Pilot Study on Music in the Waiting Room of Outpatient Pain Clinics.

BACKGROUND: This study explores live and recorded music listening in the outpatient pain clinic. There is evidence demonstrating the effectiveness of live and recorded music in a hospital setting but a comparison study of this kind has yet to be conducted. METHODS: A multimethod survey study design was used. A questionnaire utilizing rating scales was self-administered across two outpatient pain clinic waiting rooms. Patients were included through convenience sampling. In one clinic, a playlist of recorded music curated by two of the authors was provided. In the second clinic, a music therapy student played live music using guitar, flute, and voice. The questionnaire gathered data on music's impact on pain and emotional states, as well as attitudes toward music in the waiting room. Quantitative data was analyzed using descriptive statistics and qualitative data, gathered in the questionnaires open ended question, was analyzed using thematic analysis. RESULTS: The questionnaire was completed by 200 adult patients. Patients reported lowered levels of anxiety, stress, and pain in both clinics, as well as a shorter waiting time and more caring experience. Patients in the live music clinic reported that music lowered levels of stress, nervousness, agitation, and pain more than in the recorded music clinic. CONCLUSIONS: Participants in this study identified that music is a useful tool in the pain clinic waiting room. This study contributes to evidence that music should be considered as a complimentary treatment for people living with pain and in the wider hospital setting. Additional research is warranted with a control group, pre- and posttesting, and studies of music in hospitals in a range of cultural contexts.

Influence of feeding a fish oil-containing diet to mature, overweight dogs: Effects on lipid metabolites, postprandial glycaemia and body weight.

The objective of this study was to determine the effect of feeding a fish oil (FO)-containing diet on lipid and protein metabolism, postprandial glycaemia and body weight (BW) of mature, overweight dogs. Seven female dogs were randomly assigned to one of two isonitrogenous and isocaloric diets, control (CO) or FO (FO), in a crossover design. Experimental periods were 69 day, separated by a washout period of 30 day. At the beginning of the experiment, and at 30 and 60 day of feeding the experimental diets, the dogs were infused with D-glucose (2 g/kg BW) through an intravenous catheter. Blood samples were collected for 3 hr to perform a glucose tolerance test. Nitrogen balance measurements began at 06:30 on d 63 of each experimental period and ended at 06:30 on d 69. On d 66 of each period, a single dose (7.5 mg/kg) of 15 N-glycine was administered orally for determination of protein turnover. Incremental area under the curve and glucose concentration at peak did not differ between treatments or among sampling days within treatment. Glucose half-life tended to decrease (p < .10) in the FO treatment on day 30 when compared to baseline (day 0). ß-hydroxybutyrate, non-esterified fatty acid (NEFA) and triglycerides did not differ within or between treatments. Cholesterol decreased (p < .05) on the FO treatment on day 30, 60 and 69 when compared to day 0. High-density lipoprotein (HDL) decreased (p < .05) in the FO treatment on day 69 when compared to day 0. Body weight, food intake, faecal excretion, DM and N digestibilities, N balance and protein turnover were not different between diets. Overall, FO-containing diet decreases cholesterol in mature overweight dogs; however, further research is warranted to verify the effects of FO on glucose metabolism.

Prevention of multidrug resistance (MDR) in osteosarcoma by NSC23925.

BACKGROUND: The major limitation to the success of chemotherapy in osteosarcoma is the development of multidrug resistance (MDR). Preventing the emergence of MDR during chemotherapy treatment has been a high priority of clinical and investigational oncology, but it remains an elusive goal. The NSC23925 has recently been identified as a novel and potent MDR reversal agent. However, whether NSC23925 can prevent the development of MDR in cancer is unknown. Therefore, this study aims to evaluate the effects of NSC23925 on prevention of the development of MDR in osteosarcoma. METHODS: Human osteosarcoma cell lines U-2OS and Saos were exposed to increasing concentrations of paclitaxel alone or in combination with NSC23925 for 6 months. Cell sublines selected at different time points were evaluated for their drug sensitivity, drug transporter P-glycoprotein (Pgp) expression and activity. RESULTS: We observed that tumour cells selected with increasing concentrations of paclitaxel alone developed MDR with resistance to paclitaxel and other Pgp substrates, whereas cells cultured with paclitaxel-NSC23925 did not develop MDR and cells remained sensitive to chemotherapeutic agents. Paclitaxel-resistant cells showed high expression and activity of the Pgp, whereas paclitaxel-NSC23925-treated cells did not express Pgp. No changes in IC50 and Pgp expression and activity were observed in cells grown with the NSC23925 alone. CONCLUSIONS: Our findings suggest that NSC23925 may prevent the development of MDR by specifically preventing the overexpression of Pgp. Given the significant incidence of MDR in osteosarcoma and the lack of effective agents for prevention of MDR, NSC23925 and derivatives hold the potential to improve the outcome of cancer patients with poor prognosis due to drug resistance.

