Effects of low versus physiologic plasma progesterone concentrations on ovarian follicular development and fertility in beef cattle.

The objective of this study was to determine the effects of low versus physiologic plasma progesterone concentrations during the ovulatory wave on fertility in cattle. Suckled beef cows (Bos taurus; n=129) and pubertal heifers (Bos taurus; n=150) at random stages of the estrous cycle were given a luteolytic dose of prostaglandin F(2 alpha) (500 microg cloprostenol; PGF) twice, 11 d apart. Ten days after the second PGF treatment, cattle were given estradiol benzoate im (1.5 and 1.0mg for cows and heifers, respectively) and a progesterone-releasing intravaginal device (Cue-Mate) with a single pod containing 0.78 g progesterone (Day 0). Cattle in the low-progesterone group (n = 148) received a luteolytic dose of PGF on Day 0, whereas those in the high-progesterone (i.e., physiologic plasma concentrations) group (n=131) were allowed to retain their corpora lutea. On Day 8, the Cue-Mate was removed, and PGF was given to both groups. Fifty-four hours to 56 h later, cattle received 12.5mg of porcine LH (pLH) im and were concurrently artificially inseminated. The dominant follicle in the low-progesterone group was larger (P<0.001) than that in the high-progesterone group on the day of insemination (14.9+/-0.3mm vs. 12.7+/-0.3mm, mean+/-SEM). At 7 d after ovulation, the low-progesterone group had a larger corpus luteum (24.5+/-0.54 mm vs. 21.9+/-0.64 mm, P<0.01) and higher plasma progesterone concentration (4.0+/-0.3 vs. 3.1+/-0.2, P<0.01) than that of the high-progesterone group. However, pregnancy rates did not differ (79 of 148, 53.4%, and 70 of 131, 53.4%) for low- and high-progesterone groups, respectively). In summary, low circulating progesterone concentrations during the growing phase of the ovulatory follicle resulted in a larger dominant follicle and a larger CL that produced more progesterone, with no significant effect on pregnancy rate.

Effects of progesterone presynchronization and eCG on pregnancy rates to GnRH-based, timed-AI in beef cattle.

Three experiments were conducted to determine the effects of low-dose progesterone presynchronization and eCG on pregnancy rates to GnRH-based, timed-AI (TAI) in beef cattle (GnRH on Day 0, PGF(2alpha) on Day 7, with GnRH and TAI on Day 9, 54-56 h after PGF(2alpha)). Experiments 1 and 2 were 2 x 2 factorials with presynchronization (with or without a once-used CIDR; Days -15 to 0 in Experiment 1 and Days -7 to 0, with PGF(2alpha) at insertion, in Experiment 2), and with or without 400 IU eCG on Day 7 in suckled cows. In Experiment 3, suckled cows and nulliparous heifers were either presynchronized with a twice-used CIDR (Days -5 to 0) and PGF(2alpha) at insertion, or no treatment prior to insertion of a new CIDR (Days 0-7). Presynchronization increased (P<0.05) ovulation rate to GnRH on Day 0 (75.0% vs 48.7%, 76.7% vs 55.0%, and 60.0% vs 36.1% for Experiments 1, 2, and 3, respectively), increased the diameter of the preovulatory follicle in Experiments 1 and 2, and increased the response to PGF(2alpha) (regardless of parity) in Experiment 1 (P<0.01), and in primiparous cows in Experiment 2 (P<0.01). Effects of presynchronization on pregnancy rates (53.4% vs 54.1%, 57.7% vs 45.3%, and 54.3% vs 44.4% for Experiments 1, 2, and 3, respectively) were influenced by parity and eCG (P<0.05). Treatment with eCG had no effect (P>0.05) on the diameter of the preovulatory follicle (Experiment 1), or the response to PGF(2alpha) (Experiments 1 and 2), but tended (P=0.08) to improve pregnancy rates, especially in primiparous cows that were not presynchronized (P<0.01). However, the effects of eCG and presynchronization were not additive.

Lower pregnancy losses in lactating dairy cows fed a diet enriched in alpha-linolenic acid.

