Ruminal ergovaline and volatile fatty acid dynamics: Association with poor performance and a key growth regulator in steers grazing toxic tall fescue.

Fescue toxicosis (FT) is produced by an ergot alkaloid (i.e., ergovaline [EV])-producing fungus residing in toxic fescue plants. Associations between EV, decreased weight gain and ruminal volatile fatty acids are unclear. Feces, rumen fluid, and blood were collected from 12 steers that grazed non-toxic (NT) or toxic (E +) fescue for 28 days. The E + group exhibited decreased propionate (P), increased acetate (A), and increased ruminal A:P ratio, with similar trends in feces. Plasma GASP-1 (G-Protein-Coupled-Receptor-Associated-Sorting-Protein), a myostatin inhibitor, decreased (day 14) only in E + steers. Ergovaline was present only in E + ruminal fluid and peaked on day 14. The lower ruminal propionate and higher A:P ratio might contribute to FT while reduced GASP-1 might be a new mechanism linked to E + -related weight gain reduction. Day 14 ergovaline zenith likely reflects ruminal adaptations favoring EV breakdown and its presence only in rumen points to local, rather than systemic effects.

Bile acid aspiration associated with lung chemical profile linked to other biomarkers of injury after lung transplantation.

Aspiration of gastrointestinal contents has been linked to worse outcomes following lung transplantation but uncertainty exists about underlying mechanisms. We applied high-resolution metabolomics of bronchoalveolar lavage fluid (BALF) in patients with episodic aspiration (defined by bile acids in the BALF) to identify potential metabolic changes associated with aspiration. Paired samples, one with bile acids and another without, from 29 stable lung transplant patients were studied. Liquid chromatography coupled to high-resolution mass spectroscopy was used to interrogate metabolomic contents of these samples. Data were obtained for 7068 ions representing intermediary metabolites, environmental agents and chemicals associated with microbial colonization. A substantial number (2302) differed between bile acid positive and negative samples when analyzed by false discovery rate at q = 0.01. These included pathways associated with microbial metabolism. Hierarchical cluster analysis defined clusters of chemicals associated with bile acid aspiration that were correlated to previously reported biomarkers of lung injury including T cell granzyme B level and the chemoattractants CXCL9 and CXCL10. These data specifically link bile acids presence in lung allografts to inflammatory pathways known to segregate with worsening allograft outcome, and provide additional mechanistic insight into the association between reflux and lung allograft injury.

Lamb boning -- an occupational cause of carpal tunnel syndrome?

Whether an occupation can cause carpal tunnel syndrome requiring carpal tunnel decompression (CTD) is contentious. We compared the demographics and incidence rates in lamb-freezing workers with the general population who had CTD. In the general population there were 1002 (63%) females and 583 (37%) males, mean age 48 years, and the rate of CTD was 1.36/1000 per annum. In lamb-freezing workers there were 225 males (mean age 38.4 years) and 60 females (mean age 44.6 years); most workers required CTD in their first three seasons. Compared with the general population, the incidence rate ratios in all freezing workers was 16.8; boners, 51.6; meat packers, 22.8; and slaughtermen, 5.4. All groups had a greater rate of CTD than the general population. This study suggests that carpal tunnel syndrome can be directly caused by an occupation.

Colonic motor dysfunction in human diabetes is associated with enteric neuronal loss and increased oxidative stress.

