Evaluation of ACE, SP17, and FSHB as candidates for stallion fertility in Hanoverian warmblood horses.

The research of fertility in humans and other mammals has strongly advanced in the recent years. The examination of molecular mechanisms influencing horse fertility is relatively recent. We chose the angiotensin converting enzyme (ACE), the sperm autoantigenic protein 17 (SP17) and the follicle stimulating hormone (FSHB) as candidates for determining stallion fertility and to analyze associations of intragenic single nucleotide polymorphisms (SNPs), flanking microsatellites and candidate-gene linked haplotypes with the pregnancy rate per oestrus (PRO) in 179 Hanoverian stallions. Fertility traits analyzed were the least square means of PRO for stallions (LSMs) and the paternal and embryonic component of breeding values for PRO (BVs). We detected nine SNPs and two flanking microsatellites in ACE, eight SNPs and two flanking microsatellites in SP17 and four SNPs and one flanking microsatellite in FSHB. Three SP17-associated SNPs and the two flanking microsatellites showed significant association with the embryonic component of BVs and one SP17-associated microsatellite was also significantly associated with the paternal component of BVs. Two ACE-associated SNPs were significantly associated with the embryonic component of BVs. Significantly associated haplotypes were shown for all three candidate genes and the tested fertility parameters. The final regression analysis model indicated that haplotypes of all three candidate genes significantly contributed to the paternal and embryonic fertility components of PRO. This is the first report of associations of ACE, SP17 and FSHB with fertility traits of stallions.

Evaluation of prolactin receptor (PRLR) as candidate gene for male fertility in Hanoverian warmblood horses.

Stallion fertility has increasing importance as the artificial insemination is employed in horses more intensely. Molecular genetic markers may be useful tools to evaluate the stallion fertility before breeding. The prolactin receptor gene (PRLR) was chosen as a candidate for stallion fertility because of its influence on testicular and accessory sex gland function. Screening the equine PRLR gene for polymorphisms in Hanoverian stallions revealed two single nucleotide polymorphisms (SNPs). Association and haplotype analyses were performed in 162 Hanoverian warmblood stallions for these intragenic SNPs using the least square means (LSM) of the pregnancy rate per oestrus for stallions and the paternal component and embryonic component of the breeding values (BV) of the pregnancy rate per oestrus. The two SNPs (BIEC2-589441, BIEC2-560860) showed significant associations using single marker and haplotype analysis with the embryonic and paternal component of BV and one SNP (BIEC2-560860) was also significantly associated with the LSM of the pregnancy rate per oestrus. This is the first report on an association of PRLR-associated genetic markers with fertility traits in stallions.

INHBA-associated markers as candidates for stallion fertility.

The inhibin beta A (INHBA) gene was chosen as candidate for stallion fertility and analysed for intragenic markers to find associations with pregnancy rate per oestrus. Intragenic single nucleotide polymorphisms (SNPs) were developed in order to perform an association and haplotype analysis using the least square means (LSM) of the pregnancy rate per oestrus for stallions as well as breeding values (BVs) for the embryonic and paternal component of the pregnancy rate per oestrus. The polymorphisms were genotyped in 161 Hanoverian warmblood stallions. Insemination records from approximately 20,000 Hanoverian warmblood mares were used to calculate LSM for stallions and to predict the paternal and embryonic component of BVs for the pregnancy rate per oestrus. We demonstrated significant associations of single markers and haplotypes with the LSM and the embryonic and paternal component of BVs for the pregnancy rate per oestrus. This is the first report on INHBA as an associated candidate gene with the LSM of stallions and the paternal and embryonic component of BVs for the pregnancy rate per oestrus.

Relaxing music as pre-medication before surgery: a randomised controlled trial.

