Sub-chronic (13-week) oral toxicity study in rats with recombinant human lactoferrin produced in the milk of transgenic cows.

The oral toxicity of recombinant human lactoferrin (rhLF) produced in the milk of transgenic cows was investigated in Wistar rats by daily administration via oral gavage for 13 consecutive weeks, 7 days per week. The study used four groups of 20 rats/sex/dose. The control group received physiological saline and the three test groups received daily doses of 200, 600 and 2000 mg of rhLF per kg body weight. Clinical observations, growth, food consumption, food conversion efficiency, water consumption, neurobehavioural testing, ophthalmoscopy, haematology, clinical chemistry, renal concentration test, urinalysis, organ weights and gross examination at necropsy and microscopic examination of various organs and tissues were used as criteria for detecting the effects of treatment. Overall, no treatment-related, toxicologically significant changes were observed. The few findings that may be related to the treatment (lower cholesterol in high-dose females, lower urinary pH in high-dose males and females and very slightly higher kidney weight in high-dose females) were considered of no toxicological significance. Based on the absence of treatment-related, toxicologically relevant changes, the no-observed-adverse-effect level (NOAEL) was considered to be at least 2000 mg/kg body weight/day.

Postnatal health and welfare of offspring conceived in vitro: a case for epidemiological studies.

In vitro fertilization (IVF) has established itself as an important technique in human assisted reproduction and in livestock improvement. In both humans and livestock the possible long-term effects on health and welfare of offspring born after IVF and in vitro culture to the blastocyst stage are still largely unknown. Epidemiological studies in humans, using data collected for individuals born after normal (i.e. non-assisted) pregnancies, have provided evidence for associations between prenatal life events and adult-life disease. Due to the relatively short time that elapsed since the first IVF baby was born, comparable studies for IVF offspring are not yet possible. However, animal experiments and epidemiological studies with the available data from the livestock industry (mainly dairy cattle) may contribute to a better understanding of the risks involved.

Evaluation of three brands of tympanic thermometer.

Infrared tympanic thermometers (ITT) have many documented benefits, including speed, ease of use, and noninvasiveness, to support their use in emergency departments (ED) and intensive care units (ICU). However, concerns have been raised about the accuracy of temperatures reported by ITT. This study was conducted to evaluate the accuracy of 3 brands of ITT, compared to rectal and pulmonary artery thermometers, in ED and ICU settings. Results indicated adequate specificity for all 3 ITT in ED (range = 0.9242 to 1.0000) and ICU (range = 0.9737 to 1.0000), but unacceptable sensitivity in ED (range = 0.5455 to 0.8000) and ICU (range = 0.0000). Further analyses indicated highly variable ITT-reported temperatures. In ED, ITT temperatures were, on average, 0.3684 degree C lower, and could be expected to vary by more than 2 degrees C from the actual temperature as reported by rectal thermometer. In ICU, the average reported temperature was similar to the actual pulmonary artery temperature (0.0259 degree C lower), but again could vary by more than 2 degrees C. Recommendations for nursing and education are discussed.

"Specific" scale compared with "generic" scale: a double measurement of the quality of life in a French community sample of obese subjects.

STUDY OBJECTIVE: To determine the aspects of quality of life that are affected by weight in the general population, to develop a specific questionnaire (OSQOL) that can measure with reliability and validity the impact of overweight and obesity on well being, and to compare the results with those obtained using a well known generic tool (the "SF 36" scale). DESIGN: Cross sectional survey with matched control group. SETTING: Community. PATIENTS: A permanent survey base of approximately 10,000 representative ordinary households were screened for weight and height. Five hundred subjects were randomly drawn from the sub-sample of adult people with a body mass index equal or greater than 27. A control sample of 500 subjects matched for sex, age, and employment status was drawn from the non-obese population. MAIN RESULTS: A short specific Quality Of Life scale, the OSQOL, was produced that comprises 11 items belonging to four independent dimensions. Using this scale and the SF36, it was found that: (1) moderately obese subjects (27 < or = BMI < 30) did not significantly differ from the control group except for physical capacity; (2) in the group of obese subjects with a BMI > 30, quality of life seemed to be impaired for five of nine dimensions of the SF36 compared with the control population, all related to physical consequences of obesity. This population essentially perceived itself in terms of poor general health. (3) No significant difference was observed between the samples for the psychological and social dimensions of the SF36. CONCLUSION: The quality of life of patients with severe obesity is impaired, but it mainly affects the physical consequences of the disease. The psychological and social repercussions that could have been expected to affect this population were not demonstrated. The hypothesis of a process of adaptation of the person and their social environment cannot therefore be excluded. Methodologically, comparison of the specific OSQOL with the generic SF3 shows clearly that the two kinds of scales correspond to different objectives and should be considered as complementary.