The objectives were to determine if a diet enriched in alpha-linolenic acid (ALA) would influence ovarian function, early embryo survival, conception rates, and pregnancy losses in lactating dairy cows. Beginning 28 d before breeding, Holstein cows (55 +/- 22 d postpartum; mean +/- SD) were assigned to diets supplemented with either rolled flaxseed (FLAX; 56.7% ALA, n = 62) or rolled sunflower seed (SUNF; 0.1% ALA, n = 59) to provide approximately 750 g of oil/d. Diets continued for 32 d after timed artificial insemination (TAI, d 0) following a Presynch/Ovsynch protocol. Barley silage- and barley grain-based TMR were formulated to meet or exceed National Research Council requirements. Metabolizable protein and net energy for lactation concentrations were similar in the 2 diets. Based upon a mean dry matter intake of 22 kg/d, cows fed FLAX or SUNF consumed > 410 g or < 1 g of ALA, respectively. Pregnancy was confirmed by ultrasound 32 d after TAI. Nonpregnant cows were placed on a second Ovsynch regimen and reinseminated 42 d after first TAI, and received oilseeds for 32 d after second TAI. Relative to prediet levels, FLAX increased the ALA content of milk by 187%. Ovarian ultrasonography was performed in 8 cows per diet; the mean diameter of ovulatory follicles was larger in cows fed FLAX compared with SUNF (16.9 +/- 0.9 vs. 14.1 +/- 0.9 mm), but follicle number, corpus luteum size, and plasma progesterone concentrations remained unaffected. Presumptive conception (progesterone < 1 ng/mL on d 0 and > 1 ng/mL on d 21) rates to first TAI were greater in FLAX than in SUNF (72.6 vs. 47.5%). Pregnancy losses were lower in cows fed FLAX (9.8%) compared with those fed SUNF (27.3%). Including flaxseed in the ration of dairy cows increased the size of the ovulatory follicle and reduced pregnancy losses.

Resynchronization of previously timed-inseminated beef heifers with progestins.

The objective was to determine the efficacy of a previously used CIDR or melengestrol acetate (MGA; 0.5mg/head/day) for resynchronization of estrus in beef heifers not pregnant to timed-AI (TAI). In three experiments and a field trial, heifers were reinseminated 6-12 h after first detection of estrus. Pregnancy diagnosis was done from approximately 25-43 days after either TAI or reinsemination. In Experiment 1, 79 heifers received a once-used CIDR from 13 to 20 days after TAI and 80 heifers were untreated controls. For these two groups, there were 34 and 35 heifers, respectively, not pregnant to TAI; median +/- S.E. intervals from TAI to onset of estrus were 22 +/- 0.2 days versus 20 +/- 0.6 days (P < 0.001); estrus rates were 70.6% versus 85.7% (P = 0.1); conception rates were 62.5% versus 76.7% (P < 0.3); and pregnancy rates were 44.1% versus 65.7% (P = 0.07), for CIDR and untreated (control) groups, respectively. In Experiment 2, heifers (n = 651) were TAI (Day 0) and 13 days later randomly assigned to one of seven groups (n = 93 per group) to receive a once-used CIDR (three groups; Days 13-20), MGA (three groups; Days 13-19), or no treatment (control group). Groups given a CIDR or MGA also received: no further treatment (CIDR or MGA alone); 1.5mg estradiol-17beta (E-17beta) and 50 mg progesterone (P4) in 2 mL canola oil on Day 13; or E-17beta and P4 on Day 13 and 0.5 mg E-17beta on Day 21 (24 h after CIDR removal or 48 h after the last feeding of MGA). Pregnancy rate to TAI was lowest (P < 0.05) for the group given a CIDR plus E-17beta and P4 on Day 13 and E-17beta on Day 21. Variability in return to estrus was greater (P < 0.001) in the control and MGA groups than in CIDR groups. Conception and pregnancy rates in heifers given a CIDR (65.1 and 61.4%) were higher (P<0.01) than those fed MGA (49.6 and 40.4%), but not different from controls (62.2 and 54.9%, respectively). In Experiment 3, 616 heifers received a once- or twice-used CIDR for 7 days, beginning 13+/-1 days after TAI, with or without a concurrent injection of 150 mg of P4 (2 x 2 factorial design). Pregnancy rate to TAI was 47.2%. In heifers that returned to estrus, there was no significant difference between a once- or twice-used CIDR for rates of estrus (68.8%, P < 0.3), conception (65.9%, P < 0.6) and pregnancy (45.3%, P < 0.8). Injecting progesterone at CIDR insertion increased the median interval from CIDR removal to onset of estrus (P < 0.05) and reduced rates of estrus (63.8% versus 73.8%, P<0.05), conception (60.5% versus 70.6%, P = 0.1) and pregnancy (38.6% versus 52.2%, P < 0.02). In a field trial, 983 heifers received a once-used CIDR for 7 days, beginning 13 +/- 1 days after TAI. Pregnancy rate to TAI was 55.2%. The median (and mode) of the interval from CIDR removal to estrus was 2.5 days. Estrus, conception and pregnancy rates were 78.2, 70.3 and 55.0% (overall pregnancy rate to TAI and rebreeding, 78.7%). In summary, a once- or twice-used CIDR for 7 days, starting 13 +/- 1 days after TAI resulted in the majority of nonpregnant heifers detected in estrus over a 4-day interval, with acceptable conception rates; however, injecting progesterone at CIDR insertion significantly reduced both estrus and pregnancy rates, and estradiol treatment after CIDR removal was associated with a decreased pregnancy rate to TAI. Fertility was higher in heifers resynchronized with a once-used CIDR than with MGA.