Informed consent for epidural analgesia in labour: a survey of Irish practice.

Currently, we do not have a national standard regarding epidural consent in Ireland. The aim of this survey was to assess practice in obstetric units in Ireland with regard to obtaining informed consent prior to epidural insertion, and whether the risks discussed with women are being documented. A postal survey of anaesthetists in Irish obstetric units was performed in January 2012 to assess practice regarding obtaining informed consent prior to epidural insertion, and documentation of the risks discussed. The response rate was 16/18 (88%). There was major variation both in which risks are discussed with women in labour and what risks are quoted. The most frequently quoted risks were headache--15/16 (93.8% of the respondents), partially/not working epidural--15/16 (93.8%), drop in blood pressure--14/16 (87.5%) and temporary backache/local tenderness--12/16 (75%). The more serious risks were not discussed as frequently: permanent nerve damage--8/16 (50%), paralysis--8/16 (50%), epidural abscess/haematoma--6/16 (37.5%), meningitis--3/16 (18.7%). The vast majority of respondents supported introduction of a national standardised information leaflet, detailing all the benefits and risks of epidural analgesia, to be shown to all women before consenting to epidural insertion.

The role of perceived patient injustice on pain related and functional outcomes in orthopaedic patients; a systematic review.

Introduction: Higher levels of perceived patient injustice has been associated with higher levels of pain and poor functional outcomes in patients with chronic musculoskeletal pain. We wanted to investigate if there was any evidence of this association in patients who underwent orthopaedic surgery. Materials and methods: A systematic search of the literature was performed independently by two researchers on the electronic databases of MEDLINE, EMBASE, Google Scholar, Web of Science, and the Cochrane Database of Systematic Reviews and data extracted in accordance with PRISMA guidelines for systematic reviews. Outcomes of interest were pain, pain intensity, pain interference, opiate use, disability, physical function, return to work, quality of life, depression, anxiety, post-traumatic stress, social functioning and quality of life. Results: Five studies were identified for qualitative analysis involving elective and trauma orthopaedic patients. All studies identified a positive correlation between higher levels of perceived injustice and depression, pain intensity and interference as well as reduced function and higher levels of disability. Conclusions: Higher levels of perceived patient injustice is associated with worse pain and functional outcomes in patients who undergo orthopaedic surgery in the traumatic and elective setting. Further research is warranted to further elucidate this association and identify potential therapeutic interventions.

Thromboembolic Events Associated with Thalidomide and Multimodality Therapy for Soft Tissue Sarcomas: Results of RTOG 0330.

Introduction. RTOG 0330 was developed to address the toxicity of RTOG 9514 and to add thalidomide (THAL) to MAID chemoradiation for intermediate/high grade soft tissue sarcomas (STSs) and to preoperative radiation (XRT) for low-grade STS. Methods. Primary/locally recurrent extremity/trunk STS: ≥8 cm, intermediate/high grade (cohort A): >5 cm, low grade (cohort B). Cohort A: 3 cycles of neoadjuvant MAID, 2 cycles of interdigitated THAL (200 mg/day)/concurrent 22 Gy XRT, resection, 12 months of adjuvant THAL. Cohort B: neoadjuvant THAL/concurrent 50 Gy XRT, resection, 6 months of adjuvant THAL. Planned accrual 44 patients. Results. 22 primary STS patients (cohort A/B 15/7). Cohort A/B: median age of 49/47 years; median tumor size 12.8/10 cm. 100% preoperative THAL/XRT and surgical resection. Three cycles of MAID were delivered in 93% cohort A. Positive margins: 27% cohort A/29% cohort B. Adjuvant THAL: 60% cohort A/57% cohort B. Grade 3/4 venous thromboembolic (VTE) events: 40% cohort A (1 catheter thrombus and 5 DVT or PE) versus 0% cohort B. RTOG 0330 closed early due to cohort A VTE risk and cohort B poor accrual. Conclusion. Neoadjuvant MAID with THAL/XRT was associated with increased VTE events not seen with THAL/XRT alone or in RTOG 9514 with neoadjuvant MAID/XRT.