BACKGROUND: Gastrointestinal dysfunction is very common in diabetic patients. We assessed the changes in the colonic enteric nervous system using colectomy specimens and intestinal biopsies from diabetic subjects and age-matched controls. METHODS: In control and diabetic colons, we determined the total ganglion area (hematoxylin-eosin staining), changes in neuronal markers-protein gene product 9.5, peripherin, neuronal nitric oxide synthase (nNOS), neuropeptide Y (NPY), choline acetyl transferase (ChAT) and vasoactive intestinal peptide (by immunostaining), apoptosis (cleaved caspase-3 staining) and reduced glutathione levels. Superoxide dismutase mRNA was determined in enteric ganglia isolated by laser capture micro dissection. Isometric muscle recording was used to assess contraction and relaxation responses of colonic circular muscle strips. Apoptosis in enteric neurons under hyperglycemia in vitro was determined by cleaved caspase-3 Western blotting and protective effects of lipoic acid were evaluated. KEY RESULTS: Diabetic subjects had higher incidence of lower gastrointestinal symptoms like constipation and diarrhea at baseline prior to surgery. Diabetic ganglia displayed significant decrease in ganglion size due to enhanced apoptosis and loss of peripherin, nNOS, NPY, and ChAT neurons. Reduced glutathione levels in the diabetic colon (HbA1C > 7%) were significantly less than the control, indicating increased oxidative stress. Colonic circular muscle strips from diabetic subjects showed impaired contraction and relaxation responses compared with the healthy controls. Hyperglycemia-induced cleaved caspase-3 in enteric neurons was reversed by lipoic acid. CONCLUSIONS & INFERENCES: Our data demonstrate loss of enteric neurons in the colon due to increased oxidative stress and apoptosis which may cause the motility disturbances seen in human diabetes. Antioxidants may be of therapeutic value for preventing motility disorders in diabetes.

Changes in chromatin fiber density as a marker for pluripotency.

Extensive alterations in chromatin structure at the nucleosome level are linked to developmental potential. We hypothesize that such alterations in chromatin structure reflect and, to some extent, depend on the large-scale reorganization of the nuclear landscape. We have used electron spectroscopic imaging (ESI) to visualize chromatin organization at the mesoscale level of resolution in both pluripotent and differentiated cell types. Pluripotent cells are characterized by a highly dispersed mesh of 10-nm chromatin fibers that fill the nuclear volume. In contrast, differentiated cells display a propensity to form compact chromatin domains that lead to large regions of the nucleus devoid of DNA. Surprisingly, ESI combined with tomography methods reveals that the compact chromatin domains consist of 10-nm rather than 30-nm chromatin fibers. We propose that the transition between compact silent chromatin and open transcriptionally poised or active chromatin is based on the modulation of the packing density of 10-nm fibers rather than a transition between 10- and 30-nm fiber types.

Redox sensing: orthogonal control in cell cycle and apoptosis signalling.

Living systems have three major types of cell signalling systems that are dependent upon high-energy chemicals, redox environment and transmembranal ion-gating mechanisms. Development of integrated systems biology descriptions of cell signalling require conceptual models incorporating all three. Recent advances in redox biology show that thiol-disulphide redox systems are regulated under dynamic, nonequilibrium conditions, progressively oxidized with the life cycle of cells and distinct in terms of redox potentials amongst subcellular compartments. This article uses these observations as a basis to distinguish 'redox-sensing' mechanisms, which are more global biologic redox control mechanisms, from 'redox signalling', which involves conveyance of discrete activating or inactivating signals. Both redox sensing and redox signalling use sulphur switches, especially cysteine (Cys) residues in proteins which are sensitive to reversible oxidation, nitrosylation, glutathionylation, acylation, sulfhydration or metal binding. Unlike specific signalling mechanisms, the redox-sensing mechanisms provide means to globally affect the rates and activities of the high-energy, ion-gating and redox-signalling systems by controlling sensitivity, distribution, macromolecular interactions and mobility of signalling proteins. Effects mediated through Cys residues not directly involved in signalling means redox-sensing control can be orthogonal to the signalling mechanisms. This provides a capability to integrate signals according to cell cycle and physiologic state without fundamentally altering the signalling mechanisms. Recent findings that thiol-disulphide pools in humans are oxidized with age, environmental exposures and disease risk suggest that redox-sensing thiols could provide a central mechanistic link in disease development and progression.

Redox compartmentalization and cellular stress.