Effect of estradiol valerate on ovarian follicle dynamics and superovulatory response in progestin-treated cattle.

Three experiments evaluated the effects of estradiol valerate (EV) on ovarian follicular and CL dynamics, intervals to estrus and ovulation, and superovulatory response in cattle. Experiment 1 compared the efficacy of two norgestomet ear implants (Crestar and Syncro-Mate B; SMB) for 9 d (with PGF at implant removal), combined with either 5 mg estradiol-17beta and 100 mg progesterone (EP) or 5 mg EV and 3mg norgestomet (EN) im at the time of implant insertion on CL diameter and follicular wave dynamics. Ovaries were monitored by ultrasonography. There was no effect of norgestomet implant. Diameter of the CL decreased following EN treatment (P < 0.01). Mean (+/- S.D.) day of follicular wave emergence (FWE) was earlier (P < 0.0001) and less variable (P < 0.0001) in EP- (3.6 +/- 0.5 d) than in EN- (5.7 +/- 1.5 d) treated heifers. Intervals from implant removal to estrus (P < 0.001) and ovulation (P < 0.01) were shorter in EN- (45.7 +/- 11.7 and 74.3 +/- 12.6 h, respectively) than in EP- (56.4 +/- 14.1 and 83.3 +/- 17.0 h, respectively) treated heifers. Experiment 2 compared the efficacy of EP versus EN in synchronizing FWE for superovulation in SMB-implanted cows. At random stages of the estrous cycle, Holstein cows (n = 78) received two SMB implants (Day 0) and were randomly assigned to receive EN on Day 0 or EP on Day 1. Folltropin-V treatments were initiated on the evening of Day 5, with PGF in the morning and evening of Day 8, when SMB were removed. Cows were inseminated after the onset of estrus and embryos were recovered 7 d later. Non-lactating cows had more CL (16.7 +/- 11.3 versus 8.3 +/- 4.9) and total ova/embryos (14.7 +/- 9.5 versus 7.9 +/- 4.6) than lactating cows (P < 0.05). EP-treated cows tended (P = 0.09) to yield more transferable embryos (5.6 +/- 5.2) than EN-treated cows (4.0 +/- 3.7). Experiment 3 compared the effect of dose of EV on ovarian follicle and CL growth profiles and synchrony of estrus and ovulation in CIDR-treated beef cows (n = 43). At random stages of the estrous cycle (Day 0), cows received a CIDR and no further treatment (Control), or an injection of 1, 2, or 5 mg im of EV. On Day 7, CIDR were removed and cows received PGF. Follicular wave emergence occurred within 7 d in 7/10 Control cows and 31/32 EV-treated cows (P < 0.05). In responding cows, interval from treatment to FWE was longer (P < 0.05) in those treated with 5 mg EV (4.8 +/- 1.2 d) than in those treated with 1 mg (3.2 +/- 0.9 d) or 2 mg (3.4 +/- 0.8 d) EV, while Control cows were intermediate (3.8 +/- 2.0 d). Diameter of the dominant follicle was smaller (P < 0.05) at CIDR removal and tended (P = 0.08) to be smaller just prior to ovulation in the 5 mg EV group (8.5 +/- 2.2 and 13.2 +/- 0.6 mm, respectively) than in the Control (11.8 +/- 4.6 and 15.5 +/- 2.9 mm, respectively) or 1mg EV (11.7 +/- 2.5 and 15.1 +/- 2.2 mm, respectively) groups, with the 2mg EV group (10.7 +/- 1.5 and 14.3 +/- 1.7 mm, respectively) intermediate. Diameter of the dominant follicle at CIDR removal was less variable (P < 0.01) in the 2 and 5mg EV groups than in the Control group, and intermediate in the 1mg EV group. In summary, treatment with 5mg EV resulted in a longer and more variable interval to follicular wave emergence than treatment with 5mg estradiol-17beta, which affected preovulatory dominant follicle size following progestin removal, and may have also affected superstimulatory response in Holstein cows. Additionally, 5 mg EV appeared to induce luteolysis in heifers, reducing the interval to ovulation following norgestomet removal. Conversely, intervals to, and synchrony of, follicular wave emergence, estrus and ovulation following treatment with 1 or 2 mg EV suggested that reduced doses of EV may be more useful for the synchronization of follicular wave emergence in progestogen-treated cattle.

