The blot was blocked with 10% skim milk solution for 2 hours. After washing with phosphate-buffered saline (PBS) solution, the blot was probed overnight using a polyclonal flagellar antibody raised in a rabbit against isolated flagellar filaments [41]. Protein A-alkaline phosphatase (Sigma-Aldrich) was used as the secondary antibody. The blot was washed with PBS and was developed using NBT/BCIP (Sigma). Preparation of samples for tandem mass spectrometry analysis (MS/MS) The flagellar protein samples were run on a polyacrylamide gel as described above. Staining and destaining of the protein gel were performed following standard protocols

[42]. The gel was soaked overnight in a staining solution containing 0.1% Coomassie Brilliant Blue (R-250; Sigma), 40% methanol, and 10% acetic acid. Destaining was done using a solution containing 40% GW572016 methanol and 10% acetic

acid. The bands (between approximately 25-37kDa) HKI-272 datasheet were excised and submitted to the Southern Alberta Mass Spectrometry (SAMS) Centre at the University of Calgary check details for LC-MS/MS analysis. Two bands within the size range were observed in the gel. The two bands were analyzed separately for 3841 and in combination for VF39SM. The gel slices were rinsed once with HPLC-grade water and then twice with 25 mM ammonium bicarbonate in 50% (v/v) acetonitrile. The gel slices were dehydrated with acetonitrile prior to lyophilization. The dehydrated gel was resuspended in 25 mM ammonium bicarbonate (pH8.0) and samples were digested with trypsin. The peptides were extracted from the gel using 1% formic acid in 50% acetonitrile. The extracts were reduced to dryness and then reconstituted in mobile phase of the buffer (3% acetonitrile with 0.2% formic acid) for liquid chromatography. Tandem mass spectrometry analysis (MS/MS) The digests were analyzed using

an integrated Agilent 1100 LC-Ion-Trap-XCT-Ultra system (Agilent Technologies, Santa Clara, CA), which has an integrated Montelukast Sodium fluidic cartridge for peptide capture, separation, and nano-spraying (HPLC Chip). The injected samples were trapped and desalted for 5 minutes using a pre-column channel (40-nl volume; Zorbax 300 SB-C18) with an auxiliary pump that delivers 3% acetonitrile and 0.2% formic acid at a flowrate of 4 μl/minute. The peptides were reverse-eluted from the trapping column and separated on a 150 mm-long analytical column (Zorbax 300SB-C18) at a flowrate of 0.3 μl/minute. The peptides were eluted using a 5-70% (v/v) acetonitrile gradient in 0.2% (v/v) formic acid over a period of 10 minutes. The MS/MS spectra were collected by data-dependent acquisition, with parent ion scans of 8100 Th/s over m/z 400-2,000. MS/MS scans at the same rate over m/z 100-2200. Mass Spectrometry Data Analysis DataAnalysis software for the 6300 series ion trap, v3.4 (build 175) was used to extract the peak-list data. The MS/MS data were analyzed using Mascot v2.

Also we have gained additional experience with the use of HBO therapy for severe life-threatening infections such as clostridial myonecrosis and other aerobic and anaerobic NSTI. Regardless of the type of surgical strategy applied, the HBO therapy should never delay the emergency of the surgical intervention, including the treatment of Clostridium perfrigens causing gas gangrene [36, 54, 57]. Reconstructive surgery The reconstruction of skin defects either on the A-1155463 in vitro extremities and torso, or on the abdominal or chest wall, should be performed using several different techniques and surgical materials on each patient.

