We report an improved draft nucleotide sequence of the 2.3-gigabase genome of maize, an important crop plant and model for biological research. Over 32,000 genes were predicted, of which 99.8% were placed on reference chromosomes. Nearly 85% of the genome is composed of hundreds of families of transposable elements, dispersed nonuniformly across the genome. These were responsible for the capture and amplification of numerous gene fragments and affect the composition, sizes, and positions of centromeres. We also report on the correlation of methylation-poor regions with Mu transposon insertions and recombination, and copy number variants with insertions and/or deletions, as well as how uneven gene losses between duplicated regions were involved in returning an ancient allotetraploid to a genetically diploid state. These analyses inform and set the stage for further investigations to improve our understanding of the domestication and agricultural improvements of maize.

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The peptides vasopressin (VP) and oxytocin are derived from preprohormone precursers encoded by highly homologous linked genes that are expressed in discrete groups of hypothalamic neurons. The mature hormones are released into the peripheral circulation from the neural (posterior) lobe of the pituitary and have also been implicated in the regulation of anterior lobe. We have used Northern blotting and in situ hybridization to RNA in tissue sections to describe the presence, anatomical localization, and regulation of VP and oxytocin RNAs in the pituitary gland itself. We were unable to detect VP transcripts in the anterior and intermediate lobes of the pituitary. Rather we found low levels of VP RNA in the neural lobe. Furthermore, the osmotic stimulation of a 2%

In rats, a shift from somatotroph dominance to lactotroph dominance during pregnancy and lactation is well reported. Somatotroph to lactotroph transdifferentiation and increased lactotroph mitotic activity are believed to account for this and associated pituitary hypertrophy. A combination of cell death and transdifferentiation away from the lactotroph phenotype has been reported to restore non-pregnant pituitary proportions after weaning. To attempt to confirm that a similar process occurs in mice, we generated and used a transgenic reporter mouse model (prolactin (PRL)-Cre/ROSA26-expression of yellow fluorescent protein (EYFP)) in which PRL promoter activity at any time resulted in permanent, stable, and highly specific EYFP. Triple immunochemistry for GH, PRL, and EYFP was used to quantify EYFP+ve, PRL−ve, and GH+ve cell populations during pregnancy and lactation, and for up to 3 weeks after weaning, and concurrent changes in cell size were estimated. At all stages, the EYFP reporter was expressed in 80% of the lactotrophs, but in fewer than 1% of other pituitary cell types, indicating that transdifferentiation from those lactotrophs where reporter expression was activated is extremely rare. Contrary to expectations, no increase in the lactotroph/somatotroph ratio was seen during pregnancy and lactation, whether assessed by immunochemistry for the reporter or PRL: findings confirmed by PRL immunochemistry in non-transgenic mice. Mammosomatotrophs were rarely encountered at the age group studied. Individual EYFP+ve cell volumes increased significantly by mid-lactation compared with virgin animals. This, in combination with a modest and non-cell type-specific estrogen-induced increase in mitotic activity, could account for pregnancy-induced changes in overall pituitary size.

We investigated the effects of thyroid status on nitric oxide synthase (NOS) gene expression in the rat hypothalamic paraventricular (PVN) and supraoptic nuclei (SON). Propylthiouracil (PTU)-induced hypothyroidism in male rats produced a highly significant reduction in NOS gene transcripts in the PVN and SON, as assessed by quantitative in situ hybridization histochemistry with a specific oligodeoxynucleotide probe. The addition of T3 (40 micrograms/kg) to the PTU-containing diet completely prevented the reduction in NOS transcripts. Hyperthyroidism, induced by adding 160 micrograms/kg T3 to the food, more than doubled the prevalence of NOS transcripts in the PVN and SON after a similar time. Up-regulation of NOS gene transcripts induced by the osmotic stimulus of chronic salt loading was markedly attenuated by PTU-induced hypothyroidism. These results demonstrate a major effect of thyroid status on regulation of NOS gene expression in the hypothalamus.

