Structure of the dengue virus RNA promoter.

Dengue virus, a single-stranded positive sense RNA virus, is the most prevalent mosquito-borne pathogen in the world. Like all RNA viruses, it uses conserved structural elements within its genome to control essential replicative steps. A 70 nt stem-loop RNA structure (called SLA), found at the 5'-end of the genome of all flaviviruses, functions as the promoter for viral replication. This highly conserved structure interacts with the viral polymerase NS5 to initiate RNA synthesis. Here, we report the NMR structure of a monomeric SLA from dengue virus serotype 1, assembled to high-resolution from independently folded structural elements. The DENV1 SLA has an L-shaped structure, where the top and side helices are coaxially stacked, and the bottom helix is roughly perpendicular to them. Because the sequence is highly conserved among different flavivirus genomes, it is very likely that the three-dimensional fold and local structure of SLA are also conserved among flaviviruses and required for efficient replication. This work provides structural insight into the dengue promoter and provides the foundation for the discovery of new antiviral drugs that target this essential replicative step.

Respiratory Movements at Different Ages.

Background and Objectives: The current study aimed to better understand the changes in respiration that occur with aging in men and women to provide accurate recommendations for breathing exercises to improve health. Materials and Methods: A total of 610 healthy subjects, aged 20 to 59, participated in the study. They performed quiet breathing while wearing two respiration belts (Vernier, Beaverton, OR, USA) at the height of the navel and at the xiphoid process to record abdominal motion (AM) and thoracic motion (TM), respectively. Vital capacity, representing maximal inhalation movement, was measured using a spirometer (Xindonghuateng, Beijing, China). After exclusion, 565 subjects (164 men, aged 41 ± 11; 401 women, aged 42 ± 9) were included for statistical analysis using the Kruskal-Wallis U test and stepwise multiple linear regression. Results: Abdominal motion and its contribution to spontaneous breathing were significantly larger for older men, while the contribution of thoracic motion was smaller for older men. There was no significant difference in thoracic motion between the younger and older men. The differences in women's respiratory movements among various ages were mild and negligible. The contribution of thoracic motion to spontaneous breathing in women was larger than in men for those of older ages (40-59 years), but not for those of younger ages (20-39 years). Additionally, men's and women's vital capacities were less in those of older ages, and the men's were larger than the women's. Conclusions: The findings indicate that men's abdominal contribution to spontaneous breathing increased from 20 to 59 years of age due to increased abdominal motion. Women's respiratory movements did not change much with aging. The maximal inhalation movement became smaller with aging for men and women. Healthcare professionals should focus on improving thoracic mobility when addressing health concerns about aging.

The Effects of Charged Amino Acid Side-Chain Length on Diagonal Cross-Strand Interactions between Carboxylate- and Ammonium-Containing Residues in a ß-Hairpin.

The ß-sheet is one of the common protein secondary structures, and the aberrant aggregation of ß-sheets is implicated in various neurodegenerative diseases. Cross-strand interactions are an important determinant of ß-sheet stability. Accordingly, both diagonal and lateral cross-strand interactions have been studied. Surprisingly, diagonal cross-strand ion-pairing interactions have yet to be investigated. Herein, we present a systematic study on the effects of charged amino acid side-chain length on a diagonal ion-pairing interaction between carboxylate- and ammonium-containing residues in a ß-hairpin. To this end, 2D-NMR was used to investigate the conformation of the peptides. The fraction folded population and the folding free energy were derived from the chemical shift data. The fraction folded population for these peptides with potential diagonal ion pairs was mostly lower compared to the corresponding peptide with a potential lateral ion pair. The diagonal ion-pairing interaction energy was derived using double mutant cycle analysis. The Asp2-Dab9 (Asp: one methylene; Dab: two methylenes) interaction was the most stabilizing (-0.79 ± 0.14 kcal/mol), most likely representing an optimal balance between the entropic penalty to enable the ion-pairing interaction and the number of side-chain conformations that can accommodate the interaction. These results should be useful for designing ß-sheet containing molecular entities for various applications.

