Fertility and genomics: comparison of gene expression in contrasting reproductive tissues of female cattle.

To compare gene expression among bovine tissues, large bovine RNA-seq datasets were used, comprising 280 samples from 10 different bovine tissues (uterine endometrium, granulosa cells, theca cells, cervix, embryos, leucocytes, liver, hypothalamus, pituitary, muscle) and generating 260 Gbases of data. Twin approaches were used: an information-theoretic analysis of the existing annotated transcriptome to identify the most tissue-specific genes and a de-novo transcriptome annotation to evaluate general features of the transcription landscape. Expression was detected for 97% of the Ensembl transcriptome with at least one read in one sample and between 28% and 66% at a level of 10 tags per million (TPM) or greater in individual tissues. Over 95% of genes exhibited some level of tissue-specific gene expression. This was mostly due to different levels of expression in different tissues rather than exclusive expression in a single tissue. Less than 1% of annotated genes exhibited a highly restricted tissue-specific expression profile and approximately 2% exhibited classic housekeeping profiles. In conclusion, it is the combined effects of the variable expression of large numbers of genes (73%-93% of the genome) and the specific expression of a small number of genes (<1% of the transcriptome) that contribute to determining the outcome of the function of individual tissues.

Glycoproteins and glycosidases of the cervix during the periestrous period in cattle.

The cervix and its secretions undergo biochemical and physical changes under the differential influences of estrogen and progesterone. These include changes in the glycoprotein profile of the endocervix and its secretions. A comprehensive survey of such changes in cervical epithelium and cervical secretions was performed on bovine samples throughout the periestrous period. Cervical tissue samples and swabs were collected from synchronized beef heifers that were slaughtered 1) 12 h after controlled intravaginal progesterone-releasing device (CIDR) removal, 2) 24 h after CIDR removal, 3) at the onset of estrus, 4) 12 h after the onset of estrus, 5) 48 h after the onset of estrus, and 6) 7 d after the onset of estrus. Histological staining with hematoxylin and eosin, periodic acid Schiff, Alcian blue, and high-iron diamine was carried out to map overall patterns of stored glycoproteins and tissue structure. Biotinylated lectins were also used to detect the presence and distribution of a range of saccharide structures. The activities of ß-galactosidase, α-L-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase were measured in cervical swabs using specific substrates. The epithelial layer of the cervix exhibited dynamic changes in cellular hypertrophy and amounts of stored glycoprotein. The greatest content of neutral and acidic mucins was observed 48 h after onset of estrus (P < 0.05). Sialylated mucins predominated at the bases of cervical folds, whereas sulfated mucins were more abundant (P < 0.05) at their apices. The stained area of core mucin glycans changed (P < 0.05) in association with follicular versus luteal phases, whereas terminal glycans changed (P < 0.05) mainly at the time of estrus and shortly thereafter. The greatest activity of ß-galactosidase and sialidase was observed 12 h after onset of estrus, whereas ß-hexosaminidase and α-fucosidase peaked at the luteal time point (P < 0.05). Taken together, we suggest that the well-known changes in the endocervix and its secretions that are associated with the physiological modulation of sperm transport and function of the cervical barrier are, in part, driven by glycosylation changes.

Coronary arteries of the roe deer (Capreolus capreolus; Linnaeus 1758) heart.

A study of the coronary arteries of the roe deer heart was performed on 21 hearts of animals of both sexes and various ages. The roe deer heart is supplied by two arteries: the left coronary artery and the right coronary artery. The left coronary artery arises from the left aortic sinus and forms a short common trunk. The left coronary artery reaches the coronary groove, then divides into the paraconal interventricular branch and the circumflex branch. The circumflex branch gives off several branches to the left ventricle wall and terminates in the subsinuosal interventricular groove as the subsinuosal interventricular branch. The right coronary artery is less pronounced than the left coronary artery. It arises from the right aortic sinus and enters the coronary groove as the right circumflex branch. We found the left arterial cone branch in 75% and the right arterial cone branch in 80% of the cases investigated. The coronary arteries of the heart run subepicardially. In 9 cases we found muscular bridges over the coronary arteries, mostly on the paraconal interventricular branch. In conclusion we affirm the left type of the arterial vascularisation in the roe deer heart.

Capsule endoscopy: a single-centre experience with the first 226 capsules.

