Herbage intake and milk production of late-lactation dairy cows offered a second-year chicory crop during summer.

Chicory (Cichorum intybus L.) is a summer-active forage herb which has been proposed as an option to increase summer feed supply, increase dry matter intake, nutrient intake, and milk yield from nonirrigated dairy production systems in southern Australia. Dry matter intake, nutrient intake, milk yield, and yield of milk fat and protein of predominantly Holstein-Friesian dairy cows in late lactation consuming 3 herbage-based diets (4 replicates per treatment) were measured. The 3 grazed herbages were second-year chicory (CHIC) and perennial ryegrass (Lolium perenne L.; PRG) monocultures and a mixed sward (~50:50) of chicory and perennial ryegrass (MIX). All diets (CHIC, PRG, and MIX) were supplemented with alfalfa (Medicago sativa L.) hay (5.5kg of DM/cow per day) and an energy-based concentrate pellet (4.0kg of DM/cow per day). There were no significant differences in milk yield (12.0 to 12.6kg/d across the treatments) or the yield of milk fat (539 to 585g/d) and milk protein (433 to 447g/d) between the 3 herbage-based diets. No differences in DMI (17.9 to 19.2kg/d) or estimated metabolizable energy intake (173 to 185MJ/d) were noted between treatments. Estimated metabolizable energy concentrations in the forages on offer were lower in CHIC than PRG (7.6 vs. 8.2MJ/kg of dry matter), but the concentration in consumed herbage was not different (9.1 vs. 9.2MJ/kg of dry matter); as such, potential for increased milk yield in cows offered CHIC was limited. Increased concentration of polyunsaturated fatty acids was observed in chicory herbage compared with perennial ryegrass. This was associated with increased milk conjugated linoleic acid and milk polyunsaturated fatty acids when chicory formed part of the diet (CHIC compared to PRG and MIX). Chicory could be used as an alternative to perennial ryegrass in summer; however, the developmental stage of chicory will influence concentrations of metabolizable energy and neutral detergent fiber and, therefore, intake and milk production responses compared with perennial ryegrass.

Milk production and composition of mid-lactation cows consuming perennial ryegrass-and chicory-based diets.

Dry matter intakes (DMI), nutrient selection, and milk production responses of dairy cows grazing 3 herbage-based diets offered at 2 allowances were measured. The 2 allowances were 20 (low) and 30 (high) kg of dry matter (DM)/cow per day and these were applied to 3 herbage types: perennial ryegrass (PRG) and chicory (CHIC+) monocultures and a mixed sward of chicory and perennial ryegrass (MIX). The CHIC+ diet was supplemented with alfalfa hay (approximately 2 kg of DM/cow per day) to maintain dietary neutral detergent fiber (NDF) concentration and all diets were supplemented with energy-based pellets (6 kg of DM/cow per day). Holstein-Friesian dairy cows averaging 136 ± 30 d in milk were allocated to 4 replicates of the 6 treatments using stratified randomization procedures. Cows were adapted to their experimental diets over a 14-d period, with measurements of DMI, milk yield, and composition conducted over the following 10 d. Herbage DMI was lowest (12.8 vs. 14.0 kg of DM/d) for CHIC+ compared with the MIX and PRG, although total forage intake (grazed herbage plus hay) was similar (14.0 to 15.0 kg of DM/d) across the 3 treatments. Milk production, milk protein, and milk fat concentrations were not different between herbage types. Grazed herbage DMI increased with increasing herbage allowance and this was associated with increased milk protein concentration (3.23 to 3.34%) and total casein production (41.7 to 43.6 mg/g). Concentrations of polyunsaturated fatty acids in milk fat, particularly linoleic acid, were increased in milk from cows offered the CHIC+ or the MIX diets, indicating potential benefits of chicory herbage on milk fatty acid concentrations. Although feeding CHIC+ or MIX did not increase milk yield, these herbage types could be used as an alternative to perennial ryegrass pasture in spring.

Interobserver reliability of the radiographic assessment of cement fixation in total hip arthroplasty.