Glucagon-like peptide-2 (GLP-2) increases small intestinal blood flow and mucosal growth in ruminating calves.

Glucagon-like peptide-2 (GLP-2) increases small intestinal mass and blood flow in nonruminants but its effect in ruminants is unknown. Eight Holstein calves with an ultrasonic flow probe around the superior mesenteric artery and catheters in the carotid artery and mesenteric vein were paired by age and randomly assigned to treatment of a control (0.5% of BSA in saline; n=4) or GLP-2 (50 µg/kg of body weight of bovine GLP-2 in BSA; n=4) given subcutaneously every 12h for 10 d. Blood flow was measured on d 0 (acute) and d 10 (chronic) and included 3 periods: baseline (saline infusion), treatment (infusion of BSA or 1,000 pmol of GLP-2/kg of body weight per h), and recovery (saline infusion). On d 11, calves were killed 2h after injection of 5-bromo-2'-deoxyuridine (BrdU). Gastrointestinal tissues were weighed and epithelial samples were obtained to determine villus height, crypt depth, and BrdU staining. Infusion of GLP-2 increased superior mesenteric artery blood flow to 175% of baseline on d 0 but to only 137% of baseline after chronic treatment. Compared with that of the control, GLP-2 increased small intestinal mass by 24% by increasing epithelial mass in the jejunum and ileum. Additionally, GLP-2 increased villus height, crypt depth, and BrdU-labeling in small intestinal segments. These results demonstrate that GLP-2 induces similar increases in small intestinal blood flow and growth in ruminants to those observed in nonruminants. Furthermore, GLP-2 increases small intestinal blood flow in ruminants but this response is attenuated after 10 d of GLP-2 administration. In cattle, GLP-2 may be an important hormone in the regulation of intestinal blood flow and epithelial growth.

5-Hydroxytryptophan strongly stimulates serotonin synthesis in Holstein steers.

Although serotonin has been extensively studied in many species, there is a lack of information in ruminants, and no research has been evaluated if its precursor, 5-hydroxytryptophan (5-HTP), administered into the abomasum may be used as a means to manipulate serotonin metabolism. Thus, the objective of this study was to evaluate if intra-abomasal infusion of 5-HTP increases circulating serotonin in the steer. Eight Holstein steers (471 ± 8.9 kg) were used in a replicated 4 × 4 Latin Square design experiment. The treatments were intra-abomasal infusion of 5-HTP at 0.5, 1, 2.5, and 5 mg/kg BW. Blood was collected at 0, 2, 4, 6, 8, and 24 h after infusion. The serum concentration of 5-HTP increased quadratically (P = 0.005) with a peak at 2 h after administration. The 5-HTP administration increased (P < 0.05) serum serotonin in comparison with baseline with no difference (P > 0.05) between the doses of 5-HTP. When 5-HTP was dosed at 2.5 mg/kg BW or higher, intake decreased, and there was an altered manure consistency. The serum 5-hydroxyindole acetic acid concentrations followed the same pattern as 5-HTP. Plasma glucose content was not affected (P > 0.05) by 5-HTP dosing. However, free fatty acids concentration in the plasma was lower (P > 0.05) compared with baseline for the infusion levels of 0.5 and 1 mg/kg BW. Intra-abomasal infusion of 5-HTP efficiently increases serum serotonin cattle.

Residual effects of abomasal 5-hydroxytryptophan administration on serotonin metabolism in cattle.

