Assessment of mothers’ satisfaction with health care during childbirth in a tertiary-level maternity ward

Aim To evaluate satisfaction of mothers who gave birth at term with received hospital care and to find areas for improvement at a tertiary hospital. Methods A cross-sectional study at the Department of Obstetrics & Gynaecology at the University Clinical Hospital Mostar was conducted by an anonymous survey using a questionnaire designed exclusively for this study. A total of 100 mothers were included in the study. Results Satisfaction with midwives’ communication and their approach to the women during their stay in the delivery room was rated significantly higher (4.7±0.6) when compared to obstetricians-gynaecologists (4.5±0.8) (p=0.02). Midwives were rated better in providing breastfeeding information (4.5±0.8) than for the speed of arrival after a call bell (average grade 4.2±1.0). Respondents were least satisfied with the hygiene (toilet, shower and rooms) and the quality of food (average grades 3.8±1.1 and 3.9±1.0, respectively). Mothers with previous experience in childbirth at the same hospital rated current stay with a similar level of satisfaction. Conclusion Good communication skills of medical and non-medical staff are a recommended step to maintain mothers′ childbirth satisfaction, while improvement in quality of nutrition and hygiene should be mandatory

Activation of Innate and Adaptive Immunity by a Recombinant Human Cytomegalovirus Strain Expressing an NKG2D Ligand.

Development of an effective vaccine against human cytomegalovirus (HCMV) is a need of utmost medical importance. Generally, it is believed that a live attenuated vaccine would best provide protective immunity against this tenacious pathogen. Here, we propose a strategy for an HCMV vaccine that aims at the simultaneous activation of innate and adaptive immune responses. An HCMV strain expressing the host ligand ULBP2 for the NKG2D receptor was found to be susceptible to control by natural killer (NK) cells, and preserved the ability to stimulate HCMV-specific T cells. Infection with the ULBP2-expressing HCMV strain caused diminished cell surface levels of MHC class I molecules. While expression of the NKG2D ligand increased the cytolytic activity of NK cells, NKG2D engagement in CD8+ T cells provided co-stimulation and compensated for lower MHC class I expression. Altogether, our data indicate that triggering of both arms of the immune system is a promising approach applicable to the generation of a live attenuated HCMV vaccine

Inflammatory monocytes and NK cells play a crucial role in DNAM-1-dependent control of cytomegalovirus infection

The poliovirus receptor (PVR) is a ubiquitously expressed glycoprotein involved in cellular adhesion and immune response. It engages the activating receptor DNAX accessory molecule (DNAM)-1, the inhibitory receptor TIG IT, and the CD96 receptor with both activating and inhibitory functions. Human cytomegalovirus (HCMV) down-regulates PVR expression, but the significance of this viral function in vivo remains unknown. Here, we demonstrate that mouse CMV (MCMV) also down-regulates the surface PVR. The m20.1 protein of MCMV retains PVR in the endoplasmic reticulum and promotes its degradation. A MCMV mutant lacking the PVR inhibitor was attenuated in normal mice but not in mice lacking DNAM-1. This attenuation was partially reversed by NK cell depletion, whereas the simultaneous depletion of mononuclear phagocytes abolished the virus control. This effect was associated with the increased expression of DNAM-1, whereas TIG IT and CD96 were absent on these cells. An increased level of proinflammatory cytokines in sera of mice infected with the virus lacking the m20.1 and an increased production of iNOS by inflammatory monocytes was observed. Blocking of CCL2 or the inhibition of iNOS significantly increased titer of the virus lacking m20.1. In this study, we have demonstrated that inflammatory monocytes, together with NK cells, are essential in the early control of CMV through the DNAM-1–PVR pathwa

Superior induction and maintenance of protective CD8 T cells in mice infected with mouse cytomegalovirus vector expressing RAE-1

