Cryopreservation of bovine semen in egg yolk based extenders

Cryopreservation of germplasm is widely used in agriculture, biotechnology, conservation of threatened species and human reproductive medicine. There is a need however to improve the reproductive efficiency of breeding with cryopreserved semen, which may involve increasing the post-thaw quality of sperm through improvements in cryopreservation extenders. Extenders including egg yolk from chickens are successfully used worldwide for cryopreservation of bovine semen, whereas the protective agent in the egg yolk is believed to be the low-density lipoprotein (LDL) fraction. Egg yolks of different avian species vary in their cholesterol, phospholipid and polyunsaturated fatty acid content which have been shown to have important effects on sperm’s freezing capability. The purpose of this study was to determine the cryoprotective effect of clarified egg yolk and LDLs extracted from different egg yolk sources (chicken, chicken omega-3, pigeon, quail and turkey) on bovine sperm. Semen from six bulls was collected four times each by electroejaculation, split and diluted with the 10 following extenders: chicken clarified (Ccl), chicken omega-3 clarified (O3cl), pigeon clarified (Pcl), quail clarified (Qcl), turkey clarified (Tcl), chicken LDL (CLDL), chicken omega-3 LDL (O3LDL), pigeon LDL (PLDL), quail LDL (QLDL) and turkey LDL (TLDL). The extended semen was evaluated, cryopreserved and examined directly after thawing (0h) and after two hours at 37 ˚C (2h). Computer assisted sperm analysis (CASA) was used to determine total sperm motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL) and average path velocity (VAP). Intact plasma membrane (IPM) and intact acrosomes (IA) were measured by flow cytometry. The percentage change (loss; Δ%) of each sperm characteristic was calculated and used to compare the effect of the extenders. From extending to 0h post-thaw, the pigeon LDL extender lead to greater losses in sperm total and progressive motility, as well as of intact acrosomes, than the other nine extenders tested (P < 0.05). During 0h to 2h post-thaw, the sperm in PLDL extender experienced greater losses in total and progressive motility (P < 0.0001), as well as in curvilinear velocity (P < 0.05), than in all the other nine extenders. Sperm in turkey clarified extender had a greater loss in the velocity parameters (VSL, VAP, VCL) than sperm in several of the other extenders such as O3cl, CLDL, O3LDL, QLDL and TLDL from 0h to 2h (P < 0.05). Concomitantly, sperm in the Tcl extender had a greater loss in the velocity parameters and of intact acrosomes compared to sperm in its counterpart, the turkey LDL extender, from 0h to 2h post-thaw (P < 0.05). The differences produced in post-thaw quality of cryopreserved bovine sperm in the pigeon LDL and turkey clarified extenders were attributed to methodological differences in these egg yolk preparations compared with the other eight extenders. Importantly, the results demonstrate that with most egg yolk preparations derived from a variety of species, there are equivalent cryoprotective effects afforded by the use of omega-3 chicken, pigeon, quail, or conventional chicken egg yolk in a clarified form in freezing extenders for bovine semen. We further proved that the freezing capabilities of bovine semen extenders containing the low-density lipoprotein fraction of omega-3 chicken, quail, turkey and conventional chicken egg yolk were similar