Mammalian cells are highly organized to optimize function. For instance, oxidative energy-producing processes in mitochondria are sequestered away from plasma membrane redox signalling complexes and also from nuclear DNA, which is subject to oxidant-induced mutation. Proteins are unique among macromolecules in having reversible oxidizable elements, 'sulphur switches', which support dynamic regulation of structure and function. Accumulating evidence shows that redox signalling and control systems are maintained under kinetically limited steady states, which are highly displaced from redox equilibrium and distinct among organelles. Mitochondria are most reducing and susceptible to oxidation under stressed conditions, while nuclei are also reducing but relatively resistant to oxidation. Within compartments, the glutathione and thioredoxin systems serve parallel and non-redundant functions to maintain the dynamic redox balance of subsets of protein cysteines, which function in redox signalling and control. This organization allows cells to be poised to respond to cell stress but also creates sites of vulnerability. Importantly, disruption of redox organization is a common basis for disease. Research tools are becoming available to elucidate details of subcellular redox organization, and this development highlights an opportunity for a new generation of targeted antioxidants to enhance and restore redox signalling and control in disease prevention.

Identification of a molecular recognition feature in the E1A oncoprotein that binds the SUMO conjugase UBC9 and likely interferes with polySUMOylation.

Hub proteins have central roles in regulating cellular processes. By targeting a single cellular hub, a viral oncogene may gain control over an entire module in the cellular interaction network that is potentially comprised of hundreds of proteins. The adenovirus E1A oncoprotein is a viral hub that interacts with many cellular hub proteins by short linear motifs/molecular recognition features (MoRFs). These interactions transform the architecture of the cellular protein interaction network and virtually reprogram the cell. To identify additional MoRFs within E1A, we screened portions of E1A for their ability to activate yeast pseudohyphal growth or differentiation. This identified a novel functional region within E1A conserved region 2 comprised of the sequence EVIDLT. This MoRF is necessary and sufficient to bind the N-terminal region of the SUMO conjugase UBC9, which also interacts with SUMO noncovalently and is involved in polySUMOylation. Our results suggest that E1A interferes with polySUMOylation, but not with monoSUMOylation. These data provide the first insight into the consequences of the interaction of E1A with UBC9, which was initially described in 1996. We further demonstrate that polySUMOylation regulates pseudohyphal growth and promyelocytic leukemia body reorganization by E1A. In conclusion, the interaction of the E1A oncogene with UBC9 mimics the normal binding between SUMO and UBC9 and represents a novel mechanism to modulate polySUMOylation.

Cerebral arterial oxygen saturation measurements using a fiber-optic pulse oximeter.

BACKGROUND: A pilot investigation was undertaken to assess the performance of a novel fiber-optic cerebral pulse oximetry system. A fiber-optic probe designed to pass through the lumen of a cranial bolt of the type used to make intracranial pressure measurements was used to obtain optical reflectance signals directly from brain tissue. METHODS: Short-duration measurements were made in six patients undergoing neurosurgery. These were followed by a longer duration measurement in a patient recovering from an intracerebral hematoma. Estimations of cerebral arterial oxygen saturation derived from a frequency domain-based algorithm are compared with simultaneous pulse oximetry (SpO2) and hemoximeter (SaO2) blood samples. RESULTS: The short-duration measurements showed that reliable photoplethysmographic signals could be obtained from the brain tissue. In the long-duration study, the mean (±SD) difference between cerebral oxygen saturation (ScaO2) and finger SpO2 (in saturation units) was -7.47(±3.4)%. The mean (±SD) difference between ScaO2 and blood SaO2 was -7.37(±2.8)%. CONCLUSIONS: This pilot study demonstrated that arterial oxygen saturation may be estimated from brain tissue via a fiber-optic pulse oximeter used in conjunction with a cranial bolt. Further studies are needed to confirm the clinical utility of the technique.

Structure and function of the perinucleolar compartment in cancer cells.