Due to a unique pattern of CD8 T-cell response induced by cytomegaloviruses (CMVs), live attenuated CMVs are attractive candidates for vaccine vectors for a number of clinically relevant infections and tumors. NKG2D is one of the most important activating NK cell receptors that plays a role in costimulation of CD8 T cells. Here we demonstrate that the expression of CD8 T-cell epitope of Listeria monocytogenes by a recombinant mouse CMV (MCMV) expressing the NKG2D ligand retinoic acid early-inducible protein 1-gamma (RAE-1γ) dramatically enhanced the effectiveness and longevity of epitope-specific CD8 T-cell response and conferred protection against a subsequent challenge infection with Listeria monocytogenes. Unexpectedly, theattenuatedgrowth in vivo of the CMV vector expressing RAE-1γ and its capacity to enhance specific CD8 T-cell response were preserved even in mice lacking NKG2D, implying additional immune function for RAE-1γ beyond engagement of NKG2D. Thus, vectors expressing RAE-1γ represent a promising approach in the development of CD8 T-cell– based vaccine

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Bis(phenylethynyl)arene Linkers in Tetracationic Bis‐triarylborane Chromophores Control Fluorimetric and Raman Sensing of Various DNAs and RNAs

We report four new luminescent tetracationic bis-triarylborane DNA and RNA sensors that show high binding affinities, in several cases even in the nM range. Three of the compounds contain substituted, highly emissive, and structurally flexible bis(2, 6-dimethylphenyl-4- ethynyl)arene linkers (3: arene = 5, 5’-2, 2’-bithiophene ; 4: arene = 1, 4-benzene ; 5: arene = 9, 10-anthracene) between the two boryl moieties and serve as efficient dual Raman and fluorescence chromophores. The shorter analogue 6 employs bis(2, 6- dimethylphenyl)-9, 10-anthracene as the linker and demonstrates the importance of an adequate linker length with a certain level of flexibility by exhibiting generally lower binding affinities than 3-5. Pronounced aggregation - deaggregation processes are observed in fluorimetric titration experiments with DNA for compounds 3 and 5. Molecular modelling of complexes of 5 with AT-DNA, suggest the minor 2 groove as the dominant binding site for monomeric 5, but demonstrate that dimers of 5 can also be accommodated. Strong SERS responses for 3-5 (particularly compound 5) vs. a very weak response for 6 demonstrate the importance of triple bonds for strong Raman activity in molecules of this compound class. The energy of the characteristic stretching vibration of the C≡C bonds is significantly dependent on the aromatic moiety between the triple bonds. The insertion of aromatic moieties between two C≡C bonds thus offers an alternative design for dual Raman and fluorescence chromophores, applicable in multiplex biological Raman imaging

Bis(phenylethynyl)arene Linkers in Tetracationic Bis-triarylborane Chromophores Control Fluorimetric and Raman Sensing of Various DNAs and RNAs

We report four new luminescent tetracationic bis-triarylborane DNA and RNA sensors that show high binding affinities, in several cases even in the nanomolar range. Three of the compounds contain substituted, highly emissive and structurally flexible bis(2,6-dimethylphenyl-4-ethynyl)arene linkers (3: arene=5,5′-2,2′-bithiophene; 4: arene=1,4-benzene; 5: arene=9,10-anthracene) between the two boryl moieties and serve as efficient dual Raman and fluorescence chromophores. The shorter analogue 6 employs 9,10-anthracene as the linker and demonstrates the importance of an adequate linker length with a certain level of flexibility by exhibiting generally lower binding affinities than 3–5. Pronounced aggregation–deaggregation processes are observed in fluorimetric titration experiments with DNA for compounds 3 and 5. Molecular modelling of complexes of 5 with AT-DNA, suggest the minor groove as the dominant binding site for monomeric 5, but demonstrate that dimers of 5 can also be accommodated. Strong SERS responses for 3–5 versus a very weak response for 6, particularly the strong signals from anthracene itself observed for 5 but not for 6, demonstrate the importance of triple bonds for strong Raman activity in molecules of this compound class. The energy of the characteristic stretching vibration of the C≡C bonds is significantly dependent on the aromatic moiety between the triple bonds. The insertion of aromatic moieties between two C≡C bonds thus offers an alternative design for dual Raman and fluorescence chromophores, applicable in multiplex biological Raman imaging

T cell activation with ULBP2-TB40 infected DC is not impaired in the presence of NK cells.

