Removal of ligand-bound liposomes from cell surfaces by microbubbles exposed to ultrasound

Gas-filled microbubbles attached to cell surfaces can interact with focused ultrasound to create microstreaming of nearby fluid. We directly observed the ultrasound/microbubble interaction and documented that under certain conditions fluorescent particles that were attached to the surface of live cells could be removed. Fluorescently labeled liposomes that were larger than 500 nm in diameter were attached to the surface of endothelial cells using cRGD targeting to αvβ3 integrin. Microbubbles were attached to the surface of the cells through electrostatic interactions. Images taken before and after the ultrasound exposure were compared to document the effects on the liposomes. When exposed to ultrasound with peak negative pressure of 0.8 MPa, single microbubbles and groups of isolated microbubbles were observed to remove targeted liposomes from the cell surface. Liposomes were removed from a region on the cell surface that averaged 33.1 μm in diameter. The maximum distance between a single microbubble and a detached liposome was 34.5 μm. Single microbubbles were shown to be able to remove liposomes from over half the surface of a cell. The distance over which liposomes were removed was significantly dependent on the resting diameter of the microbubble. Clusters of adjoining microbubbles were not seen to remove liposomes. These observations demonstrate that the fluid shear forces generated by the ultrasound/microbubble interaction can remove liposomes from the surfaces of cells over distances that are greater than the diameter of the microbubble.     

Onset of puberty in CD-1 mouse pups exposed prenatally through weaning to endophyte-infected tall fescue seed

This study was undertaken to investigate the effects of feeding endophyte - infected (Acremonium coenophialum ) tall fescue seed to CD-1 mouse dams (P(1)) during gestation and lactation, and on the subsequent growth and sexual maturity (onset of puberty) of their male and female offspring (F(1)). Forty-eight 21 d old pups (24 male and 24 female F(1) mice) were weaned from dams fed one of two diets containing 50% rodent chow (w/w) and 50% KY-31 tall fescue (Festuca arundinacea ) seed. The seed in Diet 1 was noninfected, while the seed in Diet 2 was 80% endophyte-infected. At weaning (21 d), the F(1) pups were fed rodent chow, ad libitum throughout the remaining experimental period. At 24 d, they were paired with sexually mature non-treated virgin CD-1 mice (fed 100% rodent chow) for one parturition cycle. Male F(1) mice were sacrificed at 84 d to determine testicular development. The age at the birth of the first litter for Diet 2 F(1) male (76.8 +/- 2.2 d) and female (58.4 +/- 2.1 d) was significantly greater (P<0.05) than the age at parturition for Diet 1 male and female F(1) test mice (64.1 +/- 1.8 and 51.9 +/- 1.2 d, respectively). At parturition, the female F(1) mice showed no significant differences (P>0.05) in either mean parturition weight or number of F(2) pups born per litter. However, total F(2) litter wight (11.38 +/- 1.14 g) and mean weight per F(2) pup (1.40 +/- 0.04 g) for Diet 2 female F(1) mice litters were lower (P<0.05) when compared with Diet 1 females (14.53 +/- 0.57 g and 1.66 +/- 0.02 g, respectively). No significant differences were observed between the two male F(1) treatment groups, for total F(2) litter weight or the number of pups born per F(2) litter. Although Diet 2 F(1) males weighed significantly less (P<0.05) at weaning and at pairing, final body weights at sacrifice (84 d) were not different (P>0.05) from the Diet 1 males.     

哺乳期棕色田鼠对非亲缘幼仔的行为反应

通过观察产后2 日(L2)、6 日(L6)、10 日(L10)、14 日(L14)和20 日(L20)的棕色田鼠母鼠对2 日龄、6 日龄、10 日龄和14 日龄非亲缘幼仔的攻击行为和衔回行为,探讨了哺乳期棕色田鼠的母性行为与非亲缘幼仔的发育(日龄或个体大小)及母鼠产后时间的关系。结果表明,在哺乳前期(L2,L6),非亲缘幼仔的日龄大小显著影响母鼠的攻击行为(P < 0.05);在哺乳中期(L10,L14)及后期(L20),非亲缘幼仔的日龄对母鼠的攻击行为没有显著影响(P > 0. 05)。在整个哺乳期,母鼠对14 日龄非亲缘幼仔的攻击频次最高(P < 0.05)。产后不同时间的母鼠对同一日龄非亲缘幼仔的攻击频次没有显著差异(P > 0.05)。这些结果表明,哺乳期内的棕色田鼠对非亲缘幼仔的攻击与幼仔的发育有关,与母鼠的产后时间无关。     

