Rapid and simple method for the determination of alpha 1-acid glycoprotein in serum by column liquid chromatography. A rapid and simple method for the determination of alpha 1-acid glycoprotein (AAG) in serum was developed by using an anion-exchange column for clean-up of serum and a hydroxyapatite column for high-performance liquid chromatography (HPLC). A good correlation was observed between this HPLC method and the conventional radial immunodiffusion method. The method may also be used to determine the AAG concentration in the serum of experimental animals.


Improved procedure for the drying and storage of polyacrylamide slab gels. An improved procedure for the drying and storage of polyacrylamide (PAA) slab gels of various sizes and acrylamide contents is presented. A PAA slab gel is preferentially fixed in methanol-acetic acid-glycerol-water (1.0:1.0:0.1:2.0) and then dried under low pressure in an adapted photoprint dryer between Cellophane and polyethylene foils at 75-80 degrees C for 50-90 min. Strong, transparent and flexible sheets are obtained that can be stored for extended periods. The potential of the procedure is exemplified following electrophoresis of both lipopolysaccharides and protein isolated from Coxiella burnetii bacterium.


High-performance liquid chromatographic monitoring of metabolic products resulting from the treatment of mouse hepatoma cells with N6-cycloalkylated nucleosides. High-performance liquid chromatography was used to analyze cell lysates and growth medium of mouse hepatoma cells, separately treated with N6-cyclopropyl-, N6-cyclobutyl-, and N6-cyclopentyladenosines, in an effort to gain insight into the mechanism by which these modified nucleosides exert their cytotoxic effect(s). The corresponding 5'-monophosphate of the respective modified nucleoside was detected in the separate cell lysate samples. Both the modified nucleoside and its corresponding 5'-monophosphate were detected in the separate growth medium samples and their relative concentrations therein were determined. These results indicate that the cytotoxicity of these N6-cycloalkylated nucleosides may be attributed to their 5'-monophosphates within the cells.


Simple high-performance liquid chromatographic method for the determination of captopril in biological fluids. A rapid, simple and sensitive column-switching high-performance liquid chromatographic procedure for the determination of captopril in plasma and urine had been developed. p-Bromophenacyl bromide was used as a derivatizing reagent to react with captopril to form a product that showed ultraviolet-absorbing properties. For plasma samples the protein was removed with 6% perchloric acid before injection. The urine samples were directly injected into the chromatograph. The column-switching system was equipped with a pre-column (5 cm x 0.5 cm I.D.) packed with muBondapak C18 (37-50 microns) and an analytical column (15 cm x 0.5 cm I.D.) packed with YWG-C18, 10 microns. Impurities were washed from the pre-column with 0.2% acetic acid and the retained substances were eluted into the analytical column with acetonitrile-water-acetic acid (35:65:0.4, v/v). Captopril was detected at 260 nm. The calibration curve was linear in the range 20-1000 ng/ml for plasma and 10-200 micrograms/ml for urine. The recoveries averaged 103.2 and 99.5% for plasma and urine, respectively. The coefficients of variation were all less than 10%.


Improved high-performance liquid chromatographic determination of ciprofloxacin and its metabolites in human specimens. A simple approach to the quantitation of ciprofloxacin and its three metabolites, M1 (desethylene-ciprofloxacin), M2 (sulfo-ciprofloxacin) and M3 (oxo-ciprofloxacin), in human serum, urine, saliva and sputum is described. This assay allows the parent drug and its metabolites to elute and be resolved in a single chromatogram at 280 nm using a linear gradient. The procedure involved liquid-liquid extraction. Separation was achieved on a C18 reversed-phase column. The limit of detection of ciprofloxacin is 0.05 microgram/ml and that of its three metabolites is 0.25 microgram/ml. This method is sufficiently sensitive for pharmacokinetic studies.