<p>Intracellular cytokine staining (IFNγ/TNFα or IFNγ) and degranulation (CD107a staining) of (A) NK cells (CD3-CD56+) and (B) CD8+ T cells (CD3+CD8+) upon co-cultivation of PBMC from one donor with autologous monocyte-derived DC treated or infected as indicated. Graphs on the right are compiled data of experiments performed with cells of 4 HCMV-seropositive donors. Data generated with cells of individual donors are connected by lines and are representative of one of three independent experiment performed. Statistical analysis was done with one-way ANOVA Friedman test followed by Dunn’s Multiple Comparison test. *, P < 0.05.</p

Spread of the ULBP2-expressing HCMV strain is strongly controlled by NK cells.

<p>(A) The ULBP2-TB40 strain was generated by insertion of a ULBP2 expression cassette (driven by the mouse CMV major immediate-early promoter (mMIEP)), replacing ORF UL16 in the BAC-cloned TB40 genome that lacks also the US1 to US6 region. (B) HFF were infected at an MOI of 1 with TB40 or ULBP2-TB40 or left uninfected. The infection rate at day 1 p.i. was determined by flow cytometry following intranuclear staining with an IE1/2-specific antibody. Lower panels, ULBP2 surface expression on uninfected, TB40 (black lines) or ULBP2-TB40 infected cells (red line). Grey fill, isotype control. (C) Cytotoxicity assays were set up at the indicated effector/target (E/T) ratios with primary NK cells and uninfected (white circles), TB40 (black circles) and ULBP2-TB40 (red circles) infected HFF (MOI 1, 1 d.p.i.). Results are means ± SEM of 3 independent experiments performed with NK cells of one donor. (D) Compiled data of cytotoxicity assays performed with NK cells from 3 HCMV-negative donors as in (C), and in addition with ULBP2-TB40 infected target cells in the presence of an NKG2D blocking antibody (aNKG2D) or isotype antibody (iso). Results are medians (with range) of specific lysis at E/T ratio 8:1 (n = 3 donors). Data were compiled from 3 independent experiments. (E) Focus expansion assays using TB40 or ULBP2-TB40 infected HFF and primary NK cells. At day 3 of co-culturing infected cells were detected by fluorescence microscopy following staining with an IE1/2 antibody. Graph displays cumulative data as medians (with range) of experiments done with NK cells of 6 donors. A focus of infection is defined as a cluster of 3 to 60 infected cells. Four wells were analyzed per donor and all infectious foci were counted with 2 to 8 foci being present per well. Images are from one representative experiment. (F) Focus expansion assays were carried out as described in (E) and additionally in the presence of NKG2D blocking (aNKG2D) or isotype antibodies (iso) using NK cells from 4 donors. Statistical analysis for (C) (NK cell-mediated cytotoxicity assay at E/T ratio 16:1) was done with two-way ANOVA with Bonferroni correction and for (E, F) with Mann-Whitney t-test. *, P < 0.05, **, P < 0.01, ***, P < 0.001.</p

<i>Ex vivo</i> expansion with ULBP2-TB40 infected DC results in highly functional HCMV-specific CD8+ T cells.

<p>(A) Dot plots of an experiment performed with cells from one donor indicating percentage of pp65-specific CD8+ T cells before expansion and after 14 days of co-culture with TB40 or ULBP2-TB40 infected DC. Graph displays compiled data for four donors as percentages of pp65-specific CD8+ T cells expanded with uninfected (white circles), TB40 (black circles) and ULBP2-TB40 (red circles) infected DC. Data obtained with cells from individual donors are connected by lines. (B) Cytokine production and degranulation of pp65-specific CD8+ T cells (obtained as described in (A) or upon co-culture with pp65 peptide-loaded DC (pp65-mDC)) in response to pp65 peptide-loaded DC as target cells. Representative staining for IFNγ, TNFα and CD107a of pp65-specific CD8+ T cells obtained from one donor. The graphs at right are compiled data for 4 donors. (C) Multifunctionality of the expanded pp65-specific CD8+ T cells. Multifunctional subsets were determined using Boolean analysis based on the simultaneous measurement of IFNγ, TNFα and CD107a positive cells. Pie charts denote the proportion of pp65-specific CD8+ T cells having 1, 2 or 3 functions for 4 different donors. Data are representative of one out of two independent experiment performed. Statistical analysis was done with one-way ANOVA Friedman test followed by Dunn’s Multiple Comparison test. *, P < 0.05.</p