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At least 37 records · Page 2Linked to original sources

Trans-shell infection by pathogenic micro-organisms reduces the shelf life of non-incubated bird's eggs: a constraint on the onset of incubation?

Many birds initiate incubation before clutch completion, which results in asynchronous hatching. The ensuing within-brood size disparity often places later-hatched nestlings at a developmental disadvantage, but the functional significance of the timing of the onset of incubation is poorly understood. Early incubation may serve to maintain the viability of early-laid eggs, which declines over time owing to the putative effects of ambient temperature. An unexplored risk to egg viability is trans-shell infection by micro-organisms. We experimentally investigated the rate and magnitude of microbial trans-shell infection of the egg, and the relative effects of ambient temperature and micro-organisms on hatching success. We show that infection of egg contents is prevalent and occurs within the time required to lay a clutch. The probability of infection depends on the climatic conditions, the exposure period and the phylogenetic composition of the eggshell microbiota. We also demonstrate that microbial infection and ambient temperature act independently to reduce egg viability considerably. Our results suggest that these two factors could affect the onset of avian incubation in a wide range of environments.

Analysis of Variance↗

Production of a healthy calf by somatic cell nuclear transfer without micromanipulators and carbon dioxide incubators using the Handmade Cloning (HMC) and the Submarine Incubation System (SIS).

The aim of this work was to investigate the minimum technical requirements for production of live offspring with somatic cell nuclear transfer. The experiment was performed in a field type laboratory without micromanipulators and carbon dioxide incubators. All long-term incubations were performed in the Submarine Incubation System (SIS) using various gas mixtures. The somatic cell culture was established from ear biopsy of a 9-year-old Holstein cow. Nuclear transfer was performed using the Handmade Cloning (HMC) technique. Zona-free oocytes were randomly bisected by hand with a disposable blade and a stereomicroscope. Cytoplast were selected using Hoechst staining and a fluorescent microscope. After a two-step fusion embryos were activated with calcium ionophore and dimethylaminopurine. Embryos were cultured in microwells (WOWs) in SOFaaci medium supplemented with 5% cattle serum. In two consecutive experiments, six blastocysts were produced from 52 reconstructed embryos. On Day 7, five blastocysts were transferred into synchronized recipients. All three recipients became pregnant but two pregnancies aborted at 6 and 7 months, respectively. A heifer calf weighing 27 kg was delivered at term by Caesarean section from the third pregnancy. The healthy 6-month-old heifer, the first cloned animal of Africa, is living evidence that nuclear transfer technology may be successfully used under basic laboratory conditions.

Animals↗

Cardiovascular responses to thoracic skin cooling: comparison of incubating and non-incubating Bantam hens.

Body temperatures, metabolic rate, haemostatic parameters, and cardiovascular reactions to thoracic skin cooling were compared between incubating (broody) and non-broody Bantam hens. Under resting conditions, without thoracic skin cooling, cardiac output of broody hens was twice that of non-broody hens. However, their metabolic rate (VO2) was increased by only one-third over that of non-broody hens, and the arteriovenous difference in oxygen concentration was smaller for broody birds. This indicates a higher rate of non-nutrient blood flow during incubation. A higher thoracic skin temperature (Tths) for broody hens compared to non-broody hens suggests that brood patches are the probable site of this increased flow through arteriovenous anastomoses (AVAs). Thoracic skin cooling increased metabolic rate and Q significantly more in broody hens, but did not increase AVA blood flow. The relation between metabolic rate and total peripheral resistance indicated more intense vasodilation for broody hens at the relatively low metabolic rates during moderate cooling, and more intense vasoconstriction for the broody hens at the high metabolic rates during stronger cooling. This corresponds to Tths measurements indicating dilation of brood patch AVAs with moderate cooling and AVA constriction with severe cooling. During moderate cooling, vasoconstriction in the feet and wattles of broody hens (but not of non-broody hens) freed non-nutrient blood flow for redistribution to the brood patches. Thus, the cardiovascular system of the hen seems to adjust to the special demands of incubation by a permanent increase of AVA flow in the brood patch, and by an additional capacity for brood patch vasodilation induced by cold stimuli in the range from 35 to 25 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Primary trabecular meshwork cells incubated in human aqueous humor differ from cells incubated in serum supplements.