INTRODUCTION: Patients who await surgery often suffer from fear and anxiety, which can be prevented by anxiolytic drugs. Relaxing music may be an alternative treatment with fewer adverse effects. This randomised clinical trial compared pre-operative midazolam with relaxing music. METHOD: Three hundred and seventy-two patients scheduled for elective surgery were randomised to receive pre-operative prevention of anxiety by 0.05-0.1 mg/kg of midazolam orally or by relaxing music. The main outcome measure was the State Trait Anxiety Inventory (STAI X-1), which was completed by the patients just before and after the intervention. RESULTS: Of the 177 patients who completed the music protocol, the mean and (standard deviation) STAI-state anxiety scores were 34 (8) before and 30 (7) after the intervention. The corresponding scores for the 150 patients in the midazolam group were 36 (8) before and 34 (7) after the intervention. The decline in the STAI-state anxiety score was significantly greater in the music group compared with the midazolam group (P<0.001, 95% confidence interval range -3.8 to -1.8). CONCLUSION: Relaxing music decreases the level of anxiety in a pre-operative setting to a greater extent than orally administrated midazolam. Higher effectiveness and absence of apparent adverse effects makes pre-operative relaxing music a useful alternative to midazolam for pre-medication.

Evaluation of SPATA1-associated markers for stallion fertility.

Stallion fertility is an economically important trait because the use of artificial insemination is increasing in the horse industry and superior sires are used more intensely. Molecular genetic markers may be useful as early indicators for a stallion's fertility and genetic improvement programmes. The testis-specific SPATA1 protein is involved in shaping the sperm head during spermatogenesis. Thus, the spermatogenesis associated 1 (SPATA1) gene was chosen as candidate for stallion fertility, and we analysed intragenic single nucleotide polymorphisms (SNPs) as genetic markers for the least square means (LSM) of the pregnancy rate per oestrus of stallions and breeding values (BV) for the paternal and embryonic component of the pregnancy rate per oestrus. We sequenced the cDNA of SPATA1 to verify the annotated mRNA sequence. One SPATA1-associated intronic SNP (BIEC2-968854) showed a significant association with the embryonic component of BVs of stallions for the pregnancy rate per oestrus. The embryonic component of BVs was positively associated with homozygous C/C stallions. Both the additive and dominance effects were significant with values of -5.8% (P = 0.01) and -6.4% (P = 0.02) for the embryonic component of BVs. For the same SNP, a suggestive association was found for the LSM of the pregnancy rate per oestrus of stallions. Heterozygous stallions had higher pregnancy rates per oestrus than homozygous stallions. The dominance effect was 4.1% with a nominal P-value of 0.02. The SNP BIEC2-968854 can change an SP1 binding site and thus we assume that gene regulation may be influenced through this intronic mutation. This is the first report on SPATA1 being associated with the pregnancy rate per oestrus for stallions.

Postoperative nausea and vomiting after breast surgery: efficacy of prophylactic ondansetron and droperidol in a randomized placebo-controlled study.

Postoperative nausea and vomiting (PONV) is a common adverse phenomenon following breast surgery. The efficacy of ondansetron and droperidol in preventing post-operative nausea and vomiting in women undergoing breast surgery was compared in this randomized, double-blind, placebo-controlled study. Altogether 207 women were randomly assigned to receive either a single intravenous dose of droperidol (1.25 mg) (n = 69), ondansetron (8 mg) (n = 67) or saline (n = 71) immediately after induction of general anaesthesia with thiopental, fentanyl, atracurium, nitrous oxide in oxygen and isoflurane. Complaints of nausea, vomiting and requests for rescue antiemetics were recorded during a 24-h period postoperatively. During the initial 2 h in the postanaesthesia care unit, the incidence of postoperative nausea and vomiting was 15%, 6% and 12% in the placebo, droperidol and ondansetron groups, respectively (NS). The incidence of post-operative nausea and vomiting during the first 24 h was 61%, 48% and 45% in the placebo, droperidol and ondansetron treatment groups, respectively (NS). Postoperative analgesic requirements and the length of stay in the post-anaesthesia care unit were equal in all three treatment groups. It is concluded that the intravenous pretreatment with single doses of ondansetron or droperidol did not substantially prevent postoperative nausea and vomiting after breast surgery.

