bluuebunny/crossref_metadata_2025 huggingface dataset:
29 Time-lapse analysis of bovine embryos derived after invitro fertilization from vitrified and fresh oocytes
Authors: Velez D. A. , Atashi H. , Dewulf J. , Smits K. , Van Soom A.
Published: December 2019

Oocyte vitrification enables the long-term conservation of female genetic resources. However, obtaining viable bovine embryos from vitrified oocytes has proven to be difficult. Embryo development is a dynamic process, and critical stages can go unnoticed with the use of traditional morphologic assessments. Therefore, the aim was to evaluate morphokinetic parameters in embryos derived from fresh versus vitrified oocytes. After 22h of IVM, oocytes were divided into (1) oocytes surrounded by multiple layers of cumulus cells (COCs; n=275) and (2) oocytes partially denuded, leaving only corona radiata (CR; n=178). Then, two-thirds of the CR oocytes were subjected to one of two vitrification protocols as follows: high concentration of cryoprotectants (CR-H; n=171; Ortiz-Escribano et al. 2016 Theriogenology 86, 635-641) and low concentration of cryoprotectants (CR-L; n=163; Ishii et al. 2018 J. Reprod. Develop. 6). After warming, vitrified oocytes were incubated ~2h in maturation medium. Invitro fertilization and culture were performed simultaneously for all groups. Four to eight zygotes from each group were assigned randomly to time lapse (Primo Vision; Vitrolife), and group culture was performed in leftovers as a control. Differences between groups in survival (intact zygotes after IVF), cleavage, and blastocyst rates were evaluated by logistic regression. Morphokinetic data (time to reach first (1-2 cells), second (3-4 cells), third (5-8), fourth (9-16 cells), fifth (>16 cells), cleavage, and blastocyst stage, and time in lag phase) were investigated. Survival rates in COCs (98%) and CR (96%) groups were not different; CR-H (87%) showed lower survival than COCs but similar survival to that of CR. Group CR-L (82%) had a lower survival rate than the rest of the groups (P<0.05). Higher cleavage rate (83%) was found in COCs compared with the rest of the groups. The CR and CR-H groups showed similar cleavage rate (65 and 55%, respectively), whereas CR-L had a lower cleavage rate (42%) than other groups (P<0.05). As expected, both vitrified groups showed lower blastocyst rates (4% for CR-H and 10% for CR-L) than fresh COCs (44%; P<0.05). Morphokinetics measures were affected by the treatments: time to reach first cleavage was similar for COCs (35.3h), CR (38.4h), and CR-L (34.8h), whereas CR-H was slower (42.4h). However, the fourth division was reached earlier by CR-H (51.7h), which was significantly faster than for CR-L (88.8 h; P<0.05) but similar for COCs (71.3h) and CR (60.7h). In conclusion, more morphokinetic data are needed to compare different vitrification methods and confirm whether time-lapse analysis can be used to predict blastocyst outcome after oocyte vitrification.
57 Primordial germ cell distribution in the horse fetal gonad
Authors: Scarlet D. , Reichart U. , Podico G. , Ellerbrock R. , Canisso I. , Walter I. , Aurich C.
Published: December 2019

Germ cell development and differentiation is a complex process associated with down-regulation of stem cell-associated genes and up-regulation of markers of germ cell differentiation and meiosis. In horses, putative primordial germ cells (PGCs) were identified outside the gonads starting 20 days after ovulation (Curran et al. 1997 Equine Vet. J. Suppl. 25, 72-76). However, no information is available after the time when these cells enter the gonad. The aim of this study was to identify, localise, and quantify PGCs in fetal male and female gonads. Twelve (5 males and 7 females) equine fetuses were collected transcervically 60 days after ovulation. Fetuses were entirely fixed in formaldehyde and gonads were subsequently localised. Fetal gonads underwent multiplex immunofluorescence staining for pre-meiotic germ cell marker LIN28, stem-cell associated marker CD117, and cell proliferation marker Ki67. Specificity of the primary antibodies for equine samples has been first validated. Quantification of fluorescence positive areas for LIN28, CD117, and Ki67 in the fetal gonads was done using a macro for ImageJ. Furthermore, we analysed the co-localization of above-mentioned markers using the same macro. Statistical comparison for differences between males and females was performed using non-parametric tests. In both sexes, PGCs proliferated as determined by double immunofluorescence of Ki67 and LIN28. Protein expression of LIN28 and Ki67 was highly correlated (r=0.92; P=0.003). In the fetal female gonad, PGCs were organised in cord-like structures localised in the cortical region, but there were also LIN28+ cells in the surface germinal epithelium. In the fetal male gonad, PGCs were restricted to the already developed tubular structures. Fewer LIN28+ cells (3.0±0.4% vs. 4.5±0.3%; P<0.05) were present in female than in male gonads. The distribution pattern of the stem cell factor receptor CD117 was similar to LIN28, as 86.8±3.2% of LIN28+ cells in females and 84.6±4.7% in males were also CD117+. However, immunofluorescent co-localization analysis revealed a subpopulation of CD117+ cells (43.1±8.1% in females and 46.1±6.1% in males), which did not show an overlap with LIN28. These were presumably stem cells localised in the medullar area of the gonad. In summary, we analysed for the first time spatial distribution of PGCs in fetal equine gonads. We demonstrated LIN28 to be a specific marker for PGCs also in the horse gonad, which is in agreement with the situation in human and other species. Furthermore, we identified stem cells and described their localization in the fetal equine gonad. Nevertheless, the temporal distribution of PGC and stem cells in the developing horse gonad and the role of LIN28 in the maintenance of the germline stem cell state still need to be investigated.
59 Genome activation in intracytoplasmic sperm injection-derived horse embryos
Authors: Goszczynski D. E. , Tinetti P. , Choi Y. H. , Hinrichs K. , Ross P. J.
Published: December 2019