Automated high-performance liquid chromatographic method for the determination of mianserin in plasma using electrochemical detection. An automated high-performance liquid chromatographic method for the determination of mianserin in plasma is described. Extraction and injection of the samples were automatically done by the Gilson ASPEC system using C8, 100-mg Supelclean solid-phase extraction columns. The extracts were chromatographed on a reversed-phase C18 column (150 mm x 3.9 mm I.D.) with a phosphate buffer-acetonitrile-methanol mobile phase and the analytes detected electrochemically. Calibration curves were linear to at least 53.7 ng/ml at which the between-day relative standard deviation was 5% and the recovery 101%. The limit of quantification was 1.67 ng/ml at which the between-day relative standard deviation was 9% and the recovery 92% using a sample volume of 0.5 ml. The method was applied to the determination of mianserin in the plasma of normal human volunteers participating in a comparative bioavailability study.


Determination of D-alanine and D-glutamic acid in biological samples by coupled-column chromatography using beta-cyclodextrin as mobile phase additive. A high-performance liquid chromatographic assay for monitoring traces of D-alanine and D-glutamic acid in peptide and protein hydrolysates is presented, which uses a two-column set-up with column switching. The main advantage of the proposed method is the higher reliability of the results, compared with the analysis by derivatization and single-column chromatography of the diastereomers. A non-chiral alkyl-silica reversed-phase column is combined with a second column, in which beta-cyclodextrin is used as a chiral mobile phase additive. The amino acids of the hydrolysate are dansylated, and the amino acid of interest is separated from the others on the first column and transferred to the second column where chiral resolution is performed. The transfer volume of ca. 200 microliters is small enough not to cause any peak distortion or dilation in the second column.


Determination of free amino acid enantiomers in rat brain and serum by high-performance liquid chromatography after derivatization with N-tert.-butyloxycarbonyl-L-cysteine and o-phthaldialdehyde. The concurrent determination of free amino acid enantiomers and non-chiral amino acids in rat brain and serum was accomplished by high-performance liquid chromatography with fluorimetric detection after derivatization with N-tert.-butyloxycarbonyl-L-cysteine and o-phthaldialdehyde. The method revealed the presence of a large amount of free D-serine (0.22 mumol/g of tissue; D/D + L ratio = 0.25) in the brain whereas D-aspartate and D-alanine were established to be at trace levels. These results further support the presence of D-serine in adult brain tissues as demonstrated by recent work using gas chromatography.


Quantitative analysis of collagen and elastin cross-links using a single-column system. The separation of both the immature and mature cross-links present in collagen together with the stable cross-links of elastin has been achieved on a single ion-exchange column. This technique avoids the current necessity for two different systems, ion-exchange and high-performance liquid chromatography-fluorescence techniques. The value of the method is illustrated by the comparison of the cross-link contents of aging bovine skin, from foetal to old age.


High-performance liquid chromatographic determination of dihydroorotate dehydrogenase of Plasmodium falciparum and effects of antimalarials on enzyme activity. A reversed-phase high-performance liquid chromatographic technique for the determination of dihydroorotate dehydrogenase in Plasmodium falciparum was developed. The assay was applied to the evaluation of the effects of several antimalarial drugs on the enzyme. Treatment of both the asexual and gametocyte stages of P. falciparum in culture with menoctone, primaquine or the primaquine derivative WR 238605 led to depression of the enzyme activity, although the drugs did not appear to inhibit the enzyme directly.


Evaluation of the chromatographic procedure for the preparation of a high-purity C1-esterase inhibitor concentrate from cryosupernatant plasma. This report is an analysis of the parameters involved in the preparation of a C1-esterase inhibitor concentrate. We investigated ways to improve the purification by finding the optimal pH, ionic strength, and quantity of chromatographic media to use at each step. The optimisation of the process gave a greater than two-fold increase in both yield and purity over published methods. The product retained more than 50% activity upon dry heat-treatment at 80 degrees C for 72 h, and the yield of the heat-treated product was 180 I.U./kg of plasma.


