bluuebunny/crossref_metadata_2025 huggingface dataset:
Litigation related to inadequate anaesthesia: an analysis of claims against the NHS in England 1995–2007
Authors: Mihai R. , Scott S. , Cook T. M.
Published: July 2009

Inadequate anaesthesia may cause distress to the patient and lead to medical litigation. All claims made to the NHS Litigation Authority 1995–2007 were obtained and the data was examined independently by all authors and classified. In a dataset of 1067 claims there were 161 cases of inadequate anaesthesia and data were suitable for analysis in 159: intra‐operative awareness (79), brief awake paralysis (20) and inadequate regional anaesthesia (60). The total cost of closed claims was £3.2m. Cost was incurred in 100% of claims of brief awake paralysis, 87% of claims of awareness and 80% of claims of inadequate regional blockade. Mean cost of closed claims was £32 680 for anaesthetic awareness, £29 345 for inadequate regional blockade and £24 364 for brief awake paralysis. Inadequate anaesthesia accounts for 19% of anaesthesia‐related claims in the NHS in England. Strategies that reduce anaesthetic awareness, drug errors and inadequate regional blockade are known and their improved implementation is likely to reduce such claims.
Monitor position and the accuracy and speed of ultrasound‐guided nerve blocks
Authors: Langford R. A. , Hockey B. , Leslie K.
Published: July 2009

Ultrasound is rapidly becoming an essential skill for all anaesthetists with an interest in regional anaesthesia. Using a single‐blinded cross‐over trial design, we assessed the accuracy and speed of simulated ultrasound‐guided nerve blockade with the monitor either in direct line‐of‐sight or perpendicular to the line‐of‐sight. Thirty‐one anaesthetists participated in the study. Times taken to reach the targets were similar 18.4 (14.6) s with the monitor ‘in‐line’ and 22.4 (26.7) s with the monitor perpendicular to the target. Needle placement was significantly more accurate when anaesthetists had the monitor in front of them (29 acceptable) than to the side (22 acceptable). For anaesthetists with limited ultrasound experience, the accuracy, but not speed, of ultrasound guided regional anaesthesia can be improved by aligning the monitor in the line of sight of the operator.
The impact of manual in‐line stabilisation on ventilation and visualisation of the glottis with the LMA CTrach™: a randomised crossover trial*
Authors: Ng B. S. W. , Goy R. W. L. , Bain J. A. , Chen F. G. , Liu E. H. C.
Published: July 2009

The LMA CTrach™ (CTrach) enables ventilation, glottis visualisation and tracheal intubation via a laryngeal mask conduit. The CTrach has been successfully used in patients with cervical spine pathology, but it is unclear if cervical spine immobilisation affects its ease of use. In this randomised crossover trial, the CTrach was used once with and once without manual in‐line stabilisation of the cervical spine in every patient. With manual in‐line stabilisation, the median [IQR] time to achieve ventilation was 22 [16–32] s, compared with 19 [13–30] s without stabilisation (p = 0.065). With manual in‐line stabilisation, the time to achieving a glottic views was 42 [30–63] s compared with 39 [25–53] s without stabilisation (p = 0.019). There was no difference in the success rates of achieving ventilation and glottic views. These results suggest that manual in‐line stabilisation does not affect use of the CTrach.
Myoclonus after cardiac arrest: pitfalls in diagnosis and prognosis*
Authors: English W. A. , Giffin N. J. , Nolan J. P.
Published: July 2009

Accurate prediction of neurological outcome in survivors of cardiac arrest may be difficult. We report the case of a 44‐year‐old survivor of a hypoxic cardiac arrest who repeatedly developed relentless myoclonic jerks on attempted discontinuation of his propofol infusion. These were initially thought to represent myoclonic status epilepticus before the correct diagnosis of Lance–Adams syndrome was made. Lance–Adams syndrome is a rare disorder seen in survivors of profound hypoxic episodes. It is characterised by intention myoclonus but preserved intellect. Accurate distinction between myoclonic status epilepticus and Lance–Adams syndrome is vital as they have very different prognoses. The different pathophysiology and distinguishing clinical features of these two conditions are highlighted.
Peri‐operative oral triiodothyronine replacement therapy to prevent postoperative low triiodothyronine state following valvular heart surgery
Authors: Choi Y. S. , Kwak Y. L. , Kim J. C. , Chun D. H. , Hong S. W. , Shim J. K.
Published: July 2009

This study evaluated the effect of oral triiodothyronine (T3) replacement therapy, starting on the day of the surgery, on thyroid hormone concentrations and clinical outcome in high‐risk patients undergoing valvular heart surgery. Fifty patients were randomly allocated to either T3 or placebo. In the treatment (T3) group patients received 20 μg of oral or nasogastric T3 every 12 h starting just before induction of anaesthesia and until the first day after surgery. T3 concentrations were significantly higher in the T3 group than the placebo group from 1 to 36 h after removal of the aortic cross clamp. The number of patients requiring vasopressin after discontinuing cardiopulmonary bypass was significantly greater in the placebo group than the T3 group. Significantly fewer patients required vasopressors in the T3 group on the first day after surgery.
Void detection in root fillings using intraoral analogue, intraoral digital and cone beam CT images
Authors: Huybrechts B. , Bud M. , Bergmans L. , Lambrechts P. , Jacobs R.
Published: July 2009

Aim  To compare void detection in root fillings using different radiographic imaging techniques: intraoral analogue, intraoral digital and cone beam CT (CBCT) images and to assess factors influencing small void detection.

