Collisional activation with random noise in ion trap mass spectrometry.

Random noise applied to the end caps of a quadrupole ion trap is shown to be an effective means for the collisional activation of trapped ions independent of mass/charge ratio and number of ions. This technique is compared and contrasted with conventional single-frequency collisional activation for the molecular ion of N,N-dimethylaniline, protonated cocaine, the molecular anion of 2,4,6-trinitrotoluene, and doubly pronated neuromedin U-8. Collisional activation with noise tends to produce more extensive fragmentation than the conventional approach due to the fact that product ions are also kinetically excited in the noise experiment. The efficiency of the noise experiment in producing detectable product ions relative to the conventional approach ranges from being equivalent to being a factor of 3 less efficient. Furthermore, discrimination against low mass/charge product ions is apparent in the data from multiply charged biomolecules. Nevertheless, collisional activation with random noise provides a very simple means for overcoming problems associated with the dependence of single-frequency collisional activation on mass/charge ratio and the number of ions in the ion trap.


Enantiomeric separations using bovine serum albumin immobilized on ion-exchange stationary phases.

Bovine serum albumin (BSA) can be readily immobilized on ion-exchange stationary phases by frontal analysis of a proper solution. This provides a simple means of adjusting the amount of BSA contained in the column and of measuring it accurately. Although the immobilization is ionic and not covalent, the columns are stable for extensive periods of time. If needed, they can be easily regenerated by the same frontal analysis procedure.


Theoretical and experimental investigation of steady-state voltammetry for quasi-reversible heterogeneous electron transfer on a mercury oblate spheroidal microelectrode.

A mercury microelectrode formed by electroreduction of mercury on an inlaid gold microdisk is experimentally shown to be well modeled by oblate spheroidal geometry when the ratio of the semiminor axis to the semimajor axis of the protruding drop is less than 1. The validity of the geometry is established by comparison of the experimentally determined coefficient in the steady-state diffusion current equation with the theoretical value for oblate spheroidal geometry. Spherical cap geometry is also shown to be an equally valid geometric model; however, theoretical treatment for this system is more difficult. The theory of a quasi-reversible electrode process is developed and applied to the determination of the electrode parameters of the RuIII(NH3)6/RuII(NH3)6 electrode reaction on a mercury oblate spheroidal microelectrode. Results agree well with others found in the literature for the same process on a mercury electrode.


Determination of lysergic acid diethylamide (LSD), iso-LSD, and N-demethyl-LSD in body fluids by gas chromatography/tandem mass spectrometry.

Procedures for detection and quantitation of lysergic acid diethylamide (LSD), iso-LSD, and N-demethyl-LSD by capillary chromatography/tandem mass spectrometry (GC/MS/MS) are presented. Several methods for derivatization, sample introduction, and ionization, in combination with mass spectrometry/mass spectrometry (MS/MS), have been evaluated for overall ionization efficiency and product-ion sensitivity and specificity. Fragmentation pathways derived from low-energy collision-induced dissociation (CID) spectra of protonated LSD, and the protonated trimethylsllyl derivatives of LSD (LSD-TMS) and deuterium-labeled analogs of LSD, have been proposed. Principal dissociations primarily involve the amide and piperidine-ring moieties in which losses of CH3 radical, CH3NH2, CH3NCH2, diethylamine, diethylformamide, and N,N-diethylpropenamide from MH+ are observed. Positive-ion ammonia chemical ionization and subsequent MS/MS analysis of the protonated molecules (MH+) of the trimethylsilyl (TMS) derivatives of LSD, iso-LSD, and N-demethyl-LSD provide a high degree of specificity for identification of these compounds in urine or blood at low-pg/mL concentrations. Negative-ion chemical ionization and GC/MS/MS analysis of the molecular anion (M-) of the trifluoroacetyl (TFA) derivative is well suited for trace-level identification of N-demethyl-LSD, a metabolite of LSD.


Matrix-assisted laser desorption mass spectrometry of proteins isolated by capillary zone electrophoresis.

A simple method for the off-line coupling of laser desorption mass spectrometry (LDMS) and capillary zone electrophoresis (CZE) is described. Representative mass spectra of subpicomole quantities of proteins isolated from CZE are presented and discussed. The current detection limit for bovine alpha-lactalbumin is 100 fmols injected onto the CZE column. Horse heart myoglobin was demonstrated to be stable in CHES/KCl, a CZE buffer, for at least 1 month, suggesting that some isolates can be safely stored for long time periods prior to LDMS analysis. Protein stability in 0.1% aqueous trifluoroacetic acid (TFA), a common solvent for LDMS, must also be considered. In the special case of porcine pepsinogen, significant (greater than 50%) degradation was observed within 5 min in TFA. In favorable cases, mass measurement accuracies of +/- 0.02% were obtained for protein isolates. Factors limiting mass measurement accuracy are presented. Finally, the possibility of identifying protein isolates, by combining N-terminal sequencing, molecular mass measurements, and selective peptide "mapping" procedures, is discussed.


Conformational effects in the reversed-phase chromatographic behavior of recombinant human growth hormone (rhGH) and N-methionyl recombinant human growth hormone (Met-hGH).

This paper examines the retention behavior of recombinant DNA-derived human growth hormone (rhGH) in reversed-phase chromatography and its separation from the closely related N-methionyl variant (Met-hGH). It is first shown that retention for rhGH decreases with increasing column temperature when 1-propanol (1-PrOH) is used as organic modifier. On the other hand, retention increases with temperature when acetonitrile (CH3CN) is employed. The differences in behavior for the two organic modifiers could be related to conformational changes in the protein as determined by solution and adsorption intrinsic fluorescence spectroscopy. Specifically, desorption and elution of rhGH using 1-PrOH could be correlated with a solvent-induced conformational change, with retention decreasing with increasing temperature due to the increasing ease of structural alteration. On the other hand for CH3CN the increase in retention correlated with temperature rise was related to a partial structural change yielding a more hydrophobic species. In this case, a surface-driven process is suggested. The work then turned to the separation of rhGH and Met-hGH where it was found for both organic modifiers optimum separation occurred at 45 degrees C and pH 6.5. Separate studies revealed that during the conformational change Met-hGH appeared more hydrophobic than rhGH since protein-protein aggregation was observed at a lower 1-PrOH concentration. It is suggested that this hydrophobic difference, which was optimized under the conditions cited above, resulted in the separation. The study demonstrates the importance of conformational changes in retention behavior and separation of protein samples.


Flow injection spectrophotometric determination of calcium, phosphate and chloride ions in milk.

Flow injection procedures for the determination of calcium, phosphate and chloride ions in milk samples are described. The reactions are based on the formation of coloured complexes and their spectrophotometric monitoring. A sample pre-treatment with acetate buffer was carried out owing to the complexity of the sample matrix. For chloride, a rapid and reliable automated procedure for direct measurement of its content in milk (using a dialyser to eliminate interferences) is also described. After optimizing the sample pre-treatment and flow injection variables, the procedures were applied to commercial milk; the results obtained agreed satisfactorily with those of the reference methods. With 50 mm3 samples, a working range of 0-15 ppm for calcium, 50-150 ppm for phosphate and 5-100 ppm for chloride is covered with a precision of better than 1.1%. The sample throughput was higher than 50 samples h-1. These preliminary experiments are the basis for the automation of the determination of calcium, phosphate and chloride ions using a computer-controlled, self-designed and laboratory-built autoanalyser.


Spectrofluorimetric flow injection method for the individual and successive determination of L-cysteine and L-cystine in pharmaceutical and urine samples.

A flow injection configuration is proposed for the determination of L-cysteine and L-cystine individually and for mixtures of both analytes. The procedure is based on the rapid oxidation of L-cysteine by thallium(III) with concomitant formation of fluorescent thallium(I). The inclusion of a selecting valve and of a copper-coated cadmium column in the configuration allows the successive determination of two analytes. Linear calibration graphs were obtained between 5 x 10(-6) and 5 x 10(-5) mol dm-3 of L-cysteine and between 2 x 10(-6) and 2 x 10(-5) mol dm-3 of L-cystine. The applicability of the method to the determination of L-cysteine and L-cystine in pharmaceutical preparations was demonstrated by investigating the effect of potential interferents and by the analysis of commercial preparations. The method was successfully applied to the determination of L-cysteine and L-cystine in urine samples.


Enzymic method for the spectrophotometric determination of choline in liquor.

A sensitive spectrophotometric method for the determination of choline in liquor is described. The method involves the conversion of choline into formaldehyde by sequential enzymic reactions (choline oxidase and catalase), followed by the formation of a chromophore with 4-aminopent-3-en-2-one. The calibration graph was linear in the range 0.4-15 micrograms cm-3 of choline. The relative standard deviation at 5 micrograms cm-3 of choline was 1.3%. There was no interference from most of the common ingredients of liquor. More than 95% of choline added at two levels was recovered from real samples. The method is simple, and the detection limit was 2 micrograms g-1 when 5 g of sample were assayed.


Behaviour of long-lived Chernobyl radionuclides in a soil-water system.

Field and laboratory experiments have been used to study the behaviour of long-lived radionuclides in the zone affected by the Chernobyl accident. Speciation of 90Sr and 137Cs in soils and bottom sediments was determined. The principal distinction of the Chernobyl fallout was that it contained a relatively small proportion of exchangeable forms because a considerable fraction of the radionuclides was incorporated as part of the insoluble fuel particles. Disintegration of fuel particles in soils and bottom sediments results in transition of non-exchangeable forms into exchangeable forms. Radionuclide species have different pathways and rates of migration in soils and bottom sediments. Migration of each chemical form was described by a convective-dispersive equation taking into account transformation processes of radionuclide species in soils or bottom sediments. Adsorption of 90Sr and 137Cs in the environment is controlled by the cation-exchange capacity and the selectivity of the solid phase (i.e., soil, bottom sediments and suspended matter) and the cationic composition of the liquid phase (i.e., soil solution, surface run-off and river or lake water). The corresponding parameters for the processes were obtained.


Cholesterol immobilization via ether-linked sepharose gels.

Cholesterol has been immobilized on Sepharose-6B via oxyether linkages to the 3- or 25-position. The 3- or 25-hydroxysterol methanesulfonates were coupled with epoxy-Sepharose-6B at 80 degrees C for 24 h. Approximately 2% of the ligand was incorporated into the gel. These types of affinity columns may be useful in purifying proteins that specifically bind or metabolize cholesterol.


Determination of blood lead in dried blood-spot specimens by Zeeman-effect background corrected atomic absorption spectrometry.