Physical activity in people with spinal cord injury in Switzerland

Spinal cord injury (SCI) is a devastating health condition by leading to the deterioration or loss of a number of functions depending on an intact spinal cord and consequently to limitations and restrictions in a variety of activities and domains of participation. Due to the decrease of physical capacity, people with SCI have increased risks to develop secondary conditions, which again possibly contribute to premature death. Accordingly, people with SCI still have a shorter life expectancy compared to the general population. Among the leading causes of death in SCI are cardiovascular diseases for which physical inactivity represents a significant risk factor. In general, physical activity (PA) can improve both mental and physical health in people with SCI. People participating in sport or other PAs show lower levels of mental disorders. Furthermore, regular leisure time PA and specific exercises can improve fitness and reduce the risks for secondary conditions. However, to achieve physical health benefits recommendations on PA suggest regular specific endurance and muscle strengthening training with a defined minimum of intensity and duration. In a number of studies in different countries it could be shown that PA levels are ra-ther low, in particular for some subgroups. Since it is likely that PA behavior depends on the culture and context in which people are living, PA levels may vary in different countries. As-sociates and determinants with PA levels have been investigated and identified mainly environmental factors to associate with participation in PAs. However, a comprehensive understanding of the associations with PA rarely exists yet due to the lack of a comprehensive model explaining PA in people with SCI. For the development and implementation of programs to promote PA country specific information is essential to target the programs to the specific needs of the SCI population. For Switzerland, no data on PA levels and according associations with PA levels for people with SCI exist to date. The general objective of this doctoral thesis is to describe PA levels and their associates in people with SCI in Switzerland for the first time. Specifically, it aims to describe LTPA (sport and exercise) levels and the achievement of recommendations on PA in the Swiss SCI population and to identify associations with PA levels by applying comprehensive models. The results should provide stakeholders responsible for the development of programs promoting PA in people with SCI in Switzerland with relevant information. The first study aimed to describe the frequency of participation in sport (PiS) and to identify correlates for PiS in persons with SCI in Switzerland. It could be shown that about 60% of the study sample (n=505) participated in sport at least once a week. However, PiS decreased significantly from the time before the onset of SCI to the time of the survey (P<.001). Sport levels were significantly lower in women than men for the time of the survey (P<.001), whereas no difference was observed before onset of SCI (P=.446). Persons with tetraplegia participated significantly less often in sport than persons with paraplegia (P<.001). Lesion level, active membership in a club, frequency of PiS before the onset of SCI, and the subjective evaluation of the importance of sport correlate with PiS. When controlling for gender differences, only the subjective importance of sport for persons with SCI determines PiS, particularly among women. The objective of the second study was to describe physical activity (PA) levels by considering distinct types and intensities and the achievement of the World Health Organization (WHO) recommendations on PA in persons with SCI and to investigate associated factors. In this study it could be shown that participants (n=485) carried out PA a total of 6.0 hours/week (median) whereas most time was spent to carry out PAs with only light intensity (median 2.2 hours/week). 18.6% were physically inactive, 50.3% carried out muscle-strengthening exercis-es, and 48.9% fulfilled the WHO recommendations. Furthermore, it was found that women, people aged 71+, and people with complete tetraplegia had significantly lower odds of fulfilling the WHO recommendations than participants in the respective reference category (men, ages 17-30, incomplete paraplegia). The third study aimed to explore associations with PA levels by applying a comprehensive model based on the ICF framework. The specific aim was to identify those aspects that potentially explain being physically active and the achievement of the WHO recommendations on PA (ACH-WHO-REC) in people with SCI. This study showed that higher levels of social support and self-efficacy significantly increased the odds of being physically active. For ACH-WHO-REC, the use of an intermittent catheter increased, whereas being dependent in self-care mobility and coping with emotions by focusing on and venting of emotions decreased the respective odds. Furthermore, experiencing hindrances due to accessibility is associated with increased odds for ACH-WHO-REC. For both PA outcomes, older age decreased, but being a manual wheelchair user increased the odds to achieve the outcome

Outcomes of Antiretroviral Therapy in the Swiss HIV Cohort Study: Latent Class Analysis

An in-depth understanding of the different groups that make up the HIV-infected population should inform prevention and care. Using latent class analysis (LCA) we identified seven groups with similar socio-demographic and behavioral characteristics at enrolment in the Swiss HIV Cohort Study: older gay men, younger gay men, older heterosexual men, injection drug users, single migrants, migrant women in partnerships and heterosexual men and women. Outcomes of combination antiretroviral therapy (ART) were analyzed in 1,633 patients starting ART. Compared to older gay men, the probability of a virologic response to ART was reduced in single migrants, in older heterosexual men and in IDUs. Loss to follow-up was higher in single migrants and IDUs, and mortality was increased in older heterosexual men and IDUs. Socio-behavioral groups identified by LCA allow insights above what can be gleaned from traditional transmission groups, and may identify patients who could benefit from targeted intervention

