Matua bromegrass hay for mares in gestation and lactation.

Matua bromegrass hay (Bromus willdenowii Kunth) is a high quality forage, but its value for mares during gestation and lactation is not well known. Intake, rate of passage, performance, and reproduction by gestating and lactating Quarter Horse mares fed the hay was investigated. In this experiment, 12, 2- to 12-yr-old gravid mares (mean BW = 553 kg, SD = 36) were fed Matua hay (CP = 11.5%) or alfalfa hay (Medicago sativa L.) (CP = 15.4%) for variable days prepartum (mean 59.9 d; SD = 23.5) and for 70 d postpartum. Matua and alfalfa hay were fed as the roughage portion of the diet with a grain supplement. Mares, blocked by age, expected date of foaling, and BW, were assigned randomly within blocks to treatments (six mares per treatment). Forage type did not affect intake, gestation length, birth weight, number of foals, foal weight gain, day of first postpartum ovulation, cycles per conception, or pregnancy rate at 70 d. On d 1, milk from mares fed alfalfa hay contained less (P < 0.03) CP than milk from mares fed Matua hay. Milk CP decreased (P < 0.01) in all mares over time. In a separate experiment, voluntary intake and rate of passage of Matua (CP = 15.5%), alfalfa (CP = 24.9%), and Timothy (Phleum pratense L.) (CP = 4.1%) hays were determined in nine 2-yr-old pregnant mares (mean BW = 447 kg; SD = 21). Diets were 100% forage. Timothy hay did not meet CP requirements for mares. Voluntary intake of alfalfa hay was higher (P < 0.01) than Matua hay. Intake of Timothy hay was lower (P < 0.01) than the mean of alfalfa and Matua hay. Rate of passage offorage was measured by passage of Cr-mordanted fiber. Passage rate and retention time did not differ between Matua and alfalfa hay; however, the retention times of Matua and alfalfa hays were shorter (P < 0.01) than for Timothy hay. Our results indicate that Matua hay is a forage that can be used safely for mares during gestation and early lactation and for their young foals.

Voluntary intake, digestibility, and subsequent selection of Matua bromegrass, coastal bermudagrass, and alfalfa hays by yearling horses.

Matua bromegrass (Bromus willdenowii Kunth. cv. Grasslands Matua) was introduced in 1973, but little information exists concerning its potential as a hay for horses. Thus, voluntary intake and apparent digestibility of OM, CP, and fiber components of Matua by 18 Quarter Horse yearlings (mean initial BW 354 kg; SE 5.8) were compared with alfalfa (Medicago sativa L.) and coastal bermudagrass (Cynodon dactylon L.) as hays in a randomized block design. A 15-d adjustment period was followed by a 5-d collection period during which the hays were consumed ad libitum. Voluntary intake of DM was greater (P<.01) for alfalfa (10.9 kg/d) than for the mean of the grasses, and intake of Matua (10.0 kg/d) was greater (P<.001) than that of bermudagrass (7.4 kg/d). Apparent digestibility of OM was greater (P<.001) for alfalfa (74%) than for the mean of the grasses but did not differ between Matua (64%) and bermudagrass (60%). At the end of the digestion trial, each yearling was offered each of the three forage hays during an 11-d period to determine subsequent preference and effect of previous hay experience. Yearlings preferred alfalfa over the grass hays and generally selected more Matua than bermudagrass. All yearlings consumed less of the forage species to which they had been previously exposed compared with unadapted yearlings. The Matua hay fed in this trial was palatable and met most of the nutritional needs for yearling horses.

Map of cis-acting sequences that determine alternative pre-mRNA processing in the E3 complex transcription unit of adenovirus.