A simple procedure for the determination of blood-lead levels in dried blood-spot filter-paper specimens is described. A 3/16 inch dried blood spot was analysed by a method involving extraction of the lead into 1.25% (NH4)2HPO4-0.5% Triton X-100 solution. A matrix-based calibration was used for analysis, with use of a Zeeman-effect background corrected atomic absorption spectrometer. The within-run precision (% relative standard deviation) of the method at the low end of the analytical range was 19% at 0.36 mumol dm-3 and 14% at 0.60 mumol dm-3. The accuracy of the method was verified by recovery tests and by comparison with a routine whole-blood method. The mean blood lead level of 425 Toronto newborns was 0.19 mumol dm-3 (range 0-0.75 mumol dm-3).


Arsenic speciation by ion chromatography with inductively coupled plasma mass spectrometric detection.

Four As compounds were successfully separated and detected by single-column ion chromatography with inductively coupled plasma (ICP) mass spectrometric detection. The mass spectral interferent ArCl+ was reduced by chromatographically resolving chloride from the negatively charged arsenic species. Determination of four As species was investigated in urine, club soda and wine. Detection limits of 0.16 ng of As(III), 0.26 ng of As(v), 0.073 ng of dimethylarsinic acid (DMA) and 0.18 ng of methylarsonic acid (MMA) in wine were obtained. Sensitivity was further improved by using an He-Ar mixed gas ICP as the ionization source. However, the intensity of the ArCl+ interference was also increased using this plasma. Detection limits of 0.063 ng of As(III), 0.037 ng of As(v), 0.032 ng of DMA and 0.080 ng of MMA in club soda were achieved using the He-Ar plasma source. Similar limits of detection were found in urine and wine.


High-performance liquid chromatographic and derivative ultraviolet spectrophotometric determination of amoxycillin and dicloxacillin mixtures in capsules.

Two rapid assay procedures based on high-performance liquid chromatography (HPLC) and derivative ultraviolet (UV) spectrophotometry have been developed for the simultaneous determination of amoxycillin and dicloxacillin in two-component capsule formulations. The HPLC determination was carried out on a reversed-phase C8 column with use of a mobile phase consisting of methanol-0.02 mol dm-3 ammonium acetate (pH 5) (50 + 50) at a flow rate of 1.0 cm3 min-1, with UV detection at 230 nm. The fourth-derivative spectrophotometric procedure depends on the measurement of the derivative amplitudes, in 0.1 mol dm-3 NaOH, at 308.5 and 275 nm for amoxycillin and dicloxacillin, respectively. For both procedures, the calibration graphs were linear in the ranges 20-200 and 20-140 micrograms cm-3 for the HPLC and UV derivative methods, respectively, with an almost zero intercept and a correlation coefficient of 0.999. Commercial capsules and laboratory-prepared mixtures containing both penicillins in different proportions were assayed by the developed procedures. The results were of comparable accuracy as indicated by a statistical analysis of the data, using both t- and F-tests.


[The preservation of the whole corpse with natural color].

An especially low-odor embalming technique was developed over a 30-years-period using a total of 977 complete cadavers, numerous cadavers after autopsy, and in vitro series of fresh beef. The color, consistency, and transparency of the tissue were very well preserved. The technique met high standards of preservation without releasing harmful substances into the environment. Concentrations of formaldehyde in room air remained under the limit of detection by Dräger capillaries. The efficacy for disinfection of the method was confirmed by bacteriologic tests. None of the cadavers or samples developed molds.


[An arterial substance for subsequent injection during the preservation of the whole corpse].

Masses for arterial injection were evaluated in 360 whole cadavers that had been injected with preservation fluid. A mass consisting of dextrin, latex, and lead tetroxide produced the best results. This mass provided high radiopacity at large distances from the injection site and ideal firmness for dissection. The easy application of the mass makes it suitable for research purposes as well as for teaching dissections.


[Synovial recesses and bursae of the lumbosacral joint].

There are four types of cavities of the lumbosacral joints. Type A: The synovial membrane is all around attached to the margin of the articular facet of the superior articular process of the sacrum. Type B: The synovial membrane extends to the posterior surface of the sacrum forming a recess at the root of the superior articular process; this recess communicates widely with the cavity of the joint. Type C: The synovial membrane also forms a recess similar to that of type B, but the gap of communication is narrowed by a fibro-adipose meniscoid. Type D: The synovial membrane is attached in the same way as described in type A; besides a synovial bursa non communicating with the cavity is found at the root of the superior articular process. We have found 43% showing type A, 33% type B, 16% type C and 8% type D. The recesses and bursae described above enable the inferior articular process of the fifth lumbar vertebra to slide at the posterior surface of the sacrum, thus avoiding a painful rubbing during dorsiflexion of the lumbar spine. The existence of these sliding facilities does not depend on the range of the lumbosacral angle nor on the quality of the lumbosacral intervertebral disc.


[The olfactory region of the bat Scotophilus heathi. Light and electron microscopic studies].

The present investigation reports light and electron microscopical aspects of the main olfactory epithelium (MOE) of the insectivorous bat Scotophilus heathi. Serial frontal sections of the nose and associated structures reveal: 1) that the MOE is located on most of the ethmoturbinals and on the proximal upper portion of the nasal septum; and 2) that the vomeronasal organ is absent in this species. The ultrastructure of the MOE of Scotophilus heathi is similar to that observed in other vertebrates. Moreover, we did not observe significant morphological differences between the MOE of male and female animals. Nevertheless, the supra-nuclear region of the supporting cells of males and females shows a different amount and distribution of "lysosome-like" cell organelles at the two times of the year investigated.


Endothelial cell helix in small arterioles of human ureters. A study by scanning electron microscopy (SEM).

Microvessel corrosion casts of human ureters were prepared. Portions of casts were longitudinally cut by use of a pulsed Eximer laser beam in order to get insight into microvessel branches studied in SEM. Larger arterioles (greater than 30 microns in diameter) showed oval imprints of endothelial cell nuclei along the vessel axis. Imprints were found in furrows parallel to resin ridges. Smaller arterioles (less than microns in diameter) displayed imprints of nuclei aligned in a helix around the vessel axis. With regard to capillaries, one nucleus imprint was spaced far apart from the other. Venules demonstrated a reticular network of thick, attenuated resin ridges and of small meshes which consisted of round or oval nucleus imprints. It is concluded that smaller arterioles form an endothelial cells helix.


[The lateral articular surface of the medial cuneiform bone facing the intermediate cuneiform bone].

300 medial cuneiform bones were examined to explore the different forms of the joint surface on the lateral side of the bone facing the intermediate cuneiform bone. This joint surface consists of two parts, a proximal one and a dorsal one. The proximal part is orientated dorsoplantarally. At its proximal margin there exists a tight contact with the proximal joint surface for the navicular bone. The dorsal part is sited at the dorsal margin of the lateral side of the bone. Different forms of fusion of these part could be seen. In 62.67% they show tight confluence. In 17.67% the fusion is limited to a slight contact. In the rest, 19.67%, there exists no contact between these two parts. These forms correlate to the proximal height of the bone. A large proximal height extends the degree of separation of the dorsal joint surfaces (consisting of the dorsal section of the proximal part and the whole dorsal of the joint surface for the intermediate cuneiform bone as well as the joint surface for the second metatarsal bone). Therefore these joint surfaces are split to their components.


[The nomenclature of the blood vessels of the kidney].

The differing interpretations of the course of the renal artery-branches possibly are caused by the Nomina Anatomica. The course of the branches of the renal artery is described and based upon this less misleading names for these branches are suggested.


An anatomical investigation of the muscles of the pelvic outlet in Japanese giant salamander (Cryptobranchidae Megalobatrachus japonicus) with special reference to their nerve supply.

Four pelvic halves from two Japanese giant salamanders (Cryptobranchidae Megalobatrachus japonicus, one male and one female) were dissected in order to obtain detailed morphological data on the lumbosacral plexus and the muscles of the pelvic outlet. According to the positions of the passage of the metazonal nerves, the nerves can be divided into three groups: 1) the nerves passing dorsal to the caudofemoralis muscle, 2) the nerves passing between the caudofemoralis and the caudoischiadicus, 3) the nerves passing ventral to the caudoischiadicus. In consideration of the sites of origin from the lumbosacral plexus, a three-way stratificational analysis can be made, group 1 arising craniodorsal to group 2, and group 3 caudoventral to group 2. It is suggested that the three pelvic outlet muscles, the caudofemoralis, caudocruralis, and caudoischiadicus, have derived from the ventral muscles of the posterior limb, and have extended ventrocaudalwards.


Microvascular pattern during the growth of regenerating muscle fibers in the rat.

Quantitative methods were used to analyze changes in the capillarization of skeletal muscle during regeneration. The H-E staining technique was used to establish three arbitrary stages of fiber regeneration on the basis of qualitative histological evaluation. ATPase and light microscopic techniques were used to calculate capillary density, the number of capillaries surrounding each fiber, and the fiber area served by each capillary, for each of the three stages established. The results obtained indicated variations in the vascularization of regenerating fibers from one stage of maturity of another: capillarization was greater in the initial stages of regeneration, returning gradually to normal values by the final stages of the process. Interestingly enough the pattern in the capillarization of skeletal muscle during regeneration seems to be similar to that taking place during postnatal muscle development. Both processes appear to be governed by the increase in fiber size.


[The effect of cyclooxygenase inhibitors on the course of hereditary retinal dystrophy in RCS rats].

Daily administration of acetyl salicylic acid (ASA) and ibuprofen leads to an appreciable retardation in the process of retinal degeneration in the RCS rat which is dependent on the dosage given. The photoreceptor cell nuclei and inner segments are relatively well preserved in all regions of the retina. While the outer nuclear layer of 32 day old RCS rats is usually composed of only 3-4 rows, we found 8-9 nuclear rows exhibiting minimal pyknotic change in animals which had been treated with higher doses of ASA. These differences in layer thickness could be confirmed using morphometric analysis. The outer segments show evidence of degenerative change although they are in a clearly better condition than those found in untreated animals and in those animals treated at lower dose. Phagolysosomal structures which are not otherwise apparent in this strain of rat are detected only in the RPE cells of animals treated with higher dose. The animals treated with ibuprofen show essentially the same morphological changes although a corresponding effect in regard to the thickness of the outer nuclear layer was only achieved after a high dose. The determined dose for the optimal preservation (thickness) of the outer nuclear layer lies around 160 mg/kg body weight for acetyl salicylic acid and at 400 mg/kg body weight for ibuprofen.


The circulus arteriosus cerebri in the guinea pig.

Arteries to the brains of guinea pigs were injected with latex and the circulus arteriosus cerebri was studied. Most of the arterial blood to the guinea pig brain came from the internal ophthalmic and vertebral arteries. The internal carotid arteries were extremely thin. In addition to an anastomosis that was found between the right vertebral and the basilar arteries in one specimen, the right vertebral artery anastomosed with the left vertebral artery in eight specimens and in one animal the basilar artery was double along a major part of its length.