BACKGROUND: Capsule endoscopy (CE) refers to a novel diagnostic method of imaging the gastrointestinal tract using a wireless capsule that transmits images to a data recorder while the device traverses the small intestine. OBJECTIVE: To review the authors' experience with CE to determine the indications, outcomes and management of positive findings. METHODS: Patients were prepared for CE with a single dose of magnesium citrate. Following an 8 h fast, a sensor array system was applied to the abdomen, the capsule was swallowed and the images were transmitted to a data recorder worn on the patient's side. Typically, the battery life of the capsule is 8 h, following which the data recorder is returned, downloaded to a computer workstation and reviewed. RESULTS: To date, 226 capsule studies have been performed in 209 patients. The indications included obscure bleeding (167 studies: 88 overt, 79 occult), anemia (14 studies), evaluation for inflammatory bowel disease (12 studies), screening for polyps (10 studies), pain (19 studies) and abnormal radiological imaging (4 studies). In the setting of obscure bleeding, a definitive source of bleeding was discovered in 85 studies. This included angiodysplasia (52 studies), mitotic lesions (10 studies) and ulcers (23 studies). A probable source of bleeding was found in another 10 capsule studies. In the setting of anemia without evidence of bleeding, the definitive findings included ulcers (three studies), angiodysplasia (two studies), mitotic lesions (one study) and celiac disease (one study). Of four patients with abnormal radiological imaging, CE demonstrated lesions in two. The results of 35 capsule studies led to laparotomy with curative surgical resection. In eight studies, the capsules became lodged within a stricture; none led to obstruction and three were managed endoscopically. CONCLUSION: The yield of CE in carefully selected patients with obscure bleeding approximates 51%. There appear to be few complications, and patient satisfaction appears high. Cost analysis and further studies of clinical outcomes are required to elucidate appropriate indications for this device.

Maf1p, a negative effector of RNA polymerase III in Saccharomyces cerevisiae.

Although yeast RNA polymerase III (Pol III) and the auxiliary factors TFIIIC and TFIIIB are well characterized, the mechanisms of class III gene regulation are poorly understood. Previous studies identified MAF1, a gene that affects tRNA suppressor efficiency and interacts genetically with Pol III. We show here that tRNA levels are elevated in maf1 mutant cells. In keeping with the higher levels of tRNA observed in vivo, the in vitro rate of Pol III RNA synthesis is significantly increased in maf1 cell extracts. Mutations in the RPC160 gene encoding the largest subunit of Pol III which reduce tRNA levels were identified as suppressors of the maf1 growth defect. Interestingly, Maf1p is located in the nucleus and coimmunopurifies with epitope-tagged RNA Pol III. These results indicate that Maf1p acts as a negative effector of Pol III synthesis. This potential regulator of Pol III transcription is likely conserved since orthologs of Maf1p are present in other eukaryotes, including humans.

Suppressors of translation initiation defect in hem12 locus of Saccharomyces cerevisiae.

A system for the positive selection of transational initiation suppressors in S. cerevisiae has been developed. A mutant with an ATA initiation codon in the HEM12 gene, encoding uroporphyrinogen decarboxylase, was used to select cis- and trans-acting suppressors. These suppressors partially restore growth on nonfermentable carbon sources, such as glycerol, but still allow the accumulation of porphyrins. All extragenic suppressors are mapped to the SUI1 locus, encoding initiation factor eIF1. The effect of the hem12 mutation is also partially reversed by the known SUI3 suppressor encoding the beta subunit of eIF2. In contrast, the sui2 suppressor encoding the a subunit of eIF2 does not affect the hem12 phenotype. The intragenic suppressors are able to restore the translation of hem12 due to the generation of additional, in frame AUG codons upstream of the hem12-14 mutation. Mutational analysis of the HEM12 leader sequence was also performed to determine the role of small open reading frames (uORFs) present upstream of the HEM12 ORF. Studies on the expression of integrated hem12-1/4-lacZ fusion, devoid of all upstream ATGs, indicate a lack of regulatory effect of uORFs on HEM12 translation.

Infectious transcripts from cloned cDNA of potato leafroll luteovirus.

Infectious transcripts play a key role in the research on plant viruses at the molecular level. A number of cDNA clones covering the whole genome of the Polish isolate of potato leafroll virus were constructed. Four overlapping clones were selected and assembled using restriction sites. The full copy was positioned between T7 RNA polymerase promoter and unique ScaI site. The full-length capped transcripts of the sequence of the viral genome synthesised in vitro were able to replicate in protoplasts and to produce the viral coat protein.