Dietary fat is ingested in three forms: 1) in solid food, 2) as aqueous emulsions, and 3) as unemulsified, liquid oil. On the basis of a scant previous literature, we postulated that liquid fat (emulsions or oils) would empty from the stomach at speeds that varied with the amounts ingested but that this dynamic would be modulated by feedback inhibition from lipolytic products. To test these ideas, we used a gamma camera to track gastric emptying of 123I-labeled fat in dogs with chronic pancreatic fistulas by which lipase was excluded from or replenished in the duodenum in varied amounts after dogs were fed 15-, 30-, and 60-g loads of liquid fat given with solid foods or as emulsions. We also tracked concurrent gastric emptying of 113mIn, which marked the solid food phase or the water phase of emulsions. In some studies, we used a potent and specific inhibitor (orlistat) of pancreatic and gastric lipases to assess how lipolytic products modulated emptying of liquid fat. In the absence of pancreatic enzymes, both oils and emulsions emptied initially at high speeds that varied with fat loads, but emptying slowed 20 min after ingestion of emulsions and 60 min after ingestion of unemulsified oil. Studies with orlistat indicated that these changes in rates resulted from liberation of gastric lipolytic products. Emptying of oil emulsions was not altered by duodenal replenishment with pancreatic enzymes, but emptying of unemulsified oil was inhibited in a dose-related fashion, such that maximal inhibition was achieved when pancreatic enzymes were replenished at > or = 40% of normal amounts. Studies with orlistat confirmed that this dose-dependent slowing was due specifically to lipase. Emptying of solid food was much more sensitive to replenishment with enzymes, so that a 10% replenishment maximally inhibited solid emptying.

Melanin-concentrating hormone (MCH) gene expression in the brain of rainbow trout, reared and maintained in either pale or black-coloured tanks, was studied using in situ hybridization histochemistry. MCH transcripts were most prevalent in the magnocellular neurones of the nucleus lateralis tuberis (NLT), which project to the pituitary gland. They were also present, although at much lower levels, in dorsally projecting parvocellular neurones, sited more posteriorly above the lateral ventricular recess (LVR). In the NLT the most intense hybridization signal was seen over the pituitary stalk; above the LVR, the most active neurones were located caudally. In both the NLT and above the LVR, MCH hybridization signal was 4-fold stronger in white-reared fish than in black-reared fish. There was also a marked diurnal variation in MCH expression in both sites, with high levels at 16.00 h and lower levels at 04.00 h. The results show that gene activity in both hormonal (NLT) and neuromodulator/neurotransmitter (LVR) MCH neurones is induced by pale environmental colour and that MCH gene activity is subject to pronounced diurnal variation.

INTRODUCTION Some patients report continuing pain and functional limitations after total knee replacement (TKR). While numbness around the TKR scar is common, the impact of numbness is less clear. One particular activity that could be influenced by numbness is kneeling. The aim of this study was to explore the impact of numbness around TKR scars on health related quality of life and kneeling ability. METHODS Fifty-six patients were recruited one year after primary TKR. Sensation around the knee was assessed through patient self-reporting, monofilament testing and vibration, and patients' distress was measured on a visual analogue scale. Patient reported outcome measures (PROMs) including the Western Ontario and McMaster Universities (WOMAC) index, the Knee injury and Osteoarthritis Outcome Score (KOOS), the painDETECT (Pfizer, Berlin, Germany) questionnaire and the EQ-5D™ (EuroQol, Rotterdam, Netherlands) questionnaire were used. Participants were also asked about kneeling ability. RESULTS While 68% of patients reported numbness around their TKR scar, there was no statistically significant correlation between numbness and distress at numbness (self-report: 0.23, p=0.08; monofilament: 0.15, p=0.27). Furthermore, numbness did not correlate significantly with joint specific PROMs (WOMAC: 0.21, p=0.13; KOOS: 0.18, p=0.19). However, difficulty with kneeling did correlate with both self-reported numbness (0.36, p=0.020) and worse PROM scores (WOMAC pain subscale: 0.62, p<0.001; KOOS: 0.64, p<0.001). CONCLUSIONS Numbness after knee replacement is common but is not associated with worse patient reported outcomes.