Production of mouse pups from germline transmission-failed knockout chimeras

Occasionally, chimeras do not transmit the gene of interest to pups in gene disruption experiments. However, the risk of failure could be reduced if we could identify embryonic stem (ES)-derived germ cells in the testis. Here, we report the production of pups from three lines of infertile chimeric male mice and the establishment of knockout lines by combining green fluorescent protein-tagged ES cells with intracytoplasmic sperm injection.     

Suckling deficits in rat pups exposed to alcohol in utero

With the aid of a pair-feeding procedure, two groups of pregnant Long-Evans rats were fed a liquid diet containing 35% or 0% ethanol-derived calories during days 6-20 of gestation. A third group was allowed free access to standard lab chow and water throughout pregnancy. At 6-7 or 9-10 days of age, suckling performance by male and female offspring representing the three prenatal treatment groups was examined. The test stimulus was a 6-10-day postparturient, anesthetized dam in which milk letdown was prevented. Compared to both pair-fed and lab chow controls, alcohol-exposed animals exerted a lower maximum suckling pressure, spent less time suckling during the test session, and displayed an altered suckling pattern. These data are consistent with existing clinical and experimental evidence documenting sucking deficits following prenatal alcohol exposure and are discussed in terms of prenatal alcohol-induced CNS impairment.     

Location and recognition of pups by adult female harp seals

Harp seal (Pagophilus groenlandicus) cows use visual, auditory and olfactory clues to locate and/or identify their pups. Identification is made at a close range and the pups are approached from a distance in a random manner. The topography of the ice supporting the pups changes frequently. These changes probably preclude or reduce the use of spatial memory by the adults. Because the pups cannot be identified at a distance, and because of the absence of stable landmarks, the adult must probably remain relatively close to her pup at all times. The large number of pups in the area suggests that random searching will not be efficient. The ability of various mammalian species to utilize spatial memory may well influence their modes of locating and identifying their young.     

Fast-twitch skeletal muscles of dystrophic mouse pups are resistant to injury from acute mechanical stress

Loss of the dystrophin-glycoproteincomplex from muscle sarcolemma in Duchenne's muscular dystrophy (DMD)renders the membrane susceptible to mechanical injury, leaky toCa²⁺, and disrupts signaling, but the precise mechanism(s)leading to the onset of DMD remain unclear. To assess the role ofmechanical injury in the onset of DMD, extensor digitorum longus (EDL)muscles from C57 (control), mdx, andmdx-utrophin-deficient [mdx:utrn(/); dystrophic] pups aged 9-12 days were subjected to an acutestretch-injury or no-stretch protocol in vitro. Before the stretches,isometric stress was attenuated for mdx:utrn(/) comparedwith control muscles at all stimulation frequencies (P < 0.05). During the stretches, EDL muscles for each genotypedemonstrated similar mean stiffness values. After the stretches,isometric stress during a tetanus was decreased significantly for bothmdx and mdx:utrn(/) muscles compared withcontrol muscles (P < 0.05). Membrane injury assessedby uptake of procion orange dye was greater for dystrophic comparedwith control EDL (P < 0.05), but, within eachgenotype, the percentage of total cells taking up dye was not different for the no-stretch vs. stretch condition. These data suggest that thesarcolemma of maturing dystrophic EDL muscles are resistant to acutemechanical injury.

     

The ultrasonic responses of albino mouse pups to tactile stimuli

Tactile stimuli, like environmental temperature changes, can evoke ultrasonic responses from albino mouse pups. But the changes with age in the intensity of ultrasounds so produced follow a different pattern from those due to temperature changes. The responses begin with very high intensity pulses in the very young pups and then gradually decline with the age of the pups. The present report arises from a systematic study of this phenomenon, the results of which are discussed in relation to those of previous ones.     

Induction of maternal nest building in virgin female mice by the presentation of young

Ovariectomized and intact adult virgin female mice presented with two 1-day-old mouse pups built larger nests and more nests rated “maternal” than did intact animals not proffered young. Virgin females presented with live 1-day olds behind a wire partition in their homecages also constructed larger nests and more nests rated “maternal” as compared to animals presented with freshly killed 1-day olds, 19-day olds, or with no pups behind the partition.     