Insertion of Pro-Hyp-Gly provides 2 kcal mol-1 stability but attenuates the specific assembly of ABC heterotrimeric collagen triple helices.

Collagen is a major structural component of the extracellular matrix and connective tissue. The key structural feature of collagen is the collagen triple helix, with a Xaa-Yaa-Gly (glycine) repeating pattern. The most frequently occurring triplet is Pro (proline)-Hyp (hydroxyproline)-Gly. The reversible thermal folding and unfolding of a series of heterotrimeric collagen triple helices with varying number of Pro-Hyp-Gly triplets were monitored by circular dichroism spectroscopy to determine the unfolding thermodynamic parameters Tm (midpoint transition temperature), ΔHTm (unfolding enthalpy), and ΔGunfold (unfolding free energy). The Tm and ΔGunfold of the heterotrimeric collagen triple helices increased with increasing number of Pro-Hyp-Gly triplets. The ΔGunfold increased by 2.0 ± 0.2 kcal mol-1 upon inserting one Pro-Hyp-Gly triplet into all three chains. The Tm difference between the most stable ABC combination and the second most stable BCC combination decreased with increasing number of Pro-Hyp-Gly triplets, even though the ΔGunfold difference remained the same. These results should be useful for tuning the stability of collagen triple helical peptides for hydrogel formation, recognition of denatured collagen triple helices as diagnostics and therapeutics, and targeted drug delivery.

Longer charged amino acids favor ß-strand formation in hairpin peptides.

Interactions between charged amino acids significantly influence the structure and function of proteins. The encoded charged amino acids Asp, Glu, Arg, and Lys have different number of hydrophobic methylenes linking the backbone to the charged functionality. It remains to be fully understood how does this difference in the number of methylenes affect protein structure stability. Protein secondary structures are the fundamental three-dimensional building blocks of protein structures. ß-Sheet structures are particularly interesting, because these structures have been associated with a number of protein misfolding diseases. Herein, we report the effect of charged amino acid side chain length at two ß-strand positions individually on the stability of a ß-hairpin. The charged amino acids include side chains with a carboxylate, an ammonium, or a guanidinium group. The experimental peptides, fully folded reference peptides, and fully unfolded reference peptides were synthesized by solid phase peptide synthesis and analyzed by 2D NMR methods including TOCSY, DQF-COSY, and ROESY. Sequence specific assignments were performed for all peptides. The chemical shift data were used to derive the fraction folded population and the folding free energy for the experimental peptides. Results showed that the fraction folded population increased with increasing charged amino acid side chain length. These results should be useful for developing functional peptides that adopt the ß-conformation.

A Small Cyclic ß-Hairpin Peptide Mimics the Rbfox2 RNA Recognition Motif and Binds to the Precursor miRNA 20b.

The RNA recognition motif (RRM), which is the most abundant RNA-binding motif in eukaryotes, is a well-structured domain of about 90 amino acids, yet the ß2ß3 hairpin, corresponding to strands 2 and 3 of the ß-sheet, and the intervening loop make essential interactions with RNA in many RRM complexes. A series of small cyclic peptide mimics of the ß2ß3 hairpin of Rbfox2 protein that recognize the terminal loop of precursor miR-20b have been designed to investigate whether the full RNA-binding protein can be mimicked with a minimal structurally preorganized peptide. Within a small library of seven cyclic peptides, a peptide with low-micromolar affinity for the miR-20b precursor was found. NMR spectroscopy titration data suggest that this peptide specifically targets the apical loop of pre-miR-20b. This work shows that it is possible to mimic RNA-binding proteins with designed stable peptides, which provide a starting point for designing or evolving small peptide mimetics of RRM proteins.

MicroRNA-30b Regulates High Phosphorus Level-Induced Autophagy in Vascular Smooth Muscle Cells by Targeting BECN1.