Methodology  Two straight root canals in canine teeth were prepared. Calibrated steel wires of five different diameters (200, 300, 350, 500, 800 μm) were inserted respectively in the canal after the injection of a sealer. To simulate filling voids of known dimensions, the wires were removed after the sealer had set. Each sample was imaged, using a Minray X‐ray tube (Soredex, Helsinki, Finland) at optimal clinical settings combined with Vistascan PSP (Dürr Dental, Bietigheim‐Bissingen, Germany), Digora Optime PSP (Soredex), Sigma CCD (Instrumentarium, Tuusula, Finland) and E‐speed films (Agfa‐Gevaert, Mortsel, Belgium). The teeth were also imaged using CBCT (3D Accuitomo, Morita, Japan). A generalized mixed model and anova analysis were used on the acquired data (Tukey–Kramer correction).

Results  There was no evidence that the factor ‘root level’ affected void detection in root fillings. ‘Void size’ was a main determining factor as all voids larger than 300 μm were determined with all techniques. For the smaller voids, there were significant differences between the 5 imaging techniques at different void sizes and different root levels.

Conclusions  Void size and imaging technique were main determining factors. Voids larger than 300 μm were determined with all imaging techniques. For small void detection, all digital intraoral techniques performed better than intraoral analogue and CBCT images.
Mycobacterial PE, PPE and ESX clusters: novel insights into the secretion of these most unusual protein families
Authors: Bottai Daria , Brosch Roland
Published: July 2009

The human pathogen Mycobacterium tuberculosis harbours a large number of genes that encode proteins whose N‐termini contain the characteristic motifs Pro–Glu (PE) or Pro–Pro–Glu (PPE). A subgroup of the PE proteins contains polymorphic GC‐rich sequences (PGRS), while a subgroup of the PPE proteins contains major polymorphic tandem repeats (MPTR). The function of most of these proteins remains unknown. However, in this issue of Molecular Microbiology, Abdallah and colleagues show that PE_PGRS proteins from the model organism Mycobacterium marinum are secreted by components of the ESX‐5 system that belongs to the recently defined type VII secretion systems. These observations, which now need to be addressed and confirmed in M. tuberculosis, open new perspectives on the function of these highly abundant proteins.
Direct stimulus perception and transcription activation by a membrane‐bound DNA binding protein
Authors: Gebhard Susanne , Gaballa Ahmed , Helmann John D. , Cook Gregory M.
Published: July 2009

Few membrane proteins with a role in transcriptional regulation have been studied, and none are able to perceive their respective stimuli and activate transcription of their regulons without the aid of auxiliary proteins. The bacitracin resistance regulator, BcrR, of Enterococcus faecalis is a membrane‐bound DNA binding protein and is required for bacitracin‐dependent expression of the bacitracin resistance genes, bcrABD. Here, we show that BcrR interacts directly with Zn2+ bacitracin (Kd = 2–5 μM), but not metal‐free bacitracin. A solution‐based DNA binding assay demonstrated that the affinity of BcrR for its target DNA is much higher (Kd = 40 nM) than previously found for transmembrane regulators and is comparable to that of soluble DNA binding proteins. A construct of BcrR that lacked the transmembrane domain was unable to bind to DNA, indicating that membrane localization was important for DNA binding. Bacitracin did not cause a change in the DNaseI footprint of BcrR on the bcrA promoter, but in vitro transcription assays with BcrR proteoliposomes showed bacitracin‐dependent activation of transcription. These findings demonstrate that BcrR is a bona fide one‐component transmembrane signal transduction system, which perceives an extracellular stimulus (presence of bacitracin) and relays it to an intracellular transcriptional response independent of any auxiliary proteins.
Characterization of starvation‐induced dispersion in Pseudomonas putida biofilms: genetic elements and molecular mechanisms
Authors: Gjermansen Morten , Nilsson Martin , Yang Liang , Tolker‐Nielsen Tim
Published: July 2009

Pseudomonas putida OUS82 biofilm dispersal was previously shown to be dependent on the gene PP0164 (here designated lapG). Sequence and structural analysis has suggested that the LapG geneproduct belongs to a family of cysteine proteinases that function in the modification of bacterial surface proteins. We provide evidence that LapG is involved in P. putida OUS82 biofilm dispersal through modification of the outer membrane‐associated protein LapA. While the P. putida lapG mutant formed more biofilm than the wild‐type, P. putida lapA and P. putida lapAG mutants displayed decreased surface adhesion and were deficient in subsequent biofilm formation, suggesting that LapG affects LapA, and that the LapA protein functions both as a surface adhesin and as a biofilm matrix component. Lowering of the intracellular c‐di‐GMP level via induction of an EAL domain protein led to dispersal of P. putida wild‐type biofilm but did not disperse P. putida lapG biofilm, indicating that LapG exerts its activity on LapA in response to a decrease in the intracellular c‐di‐GMP level. In addition, evidence is provided that associated to LapA a cellulase‐degradable exopolysaccharide is part of the P. putida biofilm matrix.
Dehydroepiandrosterone sulphate improves cholestasis‐associated fatigue in bile duct ligated rats
Authors: Butterworth R. F. , Lalonde R. , Power C. , Baker G. B. , Gamrani H. , Ahboucha S.
Published: July 2009

