Isolation and characterization of heparan sulfate from crude porcine intestinal mucosal peptidoglycan heparin.

A method for the preparation of heparan sulfate from peptidoglycan heparin is described. The objective of this research was to provide a basis for the development and validation of an industrial process to support the preclinical development of heparan sulfate and/or heparan sulfate derivatives. In the preparation of heparan sulfate, heparin was recovered by alcohol fractionation and dermatan sulfate was isolated by selective precipitation. The remaining crude heparan sulfate was fractionated by anion-exchange chromatography into five subfractions. The biological activities of these subfractions were examined by anticoagulant and amidolytic assays. Molecular weight and molecular size were determined using capillary viscometry and polyacrylamide gel electrophoresis. Charge density and degree of sulfation were determined by cellulose acetate electrophoresis and elemental analysis. Oligosaccharide and disaccharide analysis relied on enzymatic depolymerization using heparin lyases followed by polyacrylamide gel and capillary electrophoresis. 1H NMR analysis provided detailed structural information on each subfraction. Crude heparin sulfate and its subfractions showed significant differences in physical, structural and biological properties.

The prehospital treatment of pediatric trauma.

Prehospital pediatric trauma care is an important part of the EMS system. Review of 458 pediatric ALS trauma responses over two years treated in an urban, tiered ALS system revealed a male predominance. Violence (gunshot, stab, or assault) accounted for 46% of injuries, followed by vehicular accidents (occupant or pedestrian), with 35%. Important ALS resuscitation interventions were commonly performed en route, with a high degree of success (IVs = 93%, intubation = 79%), and did not greatly prolong field times (9 min BLS vs 11.7 min ALS). ALS procedure success rates and field times reported here are lower than previously described. Benchmark standards for the prehospital care of pediatric trauma are proposed.

Post-traumatic stress disorder in an urban trauma population.

This descriptive study examined the incidence of post-traumatic stress disorder (PTSD) in individuals who experienced physical traumas within the past 6 months and the relationship between severity of physical injury and PTSD. The sample consisted of 92 subjects who were treated at a Level 1 trauma center in an urban hospital. Scores on the Impact of Event Scale indicated that over half of the sample had medium or high levels of PTSD. However, the severity of physical injury from trauma was not related to PTSD development.

An evaluation of three blood pressure methods in a stabilized acute trauma population.

Blood pressure values were evaluated in 30 acute adult trauma patients using two indirect methods, the bell and diaphragm components of the stethoscope, and one direct arterial method. K1, K4, and K5 measurements were taken 4 to 72 hours (M = 30) after admission when vital signs had stabilized. Data were collected using a random-zero (RZ) sphygmomanometer for indirect blood pressure and radial intraarterial cannula for direct blood pressure. The sequence of measurement was randomized and variables shown to alter blood pressure values were controlled. There was no overall significant difference in K1, K4, and K5 blood pressures among the three methods. These results suggest that when clinicians monitor variables such as the frequency response of arterial lines and peripheral vascular resistance, indirect blood pressure methods are reliable for use with stabilized trauma patients.

A comparison of intravenous access sites for bolus injections during shock and resuscitation after emergency room thoracotomy with and without aortic crossclamping.

Thoracotomy with aortic crossclamping is used to resuscitate trauma victims. Pharmacologic boluses are often given via intravenous lines of central, brachial, or femoral origin. This study was undertaken to determine the efficacy of intravenous access site on delivery of a bolus injection to the heart with thoracotomy and aortic crossclamping during shock and resuscitation. Six dogs were anesthetized, intubated, and underwent brachial and femoral venotomy and Swan-Ganz insertion (central). Baseline measurements of central, brachial, and femoral transit times for 10 cc cold saline were obtained via Swan-Ganz thermistor. Each animal then underwent thoracotomy, aortic crossclamping alone, hemorrhage to blood pressure (BP) 50 mm Hg for 30 minutes with aorta unclamped and then aorta crossclamped and resuscitation with lactated Ringer's Solution with aorta unclamped and then crossclamped. Femoral-Brachial Index (FBI) was determined by dividing femoral transit time by brachial transit time at each observation. The data suggest that femoral access significantly prolongs bolus transit time when compared with central or brachial access during aortic crossclamping in the euvolemic, shock, or aggressively resuscitated model. Brachial access is therefore the preferred route for bolus injection delivery in the emergency room thoracotomy with or without aortic crossclamping because it provides expedient bolus delivery equal to central access and superior to femoral access.

Transport of L-glucose by Rhodotorula glutinis.