BACKGROUND: The objective of this work is to evaluate the performance of a modified quantitative Barrack Cement grading in primary THA. Previous studies demonstrated both poor intraobserver and interobserver reliability which may be due to the qualitative nature of the scale. METHODS: Interobserver reliability of the Barrack Cement Grading System in its original format and then in combination with a quantitative measurement of implant/cement lengths was evaluated on 50 immediate post-operative radiographs of primary cemented arthroplasties. Intraobserver reliability was also assessed on a sub-sample of radiographs. Three evaluators with different skill levels and specialty participated: an arthroplasty surgeon, an orthopaedic resident and a radiologist. Reliability was measured using a weighted kappa coefficient for paired comparisons among the evaluators. RESULTS: Interobserver reliability was poor (κ < 0.10) for all pairings of the Barrack System. The modified quantitative system achieved slight (κ < 0.20) to poor reliability. Intraobserver reliability was dependent on the skill and specialty of the evaluator with maximal values achieved for the experienced arthroplasty surgeon using the modified quantitative system (κ = 0.62). CONCLUSION: Use of the modified scale may improve the reliability of ratings when used by individual experienced arthroplasty surgeons.

Release of Ca2+ from individual plant vacuoles by both InsP3 and cyclic ADP-ribose.

Calcium mobilization from intracellular pools couples many stimuli to responses in plant cells. Cyclic adenosine 5'-diphosphoribose (cADPR), which interacts with a ryanodine receptor in certain animal cells, was shown to elicit calcium release at the vacuolar membrane of beet storage root. The vacuolar calcium release pathway showed similarities to cADPR-gated calcium release in animal cells, including inhibition by ruthenium red, ryanodine activation, and high affinity for cADPR [Michaelis constant (Km) = 24 +/- 7 nanomolar]. Analysis by patch-clamping demonstrated that the cADPR-gated pathway in beet is voltage-dependent over the physiological range, does not spontaneously desensitize, and is colocalized with an inositol 1,4,5-trisphosphate (InsP3)-gated calcium release pathway in individual vacuoles.

Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols.

Tomatoes are an excellent source of the carotenoid lycopene, a compound that is thought to be protective against prostate cancer. They also contain small amounts of flavonoids in their peel ( approximately 5-10 mg/kg fresh weight), mainly naringenin chalcone and the flavonol rutin, a quercetin glycoside. Flavonols are very potent antioxidants, and an increasing body of epidemiological data suggests that high flavonoid intake is correlated with a decreased risk for cardiovascular disease. We have upregulated flavonol biosynthesis in the tomato in order to generate fruit with increased antioxidant capacity and a wider range of potential health benefit properties. This involved transformation of tomato with the Petunia chi-a gene encoding chalcone isomerase. Resulting transgenic tomato lines produced an increase of up to 78 fold in fruit peel flavonols, mainly due to an accumulation of rutin. No gross phenotypical differences were observed between high-flavonol transgenic and control lines. The phenotype segregated with the transgene and demonstrated a stable inheritance pattern over four subsequent generations tested thus far. Whole-fruit flavonol levels in the best of these lines are similar to those found in onions, a crop with naturally high levels of flavonol compounds. Processing of high-flavonol tomatoes demonstrated that 65% of flavonols present in the fresh fruit were retained in the processed paste, supporting their potential as raw materials for tomato-based functional food products.

Amplification, increased dosage and in situ expression of the telomerase RNA gene in human cancer.

Telomere length is maintained by the enzyme, telomerase, which has been linked to cellular immortality and tumour progression. However, the reasons for the high levels of telomerase found in human tumours are unknown. We have mapped the human telomerase RNA gene, (hTR), to chromosome 3q26.3 and show the hTR gene to be amplified in four carcinomas, (2/33 cervix, 1/31 head and neck, 1/9 lung). In addition, increased copy numbers of the hTR locus was also observed in 97% of tumours. By in situ hybridisation, the histological distribution of high levels of hTR expression could be demonstrated in a lung tumour and its metastasis with hTR amplification. These results are the first report of genetic alterations involving a known component of telomerase in human cancer. Indeed, it is also the first report of the amplification of a specific locus within the chromosome 3q region frequently subject to copy number gains in human tumours. In addition, we also show for the first time the histological distribution of the RNA component of telomerase in human tumours.

Cloning and characterization of human and mouse telomerase RNA gene promoter sequences.