Improving the quality of electron backscatter diffraction (EBSD) patterns from nanoparticles.

In this study, we investigated the relative contributions of atomic number (Z) and density (rho) to the degradation of the electron backscatter diffraction (EBSD) pattern quality for nanoparticles < 500 nm in diameter. This was accomplished by minimizing the diffuse scattering from the conventional thick mounting substrate through the design of a sample holder that can accommodate particles mounted on thin-film TEM substrates. With this design, the contributions of incoherently scattered electrons that result in the diffuse background are minimized. Qualitative and quantitative comparisons were made of the EBSD pattern quality obtained from Al(2)O(3) particles approximately 200 nm in diameter mounted on both thick- and thin-film C substrates. For the quantitative comparison we developed a 'quality' factor for EBSD patterns that is based on the ratio of two Hough transforms derived from a given EBSD pattern image. The calculated quality factor is directly proportional to the signal-to-noise ratio for the EBSD pattern. In addition to the comparison of the thick and thin mounting substrates, we also estimated the effects of Z and rho by comparing the EBSD pattern quality from the Al(2)O(3) particles mounted on thin-film substrates with the quality of patterns obtained from Fe-Co nanoparticles approximately 120 nm in diameter. The results indicate that the increased background generated in EBSD patterns by the electrons escaping through the bottom of the small particles is the dominant reason for the poor EBSD pattern quality from nanoparticles < 500 nm in size. This was supported by the fact that we were able to obtain usable EBSD patterns from Al(2)O(3) particles as small as 130 nm using the thin-film mounting method.

The Analysis of Particles at Low Accelerating Voltages (≤ 10 kV) With Energy Dispersive X-Ray Spectroscopy (EDS).

In recent years, there have been a series of advancements in electron beam instruments and x-ray detectors which may make it possible to improve significantly the quality of results from the quantitative electron-probe analysis of individual particles. These advances include: (1) field-emission gun electron beam instruments such as scanning electron microscopes (FEG-SEMs) that have high brightness electron guns with excellent performance at low beam energies, E 0 ≤ 10 keV and (2) high-resolution energy-dispersive x-ray spectrometers, like the microcalorimeter detector, that provide high-resolution (< 10 eV) parallel x-ray collection. These devices make it possible to separate low energy (< 4 keV) x-ray lines including the K lines of carbon, nitrogen and oxygen and the L and M lines for elements with atomic numbers in the range of 25 to 83. In light of these advances, this paper investigates the possibility of using accelerating voltages ≤ 10 kV, as a method to improve the accuracy of elemental analysis for micrometer-sized particles.

Phase identification of individual crystalline particles by electron backscatter diffraction.

