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1.
Spectrochim Acta A Mol Biomol Spectrosc ; 310: 123893, 2024 Apr 05.
Article in English | MEDLINE | ID: mdl-38290284

ABSTRACT

Flavonoids are a kind of natural polyphenols which are closely related to human health, and the identification of flavonoids with similar structures is an important but difficult issue. We herein easily constructed a powerful fluorescent sensor ensemble by using surfactant cetyltrimethylammoniumbromide (CTAB) encapsulating two commercially available fluorescent probes (F1 and F2) with multi-wavelength emission. Fluorescence measurements illustrate the present sensor ensemble exhibits turn-off responses to flavones and flavonols but ratiometric responses to isoflavones, owing to different FRET processes. The heat map and linear discriminant analysis (LDA) results show that this single sensor can effectively distinguish 6 flavonoids belong to three subgroups by collecting the fluorescence variation at four typical wavelengths. Moreover, it can be applied to identify different flavonoids even in biofluids like serum and urine, providing potential practical application.


Subject(s)
Flavones , Isoflavones , Humans , Flavonoids/chemistry , Flavonols , Surface-Active Agents/chemistry , Fluorescent Dyes/chemistry
2.
Reprod Sci ; 31(4): 917-931, 2024 Apr.
Article in English | MEDLINE | ID: mdl-37989803

ABSTRACT

Preterm premature rupture of membranes (pPROM) poses a significant threat to fetal viability and increases the risk for newborn morbidities. The perinatal period of preterm infants affected by pPROM is often characterized by higher rates of mortality and morbidity, with associated risks of cerebral palsy, developmental delays, compromised immune function, respiratory diseases, and sensory impairments. pPROM is believed to result from a variety of causes, including but not limited to microbially induced infections, stretching of fetal membranes, oxidative stress, inflammatory responses, and age-related changes in the fetal-placental interface. Maternal stress, nutritional deficiencies, and medically induced procedures such as fetoscopy are also considered potential contributing factors to pPROM. This comprehensive review explores the potential etiologies leading to pPROM, delves into the intricate molecular mechanisms through which these etiologies cause membrane ruptures, and provides a concise overview of diagnostic and treatment approaches for pPROM. Based on available therapeutic options, this review proposes and explores the possibilities of utilizing a novel composite hydrogel composed of amniotic membrane particles for repairing ruptured fetal membranes, thereby holding promise for its clinical application.


Subject(s)
Fetal Membranes, Premature Rupture , Premature Birth , Pregnancy , Infant, Newborn , Female , Humans , Infant, Premature , Placenta , Fetal Membranes, Premature Rupture/etiology , Fetal Membranes, Premature Rupture/therapy , Fetal Membranes, Premature Rupture/diagnosis , Premature Birth/etiology , Gestational Age
3.
BMC Pregnancy Childbirth ; 23(1): 825, 2023 Nov 30.
Article in English | MEDLINE | ID: mdl-38037011