Studies of serotonin in animal husbandry has received growing interest. However, there is limited information about serotonin manipulation using 5-HTP administered postruminally and its residual effects in cattle. The objective of this study was to evaluate the effectiveness of 5-HTP infused into the abomasum for enhancing circulating serotonin in cattle. Four Holstein steers (487 ± 7.6 kg) fitted with ruminal cannulas were used in a 4 × 4 Latin Square design experiment. The treatments were intra-abomasal infusion of 5-HTP at 0, 0.25, 0.5, and 1 mg/kg BW. Blood was collected from the jugular vein of each steer at -60, -30, 0, 30, 60, 120, 240, and 480 min from 5-HTP infusion for basal and short term evaluation and, at 1, 2, 4, and 7 d after 5-HTP infusion for long term evaluation. Dry matter intake was not affected (P > 0.05) by intra-abomasal infusions. The half-life of 5-HTP was dose-independent (128 min). The serum 5-HTP, serotonin, and 5-hydroxyindoleacetic acid area under the curve increased (P < 0.05) linearly with an increased dose of 5-HTP. Serum 5-HTP reached peak concentration in approximately 30 min after dosing while serum and plasma serotonin peaked after 240 min postinfusion. Serotonin was greater than control for all 5-HTP doses 1 d and 2 d after infusion in serum and plasma, respectively. Intra-abomasal infusion of 5-HTP at doses up to 1 mg/ kg BW increases circulating serotonin for up 2 days.

Pattern of postruminal administration of l-tryptophan affects blood serotonin in cattle.

Serotonin (5-HT) has many important functions in both central and peripheral nervous systems. Although it has been demonstrated that manipulation of serotonin metabolism is possible in many species, there is limited information about l-tryptophan (TRP), a serotonin precursor, in cattle, and these provide conflicting results. Furthermore, there is no study evaluating how different patterns of intra-abomasal infusion of TRP impact circulating 5-HT. The objective of this study was to evaluate if intra-abomasal infusion patterns of TRP can affect circulating 5-HT and other metabolites from TRP metabolism in the plasma and serum and circulating glucose and insulin in cattle. Eight ruminally cannulated Holstein steers were used in a replicated 4 × 4 Latin square design. Each received intra-abomasal water infusion (control) or intra-abomasal TRP infusion (50 mg/kg BW) in 3 different patterns: a pulse infusion once a day (pulse once), pulse infusion twice a day (pulse twice), or continuous infusion (continuous). For continuous treatment, the TRP dose was diluted in tap water and infused by a peristaltic pump (300 mL/h). To equalize conditions, the other treatments had a water infusion (300 mL/h). The steers were fed every 2 h, and blood was collected from a jugular vein catheter every 4 h for 24 h after the initial infusion. Urine produced during the 24 h period was collected. Serum and plasma TRP, 5-HT and kynurenine, plasma glucose, and serum insulin concentrations were analyzed. Urine was analyzed for concentrations of 5-hydroxyindoleacetic acid. Both serum TRP and kynurenine were increased (P < 0.05) by all TRP infusion treatments, but concentrations in pulse dose treatments were greater than those in continuous infusion. Serum 5-HT increased (P < 0.05) with both pulse TRP infusion treatments; however, the continuous TRP infusion did not increase the serum 5-HT. Plasma 5-HT, glucose, and insulin had a tendency to increase with TRP pulse infusions. The urinary 5-hydroxyindoleacetic acid excretion was highest for pulse dose treatments. An acute supply of TRP in 1 or 2 daily doses increases serum 5-HT and increases circulating glucose and insulin in cattle. The TRP and kynurenine concentrations are similar in plasma and serum. However, the serum 5-HT concentration is more responsive to TRP administration than plasma.

Technical note: ruminal cannulation technique in young Holstein calves: effects of cannulation on feed intake, body weight gain, and ruminal development at six weeks of age.