During pre-implantation development, embryos go through a critical period of embryonic genome activation (EGA). The timing of EGA is species specific, but little is known in horse embryos. Here, we aimed to characterise EGA in equine embryos produced by intracytoplasmic sperm injection. Embryos were produced by intracytoplasmic sperm injection of oocytes from 3 mares. Two embryos from each mare at each of 8 developmental stages (MII, zygote, 2-cell, 4-cell, 8-cell, 16-cell, morula, and blastocyst) were individually analysed by RNA-seq. Differential expression was evaluated using binomial Wald tests with an absolute logFC (fold change) threshold of 1 in the DESEqn 2R package. We found that EGA occurred in a two-step fashion. Minor EGA took place during the 2-cell to 4-cell transition, and featured up-regulation of 751 genes and discrete down-regulation of 60 genes in 4-cell embryos compared with 2-cell embryos. Differentially upregulated genes were enriched in gene ontology terms related to transcriptional activator activity, homeobox domains, and nucleosome assembly. Major EGA occurred during the 4-cell to 8-cell transition and included the largest number of differentially expressed genes (n=2,023) between consecutive stages. This period also featured the first massive transcript downregulation (n=816). Upregulated genes were enriched in gene ontology terms related to ribosomal assembly, translation, and RNA modification. Additionally, we observed that the number of intronic sequences was significantly higher from the 4-cell stage onward, indicating active transcription in comparison to oocytes, zygotes, and 2-cell embryos. To evaluate the timing of paternal genome activation, we used whole-genome sequencing data from the parents (average genome coverage of 19×) to quantify allele-specific expression. The average number of informative SNPs in exons, i.e. SNPs with alternative homozygous genotypes from the sire (AA mare - BB sire), was 26 128 per mare, corresponding to 7696 genes. Parental-specific transcript abundance was determined for each embryo, with an average of 1,911±865 informative SNPs detected per sample. Paternal alleles were considered expressed when they reached 10% of the maternal count. Across development, paternal transcripts became appreciable at the 4-cell stage, with 14.15±7.60% of the informative SNPs exhibiting paternal expression, and increased thereafter until reaching a maximum of 96.34% at the blastocyst stage. Overall, this work demonstrates that EGA in horse embryos starts at the 4-cell stage and achieves its main activation at the 8-cell stage. Further analysis will be performed to detail paternal vs. maternal gene expression at the different embryonic stages.
69 Phosphorylation of mechanistic target of rapamycin in porcine blastocyst-stage embryos is dependent on the concentration of glutamine in the medium
Authors: Chen P. , Spate L. , Prather R.
Published: December 2019

Supplementation of glutamine to porcine embryo culture medium improves blastocyst development, increases leucine consumption, and enhances mitochondrial activity. In cancer cells, glutamine has been implicated in the phosphorylation and activation of mechanistic target of rapamycin (MTOR) to support rapid cellular proliferation. The objective of this study was to determine if phosphorylation of MTOR in porcine blastocysts was dependent upon the concentration of glutamine in the medium and presence of leucine, another activator of MTOR. Presumptive zygotes (n=732 per treatment across 6 replicates) were split into four groups and cultured in 0, 1, 3.75, or 10mM GlutaMAX, an L-glutamine alternative, with or without 0.2mM leucine. On Day 6, percentages of embryos developed to the blastocyst stage were recorded. Blastocyst-stage embryos (n=100 per treatment) were collected for immunoblotting to detect total MTOR and phosphorylated (Ser2448) MTOR (pMTOR) with 3 replicates per target. To assess MTOR and lysosomal colocalization as another indicator of MTOR activation, blastocyst-stage embryos (n=15 per treatment) were fixed, probed with antibodies against MTOR and lysosomal associated membrane protein 1 (LAMP1), and imaged on a confocal microscope. Data were analysed by using the GLM procedure in SAS 9.4 (SAS Institute Inc.), and differences between means were detected by using one-way ANOVA followed by a least significant difference test with P<0.05 declared significant. Compared with the other treatments, culture in 0mM GlutaMAX resulted in decreased blastocyst development with (26.2±4.8%) or without (23.2±3.0%) leucine. Embryos cultured in 3.75mM GlutaMAX had increased development to the blastocyst stage (+Leu: 49.7±4.2%; −Leu: 46.2±2.6%) compared with culture in 1mM (+Leu: 39.5±3.5%; −Leu: 37.8±2.1%). In the presence of leucine, culture in 0mM GlutaMAX resulted in decreased pMTOR abundance compared with 3.75 or 10mM GlutaMAX. In the absence of leucine, embryos cultured in 0mM GlutaMAX had decreased abundance of total MTOR compared with all other groups, and pMTOR abundance was decreased in embryos cultured in 0 or 1mM compared with 3.75 or 10mM GlutaMAX. Furthermore, embryos cultured in 0mM GlutaMAX had decreased colocalization of MTOR and LAMP1 compared with those cultured in 3.75 or 10mM GlutaMAX. Therefore, glutamine supplementation is sufficient to prompt MTOR phosphorylation in porcine embryos. Further analyses are examining the phosphorylation status of MTOR downstream targets and effects of inhibiting enzymes involved in glutaminolysis.
Funding was provided by the United States Department of Agriculture, National Institute of Food and Agriculture (2019-67011-29543) and Food for the 21st Century at the University of Missouri.
75 Expression of lipid metabolism-related genes in bovine embryos cultured invitro with diacylglycerol acyltransferase-1 inhibitor
Authors: Cañón-Beltrán K. , Giraldo-Giraldo J. , Cajas Y. N. , Vásquez N. , Leal C. L. V. , Gutiérrez-Adán A. , González M. E. , Rizos D.
Published: December 2019