Converting a presentation to a manuscript.

Presenting and publishing are closely linked. Although many authors are later asked to speak, the reverse is also true. Many presenters are asked to author manuscripts based on their presentations. This author who is experienced in professional presentations and editing gives tips to nurse authors on how to develop a manuscript from a presentation.

Follicular wall maintains meiotic arrest in bovine oocytes cultured in vitro.

Cumulus oocyte complexes (COCs) were cocultured with parts of the follicular wall. Coculture conditions were such that the COCs were 1) in continuous contact with the follicular wall (FWC), 2) separated from the follicular wall at collection but in contact with it during culture (FWR), and 3) separated from the follicular wall, but cultured in its vicinity (FWNR). Oocytes cultured for 24 hr under FWC conditions maintained the germinal vesicle stage. Under FWR conditions the germinal vesicle stage was not maintained, but an arrest at metaphase I of meiosis occurred in most of the oocytes. When COCs were cultured in the vicinity of the follicular wall (FWNR), meiosis was resumed and similar numbers of oocytes progressed to metaphase II of meiosis as compared to cultures of COCs without coculture with parts of the follicular wall. When COCs were isolated from the follicular wall after 24 hr of culture and additionally cultured for another 24 hr, the oocytes showed the same capability of resuming meiosis as fresh, isolated cumulus oocyte complexes. It is concluded that maintenance of contact with the follicular wall is necessary to maintain meiotic arrest. When COCs restore a physical contact with the follicular wall during culture, an arrest at metaphase I occurs.

When industry courts your clinical research skills, should you collaborate?

Hospital/industry collaboration is as yet rare in Canadian nursing research. Dupont Canada Inc. invited this research team to become one of four sites taking part in a study of decubitus ulcers in intensive care unit patients. The project was regarded as an opportunity to study an important nursing problem. Collaboration involved a multicenter meeting to plan the research with personnel from the four sites and data collection on the incidence of pressure sores in intensive care unit patients. Industry contributed funding for the project, work on the overall research design, coordination of the sites, and data analysis. The hospitals contributed clinical knowledge, access to the patients, and skilled nurses to collect skin care data. The authors have gained from the collaboration, as has industry. In this article, we review the process, problems, and benefits of collaboration with industry and provide recommendations for nurses getting involved in hospital/industry collaborative research.

Bovine activin A does not affect the in vitro maturation of bovine oocytes.

The effect of recombinant bovine activin A on the in vitro maturation of bovine oocytes was investigated. Culture of cumulus enclosed bovine oocytes in the presence of activin at the concentration of 100 or 500 ng/ml did not change the proportion of oocytes in which germinal vesicle breakdown had occurred at 4 and 7 h after the onset of culture. Activin had also no effect on the progression of maturation to the M II stage. The transient inhibition of germinal vesicle breakdown by 10 mM dibutyryl cyclic AMP was not affected by the addition of activin A at the onset of culture. Radiolabeling with 35S-methionine at 4 h and at 18 h after culture in the presence or absence of activin A did not show any effect of activin either on the total incorporation of radiolabel into acid precipitable material or on the protein synthesis patterns obtained after SDS-PAGE.

Evaluation of transvaginal ultrasound-guided follicle puncture to collect oocytes and follicular fluids at consecutive times relative to the preovulatory LH surge in eCG/PG-treated cows.