As is often seen, a complete loss of skin or dermal structures needs a complex, multilayer reconstruction especially in functional areas of the body and on the extremities. Novel concepts of layer-specific reconstruction include biologic meshes, which are an alternative

to flap and skin graft surgery, especially in abdominal and chest wall reconstructions [58–61]. After the wound stabilizes and fresh granulation tissue without any signs of acute infection we perform staging reconstructions using simple to complex reconstructive methods. Epigenetics inhibitor The main contributing factor for reconstructive method-selection was the extent and the localization of the defect and the patient’s condition [51–53]. Topical negative pressure therapy has been reported to remove exudates, cover wounds securely, stimulate angiogenesis [6, 49] and reduce bacterial contamination [50]. It also reduces the surface area of the wound, improves the rate of granulation tissue formation, reduces the number of surgical

excision procedures needed, as well as enables better healing performance of skin grafts and biologic meshes. The cost benefit of that novel therapy is evident, but the complications of TNP still exist and include damage to surrounding tissue due to pressure effects, pain during dressing changes and discomfort because of very Sirolimus bulky dressing [52]. Newer data recommend the use of TNP in the acute traumatic military settings [58]. Leininger at all used TNP in the deployed military settings (at R3 stage of-NATO medical care) where they treated all Iraqi selleck casualties with TNP dressing after their first debridement (77 cases) [59]. They reported that infection rates dropped from 81% to 0% after using the TNP management strategy. Our experience has shown the use of this wound management technique to remove exudates, improve the patient comfort, reduce the wound size and the time for wound stabilization, to allow the formation of fresh granulation tissue, and better healing of skin grafts and flaps [36].

All authors VX-765 manufacturer have given

final approval of the version to be published.”
“Background Physical activity leads to increased metabolic rate and heat production [1], resulting in loss of water and electrolytes and glycogen depletion in the liver and muscles [1, 2]. The loss of these elements may lead to dehydration, affecting physical performance and impairing health [3]. Fluid replacement using isotonic solution may attenuate or prevent many metabolic, cardiovascular, thermoregulatory and performance perturbations [4, 5]. Moreover, according to Brouns et al., [6] and Coyle [7], sports drinks without caffeine can help to maintain physiological homeostasis. Another aspect of risk related to AZD6244 supplier exercise is failure

of cardiovascular function, especially for practitioners who exercise infrequently [8]. It is known CB-839 that reduced cardiac parasympathetic regulation associated with increased sympathetic activation may trigger malignant ventricular arrhythmias, and that systemic metabolic disorders (electrolyte imbalance, hypoxia), as well as hemodynamic or neurophysiological (fluctuations in the activity of the autonomic nervous system) disorders appear to play an important role in lethal arrhythmias [9]. In addition, the physiological overload imposed on the body is enhanced when exercise is associated with dehydration. According to Carter et al., [5], “the combination of these two factors suggests changes in the global cardiac autonomic stability”. In combination with dehydration, exercise has been shown to cause post-exercise alterations in the baroreflex control of blood pressure [10]. Charkoudian et al., [10] demonstrated that even modest hypohydration (1.6% of body weight) can blunt baroreceptor control of blood pressure and that physiological responses were not

observed following an intravenous infusion of saline to restore the plasma volume after exercise in the heat. Although it is known that changes in the cardiovascular system are caused by hydration during and after exercise, Cyclin-dependent kinase 3 few studies have evaluated the influence of hydration on the autonomic nervous system (ANS) and none have evaluated this influence when isotonic drink is also administered during and after prolonged exercise. Our purpose, therefore, was to evaluate the effects of hydration protocols on autonomic modulation of the heart in young people during and post-exercise. We hypothesized that hydration during exercise and recovery may attenuate autonomic changes induced by exercise and accelerate recovery.