Intracellular lipids accumulated in embryos produced invitro have been linked to reductions in both quality and post-cryopreservation viability. Diacylglycerol O-acyltransferase 1 (DGAT1) is an enzyme that catalyzes the final step in triglyceride synthesis, which is a major component of the lipid droplets in embryos. Thus, the aim of this study was to evaluate the development and quality, in regard to changes in the expression of candidate genes of lipid metabolism, of bovine blastocysts cultured invitro with DGAT1 inhibitor (A922500 Sigma-Aldrich). Zygotes were cultured in synthetic oviduct fluid supplemented with 5% FCS (TC) or in TC supplemented with 10 or 50 µM DGAT1 inhibitor (T10 and T50, respectively) or TC with 0.1% dimethyl sulfoxide (TDMSO: vehicle for DGAT1 dilution) from 54h post-insemination (hpi; major embryonic genome activation, EGA) until Day 8. Cleavage rate (48 hpi) and blastocyst yield (Day 7-8) were evaluated. Day 7 blastocysts were snap-frozen in LN2 for gene expression analysis. The mRNA abundance of candidate genes related to lipid metabolism (DGAT1, PLIN2, GLUT5, GLUT1, GPX1, PPAR1b, and G6PD) was measured by quantitative PCR. The H2AFZ and ACTB genes were used as housekeeping genes. Statistical analysis was assessed by one-way analysis of variance. No differences were found in cleavage rate, whereas blastocyst yield at Day 8 was higher (P<0.05) for T50 (30.5±0.5%) and TC (29.7±0.5%) compared with T10 (25.6±0.5%) and TDMSO (27.1±0.8%). The expression of genes regulating lipid droplet formation (DGAT1 and PLIN2) was down-regulated in embryos of T10 and T50 groups compared with controls (P<0.05) only for DGAT1, whereas no differences were observed for PLIN2. The expression of GLUT5, a fructose metabolism transporter gene, was only increased in embryos in the T10 group, whereas the relative abundance of GLUT1 (involved in glucose metabolism) was up-regulated in T10 and T50 groups compared with controls (P<0.05). The GPX1 gene was decreased significantly in embryos from both DGAT1-inhibitor groups compared with the controls. The expression profile of PPAR1b and G6PD, lipid metabolism-regulating genes, were not different among embryo groups. In conclusion, supplementation of embryo culture with DGAT1 inhibitor improves development and blastocyst quality in terms of the expression of lipid metabolism-regulating genes, supporting a relationship between lipid metabolism and embryo feature.
Funding was provided by MINECO-Spain AGL2015-70140-R & RTI2018-093548-B-I00; J. Giraldo-Giraldo COLCIENCIAS 727/2015-Colombia; Y. N. Cajas, SENESCYT-Ecuador; C. L. V. Leal, FAPESP-Brasil 2017/20339-3.
94 Efficient generation of myostatin mutation in Korean beef cattle
Authors: Gim G. , Jin Y. , Yum S. , Park H. , Lee J. , Koo S. , Lee W. , Goo J.
Published: December 2019

In recent years, CRISPR/Cas9 has become known as a powerful tool for generating genetically modified cells or animal models with its site-specific gene editing capability. Because myostatin (MSTN) is associated with muscle differentiation and growth, ablation of MSTN would be ideal for studying phenotypes associated with muscle growth in beef cattle. The aim of this study is to produce Korean beef cattle with MSTN mutation through microinjection of CRISPR/Cas9. For this study, ovaries of Korean beef cattle were obtained from slaughterhouses; 122 immature oocytes were aspirated from 8 Korean beef cattle. The oocytes were matured in tissue culture medium 199-based medium for 22h and inseminated with frozen-thawed semen to produce zygotes. All 122 zygotes were directly injected with 200ng of Cas9 mRNA and 100ng of sgRNA for MSTN after IVF and were cultured for 7 days in two-step chemically defined medium. Each embryo from the individual cattle was cultured separately to distinguish its lineage. After 7 d, 29 (23.8±8.9) blastocysts were recovered, and all blastocysts were assessed with T7 endonuclease (T7E1) mutation assay to check for mutations at the MSTN locus. Based on T7E1, 26 out of 29 blastocysts (89.7±10.5%) had heterozygous mutation in the MSTN gene. After analysis, additional blastocysts were produced and transferred into 14 surrogates. All surrogates were diagnosed for pregnancy via rectal palpation and ultrasonography at Day 55, at which time normal fetuses and embryonic sacs were confirmed in 10 surrogates. In conclusion, these data demonstrate that CRISPR/Cas9 could be employed as an efficient method to genetically modify the MSTN gene in Korean beef cattle.
95 Obtaining birds with chimeric gonads using invitro lentiviral transduction of primordial germ cells
Authors: Glumakova K. A. , Mityaeva O. N. , Antonova E. N. , Glazova O. V. , Komarchev A. S. , Volchkov P. Y.
Published: December 2019