ABSTRACT

BACKGROUND: Worldwide, frozen embryo transfer (FET) has become a new strategy for the treatment of infertility. The success of FET is closely related to endometrial receptivity. Does uterine artery Doppler during the implantation window predict pregnancy outcome from the first FET? METHODS: A total of 115 retrospectively collected cycles were included in the study, with 64 cycles of clinical pregnancy and 51 cycles of nonclinical pregnancy; There were 99 nonabsent end-diastolic flow (NAEDF) cycles and 16 absent end-diastolic flow (AEDF) cycles. The differences in uterine artery Doppler findings between different pregnancy outcomes were investigated. The clinical pregnancy rate and spontaneous abortion rate in the NAEDF and AEDF groups were compared. The predictive value of uterine artery Doppler during the implantation window in the success rate of pregnancy from the first FET was also investigated. RESULTS: Between the clinical pregnancy group and the nonclinical pregnancy group, there were no significant differences in the mean resistance index (mRI) (Z = -1.065, p = 0.287), mean pulsatility index (mPI) (Z = -0.340, p = 0.734), and mean peak systolic/end-diastolic velocity(mS/D) (Z = -0.953, p = 0.341); there were significant differences in the mean peak systolic velocity (mPSV) (Z = -1.982, p = 0.048) and mean end-diastolic velocity (mEDV) (Z = -2.767, p = 0.006). Between the NAEDF and AEDF groups, there was no significant difference in the clinical pregnancy rate (χ2 = 0.003, p = 0.959), and there was a significant difference in the spontaneous abortion rate (χ2 = 3.465, p = 0.019). Compared with uterine artery Doppler alone, its combination with artificial abortion history, waist-to-hip ratio, LH (Luteinizing hormone) of P (Progesterone) administration day, mPSV and mEDV had a higher predictive value regarding clinical pregnancy from the first FET [ROC-AUC 0.782, 95% CI (0.680-0.883) vs. 0.692, 95% CI (0.587-0.797)]. CONCLUSIONS: Uterine artery Doppler, particularly mPSV and mEDV during the implantation window, was useful for predicting clinical pregnancy, and AEDF was related to spontaneous abortion in the first trimester. Uterine artery Doppler combined with artificial abortion history, waist-to-hip ratio, LH of P administration day, mPSV and mEDV have a higher predictive value than uterine artery Doppler alone regarding the pregnancy from the first FET.


Subject(s)
Abortion, Spontaneous , Female , Pregnancy , Humans , Uterine Artery/diagnostic imaging , Retrospective Studies , Embryo Transfer , Embryo Implantation , Pregnancy Rate
4.
Mol Genet Genomic Med ; 10(9): e2017, 2022 09.
Article in English | MEDLINE | ID: mdl-35941827

ABSTRACT

BACKGROUND: To investigate the interchromosomal effect (ICE) in chromosome translocation carriers. METHODS: Data on preimplantation genetic testing aneuploidy and structural rearrangements (translocation) were retrospectively collected and classified into a reciprocal translocation group, a Robertsonian translocation group and a control group. According to the carrier's gender and age, all cases underwent further subgroup difference analysis of de novo abnormal embryo rates and the number of chromosomes involved in de novo abnormal embryos. RESULTS: Among the 283 couples who participated in this study, 1076 blastocysts from 352 cycles were collected, and 246 de novo abnormal embryos were included. There was a significant difference in the rate of de novo abnormal embryos among the three groups (p < .05) but no significant difference in the number of de novo abnormal chromosomes in the abnormal embryos (p > .05). Gender and age (classified by 35 years old) had no effect on the de novo abnormal embryo ratios among the translocation carriers (p > .05). However, the de novo abnormal ratio increased with age. The embryo constitution reflected no significant difference between the translocation groups (p > .05). CONCLUSION: The ICE was detected for the translocation carriers. The de novo abnormal embryo ratio increased with age. Gender had no effect on the de novo abnormal embryo ratio. Translocation status played a more important role than age and gender.


Subject(s)
Preimplantation Diagnosis , Adult , Aneuploidy , Female , Genetic Testing , High-Throughput Nucleotide Sequencing , Humans , Pregnancy , Retrospective Studies , Translocation, Genetic
5.
Transpl Immunol ; 72: 101582, 2022 06.
Article in English | MEDLINE | ID: mdl-35307537

ABSTRACT

OBJECTIVE: This study aimed to investigate the duration of progesterone (P) therapy on clinical pregnancy rates as measured by the window of implantation (WOI) in the first cycle of frozen embryo transplantation. METHODS: The study compared the pregnancy rates between 345 cleavage stage transfers and 348 blastocyte transfers of frozen embryos with modified natural cycles in patients from July 1, 2020, to November 30, 2020. Four different P durations were analyzed in the cleavage stage embryo transfer group, i.e., two, three, four, and five days. Five different P durations were analyzed in the blastocyst transfer group, i.e., three, four, five, six, and seven days. RESULTS: The baseline demographics and clinical characteristics of the cleavage stage embryos and blastocyst transfer groups were not comparable. The clinical pregnancy rates following the cleavage stage embryo transfer after two, three, four, and five-day P administration were 45.71%, 44.60%, 38.40%, and 30.43%, respectively (the difference among the subgroups was not significant). Following the blastocyst transfer, the clinical pregnancy rates after three, four, five, six, and seven-day P administration were 50.65%, 63.51%, 60.00%, 54.55%, and 61.54%, respectively (the difference among the subgroups was not significant). In contrast, these two transfer groups showed significantly different clinical pregnancy rates following four and five-day P exposure (P < 0.05). CONCLUSION: For cleavage-stage embryo transfer, the most effective WOI was found between days two and five of P administration. The effective WOI for blastocyst transfer was observed between days three and seven of P administrations.