Holstein-Friesian cows (n=56) were synchronized with Syncro-Mate B, and those cows (n=47) developing a normal progesterone pattern were further treated im with 3,000 I.U. eCG at Day 10 and 22.5 mg PGF2alpha 48 h later. Blood samples were collected every hour from 30 until 49 h after PG administration. Cows (n=17, 36.2%) with fewer than 8 follicles larger than 8 mm in diameter at 28 to 30 h after PG treatment and animals without an LH peak (n=7, 23%) were excluded from the study. Transvaginal ultrasound-guided puncture of the follicles was carried out two times per cow, at 30 h after PG injection (4 to 5 follicles) and again at 1 to 5 (n=6), 12 (n=8) or 22 h (n=9) after the LH peak. No differences in the concentrations of progesterone and LH were observed among the 3 groups. An average of 18 follicles per cow was punctured (total of 415 punctures, n=23); 116 cumulus-oocyte-complexes and 370 follicular fluid samples were obtained producing average recovery rates of 28.0% and 89.2%. The number of cumulus-oocyte-complexes varied between puncture times; shortly before ovulation, at 22 h after the LH peak, the recovery rate was significantly 5 times higher than immediately after the LH peak. Overall, in 75 punctures the cumulus-oocyte-complex was accompanied by a pure follicular fluid sample (3.3 per cow). In conclusion, the transvaginal ultrasound-guided puncture of preovulatory-size follicles can be used to collect follicular fluids to study changes in the microenvironment of maturing oocytes upon superovulation. However, further research is required in order to obtain an equivalent number of accompanying cumulus-oocyte-complexes.

Stage-specific appearance of the mouse antigen TEC-3 in normal and nuclear transfer bovine embryos: re-expression after nuclear transfer.

Bovine embryos, recovered from the uterus in vivo or derived from in vitro matured and in vitro fertilized oocytes, were investigated for the presence of the developmentally regulated mouse antigen TEC-3 by indirect immunofluorescence. During preimplantation embryo development TEC-3 is expressed on bovine morulae and blastocysts. It is absent from unfertilized and fertilized oocytes, and from all stages before the 32-cell stage. The finding that TEC-3 is not expressed before the onset of embryonic transcription, which occurs at the eight-cell stage in the bovine, but only when the embryonic genome is active, makes it a potential marker for studying nuclear reprogramming after nuclear transfer. Nuclear transfer embryos were made by electrical fusion of blastomeres from morulae derived in vivo with enucleated metaphase II oocytes. Indirect immunofluorescence with the TEC-03 antibody showed that the TEC-3 antigen, present on blastomeres of the morula stage embryo, disappeared after fusion and was expressed again when the nuclear transfer embryos developed to the morula and blastocyst stage. These data suggest that the bovine embryonic nucleus may be able to revert to the equivalent of an earlier developmental stage when transferred to ooplasm, and is then capable of following the normal developmental program.

Development of a sperm zona pellucida binding assay for bull semen.

Assays based on sperm zona pellucida binding have been developed as diagnostic tests to predict the fertilisation potential of human spermatozoa. The aim of this study was to establish a similar assay for bull sperm. The results showed differences between established fertile bulls in the relative numbers of sperm cells bound to the zona pellucida of a batch of oocytes. These differences suggest that there may be a relationship between the sperm zona pellucida binding capacity and the fertility of bulls.

Understanding mediastinitis: nursing's role in prevention and treatment.

Mediastinitis is a clinical problem seen in patients after a mediasternotomy usually following cardiac surgery. Although relatively rare, this complication slows recovery, lengthens hospital stay and increases health care costs. An overview of the problem of sternal wound infection after cardiac surgery is provided. Predisposing, intraoperative and post-operative factors related to this problem will be discussed. Attention to the factors related to the problem may contribute to the prevention of mediastinitis. Clinical recognition of the problem will be described and treatment options, nursing therapeutics and research questions will be addressed.

Structural aspects of bovine oocyte maturation in vitro.

