total of 79 595 patients and 79 107 propensity-score matched controls were recruited from Taiwan's National Health Insurance Research Database to conduct a nationwide cohort study. The Cox proportional-hazard regression model was applied to examine the adjusted hazard ratios (aHR) and 95% confidence intervals (CI) for CSU in relation to AUB. We found that women with AUB had a higher risk for CSU (aHR = 1.83; 95% CI, 1.76-1.90) than women without AUB. Subgroup analyses revealed that AUB with an abnormal bleeding frequency (aHR = 1.70; 95% CI, 1.60-1.79), irregular bleeding (aHR = 1.80; 95% CI, 1.71-1.89), and intermenstrual bleeding (aHR = 1.65; 95% CI, 1.49-1.83) were associated with an increased risk of CSU compared with those without abnormalities. The Kaplan-Meier analysis revealed that the cumulative incidence of developing CSU was consistently higher in the AUB cohort than in the non-AUB cohort during the entire follow-up period (log-rank test, p < 0.001). In conclusion, reproductive-aged women with AUB were found to have a higher risk of developing CSU. This study emphasizes the importance of enquiring CSU patients about menstrual problems in clinical practice. Further consultation with obstetrician-gynecologists may be beneficial.

Three-dimensional genome structures of single diploid human cells.

Three-dimensional genome structures play a key role in gene regulation and cell functions. Characterization of genome structures necessitates single-cell measurements. This has been achieved for haploid cells but has remained a challenge for diploid cells. We developed a single-cell chromatin conformation capture method, termed Dip-C, that combines a transposon-based whole-genome amplification method to detect many chromatin contacts, called META (multiplex end-tagging amplification), and an algorithm to impute the two chromosome haplotypes linked by each contact. We reconstructed the genome structures of single diploid human cells from a lymphoblastoid cell line and from primary blood cells with high spatial resolution, locating specific single-nucleotide and copy number variations in the nucleus. The two alleles of imprinted loci and the two X chromosomes were structurally different. Cells of different types displayed statistically distinct genome structures. Such structural cell typing is crucial for understanding cell functions.

Shrinkage behaviors of dental composite restorations-The experimental-numerical hybrid analysis.

OBJECTIVES: To investigate the effects of light curing protocols on the shrinkage behaviors, contraction stress, and microleakage in composite restorations by an experimental-numerical hybrid analysis. METHODS: Three groups of human molars were collected to receive different light-curing protocols: vertical or oblique curing at regular intensity, and vertical curing at reduced intensity. For each tooth, the composite fillings were consecutively placed under unbonded and bonded states, and their shrinkage behaviors were examined with a digital image correlation (DIC) technique. The strains of the unbonded restorations were input into two finite element analysis (FEA) models with settings of the composite as either homogeneous or hardened along polymerization gradients. The preliminary solutions were verified by their individual deformations in the bonded restorations. The interfacial microleakage of restorations was also determined by micro-CT scanning and compared with the FEA results. RESULTS: The bonded restorations showed centripetal shrinkage patterns with greater downward displacements than their unbonded restorations. Vertical curing at regular intensity caused the greatest shrinkage strain, contraction stress, and microleakage among the three protocols. Low-intensity curing reduced overall shrinkage strain and displacements at cervical margin, but did not prevent the formation of microleakage. Oblique curing caused asymmetric shrinkage with the tooth-shielded side revealing less deformation. Setting the polymerization-dependent elastic moduli of the composite enhanced the reliability of FEA. SIGNIFICANCE: This hybrid analysis comprehensively examined the polymerization shrinkage behaviors. Both the light intensity and direction affect the shrinkages and contraction stress. Oblique curing decreases shrinkage due to the attenuated irradiation by tooth-shielding rather than modulations of shrinkage direction.

Early reperfusion improves the recovery of contralateral electrophysiological diaschisis following focal cerebral ischemia in rats.

OBJECTIVES: We evaluated electrophysiological benefits of reperfusion following ischemic stroke. METHODS: Rats received either transient proximal occlusion of the right middle cerebral artery for 30 (Group I, n=8) or 90 minutes (Group II, n=8) or permanent thermocoagulation of the distal right middle cerebral artery (Group III, n=6). Neurobehavioral outcome and somatosensory evoked potentials (SSEPs) were examined before and at 7 days after the onset of brain ischemia. Brain infarction was assessed after the rats were euthanized. RESULTS: Before ischemia, stable SSEPs were consistently recorded. At 7 days post-insult, Group III (permanent occlusion) had the greatest reduction in the SSEPs recorded ipsilaterally and contralaterally. Groups I and II (transient ischemic groups) also had depressant SSEPs recorded from the ipsilateral ischemic and the contralateral intact brain (electrophysiological diaschisis). However, prolonged ischemia resulted in greater brain infarction and increased neurological deficits in addition to greater reductions in the ipsilateral and the contralateral SSEPs. CONCLUSION: Early reperfusion facilitates the electrophysiological recovery in both ipsilateral lesional and the contralateral intact brain, which may be closely relevant to post-injury brain rewiring. We also demonstrated that contralateral electrophysiological diaschisis could be greatly reversed by early reperfusion and is independent of the lesion size of striatum.