Abstract:  Fatigue is a common debilitating symptom in patients with primary biliary cirrhosis (PBC). The mechanism of fatigue is still poorly understood. However, it has been reported that levels of the steroid dehydroepiandrosterone sulphate (DHEAS) are reduced in plasma of patients with PBC, and substitutive therapy has been suggested to improve fatigue symptoms experienced during the course of this disease. In this study, we tested the effect of DHEAS on whole body fatigue in rats following bile duct ligation (BDL). Fatigue was estimated by the time spent on an electrified grid as a result of falling off a treadmill and by performance of rats on an infrared beam monitor which allows the assessment of travelled distance and stereotypic movement activities. On day 5 after BDL surgery, cholestatic rats exhibited increased whole body fatigue as reflected by significantly increased time spent on the electrified grid, reduced travelled distance and reduced stereotypic movements. Administration of 5 mg kg−1 of DHEAS to BDL rats for three consecutive days significantly normalized their behaviour. Fatigue scores were also found to be reduced in cirrhotic rats 4 weeks after BDL surgery, and DHEAS treatment for 3 days reduced fatigue scores at this stage. Dehydroepiandrosterone sulphate treatment was sufficient to increase brain levels of DHEAS in the BDL rats in a manner that is significantly and highly correlated with those of plasma DHEAS and brain dehydroepiandrosterone (DHEA). Substitutive therapies with DHEAS or DHEA could represent novel approaches in the management of fatigue due to cholestasis‐induced liver failure.
Nuclear and cytoplasmic maturation of canine oocytes related to in vitro denudation
Authors: Chebrout M , De Lesegno C Viaris , Reynaud K , Chat S , Chastant‐Maillard S
Published: July 2009

Contents

In vitro maturation (IVM) of bitch oocytes is, to date, a very inefficient process, with common metaphase rates approximately 0–20% and mean degeneration rates approximately 20–30%. In other mammals, meiotic resumption is controlled in the cumulus–oocyte complex by the disappearance of the coupling between granulosa cells and the oocyte. The first aim of this study was to evaluate the influence on meiotic resumption of a mechanical denudation of the oocytes before maturation. The nuclear stage was determined by DNA staining with ethidium‐homodimer‐2 under confocal microscopy. Denuded (n = 318) and control (n = 378; no mechanical denudation) oocytes had similar degeneration rates (respectively 32.1 vs 28.6%). However, meiosis resumption rates were significantly higher for denuded oocytes (DO; 26.9 vs 17.8%). Secondly, we aimed to evaluate oocytes experiencing spontaneous denudation during the 72 h IVM period. Denuded oocytes, having lost cumulus cells on at least 75% of their perimeter (n = 440), were compared with surrounded oocytes (SO), with 100% of their perimeter surrounded by granulosa cells (n = 860). As above, the nuclear stage was determined by confocal microscopy, but cytoplasmic maturation was also evaluated through transmission electron microscopy. Degeneration rates but also meiosis resumption and metaphase rates were significantly higher for denuded than for SO (9.6 vs 2.8% for metaphase rate). Nevertheless, ultrastructurally, metaphase DO have scarcer organelles unevenly distributed, with smooth endoplasmic reticulum concentrated in aggregates in the cortical zone. Denudation, whether mechanical or spontaneous, is thus an inefficient mean to obtain metaphase II oocytes suitable for in vitro fertilization.
Non Cat‐Like Ovarian Cycle in the Eurasian and the Iberian Lynx – Ultrasonographical and Endocrinological Analysis
Authors: Göritz F , Dehnhard M , Hildebrandt TB , Naidenko SV , Vargas A , Martinez F , López‐Bao JV , Palomares F , Jewgenow K
Published: July 2009

Contents

The Iberian lynx is considered the most endangered felid species. Therefore, anex situconservation program was initiated to protect this species from extinction. Additional knowledge on lynx reproduction biology and reliable methods for reproductive monitoring are important for developing a captive breeding program. The aim of this study in lynx was to implement transrectal ultrasonography to visualize ovarian structures (follicles, corpora lutea) and to assess ovarian activity in addition to analysis of serum progesterone and oestradiol. Because of limited access to Iberian lynxes, the less‐endangered Eurasian lynx and bobcat were also studied in this comparative study. Recent endocrinological studies based on faecal and urinary progesterone and oestrogen metabolites revealed that steroid profiles in both these species were alike and did not follow the typical pattern of other felids. Pregnancy diagnosis was not possible, since progesterone concentrations did not differ between pregnant and pseudopregnant animals. Progesterone was also detected after parturition as well as after weaning until the onset of a new oestrous cycle. In the present study, the presence of corpora lutea during the non‐breeding season was confirmed by ultrasonography and by elevated serum levels of progesterone averaging 3.56 ± 1.3 ng/ml in Eurasian and 6.1 ± 0.26 ng/ml in Iberian lynx, respectively. The ultrasonographical findings on the ovarian structures suggest strongly that corpora lutea developed after ovulation stay active until November and regress before the onset of the next oestrus.
Seasonal Profiles of Ovarian Activity in Iberian lynx (Lynx pardinus) Based on Urinary Hormone Metabolite Analyses
Authors: Jewgenow K , Göritz F , Vargas A , Dehnhard M
Published: July 2009