The E3 complex transcription unit of adenovirus encodes four major mRNAs (a, c, f, and h) and two minor (d and e) mRNAs with overlapping exons, alternative splice sites, and two polyadenylation sites, termed E3A (upstream) and E3B (downstream). mRNAs a and d use the E3A polyadenylation site, and mRNAs c, e, f, and h use the E3B site. We have analyzed virus mutants with deletions throughout the E3 region in order to identify cis-acting sequences that function in E3 pre-mRNA processing. The results presented in this report as well as previous results are summarized as follows. (i) Deletions in the first (5') intron at nucleotides (nt) 372 to 768 in E3 had no effect unless they removed the consensus sequence for the nt 372 5' splice site; however, the overall pattern of E3 mRNAs did not change significantly. (ii) Deletions in region I (nt 1441 to 2044) eliminated mRNAs a and c and resulted in corresponding increases in mRNAs f and h; we propose that region I contains sequences that suppress splicing. (iii) Mutations in region II (nt 2161 to 2243) resulted in nearly exclusive synthesis of mRNA f; this phenotype is understood and is discussed. (iv) Changing the AUUAAA component of the E3A poly(A) addition signal to AAUAAA resulted in increased mRNA a levels, suggesting that the E3A poly(A) addition signal is intrinsically inefficient. (v) Deletions in region III (nt 2488 to 3002) decreased mRNA a levels about two- to threefold and specifically increased mRNA f levels; we suggest that region III facilitates use of the E3A polyadenylation site. (vi) Deletions in region IV (nt 2904 to 3251) increased mRNA a levels about two- to threefold; we suggest that region IV may contain sequences that facilitate use of the E3B polyadenylation site. A map of sequences that determine alternative pre-mRNA processing in region E3 is now nearly complete.

Epidermal growth factor receptor is down-regulated by a 10,400 MW protein encoded by the E3 region of adenovirus.

Epidermal growth factor (EGF) binds to specific high affinity receptors (EGF-Rs) and induces endosome-specific internalization and degradation of ligand-receptor complexes in lysosomes. We report here that EGF-R is down-regulated in an analogous manner during early infection of a variety of cell types by group C human adenoviruses. This effect is not a function of viral entry, nor is it due to a nonspecific increase in turnover of membrane proteins. Using a series of virus deletion mutants, the gene responsible for EGF-R down-regulation was mapped to the E3 transcription unit. The E3 gene product, a protein of MW 10,400 (10.4K), induces internalization and degradation of EGF-R, but does not affect synthesis of the EGF-R precursor. The 10.4K protein is not an EGF-like autocrine growth factor, but is similar in sequence to a region in EGF-R at the cytoplasmic face of the transmembrane domain. This suggests that down-regulation of EGF-R during adenovirus infection may occur by a novel mechanism that involves the formation of hetero-oligomers composed of 10.4K and EGF-R.

Competition between splicing and polyadenylation reactions determines which adenovirus region E3 mRNAs are synthesized.

Complex transcription units encode multiple mRNAs which arise by alternative processing of a common pre-mRNA precursor. It is not known how the pre-mRNA processing pathways are determined or controlled. We are investigating this problem by using the E3 complex transcription unit of adenovirus as a model. Our approach is to construct virus mutants with lesions in E3 and then determine how the mutation affects the accumulation of E3 mRNAs in vivo. We report results which indicate that competition between splicing reactions and polyadenylation reactions occurs in vivo and that this plays an important role in alternative pre-mRNA processing.

A 14,700 MW protein from the E3 region of adenovirus inhibits cytolysis by tumor necrosis factor.

We find that cells infected with wild-type group C human adenoviruses are not killed by exposure to tumor necrosis factor (TNF), but cells infected with adenoviruses that delete the E3 transcription unit are highly sensitive to TNF lysis. Mock-infected cells are resistant to TNF. Thus, adenovirus infection induces cellular susceptibility to lysis by TNF, and a product of E3 protects against lysis by TNF. The E3-dependent resistance to TNF was investigated using virus mutants that delete different segments of E3. Resistance was found to depend on the presence of a 14,700 MW protein, which has only recently been identified and for which there was no known function. Our results support the hypothesis that one of the functions of TNF in vivo is to combat virus infections, and that the 14,700 MW protein evolved in adenovirus to counteract the antiviral effects of TNF.

Identification of a novel sequence that governs both polyadenylation and alternative splicing in region E3 of adenovirus.