Ruminal cannulation techniques are frequently used to study fermentation in the ruminant forestomach. Unsatisfactory results with the traditionally applied procedure for cannulation of young calves stimulated the development of a simpler and more robust procedure; this procedure was tested for effects on performance traits and gross anatomy of the gastrointestinal tract compared with a control group not undergoing surgery. Five calves were ruminally cannulated at approximately 10 d of age and 5 matching calves were used as controls. All calves were fed milk replacer and a diet based on clover grass silage and sodium hydroxide-treated wheat. Ruminal fluid was collected from cannulated calves once weekly for 3 consecutive weeks. All calves were euthanized at 43+/-3 d of age. No apparent adverse effects of cannulation were observed. Feed intake, BW gain, and gross anatomy of the gastrointestinal tract were not affected by cannulation. Minimum ruminal pH increased with sampling week, but average ruminal pH, total volatile fatty acids concentration, and volatile fatty acids proportions were not affected by sampling week. In conclusion, the implemented surgical technique was found to have no major effect on apparent animal health and performance traits, and the cannula proved useful for multiple samplings of ruminal contents in young calves.

Review: Nutritional regulation of intestinal starch and protein assimilation in ruminants.

Pregastric fermentation along with production practices that are dependent on high-energy diets means ruminants rely heavily on starch and protein assimilation for a substantial portion of their nutrient needs. While the majority of dietary starch may be fermented in the rumen, significant portions can flow to the small intestine. The initial phase of small intestinal digestion requires pancreatic α-amylase. Numerous nutritional factors have been shown to influence pancreatic α-amylase secretion with starch producing negative effects and casein, certain amino acids and dietary energy having positive effects. To date, manipulation of α-amylase secretion has not resulted in substantial changes in digestibility. The second phase of digestion involves the actions of the brush border enzymes sucrase-isomaltase and maltase-glucoamylase. Genetically, ruminants appear to possess these enzymes; however, the absence of measurable sucrase activity and limited adaptation with changes in diet suggests a reduced capacity for this phase of digestion. The final phase of carbohydrate assimilation is glucose transport. Ruminants possess Na+-dependent glucose transport that has been shown to be inducible. Because of the nature of pregastric fermentation, ruminants see a near constant flow of microbial protein to the small intestine. This results in a nutrient supply, which places a high priority on protein digestion and utilization. Comparatively, little research has been conducted describing protein assimilation. Enzymes and processes appear consistent with non-ruminants and are likely not limiting for efficient digestion of most feedstuffs. The mechanisms regulating the nutritional modulation of digestive function in the small intestine are complex and coordinated via the substrate, neural and hormonal effects in the small intestine, pancreas, peripheral tissues and the pituitary-hypothalamic axis. More research is needed in ruminants to help unravel the complexities by which small intestinal digestion is regulated with the aim of developing approaches to enhance and improve the efficiency of small intestinal digestion.

Ruminal and abomasal starch hydrolysate infusions selectively decrease the expression of cationic amino acid transporter mRNA by small intestinal epithelia of forage-fed beef steers.

Although cationic amino acids (CAA) are considered essential to maximize optimal growth of cattle, transporters responsible for CAA absorption by bovine small intestinal epithelia have not been described. This study was conducted to test 2 hypotheses: 1) the duodenal, jejunal, and ileal epithelia of beef cattle differentially express 7 mRNA associated with 4 mammalian amino acid (AA) transport activities: y(+) (CAT1), B(0,+) (ATB(0,+)), b(0,+) (b(0,+)AT and rBAT), and y(+)L (y(+)LAT1, y(+)LAT2, and 4F2hc), and 2) the expression of these mRNA is responsive to small intestinal luminal supply of AA substrates (derived from ruminal microbes) or glucose-derived energy (from starch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (body weight = 260 +/- 17 kg) fed an alfalfa cube-based diet at 1.33 x net energy for maintenance requirement were assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control); ruminal SH and abomasal water infusion; and ruminal water and abomasal SH infusion. The dosage of SH infusion amounted to 20% of metabolizable energy intake. After 14 or 16 d of infusion, steers were slaughtered, duodenal, jejunal, and ileal epithelia were harvested, and total RNA was extracted. The relative amounts of mRNA expressed by epithelia were quantified using real-time reverse transcription-PCR. All 7 mRNA species were expressed by the epithelium from each region, but their abundance differed among the regions. Specifically, duodenal expression of CAT1 and ATB(0,+) mRNA was greater than jejunal or ileal expression; ileal expression of b(0,+)AT, rBAT, and y(+)LAT1 mRNA was greater than jejunal or duodenal expression, whereas the expression of y(+)LAT2 and 4F2hc mRNA did not differ among the 3 epithelia. With regard to SH infusion effect, ruminal infusion down-regulated or tended to down-regulate the jejunal expression of CAT1, rBAT, y(+)LAT2, and 4F2hc mRNA. Abomasal infusion down-regulated the jejunal expression of y(+)LAT2 mRNA and tended to down-regulate the jejunal expression of 4F2hc mRNA. This study characterized the pattern of CAA transporter mRNA expressed by growing beef cattle fed an alfalfa-based diet. Moreover, this study demonstrated that increasing the luminal supply of microbe-derived AA (by ruminal supplementation of SH) results in a reduced capacity of apical and basolateral membrane to transport of CAA, whereas increasing luminal glucose supply (by abomasal supplementation of SH) reduces only the basolateral transport capacity, assuming that CAA transporter mRNA content represents functional capacity.