PURPOSE: To determine whether aqueous humor, the in vivo source of nutrients for trabecular meshwork cells, alters cellular and molecular characteristics in primary trabecular monolayer cell cultures when compared with standard culture conditions. METHODS: Human primary trabecular meshwork cell cultures were grown in DMEM supplemented with 50% human aqueous humor (DMEM-AH), heat-denatured DMEM-AH, 10% fetal bovine serum (DMEM-FBS, the standard culture supplement), or heat-denatured DMEM-FBS. Confluent trabecular cells were assayed for cell propagation and morphology for 21 days. Protein expression profiles of trabecular cell lysates were analyzed by two-dimensional polyacrylamide gel electrophoresis. Western blot analysis was used to determine the protein expression of myocilin and TIMP-1 in conditioned media collected from trabecular cells at 5, 10, 15, and 21 days. Myocilin expression was also analyzed by Western immunoblots after addition of dexamethasone (10(-7) M) or ascorbic acid (29 mg/dL). RESULTS: Trabecular cells supplemented with DMEM-AH for 21 days showed decreased cell proliferation when compared with DMEM-FBS (11% vs. 141%). Cellular morphology was also altered: Trabecular cells incubated in DMEM-AH showed larger-, broader-, and flatter-appearing cells than did the more spindle-shaped cells grown in DMEM-FBS. Protein profiles of trabecular cell lysates isolated from cells incubated in DMEM-AH differed from those incubated in DMEM-FBS. In DMEM-AH-conditioned medium, myocilin expression was increased and TIMP-1 expression was decreased at day 21. Induction of myocilin by dexamethasone was observed in conditioned medium isolated from cells treated with DMEM-FBS (442%), but only a 10% increase in myocilin was observed beyond the normal induction in DMEM-AH. Daily administration of ascorbic acid to DMEM-AH failed to increase myocilin expression beyond that obtained with DMEM-AH. CONCLUSIONS: Addition of human aqueous humor rather than FBS to trabecular monolayer cell cultures triggers significant changes in cellular and molecular characteristics. The protein component of aqueous humor is responsible for these changes. Aqueous humor supplementation may maintain cultured trabecular cells in a more physiologic state.

Adult↗

Prevalence of detectable abnormal prion protein in persons incubating vCJD: plausible incubation periods and cautious inference.

BACKGROUND: Both small and large variant Creutzfeldt Jakob disease (vCJD) epidemics are consistent with the current observed incidence. Uncertainty in vCJD projections could potentially be reduced by incorporating information on the prevalence of the infectious agent in persons incubating vCJD. The prospect of vCJD prevalence studies has been raised by detection of abnormal prion protein, thought to be the infectious agent, in appendices and tonsils removed from vCJD patients. Although unlinked anonymous testing of stored operative tissues for abnormal prion protein is very appealing, the design and interpretation of such prevalence studies is complicated by the lack of information on how early in the incubation period of vCJD the abnormal prion protein becomes detectable. METHODS: We simulate a range of vCJD epidemics, consistent with the limited available information on the incidence of vCJD, to illustrate some of the potential problems encountered when interpreting the results from prevalence studies of detectable abnormal prion protein. We assume plausible incubation period distributions and dietary exposure patterns. RESULTS: We demonstrate, in the context of our simulated epidemics, that prevalence studies of detectable abnormal prion protein would require the testing of tens of thousands of operative specimens and, even then, that unlinked anonymous testing positives would be unexpected.