Region E3 encodes four major overlapping mRNAs with different splicing patterns. There are two poly(A) sites, an upstream site called E3A and a downstream site called E3B. We have analyzed virus mutants with deletions or insertions in E3 in order to identify sequences that function in the alternative processing of E3 pre-mRNAs, and to understand what determines which poly(A) sites and which splice sites are used. In previous studies we established that the 5' boundary of the E3A poly(A) signal is at an ATTAAA sequence. We now show, using viable virus mutants, that the 3' boundary of the E3A signal is located within 47-62 nucleotides (nt) downstream of the ATTAAA (17-32 nt downstream of the last microheterogenous poly(A) addition site). Our data further suggest that the spacing between the ATTAAA, the cleavage sites, and the essential downstream sequences may be important in E3A 3' end formation. Of particular interest, these mutants suggest a novel mechanism for the control of alternative pre-mRNA processing. Mutants which are almost completely defective in E3A 3' end formation display greatly increased use of a 3' splice site located 4 nt upstream of the ATTAAA. The mRNA that uses this 3' splice site is polyadenylated at the E3B poly(A) site. We suggest, for this particular case, that alternative pre-mRNA processing could be determined by a competition between trans-acting factors that function in E3A 3' end formation or in splicing. These factors could compete for overlapping sequences in pre-mRNA.

Deletion mutants that alter differential RNA processing in the E3 complex transcription unit of adenovirus.

Region E3 of the adenovirus encodes about ten overlapping mRNAs (a to j) with different splicing patterns and with two RNA 3' end sites termed E3A and E3B. We have examined how deletions in 12 viable virus mutants affect differential RNA processing in E3. We assayed E3 mRNAs by the nuclease-gel and RNA blot procedures. Some deletions had no effect whereas others (e.g. deletion of a 3' splice or the E3A 3' end signal) had the anticipated effects on RNA processing. However, deletions in two regions had surprising effects. Deletions in one region (nucleotides 1691 to 2044) enhanced splicing at the upstream 951 5' splice site and the downstream 2157 and/or 2880 3' splice sites. Some of these deletions prevented RNA 3' end formation at the downstream E3A site. Deletion in the other region (nucleotides 2173 to 2237) enhanced an upstream splice site (951 to 2157) such that almost all pre-mRNA was processed into mRNA f. We suggest that these two regions contain cis-acting signals that regulate differential RNA processing. We discuss the results in terms of RNA folding and scanning models for splicing, as well as models for differential RNA 3' end formation at the E3A versus the E3B site.

Virus deletion mutants that affect a 3' splice site in the E3 transcription unit of adenovirus 2.

Five viable virus mutants were constructed with deletions near a 3' splice site located at nucleotide 2157 in the E3 transcription unit of adenovirus 2. The mutants were examined for splicing activity at the 2157 3' splice site in vivo by nuclease-gel analysis of steady-state cytoplasmic mRNA. Splicing was not prevented by an exon deletion (dl719) that leaves 16 5'-proximal exon nucleotides intact or by intron deletions that leave 34 (dl717, dl712) or 18 (dl716) 3'-proximal intron nucleotides intact. The sequences deleted in one of these intron mutants (dl716) include the putative branchpoint site used in lariat formation during splicing. Thus, a surrogate branchpoint site apparently can be used for splicing. Another intron mutant (dl714) has a deletion that leaves 15 3'-proximal intron nucleotides intact; remarkably, this deletion virtually abolished splicing, even though the deletion is only 3 nucleotides closer to the splice site than is the deletion in dl716 which splices normally. The three nucleotides deleted in dl714 that are retained by dl716 are the sequence TGT. The TGT sequence is located on the 5' boundary of the pyrimidine-rich region upstream of the nucleotide 2157 3' splice site. Such pyrimidine-rich regions are ubiquitous at 3' splice sites. Most likely, the TGT is required for splicing at the nucleotide 2157 3' splice site. The TGT may be important because of its specific sequence or because it forms the 5' boundary of the pyrimidine-rich region.

Caesarean section for eclampsia

The literature concerning Caesarean section for eclampsia has been reviewed. A small series of cases treated single-handed by an experienced practitioner in an isolated country hospital has been reviewed. The results recorded are encouraging. Some conclusions are drawn, leading to a suggestion that the method be given a further trial. (AU)