The connective tissue sheath of uterus and vagina in the human female fetus.

A morphological study concerning the development and arrangement of the connective tissue surrounding uterus and vagina was performed by investigating epoxy resin impregnated sections through the pelves of 9-37-week-old female fetuses, of newborn children and a three-year-old child. In order to study the histology of the connective tissue in detail paraffin, semithin and ultrathin sections were used. The organization of the pelvic connective tissue in female fetuses can best be studied in middle-aged fetuses (19-28 weeks of gestation). We here report that no supportive ligaments of the uterus are found in fetuses of that age group, but that the connective tissue covering uterus and vagina is part of the connective tissue sheaths of neighbouring structures. The fetal uterus is interposed between dense subperitoneal connective tissue circularly covering the vesico- and rectouterine pouch. Dorsally the subperitoneal tissue of the rectouterine pouch joins the circularly arranged connective tissue of the rectal adventitia. A triangularly-shaped plate of dense connective tissue is situated at each lateral border of the cervix uteri. Dorsally it is continuous with the circularly running fibres of the rectouterine pouch. Whereas the dorsal wall of the vagina is loosely connected with the rectal adventitia, fibrous connective tissue intimately fuses the muscular wall of the vagina with that of the neck of the bladder and of the urethra. No evidence is provided for the existence of smooth muscle cells and elastic material within the connective tissue covering the fetal uterus.


The sperm penetration test (P-test) can predict fecundability in the male partner from infertile couples.

Three hundred and twenty-one consecutive couples were investigated for infertility at Hvidovre University Hospital in the period from November 1977 to June 1985. The male partners were evaluated in two ways: the classical semen analysis, and the ability of sperm to penetrate fresh hen egg white, the P-test. A Cox regression analysis was used to describe the relation between these variables and fecundability, i.e. the time required to conceive. Four of thirteen variables--the number of morphologically normal spermatozoa, the number of motile spermatozoa, the P-test, and the man's age--each have significant relation to the fecundability. However, when covariation is considered, only the P-test and the man's age possess significant prognostic information, whereas the variables of the classical semen analysis do not. This indicates that the P-test may replace the classical semen analysis when trying to predict individual pregnancy probabilities. Finally, the P-test and the man's age are combined to form a prognostic index which predicts the fecundability of the male partner in the individual infertile couple.


Use of an ATP-supplemented medium for the conservation of human semen and the effect of caffeine on the motility of preserved sperm. Results in artificial insemination.

Human semen can be preserved at 4 degrees C in an artificial medium containing 15% ATP-MgCl2 for up to 5 d. This method has been applied to samples from subjects with either normal or abnormal semen. After the conservation period the recovery of sperm motility observed was good when low doses of caffeine (3.5 or 7.0 mM) were added.


Intra-uterine insemination with prepared sperm vs. unprepared first split ejaculates. A randomized study.

In this randomized prospective study, we determined the conception rate following intra-uterine insemination with washed and prepared sperm, or with the first portion of a split ejaculate, in couples with longstanding male (n = 27, 70 treatment cycles) or cervical infertility (n = 14, 29 treatment cycles). Folliculogenesis and ovulation were induced by human menopausal gonadotropin and human chorionic gonadotropin. Significantly more couples conceived in the male infertility group following intra-uterine insemination with washed sperm, than after intra-uterine insemination with split ejaculate (9 vs. 2; P less than 0.05), while no difference in pregnancy rate (2 vs. 2) was found by the two intra-uterine insemination methods in the cervical infertility group.


Evolutionary genetics of ruminant lysozymes.

Comparative studies of mammalian lysozymes and their genes have contributed to knowledge of how new functions arise during evolution. The recruitment of lysozymes for functioning in the stomach fluid of ruminants has occurred in response to selection pressures that are partly known and on a time-scale that is known. A semiquantitative analysis of adaptive evolution is thus made possible by the ruminant lysozyme system. Large-scale production of lysozyme by the stomach lining entailed gene duplication as well as a change in gene expression. Remoulding of the lysozyme for working and lasting in the stomach fluid involved accelerated amino acid replacements, which may have been facilitated by intergenic recombination. The possibility that multigene families can accelerate adaptive evolution, by virtue of their capacity for bringing together functionally coupled substitutions, receives emphasis in this review.


Bivariate flow cytometry of farm animal chromosomes: a potential tool for gene mapping.

Bivariate flow karyotypes of chromosomes from sheep, cattle and pig lymphocytes and from a cattle-mouse somatic cell hybrid line were obtained using a dual laser fluorescence-activated cell sorter (FACS). Pig chromosomes were resolved into 19-20 peaks, indicating that most, if not all, pig chromosomes could be separated by this technique. Sheep chromosomes showed incomplete separation but three clear peaks, presumably representing the three large metacentric chromosomes, plus five other clusters were obtained. Cattle chromosomes showed poor separation but about four peaks could be distinguished, indicating that certain chromosomes could be sorted in this species. The use of cattle-mouse hybrids may enable other individual cattle chromosomes to be obtained. It is concluded that FACS separation will be a useful additional tool for gene mapping.


Effect of different types of textile fabric on spermatogenesis. I. Electrostatic potentials generated on surface of human scrotum by wearing different types of fabric.

This paper comprises a study of the electrostatic potentials generated on the surface of the scrotal area when different types of textile fabric were worn. 21 healthy volunteers were divided into 3 equal groups. The first group was dressed in underpants made of 100% polyester, the second of 100% cotton, and the third of a 50:50% polyester:cotton mixture. With an electrostatic kilovoltmeter, the electrostatic potentials were measured 1 h after wearing the pants once at daytime and a second time at night. The test was repeated 4 times, each on a separate day. No electrostatic potentials were detected on the cotton underpants. The polyester pants showed the highest potentials (mean 338.9 +/- 25 SD Volt cm-2) while the mixed polyester:cotton pants produced less than half that level (mean 148.3 +/- 16 SD Volt cm-2). The readings at daytime were higher than at night, probably due to the higher temperature during the day. The accumulated electrostatic charges on the pants are due to the friction of the pants with the skin. As a result of friction, equal and opposite charges are accumulated on the skin surface and on the inner surface of the pants facing the skin. Eventually, an 'electrostatic field' is produced traversing the scrotal contents and may disturb the testicles and/or epididymis leading to disordered spermatogenesis.


Loss of acid phosphatase from rat spermatozoa as a method for assessing the acrosome reaction.

A method is presented for evaluating the extent of the acrosome reaction by measuring the release of acrosomal acid phosphatase from rat spermatozoa during incubation under capacitating conditions. Treatment of spermatozoa with lysophosphatidylcholine or Triton X-100 released the acid phosphatase from the sperm cell. Using this enzymatic method we could not detect an alteration in enzyme activity following 5 h incubation under capacitating conditions. The effect of in vitro capacitation for 5 h in the absence or presence of heparin or ionophore A23187 was studied. Incubation in the presence of heparin (10 micrograms ml-1) caused a 32% increase in enzyme activity. After exposure of the spermatozoa to ionophore A23187 (0.5 microM) 16% increase of enzyme activity could be detected.


Identification of the major histocompatibility complex in the ring-necked pheasant, Phasianus colchicus.

Reciprocal immunization between parents of an Illinois family of ring-necked pheasants, Phasianus colchicus, resulted in antisera detecting two pairs of alloantigens segregating among 13 progeny. The four alloantigens were tentatively designated as 1 and 2, transmitted antithetically by the sire, and 3 and 4, transmitted antithetically by the dam. Genetic segregation occurring in second-generation progeny demonstrated that these two pairs of antigens belonged to a single genetic system. This alloantigen system was shown to correspond serologically to the major histocompatibility complex (MHC) of the chicken by cross-reactivity of the antigens of this system with specific subregional chicken MHC reagents, appropriately absorbed with erythrocytes of individual pheasants. These four haplotypes of the pheasant MHC were subsequently designated as MhcPhco-B1, MhcPhco-B2, MhcPhco-B3 and MhcPhco-B4. Traditional immunogenetic analysis of 30 pheasant families produced in this study disclosed a minimum of 14 pheasant haplotypes of this alloantigen system (MHC) to be segregating in the population under evaluation.


Numerical variation of nucleolar organizer regions after silver staining in domestic and wild Suidae (Mammalia).

Selective silver staining was used to investigate the cellular distribution of numbers of nucleolar organizer regions (NORs) in domestic pigs (Sus scrofa) of eight different breeds, the European wild boar (S. scrofa scrofa), Indonesian wild boar (S. scrofa vittatus), Javan warty pig (S. verrucosus), Sulawesi warty pig (S. celebensis), and pigmy hog (S. salvanius). In the domestic pig as well as in the wild (sub)species of Sus, actively transcribing ribosomal RNA genes were found to be present in the secondary constrictions of chromosome pairs 10 and 8. Chromosomes 10 were consistently Ag-positive. Chromosomes 8 less frequently showed Ag-NORs, resulting in different mean numbers of Ag-NORs per individual animal. Mean Ag-NOR numbers per breed or (sub)species were generally higher in the wild representatives of Sus than in the domestic breeds. The highest mean numbers of Ag-NORs were observed in the Meishan breed and in S. celebensis and S. salvanius. The Meishan breed appears to be conservative in Ag-NOR staining pattern, being more comparable to the Asian wild Suidae than to the European breeds.


The effect of clomiphene citrate and its Zu or En isomers on the reproductive system of the immature male rat.

The effect of clomiphene citrate (CC) and of its Zuclomiphene (ZuC) and Enclomiphene (EnC) isomers on the reproductive organs of immature male rats under different experimental conditions is reported. CC, ZuC, and EnC were administered daily to groups of either intact or castrated rats between the age of 21-44 d. This led to inhibition of weight increase of testis and accessory glands in the intact group. Spermatogenesis was arrested at the stage of primary spermatocyte following CC and ZuC treatment, and at the stage of young spermatids by EnC treatment. In the castrated group clomiphenes significantly stimulated weight increase of seminal vesicles (SV) compared with castrated control animals, but the former group were unable to achieve organ weight gain comparable to that in normal controls. Administration of human Chorionic Gonadotropin (hCG) together with CC or each of its isomers to intact animals, abolished the drug effect on spermatogenesis and on reproductive organ growth. Administration of CC, ZuC, and EnC together with testosterone to castrated animals, abolished the drug effect on growth inhibition of accessory glands. In intact treated rats LH and testosterone secretion were suppressed by all forms of clomiphenes. In the castrated group ZuC proved to be the most potent inhibitor of LH secretion. Therefore, it is inferred that ZuC and EnC have different potencies as far as their biological activity in the immature male rat is expressed.