A Mechanogenetic Model of Exercise-Induced Pulmonary Haemorrhage in the Thoroughbred Horse

Exercise-induced pulmonary haemorrhage (EIPH) occurs in horses performing high-intensity athletic activity. The application of physics principles to derive a ‘physical model’, which is coherent with existing physiology and cell biology data, shows that critical parameters for capillary rupture are cell–cell adhesion and cell stiffness (cytoskeleton organisation). Specifically, length of fracture in the capillary is a ratio between the energy involved in cell–cell adhesion and the stiffness of cells suggesting that if the adhesion diminishes and/or that the stiffness of cells increases EIPH is more likely to occur. To identify genes associated with relevant cellular or physiological phenotypes, the physical model was used in a post-genome-wide association study (GWAS) to define gene sets associated with the model parameters. The primary study was a GWAS of EIPH where the phenotype was based on weekly tracheal wash samples collected over a two-year period from 72 horses in a flat race training yard. The EIPH phenotype was determined from cytological analysis of the tracheal wash samples, by scoring for the presence of red blood cells and haemosiderophages. Genotyping was performed using the Illumina Equine SNP50 BeadChip and analysed using linear regression in PLINK. Genes within significant genome regions were selected for sets based on their GeneOntology biological process, and analysed using fastBAT. The gene set analysis showed that genes associated with cell stiffness (cytoskeleton organisation) and blood flow have the most significant impact on EIPH risk

Hepatitis C Virus Infections in the Swiss HIV Cohort Study: A Rapidly Evolving Epidemic

In the Swiss HIV Cohort Study, a nationwide cohort with systematic hepatitis C virus (HCV) infection screening since 1998, HCV incidence decreased in injection drug users, remained low in heterosexuals, and dramatically increased in men who have sex with me

Infrequent Replication of Parvovirus B19 and Erythrovirus Genotypes 2 and 3 among HIV-Infected Patients with Chronic Anemia

We investigated the role that erythroviruses (parvovirus B19 and erythrovirus genotypes 2 and 3) play in the lives of immunosuppressed HIV-infected patients with chronic anemia. We screened the serum samples of 428 patients by specific ultrasensitive real-time polymerase chain reaction assay. Sixteen patients had circulating DNA, with no apparent clinical impact. Erythrovirus-associated anemia is an extremely rare event in HIV-infected patient

Microcephalin and pericentrin regulate mitotic entry via centrosome-associated Chk1

Primary microcephaly, Seckel syndrome, and microcephalic osteodysplastic primordial dwarfism type II (MOPD II) are disorders exhibiting marked microcephaly, with small brain sizes reflecting reduced neuron production during fetal life. Although primary microcephaly can be caused by mutations in microcephalin (MCPH1), cells from patients with Seckel syndrome and MOPD II harbor mutations in ataxia telangiectasia and Rad3 related (ATR) or pericentrin (PCNT), leading to disturbed ATR signaling. In this study, we show that a lack of MCPH1 or PCNT results in a loss of Chk1 from centrosomes with subsequently deregulated activation of centrosomal cyclin B–Cdk1

Tetracationic bis-triarylborane 1, 3-butadiyne as a combined fluorimetric and Raman probe for simultaneous and selective sensing of various DNA, RNA and proteins