Contents

The Iberian Lynx Ex‐Situ Conservation Programme is an essential part of a co‐ordinated action plan to conserve the most endangered felid species of the world. Successful captive breeding demands reliable methods for reproduction monitoring including reliable non‐invasive pregnancy diagnosis. During a 3‐year study, urine samples from six captive Iberian lynx females were obtained (one non‐pregnant, one pseudo‐pregnant and 11 pregnant cycles). Progesterone, pregnanediol and oestradiol were determined in urinary extracts and relevant urinary oestrogen metabolites were characterized by high‐performance liquid chromatography (HPLC). Urinary progestins did not follow the typical pregnancy‐related course of felids. In the lynx, we failed to demonstrate an urinary progestin elevation during pregnancy. In contrast, urinary oestrogens increased from 3.8 ± 0.6 to 8.6 ± 0.5 ng/mg creatinine (p < 0.001) during the pregnancy. A comparison of pseudo‐pregnant with pregnant cycles revealed a further increase of oestrogens caused by implantation (p < 0.05). In one female, which refused to mate, no difference was estimated between oestrogens levels during the breeding and non‐breeding seasons. Almost 10‐fold higher oestrogen concentrations were measured in urines of females that shared enclosures with males. HPLC analysis of oestrogens in urine samples collected from Iberian lynx during the pregnancy revealed that lynx urine is composed of two polar oestrogen metabolites in addition to oestrone and minor amounts of oestradiol. Oestrone was detectable in all urinary extracts (8–12% of metabolites), whereas oestradiol was elevated only during late pregnancy (18%). Thus, seasonal luteal activity in Iberian lynx can be monitored by urinary oestrogens. The increase of urinary oestradiol during late pregnancy might indicate an oestradiol secretion by the lynx placenta.
Comparison Between Vestibular and Subcutaneous Insertion of Deslorelin Implants for Oestrus Induction in Bitches
Authors: Kutzler M , Lamb SV , Volkmann D
Published: July 2009

Contents

Investigations using sustained‐release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1–4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations >100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.
Prostaglandin Synthesis Genes are Differentially Transcripted in Normal and Pyometra Endometria of Bitches
Authors: Silva E , Leitão S , Ferreira‐Dias G , Lopes da Costa L , Mateus L
Published: July 2009

Contents

Pro‐inflammatory stimuli, such as endotoxins released by Gram‐negative bacteria, are potent stimulators of prostaglandin (PG) synthesis. The aim of this study was to evaluate the gene transcription pattern of PG synthesis enzymes in normal (anestrous, n = 6 and diestrous, n = 8) and pyometra (n = 7) endometria of bitches. Uteri were collected during routine ovariohysterectomy, processed for histopathological evaluation and uterine contents cultured. Gene transcription of COX‐1, COX‐2, mPGES‐1 and PGF‐synthase (PGFS) were evaluated by relative real‐time PCR and normalized with the ribosomal protein L27 (RPL27) housekeeping gene. Normal uteri had no histological abnormalities and were negative for bacteriology. All pyometra uteri were hyperplasic and Escherichia coli was the only isolated bacterium. Except for COX‐1, gene transcription was significantly higher in pyometra than in normal endometria. No significant differences in gene transcription were observed between normal diestrous and anestrous endometria. COX‐2 gene transcription was 19 and 69 times higher in pyometra than in diestrous and anestrous endometria (p < 0.001), while PGFS gene transcription had a 3‐ and 600‐fold increase in pyometra endometria compared to normal diestrous and anestrous endometria (p < 0.001). Gene transcription of mPGES‐1 was 9 times higher in pyometra than in normal uteri (p < 0.01). Based on these results, we suggest that pyometra‐associated E. coli endotoxin release stimulates the up‐regulation of COX‐2 PGFS and mPGES‐1 gene transcription in the endometrium.
Expression Pattern of Markers in the Canine Ovarian Cycle
Authors: Thuróczy J , Oppe N , Scheuermann E , Perge E , Tibold A , Balogh L
Published: July 2009

Contents

Although hormonal changes during different phases of the oestrous cycle of bitches are well‐described, knowledge about the luteal phase and anoestrus is incomplete. Furthermore, which paracrine and autocrine critical factors that differentiate between follicles destined for atresia and those that continue to develop are unknown. In this study, ovarian tissue was collected from 39 healthy bitches that were subject to ovariectomy or ovariohysterectomy for surgical neutering or medical purposes such as unwanted pregnancy. Bitches were allocated to different groups depending on the stage of the oestrous cycle. Serum progesterone, LH, FSH and 17β‐estradiol (E2) ‐levels were determined and immunhistochemistry was performed for a variety of receptor antigens; Ki‐67, vimentin, pan cytokeratin antibody, p53 and oestrogen receptor (ER) α antigens. Marked differences were found in progesterone concentration between pregnant and non‐pregnant animals. Oestrogen concentration was significantly lower in pro‐oestrus and ovulation than during the luteal phase. Although progesterone could be detected in cytoplasm of ovarian cells at each stage, its presence was restricted to follicular cells during anoestrus. A strong presence of AE1/AE3, vimentin and p53 was found in each oestrous stage, in contrast with Ki67. The localization of ERα appeared to vary during the oestrous cycle, a phenomenon that suggests a switch between target cells of oestrogen; while as a proliferation marker, the mild reaction of p53 during parturition suggests an apoptotic process at this stage of the cycle.
Progesterone, Oestradiol, FSH and LH Concentrations in Serum of Progesterone‐Treated Pregnant Bitches with Suspected Luteal Insufficiency
Authors: Tibold A , Thuróczy J
Published: July 2009