BACKGROUND/AIMS: Autophagy is an evolutionarily conserved mechanism that affects the survival and functions of vascular smooth muscle cells (VSMCs). We explored the role of microRNAs (miRNAs) in regulating autophagy in VSMCs exposed to high phosphorus (Pi) levels. METHODS: VSMCs were isolated from the thoracic aorta of rats and were cultured primarily. Real-time PCR was used to measure the mRNA expression of indicated genes. Western blotting was performed to detect the protein expression of autophagy-related markers. RESULTS: We found that treatment with high Pi levels (1 and 3 mM) activated LC3II expression and promoted autophagic flux in VSMCs. Conversely, treatment with an autophagy inhibitor decreased LC3II expression. Pi stimulation dysregulated the expression of several miRNAs such as miR-18a, miR-21, miR-23a, miR-30b, and miR-31a. However, miR-30b overexpression decreased Pi-induced expression of autophagy-related marker genes such as BECN1, ATG5, and LC3b, whereas miR-30b downregulation increased Pi-induced expression of these genes. In addition, we found that miR-30b directly targeted BECN1. CONCLUSIONS: These data suggest that miR-30b plays an important role in the regulation of high Pi level-induced autophagy in VSMCs by targeting BECN1.

Comparative Study of the Condensing Effects of Ergosterol and Cholesterol.

Cholesterol, due to its condensing effect, is considered an important regulator of membrane thickness. Other sterols, due to their structural similarities to cholesterol, are often assumed to have a universal effect on membrane properties similar to the condensing effect of cholesterol, albeit possibly to different degrees. We used x-ray diffraction to investigate this assumption. By the combination of lamellar diffraction and grazing-angle scattering, we measured the membrane thickness and the tilt-angle distribution of the lipid's hydrocarbon chains. This method is sensitive to phase separation, which is important for examining the miscibility of sterols and phospholipids. Mixtures of ergosterol or cholesterol with dimyristoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, and dioleoylphosphatidylcholine were systematically studied. We found that mixing ergosterol with phospholipids into a single phase became increasingly difficult with higher sterol concentrations and also with higher concentrations of unsaturated lipid chains. The only condensing effect of ergosterol was found in dimyristoylphosphatidylcholine, although the effect was less than one-third of the effect of cholesterol. Unlike cholesterol, ergosterol could not maintain a fixed electron density profile of the surrounding lipids independent of hydration. In dioleoylphosphatidylcholine and palmitoyloleoylphosphatidylcholine, ergosterol made the membranes thinner, opposite to the effect of cholesterol. In all cases, the tilt-angle variation of the chain diffraction was consistent with the membrane thickness changes measured by lamellar diffraction, i.e., a thickening was always associated with a reduction of chain tilt angles. Our findings do not support the notion that different sterols have a universal behavior that differs only in degree.

High phosphorus level leads to aortic calcification via ß-catenin in chronic kidney disease.

AIMS: Vascular calcification is a risk factor for causing cardiovascular events and has a high prevalence among chronic kidney disease (CKD) patients. However, the molecular mechanism underlying this pathogenic process is still obscure. METHODS: Vascular smooth muscle cells (VSMCs) were induced by a concentration of phosphorus (Pi) of 2.5 mM, and were subjected to cell calcification analyses. The effect of high Pi on the Wnt/ß-catenin pathway was measured using a TOP/FOP-Flash reporter assay. The transcriptional regulation of ß-catenin on PIT1 (a type III sodium-dependent phosphate cotransporter) was confirmed by promoter reporter and chromatin immunoprecipitation assays. The 5/6 nephrectomized rat was used as an in vivo model and was fed a high Pi diet to induce aortic calcification. Serum levels of phosphate, calcium, creatine, and blood urea nitrogen were measured, and abdominal aortic calcification was examined. RESULTS: High Pi induced VSMC calcification, downregulated expression levels of VSMC markers, and upregulated levels of osteogenic markers. High Pi activated the Wnt/ß-catenin pathway and ß-catenin activity. ß-Catenin was involved in the process of high Pi-induced VSMC calcification. Further investigation revealed that ß-catenin transcriptionally regulated Pit1, a necessary player in VSMC osteogenic phenotype change and calcification. The in vivo study showed that ß-catenin was involved in rat abdominal aortic calcification induced by high Pi. When knockdown expression of ß-catenin in the rat model was investigated, we found that aortic calcification was reduced. CONCLUSION: These results suggest that ß-catenin is an important player in high phosphorus level-induced aortic calcification in CKD.