Variation in telomerase activity is correlated with cellular senescence and tumour progression. However, although the enzymatic activity of telomerase has been well studied, very little is known about how expression of telomerase genes is regulated in mammalian cells. We have therefore cloned the promoter regions of the human (hTR), and mouse, (terc), telomerase RNA genes in order to identify the regulatory elements controlling telomerase RNA gene transcription. 1.76 kb encompassing the hTR gene promoter region was sequenced, as was 4 kb encompassing the terc promoter. No significant sequence similarity could be detected in comparisons between human and mouse 5'-regions, flanking the transcribed sequences. However, both the human and mouse telomerase RNA genes are within CpG islands and may therefore be under the regulation of DNA methylation. Transient expression of hTR-reporter gene constructs in HeLa and GM847 cells identified the elements responsible for promoter activity are contained in a 231 bp region upstream of the transcriptional start site. Transient expression of terc-reporter gene constructs in Swiss3T3 and A9 cells identified the elements responsible for promoter activity are contained in a 73 bp region upstream of the transcriptional start site. These studies have implications for novel transcription targeted cancer therapies.

Light-weight lead aprons--light on weight, protection or labelling accuracy?

X-ray transmission tests were performed on a Green-Lite (Infab Corporation) apron/vest combination, and compared to a number of other apron/vest combinations routinely used at Christchurch Hospital as well as a sheet of 0.5 mm lead. The materials were X-rayed using the primary beam of a Philips Optimus 50 X-ray machine over an energy range of 50-125 kVp. The entrance and exit doses were recorded and percentage transmission calculated for each kVp. The Green-Lite apron/vest (labelled as 0.5 mm lead at 85 kVp) relies on the overlap at the front to provide the nominal 0.5 mm protection for both the vest and the apron. It performed significantly worse than 0.5 mm of lead and other 0.5 mm lead equivalent apron/vest combinations and provided between 0.3 and 0.39 mm lead equivalent protection depending on the energy used. Vests from other manufacturers all achieved 0.5 mm lead equivalence for a single layer of vest material over the range of energies tested and so were comparable to 1.0 mm lead when doubled. Some aprons relied on a double layer of material to achieve the 0.5 mm lead equivalence (which was not always made clear on the label), while others stated their lead equivalence for a single layer. This resulted in some confusion among wearers of the aprons as to which apron was better.

Mapping of the gene for the human telomerase reverse transcriptase, hTERT, to chromosome 5p15.33 by fluorescence in situ hybridization.

Telomerase, the enzyme that maintains the ends of chromosomes, is absent from the majority of somatic cells but is present and active in most tumours. The gene for the reverse transcriptase component of telomerase (hTERT) has recently been identified. A cDNA clone of this gene was used as a probe to identify three genomic bacterial artificial chromosome (BAC) clones, one of which was used as a probe to map hTERT by fluorescence in situ hybridization (FISH) to chromosome 5p15.33. This BAC probe was further used to look at copy number of the hTERT region in immortal cell lines. We found that 10/15 immortal cell lines had a modal copy number of 3 or more per cell, with one cell line (CaSki) having a modal copy number of 11. This suggests that increases in copy number of the hTERT gene region do occur, and may well be one route to upregulating telomerase levels in tumour cells. 5p15 gains and amplifications have been documented for various tumour types, including non-small cell lung carcinoma, squamous cell carcinoma of head and neck, and uterine cervix cancer, making hTERT a potential target.

Pharmacology of Ca2+ release from red beet microsomes suggests the presence of ryanodine receptor homologs in higher plants.

Cyclic ADP-ribose (cADPR) is known to release Ca2+ from plant vacuoles, implying that this NAD+ metabolite may possess a second messenger role in plants. The degree to which the plant cADPR-gated Ca2+ release mechanism resembles cADPR action in animals has been evaluated. cADPR-elicited Ca2+ release from red beet microsomes was inhibited by 1 mM procaine but insensitive to heparin. Furthermore, pre-release of Ca2+ from red beet vesicles by either 5 mM caffeine or micromolar levels of ryanodine precluded further Ca2+ mobilisation by cADPR. Thus, this study argues strongly for conservation between the plant and animal cADPR-elicited Ca2+ release mechanisms.