1998). Fewer studies use the effort–reward imbalance (ERI) model (Siegrist et al. 2004) or the organisational injustice model (Elovainio et al. 2006) or other instruments. There are different ways to derive PAFs for a population (e.g., country or region), either directly from a population-based study or indirectly. With the indirect

approach, risk estimates from one or more analytical studies are retrieved and combined with information on the fraction of exposed persons in the general population from other sources (mainly surveys). Risk estimates may be derived from studies selected based on specific quality criteria (e.g., a certain design and/or statistical model including the relevant confounders) or from meta-analyses, see more respectively. When using this method, survey questions to estimate the prevalence of exposure need to be comparable to the instruments

used for the exposure in the observational studies, which are the basis for the calculation of risk estimates. Validity of the PAF depends heavily on the estimation of the prevalence as well as risk estimates, given that they are correctly estimated (Olsen 1995). Niedhammer et al. (2013) used proxies for the job Lonafarnib cell line strain and effort–reward imbalance from the fourth European Working Condition Survey (EWCS) and combined the prevalences with risk estimates from published meta-analyses. With this indirect method, the authors describe PAFs between 2.51 and 5.77 % for Inositol monophosphatase 1 job strain and 9.78–27.89 % for

the effort–reward ratio >1 in the European countries. Reviewing the literature on fractions of CVD attributable to psychosocial work factors, we also saw that the estimated Fludarabine mouse PAFs differ severely between countries (Backé et al. 2013; Backé and Latza 2013). With the indirect approach, PAFs for cardiovascular outcomes attributed to occupational stress have been derived for the United States (Steenland et al. 2003), Finland (Nurminen and Karjalainen 2001), Korea (Ha et al. 2011), and France (Sultan-Taïeb et al. 2011). For Sweden, PAFs in relation to several diseases were calculated by Järvholm et al. (2013). Here, with respect to job strain and myocardial infarction, calculations with the direct approach were based on a population-based case reference study (Peter et al. 2002). Illustrated for those European countries, where information about PAFs (besides the calculations based on EWCS) are available, PAF estimates differ depending on different prevalence of the exposure but also on different choices in the selection of studies indicating the risk estimates (Table 1). Besides, also discussed by Niedhammer et al. (2013), some authors choose age- and gender-adjusted risk estimates, and some multiple-adjusted risk estimates, respectively. The latter may result in an underestimation of the relative risk when mediators such as high blood pressure or high cholesterol are included. In a recent meta-analysis (Kivimäki et al.

3%) respectively six occasions (15.0%). Accordingly, L. gasseri/iners as members of the normal VMF (n = 83) continued to be present in a following trimester at a rate of 84.3%. Hence, compared to L. crispatus, L. gasseri

and/or iners were found to be significantly less stable BTSA1 order microflora components (McNemar odds ratio 23.33, 95% CI 7.10 – 92.69, p < 0.001). Association between the presence of distinct Lactobacillus species at baseline and vaginal microflora status on follow-up We explored whether the observations on the stability of the grade I VMF as determined by Gram stain correlated with the observations on the stability of the distinct Lactobacillus species observed with grade I VMF as determined through tRFLP and culture. Normal microflora comprising L. crispatus as a member (n = 83) rarely shifted away from grade I VMF microflora (2.4%). Such a shift was observed on merely two occasions click here (shift to grade I-like and grade II VMF respectively) and was not associated with the disappearance of L. crispatus (Table 5). Normal microflora

comprising L. jensenii as a member (n = 42) shifted VX-680 cost away from grade I VMF microflora on three occasions (on each occasion involving a grade Ib VMF to grade II VMF transition) (7.2%), and on two occasions this was associated with the loss of L. jensenii (Table 5). Finally, normal VMF comprising L. gasseri/iners as a member (n = 83) converted to abnormal VMF on twelve occasions (involving conversion from grade I VMF to grade I-like five times, to grade II six times, and to grade III once) (14.5%) (Table 5), which was associated DCLK1 with the disappearance of L. gasseri/iners in merely two out of the twelve normal to abnormal VMF transitions. It may be added that in the aforementioned