The perinucleolar compartment (PNC) is a subnuclear body that forms in cancer cells. In vivo analyses using human tumor tissues demonstrate a close correlation between PNC prevalence and disease progress in colorectal carcinoma, and a high PNC prevalence is associated with poor patient outcome. These findings are consistent with previous observations in breast cancer and cancer cell lines in vitro. The PNC is composed of thick strands that form a filamental meshwork often extending into the nucleolus. Although it appears to be electron dense as observed by transmission electron microscopy (TEM), the actual density of the structure imaged by electron spectroscopy is much lower, similar to that of the interchromatin space, and is lined with ribonucleoproteins (RNPs). In situ detections show that the PNC is highly enriched with a subset of small RNAs of polymerase III (Pol III) origins and RNA-binding proteins primarily implicated in pre-mRNA processing. A novel gel-shifting approach demonstrates that the addition of PNC-associated RNAs into HeLa cell lysates increases the mobility of polypyrimidine tract-binding (PTB) protein in a native gel electrophoresis, suggesting an interaction between these RNAs and PTB proteins. On the basis of these and other findings, we propose a working model in which novel RNPs have a key role in regulating gene expression at the PNC in cancer cells.

The Morscher Press Fit acetabular component: a nine- to 13-year review.

We reviewed the results at nine to 13 years of 125 total hip replacements in 113 patients using the monoblock uncemented Morscher press-fit acetabular component. The mean age at the time of operation was 56.9 years (36 to 74). The mean clinical follow-up was 11 years (9.7 to 13.5) and the mean radiological follow-up was 9.4 years (7.7 to 13.1). Three hips were revised, one immediately for instability, one for excessive wear and one for deep infection. No revisions were required for aseptic loosening. A total of eight hips (7.0%) had osteolytic lesions greater than 1 cm, in four around the acetabular component (3.5%). One required bone grafting behind a well-fixed implant. The mean wear rate was 0.11 mm/year (0.06 to 0.78) and was significantly higher in components with a steeper abduction angle. Kaplan-Meier survival curves at 13 years showed survival of 96.8% (95% confidence interval 90.2 to 99.0) for revision for any cause and of 95.7% (95% confidence interval 88.6 to 98.4) for any acetabular re-operation.

Preliminary evaluation of a new fibre-optic cerebral oximetry system.

A new system for measuring the oxygen saturation of blood within tissue has been developed, for a variety of patient monitoring applications. A particular unmet need is in the central nervous system, and this project aims to devise a means for measuring blood oxygen saturation in the brain tissue of patients recovering from neurosurgery or head injury. Coupling light sources and a photodetector to optical fibres results in a probe small enough to pass through a cranial bolt of the type already in use for intra-cranial pressure monitoring. The development and evaluation of a two-wavelength fibre-optic reflectance photoplethysmography (PPG) system are described. It was found that good quality red and near-infrared PPG signals could be obtained from the finger using a fibre-optic probe. Experiments were conducted to find the inter-fibre spacings that yield signals most suitable for calculating oxygen saturation. Reliable signals could be obtained for inter-fibre spacings between 2 mm and 5 mm, the latter being the size of the maximum aperture in the cranial bolt. A preliminary measurement from human brain tissue is also presented.

Tetrahydrobiopterin: a novel antihypertensive therapy.

Tetrahydrobiopterin (BH(4)) is a cofactor for the nitric oxide (NO) synthase enzymes, such that its insufficiency results in uncoupling of the enzyme, leading to release of superoxide rather than NO in disease states, including hypertension. We hypothesized that oral BH(4) will reduce arterial blood pressure (BP) and improve endothelial function in hypertensive subjects. Oral BH(4) was given to subjects with poorly controlled hypertension (BP >135/85 mm Hg) and weekly measurements of BP and endothelial function made. In Study 1, 5 or 10 mg kg(-1) day(-1) of BH(4) (n=8) was administered orally for 8 weeks, and in Study 2, 200 and 400 mg of BH(4) (n=16) was given in divided doses for 4 weeks. Study 1: significant reductions in systolic (P=0.005) and mean BP (P=0.01) were observed with both doses of BH(4). Systolic BP was 15+/-15 mm Hg (P=0.04) lower after 5 weeks and persisted for the 8-week study period. Study 2: subjects given 400 mg BH(4) had decreased systolic (P=0.03) and mean BP (P=0.04), with a peak decline of 16+/-19 mm Hg (P=0.04) at 3 weeks. BP returned to baseline 4 weeks after discontinuation. Significant improvement in endothelial function was observed in Study 1 subjects and those receiving 400 mg BH(4). There was no significant change in subjects given the 200 mg dose. This pilot investigation indicates that oral BH(4) at a daily dose of 400 mg or higher has a significant and sustained antihypertensive effect in subjects with poorly controlled hypertension, an effect that is associated with improved endothelial NO bioavailability.