Urinary testosterone measurement by gas chromatography after solid-phase extraction and high-performance liquid chromatography. A method was developed for the rapid determination of testosterone in urine. The procedure consists of solid-phase extraction (SPE) followed by high-performance liquid chromatographic (HPLC) clean-up before gas chromatographic determination. Recovery was evaluated by adding [3H]testosterone (10(4) cpm) to urine samples; the mean recovery of radioactivity after SPE and HPLC was 82%. Precision was estimated by repeated measurement of testosterone in four different urine samples; the coefficient of variation was 7.9% (95% confidence limits 6.1-11.4%). Accuracy was evaluated by standard addition and dilution assays; a linear relationship was found between the expected and observed values (r2 = 0.982). The method is rapid, effective and suitable for routine analysis.


Rapid purification yielding highly active 17 beta-hydroxysteroid dehydrogenase: application of hydrophobic interaction and affinity fast protein liquid chromatography. Homogeneous human placental 17 beta-hydroxysteroid dehydrogenase was obtained by a procedure consisting of two fast protein liquid chromatographic (FPLC) steps using Phenyl-Sepharose hydrophobic interaction and Blue-Sepharose affinity columns. In the first chromatography, the enzyme eluted only when an additional decrease in ionic strength was inserted after the ammonium sulphate concentration had reached zero, thus enhancing the separation. In the affinity chromatography, separation of contaminating proteins occurred at different stages of loading and washing. The specific elution of the enzyme by the co-factor NADP+ is very efficient in obtaining a homogeneous preparation in high yield. The rapidity of FPLC was further increased by a maximum simplification of the intermediate steps, and the whole procedure lasted only two days. This preparation has a yield of more than 50% and a high specific activity, catalysing the formation of 7.9 mumol of estrone from estradiol per minute at pH 9.2 and 23 degrees C. It has an apparent molecular mass of 35,000. This provides an efficient candidate for the purification of other membrane-associated proteins.


Separation of serum bilirubin species by micellar electrokinetic chromatography with direct sample injection. Four major bilirubin species in serum were separated by micellar electrokinetic chromatography with 25 mM sodium dodecyl sulfate (SDS) and 20 mM sodium tetraborate-boric acid buffer at pH 8.5. Due to the solubilization of the serum proteins by the SDS micelles, serum samples were injected directly into a 50 cm x 75 microns I.D. fused-silica capillary and complete separation of the four bilirubin species was accomplished within ca. 10 min without extensive sample pretreatment. Detection was performed by absorbance at 450 nm and average limit of detection was in the 6.0 microM concentration range. The usefulness of this method was demonstrated for the separation and detection of a number of bilirubin species present in pathological human serum samples.


Polypeptide marker and disease patterns found while mapping proteins in ascitis. To assess the protein composition of ascitis, 28 samples of ascitic fluid were obtained from patients admitted to Geneva University Hospital. The samples were analysed randomly and blindly by high-resolution two-dimensional polyacrylamide gel electrophoresis. The final visual evaluation was compared with the discharged summary and diagnosis. The protein pattern of ascitis was, as expected, very similar to normal or diseased plasma, with the exception of two spots which were present in ascitic fluids but not in the 200 plasma samples analyzed in parallel. After microsequencing, they proved to be beta-fibrinogen fragments. Several diseases showed distinct patterns, especially acute pancreatitis. A group of intense spots with an apparent molecular mass between those of alpha 1-antitrypsin and beta-haptoglobin were found in all ascitic fluid from pancreatitis cases (six patients). These spots had isoelectric points similar to those of alpha 1-antitrypsin and beta-haptoglobin and microsequencing revealed that they were three different fragments of alpha 1-antitrypsin.