Recently, an electron backscatter diffraction (EBSD) system was developed that uses a 1024 x 1024 CCD camera coupled to a thin phosphor. This camera has been shown to produce excellent EBSD patterns. In this system, crystallographic information is determined from the EBSD pattern and coupled with the elemental information from energy or wavelength dispersive X-ray spectrometry. Identification of the crystalline phase of a sample is then made through a link to a commercial diffraction database. To date, this system has been applied almost exclusively to conventional, bulk samples that have been polished to a flat surface. In this investigation, we report on the application of the EBSD system to the phase identification analysis of individual micrometre and submicrometre particles rather than flat surfaces.

Sentence repetition and processing resources in Alzheimer's disease.

Sentence processing in Alzheimer's disease (AD) has been found to be influenced by several grammatical and extragrammatical factors, including phrase structure and verb-argument relations, number of propositions/verbs, and processing resource capacity. This study examines the effects of these variables on sentence production in AD. Normal control and AD subjects were asked to repeat six types of sentences varying along the above dimensions of complexity. Subjects' processing resource capacity was measured using several verbal working memory tests. AD subjects' sentence-repetition performance was impaired compared to the normal control group. Significant effects were observed for branching direction of phrase structure, canonicity of verb-argument relations, and serial position of errors. Sentence-repetition performance significantly correlated with working memory scores. The findings are interpreted within a resource capacity theory of sentence processing.

Measurement of vaginal temperature by radiotelemetry for the prediction of estrus in beef cows.

Peaks in vaginal temperature were assessed as predictors of estrus in 22 suckled beef cows during 2 breeding seasons. A flexible plastic anchor was used to retain a temperature radiotransmitter within the cow vagina and vaginal temperature was monitored at 4-min intervals. Blood was collected twice weekly for analysis of serum progesterone to confirm the occurrence of estrus and ovarian status was checked weekly using transrectal ultrasonography. Visual observations of estrous behavior were made for 20 min/h between 0400 and 0800 h and casually from 0800 to 1600 h (4 to 6 times). Values for vaginal temperature were available for 47 estrous periods. The prediction of estrus based on vaginal temperature was excellent when an estral peak in vaginal temperature was defined as an increase of at least 0.4 degree C for 3 or more consecutive hours using the corresponding hourly means of a 2 or 3-d baseline. Combining the results of 2 separate years and using a 3-d baseline, a peak in vaginal temperature was found for 42 of 47 confirmed estrus periods (detection sensitivity of 89.4% and prediction power positive of 85.7%). Corresponding detection sensitivity and prediction power positive for visual observations of standing estrus were 53.2 and 96.2%, respectively. The mean maximal increase in vaginal temperature at estrus was 0.9 +/- 0.3 degree C and the mean duration of the estrual peak in vaginal temperature was 6.5 +/- 2.7 h. In addition, vaginal temperature was found to be significantly depressed for 3 d prior to estrus and significantly elevated at mid-cycle.

Sentence comprehension in Alzheimer's disease: effects of grammatical complexity, speech rate, and repetition.

Caregivers of patients diagnosed with Alzheimer's disease (AD) are often advised to modify their speech to facilitate the patients' sentence comprehension. Three common recommendations are to (a) speak in simple sentences, (b) speak slowly, and (c) repeat one's utterance, using the same words. These three speech modifications were experimentally manipulated in order to investigate their individual and combined effects on sentence comprehension in AD. Fifteen patients with mild to moderate AD and 20 healthy older persons were tested on a sentence comprehension task with sentences varying in terms of (a) degree of grammatical complexity, (b) rate of presentation (normal vs. slow), and (c) form of repetition (verbatim vs. paraphrase). The results indicated a significant decline in sentence comprehension for the AD group. Sentence comprehension improved, however, after the sentence was repeated in either verbatim or parapharsed form. However, the patients' comprehension did not improve for sentences presented at the slow speech rate. This pattern of results is explained via-à-vis the patients' working memory loss. The findings challenge the appropriateness of several clinical recommendations.

Grammatical abilities in Parkinson's disease: evidence from written sentences.

This study examined the grammatical content of written sentences elicited from 96 Parkinson's patients, 30 Parkinson's with dementia patients and 167 control subjects. Parkinson's patients without dementia or with mild dementia presented no impairments in sentence length, syntactic complexity or amount of information content. Moderately demented Parkinson's patients showed reduced sentence length and information content but normal syntactic complexity. This pattern of results provides evidence that lexical-semantic content is more susceptible to decline than syntactic structure with the progression of dementia in Parkinson's disease.