Protective effect of metformin on renal injury of C57BL/6J mouse treated with high fat diet.

This study aimed to investigate the protective effect of metformin on renal injury of C57BL/6J mice treated with a high fat diet. High-fat diet for 12 weeks was used to establish the mice model of metabolism syndrome and the intervention of metformin (75 mg x kg(-1) x d(-1)) for 4 weeks, and plasma biochemical indicator and body weight were used to evaluate the model. Sterol regulatory element-binding protein (SERBP)-1c, TNF-α, NADPH Oxidase (NOX)4 mRNA was determined by real time-PCR. Phospho-AMP-activated protein kinase (P-AMPK)α protein was detected by western blotting. Oil Red O staining, Masson staining and HE staining were for observing renal pathological changes. At the end of 12th week, compared with mice on low fat diet (LFD), body weight (BW), the levels of fasting insulin (FINS), plasma and renal triglyceride (TG) were higher and plasma high density lipoprotein (HDL) and insulin sensitivity index (ISI) were significantly lower, but the levels of fasting blood glycemia (FBG), plasma total cholesterol (TC) and renal TC had no changes; Oil Red O staining revealed renal lipids deposition, Masson staining and HE staining revealed glomerular hypertrophy, matrix increasing, and inflammatory cells infiltration in glomerular; the expression of p-AMPKα protein decreased and the expression of SREBP-1c, TNF-α, NOX4 mRNA increased significantly in mouse treated with high fat diet (HFD). Compared with the HFD group, through metformin interventing, metabolic disorders were significantly improved, renal lipids deposition and other pathological changes were ameliorated, the expression of p-AMPKα protein increased and the expression of SREBP-1c, TNF-α, NOX4 mRNA decreased significantly. Metformin improved metabolic disorders, upregulated activity of renal AMPK, diminished the expression of renal SREBP-1c, TNF-α, NOX4 mRNA, decreased accumulation of renal lipids, and prevened renal injury.

[Association of TNF-1031T/C and clinical efficacy of insulin therapy in newly diagnosed type 2 diabetics].

OBJECTIVE: Tumor necrosis factor (TNF ) locus has been a long-standing type 2 diabetes (T2D) candidate gene. Few studies have been conducted on TNF SNP (single nucleotide polymorphism) as rs1799964 (T-1031C), rs1800630 (A-863C) and rs1799724 (C-857T) in T2D. The purpose of this study is to examine the association of TNF SNP and T2D in a case control study and further explore whether these SNPs influence the clinical efficacy of insulin therapy. METHODS: A total of 109 newly diagnosed type 2 diabetics and 168 healthy individuals were recruited. Three tag SNPs (rs1799964 (T-1031C), rs1800630 (A-863C), rs1799724 (C-857T)) were selected across the TNF locus and polymerase chain reaction (PCR) directed sequencing was performed. The patients received Lispro 25 twice daily to achieve glycemic control and they were followed up for 1 year. Plasma glucose level, lipid profile, homeostatic model assessment for insulin resistance (HOMA-IR) and homeostatic model assessment for beta-cell function (HOMA-ß) were compared among groups with different haplotypes of SNPs. RESULTS: Haplotype of TNF-1031C-863C-857C increased the risk of T2D (OR = 2.7, P < 0.05) . Comparing with homozygote of TNF-1031T-863C-857C diabetics (TCC), those carrying CCC allele had higher fasting serum insulin (16.1(12.0-20.3) mU/L) and HOMA-IR (lnHOMA-IR 1.8 ± 0.4) levels (TCC group: 10.6(8.1-14.3) mU/L and 1.42 ± 0.54 respectively, P < 0.05)). One-year insulin treatment decreased HbA1c effectively in both TCC and CCC groups (P < 0.05). However, higher HOMA-IR was still observed in CCC group than that of TCC after normoglycemia (lnHOMA-IR: 2.5(0.9-3.9) vs 1.1(0.8-1.8) respectively, P < 0.05) . Moreover HOMA-ß showed no significant improvement in CCC group as it was in TCC group by the endpoint of follow-up. CONCLUSIONS: TNF-1031C-863C-857C is a risk haplotype for T2D. CCC carrying patients failed to achieve HOMA-ß improvement. And it might be due to increased endogenous HbOMA-IR level comparing with TCC homozygote.