Detection of cytomegalovirus in upper gastrointestinal biopsies from heart transplant recipients: comparison of light microscopy, immunocytochemistry, in situ hybridisation, and nested PCR.

AIM: To establish the diagnostic value of in situ hybridisation and the nested polymerase chain reaction (PCR) in detecting clinically relevant cytomegalovirus (CMV) infection in upper gastrointestinal biopsies from heart transplant patients. METHODS: Test sensitivity and specificity for detection of CMV early gene RNA by in situ hybridisation and CMV intermediate early gene by PCR were established and then compared with haematoxylin and eosin (H&E) and immunocytochemical detection of CMV in order to establish the best pathological diagnostic approach. All investigations were carried out on formalin fixed, paraffin embedded tissue. RESULTS: Nested PCR had the highest test sensitivity, followed by in situ hybridisation and immunocytochemistry with the same sensitivity; H&E had the lowest. H&E and immunocytochemistry were the most specific but both had a significant false negative rate which was less of a problem with PCR. However, PCR gave no other diagnostic information, and in situ hybridisation was no better than immunocytochemistry. Both in situ hybridisation and PCR were technically complex and more expensive. CONCLUSIONS: H&E and immunocytochemistry represent the best initial screen for CMV and other diseases in upper gastrointestinal biopsies from heart transplant patients. If H&E and immunocytochemistry were negative, nested PCR could significantly increase the diagnostic yield of clinically relevant CMV infection. In situ hybridisation appeared to have no advantages and some drawbacks compared with immunocytochemistry and PCR.

Friendship clique and peer influences on body image concerns, dietary restraint, extreme weight-loss behaviors, and binge eating in adolescent girls.

This study explored friendship variables in relation to body image, dietary restraint, extreme weight-loss behaviors (EWEBs), and binge eating in adolescent girls. From 523 girls, 79 friendship cliques were identified using social network analysis. Participants completed questionnaires that assessed body image concerns, eating, friendship relations, and psychological family, and media variables. Similarity was greater for within than for between friendship cliques for body image concerns, dietary restraint, and EWLBs, but not for binge eating. Cliques high in body image concerns and dieting manifested these concerns in ways consistent with a high weight/shape-preoccupied subculture. Friendship attitudes contributed significantly to the prediction of individual body image concern and eating behaviors. Use of EWLBs by friends predicted an individual's own level of use.

Measurement and analysis of single and multiple finger tapping in normal and Parkinsonian subjects.

A four-finger tapping sensor and associated software have been developed for the investigation of timing and rhythm performance and mechanisms in normal subjects and their disruption in neurological disorders. The tapping sensor comprises four electronic touch pads and pacing lights. A personal computer (PC) is used to control visual and auditory pacing, record the time and pad of each tap, and carry out several performance analyses including graphing, phase-space plots, calculation of spectra and autocorrelations, filtering and descriptive statistical analysis. A study was conducted to investigate disruptions of timing and rhythm in subjects with Parkinson's disease (PD). Seven Parkinsonian and ten control subjects undertook paced and unpaced finger tapping tests. The hastening phenomenon-tapping asynchronously at a speed faster than the pacing-was seen with similar occurrence in both normal and PD subjects and appears to be due to perceptual difficulties. No evidence was seen of an increased variability of tapping at particular frequencies, contrary to previous reports. Festinated tapping, in which subjects cycled between acceleration to near-maximum speed and abrupt slowing down, was seen only in PD subjects. As noneof these subjects showed significant hastening, it appears that hastening and festination are unrelated phenomena. Inspection of variations of finger tapping intervals gave no evidence for the presence of deterministic chaos in the control of rhythmic tapping. In speed tests performed with and without a weight attached to the finger, it was found that, for reasons which remain unclear, normal subjects increased their maximum tapping speed with the weight attached.

Why do adolescent girls watch their weight? An interview study examining sociocultural pressures to be thin.

Body concerns, dieting, and weight watching were examined in 30 year 10 adolescent girls. Semistructured interviews consisting of open-ended and rated questions assessed descriptions of and reasons for weight loss attempts, with an emphasis on noting sociocultural influences. Audiotaped and transcribed interviews were assessed for themes, coded and rated. Findings suggested a strong role of sociocultural influences leading to both unhealthy and healthy body attitudes and eating behaviors. Media and fashion were reported to exert the strongest pressures to be thin for subjects. While a few subjects reported direct pressures to diet from friends and parents, indirect social influences were more common. These influences included social comparison, joint dieting and avoidance of social disapproval.