The kinetics of L-glucose transport by Rhodotorula glutinis were studied over a 720-fold range of sugar concentrations. Analysis of the saturation isotherm revealed the presence of a one-carrier system for L-glucose in the plasma membrane of Rhodotorula glutinis. This carrier exhibited a km of 3.7 +/- 0.3 mM. D-Ribose was found to be a competitive inhibitor with a Ki of 19 +/- 1 mM. The results suggest that L-glucose is transported by the high-Km, D-ribose carrier. L-Glucose was transported against a concentration gradient and the transport was inhibited by the proton conductor 2,4-dinitrophenol.

Tube thoracostomy and trauma--antibiotics or not?

Controversy persists regarding the use of antibiotics in association with t tube thoracostomy for trauma patients. We conducted a prospective randomized study of patients requiring tube thoracostomy for pneumo- and/or hemothorax complicating blunt or penetrating thoracic trauma in an attempt to assess the efficacy of prophylactic antibiotic therapy. Fifty-eight patients were included in the study. The control group (Group I) included 28 patients who received no antibiotic therapy: the experimental group (Group II) included 30 patients who received cefoxitin (1.0 gm IV q 6 h) commencing before tube thoracostomy and terminating 12 hours after its removal. The incidence of infectious complications (pneumonia and/or empyema) was recorded. Among the patients not receiving antibiotics, eight of 28 (29%) developed infectious chest complications. Of the patients receiving antibiotics, there was one infectious complication (3%). This difference is statistically significant (p = 0.0227). Cultures demonstrated significant conversion from negative to positive both within each group and between groups. The organism most commonly recovered was S. aureus. Our findings strongly suggest that patients requiring tube thoracostomy for trauma, whether blunt or penetrating, should receive the benefit of systemic prophylactic antibiotic therapy.

The demand for adult outpatient services in the Bicol region of the Philippines.

The absence of demand analysis for primary health care services has hampered efforts to finance these services and to make them permanent parts of Third World medical systems. This paper introduces a demand model for adult outpatient services, describes the types of data required for estimating it, and presents the results of a preliminary estimation using data from a poor rural region of the Philippines. The results indicate that prices and distance are not nearly as important as determinants of demand in this sample as has usually been assumed by planners. There appears to be considerable room for full or partial financing of outpatient services from user fees.

A simplified technique for gastric and duodenal decompression for duodenal injuries.

A simplified effective method of the decompression for use after repair of duodenal injuries is described. A Moss esophagogastric decompression tube is inserted as a gastrostomy to provide decompression of stomach and duodenum through a single tube. In addition, the complications associated with long-term nasogastric tube drainage are avoided.

Traditional and modern health professionals and breast-feeding in the Philippines.

We analyze a survey designed to assess the knowledge and attitudes of 1,298 Filipino health professionals towards breast-feeding. Some of the issues examined are the benefits of breast-feeding, the psychophysiology of lactation, contraindications to breast-feeding, and the effects of drugs and contraceptives on lactation. We found a fairly low level of knowledge about breast-feeding, particularly of health contraindications.

Infant formula promotion and infant-feeding practices, Bicol region, Philippines.

A 1978 household survey and a 1981 survey of stores and health facilities document the availability of breast milk substitutes, promotion of infant food and formula through the medical sector, and the effects of such promotion on the infant-feeding practices of mothers in the Bicol region of the Philippines. Breast milk substitutes were universally available in the region. Infant food and formula companies were active in the modern medical sector with promotional efforts that included distributing free formula samples, providing pamphlets and posters, and organizing professional meetings. Industry activities had mixed effects on health professionals' knowledge of breast-feeding and attitudes towards the practice. Activities of multinationals tended not to affect mothers' breast-feeding behavior directly but increased the probability of their introducing breast milk substitutes within the first six months, and thus in practicing a program of mixed feeding.

Kinetics of D-glucose and 2-deoxy-D-glucose transport by Rhodotorula glutinis.

The yeast Rhodotorula glutinis (Rhodosporidium toruloides) is capable of accumulative transport of a wide variety of monosaccharides. Initial velocity studies of the uptake of 2-deoxy-D-glucose were consistent with the presence of at least two carriers for this sugar in the Rhodotorula plasma membrane. Non-linear regression analysis of the data returned maximum velocities of 0.8 +/- 0.2 and 2.0 +/- 0.2 nmol/min per mg (wet weight) and Km values of 18 +/- 4 and 120 +/- 20 microM, respectively, for the two carriers. Kinetic studies of D-glucose transport also revealed two carriers with maximum velocities of 1.1 +/- 0.4 and 2.4 +/- 0.4 nmol/min per mg (wet weight) and Km values of 12 +/- 3 and 55 +/- 12 microM. As expected, 2-deoxy-D-glucose was a competitive inhibitor of D-glucose transport. Ki values for the inhibition were 16 +/- 8 and 110 +/- 40 microM. These Ki values were in good agreement with the Km values for 2-deoxy-D-glucose transport. D-Xylose, the 5-deoxymethyl analog of D-glucose, appears to utilize the D-glucose/2-deoxy-D-glucose carriers. This pentose was observed to be a competitive inhibitor of D-glucose (Ki values = 0.14 +/- 0.06 and 5.6 +/- 1.6 mM) and 2-deoxy-D-glucose (Ki values = 0.15 +/- 0.07 and 4.6 +/- 1.2 mM) transport.