Research on neuropsychiatric genetics: interests of the National Institute of Neurological Disorders and Stroke.

The National Institute of Neurological Disorders and Stroke (NINDS) supports research concerning the determinants of normal and pathological development of the nervous system, from the genetic to the environmental. NINDS also funds basic and clinical research concerning the etiology, diagnosis, treatment, consequences, and prevention of the spectrum of neurodevelopmental disorders including neurobehavioral and neurodegenerative disorders (National Advisory Council Neurological Disorders and Stroke, 1989, 1992; Division of Convulsive, Developmental and Neuromuscular Disorders, NINDS, 1993).

Biochemical and cellular properties of three immortalized Schwann cell lines expressing different levels of the myelin-associated glycoprotein.

Biochemical and cellular properties of three immortalized Schwann cell lines expressing different levels of the myelin-associated glycoprotein (MAG) were compared. The S16 line generated by repetitive passaging was described previously and expresses a level of MAG comparable to that in adult sciatic nerve. The S42 line was generated independently by the same procedure, divides more slowly than the S16 line, and expresses an even higher level of MAG. The S16Y line arose spontaneously from a passage of the S16 cells, divides much more rapidly, and does not express MAG. The levels of MAG expression in the three lines are inversely related to their rates of proliferation, and MAG mRNA levels parallel the amounts of MAG. The S16 and S42 lines consist mainly of flat cells at low density and develop many processes at high density, whereas most of the S16Y cells are spindle-shaped, resembling primary Schwann cells in appearance. Surface immunostaining with the O4 antibody was positive for the S16 and S42 cells and negative for the S16Y cells, but all three lines were negative for surface staining with the O1 antibody. The overall protein compositions of the three lines are very similar, but the S16 and S42 cells express larger amounts of several glycoproteins than the S16Y cells, including the adhesion proteins, neural cell adhesion molecule, L1, and laminin. S16 and S42 cells (but not S16Y cells) also express P0 glycoprotein, galactocerebroside, and sulfatide, but, unlike MAG, these other myelin-related components were present at much lower levels than in adult nerve. Myelin basic protein and proteolipid protein were not detected in any of the lines, although all three lines contained proteolipid protein mRNA. 2',3'-Cyclic nucleotide 3'-phosphodiesterase and glial fibrillary acidic protein were present in all three lines. Conditions have not yet been found in which any of the lines will myelinate dorsal root ganglion neurons in vitro, but the S16 and S42 cells differ from the S16Y cells by clustering around neurons after 1 week in coculture. In many respects, the S16 and S42 cells biochemically resemble Schwann cells at an early stage in their preparation to myelinate and should be useful for investigating the cell biology of MAG and other myelin-related components.

Sequences in the visna virus long terminal repeat that control transcriptional activity and respond to viral trans-activation: involvement of AP-1 sites in basal activity and trans-activation.

Visna virus is a pathogenic lentivirus of sheep whose gene expression is developmentally regulated in cells of the monocyte-macrophage lineage. Gene expression directed by the visna virus long terminal repeat (LTR) is increased in infected cells by a virus-encoded trans-acting protein. trans-Activation is mediated in part by increases in the steady-state level of mRNA. Deletion and linker-scanner mutants were constructed to locate sequences in the LTR that regulate transcription and are responsive to viral trans-activation. The activities of these mutants were tested by using them to drive transcription of the bacterial gene for chloramphenicol acetyltransferase in transient expression assays. Three regions located between-140 and the cap site were found to be important for basal transcriptional activity, and the importance of each region was found to be dependent on the cell type. Sequences responsive to viral trans-activation were found to be the same sequences required for basal transcriptional activity. The visna virus LTR contains six sequences that are homologous to the recognition site for cellular transcriptional factor AP-1 and a single sequence homologous to the recognition site for transcriptional factor AP-4. Both of these classes of binding sites appear to be important for regulating the basal level of transcription of visna virus. The AP-1-binding site most proximal to the TATA box was found to be one target for viral trans-activation. The visna virus promoter was found to be activated by serum; this serum response has also been mapped to the AP-1-related sequences in the LTR.

Regulation of the visna virus long terminal repeat in macrophages involves cellular factors that bind sequences containing AP-1 sites.