Animals↗

Host-parasite relationships of Fasciola hepatica in the white mouse. VII. effects of anti-worm incubate sera on transferred worms and successful vaccination with a crude incubate antigen.

Mouse antisera against the 16-day-old worm incubate and sera from 25-day infections in mice debilitated migrating flukes in recipient animals as measured by worm recovery and host mortality. Mouse antisomatic and 100-day infection sera produced no such effects. Host mortality was significantly lower after challenge in mice given one ip immunizing injection of the worm incubate; however, worm recovery was not significantly reduced. Injections at 2, 7, 12, and 24 hr with the worm incubate elaborated over a 24-hr period protected 75% of the mice from infection after challenge, and reduced the worm burden by 83.3%.

Animals↗

Studies on sperm chromatin structure alterations and cytogenetic damage of mouse sperm following in vitro incubation. Studies on in vitro-incubated mouse sperm.

Mouse epididymal sperm incubated in Tyrode's T6 fertilization media were analyzed over time for chromosome damage by two methods. First, cytogenetic analysis was done on paternal pronuclei metaphase chromosomes. After 6 hours incubation 11% of the cells demonstrated chromosome structural abnormalities. Secondly, sperm nuclei were measured by the sperm chromatin structure assay, which is a measure of the susceptibility of sperm DNA to the nuclei demonstrated an increased susceptibility to DNA denaturation, reaching near 100% by 48 hours. Changes in chromatin structure at the molecular level may lead to chromosome breaks seen in pronuclear chromosomes.

Animals↗

A comparison of the effects of prior cold incubation on cerebral cortex function in a hibernator (Cricetus auratus) and a non-hibernator (Cavia porcellus)--II. High energy phosphate levels in cerebral cortex slices after in vitro cold incubation.

1. ATP and CP levels were measured in brain slices from golden hamster and guinea pig after varying periods of cold storage and subsequent incubation at 37 degrees C in the presence and absence of K+ salts. 2. ATP and CP levels were maintained at higher levels in hamster tissue. 3. The results are discussed in relation to the ability of a hibernator to transform and transport chemical energy at low temperatures.

Adenosine Triphosphate↗

Enhancement of varicella-zoster virus detection in A-549 shell vials by use of freeze-thawed specimens, extended incubation, and "a centrifuged, not incubated" direct detection method.

A total of 95 clinical samples were cultured for periods of 2 and 7 days in centrifuged A-549 shell vials before and after freezing and thawing of specimens. In addition, centrifuged A-549 shell vials were tested directly for varicella-zoster virus without incubation using a direct fluorescent antibody (DFA) technique. Twenty-seven specimens were positive by at least one method. The sensitivity for DFA on unincubated A-549 shell vials was 85.2%; for unfrozen 2-day cultures, 88.9%; for unfrozen 7-day cultures, 92.6%; for freeze-thaw 2-day cultures, 92.6%; and for freeze-thaw 7-day cultures, 96.3%. Freeze-thawed specimens cultured for 7 days yielded the highest number of positive results with conspicuous cell-to-cell spread as a sign of viral replication.

Cell Line↗

Unsaturated 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholines (unsaturated platelet-activating factor): aggregation of human platelets after incubation with indomethacin, creatinephosphate/creatinephosphokinase, xylocain and hirudine, and serotonin release after incubation with indomethacin.

Unsaturated platelet-activating factor (PAF) aggregates thrombocytes of healthy female volunteers and releases within 1 min up to 30.95% of the platelet serotonin. Indomethacin does not inhibit the aggregation but reduces the release of serotonin induced by unsaturated PAF in citrated platelet-rich plasma (PRP). Creatinephosphate combined with creatinephosphokinase (CP/CPK) inhibits the second phase, whereas xylocain inhibits the first and second phase of aggregation induced by unsaturated PAF. Hirudine shows no influence on the aggregation induced by unsaturated PAF.

Blood Platelets↗