Subject(s)
Embryo Implantation , Embryo Transfer , Female , Humans , Pregnancy , Pregnancy Rate , Retrospective Studies
6.
Ying Yong Sheng Tai Xue Bao ; 32(11): 3905-3912, 2021 Nov 15.
Article in English | MEDLINE | ID: mdl-34898106

ABSTRACT

The upper and middle reaches of the Hunhe River, with forest coverage rate up to 68%, is an important water source protection area in Liaoning Province. Based on the SWAT model and shadow engineering method, we aimed to estimate the function and service value of water retention in the upper and middle Hunhe River basin from 2000 to 2019, to analyze the spatial heterogeneity, and to provide a scientific basis for the ecological compensation and ecological restoration of the region. Our results showed that the function and service value of water retention in the upper and middle reaches of the Hunhe increased first, and then decreased from 2000 to 2019. The mean annual amount and service value of water retention were 2.46 billion m3 and 15.04 billion yuan, respectively. The spatial distribution of water conservation service functions in the study area was uneven. The areas of high water retention service value were mainly distributed in Qingyuan County and Xinbin County, while the low-value areas were located in municipal districts of Shenyang City. The value of water retention services was obviously different among the ecosystems, with water retention capacity per unit area of forests being much higher than that of grassland, farmland, and cities.


Subject(s)
Ecosystem , Rivers , Conservation of Natural Resources , Forests , Water
7.
Cytogenet Genome Res ; 161(3-4): 120-131, 2021.
Article in English | MEDLINE | ID: mdl-33975305

ABSTRACT

In the present study, we retrospectively recruited 340 patients who underwent spontaneous abortions to investigate chromosomal abnormalities of the conception products in the first trimester. We also performed a relevant analysis of clinical factors. Of these patients, 165 had conception products with chromosomal abnormalities, including 135 aneuploidies, 11 triploidies, 10 complex abnormalities, and 9 segmental aneuploidies. The most common abnormal chromosomes were chromosome 16 in the embryo-transfer group and sex chromosomes in the natural-conception group. The most common abnormal chromosomes in all analyzed maternal age groups were sex chromosomes, 16, and 22. The chromosomal abnormality incidence was related to age and number of spontaneous abortions (both p < 0.05), but not to number of pregnancies, deliveries, induced abortions, or methods of conception (all p > 0.05). The rates of abnormality for chromosomes 12, 15, 20, and 22 increased with age, while the rates for chromosomes 6, 7, 13, and X decreased. In all age groups, aneuploidy was by far the most common abnormality; however, the low-incidence distributions of chromosomal abnormalities were entirely different. Overall, chromosomal aneuploidy was the primary cause of pregnancy loss in the first trimester, and low-frequency abnormalities differed across age subgroups. Chromosomal aberrations were found to be related to maternal age and spontaneous abortion, but not all chromosomal abnormalities increased with age.