Visna virus gene expression is highly restricted in monocytes but is induced by monocyte-macrophage differentiation in vivo. Deletion and linker-scanning mutants, gel shift assays, and DNase I footprinting were used to identify sequences in the visna virus long terminal repeat involved in the developmental regulation of gene expression in the U937 monocytic cell line. We found that an AP-1 and an AP-4 binding site were critical for basal activity and that the AP-1 site was required for phorbol ester-inducible gene expression. These results suggest that cellular factors that interact with AP-1 sites are involved in the developmental regulation of visna virus gene expression in macrophages.

The visna virus long terminal repeat directs expression of a reporter gene in activated macrophages, lymphocytes, and the central nervous systems of transgenic mice.

Visna virus is a lentivirus which causes a slow progressive disease involving the immune system and the central nervous system. To determine the role of the viral long terminal repeat (LTR) in targeting the virus to specific host cells and tissues, transgenic mice were constructed which contained the visna virus LTR directing expression of the bacterial gene encoding chloramphenicol acetyltransferase (CAT). Analysis of the transgenic mouse tissues for CAT activity revealed that the viral LTR was responsible, in part, for the tropism of visna virus for macrophages and the central nervous system. Expression of the LTR required the macrophage to be in an activated state both in vivo and in vitro. Thioglycolate activation of peritoneal macrophages in vivo and 12-O-tetradecanoylphorbol 13-acetate treatment in vitro induced expression of the visna virus LTR. Lymphocytes from the spleens of the transgenic mice expressed CAT activity, suggesting that visna virus was able to replicate in lymphocytes, as did human immunodeficiency virus and simian immunodeficiency virus. These studies demonstrated that the lentivirus LTR was responsible, in part, for cell and tissue tropism in vivo.

Dysmyelination in transgenic mice containing JC virus early region.

JC virus (JCV) causes the chronic human demyelinating disease progressive multifocal leukoencephalopathy. Because of host range restrictions, experimental models of JCV-induced demyelination have not been available. The restricted tropism of JCV infectivity has recently been overcome by the production of transgenic mice that contain the early region of JCV in all cells. This portion of the DNA encodes JCV T-antigens. These mice display a dysmyelinating phenotype, the severity of which is related to the level of JCV early region expression in brain. With the use of immunocytochemistry and in situ hybridization, we characterized morphologically myelin-specific and JCV gene expression in a severely affected strain of these mice. Our results suggest that expression of JCV T-antigens occurs predominantly in oligodendrocytes and is the primary cause of dysmyelination. Affected oligodendrocytes do not myelinate axons properly. However, they express myelin-specific genes and display some of the morphological phenotypes of early stages of myelination. A decreased ratio between levels of transcriptional and translational products of genes encoding the major structural proteins of central nervous system myelin was apparent. These results suggest that JCV T-antigens arrest the maturation of oligodendrocytes and inhibit the production of myelin. These results also demonstrate that JCV transgenic mice are a good model for investigating mechanisms of JCV-induced demyelinating lesions in progressive multifocal leukoencephalopathy.

Early regions of JC virus and BK virus induce distinct and tissue-specific tumors in transgenic mice.

JC virus and BK virus are ubiquitous human viruses that share sequence and structural homology with simian virus 40. To characterize tissue-specific expression of these viruses and to establish model systems for the study of human viral-induced disease, transgenic mice containing early regions of each of the viruses were produced. The viral sequences induced tumors in a distinct and tissue-specific manner that was similar to their tissue tropism in humans. Ten JC virus-containing founder mice were produced, of which 5 survived to maturity. Four of them developed adrenal neuroblastomas, which metastasized to several other tissues. JC virus tumor-antigen RNA was detected at high levels in the tumor tissues and at low levels in the normal tissues of these mice. One of the three BK virus-containing mice was abnormally shaped and died at 2 weeks of age. The other two BK virus-containing mice developed primary hepatocellular carcinomas and renal tumors and died at 8-10 months of age. BK virus tumor-antigen RNA was expressed in tumor tissues of both mice. Since each of the viruses retained the general tissue tropism that it exhibits in humans, these data suggest that transgenic mice harboring human viruses will be useful as animal models for viral-induced diseases.