Bovine epididymal sperm proacrosin-acrosin system: quantification and partial characterization.

Several studies suggest that acrosin, an acrosomal trypsin-like serine proteinase, plays a role in fertilization. The enzyme is present in an enzymatically inactive precursor form, called proacrosin and is believed to be converted to the enzymatically active form(s) through one/multiple physiological event(s) prior to the sperm penetration of the zona pellucida. Although, the proacrosin-acrosin system of several species has been well documented, the study of the enzyme system in bovine caput and cauda epididymis (where the maturation of spermatozoa occurs) has not been characterized. The present study demonstrates the quantification and partial characterization of the proacrosin-acrosin proteinase system in unpurified acrosomal extracts of bovine caput and cauda epididymal sperm. Proacrosin activation followed the sigmoidal type of activation curve. Activation experiments demonstrate that almost 80-90% of this protein exists in zymogen (proacrosin) form either in ejaculated or caput and cauda epididymal spermatozoa. Time-course activation studies showed that the zymogen in isolated spermatozoa was completely converted to active non-zymogen form in 3 and 5 h after removal from the cauda and caput regions, respectively, at pH 8.0 at 25 degrees C. This conversion was markedly inhibited by calcium in a dose dependent manner and the inhibition was reversible. On the other hand, calcium has a stimulatory effect on the hydrolytic activity of acrosin. These studies reveal that the proacrosin-acrosin system can be identified in crude extracts of bull epididymal and ejaculated sperm.


Cloning and sequencing of the porcine lactoferrin cDNA.

cDNA clones encoding the entire porcine lactoferrin protein were isolated and sequenced. The porcine lactoferrin cDNA sequence presented here is 2259bp in length and encodes a leader peptide of 19 amino acids and a mature protein of 684 amino acids. Comparisons with other lactoferrins indicate a single glycosylation site. The iron- and anion-binding sites, and the cysteine residues involved in disulphide bonds, are conserved between the lactoferrin proteins.


Sequence of porcine beta-lactoglobulin cDNA.

cDNAs encoding porcine beta-lactoglobulin were isolated and sequenced. The porcine beta-lactoglobulin cDNA is 768bp in length and encodes a pre-protein of 178 amino acids. One additional cDNA clone was found to encode an additional amino acid (lysine) in the mature protein.


Assignment of the porcine tumour necrosis factor alpha and beta genes to the chromosome region 7p11-q11 by in situ hybridization.

The loci of the porcine tumour necrosis factor genes, alpha (TNFA) and beta (TNFB), have been chromosomally assigned by radioactive in situ hybridization. The genomic probes for TNFA and TNFB yielded signals above 7p11-q11, a region that has been shown earlier to carry the porcine major histocompatibility locus (SLA). These mapping data along with preliminary molecular studies suggest a genomic organization of the SLA that is similar to that of human and murine major histocompatibility complexes.


Demonstration of serum albumin (ALB) polymorphism in wild rabbits, Oryctolagus cuniculus, by means of isoelectric focusing.

Genetic polymorphism of serum albumin was demonstrated by isoelectric focusing in wild rabbit populations from Portugal and England. Gene frequencies were estimated to be (1) ALB*1 = 0.47, ALB*2 = 0.49, ALB*3 = 0.04, in Portugal, and (2) ALB*1 = 0.60 and ALB*2 = 0.40, in England. One hundred Portuguese domestic rabbits of mixed breeds were all of ALB 1 type.


A new genetic variant Z2 in the Pi system of horses.

A new genetic variant in the horse Pi system, designated Z2, was reported in Polish Arabs by using two-dimensional agarose polyacrylamide gel electrophoresis. The frequencies of Pi alleles F, G, L, L2, N, S, U, W, Z and Z2 were found to be 0.036, 0.005, 0.171, 0.013, 0.008, 0.237, 0.416, 0.003, 0.107 and 0.004 respectively.


The sequence of porcine alpha s1-casein cDNA: evidence for protein variants generated by altered RNA splicing.

A cDNA library was constructed from mRNA isolated from lactating porcine mammary gland and screened with a bovine alpha s1-casein cDNA clone. Three classes of cDNA isolated varied in the number of bases within the coding region. The full length porcine alpha s1-casein cDNA is 1124bp and codes a preprotein of 206 amino acids. The other two classes of alpha s1-casein cDNA lacked 18bp and 60bp respectively when compared to the 1124-bp cDNA sequence. PCR amplification confirmed the presence of these sequences in total RNA. These differences appear to be due to altered RNA splicing.


Detection of leucochimaerism in bovine twins by DNA fingerprinting.

Karyotyping and hypervariable genetic markers indicate extensive leucochimaerism between pairs of dizygotic twins in cattle, a result of placental vascular anastomosis. The extent of this chimaerism includes both kind and number of cells exchanged. All heterosexual twin pairs harboured two types of leucocytes, having either XX or XY chromosome pairs, and 30 of 31 pairs of twins shared identical DNA fingerprints. Although chromosome results from skin fibroblasts indicate that some chimaerism occurs in the skin, the low level allows for differentiation of genotypes between twins. The results warrant against the common practice of using blood samples for DNA typing if twinning is not properly documented.


Genetic polymorphism of plasminogen and vitamin D binding protein in red deer (Cervus elaphus L.).

Genetic polymorphism was detected in the red deer (Cervus elaphus L.), plasma proteins, plasminogen (PLG) and vitamin D binding protein (GC) using antiserum to human proteins. The affinity of the antisera to deer plasma was less than 10% that of a human standard but they bound specifically to proteins of molecular weight expected for GC and PLG. Three codominant alleles of GC and five of PLG were observed. In a set 124 farmed deer calves and their parents, six calves had genotypes which were not consistent with the expectations of inheritance. Further inconsistencies were found when variation in isocitrate dehydrogenase (IDH) and transferrin (TRF) was examined. Using genetic models which included pedigree error parameters the data were shown to be consistent with genetic inheritance of all loci in a data set containing approximately 4.8% (SE 1.4%) parent-progeny pedigree mismatches. In samples from four deer populations representative of the red deer introduced to New Zealand the GC and PLG polymorphisms provided a probability of paternity exclusion (PE) of between 0.34 and 0.54 and when IDH and TRF were also included the PE was between 0.46 and 0.66. The four populations differed significantly in allele frequency, which supports historical evidence that they originate from separate introductions of small numbers of European red deer.


Canine plasma alkaline phosphatase polymorphism and its relationship with the canine Tr blood group system.

A canine blood group antigen, QN, which was detected by a naturally occurring alloantibody in the antiglobulin test, was shown to be antigenically related to the human A and cattle J antigens by absorption experiments. Family studies supported a dominant mode of inheritance with the gene controlling the production of QN being dominant to the gene responsible for its absence. Frequencies of the QN and Tr antigens and serological data strongly suggested that the two antigens are identical. Two canine plasma alkaline phosphatase variants, F and S, were detected by starch gel electrophoresis, pH 8.65. Assuming genetic control of two codominant alleles, ALPF and ALPS, the distributions of types in families differed significantly from expectation. A relationship between the ALP and Tr(QN) systems was demonstrated with Tr-positive animals having a significant deficiency of S alkaline phosphatase types.


Sheep blood polymorphism and genetic divergence between French Rambouillet and Spanish Merino: role of genetic drift.

Rambouillet sheep originating from Spanish Merino have been maintained in France as a small and closed flock since their importation. After 190 years of independent evolution, the flock has markedly differentiated from its Spanish parental population. The observed differences between them were characterized by the fixation in Rambouillet of the Mb and F30 alleles, which occurred in Spanish Merino with frequencies of 0.90 and 0.80 respectively (at two distinct blood group loci M and F30) and by the absence in Rambouillet of other alleles or phenogroups (at the Tf and the A, B, C blood group loci) which were observed in Spanish Merino with frequencies ranging from 0.10 to 0.28. On the basis of their phenotypic distributions at 11 blood polymorphic loci, the two populations differed significantly from each other (total chi 2 values = 352.62, 23 df, P less than 0.001). By comparing the observed magnitude of gene frequency differences between Rambouillet and Spanish Merino with the estimate of inbreeding coefficient for Rambouillet obtained from pedigrees, it appeared that the observed genetic differences could be attributed to the evolutionary change due to random drift in the small and closed flock of Rambouillet.


Detection of multiple beta-casein (CASB) alleles by amplification created restriction sites (ACRS).

Direct sequencing of polymerase chain reaction (PCR) amplified DNA has been used to detect the DNA sequences for bovine beta-casein (CASB) A3 and B variants. Based on these sequences we have designed primers which create allele-specific restriction sites in the PCR product. Restriction analysis of PCR product generated in one reaction enable us to identify the A1, A2, A3 and B alleles of CASB rapidly without the use of radioactivity.


Genetic analysis of fingerprints in Mérinos d'Arles x Booroola Merino crossbred sheep.

The M13.13 minisatellite probe, consisting of a polymer of the M13 VNTR consensus sequence, cross-hybridized to ovine DNA and allowed detection of several polymorphic loci. Individual specific patterns were obtained in sheep using this probe. Pedigree analysis showed that individuals were heterozygous for most of the DNA fragments detected (88%). By studying the segregation of male's variable DNA fragments, a minimum of 10 loci were defined. The ovine DNA 'fingerprint' obtained with M13.13 is polymorphic enough to be used efficiently in animal identification, paternity testing, and possibly as a source of genetic markers for linkage analysis.


Expression and characterization of ovine major histocompatibility complex class II (OLA-DR) genes.

Previous work made use of nucleic acid probes corresponding to different subtypes of the class II regions of the human and murine major histocompatibility complex (MHC) to isolate seven different alpha and 24 different beta genes of the ovine MHC from two cosmid libraries. In an attempt to identify pairs of alpha and beta genes capable of cell surface expression, all permutations of alpha and beta genes were in turn transfected into mouse L-cells. Two pairs of alpha and beta genes co-expressed and stable ovine MHC class II L-cell lines were developed. The expressed alpha genes had previously been defined as DR-alpha homologues (DRA) by differential Southern hybridization to human subtype specific class II probes. The expressed ovine beta genes were also assigned as ovine DR-beta homologues (DRB) on the basis of their sequence having a higher degree of similarity with human DRB than any other subtype. A total of eight out of 23 anti-sheep class II specific monoclonal antibodies were typed OLA-DR specific by FACScan analysis using the L-cell lines.


Cloning and sequencing of the porcine kappa-casein cDNA.

cDNA clones encoding porcine kappa-casein were isolated and sequenced. The porcine kappa-casein cDNA is 851 bp in length and encodes a preprotein of 188 amino acids.