[Study on leuprorelin acetate in treatment of uterine adenomyosis with infertility].

OBJECTIVE: To study clinical efficacy of leuprorelin acetate in treatment of uterine adenomyosis with infertility. METHODS: From January 1,2011 to March 31,2012, 166 cycles in 166 infertile patients combined with uterine adenomyosis undergoing in vitro fertilization embryo transplant (IVF-ET) with long protocol ovum induction by leuprorelin acetate in centre of medical reproduction, ningbo women and children's hospital were studied retrospectively. In the mean time, 200 cycles in 200 infertile patients with tubal factors were enrolled as control group.The volume of uterus and outcome of IVF-ET were compared and studied between two groups. RESULTS: (1) Volume of uterus:in adenomyosis group, after 2-6 cycles of injecting leuprorelin acetate (3.75 mg/28 days), the mean uterine volume was shrinked from (180 ± 73) cm(3) to (86 ± 67) cm(3) (P < 0.05). (2) Outcome of IVF-ET:the rate of embryo implantation was 39.1% in adenomyosis group and 35.8% in control group. The rate of clinical pregnancy was 54.2% in adenomyosis group and 53.7% in control group. The rate of abortion was 4.7% in adenomyosis group and 4.2% in control group. They all did not show statistical differences (P > 0.05). (3) In adenomyosis group, the rate of fertilization, two pronuclear (2PN) and superior embryo were 67.2%(319/475), 60.8% (289/475) and 52.9% (162/306) in patients with failed pregnancy and 74.2% (423/570), 67.7% (386/570) and 62.1% (256/412) in patients with successful pregnancy after IVF-ET, which reached significant difference (P < 0.05). CONCLUSION: Leuprorelin acetate could improve volume of uterine adenomyosis and outcome of pregnancy in patients undergoing IVF-ET.

Sesamin attenuates PM2.5-induced cardiovascular injury by inhibiting ferroptosis in rats.

Objective: This study aimed to elucidate the pharmacological effects of sesamin (Ses) and its mechanism of action towards PM2.5-induced cardiovascular injuries. Method: Forty Sprague Dawley (SD) rats were randomly divided into five groups: a saline control group; a PM2.5 exposure group; and low-, middle-, and high-dose Ses pretreatment groups. The SD rats were pretreated with different concentrations of Ses for 21 days. Afterward, the rats were exposed to ambient PM2.5 by intratracheal instillation every other day for a total of three times. The levels of inflammatory markers, including tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1ß), and interleukin-6 (IL-6), and indicators related to oxidative responses, such as total superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA), were measured in the blood and heart. The expression of ferroptosis-related proteins in heart tissues was determined via western blot and immunohistochemistry. Results: Ses pretreatment substantially ameliorated cardiovascular injuries in rats as evidenced by the decrease in the pathological score and collagen area. The decreased levels of SOD, GSH, and GSH-Px in the heart and serum were inhibited by Ses. In addition, Ses not only notably increased the activity of antioxidant enzymes but also reduced the levels of MDA, CK, LDH, CK-MB, IL-6, TNF-α, IL-1ß, and IL-6. Furthermore, Ses pretreatment upregulated the expression levels of GPX4, SLC7A11, TFRC, and FPN1 and inhibited the expression levels of FTH1 and FTL. Conclusion: Ses pretreatment could ameliorate PM2.5-induced cardiovascular injuries perhaps by inhibiting ferroptosis. Therefore, Ses pretreatment may be a novel strategy for the prevention and treatment of PM2.5-induced cardiovascular injury.