Contents

In the bitch, the corpus luteum is the only source of circulating progesterone throughout pregnancy. Inadequate luteal function may be a cause of abortion or foetal resorption observed after early pregnancy diagnosis. In our study of factors involved in canine luteal inadequacy, 35 pregnant bitches from different breeds were allocated to groups of healthy control (n = 15) vs hypoluteoid (n = 20) pregnant bitches, based on presence or absence of clinical signs of impending abortion and on progesterone concentrations below 10 ng/ml at the 4th–5th week of pregnancy. Hypoluteoid bitches were treated daily with 10‐mg natural progesterone in injectable form (Luteosan® inj.; Alvetra and Werfft AG, Vienna, Austria) until day 60. Serum progesterone, 17β‐oestradiol, FSH and LH concentrations were measured in samples obtained weekly using ELISA previously validated for dog serum. The exogenous progesterone supplementation was presumed to be sufficient to prevent foetal loss in 15 of the 20 treated bitches. The mean serum progesterone concentration in control pregnant bitches did not decrease below 10 ng/ml until the 8th week of pregnancy. Progesterone concentrations in progesterone‐treated hypoluteoid bitches at week 4 were lower than in controls (p < 0.05), but although numerically higher did not differ significantly from those of controls during the period of treatment. Serum 17β‐oestradiol concentrations of healthy bitches were variable, were at most time higher than those of treated animals and slowly decreased until parturition; those of treated bitches remained unchanged during the study. Serum FSH and LH concentrations did not differ between groups. Additional studies involving untreated pregnancies showing equivalent evidence of hypoluteoidism as well as assay of circulating relaxin and prolactin in treated and untreated bitches are needed to better determine the causes and effects of hypoluteoidism in pregnant bitches.
Comparison of Two Doses of the GnRH Antagonist, Acyline, for Pregnancy Termination in Bitches
Authors: Valiente C , Corrada Y , De La Sota PE , Blanco PG , Arias D , Gobello C
Published: July 2009

Contents

Gonadotropin‐releasing hormone (GnRH) antagonists are particularly useful when a rapid inhibitory effect on the gonadal axis is required. The aim of this study was to test the efficacy and clinical safety of a low and high dose of the third generation GnRH antagonist, acyline, on pregnancy termination in female dogs. The effect of the antagonist on the progesterone (P4) serum concentration was also described. Twenty‐one mid‐pregnant bitches were randomly assigned to a single subcutaneous (SC) dose of a placebo (PLACE; n = 7), a low (ACY‐L; 110 μg/kg; n = 6) or high (ACY‐H; 330 μg/kg; n = 8) dose of acyline. The animals were followed up for 15 days. All ACY treated but no placebo‐treated animals terminated their pregnancy by abortion (p < 0.01). The ACY‐L and ACY‐H groups interrupted their pregnancy 7 ± 1.9 and 6.4 ± 1.3 days after treatment, respectively (p = 0.7). A significant interaction between treatment and day was found (p < 0.01) for P4 serum concentrations when PLACE was compared with both ACY groups. No difference was found for this hormone between both ACY groups (p > 0.05) where P4 diminished throughout the study. The decreasing rate varied among animals and was closely related to the time of abortion when P4 reached basal concentrations. In PLACE animals, gestation progressed normally and P4 did not change throughout the study (p > 0.05). None of the bitches presented side effects. It was concluded that acyline safely terminated mid‐pregnancy by permanently decreasing P4 serum concentrations.
Dystocia in Numbers – Evidence‐Based Parameters for Intervention in the Dog: Causes for Dystocia and Treatment Recommendations*
Authors: Münnich Andrea , Küchenmeister Uwe
Published: July 2009