Influence of rumen escape starch on pancreatic exocrine secretion of goats.

The objective of the experiment was to evaluate the effect of rumen escape starch (RES), accomplished by altering dietary starch concentrations on pancreatic exocrine secretion of goats. Four goats (36.8 +/- 3.2 kg) with common bile duct re-entrant and duodenal catheters were used in a 4 x 4 Latin square. Goats were fed diets containing 20%, 30%, 40% and 50% starch. Periods consisted of 10 day adaptation followed by 3 day of sample collection. Juice was collected in 1-h fractions continuously for 72 h. Total juice secreted was recorded, and 3% sub samples were retained to form a composite sample. The remaining fluid was returned to the duodenum. Juice composite samples were analyzed for activities of alpha-amylase, trypsin, chymotrypsin and lipase. Secretion of pancreatic alpha-amylase was lower (p < 0.05) when comparing lambs fed 20% starch diet with 30%, 40% and 50% starch diets. Lipase secretion was greater (p < 0.05) in lambs fed 40% starch diet compared with the other diets. Total secretion of juice, trypsin and chymotrypin was not affected (p > 0.05) by dietary starch concentration. Rumen escape starch increased with increasing dietary starch concentration (p < 0.05). Regression analysis showed that increasing RES results in a quadratic increase (p < 0.05) in pancreatic alpha-amylase and lipase secretion, and the secretion of alpha-amylase and lipase is maximum when RES is 113 and 83 g/day respectively. These results suggest that optimal RES for pancreatic secretion of alpha-amylase and lipase is 80-110 g/day in adult goats.

Pulsed radiofrequency of the brachial plexus in the treatment of chemotherapy-induced peripheral neuropathy of the upper limb.

We describe the treatment of chemotherapy-induced peripheral neuropathy in the upper limb of a patient via ultrasound-guided pulsed radiofrequency of the brachial plexus. A 54-year-old female, who underwent chemotherapy and mastectomy for left-sided breast cancer, presented to the pain clinic describing continuous and severe shock-like pain in the posterolateral aspect of the left upper limb, above the elbow. A diagnosis of chemotherapy-induced peripheral neuropathy was made. Pain management with multi-modal analgesia was not fully effective and pulsed radiofrequency was offered as an alternative. Ultrasonography of the supraclavicular region was used to identify the brachial plexus, followed by pulsed radiofrequency using an insulated-tip needle that produced paraesthesia in the affected area. Onset of pain relief occurred one week post-procedure and lasted approximately 10 weeks. An estimated 80% decrease in pain intensity was reported by the patient. We propose that pulsed radiofrequency can be offered as a pain management alternative in certain presentations of chemotherapy-induced peripheral neuropathy.

Basal expression of nucleoside transporter mRNA differs among small intestinal epithelia of beef steers and is differentially altered by ruminal or abomasal infusion of starch hydrolysate.