The sequence of porcine alpha s2-casein cDNA.

cDNA clones encoding the entire porcine alpha s2-casein message were isolated and sequenced. The porcine alpha s2-casein cDNA is 1093 bp, excluding the poly(A) tail, in length and encodes a preprotein of 235 amino acids.


The sequence of porcine beta-casein cDNA.

Porcine cDNAs clones encoding beta-casein were isolated and sequenced. The porcine beta-casein cDNA is 1100bp in length, excluding the poly(A) tail, and encodes a preprotein of 232 amino acids.


Genetic polymorphism in sheep plasma detected using antibodies to human plasminogen.

Protein variation was identified in sheep when Western blots of polyacrylamide gels (routinely used to resolve transferrin polymorphism) were stained using antibodies to human plasminogen. The affinity of the antibodies to ovine plasma was less than 7% that of a human standard but they bound specifically to a single polymorphic protein. In 146 lambs and their parents the inheritance of the ovine plasminogen antigen polymorphism was consistent with four autosomal alleles segregating codominantly. However, an additional two lambs had types which were incompatible with their putative parents. The pedigrees of these lambs were tested by DNA fingerprinting and shown to have been incorrectly recorded. The genetic polymorphism detected by human plasminogen antiserum provided a probability of sire exclusion (PE) ranging from 0.04 to 0.32 and a polymorphic information content (PIC) of 0.08 to 0.50 in flocks of five sheep breeds: Perendale, Romney, Merino, Texel and Coopworth (in order of increasing genetic variation in this locus). Significant differences in allele frequency were observed between breeds but sampling did not assess the variation among flocks within a breed.


[Clonidine premedication and isoflurane anesthesia to reduce bleeding in otologic surgery].

Seventy-seven ASA 1 patients scheduled for ear surgery were premedicated orally, 90 min before anaesthesia. They were randomly assigned to two groups, according to the drug used: hydroxyzine alone (group T, n = 39) or combined with clonidine (4.9 +/- 0.3 micrograms.kg-1) (group C, n = 38). Anaesthesia was induced with midazolam (0.3 mg.kg-1) and alfentanil (30 micrograms.kg-1). Ventilation was controlled with a 50/50 v/v mixture of oxygen and nitrous oxide (FETCO2 = 4 to 4.5%), and anaesthesia was maintained with repeated injections of alfentanil (15 micrograms.kg-1 at the start of surgery, and then every 15 min) and with isoflurane (mean end-expiratory concentration 0.6 +/- 0.3 vol %). Surgical bleeding was assessed every ten minutes on a numerical scale with four values. A bloodless surgical field was obtained by adjusting the isoflurane concentration up to 2 MAC, and by using a trinitrine infusion as required. Cardiovascular monitoring included an electrocardioscope and automatic blood pressure measurements. Before induction of anaesthesia, the blood pressure was lower in group C (84.7 +/- 11.2 vs. 95.9 +/- 106 mmHg) (p less than 0.001); the difference in heart rate was not significant (65 +/- 15 vs. 70.6 +/- 14 b.min-1). Moderate stable intraoperative hypotension was obtained in both groups. However, mean arterial blood pressure (C:65.8 +/- 7.8 mmHg; T: 73 +/- 9.4 mmHg) and heart rate (C: 53.4 +/- 6.8 b.min-1; T: 60.4 +/- 8 b.min-1) were significantly lower in the patients premedicated with clonidine (p less than 0.001). There were more periods of sinus bradycardia (heart rate less than or equal to 50 b.min-1), mostly seen before the beginning of surgery, in group C patients (p less than 0.01); atropine was also required more often (when the heart rate was less than or equal to 40 b.min-1) in this group of patients (NS). The comparative assessment of surgical field quality was in favour of group C (no troublesome bleeding) as opposed to the control group (16% troublesome bleeding); there were also more bloodless surgical fields in the former group (73.7% vs. 48.7% in group T, p less than 0.05). This study therefore demonstrated that clonidine premedication before anaesthesia with isoflurane was helpful in decreasing bleeding during ear surgery.


[EMLA analgesic cream for venopuncture during anesthetic induction in children].

A prospective study was designed to assess the quality of skin analgesia provided by the EMLA anaesthetic cream, an eutectic mixture of local anaesthetics (prilocaine and lidocaine). The children, aged 3 months to 15 years, and scheduled for genital and urinary surgery, were allocated to two groups, those aged less than or equal to 5 years (35 +/- 14 months, n = 17), and those aged greater than 5 years (97 +/- 26 months, n = 22). The cream (1.6 +/- 0.6 g) was applied by a nurse in the ward as a thick layer on the area of skin to be anaesthetised (on the dorsum of the hand and at the elbow), and covered by a closed adhesive dressing. This required 2.6 +/- 1.7 min, and was considered to be very easy (72%) or easy (28%). The venepuncture (22 or 20 gauge catheter) was carried out by one of the seven anaesthetists of the paediatric surgical units, 92 +/- 51 min after the cream had been applied. The children aged less than or equal to 5 years complained of pain of intensity 7.5 +/- 2.2 (CHEOPS scale, range 4 to 13) and, for those aged greater than 5 years, 24 +/- 21 on a visual analogic scale (range 0 to 100). Local adverse effects occurred in ten patients (skin paleness, erythema, or both). It was concluded that EMLA cream provides convenient analgesia for venepuncture in toddlers and children.


[Pharmacokinetics of methohexital given by constant rate intravenous infusion].

The pharmacokinetic characteristics of a constant rate methohexitone infusion were studied in young ASA 1 patients undergoing maxillofacial surgery. They were randomly assigned to two groups; group M patients (n = 7) were given 9 mg.kg-1.h-1 of methohexitone for one hour, and group MF patients (n = 7) 9 mg.kg-1.h-1 of methohexitone with 7 micrograms.kg-1.h-1 of fentanyl, also for one hour. Blood samples for determining methohexitone concentrations were obtained at various times, from before the start of the methohexitone infusion up to 19 h afterwards. In twelve patients, a two-compartment model was appropriate to characterize the decrease of methohexitone concentration; for the other two (one in each group), a three-compartment model was applied. There were no statistically significant differences between the two groups. Elimination half-life in group M was 3.22 +/- 1.96 h, and total plasma clearance 8.54 +/- 2.8 ml.kg-1.min-1. The wide variations in pharmacokinetic parameters between subjects may explain some unpredictable variations in duration of action of methohexitone. Fentanyl did not modify methohexitone pharmacokinetics, which remained of the first order. However, it potentiated the barbiturate's action: extubation was only possible after stopping the infusion for 39.4 min +/- 22 min in group MF, and 15.4 min +/- 6 min in group M (p less than 0.01). At that time, plasma concentrations were respectively 3.12 +/- 0.99 mg.l-1 (group MF) and 5.71 +/- 2.09 mg.l-1 (group M), (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


[Caudal block in children: analgesia and respiratory effect of the combination bupivacaine-fentanyl].

A study of the duration of analgesia and of the respiratory response to hypercapnia was carried out in 14 children who had had a caudal block with either bupivacaine alone (group B) or combined with fentanyl (Group B+F). Fourteen ASA I or II 5 to 10-year-old children undergoing genital and urinary surgery were included. They were not premedicated. At first, general anaesthesia was induced with halothane and nitrous oxide in oxygen. Thereafter, caudal anaesthesia was then carried out with 1 ml.kg-1 of 0.25% bupivacaine with adrenaline 1 in 200,000. Group B+F patients were also given 1 microgram.kg-1 of fentanyl in 1 ml of normal saline, and those in Group B 1 ml of normal saline. The level of sensory loss on leaving the operating theatre as well as the duration of motor paralysis were monitored. Postoperative pain was scored with Hannalah and Broadman's score (0 to 10) 2, 4, 8 and 24 h after the caudal block. Respiratory rate (fR), tidal volume (VT) and minute ventilation (VE) were assessed 10 min before induction of general anaesthesia, and 30, 60 and 120 min after the caudal anaesthesia. Petco2 was also measured before induction of general anaesthesia, and 60 and 120 min after caudal anaesthesia; at the same times, the ventilatory response to hypercapnia was assessed using Read's method with a Douglas bag containing 7% CO2 and 93% O2.(ABSTRACT TRUNCATED AT 250 WORDS)


[Effect of hygrophobic filter or heated humidifier on peroperative hypothermia].

A study was carried out to find out whether the use of a hygrophobic filter (Pall, Ultipor) or of a heated humidifier (Dräger, Aquapor) during surgery had any effect on a patient's intraoperative core temperature and thermal balance. Seventy-five ASA I or II patients scheduled for gynaecological surgery were randomly assigned to three groups: group A (n = 25), where no warming device was used; and two groups (n = 25 for each) where inhaled gases were humidified and heated with either a hygrophobic filter set up between the endotracheal tube and the Y-piece (group B) or a heated humidifier set to 100% saturation at a temperature of 41.5 degrees C (group C). The patients were all anaesthetised with the same technique (thiopentone 5 mg.kg-1, dextromoramide 0.03 mg.kg-1 and 0.1 mg.kg-1 of either pancuronium or vecuronium, followed by enflurane with nitrous oxide in oxygen); the perfused fluids were not heated. Room, tympanic, rectal, oesophageal and four skin (thorax, arm, leg, thigh) temperatures were measured with calibrated Exacon thermistances, on arrival in the operating theatre, during induction, every ten minutes for two hours, and then every twenty minutes for two hours more. Ramanathan's and Burton's formulae were used to calculate mean skin temperature and heat loss respectively. In the recovery room, patients were warmed up with an electric blanket. Shivering was ranked from "0" to "+ +". There were no differences between groups as far as age, drug doses, perfusion volumes and room temperature were concerned.(ABSTRACT TRUNCATED AT 250 WORDS)


[Antibioprophylaxis in pulmonary surgery: a randomized trial with cefamandole versus placebo].