Determination and identification of amiloride in human urine by high-performance liquid chromatography and gas chromatography-mass spectrometry. A simple and sensitive high-performance liquid chromatographic method was developed to screen and determine amiloride (I) in human urine. The detection limit of the method is 0.12 micrograms/ml and the recovery of amiloride from urine was 80.4-85.5% at different concentrations. The coefficients of variation were less than 2.8 and 4.4% for intra- and inter-assays, respectively. Total urinary excretion of I in 24 h after oral administration of 5 mg or 15 mg of I ranged from 22.0 to 33.3% of the total dose for three different subjects. I could be detected in urine up to at least 44 h after a 5-mg dose and 72 h after a 15-mg dose. A gas chromatographic-mass spectrometric (GC-MS) confirmatory method was established based on the methanolysis of I to methyl 3,5-diamino-6-chloropyrazine-carboxylate (II). The di-N-trimethylsilyl derivative of II showed very good GC-MS properties and provided reliable structure information for confirmation analysis of I. This is the first time that a reliable GC-MS method has been reported for the detection of urinary I.


Supercritical fluid extraction of polychlorinated biphenyls and pesticides from soil. Comparison with other extraction methods. A comparison is made of supercritical fluid extraction (SFE) with two other techniques widely used for the extraction of polychlorinated biphenyls (PCBs) and organochlorine pesticides in soil. Extraction conditions for the SFE of PCBs and pesticides were first determined. An experimental approach was set up to determine the influence of different extraction parameters such as pressure, extraction time, static and dynamic extraction, restrictor type and collection solvent for off-line SFE. The use of carbon dioxide at 50 degrees C and 20 MPa, 10 min static followed by 20 min dynamic extraction with collection in iso-octane were been found to be the optimum conditions. Two types of soil, with a low and high content of organic carbon, respectively, spiked with 16 PCBs and organochlorine pesticides with a wide range of volatility and polarity at a level of 5 ng/g dry matter, were used as test materials. Conventional solvent extraction gives a good extraction yield for soil with a low content of organic carbon, but for peat soil the recoveries decrease dramatically to 30% for DDE, DDT and PCB 138 and 153. The recoveries with Soxhlet extraction are good, but an extra clean-up step before analysis is necessary. SFE gives good extraction yields for PCBs and organochlorine pesticides, varying between 85 and 105% with a reproducibility of 5% for each component for both types of soil. SFE is a fast, clean and reproducible method for the extraction of PCBs and organochlorine pesticides from these two soil matrices.


Determination of organophosphorus pesticides in fruits by on-line size-exclusion chromatography-liquid chromatography-gas chromatography-flame photometric detection. The determination of organophosphorus pesticides in fruits by size-exclusion chromatography (SEC) on a polystyrene column coupled on-line to a gas chromatography (GC) system was unsatisfactory as a result of interfering peaks in GC. A liquid chromatography step on silica gel was therefore inserted between the SEC and GC steps to filter out polar by-products. Samples of fruit (apples, grapes and kiwi fruits) were extracted, then the extract filtered or centrifuged and injected into an automated on-line SEC-liquid chromatography-GC-flame photometric detection. Recoveries were about 95% and the detection limits about 1 ng/g.