Hepatic and renal tubular cell nuclear glycogenosis in goats.

Nuclear glycogenosis has not been reported in goats. This paper describes five cases of hepatocellular and/or renal tubular cell nuclear glycogenosis in goats in which the histological and ultrastructural details were studied. Our findings support the concept that nuclear glycogenosis reflects a compensatory adaptation by the cells rather than a degenerative process even though the initiating stimulus and pathogenesis are not known.

Five-year multicenter retrospective review of cyclobenzaprine toxicity.

Cyclobenzaprine (CBP) has a cyclic structure similar to amitriptyline. In overdose, CBP has been suggested to produce the cardiovascular and neurologic toxicity found with the cyclic antidepressants. To examine this possibility, a retrospective chart review of all cases of CBP exposure reported to five regional poison centers was performed for the years 1989-93. There were a total of 750 charts identified for CBP exposure, of which 523 had data sufficient for evaluation. There were 121 polydrug ingestions leaving 402 pure CBP ingestions. Ages ranged from 7 mo to 77 yrs, with a mean of 20 yrs; 26% were 6 yrs or less. Females comprised 63% of the patient group. No deaths occurred. Dysrhythmias beyond sinus tachycardia were infrequent, and none were life-threatening. No seizures occurred. Common effects were lethargy, sinus tachycardia, and agitation, and both hypertension and hypotension were seen. All symptomatic cases with a known time of ingestion were symptomatic within 4 h of ingestion. Doses ingested ranged from 5-1000 mg, with a mean of 133 mg. Asymptomatic and symptomatic patients had a mean dose ingested of 45 mg and 183 mg, respectively. Treatment was primarily gastrointestinal (GI) decontamination and supportive care. Other therapies required were mechanical ventilation, dopamine, fluid bolus, sedation, and foley catheter. Symptoms requiring treatment beyond GI decontamination did not occur with ingestions less than 100 mg. In conclusion, cyclobenzaprine does not appear to produce the life-threatening cardiovascular or neurologic effects of the cyclic antidepressants in doses less than 1 g. Lethargy and anticholinergic effects are prominent, though serious toxicity is infrequent.

Analysis of N,N-diethyl-m-toluamide (DEET) in human postmortem specimens.

N,N-diethyl-m-toluamide (DEET) levels in postmortem specimens of stomach and contents, blood, liver, and urine are reported following ingestion of the compound. DEET was analyzed by gas chromatography with an OV-101 column and a nitrogen phosphorus detector. The presence of the compound in the four postmortem specimens was confirmed by mass spectrometry.

Coccidioidomycosis in the llama: case report and epidemiologic survey.

An 8-year-old nongravid female llama with a 1-month history of progressive posterior paresis was referred because of suspected degenerative myelopathy secondary to copper deficiency or plant poisoning. Neurologic examination revealed loss of conscious proprioception and slightly depressed bilateral patellar reflexes. Electromyographic examination of hindlimb flexors and extensors did not elicit evidence of lower motor neuron disease. Possible fragmentation and mottling of the 10th thoracic vertebral body were noted radiographically. Results of a lumbar CSF tap, complete blood count, and fecal flotation were not diagnostic. In the face of poor prognosis, the llama was euthanatized. Postmortem and histologic evaluation revealed, in addition to disseminated visceral granulomas, an extradural pyogranulomatous mass compressing the cord laterally at the level of T-10. Large numbers of Coccidioides immitis were dispersed throughout the granulomas. Complement fixing antibody tests in 11 other herd members showed evidence of C immitis infection in three.

EMMA--an eye movement measurement and analysis system.

A system has been developed for stimulation, recording and analysis of a wide range of eye movements. Eye movements are stimulated with an LED bar or a video projector under the control of a PC. The eye movements are measured using a scleral reflection technique (IRIS instrument), and sampled and stored on a PC. A range of tests have been developed to measure saccadic and smooth pursuit eye movements. A variety of tools have been developed to assist in the analysis of the data. Several research studies have ably demonstrated the utility and versatility of the system.