Avian primordial germ cells (PGCs) have unique migration capacity towards the gonads via the bloodstream. Therefore, PGCs invitro genome modification is a dominant approach for poultry genetic modification. The aim of this study was to improve cultivation conditions of PGCs in terms of proliferation activity and further analyse their migration abilities. We isolated PGCs from whole blood cells collected from the embryonic dorsal aorta (Hamburger-Hamilton (HH) stage 14-17) and cultured them without feeder cells in customized avian knockout Dulbecco's modified Eagle's medium basal medium supplemented with fibroblast growth factor 2, Activin A and B-27 supplement containing insulin. We analysed the number of cells during the month of culture. Proliferation of PGCs increased in the first 7d of culture when both BMP4 and Activin A growth factors were added to the medium (four out of seven samples). Kinetic of stemness and PGC-marker genes (Nanog, PouV, DAZL) were similar to expression in mature gonadal PGCs (gPGCs). Expression levels of marker genes were significantly greater in the freshly isolated PGCs compared to the gPGCs and decreased during cultivation. In order to confirm the migration activity of the cultivated PGCs, cells were labelled with lentivirus (ZsGreen) and injected into the embryo blood stream (HH 14-17). Gonads from the recipient embryos were retrieved (HH 20-26) and analysed. Due to the low level of fluorescence in mature cells, the presence of transgenic PGC in the gonads was detected by ZsGreen-specific quantitative PCR. We confirmed that the cultured cells maintained migration activity, and efficient engraftment was detected in 75% of embryos.
MiR155‐5p in adventitial fibroblasts‐derived extracellular vesicles inhibits vascular smooth muscle cell proliferation via suppressing angiotensin‐converting enzyme expression
Authors: Ren Xing‐Sheng , Tong Ying , Qiu Yun , Ye Chao , Wu Nan , Xiong Xiao‐Qing , Wang Jue‐Jin , Han Ying , Zhou Ye‐Bo , Zhang Feng , Sun Hai‐Jian , Gao Xing‐Ya , Chen Qi , Li Yue‐Hua , Kang Yu‐Ming , Zhu Guo‐Qing
Published: December 2019

Proliferation of vascular smooth muscle cells (VSMCs) plays crucial roles in vascular remodelling and stiffening in hypertension. Vascular adventitial fibroblasts are a key regulator of vascular wall function and structure. This study is designed to investigate the roles of adventitial fibroblasts‐derived extracellular vesicles (EVs) in VSMC proliferation and vascular remodelling in normotensive Wistar‐Kyoto rat (WKY) and spontaneously hypertensive rat (SHR), an animal model of human essential hypertension. EVs were isolated from aortic adventitial fibroblasts of WKY (WKY‐EVs) and SHR (SHR‐EVs). Compared with WKY‐EVs, miR155‐5p content was reduced, while angiotensin‐converting enzyme (ACE) content was increased in SHR‐EVs. WKY‐EVs inhibited VSMC proliferation of SHR, which was prevented by miR155‐5p inhibitor. SHR‐EVs promoted VSMC proliferation of both strains, which was enhanced by miR155‐5p inhibitor, but abolished by captopril or losartan. Dual luciferase reporter assay showed that ACE was a target gene of miR155‐5p. MiR155‐5p mimic or overexpression inhibited VSMC proliferation and ACE upregulation of SHR. WKY‐EVs reduced ACE mRNA and protein expressions while SHR‐EVs only increased ACE protein level in VSMCs of both strains. However, the SHR‐EVs‐derived from the ACE knockdown‐treated adventitial fibroblasts lost the roles in promoting VSMC proliferation and ACE upregulation. Systemic miR155‐5p overexpression reduced vascular ACE, angiotensin II and proliferating cell nuclear antigen levels, and attenuated hypertension and vascular remodelling in SHR. Repetitive intravenous injection of SHR‐EVs increased blood pressure and vascular ACE contents, and promoted vascular remodelling in both strains, while WKY‐EVs reduced vascular ACE contents and attenuated hypertension and vascular remodelling in SHR. We concluded that WKY‐EVs‐mediated miR155‐5p transfer attenuates VSMC proliferation and vascular remodelling in SHR via suppressing ACE expression, while SHR‐EVs‐mediated ACE transfer promotes VSMC proliferation and vascular remodelling.
The political constraints for civil service reform in Georgia: history, current affairs, prospects and challenges
Authors: Kvashilava Bakur
Published: December 2019

This study provides an examination (and an evaluation) of the reforms of the civil service of Georgia since its independence. These are divided into five distinct periods. The first one encompasses the time immediately after independence until the adoption of the Law on Civil Service in 1997. The second lasts until the end of the President Shevardnadze’s government as a result of the Rose Revolution in November 2003. The third period – until 2007 – marks important reforms that the new United National Movement (UNM) government was able to implement capitalizing on overwhelming public support it enjoyed. From the second half of 2007 to 2012, the fourth period, the UNM government’s pace of reforms significantly decreased, and even stalled in some areas. The last, fifth period, starts from the end of 2012 and continues to this day. That is when the Georgian Dream Coalition (GD) won the Parliamentary Elections. In the fifth and last period the government showed a clear preference for an independent, Weberian civil service as opposed to New Public Management (NPM) principles preferred by the UNM in their most decisive reforms. Success of reforms irrespective of the type of civil service preferred seems to be highly dependent upon political will and favourable political context.
‘Not safe for discharge’? Words, values, and person-centred care
Authors: Hyslop Brent
Published: December 2019

The phrase ‘not safe for discharge home’ is often heard in relation to an older person in hospital, commonly due to functional limitations or risk of falls. But it remains unclear how such a standard of safety should be set in this context, or who should set it. In addition, labelling someone ‘unsafe’ to return to their own home has significant practical and ethical implications. After briefly exploring these issues, this Commentary suggests that a holistic approach and shared decision-making is required in this setting. Instead of simply declaring someone safe or unsafe for discharge home, specific ‘safety concerns’ (or ‘hazards’) should be identified and addressed as able. Ongoing specific concerns can then be discussed in conjunction with a patient’s values and perceived benefits of returning home, in comparison with potential pros and cons of other discharge options. Overall, this paper suggests that paying attention to our words and values can enhance discharge planning and person-centred care.
Missing data reporting in clinical pharmacy research
Authors: Narayan Sujita W , Yu Ho Kar , Penm Jonathan , Mintzes Barbara , Mirzaei Ardalan , Schneider Carl , Patanwala Asad E
Published: December 2019

Purpose

This study aimed to document the ways by which missing data were handled in clinical pharmacy research to provide an insight into the amount of attention paid to the importance of missing data in this field of research.