Effects of ultrasound on the mouse exposed at different stages of gestation: Acute studies

Summary At five stages of gestation, mice were exposed dorsally to ultrasonic irradiation at 1 W/cm² (spatial and temporal average), 2 MHz, and 34 and 37° C (temperature at beginning of exposure). The results indicated that ultrasonic exposure up to at least 100 s is not hazardous to a pregnant mouse or its offspring in utero under the conditions of our experiment. Thresholds for both undesirable and lethal effects on the dam and its embryos or fetuses appeared between 100 and 200 s. Temperature rises in the uterus to more than 40° C and similarity of results obtained with continuous-wave and burst modes of irradiation suggest a thermal mechanism as one cause of these effects. Preliminary conclusions are that thresholds for both tissue damage to the dam and deleterious effects on pregnancy were similar; however, they occurred at spatial average intensity and exposure levels far in excess of those used in clinical diagnostic medicine.     

Survival of cultured mammalian cells exposed to ultrasound

Summary The colony-forming ability of cultured mammalian cells exposed to monochromatic ultrasonic vibrations of different frequencies (0.1, 0.5, 1.0, 2.0, 3.3 MHz) was studied. The combined effect of x rays and 1.0-MHz ultrasonic waves on the survival of M3-1 cells was also investigated. Split-dose experiments showed that cells exposed to ultrasound are sensitized to a subsequent exposure. Almost twice as many cells survive a given ultrasonic dose when exposed in the M phase as when exposed in the S phase. A small amount of synergism between ultrasound and x rays has been observed. The extent of synergism depends on the experimental factors, and may be due to an interaction between nuclear damage caused by x rays and the damage to the cell membrane caused by ultrasound.Research supported by the U.S. Atomic Energy Commission and the National Aeronautics and Space Administration     

Chromosomic aberrations in female workers exposed to pesticides

The purpose of this work was to determine if the occupational exposure to those pesticides used at banana plantations' packaging plants produces genetic damage to somatic cells of female workers. Chromosomal aberrations were scored in lymphocytes of 20 women, 10 female exposed workers and 10 female controls. Workers were recruited from independent farms from two locations in Costa Rica, during January through June in 1996 and 1997. These females had a minimum of three months of work, had never received chemotherapy or radiotherapy and did some of these labors: sealing, spraying or weighting of bananas. Control unexposed females lived in the same area, were of similar age and neither them nor their husbands/mates had ever worked in pesticide related labors. For each female, 100 mitotic figures were scored. The kind of aberrations detected were acentric fragments, dicentric chromosomes, rings, gaps and breaks. Among workers, 16% of cells (n=1000) had one or more abnormalities, whereas control unexposed females had 6% of cells (n=1000) with comparable anomalies (p < 0.05). In conclusion, the pesticide exposure is a risk factor for chromosome aberrations in female somatic cells.     

Pregnancy in female house mice exposed to urinary chemosignals from other females.

Seven experiments were performed to investigate pregnancy termination, urinary chemosignals, and litter sex ratio variation in female house mice. Experiments tested the effects of urine from adult and prepubertal females, housed individually or in groups, on successful insemination and litter production by females treated at different times and for different periods during the 3 weeks before mating and during gestation. Treatment of females with urine from adult females housed eight per cage or with urine pooled from eight adult females housed individually for 2 or 3 weeks before mating resulted in fewer successful pregnancies and significantly more female-biased litters. Treatment with urine from adult or prepubertal females housed eight per cage or with urine pooled from eight mice housed individually for the first 6 days of gestation or throughout pregnancy resulted in a significant increase in the rate of pregnancy termination. These treatments resulted in lower body weights at birth and slower growth rates in all males and in some females. Puberty was delayed in female progeny from urine-treated dams in five of seven experiments, and these young females attained first oestrus at greater mean body weights than mice in other treatments. These findings indicate that, in mice, at high population density, communication via a urinary chemosignal can alter reproduction in recipient females. Availability of, and competition for, resources such as food would be greater at higher densities, possibly lowering the probability of reproductive success. Pregnancy termination and delays in reproduction and attainment of sexual maturity might lead to greater successful reproduction at a later time.