Filtering techniques for the removal of ventilator artefact in oesophageal pulse oximetry.

The oesophagus has been shown to be a reliable site for monitoring blood oxygen saturation (SpO(2)). However, the photoplethysmographic (PPG) signals from the lower oesophagus are frequently contaminated by a ventilator artefact making the estimation of SpO(2) impossible. A 776th order finite impulse response (FIR) filter and a 695th order interpolated finite impulse response (IFIR) filter were implemented to suppress the artefact. Both filters attenuated the ventilator artefact satisfactorily without distorting the morphology of the PPG when processing recorded data from ten cardiopulmonary bypass patients. The IFIR filter was the better since it conformed more closely to the desired filter specifications and allowed real-time processing. The average improvements in signal-to-noise ratio (SNR) achieved by the FIR and IFIR filters for the fundamental component of the red PPG signals with respect to the fundamental component of the artefact were 57.96 and 60.60 dB, respectively. The corresponding average improvements achieved by the FIR and IFIR filters for the infrared PPG signals were 54.83 and 60.96 dB, respectively. Both filters were also compared with their equivalent tenth order Butterworth filters. The average SNR improvements for the FIR and IFIR filters were significantly higher than those for the Butterworth filters.

Risk factors for recurrence of maltreatment: a systematic review.

BACKGROUND: Children who have been maltreated are at increased risk of further maltreatment. Competent identification of those at highest risk of further maltreatment is an important part of safe and effective practice, but is a complex and demanding task. AIM: To systematically review the research base predicting those children at highest risk of recurrent maltreatment. METHODS: Systematic review of cohort studies investigating factors associated with substantiated maltreatment recurrence in children. RESULTS: Sixteen studies met the inclusion criteria. The studies were heterogeneous. A variety of forms of maltreatment were considered. Four factors were most consistently identified as predicting future maltreatment: number of previous episodes of maltreatment; neglect (as opposed to other forms of maltreatment); parental conflict; and parental mental health problems. Children maltreated previously were approximately six times more likely to experience recurrent maltreatment than children who had not previously been maltreated. The risk of recurrence was highest in the period soon after the index episode of maltreatment (within 30 days), and diminished thereafter. CONCLUSIONS: There are factors clearly associated with an increased risk of recurrent maltreatment, and these should be considered in professional assessments of children who have been maltreated. A comprehensive approach to risk assessment, including but not solely based on these factors, is likely to lead to interventions which offer greater protection to children.

The taurine transporter: mechanisms of regulation.

Taurine transport undergoes an adaptive response to changes in taurine availability. Unlike most amino acids, taurine is not metabolized or incorporated into protein but remains free in the intracellular water. Most amino acids are reabsorbed at rates of 98-99%, but reabsorption of taurine may range from 40% to 99.5%. Factors that influence taurine accumulation include ionic environment, electrochemical charge, and post-translational and transcriptional factors. Among these are protein kinase C (PKC) activation and transactivation or repression by proto-oncogenes such as WT1, c-Jun, c-Myb and p53. Renal adaptive regulation of the taurine transporter (TauT) was studied in vivo and in vitro. Site-directed mutagenesis and the oocyte expression system were used to study post-translational regulation of the TauT by PKC. Reporter genes and Northern and Western blots were used to study transcriptional regulation of the taurine transporter gene (TauT). We demonstrated that (i) the body pool of taurine is controlled through renal adaptive regulation of TauT in response to taurine availability; (ii) ionic environment, electrochemical charge, pH, and developmental ontogeny influence renal taurine accumulation; (iii) the fourth segment of TauT is involved in the gating of taurine across the cell membrane, which is controlled by PKC phosphorylation of serine 322 at the post-translational level; (iv) expression of TauT is repressed by the p53 tumour suppressor gene and is transactivated by proto-oncogenes such as WT1, c-Jun, and c-Myb; and (v) over-expression of TauT protects renal cells from cisplatin-induced nephrotoxicity.