A prospective double-blind study was designed to assess the efficacy of antibiotic prophylaxis in lung surgery. It included 114 patients undergoing lung surgery for primary or secondary malignant tumours randomly assigned to two groups. Group A patients (n = 59) were given cefamandole intravenously every four hours, three times, starting from induction of anaesthesia. The dose was determined according to the patient's weight: 1.5 g for patients weighing less than 60 kg, 2.5 g for those weighing between 60 and 80 kg, and 3 g for those above 80 kg. Group B patients (n = 55) were given a placebo at the same times. Nineteen other patients were excluded because either the tumour was found to be infected, or the patient had to be mechanically ventilated postoperatively, or an exploratory thoracotomy only was carried out, or they were allergic to beta-lactam antibiotics. The efficacy of antibiotic prophylaxis was assessed by recording the incidence of postoperative infections, the length of the patient's stay in hospital, and the need to use an antibiotic treatment. Patients, their sputum and wound were examined every day, and their temperature recorded. The white blood cell count and chest X-ray was carried out every day for the first week. All the drain and catheter tips were cultured, as well as sputum and blood (every three days). In case of infection, samples were obtained and cultured. Both groups of patients were similar with regard to age, risk factors (smoking habit, diabetes mellitus), and type of surgery (segmentectomy, lobectomy, pneumonectomy). There were 9 postoperative infections in group A, and 22 in group B (p = 0.003).(ABSTRACT TRUNCATED AT 250 WORDS)


[Thrombosis of the superior vena cava after prolonged catheterization. Treatment by progressive removal of the catheter combined with urokinase-heparin administration].

A retrospective study of 13 cases of complete superior vena cava thrombosis due to prolonged catheterization is reported. All the polyurethane catheters had been inserted by anaesthetists in theatre between January 1985 and December 1989, using Seldinger's technique. On the 10th day after the first catheter had been placed, the catheter was replaced by using a guide wire. Superior vena cava obstruction was diagnosed clinically (eyelid, neck, facial and upper limb oedema, cyanosis, collateral venous circulation, lack of blood reflux in the catheter). The clinical diagnosis was confirmed by phlebography. Treatment consisted in administration of urokinase (1,500 IU.kg-1.h-1), together with 10,000 to 25,000 IU.day-1 of heparin, both being injected via the catheter. The catheter was removed progressively (1 cm every 2 to 4 h). As soon as a venous pressure curve was obtained, the catheter was completely removed. Overall 8,784 catheters were inserted during that period. The incidence of superior vena caval obstruction may therefore be estimated to be 1.48 per 1,000 catheters. The catheters remained in place up to a mean of 14.75 +/- 9.6 days before the obstruction occurred. Treatment with urokinase and heparin lasted a mean of 2.53 +/- 1 days. The clinical picture returned to normal in all 13 patients. Phlebography carried out in three of them, after treatment, showed an excellent degree of venous repermeability. Thrombolysis was confirmed by the increase in the concentration of D-dimers, without any decrease in fibrinogen concentration. There were five haemorrhagic complications, including two haematomas of the psoas muscle, one of which required surgical drainage.(ABSTRACT TRUNCATED AT 250 WORDS)


[Effects of diazepam and incidence of hypoxemia during acute chloroquine poisoning].

The effects of diazepam and the incidence of hypoxaemia on the course of acute chloroquine poisoning were studied prospectively in 21 patients. Were excluded patients who had ingested more than one drug or who had major symptoms on admission (systolic blood pressure less than 80 mmHg; QRS greater than 0.12 s; cardiac dysrhythmias, respiratory disturbances). Arterial blood gases were measured on admission (T0) and 15 min after 0.5 mg.kg-1 of diazepam had been given (T1). Gastric lavage was carried out as soon as the results of the blood gases had been obtained, and after treatment of hypoxaemia (PaO2 less than 90 mmHg). An infusion of diazepam (1 mg.kg-1.day-1) was then given. Arterial blood gases were measured after 1 (T2), 6 (T3), 12 (T4) and 24 h (T5). Hypoxaemia was present on admission in four patients who had a PaO2 = 75 +/- 10 mmHg (Pa(sys) = 130 +/- 19 mmHg; blood chloroquine concentration = 8.2 +/- 5.2 mumol.l-1; kaliemia = 3.1 +/- 0.3 mmol.l-1; PaCO2 = 35 +/- 1 mmHg). In two patients, hypoxaemia decreased after the initial dose of diazepam (T1); however, oxygen was still required by the other two at that time. Oxygen was no longer needed by any patient at T2, as all the blood gas values had returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS)


[Effects of diazepam on mortality from acute chloroquine poisoning].

A retrospective study was carried out, over a twelve-year period, of all cases of acute chloroquine poisoning where more than 2 g of chloroquine had been taken. It included 386 patients; of these, 60 who had taken drugs other than chloroquine, and 17 who had ingested less than 1 g of the drug, were excluded. The remaining 309 patients were allocated to two groups: a "control group", consisting of the patients admitted between January 1973 and April 1980 (n = 146), and a "diazepam group", made up of those admitted from May 1980 to December 1989 (n = 163). The patients in the latter group had had the same symptomatic treatment as those in the control group, and had been routinely given a 0.5 mg.kg-1 bolus of diazepam on admission followed by 0.1 mg.kg-1.day-1 for every 100 mg of chloroquine supposed to have been ingested. Both groups were divided into three subgroups, those patients with cardiorespiratory arrest, and those with, and those without, symptoms on admission. No statistically significant difference was found between either the control and diazepam groups or between subgroups, concerning the distribution of age, sex, amount of chloroquine supposed to have been ingested, delay in hospital admission and death rate. However, there was a higher death rate in the asymptomatic subgroup not treated with diazepam than in the diazepam group. Therefore, the routine use of diazepam for the treatment of acute chloroquine poisoning does not seem to be justified in symptomatic cases and in those with inaugural cardiac arrest.(ABSTRACT TRUNCATED AT 250 WORDS)


[Prostate transurethral resection syndrome].

The "transurethral resection of prostate" syndrome (TURPS) is the clinical manifestation of the resorption into the patient's body of a large amount of glycocolle-containing irrigating fluid used for this procedure. The full clinical picture, which is seldom seen, consists of dyspnoea, nausea, arterial hypertension, an increased central venous pressure, cerebral oedema, cardiogenic shock and renal failure. Improved surgical techniques, as well as incomplete and atypical forms of the syndrome could explain this low incidence. Absorption into the blood stream may be rapid, by way of the prostatic venous plexi, or slower, from the spaces around the prostate and under the peritoneum. The present-day pathophysiological theory explains this syndrome by an acute hyponatraemia, sometimes dissociated from the hypoosmolality, the toxicity of glycocolle, and the neurological effects of hyperammonemia. Acute hyponatraemia, with blood sodium concentrations below 115 to 120 mmol.l-1, should be considered as potentially serious. The different mechanisms involved may act alone or together, thus explaining that the minor forms of the syndrome mostly consist of a neurological picture. The emergency treatment depends on the natraemia. It includes diuretics and progressive reloading of the patient with sodium in case of severe hyponatraemia with seizures. The best prevention is a correct surgical indication and technique. The resection should not last for more than 90 to 120 min. The major problem remains the early diagnosis of TURPS. Carrying out this surgery under regional anaesthesia is helpful for this purpose, but, in the near future, the best means might be the monitoring of expired ethanol concentrations.


[Cardiac tamponade and central venous catheterization].

A 32-year-old female patient was operated on for a residual colonic stricture occurring after hemicolectomy. A right internal jugular central venous catheter was inserted during the anaesthetic for postoperative parenteral feeding. The anaesthetic combined both general and epidural anaesthesia, the latter being continued for postoperative analgesia (10 ml.h-1 of 0.125% bupivacaine). Two days later, the patient complained of sudden chest pain, with restlessness, tachycardia, cyanosis, resulting in ventricular tachycardia and cardiac arrest. When admitted to the surgical intensive care unit, the patient was in deep coma and had nonsustained ventricular tachycardia, a left haemopneumothorax and a pneumopericardium. The patient died before a definitive diagnosis was made. Postmortem examination revealed an ulcerated anterior pillar of the tricuspid valve, as well as a perforation of the right ventricle and a communication between the pericardium and the left pleural cavity. The diagnosis and treatment of this rare life-threatening complication may be very difficult. It prevention consists in using short catheters for internal jugular venous access, and checking the tip's position radiologically by opacifying the catheter.


[Evaluation of 5 years of experimental in utero surgery for the repair of diaphragmatic hernia].

The incidence of diaphragmatic hernia in he newborn is 1:2,500 to 5,000 births. An extensive American programme of foetal surgery for in utero repair of this defect has shown that the pulmonary hypoplasia was due to compression of lung tissue by the herniated organs. This process could be interrupted be repairing the diaphragmatic defect early enough in utero (before the 28th week). The results of five years of experimental surgery for in utero correction of diaphragmatic hernia are presented here. The experimental model used was the Macaca fascicularis monkey, the pregnant female of which having several gestational and endocrine characteristics similar to those of the pregnant woman. Three different experimental programmes were carried out successively. A first series including five animals was used to experiment both surgical and anaesthetic techniques; three foetuses died in utero. The second series (10 animals) was designed to find a suitable protocol for tocolysis. One female died after surgery, and seven other foetuses also died. The third group (13 animals) was the main study group. The diaphragmatic hernia was first created surgically, and then repaired. No foetus died from the surgery. One female died before giving birth, and one offspring died shortly after birth as its mother had no milk. After the encouraging results obtained with this last series of animals, the procedure was applied to human cases, with the Hospital Ethical Committee's approval. In the first case, the foetus died postoperatively as a result of the rupture of the diaphragmatic prosthesis.(ABSTRACT TRUNCATED AT 250 WORDS)


[Acute respiratory insufficiency caused by diffuse pulmonary hemorrhage].

Two cases of adult respiratory distress syndrome due to diffuse pulmonary haemorrhage are reported. The first patient was treated with azathioprine, prednisolone, cyclosporine and ranitidine for haemorrhagic rectocolitis; the second has untreated primary biliary cirrhosis. Haemoptysis only occurred in the latter. Both had severe isolated hypoxaemia. Chest X-rays revealed bilateral alveolar infiltrates. Bronchoscopies showed a diffusely bleeding bronchial tree. Both patients recovered after having been mechanically ventilated with positive end-expiratory pressure for six and eight days respectively. The cause of the diffuse pulmonary haemorrhage was, in the first case, severe thrombocytopaenia (17,000 G.1-1) of central origin, and, in the other patient, an unspecified vasculitis. Diffuse pulmonary haemorrhage should be added to the list of possible causes of the adult respiratory distress syndrome.


[Acute respiratory insufficiency in sickle cell disease].

Two cases are reported of acute respiratory failure occurring during sickling crises. In the first one, the crisis was characterised by priapism, and in the other one, by abdominal pain. The different causes of these respiratory effects are discussed: infection, fat embolism, pulmonary infarct, haemodynamic pulmonary oedema, as was probably the case in the first patient, or non haemodynamic pulmonary oedema due to sickling, as during conventional treatment of a sickling crisis (oxygen, antibiotics, blood transfusion, cytapheresis). Invasive investigations may contribute to keeping up the clinical picture, because of hypoxic sickling. The water equilibrium of these patients must be monitored with great care. Worsening of the patient's condition despite 48 h of correct treatment must lead to the search for a specific cause.