Methods

Our cross-sectional descriptive report evaluated 10 journals affiliated with pharmacy organizations in the United States, Canada, the United Kingdom, and Australia. Randomized controlled trials, cohort studies, case-control studies, and cross-sectional studies published in 2018 were included. The primary outcome measure was the proportion of studies that reported the handling of missing data in their methods or results.

Results

A total of 178 studies were included in the analysis. Of these, 19.7% (n = 35) mentioned missing data either in their methods (3.4%, n = 6), results (15.2%, n = 27), or in both sections (1.1%, n = 2). Only 4.5% (n = 8) of the studies mentioned how they handled missing data, the most common method being multiple imputation (n = 3), followed by indicator (n = 2), complete case analysis (n = 2), and simple imputation (n = 1). One study using multiple imputation and both studies using an indicator method also combined other strategies to account for missing data. One study only used complete case analysis for subgroup analysis, and the other study only used this method if a specific baseline variable was missing.

Conclusions

Very few studies in clinical pharmacy literature report any handling of missing data. This has the potential to lead to biased results. We advocate that researchers should report how missing data were handled to increase the transparency of findings and minimize bias.
Quilting Sutures in Rhytidectomy: A Systematic Review of the Literature
Authors: Ballan Anthony , Jabbour Samer , El Rayess Youssef , Jabbour Khalil , El Hachem Lena , Nasr Marwan
Published: December 2019

Background

Different technical variations exist for the utilization of quilting sutures (QS) in rhytidectomy. No systematic review or meta-analysis of the studies describing the use of QS in facelifts has been published to date to the authors’ knowledge.

Objectives

The objective of this study was to summarize all the published data regarding the utilization of QS in rhytidectomy, compare QS techniques, and evaluate their effect on postoperative complications.

Methods

On April 1, 2019, a systematic search of the Medline, Embase, and Cochrane databases was conducted. All the studies describing the usage of QS in facelifts were included in this review. Studies reporting hematoma rate in a QS group and a control group were included in the meta-analysis part of this study.

Results

The initial search of the databases yielded 93 results. Four trials were included in the systematic review and 2 were included in the meta-analysis. The total number of included patients with QS was 527. Two studies employed internal QS and the remaining studies utilized external QS. The meta-analysis found a lower rate of hematoma in the QS group (relative risk, 0.02; 95% confidence interval = 0.00-0.13; P < 0.0001).

Conclusions

QS can be applied either internally or externally and are very effective in reducing hematomas after facelifts. QS could be a great asset in facelifts but should be utilized with caution because additional work is needed to confirm their safety and efficacy.

Level of Evidence: 2
Genetic merit for fertility alters the bovine uterine luminal fluid proteome†
Authors: Gegenfurtner Katrin , Fröhlich Thomas , Flenkenthaler Florian , Kösters Miwako , Fritz Sébastien , Desnoës Olivier , Le Bourhis Daniel , Salvetti Pascal , Sandra Olivier , Charpigny Gilles , Mermillod Pascal , Lonergan Patrick , Wolf Eckhard , Arnold Georg J
Published: December 2019

Over the last decades, fertility of dairy cows has declined due to selection strategies focusing on milk yield. To study the effect of genetic merit for fertility on the proteome of the bovine uterine luminal fluid, Holstein heifers with low- and two groups of heifers with high-fertility index (high-fertility Holstein and Montbéliarde) were investigated. To focus on the maternal effect, heifers from all groups were synchronized and received on Day 7 high-quality embryos. Uterine luminal fluid from Day 19 pregnant heifers was analyzed in a holistic proteomic approach using nano-LC-MS/MS analysis combined with a label-free quantification approach. In total, 1737 proteins were identified, of which 597 differed significantly in abundance between the three groups. The vast majority of proteome differences was found comparing both high-fertility groups to the low-fertility Holstein group, showing that the genetic predisposition for fertility is prevalent regarding the uterine luminal fluid proteome. Evaluation of this dataset using bioinformatic tools revealed an assignment of higher abundant proteins in low-fertility Holstein to several metabolic processes, such as vitamin metabolic process, which comprises folate receptor alpha (FOLR1) and retinol-binding protein, indicating an involvement of disturbed metabolic processes in decreased fertility. Moreover, immune system-related proteins — lactotransferrin and chromogranin A — were enriched in low-fertility cows together with interferon tau 3 h and interferon tau-2. Our results indicate that the genetic merit for fertility leads to substantial quantitative differences at the level of proteins in uterine fluid of pregnant animals, thus altering the microenvironment for the early conceptus.
Developing the Emergency Department Elder Mistreatment Assessment Tool for Social Workers Using a Modified Delphi Technique
Authors: Elman Alyssa , Rosselli Sarah , Burnes David , Clark Sunday , Stern Michael E , LoFaso Veronica M , Mulcare Mary R , Breckman Risa , Rosen Tony
Published: December 2019