Subject(s)
Abortion, Spontaneous/genetics , Aneuploidy , Chromosome Aberrations , Chromosome Disorders/genetics , Cytogenetic Analysis/methods , Pregnancy Trimester, First/genetics , Adult , Female , Fertilization/genetics , Humans , Maternal Age , Monosomy , Pregnancy , Retrospective Studies , Triploidy , Trisomy , Young Adult
8.
ACS Appl Mater Interfaces ; 13(16): 18395-18412, 2021 Apr 28.
Article in English | MEDLINE | ID: mdl-33871966

ABSTRACT

Surfactant assemblies have drawn great attention in fabricating fluorescent sensors as they can provide advantages such as easy preparation, low cost, aqueous detection, high fluorescence stability, high sensitivity to external stimuli, etc. We have devoted the past few years to fluorescent cross-reactive sensors and arrays that are advantageous in differentiating similar analytes and analyzing mixed samples. In this Spotlight on Applications, we introduce our recent advances in developing surfactant assembly-based fluorescent sensors and arrays for discrimination applications. Besides using surfactant assemblies encapsulating fluorophores to fabricate multiple-element-based sensor arrays, we particularly proposed to take advantage of modulation effect of dynamic surfactant assemblies on the photophysical properties of encapsulated fluorophores to construct single-system-based discriminative sensors, which have been successfully applied in differentiation of multiple metal ions and various proteins. The applications of surfactant assembly-based sensors for the detection and discrimination of thiols, amino acids, and explosives are also introduced. Finally, the prospects of further efforts for improving surfactant ensemble sensors and their challenges are discussed.

9.
Front Chem ; 8: 572234, 2020.
Article in English | MEDLINE | ID: mdl-33330361

ABSTRACT

Identification of proteins is an important issue both in medical research and in clinical practice as a large number of proteins are closely related to various diseases. Optical sensor arrays with recognition ability have been flourished to apply for distinguishing multiple chemically or structurally similar analytes and analyzing unknown or mixed samples. This review gives an overview of the recent development of array-based discriminative optical biosensors for recognizing proteins and their applications in real samples. Based on the number of sensor elements and the complexity of constructing array-based discriminative systems, these biosensors can be divided into three categories, which include multi-element-based sensor arrays, environment-sensitive sensor arrays and multi-wavelength-based single sensing systems. For each strategy, the construction of sensing platform and detection mechanism are particularly introduced. Meanwhile, the differences and connections between different strategies were discussed. An understanding of these aspects may help to facilitate the development of novel discriminative biosensors and expand their application prospects.

10.
BMC Pregnancy Childbirth ; 20(1): 542, 2020 Sep 17.
Article in English | MEDLINE | ID: mdl-32943000

ABSTRACT

BACKGROUND: Heterotopic pregnancy occurred after frozen embryo transfer with two D3 embryos, and the case had a history of bilateral salpingectomy due to salpingocyesis. An ectopic heterotopic pregnancy was implanted in the left psoas major muscle, which has not been previously reported. CASE PRESENTATION: A 33-year-old woman presented with left back pain after curettage due to foetal arrest in the uterus without vaginal bleeding and spotting, and painkillers relieved the pain initially. When the painkillers ceased to work, the patient returned to the hospital. The ß-human chorionic gonadotropin (ß-hCG) level remained increased compared with the time of curettage, and a diagnosis of retroperitoneal abdominal pregnancy was suggested by ultrasonography and computerized tomography (CT) with the gestational sac implanted in the left psoas major muscle at the left hilum level. Laparotomy was performed to remove the ectopic pregnancy. During the operation, we carefully separated the adipose tissue between the space of the left kidney door and left psoas major muscle, peeled away the gestational sac that was approximately 50 mm × 40 mm with a 25-mm-long foetal bud, and gave a local injection of 10 mg of methotrexate in the psoas major muscle. Fifty days later, ß-hCG decreased to normal levels. CONCLUSION: It is necessary to pay more attention to the main complaints to exclude rare types of ectopic pregnancies of the pelvis and abdomen after embryo transfer.