instances, including the two L. crispatus comprising VMF and the three L. jensenii comprising VMF which converted to abnormal VMF, L. gasseri/iners was actually present alongside L. crispatus respectively L. jensenii. So, in summary conversion from normal VMF to abnormal VMF was associated twice with grade I L. crispatus + L. gasseri/iners microflora, three times with grade I L. jensenii + L. gasseri/iners microbiota, and seven times with grade I microbiota only containing L. gasseri/iners. In one additional case, conversion from normal VMF to abnormal VMF occurred with a woman with grade Ib VMF from which no lactobacilli could be identified through tRFLP and culture. Table 5 Association between Lactobacillus type as part of grade I microflora (on culture and tRFLP) and microflora status (on Gram stain) on follow-up when accounting for the first-to-second and second-to-third trimester transitions Lactobacillus species (culture and tRFLP) at baseline Gram stain category on follow-up all samples with an L. crispatus TRF (n = 83)      ▪ sustained grade I microflora 97.6% (81)    ▪ shift to an abnormal microflora   – grade I-like 1.2% (1) – grade II 1.2% (1) – grade III – - grade IV – all samples with an L.

These subjects spanned a wide range of ages (20 to 78 years) and anthropometrics (BMI range 17 to 39). For this study, DXA screening was not performed prior to enrollment; therefore, no BMD inclusion/exclusion criteria was used. Copanlisib in vitro For both cohorts, history of or

evidence for metabolic bone Vistusertib concentration disease other than postmenopausal bone loss was an exclusion criterion, as was treatment within the previous year with any compound known to influence bone turnover. Both study protocols were approved by the UCSF Committee of Human Research, and all subjects gave written informed consent prior to participation. HR-pQCT All subjects described below were imaged in a clinical HR-pQCT system (XtremeCT, Scanco Medical AG, Brüttisellen, Switzerland)

using the manufacturer’s standard this website in vivo protocol described in previous patient studies [11, 12, 14]. This system consists of a microfocus X-ray source with a 70-µm focal spot size. The tube voltage was fixed at 60kVp while the current was 900 μA. Filters of 0.3 mm Cu and 1 mm Al are positioned at the aperture to filter soft X-rays in order to reduce patient dose and limit beam-hardening effects. The cone beam X-ray field is incident upon a structured CsI (40 mg/cm2) scintillator coupled by a fiber optic taper to a 2D 3,072 × 256 element CCD detector with a 41-µm pitch. The subject’s forearm was immobilized in a carbon fiber cast that was fixed within the gantry of the scanner. A single dorsal–palmar projection image of the distal radius was acquired to define the tomographic scan region.

This region spans 9.02 mm in length (110 slices) and was fixed starting at 9.5 mm proximal from the mid-jointline and extending proximally (Fig. 1a). For tomography, 750 projections were acquired over 180° with a 100-ms integration time at each angular position. The 12.6-cm field of view was reconstructed across a 1,536 × 1,536 matrix using a modified Feldkamp algorithm, yielding 82 µm voxels [21]. Total scan time was 2.8 min with an equivalent Etomidate dose of approximately 4.2 µSv. Fig. 1 Images indicating the standard ultra-distal ROI for each device; HR-pQCT scout scan (a), Hologic DXA (b), Lunar DXA (c) The reconstructed linear attenuation values were converted to hydroxyapatite (HA) mineral densities using a beam-hardening correction and phantom calibration procedure previously described for an ex vivo microtomography system [22]. The calibration phantom (Scanco Medical AG, Brüttisellen, Switzerland) was composed of five cylinders of HA–resin mixtures with a range of mineral concentrations (0, 100, 200, 400, and 800 mg HA/cm3) where 0 mg HA/cm3 represents a soft tissue equivalent background devoid of mineral. The reconstructed images were segmented using semi-automatically drawn contours at the periosteal surface of the radius. The total vBMD of the radius was calculated as the mean calibrated mineral density within this volume of interest (VOI).