Isolation, purification and partial characterization of tropomyosin and troponin subunits from the lobster tail muscle. In a search for an invertebrate muscle from which the muscle regulatory proteins could be obtained in a great quantity and at high homogeneity, the regulatory proteins, tropomyosin (Tm) and three subunits of troponin (Tn), have been isolated from the lobster tail muscle, purified and partially characterized. The calcium-sensitive ATPase of lobster myofibril was restored when purified lobster Tm and lobster Tn were added to actin. Quantitative SDS-polyacrylamide gel electrophoresis showed that the lobster muscle contains actin, Tm, Tn with a molar ratio 7:1:1 and that lobster Tn consists of three subunits, one of each I, C and T. Each subunit was identified according to its effect on the acto-S1 ATPase rate. The isomer composition in each fraction of purified Tn subunit and in Tm are different from the rabbit skeletal muscle proteins; Tm consists of a single species of polypeptide of M(r) 38,000; the TnT fraction appears to be homogeneous with M(r) 43,000; the TnI fraction contains five isomers, all showing similar isoelectric pH, differing in M(r) in the range from 28,000 to 31,000; two TnC fractions contain three isomers in total with a range of M(r) from 18,500 to 19,000. Further study of the lobster Tm elucidated that digestion by carboxypeptidase A gave rise to a homogeneous preparation of truncated and non-polymerizable Tm which is devoid of 11 residues at the C-terminus of the molecule. The C-terminal amino acid sequence of 11 residues is homologous to the thoracic isomer generated from Drosophila melanogaster Tm-I gene. The present study indicated that, despite heterogeneities owing to the occurrence of isomers, the lobster regulatory proteins serve as an invertebrate source of the proteins for structural and biophysical studies, alternative to vertebrate counterparts.


Unloaded shortening after a quick release of a contracting, single fibre from crayfish slow muscle. The time course of shortening at zero load was studied by the slack test method during tetanic contractions in isolated, single, slow muscle fibres of the crayfish. In 28 of 32 shortenings (from 14 different fibres) a biphasic shortening was seen, which consisted of an initial high-velocity phase lasting 3.3-20.8 ms and a following slow-velocity phase lasting for the entire time examined (up to 89.2 ms). Provided that the shortening occurred uniformly along the fibre length, velocity in the initial phase, V1, of the biphasic shortening was 14.4 +/- 3.4 (mean +/- SD, n = 10) microns s-1 per half sarcomere at Lo, the slack length, at 20 degrees C, while that in the second phase, V2, was 7.4 +/- 1.4 microns s-1 per half sarcomere. Lowering temperature decreased both V1 and V2 with Q10 = 1.4 for V1 and 2.0 for V2. Lowering the external Ca concentration from 15 mM, the standard, to 2 mM resulted in a tetanic tension below one-third of that at 15 mM Ca and decreased both V1 (t test; p < 0.01) and V2 (p < 0.001). Prestretching the fibre to 1.5 Lo had no significant effect on V2 (p < 0.3) but increased V1 (p < 0.001). The distance shortened during the initial high-velocity phase, LV1, was 4.0 +/- 1.8% Lo (mean +/- SD, n = 10) at 20 degrees C or about 0.14 microns per half sarcomere on average. LV1 was independent of the tetanic tension level when it was changed by lowering the external Ca concentration or temperature in the same fibre. Prestretching the fibre to 1.5 Lo, at which the sum of the active and the resting tension was lower than Po at Lo in two of three fibres, increased LV1 significantly (p < 0.001). The independency of LV1 from the tension level indicates that the initial high-velocity phase was not from shortening of some inert components in the fibre. One possibility is that the initial high-velocity phase was brought about by an acceleration of shortening by a compressive force, the origin of which has been discussed. The slow-velocity phase seemed to result from the crossbridge turnover with little exogeneous stress on myofilaments. Four different fibres exhibited an unloaded shortening with a constant velocity during the entire time examined (29.9-61.8 ms). This type of shortening had a velocity between the usual V1 and V2 values, suggesting that a compressive force accelerated the shortening during the entire time.


Relaxation rate of intact striated muscle fibres after flash photolysis of a caged calcium chelator (diazo-2). Single twitch fibres from lumbrical muscles of Xenopus have been loaded with the photolysable calcium-chelator diazo-2 by incubation in Ringer solution containing the membrane permeable acetoxymethyl ester (AM) form of diazo-2. Incubation caused a progressive slowing of tetanus rise and relaxation which is ascribed to calcium-buffering by unphotolyzed diazo-2 (Kd = 2.2 microM). After incubation, exposure to a brief UV flash caused a three to four fold increase in the rate of tension fall. A flash given 16-18 ms after the last tetanic stimulus (at 22-24 degrees C) resulted in 10% increase in relaxation rate compared with the control before incubation. A much bigger effect was observed when a flash was given half-way into the slow phase, where an 1.8-1.9-fold increase in relaxation rate, above the preincubation slope, was observed. It is concluded that rapid lowering of [Ca]i, and hence more rapid removal of Ca2+ from troponin, speeds up relaxation, indicating that calcium translocation is the major determinant of the rate of tension fall during the isometric phase of relaxation.