[Treatment of algodystrophy of the knee: diagnostic error and misplacement of the peridural catheter].

A fifty-year-old female patient, with a history of reflex sympathetic dystrophy of the left hand and right ankle, complained of pain in her right knee. The skin was slightly oedematous and red, whereas the knee X-ray was quite normal. These findings were thought to be related to another episode of reflex sympathetic dystrophy, and treatment with continuous epidural morphine and lidocaine was prescribed. Catheter insertion was uneventful. However, the lack of pain relief led to the suspicion of femoral neuralgia. Plain X-ray films of the lumbar spine showed the epidural catheter to be passing through the L2-3 foramen. The catheter was removed. The development of fever and major inflammatory signs of the knee revealed gout arthritis. This case stresses the need for careful repeated clinical examination in order to make the right diagnosis. On the other hand, when the expected effect of drugs administered by the epidural route fails to appear, the catheter's position should be promptly checked by X-ray.


[Paraplegia after spinal anesthesia].

A case of paraplegia occurring after a spinal anaesthetic is reported. The 79-year-old man was admitted for a fractured neck of femur. Twenty years previously, he had had pharyngeal surgery and a tracheostomy. He had also undergone a prostatectomy for prostate cancer, and had been on oestrogen therapy for two years. He complained of dyspnoea at rest and his chest film showed diffuse pulmonary opacities. In order to avoid possible intubation and respiratory complications, spinal anaesthesia was performed without any problems in the L4 space. After the surgery, the patient recovered all his motor and sensory functions in the lower limbs. On the second postoperative day, he suffered from a motor paralysis of the right leg, which spread to the left leg on the fourth day. NMR imaging showed several vertebral metastases, together with anterior and lateral epidural invasion responsible for cord compression. Treatment with tetracosactide was begun, but the patient died six weeks later in his home, not having recovered any neurological function at all in his lower limbs. In fact, it was only after the procedure that the anaesthetist was informed that, at the time the prostate cancer had been diagnosed, vertebral body metastases, of which the patient had not been informed, were already present. The part played by the spinal anaesthetic in the occurrence of the paraplegia is not clear. It is reminded that such a technique should be used with extreme care in patients having a neoplasm with a very often high incidence of vertebral metastases.


[Physical modeling of inhalation anesthesia].

A new physical model simulating the pharmacokinetics of volatile anaesthetics is presented. It consists in a ventilator connected to a water manometer. Gas is removed from the gaseous part of this manometer with a constant rate pump. This gas flow is directed thereafter into three capacitances with valves and pumps: one capacitance only contains air, representing the lungs, and the other two olive oil, representing the visceral and muscle compartments. Halothane, enflurane and isoflurane (1 vol%) were administered to this model with different values of cardiac output and alveolar ventilation. There was good concordance between the values of FA/FI that were measured in this model and those calculated by computer simulation. No correction factor was required. Such a physical model may therefore be used to test new techniques of administration of volatile agents.


[Venous echo Doppler: a future standard test in the diagnosis of thrombosis of the lower limbs?].

Duplex ultrasound imaging provides both an ultrasound picture of the tissue being explored and the sound of the flow speed in a particular area. Colour Doppler imaging gives colour codes for the different flow rates. Venous pathology is always investigated, with ultrasound, on both sides, at the iliac and caval, femoral, and foreleg levels. For the latter, excellent equipment and patient position are required. Ultrasound criteria of deep venous thrombosis (DVT) are the incompressibility of the vein being investigated, and detection of the thrombosis with the ultrasound probe. Doppler criteria are asymmetric spontaneous flows, the loss of their respiratory rhythm, and the lack of flow induced by contraction of muscle. The colour Doppler draws provides the contours of a partial or floating thrombus. Compared with phlebography, ultrasound imaging has a sensitivity and specificity of 96%. Colour Doppler imaging is even more accurate: both sensitivity and specificity increase to 100%. The limitations of ultrasound are: its dependence on the investigator, the difficulty of making the diagnosis of recurring DVT when there already are previous sequellae, obesity, intra-abdominal gas, skin lesions or inflammatory oedema. Phlebography remains necessary in 5% of cases. The advantages of this technique are its non invasiveness, excellent diagnostic performance, low cost, and the possibility of diagnosing another cause, whether responsible for the clinical signs or not. It is indicated for the diagnosis of DVT, routine screening in patients at risk, and the early and late follow-up of patients. Today, ultrasound imaging should always be carried out before other investigations.(ABSTRACT TRUNCATED AT 250 WORDS)


[Accidental spinal anesthesia in obstetrics. Limits of epidural test-dose].

Two cases of accidental spinal anaesthesia occurring in obstetrical patients are reported and discussed. Epidural anaesthesia had been asked for by the women, both being free from any significant medical history. A Tuohy needle was inserted in the midline between L3 and L4 with the patient sitting. The extradural space was identified by the loss of resistance using saline. The test-dose (2 ml and 4 ml of 1% lignocaine respectively) was administered five minutes before changing the patient to the supine position. In the first case, after a test-dose had remained without any effect, 8 ml of 0.25% bupivacaine were injected, about one hour later. The patient rapidly complained of paralysis of her legs and difficulties in breathing. Her blood pressure decreased from 120/80 mmHg to 90/60 mmHg. The upper level of analgesia reached T4. She improved after infusion of 1.51 of lactated Ringer's solution. Endotracheal intubation was not required. Delivery was assisted with a vacuum extractor. In the other patient, when the extradural space had been located, there was a small reflux of clear fluid which did not contain any glucose. As the test-dose did not result in any effect. 2 ml of 2% lignocaine with adrenaline were injected. This was followed by an immediate sensory loss in the legs, extending up to T10. Caesarean section was decided on, without any further injections (foetal macrosomia, breech presentation). Both patients totally recovered without any sequela. Both children had an Apgar score of 10 at 1 and 5 minutes. The type of test-dose is discussed. Smaller volumes of more concentrated solutions are recommended.


[Abscess of the atrium: a complication of central venous catheters].

A 27-year-old female patient was admitted for parenteral feeding and treatment of an acute episode of her steroid dependent Crohn's disease. An intravenous catheter was inserted, the tip being located in the right atrium. This central line was accidentally removed ten days later and replaced with another one also ending in the right atrium. The patient's condition improved over a four-week period. Thereafter, she suddenly became confused and complained of abdominal pain with contracture. She displayed renal failure, coagulation disorders, and decreased blood cell counts. This was followed by a septic shock requiring mechanical ventilation and adrenaline. Laparotomy failed to reveal a cause for the patient's condition. She improved and was extubated four days later. From then on, she had fever (39 degrees C) again. Her catheter was replaced, the tip of this third one being inside the superior vena cava. Staphylococcus epidermidis was obtained from the catheter tip, as well as from four consecutive blood cultures. Despite administration of three different antibiotics, the high fever persisted (40 degrees C). Finally, the occurrence of a systolic murmur led to the discovery, with cardiac ultrasonography, of a mobile right atrial abscess, which engaged into the tricuspid valve at every contraction. Surgery to remove this mass was rapidly carried out. The bacterium was the same as that which had been isolated from the catheter. The patient improved and was able to return home a fortnight later. The diagnosis and prevention of this complication is discussed. The tip of central venous catheter should not be kept inside the atrium.


[Clinical study of EMLA analgesic cream in pediatric locoregional anesthesia].

The advantage of EMLA cream for regional blocks (spinal or caudal blocks) was assessed in 46 children. The study included three groups: group 1, with 1 to 6 month-old children (n = 11); group 2, with children aged between 6 months and 5 years (n = 21), scheduled for emergency surgery and with full stomach; group 3, with 5 to 10 year-old children (n = 14), who were to have a regional block as an alternative to general anaesthesia. The cream was applied as a thick layer on the area of skin to be anaesthetised and covered by a closed adhesive dressing, approximately 2.2 h before performing the regional block. A dose of 1 to 2 g was used in children of more than 1 year, and 0.5 to 1 g for those of less than 1 year. All the blocks were carried out by the same anaesthetist. The technique was considered as being little (45/46) or not constraining (1/46). Patient cooperation in carrying out the regional block was judged to be good or very good in 34/46 children. Additional sedation was required in 7 children of groups 2 and 3. The block was as easy to carry out as usual in 37/46 children. In all three groups, most children complained of little or no pain. Local adverse effects occurred in 19/46 patients, consisting of erythema (14), skin paleness (4), or both (1). The investigator qualified the use of EMLA cream as satisfactory or very satisfactory in 36/46. It is concluded that EMLA cream provides convenient analgesia for regional blocks in toddlers and children.


[Tracheal intubation in patients with cervical spine injuries using a fiber optic laryngoscope].

Eleven patients, with a cervical spine injury and scheduled for elective cervical spine fusion at least 48 h after their initial trauma, were intubated using a new fiberoptic laryngoscope (Bullard). This technique uses either a semi-rigid guide independent of the laryngoscope blade, or a rigid one attached to the blade. The cervical spine was immobilized with either a collar or a halo. General anaesthesia was carried out with thiopentone, fentanyl and vecuronium bromide. Orotracheal intubation was successful at the first attempts in 10 out of the 11 patients. No mobilization of the cervical spine occurred. In the first six patients, the semi-rigid guide was used, and the rigid one in the remaining five. The anaesthetist who carried out the intubations was always the same. Using the rigid guide was easier than the semi-rigid one. This is confirmed by the time required, 44 +/- 22 sec for the rigid guide, and 97 +/- 92 sec for the semi-rigid one. In the patient in whom this technique failed at the first attempt, endotracheal intubation was carried out by the nasal route and controlled by the fiberoptic laryngoscope. This technique enables a rapid and easy orotracheal intubation in trauma patients with an immobilized cervical spine, but careful training is necessary.


Fibrinogen and viscosity as risk factors for subsequent cardiovascular events in stroke survivors.

To investigate whether abnormalities in blood viscosity predict a poor prognosis for subsequent cardiovascular events in stroke survivors. Nested case-control study among a cohort of survivors of a first stroke, followed for an average of 2 years. Patients with a second stroke, myocardial infarction, or cardiovascular death were matched with patients who did not have such events (control patients). Buchberg-Klinik, Bad Tölz, Germany, a specialized center for stroke rehabilitation. A total of 625 consecutive patients. Twenty-one patients (3.5%) were lost to follow-up. Sixty pairs were matched. Native and hematocrit-standardized blood viscosity at three shear rates, hematocrit, plasma viscosity, fibrinogen, erythrocyte sedimentation rate, total leukocyte count, and the matching variables. Eighty-five patients had a second stroke, myocardial infarction, or died due to a cardiovascular event. Patients with re-events had higher blood viscosity and fibrinogen levels than the control patients. In the 60 matched pairs, the mean of the paired differences between patients with re-events and control patients was 5.03 mPa.s (95% CI, 1.262 to 8.941; P = 0.01) for native blood viscosity at shear rate 0.7 s-1, for plasma viscosity, 0.044 mPa.s (CI, 0.006 to 0.083; P greater than 0.02), and for fibrinogen, 0.056 g/L (CI, 0.010 to 0.101; P greater than 0.02). Odds ratios were significantly increased only for plasma viscosity (odds ratio, 2.86; CI, 1.06 to 8.43) and fibrinogen (odds ratio, 3.67; CI, 1.31 to 11.69). Hyperfibrinogenemia is an independent risk factor for cardiovascular events in stroke survivors. Intervention trials with fibrinogen lowering measures may be warranted.