Maartenskliniek Nijmegen; Leiden University Medical Center; University Medical Center Utrecht and Wilhelmina Hospital Assen.” Grants for this study were received from NutsOhra Fonds and Mobiliteitsfonds HBO. The authors are grateful to the subjects

who actively participated in the study and to the students that led the FCE tests. Also they thank Anita Mooij and Annet ter Avest, research nurses of the Medisch Spectrum Twente and the Ziekenhuisgroep Twente, Wim Hilberdink, physical therapist in Groningen, and Janet Wesseling, CHECK-coordinator, for their contributions to the study. Conflict of interest statement The authors declare that they have CFTRinh-172 no conflict of interest. Open Access This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. References Altman R, Asch E, Bloch D, Bole G, Borenstein D, Brandt

K et al (1986) Development of criteria for the classification and reporting of osteoarthritis. Classification of osteoarthritis of the knee. BEZ235 Diagnostic and therapeutic criteria committee of the american rheumatism association. Arthritis Rheum 29:1039–1049CrossRef Altman R, Alarcon G, Appelrouth D, Bloch D, Borenstein D, Brandt K et al (1991) The American college of rheumatology criteria CYT387 manufacturer for the classification

and reporting of osteoarthritis of the hip. Arthritis Rheum 34:505–514CrossRef Berg van den TIJ, Elders LAM, de Zwart BCH, Burdorf A (2009) The effects of work-related and individual factors on the work ability index: a systematic review. Occup Environ Med 66:211–220CrossRef Bieleman HJ, van Ittersum MW, Groothoff JW, Reneman MF, van der Schans CP, Oosterveld FGJ (2007) Arbeidsbelastbaarheid van mensen met beginnende heup- en knieklachten. Een verkennend onderzoek in het CHECK artrosecohort. Work capacity of people with early complaints of hip and knee. An explorative study in the CHECK osteoarthritis cohort. Ned T Fysiotherapie Thiamet G 117(6):225–232 Bieleman HJ, Reneman MF, van Ittersum MW, van der Schans CP, Groothoff JW, Oosterveld FGJ (2009) Self-reported functional status as predictor of observed functional capacity in subjects with early osteoarthritis of the hip and knee. A diagnostic study in the CHECK cohort. J Occup Rehabil 19(4):345–353CrossRef Broersen JP, de Zwart BC, van Dijk FJ, Meijman TF, van Veldhoven M (1996) Health complaints and working conditions experienced in relation to work and age. Occup Environ Med 53(1):51–57CrossRef Brouwer S, Reneman MF, Dijkstra PU, Groothoff JW, Schellekens JM, Goeken LN (2003) Test-retest reliability of the Isernhagen work systems functional capacity evaluation in patients with chronic low back pain.

aureus (198 human isolates and 55 animal isolates) using microarray. Presence or absence of each gene (listed on left) in each isolate is depicted by colour. The colour is an indicator of test signal over reference signal ratio. Thus, (i) yellow indicates presence of the gene in both test strain and reference strain, (ii) red indicates presence of the gene in the test strain but not in the reference strain, (iii) blue indicates absence in the test strain but not the reference strain,

and (iv) grey indicates absence in both the test and reference strains. Genes with white signals are very low intensity and regarded as negative for both strains. The colour intensity is an indicator of signal intensity, and this can differ Foretinib because (i) the homology of the probe, which can be hundreds of base pairs long, and DNA may vary, and (ii) copy numbers may vary. Isolates https://www.selleckchem.com/products/AZD6244.html (represented vertically) are clustered into PD0325901 clinical trial lineages [14]. For each isolate, its mammalian host of origin and its lineage (clonal complex) are shown at the bottom of the figure. Human isolates are coloured light blue (invasive) and dark blue (carriage). Animal isolates are coloured red (cow), pink (horse), maroon (sheep and goat) and white (camel). The figure shows

that rep genes and resistance genes are distributed in a lineage dependent manner. We also assessed the distribution of other plasmid genes between S. aureus lineages. The presence of plasmid conjugation transfer (tra)A-M genes was rare amongst the S. aureus isolates in our collection and was not associated with lineage (Figure 2). Interestingly, antimicrobial resistance genes and heavy metal resistance genes were associated to lineage. arsC was common in MRSA CC22 and CC30 isolates, but rare amongst other lineages.