Bovine cumulus-oocyte complexes (COCs) were collected from 4-8 mm follicles and graded into four categories on their morphological characteristics. These four categories were matured in vitro and processed for transmission electron microscopy at 24 h after the onset of culture. The morphology of the four groups of oocytes was analysed and compared with that of oocytes that had matured in vivo and were collected 20-23 h after the preovulatory luteinizing hormone peak. After in vivo maturation, oocytes formed a homogeneous group with respect to their morphological characteristics. After in vitro maturation, the oocytes formed a heterogeneous group with respect to their morphology between as well as within the four categories of oocytes. Oocytes from the first three categories showed the same morphology after in vitro maturation. The fourth category showed some specific characteristics: 1) vacuolization, 2) flattening of cumulus cells, and 3) almost complete lack of cortical granules in some category 4 oocytes. These characteristics are interpreted as signs of degeneration. Besides these aspects of degeneration, other deviations from normal development were seen: 1) retraction of cumulus cell process endings from the oocyte without the breaking down of these processes, 2) retardation of some aspects of the cytoplasmic maturation, and 3) incomplete cumulus expansion. It is concluded that oocytes capable of development in vitro show a large morphological variability before the onset of culture. In vitro maturation systems can support normal development, but many oocytes show signs of degeneration and deviant development after in vitro maturation.

Heterologous cell contacts and metabolic coupling in bovine cumulus oocyte complexes.

The integrity of the cumulus cell processes were studied in four categories of bovine cumulus oocyte complexes (COCs) selected on their morphological characteristics. Three different types of cumulus cell process endings (CCPEs) were identified, one penetrating the cortex, another not penetrating the cortex, and a third form was intermediate and more rare in appearance. The process endings that penetrated the cortex frequently made gap junctions with the oolemma. The division of the three types of CCPEs over the four different COC categories was specific for three of the four categories. The first-category COC predominantly possessed the penetrating CCPE, the fourth-category COC possessed predominantly the nonpenetrating CCPE, and the second and third categories had both types of CCPEs. The metabolic coupling of the cumulus-oocyte contacts was assessed by means of incorporation of 3H-choline into the oocyte. The majority of category 4 COCs transferred low levels of choline into the oocyte while the majority of the oocytes of the other three categories transferred high levels of choline into the oocyte. Category 4 includes a smaller proportion of oocytes capable of cleaving after fertilization than the other three categories. This reduced developmental capacity is probably due to the loss of metabolic coupling before the onset of culture.

Morphology of preovulatory bovine follicles as related to oocyte maturation.

Thirty-three preovulatory bovine oocytes and their follicles were collected during the period of final maturation in normally cyclic cows. Cell density of the membrana granulosa, mitotic index of the membrana granulosa, and the occurrence of eosinophilic granulocytes around the basal membrane as well as the maturational stage of the oocyte were determined. Cell density decreased during the period of final maturation. Mitotic indices also decreased after an initial high level in the first hours of the final maturation. Eosinophilic granulocytes were only seen during the last hours of final maturation. The maturational stages of the oocytes were related to distinct maturational stages of the follicular wall as determined by morphological characteristics. We propose a scoring system for the maturity of the follicular wall based on cell density, presence of mitotic figures and the presence of eosinophilic granulocytes outside the vascular compartment.

Follicular and oocyte maturation in cows treated for superovulation.

The maturational stage of oocytes and their follicles was assessed at 24/26 h after the preovulatory luteinizing hormone (LH) peak by means of morphological criteria. Follicles were obtained from cows treated for superovulation (PMSG/PG) with additional anti-PMSG to neutralize the residual PMSG. Follicular fluid was also recovered and analyzed for progesterone and estradiol levels. Seventy-two percent of the oocytes were at the Metaphase II (M(II)) stage of meiosis, whereas only 28% of the follicular walls were at the proper maturational stage; assessed on morphological characteristics, 78% of the follicles were progesterone-dominated. Earlier maturational stages of oocytes and follicles were also present, including those that are restricted to periods shortly after the LH peak in the normally cyclic cow. It is concluded that upon treatment for superovulation not all oocytes and follicles mature synchronously, and that not all oocytes mature in harmony with their follicles.

Circular questions: a family interviewing strategy.

Assessment strategies for working with the family of the critically ill typically focus on collecting data from family members in order to determine if the family requires further support or intervention. This article describes using the technique of circular questioning to obtain information from and provide the family with new information. Circular questions are interventions as well as assessment strategies. The authors provide critical care examples of the use of circular questions with families of the critically ill.