Transport and metabolism of 2-deoxy-D-glucose by Rhodotorula glutinis.

2-Deoxy-D-glucose transport by Rhodotorula glutinis is an active process. The intracellular concentration of free deoxyglucose after 15 min incubation of Rhodotorula cells with this sugar was 230 times the extracellular concentration. Although cell extracts at this time contained more 2-deoxy-D-glucose 6-phosphate than deoxyglucose, pulse-labelling experiments demonstrated that deoxyglucose is transported as the free sugar and subsequently phosphorylated. After transport, Rhodotorula cells metabolize deoxyglucose. The major metabolites during 30-90 min incubations were determined to be 2-deoxy-D-glucose 6-phosphate, 2-deoxy-D-glucitol, 2-deoxy-D-gluconate and 2,2'-dideoxy-alpha, alpha'-trehalose. Rhodotorula glutinis also degrades deoxyglucose to CO2. The concentrations of intermediates in this pathway were too low to detect and resolve in extracts of control cells. In 2,4-dinitrophenol-poisoned cells, however, it appears that deoxyglucose degradation is restricted largely to loss of C-1 as CO2 and it was possible to identify 1-deoxy-D-ribulose 5-phosphate as an intermediate presumably arising from metabolism of deoxyglucose by the oxidative portion of the hexose monophosphate pathway.

Carrier-mediated transport of D-ribose by Rhodotorula glutinis.

The kinetics of D-ribose transport by Rhodotorula glutinis were investigated over a 1 000-fold range of sugar concentrations. Analysis of the saturation isotherm revealed the presence of two carrier systems for D-ribose in the Rhodotorula plasma membrane. These two carriers exhibited Km values of 1.3 and 30 mM. At saturating concentrations of D-ribose, the low Km carrier contributes less than 10% to the total rate of transport. Although D-ribose is metabolized rapidly by Rhodotorula, intracellular free sugar concentrations exceed those in the medium. In addition, the transport of this pentose is inhibited by the proton-conductors, 2,4-dinitrophenol and carbonylcyanide m-chlorophenylhydrazone. These data suggest that Rhodotorula cells are capable of an energy-dependent, concentrative transport of D-ribose.

Phosphofructokinase from Escherichia coli: further evidence for identical subunits.

A procedure for the purification of Escherichia coli phosphofructokinase by affinity chromatography is described. The results of amino acid analyses of the purified protein and tryptic peptide mapping suggest that the tetrameric phosphofructokinase is composed of chemically identical subunits. In addition, the reaction product, ADP, was observed to bind to 4.1 +/- 0.1 equal and independent sites on the enzyme.

A kinetic analysis of D-xylose transport in Rhodotorula glutinis.

The kinetics of D-xylose transport were studied in Rhodotorula glutinis. Analysis of the saturation isotherm revealed the presence of at least two carriers for D-xylose in the Rhodotorula plasma membrane. These two carriers exhibited Km values differing by more than an order of magnitude. The low-Km carrier was repressed in rapidly growing cells and derepressed by starvation of the cells. Several hexoses were observed to inhibit D-xylose transport. In the studies reported here, the inhibitions produced by D-galactose and 2-deoxy-D-glucose were examined in some detail in order to define the interactions of these sugars with the D-xylose carriers. 2-Deoxy-D-glucose competitively inhibited both of the D-xylose carriers. In contrast, only the low-Km carrier was competitively inhibited by D-galactose.

Rapid determination of the amount of cetylpyridinium chloride bound by bacteria.

A modification of the Colowick and Womack procedure for measuring ligand binding by macromolecules is described for drug binding by bacteria. This technique is based on the determination of drug concentration in the dialysate from a bacteria-drug mixture at equilibrium. The dialysis cell of the original procedure was replaced by a Minibeaker (Bio-Rad), which has a larger membrane surface area, and the dialysate was continuously monitored with a spectrophotometer equipped with a flow cell. With this system, only 3 min was required to determine the amount of cetylpyridinium chloride bound by Escherichia coli K-12 strain P678. Possible applications of the technique are discussed.