Sea urchin axonemal motion supported by fluorescent, ribose-modified analogues of ATP. The axonemal motion supported by fluorescent ribose-modified analogues, anthraniloyl ATP (Ant-ATP) and methylanthraniloyl ATP (Mant-ATP), was investigated. Ant-ATP and Mant-ATP supported good vigorous motion. A detailed study of the movement shows that the maximum beat frequencies (Vmax) were significantly lower with the analogues. However, Michaelis constants (Km) for beat frequency were also significantly lower than with ATP. Thus the net effect of changes in these two parameters, Vmax/Km, was similar for ATP and Ant-ATP and higher with Mant-ATP. Thus these fluorescent analogues are good substrates for axonemal movement. The consistently higher value of Vmax/Km, a measure of substrate selectivity, with Mant-ATP over Ant-ATP suggests a feature of the ribose binding site. Other significant differences in the movement with the fluorescent analogues are quantified in terms of kinetic measures of sliding velocity and bend propagation velocity.


Distribution of developmental myosin isoforms in isolated A-segments. Immunogold labelling was used to determine the distribution of myosin isoforms within the A-bands of developing chicken pectoralis muscles. Previous localization studies led to the suggestion that neonatal myosin is preferentially located in the centre of heterogeneous thick filaments that contain either embryonic or adult myosin in addition to neonatal myosin. To further explore the possibility that neonatal myosin may serve to nucleate thick filament assembly, a method was developed to isolate A-segments (arrays of myosin filaments) from myofibrils in the presence of MgATP. A-bands usually dissociate into thick and thin filaments in a relaxing buffer, but the inclusion of an antibody against M-line protein prevented separation of the thick filament array. Well-ordered A-segments, approximately 1.5 microns in length, were prepared from muscles 12, 29, 40 days, and approximately 1 year after hatching. After reaction with monoclonal antibodies specific for neonatal and adult myosins, the A-segments were labelled with gold-conjugated secondary antibodies prior to negative staining. An antibody which cross-reacts with embryonic myosin was used to localize that epitope in A-bands of myofibrils from day 1 and day 3 posthatch muscles. At ages where expression of neonatal myosin was high, extensive gold labelling of A-segments was observed in the electron microscope. However, no preferential distribution of antibodies was observed at any age, independent of whether embryonic or adult myosin was coexpressed with the neonatal myosin, suggesting that neonatal myosin is not segregated to any particular region in the A-bands of developing muscles.