Elder mistreatment is common and has serious consequences. The emergency department (ED) may provide a unique opportunity to detect this mistreatment, with social workers often asked to take the lead in assessment and intervention. Despite this, social workers may feel ill-equipped to conduct assessments for potential mistreatment, due in part to a lack of education and training. As a result, the authors created the Emergency Department Elder Mistreatment Assessment Tool for Social Workers (ED-EMATS) using a multiphase, modified Delphi technique with a national group of experts. This tool consists of both an initial and comprehensive component, with 11 and 17 items, respectively. To our knowledge, this represents the first elder abuse assessment tool for social workers designed specifically for use in the ED. The hope is that the ED-EMATS will increase the confidence of ED social workers in assessing for elder mistreatment and help ensure standardization between professionals.
Clinical and Molecular Epidemiology of Invasive Haemophilus influenzae Serotype a Infections in Utah Children
Authors: Crandall Hillary , Christiansen Jennifer , Varghese Alyssa A , Russon Adam , Korgenski E Kent , Bengtson Erika K , Dickey Mandy , Killpack Jarrett , Knackstedt Elizabeth D , Daly Judy A , Ampofo Krow , Pavia Andrew T , Blaschke Anne J
Published: December 2019

Background

Following widespread use of the Haemophilus influenzae serotype b (Hib) vaccine, H. influenzae serotype a (Hia) has emerged as an important pathogen in children in some regions. We describe the clinical features and molecular epidemiology of invasive Hia disease in children in Utah over an 11-year period.

Methods

We identified cases of invasive Hia disease, defined as detection of Hia from a normally sterile site, in children aged <18 years from Utah between 2007 and 2017. Medical records were reviewed to determine demographic characteristics and clinical outcomes. Available Hia isolates were genotyped using multilocus sequence typing, and phylogenetic division was determined using sodC polymerase chain reaction. Presence of the putative virulence-associated IS1016-bexA duplication-deletion was evaluated.

Results

We identified 51 children with invasive Hia. The average annual incidence was 1.7 cases per 100 000 children aged <5 years; 4.8 cases per 100 000 children aged <1 year. The median age was 11.3 months. The most common clinical presentation was meningitis (53%), followed by pneumonia (14%) and septic arthritis (14%). Twenty-two children (43%) required admission to an intensive care unit; 1 died. Sequence type (ST) 62, phylogenetic division II isolates caused 75% (21/28) of disease. No isolates contained the virulence-associated IS1016-bexA duplication-deletion.

Conclusions

Hia is a significant cause of severe invasive bacterial infection in Utah. The majority of infections were caused by ST62 isolates, a phylogenetic division II Hia type that lacks the IS1016-bexA duplication-deletion. Hia ST62 has not been commonly reported elsewhere, suggesting a unique molecular epidemiology in our population.
Moraxella lacunata Subacute Osteomyelitis in a Child
Authors: Mehmeti Fortesa , Cochard Blaise , Cuérel Céline , De Coulon Geraldo , Dayer Romain , Steiger Christina , Ceroni Dimitri
Published: December 2019

Moraxella lacunata is a rare coccobacillus associated with eye and upper respiratory tract infections. It may also have an affinity for bone and joint tissue. We report on 1 case of subacute osteomyelitis of the patella due to M. lacunata that presented as an osteolytic bone lesion in a child.
A novel approach to the computation of one-loop three- and four-point functions. III. The infrared divergent case
Authors: Guillet J Ph. , Pilon E , Shimizu Y , Zidi M S
Published: December 2019

This article is the third and last of a series presenting an alternative method for computing the one-loop scalar integrals. It extends the results of the first two articles to the infrared divergent case. This novel method enjoys a couple of interesting features as compared with the methods found in the literature. It directly proceeds in terms of the quantities driving algebraic reduction methods. It yields a simple decision tree based on the vanishing of internal masses and one-pinched kinematic matrices, which avoids a profusion of cases. Lastly, it extends to kinematics more general than the physical, e.g. collider processes, relevant at one loop. This last feature may be useful when considering the application of this method beyond one loop using generalized one-loop integrals as building blocks.
Plumericin prevents intestinal inflammation and oxidative stress in vitro and in vivo
Authors: Rapa Shara F. , Waltenberger Birgit , Di Paola Rosanna , Adesso Simona , Siracusa Rosalba , Peritore Alessio F. , D'Amico Ramona , Autore Giuseppina , Cuzzocrea Salvatore , Stuppner Hermann , Marzocco Stefania
Published: December 2019

Inflammatory bowel diseases (IBDs) are characterized by an inflammatory and oxidative stress condition in the intestinal tissue. In this study, we evaluated the effect of plumericin, one of the main bioactive components of Himatanthus sucuuba (Woodson) bark, on intestinal inflammation and oxidative stress, both in vitro and in vivo. The effect of plumericin (0.5‐2 µM) in vitro was evaluated in rat intestinal epithelial cells (IEC‐6) treated with lipopolysaccharides from E. coli (10 μg/mL) plus interferon‐γ (10 U/mL). Moreover, a 2,4,6‐dinitrobenzene sulfonic acid (DNBS)‐induced colitis model was used to evaluate the anti‐inflammatory and antioxidant activity of plumericin (3 mg/kg) in vivo. The results showed that plumericin significantly reduces intestinal inflammatory factors such as tumor necrosis factor‐α, cyclooxygenase‐2 and inducible nitric oxide synthase expression, and nitrotyrosine formation. Plumericin also inhibited nuclear factor‐κB translocation, reactive oxygen species (ROS) release, and inflammasome activation. Moreover, plumericin activated the nuclear factor erythroid‐derived 2 pathway in IEC‐6. Using the DNBS‐induced colitis model, a significant reduction in the weight loss and in the development of the macroscopic and histologic signs of colon injury, together with a reduced inflammatory and oxidative stress state, were observed in plumericin‐treated mice. These results indicate that plumericin exerts a strong anti‐inflammatory and antioxidant activity. Thus, it might be a candidate for the development of a new pharmacologic approach for IBDs treatment.
Pulmonary Delivery of Metoprolol Reduces Ventricular Rate During Atrial Fibrillation and Accelerates Conversion to Sinus Rhythm
Authors: Marum Alexandre A. , Araujo Silva Bruna , Bortolotto Alexandre L. , Pedreira Giovanna C. , Tessarolo Silva Fernanda , Medeiros Sofia A. , Nearing Bruce D. , Belardinelli Luiz , Verrier Richard L.
Published: December 2019