Subject(s)
Embryo Transfer , Pregnancy, Heterotopic/diagnostic imaging , Adult , Cryopreservation , Female , Humans , Pregnancy , Tomography, X-Ray Computed , Ultrasonography, Prenatal
11.
J Hum Genet ; 65(5): 455-459, 2020 May.
Article in English | MEDLINE | ID: mdl-32051522

ABSTRACT

Ventriculomegaly with cystic kidney disease (VMCKD) is a rare and severe disorder characterized by cerebral ventriculomegaly, greatly elevated maternal serum alpha-fetoprotein (MSAFP) or amniotic fluid alpha-fetoprotein (AFAFP) levels and kidney disease similar to Finnish congenital nephrosis. Recessive mutations in the CRB2 (NM_173689) gene have been shown to cause the syndrome. Here, we described a nonconsanguineous Chinese family with two fetuses affected with VMCKD. A novel compound heterozygous mutation was identified in the CRB2 gene with co-segregation. One mutation [c.1960G>C (p.A654P)] was inherited from the father, while another mutation [c.3078_c.3093delGGCGCGGCCCCGGCCC (p.L1026Lfs*110)] was inherited from the mother. Preimplantation genetic testing for monogenic disease (PGT-M) was performed for the carrier couple with full informed consent and successfully blocked the inheritance of the disease. Our study has important implications on molecular diagnosis and genetic counseling for VMCKD and extends the mutation spectrum in CRB2 gene.


Subject(s)
Carrier Proteins/genetics , Genetic Testing , Hydrocephalus , Kidney Diseases, Cystic , Membrane Proteins/genetics , Mutation , Preimplantation Diagnosis , Adult , Female , Humans , Hydrocephalus/diagnosis , Hydrocephalus/genetics , Kidney Diseases, Cystic/diagnosis , Kidney Diseases, Cystic/genetics , Male , Pregnancy
12.
Langmuir ; 35(2): 326-341, 2019 01 15.
Article in English | MEDLINE | ID: mdl-30063363

ABSTRACT

The heterogeneous structure and dynamic balancing nature of surfactant aggregates make them attractive in developing fluorescent sensors. They can provide a number of advantages, e.g., enhanced fluorescence stability and quantum yield, detection capability in aqueous solutions, and easy operation. Thus, various strategies have been used to construct surfactant aggregate-based fluorescent sensors. Surfactant aggregates play various roles in different strategies and realize multiple sensing behaviors. Many new functions have been discovered for surfactant aggregates in constructing fluorescent sensors. In this feature article, we briefly summarize the development of surfactant aggregate-based fluorescent sensors and their applications in three different types of sensing: selective sensing, multiple analyte sensing, and cross-reactive sensing. For each type of sensing, the design strategies and the roles of surfactant aggregates are particularly introduced. An understanding of these aspects will help to expand the applications of surfactant assemblies in the sensing field.

13.
Analyst ; 143(16): 3775-3788, 2018 Aug 06.
Article in English | MEDLINE | ID: mdl-29974083

ABSTRACT

Discriminative optical sensors with pattern recognition properties and high-throughput ability have been widely developed as they can distinguish multiple chemically similar analytes. Compared to traditional sensor arrays composed of a series of sensor elements, single-system based discriminative sensors using an array of optical changes at different wavelengths to provide input signals have drawn intensive attention recently. On the one hand, they can provide discrimination ability that is lack in using selective sensors; on the other hand, they can simplify the complex data acquisition process accompanied by multiple-element-based sensor arrays and reduce consumption of sensor samples. This tutorial review gives an overview of the development of single-system based discriminative optical sensors. Different strategies for the construction of single-system based discriminative sensors including dynamic combinatorial libraries, cross-reactive conjugated polymers, DNA G-quadruplex ensembles, combinatorial fluorescent molecular sensors, and fluorophore/surfactant aggregate ensembles are particularly introduced.


Subject(s)
Fluorescent Dyes , G-Quadruplexes , Optics and Photonics , Polymers/chemistry
14.
Fertil Steril ; 108(4): 620-627.e4, 2017 10.
Article in English | MEDLINE | ID: mdl-28863935