Ultrasound screening for developmental dysplasia of the hip after 4 weeks increases exam accuracy and decreases follow-up visits.

BACKGROUND: Developmental dysplasia of the hip (DDH) is a preventable and treatable disorder in children. Hip ultrasound is recommended for early detection of affected hips. The timing of the initial hip ultrasound and the frequency of subsequent ultrasounds are controversial topics when considering costs and efficiency. METHODS: Registry data from the Taiwanese Screening and Audit System for Developmental Dysplasia of the Hip were obtained for biometry of hip ultrasounds using the Graf classification and relevant demographic data from 2016. Initial screening results and final case management outcomes were compared to determine screening accuracy and the number of visits needed to determine final outcomes. RESULTS: In total, we screened 1683 newborns in 2016. Of the initial cases screened within 28 days (n = 1168), 86.6% were negative, 10.1% positive, and 3.3% intermediate, while of the cases screened after 28 days (n = 515), 97.3% were negative, 0.8% positive, and 1.9% intermediate. Screening of the newborns' final hip outcomes revealed that 1641 (97.6%) were negative, treatment was administered in 8 cases (0.4%), and 34 (2.0%) cases were lost to follow-up. When comparing screening times, screening after 28 days improved specificity (89%-97%), and later screenings were associated with fewer visits needed to confirm hip outcomes (aOR = 0.19, CI95% = 0.10-0.38, p < 0.001) and improved accuracy (aOR = 13.84, CI95% = 4.23-45.26, p < 0.001). CONCLUSION: This study provides evidence of the benefits of screening for DDH after 28 days, namely: reduced false positives, improved screening accuracy, and a reduced requirement for follow-up visits. Delaying screening can also potentially reduce unnecessary parental anxiety, eliminate unnecessary healthcare burdens, and reduce costs. We recommend performing hip ultrasound screening for newborns after 28 days.

Large-area and thin light guide plates fabricated using UV-based imprinting.

As the demand of larger and thinner flat panel display increasing, conventional methods such as injection molding and hot embossing to fabricate light guide plates (LGPs) become difficult and unsuitable. This study reports a low-cost and high-throughput method to fabricate large-size (320 mm x 240 mm, 15" in diagonal) LGPs by using UV-based imprinting process. With the UV-based imprinting process, a large-size LGP with thickness down to 0.8 mm has been successfully fabricated. The optical property of fabricated LGP has been verified. This study has demonstrated the fabrication of large-size and thin LGPs by using UV-based imprinting process, and the possibility of UV-based imprinting process for fabricating other large thin optical elements.

Fabrication of large area resin microlens arrays using gas-assisted ultraviolet embossing.

Large-area microlens arrays are becoming important components in many applications such as LCD-TV diffusers. This paper reports a uniform pressure, low temperature process for their fabrication. The process integrates gas-assisted embossing and UV-curing embossing. During the process, the 230mm x 203mm PMMA substrate is pressed against the stainless-steel stamper coated with UV-curable resin. Under the gas pressuring and UV irradiating, a large array of microlens can be formed. By using this process, high embossing temperature and high embossing pressure can be avoided. Little residual stress is observed in the embossed PMMA substrate. The uniformity of large-area fabrication and optical properties of fabricated resin microlens array have been verified. This study has successfully shown the potential of this gas-assisted UV embossing process for the replication of large-area microstructures.

Flow mediated dilatation of Taiwanese women with normal singleton pregnancies.