A new bis-triarylborane tetracation (4-Ar2B-3, 5-Me2C6H2)-C≡C- C≡C-(3, 5-Me2C6H2-4-BAr2 [Ar = (2, 6-Me2-4-NMe3-C6H2)+] (24+) shows distinctly different behaviour in its fluorimetric response than that of our recently published bis-triarylborane 5- (4-Ar2B-3, 5-Me2C6H2)-2, 2’-(C4H2S)2-5’-(3, 5-Me2C6H2-4-BAr2) (34+). Single-crystal X-ray diffraction data on the neutral bis- triarylborane precursor 2N confirm its rod-like dumbbell structure, which is shown to be important for DNA/RNA targeting and also for BSA protein binding. Fluorimetric titrations with DNA/RNA/BSA revealed the very strong affinity of 24+ and indicated the importance of the properties of the linker connecting the two triarylboranes. Using the butadiyne- rather than a bithiophene linker resulted in an opposite emission effect (quenching vs enhancement), and 24+ bound to BSA 100 times stronger than 34+. Moreover, 24+ interacted strongly with ss-RNA, and circular dichroism (CD) results suggest ss- RNA chain-wrapping around the rod-like bis-triarylborane dumbbell structure like a thread around a spindle, a very unusual mode of binding of ss-RNA with small molecules. Furthermore, 24+ yielded strong Raman/SERS signals, allowing DNA or protein detection at ca. 10 nM concentrations. The above observations, combined with low cytotoxicity, efficient human cell uptake and organelle-selective accumulation make such compounds intriguing novel lead structures for bio-oriented, dual fluorescence/Raman-based applications

Systematic review of clinical practice guidelines for adults with fractures: identification of best evidence for rehabilitation to develop the WHO’s Package of Interventions for Rehabilitation

Background The identification of existing rehabilitation interventions and related evidence represents a crucial step along the development of the World Health Organization’s (WHO) Package of Interventions for Rehabilitation (PIR). The methods for such identification have been developed by the WHO Rehabilitation Programme and Cochrane Rehabilitation under the guidance of the WHO’s Guideline Review Committee secretariat. The aim of this paper is to report on the results of the systematic search for clinical practice guidelines (CPGs) relevant to the rehabilitation of adults with fractures and to present the current state of evidence available from the identified CPGs. Methods This paper is part of the Best Evidence for Rehabilitation (be4rehab) series, developed according to the methodology presented in the World Health Organization’s (WHO) Package of Interventions for Rehabilitation (PIR) introductory paper. It is a systematic review of existing CPGs on fractures in adult population published from 2009 to 2019. Results We identified 23 relevant CPGs after title and abstract screening. According to inclusion/exclusion criteria, we selected 13 CPGs. After checking for quality, publication time, multiprofessionality, and comprehensiveness, we finally included five CPGs dealing with rehabilitative management of fractures in adult population, two CPGs addressing treatment of distal radius fracture and three the treatment of femoral/hip fracture. Conclusion The selected CPGs on management of distal radius and femoral/hip fracture include few recommendations regarding rehabilitation, with overall low to very low quality of evidence and weak/conditional strength of recommendation. Moreover, several gaps in specific rehabilitative topics occur. Further high-quality trials are required to upgrade the quality of the available evidence

Identifying the concepts contained in outcome measures of clinical trials on four internal disorders using the international classification of functioning, disability and health as a reference

Objectives: To systematically identify and compare the concepts contained in outcome measures of clinical trials on chronic ischemic heart disease, diabetes mellitus, obesity and obstructive pulmonary disease, including asthma using the International Classification of Functioning, Disability and Health (ICF) as a reference. Methods: Randomized controlled trials between 1993 and 2003 were located in MEDLINE and selected according predefined criteria. The outcome measures were extracted and the concepts contained in the outcome measures were linked by so called “linking rules” to the ICF. Results: 166 trials on chronic ischemic heart disease, 227 trials on diabetes mellitus, 428 trials on obesity and 253 trials on obstructive pulmonary disease were included. 10 different health status questionnaires were extracted in chronic ischemic heart disease, 19 in diabetes mellitus, 47 in obesity, 39 in obstructive pulmonary disease. Across conditions at least 75% (range: 75-92%) of the extracted concepts could be linked to the ICF. In diabetes mellitus and obesity the most used ICF- categories were ‘general metabolic functions’ (b540), in obstructive pulmonary disease ‘respiratory functions’ (b440) and in chronic ischemic heart disease ‘heart functions’ (b410). Conclusions: In all four health conditions the majority of studies were drug trials focusing on clinically relevant parameters and not on functioning. The ICF provides a useful reference to identify and quantify the concepts contained in outcome measures used in clinical trials