The modifying influence of anaesthesia on postoperative protein catabolism.

We studied two groups of six patients scheduled for gastrointestinal surgery; they were allocated randomly to receive high- or low-dose fentanyl anaesthesia. The confounding effect of protein balance, before the trauma of surgery, on postoperative nitrogen excretion was controlled by standardized protein intake before operation, supplemented by adequate calories. The high-dose group had significantly lower stress levels during surgery, assessed by arterial blood concentrations of cortisone, adrenaline and glucose. After operation, protein catabolism was measured for 7 days. The high-dose group had significantly lower postoperative excretion of ammonia and slightly lower excretion of urea and 3-methylhistidine. Low-stress anaesthesia may thus diminish postoperative catabolism, which could be important in frail patients by reducing mortality, ICU resources, or both.

Protein and amino acid metabolism during early starvation as reflected by excretion of urea and methylhistidines.

Endogenous excretion of nitrogenous products was studied during early starvation in six healthy, nonobese subjects after six days on a well-defined diet, designed to achieve net protein balance and an adequate calorie supply. The diet contained 0.5 g myofibrillar-free protein and 35 kcal/kg body weight. The subjects then fasted for three days. Urine was collected for 24-hour periods and analyzed for urea, ammonia, 3-methylhistidine, and 1-methylhistidine. Blood glucose and serum urea levels were measured daily. In a second group of subjects, muscle biopsies for determination of free amino acid concentrations were taken in the overnight fasted state and after three days of fasting. During the period with a balanced diet, urea production fell initially and stabilized after two to three days at a level of 146 +/- 15 mmol/24 h. During the period of fasting, serum urea increased from 3.0 +/- 0.4 to a maximum value of 6.2 +/- 0.7 mmol/L and urea production rose markedly, to a peak of 293 +/- 16 mmol/24 h. Ammonia excretion was 24 +/- 2 mmol/24 h before and 71 +/- 13 mmol/24 h after three days of fasting. 3-Methylhistidine excretion was stable before fasting and then rose from 154 +/- 17 to 198 +/- 17 mumol/24 h. 1-Methylhistidine excretion was unchanged during fasting. Blood glucose levels were stable at 4.8 +/- 0.2 mmol/L before fasting and then fell to 3.7 +/- 0.3 mmol/L. Intracellular concentrations of amino acids in skeletal muscle decreased markedly during fasting; after three days of fasting the glutamine concentration had fallen by 34%.(ABSTRACT TRUNCATED AT 250 WORDS)

Paravertebral block during cholecystectomy: effects on circulatory and hormonal responses.

Surgical trauma induces a hormonal metabolic response which is partly responsible for postoperative catabolism. In this study 12 patients underwent cholecystectomy during isoflurane anaesthesia, six with a paravertebral block (PVB) in addition. Plasma concentrations of glucose, cortisol and adrenaline, and heart rate and arterial pressure were compared between the two groups. The patients with PVB showed a significantly diminished response to noxious stimuli.

High- and low-dose fentanyl anaesthesia: hormonal and metabolic responses during cholecystectomy.

We have compared two groups of patients given low- or high-dose fentanyl anaesthesia. Arterial blood samples were collected for measurement of glucose, free fatty acids (FFA), glycerol, beta-hydroxy-butyrate, insulin, c-peptide, glucagon, human growth hormone (HGH), cortisol and adrenaline concentrations. After induction of anaesthesia, blood concentrations of most of these substances decreased. After the start of surgery the concentrations of cortisol, glucose, HGH, FFA and beta-hydroxy-butyrate increased significantly in the group anaesthetized with the lower dose of fentanyl. In the group that received high-dose fentanyl anaesthesia the plasma concentrations of almost all the hormones and substances measured remained relatively low. The differences between the two groups during surgery were significant for adrenaline (P less than 0.001) and cortisol (P less than 0.001). High-dose fentanyl appears to block the trauma-induced stress response seen in patients anaesthetized with low dose fentanyl.