blaZ was common in all human lineages of S. aureus but was rare in animal lineages of S. aureus. cadA presence was associated with MRSA CC22, CC30 and CC239 lineages, whilst cadDX was widely distributed and associated with 9 different Aprepitant lineages. ermA presence was associated with CC8 and CC239 lineages. qacA was associated with CC239 lineage. 2 of 9 (ble and tetM) resistance genes represented on the microarray are rare in the isolates we have analysed and were not distributed in a lineage dependent manner. We note that some of these genes may be carried on other elements or on integrated plasmids and this cannot be determined by microarray alone, for example tetM can also be carried on transposons such as Tn5801. Discussion In this study we extended a previously proposed plasmid classification system to characterise rep genes from 243 plasmids that appear in the public domain [11]. We characterised 21 rep families, of which 13 are newly described in this study. Whilst performing this analysis we noted that many plasmids carried more than one rep gene, we therefore assigned plasmids into groups based on the combination of rep genes carried.

Moreover, as the sections continued posteriorly, the feeding pocket and the CGS selleck chemicals that surrounded the main rod diminished, and ultimately only the main rod and the accessory rod remained (Figure 6C-D). Serial sections through the anterior region of the nucleus, moving from anterior to posterior, demonstrated the C-shaped curvature of the rod apparatus (Figure 7, 9). These sections also demonstrated how the anterior ends of both the main rod and the accessory rod terminate on the ventral side of the indented nucleus near

the vestibulum (Figure 7F). Similarly, serial sections through the posterior region of the nucleus, moving from anterior to posterior, demonstrated the C-shaped curvature of the rod apparatus and its relationship to the indented nucleus (Figure 8, 9). Flagellar Root System Two flagella emerged from the base of the flagellar pocket (Figure 2A-B, 10A-F, 11A-E). Each VX-770 price flagellum had a paraxial rod (PR) in Palbociclib mouse addition to the 9+2 arrangement of microtubules forming the axoneme (Figure 10G-H, 11F). The PR in the dorsal flagellum (Df) had a whorled disposition, whereas the PR of the ventral flagellum (Vf) had a lattice-like arrangement of parallel fibres (Figure 11F). No mastigonemes were observed on either flagellum (Figure 2A-B). The dorsal basal body contained

a long opaque core (Figure 11B). Both basal bodies were approximately 1.7 μm long and were linked by a connecting fibre (CF) (Figure 10A-B). A cartwheel structure was present at the proximal end of both basal bodies (Figure 10A-B). Two accessory basal bodies (Db’ and Vb’) were observed on the ventral side of the Db and the dorsal side of the Vb (Figure 10B). Figure 10 TEM micrographs showing sections of basal bodies, flagellar roots and associated structures, of Bihospites bacati n.

gen. et sp. A-H from proximal to distal end of flagellar pocket. A-C. Non-consecutive serial sections showing origin and organization of flagellar pocket. A. High magnification TEM of proximal region of basal bodies showing dorsal and ventral basal bodies (Db and Vb) linked by a connecting fibre (CF). Basal bodies with cartwheel structures associated to electron-dense fibres (arrowheads). B. TEM showing accessory dorsal and ventral basal bodies (Db’ and Vb’) on the left of the two main basal bodies. very Dorsal root (DR) connects to electron-dense body (dorsal lamella=DL), on right side of Db. C. TEM showing intermediate root (IR) associated with right side of Vb. Ventral root (VR) associated with electron-dense material that becomes ventral lamella (VL). Row of dorsal microtubules (DMt), not associated with basal bodies. D. Detail of ventral side of Figure C showing Vb, VR formed by four microtubules, VL and intermediate root (arrowhead), initially composed of eight microtubules. E. Detail of dorsal side of Figure C showing DR, with six microtubules (white arrowheads), and DL. F.

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