Contents

Dystocia is a frequent problem in the dog, with regard to breed‐specific factors, litter size, duration of expulsion, various intervals between puppies and various causes of maternal, foetal or combined origin. In the present study, results were obtained from management of 530 dogs of 54 breeds, between 1 and 13 years of age, presented with dystocia over a period of 8 years in an obstetrical clinic with mostly identical diagnostic and therapeutic schedules. Data on age of the dam, parturition number, breed size, causes of dystocia, duration of expulsion time, kind of obstetrical aid, and last but not least, the influence of all therapeutic aspects on survival of the puppies were analysed. Dams of miniature and small breeds (59.4%) had a high incidence for dystocia in this population. Uterine forces (inertia and spasm), malpresentation of the foetus and litter size (single pup pregnancies, hyperfoetation) were the most common causes for dystocia. Duration of expulsion stage had the highest influence on puppy survival (p < 0.001). In the case of medical treatment, medication with oxytocin led to higher puppy losses compared with other medicaments. Older primiparous bitches (>6 years of age) had a significantly higher risk to have special obstetric conditions and stillbirths compared with young primiparous bitches. The scope of this study was to analyse the parameters that were most useful for a rapid diagnosis to maximize puppy survival, to avoid false diagnosis and to choose the best therapeutic strategy. The present data show that in addition to the established criteria for management of dystocia in the dog, new diagnostic approaches are necessary to improve obstetrical care.
Preliminary Study in Immature Canine Oocytes Stained with Brilliant Cresyl Blue and Obtained From Bitches with Low and High Progesterone Serum Profiles
Authors: Rodrigues BA , Rodriguez P , Silva AEF , Cavalcante LF , Feltrin C , Rodrigues JL
Published: July 2009

Contents

This study was conducted: (i) to observe the features and levels of blue colour impregnation in morphologically selected immature canine cumulus oocyte complexes (COCs) stained with the brilliant cresyl blue (BCB) dye, as indicators of quality, and integrity of nuclear oocyte chromatin configuration before in vitro maturation (IVM); (ii) to observe the relationship between the influence of serum progesterone (SP) concentrations from ovary donors and BCB staining of immature dog oocytes. The results showed that out of 138 canine COCs, germinal vesicle (GV) stage prevailed in BCB+ oocytes at percentages of 67.4% (60/89), which were statistically higher than those observed in BCB+/− (52.2%; 23/44) and BCB− (20%; 1/5) oocytes (p = 0.023). Oocytes BCB+ were interpreted as those having completed their growth and therefore possessing the capacity to mature and develop in vitro. Ooplasm and cumulus cells (CCs) of canine oocytes were BCB staining independent. Ooplasm blue colour staining reaction varied between grown oocytes, revealing different levels of glucose‐6‐phosphate dehydrogenase activity among and within oocytes. Additionally, SP profile of ovary donors was not a relevant indicator for selection of oocytes screened with the BCB stain. Similar numbers of high quality oocytes were observed to be BCB+, BCB+/− and BCB− between groups of females with SP varying from 0 to 2.5 ng/ml (n = 5), and those with SP varying from 2.6 to 16.7 ng/ml (n = 4) (p = 0.680). It may be inferred that bitches with low and high SP profiles have grown oocytes in their ovaries, as determined by the BCB absorbance in their ooplasms.
Isolation of Canine Mammary Cells With Stem Cell Properties and Tumour‐Initiating Potential
Authors: Cocola C , Anastasi P , Astigiano S , Piscitelli E , Pelucchi P , Vilardo L , Bertoli G , Beccaglia M , Veronesi MC , Sanzone S , Barbieri O. , Reinbold RA , Luvoni GC , Zucchi I
Published: July 2009

Contents

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self‐renewal or by progenitor cells that have acquired the capacity for cell self‐renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long‐term non‐adherent culture conditions. We showed that cells obtained from spheres that display self‐renewing properties, have multi‐lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour‐initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.
Oestrogen and Progesterone Receptor Gene Expression in Canine Oocytes and Cumulus Cells Throughout the Oestrous Cycle
Authors: Gonçalves JSA , Vannucchi CI , Braga FC , Paula‐Lopes FF , Milazzotto MP , Assumpção MEOA , Visintin JA
Published: July 2009

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The aim of this research was to analyze oestrogen receptor‐α (ERα), ERβ and progesterone receptor (PR) gene expression in the canine oocyte and cumulus cells throughout the oestrous cycle. Ovaries from 38 bitches were recovered after ovariohysterectomy and sliced. The phase of the oestrous cycle was determined by vaginal cytology, vaginoscopy and serum hormonal measurements. Oocytes were mechanically denuded by repeated pipetting. For each phase of the cycle, a sample was composed by a pool of 50 oocytes (sample number: prooestrus = 3, oestrus = 8, dioestrus = 5 and anoestrus = 5) or a pool of cumulus cells (prooestrus = 4, oestrus = 7, dioestrus = 4 and anoestrus = 6). Oocyte and cumulus cells’ total RNA was isolated and reverse transcription was conducted to perform real‐time PCR. Oestrogen receptor‐α was expressed throughout the cycle in the oocyte (33.33%, 25.0%, 20.0% and 60.0% for prooestrus, oestrus, dioestrus and anoestrus, respectively) and cumulus cells (50.0%, 47.14%, 25.0% and 66.67% for prooestrus, oestrus, dioestrus and anoestrus, respectively). In the oocyte, the ERβ was also expressed in all phases of the cycle (33.33%, 50.0%, 20.0% and 60.0% for prooestrus, oestrus, dioestrus and anoestrus, respectively), whereas in cumulus cells, ERβ was only expressed during prooestrus (50%) and oestrus (14.29%). Interestingly, while the oocyte PR was not detected in any phase of the cycle, this receptor was expressed during prooestrus (50%), oestrus (42.86%) and anoestrus (16.67%) in cumulus cells. In conclusion, canine oocytes express ERα and ERβ throughout the oestrous cycle, however, there is a lack of PR expression in all these phases. Moreover, in cumulus cells, only ERα was expressed throughout the oestrous cycle.
Expression of the μ Opioid Receptor and Effects of the Opioid Antagonist Naloxone onIn VitroMaturation of Oocytes Recovered from Anoestrous Bitches
Authors: Iorga AI , Valentini L , De Santis T , Ambruosi B , Albrizio M , Guaricci AC , Caira M , Dell’Aquila ME
Published: July 2009