High- and low-dose fentanyl anaesthesia: circulatory and plasma catecholamine responses during cholecystectomy.

The cardiovascular and neuroendocrine effects of a high-dose fentanyl anaesthesia (100 micrograms/kg body weight) were compared with those of a balanced type of fentanyl anaesthesia (5 micrograms kg-1) during upper abdominal surgery. High-dose fentanyl anaesthesia prevented the increase in catecholamine concentrations and attenuated the circulatory response to surgical stress seen in the group anaesthetized with the balanced technique of anaesthesia.

Multiple mRNAs are generated from the chicken lysozyme gene.

We have determined the DNA sequence of a 770 bp Pst 1 fragment containing 450 nucleotides of the 5' flanking region of the chicken lysozyme gene. S1-nuclease mapping was performed to localize the 5' end of nuclear RNA containing lysozyme-specific sequences and of the mRNA. We present evidence that the 5' noncoding region of the chicken lysozyme mRNA is heterogeneous in length. The 5' termini of the different nRNAs map 29, 31 and 53 nucleotides upstream from their common initiation codon. The 5' ends of lysozyme-specific nuclear RNAs map at positions similar to that of the mRNA. AT-rich regions and sequences similar to the E. coli RNA polymerase recognition sequence are found around 30 and 70 nucleotides upstream from each of these 5' termini. The AT-rich regions differ, however, from the canonical Goldberg-Hogness box in that they do not contain the extremely conserved TATA sequence motif. Sequence comparison at the 5' end of the lysozyme, conalbumin and ovalbumin genes reveals only one region of partial homology, 140 nucleotides upstream from the mRNA start sites.

[Organization of genes (author's transl)]. / Organisation von Genen.

The organization of genes of animal cells is discussed. The genes for 5S-RNA, rRNA and for histones are examples of the organization of repetitive genes. The interruptions of the coding sequence of the mouse beta-globin gene illustrates the split nature of many eukaryotic genes. It is shown that changes in the organization of genes do occur during the normal development of an organism. The methods used for the analysis of gene structure are of great importance for elucidation of the molecular basis of genetic diseases like the thalassemias.

Chicken lysozyme gene contains several intervening sequences.

The organization of the chicken lysozyme gene and its neighboring sequences was examined by a comparison of the restriction map of the lysozyme structural gene with the map of the lysozyme gene in genomic DNA. Chicken DNA was cleaved with restriction endonucleases and the DNA fragments were separated by agarose gel electrophoresis. After transfer of the fragments onto nitrocellulose filters, those fragments that contain lysozyme mRNA sequences were detected by hybridization of the filters to labeled probes generated from pls-1, a recombinant plasmid carrying the lysozyme structural gene. This analysis revealed the presence of at least three intervening sequences, two of which interrupt the protein coding region and one of which is located in the 3' untranslated region. When oviduct DNA and sperm DNA were compared, no difference was observed in the size and number of restriction fragments that contain either lysozyme or ovalbumin structural gene sequences.

Transcription of the chicken ovalbumin and conalbumin gene during early secondary induction with estrogens.

Estrogens stimulate the rate of transcription of the ovalbumin and conalbumin gene in the chicken oviduct. The synthesis of ovalbumin and conalbumin mRNA was studied in isolated nuclei. RNA synthesized in vitro was distinguished from preexisting nuclear RNA by affinity labeling the in vitro products with a mercurated nucleotide and subsequent purification of the Hg-RNA on SH-agarose. The content of ovalbumin and conalbumin mRNA sequences in the in vitro transcripts was determined by hybridization to cDNA. After the withdrawal of implanted hormones from chickens, the synthesis of conalbumin and ovalbumin RNA increased 2.5- and at least 20-fold, respectively, by treatment with estrogens. The maximal rate of transcription of the conalbumin gene is achieved within 2 h after estrogen induction, whereas the rate of transcription of the ovalbumin gene becomes maximal after a lag of several hours. These results demonstrate that estrogens affect two genes in the same target cell differently.