Background:

Safe, effective pulmonary delivery of cardioactive agents in humans is under development.

Objectives:

We examined whether intratracheal delivery of metoprolol can reduce ventricular rate during atrial fibrillation (AF) and accelerate conversion to sinus rhythm.

Methods:

In 7 closed-chest, anesthetized Yorkshire pigs, AF was induced by intrapericardial infusion of acetylcholine (1 mL of 102.5-mM solution) followed by atrial burst pacing and was allowed to continue for 2 minutes before intratracheal instillation of sterile water or metoprolol (0.2-mg/kg bolus) using a catheter positioned at the bifurcation of the main bronchi. High-resolution electrograms were obtained from catheters fluoroscopically positioned in the right atrium and left ventricle.

Results:

Rapid intratracheal instillation of metoprolol caused a 32-beat/min reduction in ventricular rate during AF (from 272 ± 13.7 to 240 ± 12.6 beats/min, P = 0.008) and a 2.3-minute reduction in AF duration (from 10.3 ± 2.0 to 8.0 ± 1.4 minutes, P = 0.018) compared with sterile water control. Conversion of AF to sinus rhythm was associated with rapid restoration (5–6 minutes) of heart rate and arterial blood pressure toward control values. Intratracheal metoprolol reduced AF dominant frequency by 31% (from 8.7 ± 0.9 to 6.0 ± 1.1 Hz, P = 0.04) compared with control and resulted in a trend toward a 5% increase in PR interval (from 174 ± 11.2 to 182 ± 11.4 ms, P = 0.07).

Conclusions:

Intratracheal delivery of metoprolol effectively reduces ventricular rate during AF and accelerates conversion to normal sinus rhythm in a pig model of acetylcholine-induced AF.
Cefazolin Monotherapy Versus Cefazolin Plus Aminoglycosides for Antimicrobial Prophylaxis of Type III Open Fractures
Authors: Patanwala Asad E. , Radosevich John J. , Meshay Ian , Naderi Mandana , Culver Mark A. , Lee Yong G. , Weinberg Jordan A. , Khobrani Moteb , Nix David E.
Published: December 2019

Background:

There are conflicting recommendations between organizations regarding aminoglycoside use for the prophylaxis of type III open fractures.

Study Question:

To compare cefazolin monotherapy versus cefazolin plus aminoglycoside therapy for prophylaxis of type III open fractures in trauma patients.

Study Design:

This was a multicenter, retrospective, cohort study conducted in 3 academic medical centers in the United States. Consecutive adult trauma patients with type III open fractures between January 2014 and September 2016 were included. Patients were divided into 2 groups: (1) cefazolin monotherapy versus (2) cefazolin plus aminoglycoside.

Measures and Outcomes:

The primary outcome measure was the occurrence of infection at the open fracture site. The secondary outcome measure was the occurrence of acute kidney injury.

Results:

There were 134 patients included in the study cohort. Of these, 39 received cefazolin monotherapy and 95 received cefazolin plus aminoglycoside. Overall, the mean age was 39 ± 15 years, 105 (78%) were male, and the most common fracture location was tibia/fibula (n = 74, 56%). Infection at the open fracture site occurred in 6 of 39 patients (15%) in the cefazolin monotherapy group and 15 of 95 patients (16%) in the cefazolin plus aminoglycoside group (P = 1.000). Acute kidney injury occurred in 0 of 39 (0%) in the cefazolin monotherapy group and 1 of 95 (1%) in the cefazolin plus aminoglycoside group (P = 1.000).

Conclusions:

Cefazolin monotherapy may be appropriate for antimicrobial prophylaxis of type III open fractures in trauma patients.
The Impact of Celiac Disease on Caregivers’ Well‐being
Authors: Satherley Rose‐Marie , Coburn Shayna S. , Germone Monique
Published: December 2019

Objectives:

A diagnosis of celiac disease (CD) requires individuals to adopt a strict gluten‐free diet. As children with CD must rely on their caregivers for guidance and support with managing the gluten‐free diet, CD may challenge the caregiver's emotional and social well‐being. The primary objective of this mixed‐methods systematic review was to synthesize research investigating the impact of CD on caregiver's well‐being.

Methods:

Five databases were systematically searched from 1990 to 2018 to identify all empirical studies that assessed well‐being in caregivers of children (0–18 years) with CD. Qualitative and quantitative data were extracted separately before being integrated to explore key themes across the studies.

Results:

Twelve studies were identified that explored the well‐being of caregivers of children with CD (3 qualitative, 9 quantitative), reporting on 665 caregivers. The quality of evidence was limited across studies. Synthesis of results revealed 3 themes (Caregiver Responsibility, Caregiver Well‐Being and Concern for Child's Health, Implications for the Family) describing the impact of a child with CD on caregiver well‐being.