ABSTRACT

OBJECTIVE: To develop and validate a new strategy to distinguish between balanced/euploid carrier and noncarrier embryos in preimplantation genetic diagnosis (PGD) cycles for reciprocal translocations and to successfully achieve a live birth after selective transfer of a noncarrier embryo. DESIGN: Retrospective and prospective study. SETTING: In vitro fertilization (IVF) units. PATIENT(S): Eleven patients undergoing mate pair sequencing for identification of translocation breakpoints, followed by clinical PGD cycles. INTERVENTION(S): Embryo biopsy with 24-chromosome testing to determine carrier status of balanced/euploid embryos. MAIN OUTCOME MEASURE(S): Definition of translocation breakpoints and polymerase chain reaction (PCR) diagnostic primers, correct diagnosis of euploid embryos for carrier status, and a live birth with a normal karyotype after transfer of a noncarrier embryo. RESULT(S): In 9 of 11 patients (82%), translocation breakpoints were successfully identified. In four patients with a term PGD pregnancy established with a balanced/euploid embryo of unknown carrier status, the correct carrier status was retrospectively determined, matching with the cytogenetic karyotype of the resulting newborns. In a prospective PGD cycle undertaken by a patient with a 46,XY,t(7;14)(q22;q24.3) translocation, the four balanced/euploid embryos identified comprised three carriers and one noncarrier. Transfer of the noncarrier embryo resulted in birth of a healthy girl who was subsequently confirmed with a normal 46,XX karyotype. CONCLUSION(S): The combination of mate pair sequencing and PCR breakpoint analysis of balanced reciprocal translocation derivatives is a novel, reliable, and accurate strategy for distinguishing between carrier and noncarrier balanced/euploid embryos. The method has potential application in clinical PGD cycles for patients with reciprocal translocations or other structural rearrangements.


Subject(s)
Embryo Transfer/methods , Fertilization in Vitro , Genetic Carrier Screening/methods , Preimplantation Diagnosis/methods , Translocation, Genetic , Adult , Female , Fertilization in Vitro/methods , Humans , Infant, Newborn , Karyotyping , Male , Ploidies , Polymerase Chain Reaction/methods , Pregnancy , Prenatal Diagnosis/methods , Retrospective Studies
15.
Prenat Diagn ; 36(2): 154-62, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26676536

ABSTRACT

OBJECTIVE: Currently, our understanding of the nature and reproductive potential of blastocysts associated with trophectoderm (TE) lineage chromosomal mosaicism is limited. The objective of this study was to first validate copy number variation sequencing (CNV-Seq) for measuring the level of mosaicism and second, examine the nature and level of mosaicism in TE biopsies of patient's blastocysts. METHOD: TE biopy samples were analysed by array comparative genomic hybridization (CGH) and CNV-Seq to discriminate between euploid, aneuploid and mosaic blastocysts. RESULTS: Using artificial models of TE mosaicism for five different chromosomes, CNV-Seq accurately and reproducibly quantitated mosaicism at levels of 50% and 20%. In a comparative 24-chromosome study of 49 blastocysts by array CGH and CNV-Seq, 43 blastocysts (87.8%) had a concordant diagnosis and 6 blastocysts (12.2%) were discordant. The discordance was attributed to low to medium levels of chromosomal mosaicism (30-70%) not detected by array CGH. In an expanded study of 399 blastocysts using CNV-Seq as the sole diagnostic method, the proportion of diploid-aneuploid mosaics (34, 8.5%) was significantly higher than aneuploid mosaics (18, 4.5%) (p < 0.02). CONCLUSION: Mosaicism is a significant chromosomal abnormality associated with the TE lineage of human blastocysts that can be reliably and accurately detected by CNV-Seq.


Subject(s)
Blastocyst/metabolism , DNA Copy Number Variations/genetics , Down Syndrome/diagnosis , Mosaicism , Preimplantation Diagnosis/methods , Turner Syndrome/diagnosis , Comparative Genomic Hybridization , Cryopreservation , Down Syndrome/genetics , Embryo Transfer , Female , Humans , Pregnancy , Sequence Analysis, DNA/methods , Trophoblasts/metabolism , Turner Syndrome/genetics
16.
ACS Appl Mater Interfaces ; 7(40): 22487-96, 2015 Oct 14.
Article in English | MEDLINE | ID: mdl-26414441