In ruminants, microbial-derived nucleic acids are a major source of N and are absorbed as nucleosides by small intestinal epithelia. Although the biochemical activities of 2 nucleoside transport systems have been described for cattle, little is known regarding the regulation of their gene expression. This study was conducted to test 2 hypotheses: (1) the small intestinal epithelia of beef cattle differentially express mRNA for 3 concentrative (CNT1, 2, 3) and 2 equilibrative (ENT1, 2) nucleoside transporters (NT), and (2) expression of these NT is responsive to small intestine luminal supply of rumen-derived microbes (hence, nucleosides), energy (cornstarch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (260 +/- 17 kg of BW) were fed an alfalfa cube-based diet at 1.33x NE(m) requirement. Six steers in each of 3 periods were blocked by BW (heavy vs. light). Within each block, 3 steers were randomly assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control), ruminal SH infusion/abomasal water infusion, or ruminal water infusion/abomasal SH infusion. The dosage of SH infusion amounted to 20% of ME intake. After a 14-or 16-d infusion period, steers were slaughtered, and duodenal, jejunal, and ileal epithelia were harvested for total RNA extraction and the relative amounts of mRNA expressed were determined using real-time RT-PCR quantification methodologies. All 5 NT mRNA were found expressed by each epithelium, but their abundance differed among epithelia. Specifically, jejunal expression of all 5 NT mRNA was higher than that by the ileum, whereas jejunal expression of CNT1, CNT3, and ENT1 mRNA was higher, or tended to be higher, than duodenal expression. Duodenal expression of CNT2, CNT3, and ENT2 mRNA was higher than ileal expression. With regard to SH infusion treatments, ruminal infusion increased duodenal expression of CNT3 (67%), ENT1 (51%), and ENT2 (39%) mRNA and ileal expression of CNT3 (210%) and ENT2 (65%) mRNA. Abomasal infusion increased (54%) ileal expression of ENT2 mRNA and tended to increase (50%) jejunal ENT2 mRNA expression. This study has uniquely characterized the pattern of NT mRNA expression by growing beef cattle and found that the mRNA abundance for CNT3, ENT1, and ENT2 in small intestinal epithelia can be increased by increasing the luminal supply of nucleotides (CNT3, ENT1, ENT2) or glucose (ENT2).

The effect of midazolam on cerebral endothelial (P-selectin and ICAM-1) adhesion molecule expression during hypoxia-reperfusion injury in vitro.

BACKGROUND AND OBJECTIVE: Hypoxia-reperfusion injury is an important determinant of secondary brain injury. In the acute phase of cerebral reperfusion, pro-inflammatory events enhance expression of cerebral endothelial (intercellular adhesion molecule-1 and P-selectin) adhesion molecules, which play an important role in brain hypoxia-reperfusion injury. Midazolam is the most commonly used sedative in patients with brain injury. The objective of this investigation was to examine the effect of midazolam on the expression of cerebral endothelial intercellular adhesion molecule-1 and P-selectin during hypoxia-reperfusion injury in vitro. METHODS: The up-regulation of mouse cerebral endothelial cells intercellular adhesion molecule-1 and P-selectin was assessed following hypoxia-reoxygenation (hypoxia-reperfusion). Cells were pre-treated with three different concentrations of midazolam (0, 5 and 50 microg mL(-1)) prior to hypoxia. Flow cytometry was used to estimate adhesion molecule expression mean channel fluorescence. Data are presented as mean +/- SD. RESULTS: Mouse cerebral endothelial cell intercellular adhesion molecule-1 and P-selectin expression was greater after exposure to hypoxia-reoxygenation compared to normoxia (mean channel fluorescence) 241 +/- 12 vs. 140 +/- 7 and 120 +/- 14 vs. 46 +/- 7, respectively, P < 0.05. Intercellular adhesion molecule-1 and P-selectin expression was decreased by midazolam (5 microg mL(-1)) pre-incubation compared to control, mean channel fluorescence 184 +/- 10 vs. 241 +/- 12 and 51 +/- 7 vs. 120 +/- 14, respectively, P < 0.05. Midazolam at 50 microg mL(-1) had the same effect as 5 microg mL(-1). CONCLUSION: Pre-treatment with midazolam diminishes increased expression of cerebral endothelial intercellular adhesion molecule-1 and P-selectin expression following hypoxia-reoxygenation.