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The μ‐opioid receptor (MOR) is expressed in bovine, human, equine and canine oocytes, and in seasonal breeders, it is expressed with higher intensity during the anoestrous phase. Supplementation ofin vitromaturation (IVM) medium with opioid agents, agonists or antagonists, was shown to affect oocyte maturation in several species such as rat, bovine and equine. This study reports the effects of supplementing IVM medium with naloxone (Nx), an opioid antagonist, on nuclear and cytoplasmic maturation rate of oocytes recovered from anoestrous bitches. Cytoplasmic maturation was examined in terms of mitochondrial (mt) distribution. In order to confirm the receptor‐mediated action of Nx, in oocytes of anoestrous bitches, MOR expression was analyzed by Western blot. Cumulus–oocyte complexes, recovered from the ovaries of bitches in anoestrous, were culturedin vitroand Nx was added at the concentrations of 1 × 10−6, 1 × 10−8and 1 × 10−10 m. The rate of oocytes resuming meiosis after culture in presence of 1 × 10−6 mNx (29%) was significantly higher than that of oocytes of control group (12%; p < 0.05). However, treatment with Nx did not affect mt distribution pattern. In denuded oocytes and in corresponding cumulus cells, a doublet of 65 and 50 kDa was observed. We conclude that, in oocytes of anoestrous bitches, MOR is expressed and Nx significantly improves nuclear maturation rate. Further studies should be performed to elucidate the expression of other opioid receptors, such as δ and κ, and possible interactive effects of their antagonists on canine oocyte maturation.
Effects of Short‐term Hyper‐ and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and ‐Thyroid Axis and on Semen Quality in Male Beagles
Authors: Koivisto MB , Eschricht F , Urhausen C , Hoppen HO , Beyerbach M , Oei CHY , Günzel‐Apel AR
Published: July 2009

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Effects of a short‐term hyper‐ and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3‐day intervals for 12 weeks. The time span was divided into five 3‐week periods: pre‐treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 μg/kg orally once daily), post‐treatment 1 and post‐treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15‐week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin‐releasing hormone (TRH; 10 μg/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid‐stimulating hormone and thyroxine (T4) were investigated. Pre‐treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight‐line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH‐related changes. Basic T4 levels were significantly reduced after CAB treatment (p < 0.05). The results of the present study demonstrate that MCP‐induced short‐term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo‐pituitary axis and semen quality.
Pregnancy Monitoring in Dogs and Cats Using 3D and 4D Ultrasonography
Authors: Hildebrandt TB , Drews B , Kurz J , Hermes R , Yang S , Göritz F
Published: July 2009

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Three‐dimensional (3D)/four‐dimensional (4D) volume ultrasound is an established method in human medicine that offers various options for analysing and presenting ultrasound volume data. However, the successful application of the different 3D/4D imaging modalities in pregnant dogs and cats has not yet been reported in the literature. The main reasons for this are: (1) the high costs of 3D/4D ultrasound systems, (2) operation difficulties due to high breathing frequency in non‐sedated animals and (3) the missing specific knowledge in veterinary medicine concerning how to perform high‐quality volume scans. Automatically acquired ultrasound volume data sets were generated with two different ultrasound systems: the portable Voluson i and the stationary Voluson Expert 730. Different 3D/4D imaging modalities were tested in regard of their practicability in pregnancy monitoring in dogs and cats. Nine different volume imaging modalities were applied using the saved files. For the presentation of the static 3D volume data sets, we used the multiplanar, niche, surface, transparency, glass body, inversion, volume calculation and tomographic ultrasound imaging modes. For the dynamic 4D data, the surface and glass body modes were applied. By changing the human standard settings to the requirements of small animal anatomy, it was found that 3D/4D ultrasound has great potential for the characterization of pregnancy in queens and bitches. The 3D/4D technology offered advanced information about pregnancy status and birth prediction and improved the diagnostic confidence. By using standardized examination protocols, 3D/4D ultrasound will allow a reduction in examination time by generating even more relevant information. These benefits, combined with possible future cost reduction of commercial ultrasound systems, might lead to frequent utilization in routine pregnancy diagnostic and birth management in small animal practice.
Impact of 24‐h Cooling Prior to Freezing on the Survival of Domestic Cat (Felis catus) Epididymal Sperm
Authors: Martins JLA , Villaverde AISB , Lima AFM , Steagall PVM , Ferreira JCP , Taconeli CA , Lopes MD
Published: July 2009