Differences in maximal activation properties of skinned short- and long-sarcomere muscle fibres from the claw of the freshwater crustacean Cherax destructor. Single fibres of different sarcomere length at rest have been isolated from the claw muscle of the yabby (Cherax destructor), a decapod crustacean. Fibres of either long (SL > 6 microns) or short (SL < 4 microns) sarcomere length have been mechanically skinned and were maximally activated by Ca2+ and Sr2+ under various experimental conditions (ionic strength, in the presence of 2,3 butanedione monoxime (BDM)) to determine differences in their contractile properties. Isometric force was measured simultaneously with either myofibrillar MgATPase or fibre stiffness in both fibre types. The ultrastructure of individual long- and short-sarcomere fibres was also determined by electron microscopy. The long-sarcomere fibres developed greater tension (30.48 +/- 1.72 N cm-2) when maximally activated by Ca2+ compared with the short-sarcomere fibres (18.60 +/- 0.80 N cm-2). The difference in the maximum Ca(2+)-activated force can be explained by the difference in the amount of filament overlap between the two fibre types. The maximum Ca(2+)-activated myofibrillar MgATPase rate in the short-sarcomere fibres (1.60 +/- 0.27 mmol ATP l-1s-1) was higher, but not significantly different from the ATPase rate in fibres with long-sarcomeres (1.09 +/- 0.14 mmol ATP l-1s-1). As the concentration of myosin is estimated to be higher only by a factor of 1.22 in the short-sarcomere preparations there is no evidence to suggest that the myofibrillar MgATPase activity is different in the long- and short-sarcomere preparations. The maximum Ca(2+)-activated force (P0) of both short- and long-sarcomere fibres was quite insensitive to BDM compared with vertebrate muscle. Force decreased to 60.2 +/- 5.3% and 76.1 +/- 2.7% in the short- and long-sarcomere fibres respectively in the presence of 100 mmol l-1 BDM. The difference in the force depression between the long- and short-sarcomere fibres is statistically significant (p < 0.05). Fibre stiffness during maximum Ca(2+)-activation expressed as percentage maximum force per nm per half sarcomere was higher by a factor of 3.5 in short-sarcomere fibres than in long-sarcomere fibres suggesting that the compliance of the filaments in the long-sarcomere fibres is considerably higher than in the short-sarcomere fibres. Sr2+ could not activate the contractile apparatus to the same level as that seen by Ca2+ in either fibre type: the maximum Sr(2+)-activated force was (20 +/- 3%) and (63 +/- 3%) of the maximum Ca(2+)-activated force response in short- and long-sarcomere fibres, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)


Time course of rise of muscle stiffness at onset of contraction induced by photorelease of ATP. Isometric contraction (9-10 degrees C) of skinned fibres from rat psoas muscle was elicited by photorelease of ATP (1.3-1.4 mM), and force (F), in-phase (I) and quadrature (Q) stiffness were monitored. The sinusoidal length change for the stiffness measurement was approximately 0.09% fibre length and 200-1000 Hz. On photolysis, F and I initially fell, and then rose. The half-time of the rise of I was shorter than that of F. Contrary to the previous assumption, Q rose even earlier than I. Although the time courses of the rise of F and I were greatly affected by the presence of phosphate ion (Pi; 4 mM), its effect on Q was modest. We propose the following hypothesis: the rigor crossbridges, after binding ATP, initially enter a state characterized by high Q with low I and none or little F, and then, releasing Pi during passing the state with the increased I, reach the final force generating state.


A survey of local anesthetic usage in pediatric patients by Florida dentists. Local anesthetic toxicity in the child dental patient is a concern of the dental profession. A survey of 117 Florida dentists was conducted to determine their local anesthetic practices in treating children. Results indicated that a fifth of the respondents might use local anesthetic levels that could cause potential CNS toxicity in the smaller and younger children they treat. It is the responsibility of the dental profession to recognize the differences in treating this group of patients, and the serious consequences that might occur when maximum recommended levels of local anesthetic are exceeded.


The oral effects of orotracheal intubation in prematurely born preschoolers. This cross-sectional study of fifty-two prematurely-born children, ages two to five years, evaluated the long-term effects of oral intubation at birth on palatal architecture, crossbite malocclusion, and enamel structure. Seventy percent showed a high palatal vault, with palatal grooving in 25 percent; 36 percent had enamel defects in the maxillary primary incisors; and 17 percent had posterior crossbites. The mean palatal depth was 12.9 mm, as compared to 11.4 mm for a group of 45 nonintubated, normal healthy children (the difference was significant at alpha = 0.05). There are significant iatrogenic defects from prolonged neonatal intubation with this sample of preschoolers.