OBJECTIVE: To determine the normal values of flow mediated dilatation (FMD) in Taiwanese women with normal singleton pregnancies for the early detection of preeclampsia. MATERIALS AND METHODS: Data of women with normal singleton pregnancies seen at the Tri-Service General Hospital and Taiji Clinic between January 2014 and December 2015 were collected and analyzed. FMD was measured using high-resolution ultrasonography of the brachial artery for the assessment of endothelial function at the first and second trimester. The relationship between the FMD values and maternal gestational age was analyzed. RESULTS: A total of 122 pregnant women were included in the study. Systole FMD values first and second trimester were 9.05 ± 3.72 and 10.93 ± 3.74, respectively; and the diastole were 9.24 ± 3.64 and 11.18 ± 3.93, respectively. FMD and gestational age were positively correlated (systole, p = 0.0175; diastole, p = 0.0149). CONCLUSION: The normal values of FMD in Taiwanese women with normal singleton pregnancies were established, and data suggests that both systolic and diastolic FMD increase with gestational age. Because of the high failure rate, measurement of FMD may not be suitable as a routine clinical examination.

Antiepileptic drug use among women from the Taiwanese Registry of Epilepsy and Pregnancy: Obstetric complications and fetal malformation outcomes.

To investigate antiepileptic drugs (AEDs) prescription and pregnancy outcomes in pregnancies with epilepsy in Taiwan between 2004 and 2015. We retrospectively reviewed data from the Taiwanese Registry of Epilepsy and Pregnancy (TREP). The TREP registry is a voluntary prospective cohort registry, which tracks pregnant women with epilepsy and AED prescription throughout pregnancy, delivery, and early childhood development. All TREP pregnancies (n = 318) that had completed questionnaires up until delivery or had had an unsuccessful pregnancy were analyzed. Over 94.7% of women had been prescribed AEDs during pregnancy, with 69.0% and 25.7% having received monotherapy, or polytherapy, respectively. Among live births, 12 (3.9%) reported malformation. Cesarean section rate was reported higher than usual (54.5%). In 2004, 73.3% of AEDs prescribed were 1st generation, with 1st generation prescription rates falling to only 8.3% of total prescribed in 2015. AED polytherapy also fell during the study period (40.0% to 20.0%). Cesarean sections were found to be higher for women over 35 years, who had generalized epilepsy, or had experienced an obstetric complication during pregnancy term. Binary logistic regression revealed that Cesarean section was associated with maternal complications (OR = 5.11, CI 95% = 1.11-23.51, p = 0.036), while malformations were associated with obstetric complication (OR = 20.46, CI 95% = 4.80-87.21, p<0.001). Both AED risk types were not associated with complications or malformations. Our sample provides a unique insight into the women with epilepsy with AED use during pregnancy. Cesarean section rate was observed to be higher than usual, but malformation rates remained low. Results indicate a decrease in both 1st generation AEDs and proportion of patients receiving polytherapy over the study period. Obstetric complications were associated with Cesarean section. Fetal malformations were significantly associated with obstetric complications. AED risk factors were not significantly associated with either complications or malformations.

Studying antibiotic-membrane interactions via X-ray diffraction and fluorescence microscopy.

Antibiotic drug resistance is a serious issue for the treatment of bacterial infection. Understanding the resistance to antibiotics is a key issue for developing new drugs. We used penicillin and sulbactam as model antibiotics to study their interaction with model membranes. Cholesterol was used to target the membrane for comparison with the well-known insertion model. Lamellar X-ray diffraction (LXD) was used to determine membrane thickness using successive drug-to-lipid molar ratios. The aspiration method for a single giant unilamellar vesicle (GUV) was used to monitor the kinetic binding process of antibiotic-membrane interactions in an aqueous solution. Both penicillin and sulbactam are found positioned outside the model membrane, while cholesterol inserts perpendicularly into the hydrophobic region of the membrane in aqueous solution. This result provides structural insights for understanding the antibiotic-membrane interaction and the mechanism of antibiotics.