A low-molecular-weight heparinoid compared with unfractionated heparin in the prevention of deep vein thrombosis in patients with acute ischemic stroke. A randomized, double-blind study.

To compare the relative safety and efficacy of a low-molecular-weight heparinoid (ORG 10172) with unfractionated heparin in the prevention of deep vein thrombosis in patients with acute ischemic stroke. Double-blind randomized trial. Seven Canadian university-affiliated hospitals. Eighty-seven patients with acute ischemic stroke resulting in lower-limb paresis. Patients received either low-molecular-weight heparinoid, 750 anti-factor Xa units twice daily, or unfractionated heparin, 5000 units subcutaneously twice daily. Treatment was continued for 14 days or until hospital discharge if sooner. Deep vein thrombosis was diagnosed using 125I-labeled fibrinogen leg scanning and impedance plethysmography. Venography was indicated if either test was positive. Overt hemorrhage, major or minor, was assessed clinically. Venous thrombosis occurred in four patients (9%) given low-molecular-weight heparinoid and in 13 patients (31%) given heparin (relative risk reduction, 71%; 95% CI, 16% to 93%. The corresponding rates for proximal vein thrombosis were 4% and 12%, respectively (relative risk reduction, 63%; P greater than 0.2). The incidence of hemorrhage was 2% in both groups. Low-molecular-weight heparinoid, given in a fixed dose of 750 anti-factor Xa units subcutaneously twice daily, is more effective than subcutaneous low-dose heparin for the prevention of deep vein thrombosis in patients with acute ischemic stroke.


Comparison of the specificity and sensitivity of Hemoccult and HemoQuant in screening for colorectal neoplasia.

To compare the Hemoccult II and HemoQuant tests regarding their specificity and sensitivity in screening for colorectal neoplasia. Cross-sectional study in which subjects underwent the two tests in parallel, after excluding dietary hemes and peroxidase-rich foods. HemoQuant results were analyzed for three different upper limits of normality (1.5, 2.0, 3.0 mg/g feces). A university hospital and the surrounding community. A total of 150 healthy volunteers, 124 patients with colorectal cancer, and 86 patients with adenoma. Blinded comparison of the specificity and the sensitivity of the two tests for colorectal cancer and adenoma. Test specificity was 99.3% with Hemoccult and was 92.7%, 94.7%, and 97.3% with HemoQuant, depending on the cutoff point; differences between Hemoccult and HemoQuant were significant when cutoffs of 1.5 and 2.0 mg/g were used in HemoQuant testing (6.6% [95% CI, 2.3 to 11.1] and 4.7% [CI, 0.8 to 8.5], respectively). Test sensitivity for colorectal cancer at all sites was 89.5% with Hemoccult and was 83.1%, 74.2% and 62.9% with HemoQuant, for the 1.5, 2.0, and 3.0 mg/g cutoffs, respectively; differences were significant with the 2.0 and 3.0 mg/g cutoff points (6.4% [CI, 6.7 to 24.0] and 26.6% [CI, 17.4 to 35.9%], respectively). The two tests had similar levels of sensitivity for cancers proximal to the splenic flexure, but sensitivity was substantially lower with HemoQuant for the more distal cancers. For all adenomas, test sensitivity was 30.2% with Hemoccult and ranged from 45.4% to 22.1% with HemoQuant. Although HemoQuant provides a precise measurement of fecal heme and its porphyrin degradation products, the test's performance characteristics in the detection of colorectal neoplasia are less satisfactory than those of Hemoccult II, a qualitative test for the presence of heme.


Return to work after uncomplicated myocardial infarction: a trial of practice guidelines in the community.

To evaluate the effectiveness of practice guidelines for return to work after acute myocardial infarction when disseminated from a university-based setting to a practice-based setting. Randomized clinical trial. A total of 187 patients with uncomplicated acute myocardial infarction. Patients were randomly assigned to the intervention (n = 95) or to usual care (n = 92). The intervention consisted of a treadmill test, a counseling session based on the test results, and a consultation letter from a cardiologist to the primary care physician. Individualized recommendations for the timing of return to work, contained in the consultation letter, were based on the patient's risk for recurrent cardiac events. Questionnaire, chart review, and a phone interview documented the timing of return to work and the rates of cardiac death, coronary angioplasty, coronary artery surgery, and recurrent myocardial infarction. Median intervals between acute myocardial infarction and return to work were similar in both groups (intervention, 54 days; usual care, 67 days; P greater than 0.2). Among patients without myocardial ischemia, however, the interval was shorter in the intervention group than in the usual care group (38 days compared with 65 days, respectively, P = 0.008). Among patients with myocardial ischemia, intervals were similar in both groups (80 days compared with 76 days, respectively, P greater than 0.2). Practice guidelines developed in a university-based setting were not as successful in hastening return to work after uncomplicated acute myocardial infarction when tested in a practice-based setting. Physicians' reluctance to follow guidelines for patients with myocardial ischemia reflected their concern with prognosis even though medical outcome was good.


Trimethoprim-sulfamethoxazole compared with vancomycin for the treatment of Staphylococcus aureus infection.

To compare trimethoprim-sulfamethoxazole (TMP-SMZ) and vancomycin regarding efficacy and safety in the therapy of serious Staphylococcus aureus infections. Randomized, double-blind comparative trial. A tertiary-care hospital. One hundred and one intravenous drug users hospitalized with S. aureus infection. Cure and failure rates; blood and wound cultures; minimum inhibitory and bactericidal concentrations; serum inhibitory and bactericidal titers; temperature; leukocyte count; durations of treatment and hospitalization; and toxicity. Of 228 intravenous drug users, 101 had S. aureus infection and were included in the efficacy analysis (43 received TMP-SMZ and 58 received vancomycin). Methicillin-resistant S. aureus (MRSA) accounted for 47% of S. aureus isolates, and 65% of patients were bacteremic. Infections were cured in 57 of 58 vancomycin recipients and in 37 of 43 TMP-SMZ recipients (P less than 0.02). Failure occurred mostly in patients with tricuspid valve endocarditis and only in those with infection caused by methicillin-sensitive S. aureus (MSSA). The mean duration of bacteremia was 6.7 days in TMP-SMZ recipients and 4.3 days in vancomycin recipients. Among 222 subjects hospitalized for at least 24 hours, toxicity rates were similar for TMP-SMZ (23%) and vancomycin (20%) recipients; nausea and vomiting were associated with TMP-SMZ and inflammation at the intravenous site was associated with vancomycin. Forty-four percent of TMP-SMZ recipients and 29% of vancomycin recipients experienced side effects in the efficacy cohort (P greater than 0.05). Vancomycin is superior to TMP-SMZ in efficacy and safety when treating intravenous drug users who have staphylococcal infections. However, all treatment failures occurred in patients with MSSA infection at any site. Therefore, TMP-SMZ may be considered as an alternative to vancomycin in selected cases of MRSA infection.


Autocrine secretion--10 years later.

The concept of autocrine secretion, its subsequent modifications, its application for understanding pathogenesis of disease, and its potential for developing new approaches to prevention and treatment are reviewed. Peptide growth factors (cytokines) act as local autocrine and paracrine mediators of tissue homeostasis. Many diseases, including cancer, atherosclerosis, rheumatoid arthritis, and other fibrotic diseases characterized by chronic inflammation, are associated with aberrant expression and cellular coordination of the homeostatic action of these regulatory molecules. Modern biotechnology and pharmacology offer unique opportunities for the therapeutic prevention and treatment of these molecular and cellular lesions, using either cytokines or other agents that modify their synthesis and activity.


NIH conference. Respiratory disease in the immunosuppressed patient.

Pulmonary complications, both infectious and noninfectious, are an important cause of morbidity in patients with various types of immunosuppression. The appropriate response to these clinical problems requires an understanding of pulmonary host defense and of the various types of systemic immunosuppression. Infectious and noninfectious pulmonary complications may vary according to the type of immunosuppression as well as to the degree and duration of immunosuppression. Appropriate clinical management also requires an understanding of the clinical problems commonly seen in specific groups of immunosuppressed patients and an understanding of the sensitivity, specificity, and potential complications associated with the available diagnostic approaches to those patients. Because respiratory disease in these patient groups may progress rapidly to respiratory failure, an expeditious evaluation based on the knowledge of likely causes of respiratory disease and prompt specific or empiric therapy are indicated. Specific sets of algorithms for the evaluation of both focal and diffuse pulmonary disease may facilitate such an evaluation. In addition, an aggressive approach to the prevention of pulmonary disease including immunization, prophylaxis, and immunomodulation (for example, colony stimulating factors) may be warranted in specific subgroups at risk.


The influence of hyperglycemia on outcome of cerebral infarction.

To determine whether hyperglycemia in the acute stroke period is associated with worse survival and functional outcome after accounting for acute stress response and chronic hyperglycemia. Prospective, county-wide, multicenter cohort study. A community hospital, a university hospital, and a Veterans Affairs hospital. A cohort of 146 patients hospitalized with new atherothrombotic stroke. Admission blood glucose concentration, demographic characteristics of patients, 24-hour urinary catecholamine, serum cortisol, and glycosylated hemoglobin levels; outcomes included mortality and functional outcome (Barthel index and Fugl-Meyer score) at 5, 30, 90, and 180 days after stroke. Of the 996 patients with possible acute stroke who were screened, 146 (15%) were eligible for and consented to participate in the study; in most cases, exclusion from study was based on the absence of acute, atherothrombotic stroke. Overall, no evidence was found of a significant univariate association between admission blood glucose level and survival (relative risk, 1.02; 95% Cl, 0.94 to 1.09) or functional outcome (univariate regression coefficient for adjusted Fugl-Meyer score at day 30, - 0.36; Cl, - 1.08 to 0.27). This absence of an association persisted after adjustment for significant predictors of outcome in a multivariate model. These data do not support an association between level of glycemia and outcome from acute stroke.