Conclusions:

Caregivers of children with CD may experience difficulties that impact their well‐being; specific difficulties identified included the impact of caregivers’ social activities, finances, and anxiety. The findings detailed in this review point toward factors that may guide health care personnel to provide support for the caregivers of children with CD.
Drugs in Focus
Authors: Pienar Corina , Benninga Marc A. , Broekaert Ilse J. , Dolinsek Jernej , Mas Emmanuel , Miele Erasmo , Orel Rok , Ribes‐Koninckx Carmen , Thomassen Rut‐Anne , Thomson Mike , Tzivinikos Christos , Thapar Nikhil
Published: December 2019

Acute diarrhoea is a leading cause of morbidity and mortality in the paediatric population. Racecadotril is an antisecretory drug recommended as an adjuvant antidiarrhoeal treatment.

In the small bowel, the enzyme neutral endopeptidase (NEP) inhibits the action of enkephalins, which prevent water and electrolyte hypersecretion. By inhibiting NEP, racecadotril allows enkephalins to exhibit their antisecretory effects. Consequently, racecadotril reduces the secretion of water and electrolytes in the small intestine, without having an effect on intestinal motility. No serious adverse events related to racecadotril have been reported.

Racecadotril has proven its efficacy as an adjuvant antidiarrhoeal drug with a good safety profile. Its addition to oral rehydration solution (ORS) appears clinically beneficial and potentially leads to health care savings.
Online couple interventions in cancer
Authors: Vanstone Ruth , Fergus Karen D.
Published: December 2019

Purpose of review

Cancer diagnosis and treatment can have long-lasting psychological and physical consequences that affect both patients and their intimate partners. Improved understanding of extant dyadic interventions in the context of cancer, and how access to these may be enhanced through web-based technologies, introduce new directions for how cancer-related psychological distress for couples may be ameliorated.

Recent findings

Couples are negatively impacted by cancer, both individually, and as a dyad. Bolstering techniques to support effective communication about common cancer-related concerns and support for adjusting to new roles and responsibilities may help to strengthen the couple's relationship so partners are better able to cope with cancer. Although there are various intervention options available for couples dealing with cancer, many pose barriers to participation because of constraints on time and/or distance. However, online interventions have been shown to be effective, both in easing psychological distress and reducing participant burden.

Couples dealing with cancer experience psychological distress and must learn to navigate changing roles and responsibilities in the face of the disease. Online interventions offer flexible and innovative platforms and programs that help to address couples’ educational needs while strengthening dyadic coping.
The AP-2 endocytic adaptor complex in Candida albicans morphology and virulence
Authors: Knafler Harriet , Rooij Iwona Smaczynska-de , Ayscough Kathryn
Published: December 2019

Fungi such as Candida species are a major cause of hospital-acquired infections especially in immuno-compromised patients, and invasive candidiasis is associated with a high mortality rate. Central to Candida albicans virulence is its ability to switch between budding (yeast) and filamentous (hyphal) growth, and to colonise different body niches. In each distinct environment it must remodel its cell surface to ensure appropriate levels of transporters and cell wall biosynthesis enzymes. Endocytosis is known to be a critical pathway in surface remodelling, allowing cells to internalise proteins that are no longer needed at the plasma membrane. Endocytosis is also crucial to highly polarised hyphal growth, where endocytic recycling of key membrane proteins is essential to maintain their polarised location at the hyphal tip. The aim of this study is to investigate the role of the AP-2 endocytic adaptor complex in endocytosis within C. albicans. Homozygous deletions were generated in an essential subunit of the AP-2 complex. The deletion did not affect rates of cell growth or fluid phase endocytosis, but using fluorescence and electron microscopy, defects were observed in hyphal polarisation and in cell wall organisation. We have shown that the AP-2 complex is required for the recycling of the key cell wall biosynthesis enzyme Chs3, via its Yxxφ internalisation motif(s). We demonstrate this interaction is critical for correct cell wall deposition and polarised growth. Thus, AP-2 mediated endocytic recycling is a key step in the regulation of the fungal cell wall.
Using Saccharomyces cerevisiae as a tool for mapping genetic variants underlying differences in response to anti-cancer agents
Authors: Almayouf Salwa , Barton David B.H. , Hu Yue , Foster Steven , Louis Ed J.
Published: December 2019

Background

There are variations in response to anti-cancer agents amongst individuals because they are complex traits controlled by multiple genes and environmental factors. Here, we aimed to decipher the genes controlling the variations in response to aspirin, metformin, disulfiram and 5-fluorouracil using quantitative trait loci (QTL) mapping in yeast before translating findings to humans. This approach was feasible due to the conservation of genes between these two organisms.

Method

A panel of 111 F12 meiotic segregants generated from a four-parent S. cerevisiae cross were genotyped by whole genome sequencing. Segregants growth in different treatments were phenotyped using PHENOS. Subsequently, linkage-based fine QTL mapping was performed to locate regions of the genome and the identification of causative genes.

Results

Linkage analysis has mapped hundreds of genetic loci in the yeast genome responsible for the variations in response to the agents tested. Conserved homologs to human genes were identified. Some hits have been previously supported in the literature such as the effect of aspirin and metformin on MTOR thus validating this screening approach. Novel genes identified and pathway enrichment revealed mechanisms by which these agents may exhibit their anti-cancer properties.

Conclusion

Detection of genetic variants influencing the differences in drug response could help identify individuals at risk or benefit of using anti-cancer agents. Current work includes validating the alleles of causative genes (Rad57 and VMA22) in yeast by reciprocal hemizygosity and allele swapping. This study could aid the development of biomarkers for drug response and validate the repurposing of drugs for cancer prevention.