ABSTRACT

A particular bispyrene fluorophore (1) with two pyrene moieties covalently linked via a hydrophilic spacer was synthesized. Fluorescence measurements reveal that the fluorescence emission of 1 could be well modulated by a cationic surfactant, dodecyltrimethylammonium bromide (DTAB). Protein sensing studies illustrate that the selected ensemble based on 1/DTAB assemblies exhibits ratiometric responses to nonmetalloproteins and turn-off responses to metalloproteins, which can be used to differentiate the two types of proteins. Moreover, negatively charged nonmetalloproteins can be discriminated from the positively charged ones according to the difference in ratiometric responses. Fluorescence sensing studies with control bispyrenes indicate that the polarity of the spacer connecting two pyrene moieties plays an important role in locating bispyrene fluorophore in DTAB assemblies, which further influences its sensing behaviors to noncovalent interacting proteins. This study sheds light on the influence of the probe structure on the sensing performance of a fluorescent ensemble based on probe and surfactant assemblies.


Subject(s)
Metalloproteins/chemistry , Pyrenes/chemistry , Surface-Active Agents/chemistry , Dynamic Light Scattering , Fluorescent Dyes/chemistry , Ions/chemistry , Metalloproteins/metabolism , Metals/chemistry , Quaternary Ammonium Compounds , Spectrometry, Fluorescence , Water/chemistry
17.
Reprod Biomed Online ; 31(1): 62-70, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25985995

ABSTRACT

Next-generation sequencing is emerging as a reliable and accurate technology for pre-implantation genetic diagnosis (PGD) of aneuploidies and translocations. The aim of this study was to extend the clinical utility of copy number variation sequencing (CNV-Seq) to the detection of small pathogenic copy number variations (CNVs) associated with chromosome disease syndromes. In preliminary validation studies, CNV-Seq was highly sensitive and specific for detecting small CNV in whole-genome amplification products from three replicates of one and five cell samples, with a resolution in the order of 1-2 Mb. Importantly, the chromosome positions of all CNV were correctly mapped with copy numbers similar to those measured in matching genomic DNA samples. In seven clinical PGD cycles where results were obtained for 34 of 35 blastocysts, CNV-Seq identified 18 blastocysts with aneuploidies, one with an aneuploidy and a 4.98 Mb 5q35.2-qter deletion associated with Sotos syndrome, one with a 6.66 Mb 7p22.1-pter deletion associated with 7p terminal deletion syndrome and 14 with no detectable abnormalities that were suitable for transfer. On the basis of these findings, CNV-Seq displays the hallmarks of a comprehensive PGD technology for detection of aneuploidies and CNVs that are known to affect the development and health of patient's embryos.


Subject(s)
Chromosome Disorders/diagnosis , Embryo, Mammalian/physiology , High-Throughput Nucleotide Sequencing , Preimplantation Diagnosis/methods , Aneuploidy , DNA Copy Number Variations , Genome, Human , Humans , Sensitivity and Specificity
18.
Neuron ; 85(5): 1086-102, 2015 Mar 04.
Article in English | MEDLINE | ID: mdl-25741729

ABSTRACT

Circadian behavior in mammals is orchestrated by neurons within the suprachiasmatic nucleus (SCN), yet the neuronal population necessary for the generation of timekeeping remains unknown. We show that a subset of SCN neurons expressing the neuropeptide neuromedin S (NMS) plays an essential role in the generation of daily rhythms in behavior. We demonstrate that lengthening period within Nms neurons is sufficient to lengthen period of the SCN and behavioral circadian rhythms. Conversely, mice without a functional molecular clock within Nms neurons lack synchronous molecular oscillations and coherent behavioral daily rhythms. Interestingly, we found that mice lacking Nms and its closely related paralog, Nmu, do not lose in vivo circadian rhythms. However, blocking vesicular transmission from Nms neurons with intact cell-autonomous clocks disrupts the timing mechanisms of the SCN, revealing that Nms neurons define a subpopulation of pacemakers that control SCN network synchrony and in vivo circadian rhythms through intercellular synaptic transmission.