Basal thumb metacarpal osteotomy for trapeziometacarpal osteoarthritis.

PURPOSE: To review the subjective and functional results of basal thumb metacarpal osteotomy for the treatment of trapeziometacarpal osteoarthritis. METHODS: Between July 1993 and November 1998, 35 thumb osteotomies without internal fixation were performed on 33 patients in the Christchurch Hospital, New Zealand. Records of 28 thumbs (13 right and 15 left) of 26 patients (17 women and 9 men) were available for review. Patients were reviewed using strength testing and the Michigan Hand Outcomes Questionnaire. RESULTS: The mean age of the 26 patients was 54 years (range, 30-69 years). Of the 28 thumbs, 22 (21 patients) had good or excellent results, 2 fair, one poor. The remaining 3 thumbs (3 patients) required further revision and were classified as failures. The mean follow-up period of the 25 thumbs (24 patients) not requiring revision was 34 months (range, 12-73 months). Good thumb motion was present in all hands with no trapeziometacarpal instability seen. Compared with the normative data, the strengths of key pinch, pulp pinch, and tripod pinch of our patients were significantly lower (22-32% lower), but not the grip strength. Michigan Hand Outcomes Questionnaire scores increased 28 (range, 1-56) points after surgery, with significant improvement especially in pain (+44 points), activities of daily living (one-handed tasks, +41 points), and satisfaction (+35 points). CONCLUSION: Basal thumb metacarpal osteotomy is a straightforward, conservative procedure that should be considered for grades II and III trapeziometacarpal osteoarthritis.

Long-term follow-up of a recurrent multifocal desmoid tumour treated with tamoxifen: a case report.

We report a long-term follow-up of a female patient with a multifocal extremity desmoid tumour. She had 3 local recurrences after excision and developed a second unresectable pelvic tumour that has remained unchanged in size for 14 years since starting tamoxifen treatment.

Evaluation of oesophageal reflectance pulse oximetry in major burns patients.

Pulse oximetry is being used in everyday clinical practice in anaesthesia utilising a peripheral probe. However, it may be unreliable in certain clinical situations such as hypothermia, hypovolemia, vasoconstriction and decreased cardiac output. Similar situations occur in burns patients and, more importantly, burns to extremities which limit the sites available for measurement of peripheral oxygen saturation (SpO(2)). To overcome these limitations, the oesophagus has been investigated as an alternative measurement site, as perfusion may be preferentially preserved centrally. A miniaturised reflectance oesophageal saturation (SpO(2)) probe has been constructed utilising infrared and red photodiodes and a photodetector. Our study was aimed at evaluating the reliability of oesophageal pulse oximetry in major burns patients. Seven adult patients (five males, two females) were studied. They were sedated and ventilated as part of their routine care. Measurable photoplethysmographic (PPG) traces and SpO(2) values were obtained in the oesophagus of all patients at a mean depth of 15.6+/-1.8 cm (measured from the lips). It was found that the oesophageal pulse oximeter results were in good agreement with oxygen saturation measurements obtained by a CO-oximeter. The mean (+/-S.D.) of the differences between the oesophageal oxygen saturation results and those from CO-oximetry was 0.50+/-0.69%. A Bland and Altman analysis showed that the bias and the limits of agreement between the oesophageal and commercial toe pulse oximeters were 0.4% and -3.6% to 4.6%, respectively. This study suggests that the oesophagus can be used as an alternative site for monitoring arterial blood oxygen saturation by pulse oximetry in burned patients.