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The aim of this study was to investigate the impact of a 24‐h cooling period prior to freezing on domestic cat epididymal sperm viability. Fifteen tomcats were submitted to routine orchiectomy and sperm samples were retrieved from both epididymides in a Tris–glucose–20% egg yolk extender. For each tomcat, the diluted sperm was split into two equal volumes and cooled to 5°C at a rate of 0.5°C/min; one sample for 60 min (control) and the other for 24 h (cooled). After the cooling period, samples from both groups were frozen using an identical freezing protocol. Sperm samples were evaluated in three different periods: immediately after harvesting, after cooling at 5°C for 24 h (cooled group) and after freezing–thawing of control and cooled groups. Evaluations consisted of sperm motility and progressive status, sperm morphology and plasma membrane integrity (PMI) using two fluorescent probes. After cooling for 24 h, a decrease (p < 0.05) in sperm motility, progressive status and PMI was observed when compared to sperm samples immediately after collection. Comparing the results obtained after thawing, no difference (p < 0.05) was found regarding sperm motility, progressive status, PMI and sperm morphology between control and cooled groups. The results from the present study show that cooling cat epididymal spermatozoa at 5°C for 24 h prior to freezing does not lead to major damage of spermatozoa impairing the freeze–thaw process.
Influence of Ascorbic Acid and Glutathione Antioxidants on Frozen‐Thawed Canine Semen
Authors: Monteiro JC , Gonçalves JSA , Rodrigues JA , Lúcio CF , Silva LCG , Assumpção MEOA , Vannucchi CI
Published: July 2009

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Poor sperm viability post‐thaw has resulted in constant research into methods of cryopreservation of canine semen. One factor that may be involved in poor viability is sperm oxidative stress caused by excessive formation of reactive oxygen species. The present study was performed in order to evaluate the effect of different concentrations of ascorbic acid (AA) and glutathione (Glu) added to an extender for the freeze‐thawing of dog sperm. Semen from five mature dogs was collected and frozen in two studies. Prior to and after freezing, sperm motility, morphology and membrane status were examined. In addition, sperm motility was examined up to 120 min after thawing to evaluate thermo‐resistance. In study I, semen was collected twice from each dog. On both occasions, semen was divided into three aliquots: control, Glu 1 mm and Glu 5 mm. In study II, semen was collected twice and divided into three aliquots; control, AA 50 μm and AA 250 μm. Initial sperm motility was significantly higher in sperm diluted with AA 50 μm; sperm longevity, however, measured by a thermal‐resistance test (TRT), was higher for Glu treatments. Higher concentration of Glu produced significant improvement in TRT and membrane status, whereas higher concentration of AA had a negative impact in sperm longevity. Antioxidant supplementation to semen freezing extenders improved semen quality post‐thaw. Moreover, Glu had the most beneficial effect when supplemented at 5 mm.
Effects of the Inclusion of Equex STM into Tris‐Based Extender on the Motility of Dog Spermatozoa Incubated at 5°C
Authors: Niżański W , Klimowicz M , Partyka A , Savić M , Dubiel A
Published: July 2009

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The aim of this study was to estimate the effects of Equex STM on sperm motion characteristics in chilled dog semen extended in Tris‐based diluent. Thirty‐two ejaculates were collected from 12 proven German shepherd stud dogs. The sperm‐rich fractions were diluted in Tris‐based extender with 1% (v/v) Equex STM (sample A) and in Tris‐based extender with no addition of detergent (sample B). The extended semen was incubated for 240 h at 5°C and the motility parameters were evaluated by CASA system at 24‐h intervals. Addition of Equex STM to Tris‐based extender led to an initial activation of motion activity of spermatozoa, followed by a rapid decrease, shortening the lifespan of spermatozoa incubated at 5°C. Computer‐assisted sperm analysis clearly showed that Equex STM‐induced changes of sperm motion characteristics resemble the hyperactivation (HA) of spermatozoa associated with the capacitation process.
Clinical Use of Testicular Fine Needle Aspiration Cytology in Oligozoospermic and Azoospermic Dogs
Authors: Romagnoli S , Bonaccini P , Stelletta C , Garolla A , Menegazzo M , Foresta C , Mollo A , Milani C , Gelli D
Published: July 2009

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Clinical investigation of canine testicular function is complicated by the difficulty in the evaluation of seminiferous tubules. Until recently, testicular biopsy was the only diagnostic option for dogs with persistent oligo/azoospermia. In human andrology, testicular fine needle aspiration (TFNA) is currently considered a useful method in the evaluation of azoospermia and severe oligozoospermia, and has long replaced classical biopsy to evaluate spermatogenesis. In order to verify its diagnostic efficacy for the clinical approach to canine oligo‐ or azoospermia, TFNA was performed in seven adult (two oligozoospermic and five azoospermic) dogs. After sedation, a fine (21–23 gauge) butterfly needle connected to a 50‐ml syringe was inserted into each testicle; strong suction was applied and the aspirated fluid squirted on a glass slide, smeared out, air‐dried and stained with a modified May–Grunwald–Giemsa. Under light microscopy, Sertoli cells (all those found in each investigated field) and spermatogenic cells (n = 100) were counted on each smear in order to differentiate spermatogonia, primary spermatocytes, secondary spermatocytes, early spermatids, late spermatids and spermatozoa, and calculate their relative percentages. Cytological analysis showed the following testicular pictures: normal spermatogenesis (compatible with obstruction of the seminal ducts), hypospermatogenesis, maturative disturbances and Sertoli cell‐only syndrome. Two dogs with an obstructive lesion were treated with corticosteroids; one of them recovered and sired two litters of puppies.