Subject(s)
Circadian Clocks/physiology , Circadian Rhythm/physiology , Neurons/physiology , Neuropeptides/biosynthesis , Suprachiasmatic Nucleus/physiology , Animals , Biological Clocks/physiology , Mice , Mice, Inbred C57BL , Mice, Transgenic
19.
J Neurosci ; 35(5): 1905-20, 2015 Feb 04.
Article in English | MEDLINE | ID: mdl-25653351

ABSTRACT

GABAergic synaptic transmission plays an important role in resetting and synchronizing circadian rhythms in the suprachiasmatic nucleus (SCN). Although the circadian modulation of intrinsic membrane currents and biochemical signaling have been examined in the SCN, the modulation of specific synaptic pathways within the SCN is unexplored. In addition, little is known about the functional properties of these pathways, including which ones involve GABAA receptors (GABAA-Rs). In brain slices obtained from mice, we examined synaptic responses originating from the SCN neurons expressing vasoactive intestinal peptide (VIP+ neurons). Focusing on the local projection within the ventromedial SCN, we found that VIP+ afferents provided input onto 49% of neurons with a preference for VIP-negative (VIP-) neurons. Responses were mediated by GABAA-Rs. The projection was sparsely connected and preferentially targeted a subset of SCN neurons unrelated to postsynaptic VIP expression. For most aspects of VIP+ network output, there was no circadian regulation. Excitability and spontaneous firing of the presynaptic VIP+ neurons were unchanged between day and night, and their network connectivity and synaptic function up through the evoked synaptic conductance were also unchanged. On the other hand, VIP+ input onto VIP- neurons became less inhibitory at night suggesting a postsynaptic alteration in the coupling of GABAA-R conductances to action potential firing. These data suggest that components of the VIP network and its synaptic output up through GABAA-R opening are invariant during the circadian cycle, but the effect on action potential firing is modulated by postsynaptic processes occurring after GABAA-R channel opening.


Subject(s)
Circadian Rhythm , GABAergic Neurons/metabolism , Neurons, Afferent/metabolism , Receptors, GABA-A/metabolism , Suprachiasmatic Nucleus/metabolism , Synaptic Potentials , Vasoactive Intestinal Peptide/metabolism , Action Potentials , Animals , Female , GABAergic Neurons/physiology , Male , Mice , Neurons, Afferent/physiology , Suprachiasmatic Nucleus/cytology , Suprachiasmatic Nucleus/physiology , Synapses/metabolism , Synapses/physiology , Vasoactive Intestinal Peptide/genetics , gamma-Aminobutyric Acid/metabolism
20.
ACS Appl Mater Interfaces ; 6(18): 16156-65, 2014 Sep 24.
Article in English | MEDLINE | ID: mdl-25188720

ABSTRACT

Lanthanides are valuable nonrenewable resources and widely used in a variety of industries. Detection and identification of lanthanide ions are in high demand but challenging because of the similarity among lanthanide ions. In the present work, a fluorescent sensor array of three cationic bispyrene derivatives mixed with anionic surfactant assemblies was developed. The sensor array exhibits cross-reactive responses to lanthanide ions when tested in aqueous solution. The combination of fluorescence variations at both monomer and excimer emission of each of the bispyrene sensor elements provides a six-signal recognition pattern for lanthanide ions. Principle component analysis illustrates that the sensor array could at least identify 6 of the 14 similar lanthanide ions including La(3+), Pr(3+), Nd(3+), Eu(3+), Ho(3+), and Er(3+). UV-vis absorption measurements rule out the possibility of binding lanthanides with fluorophores. Fluorescence titration experiments in both cationic and neutral surfactant aqueous solutions reveal that the three fluorophores show slight fluorescence responses to the lanthanide ions, indicating that electrostatic attraction between lanthanide ions and anionic surfactant plays an important role in the sensing behavior of the sensor array. Control experiments with divalent metal ions find no cross-reactive responses, suggesting that the stronger electrostatic interaction with trivalent lanthanide ions is responsible for the multiple fluorescence responses.

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