Management of aspiration and swallowing incidents: a review of the literature and report of case. This article has reviewed aspiration and swallowing incidents in a dental environment with particular emphasis on the young child and the medically, physically and mentally handicapped patient. A thorough review of prevention, signs and symptoms, management and documentation of these incidents has been presented. This review was presented to reaffirm the potential of these incidents and strives to educate the practitioner who treats these special populations. The case report applies reported guidelines for noting signs and symptoms, management and documentation. This case report also affirms the need to check all dental instruments before their use, as a safeguard against possible failure. Current policy for infection control maintains strict guidelines for asepsis. With the increasing use of steam autoclaving and cleansing products, an increased rate of failure may be seen in some instruments. Manufacturers should be contacted for instrument care and sterilization recommendations, as well as projected longevity of the instrument, if their sterilization procedures are followed. The economic implications of replacing instruments because of the protocol for maintaining asepsis will undoubtedly contribute to the rising cost of care. Prevention has been the goal of dentistry for decades and dental emergencies are not excluded. Preparation for such incidents will prevent many incidents and allow for proper patient management should they occur.


Expression of CA 50 in normal human skin: comparative study of CEA and CA 19-9. Normal skin specimens from different body sites were estimated for the expression of CA 50 in comparison with CA 19-9 and CEA. Expression of CA 50 and CA 19-9 was observed in the ductal system and a small part of the secretory portion of eccrine units, but never in any other skin components including apocrine units. CEA was expressed on the ductal and a small part of the secretory portion of apocrine units as well as on those of eccrine units. However, the expression pattern and intensity in eccrine units were different from those of CA 50 or CA 19-9. We concluded that CA 50 was strongly expressed in the normal ductal system of eccrine units and hence could be standardized to stain the eccrine ductal portion and that immunohistochemical comparative studies with CA 19-9 or CEA might be a good methodological technique for discriminating an eccrine or apocrine origin for some skin disorders.


Prevalence of hidden caries.

Prevalence of hidden caries. Clinical evaluations of the first and second permanent molars were compared with radiographic evaluations of the same teeth, for 359 patients (between six and eighteen years old) of the pediatric department of the Academic Centre for Dentistry Amsterdam (ACTA). The teeth were professionally cleaned and dried before the clinical evaluation. Despite the thorough examination, taking into account the fact that all discolorations and/or decalcifications were noted, 15 percent of the teeth that did not show signs of enamel caries clinically, showed a dentine lesion on the bitewing radiograph. These 15 percent of teeth with hidden caries were found in 7.5 percent of the population studied. The absence of clinical signs of occlusal enamel caries or dentine caries does not guarantee a sound dentition. Sealant treatment of teeth that are sound (both clinically and radiographically) is justified, therefore, soon after tooth eruption.


Demographic and psychosocial characteristics of western Pennsylvania school-age tobacco users. A 25-item tobacco usage questionnaire was presented to 2,189 subjects in grades 1 through 12 in a school district in Western Pennsylvania. It was found that thirty percent (30 percent) of the subjects were former or current cigarette smokers. Approximately 20 percent of the subjects reported using snuff, while approximately 16 percent reported using chew. Use of smokeless tobacco was found to be associated with such variables as grade level, school grade-average, parents' education, parents' use of tobacco, peer tobacco usage, and participation in certain sports.


Growth and collagen synthesis of cultured neurofibroma fibroblasts. Cells from cutaneous neurofibromas of three patients with von Recklinghausen's disease and skin fibroblasts from four healthy adults were cultured. After two passages, DNA and collagen syntheses were determined by measuring incorporation of [3H] thymidine and [3H] proline respectively, and expressed as the values per unit DNA content. These values from neurofibroma cells were increased by 54% in DNA synthesis and 60% (p < 0.05) in nondialyzable hydroxyproline synthesis, suggesting that neurofibroma cells in culture even after several passages still possess those characteristics corresponding to their pathological features in vivo. Collagen synthesized by neurofibroma cells appears to be normal as judged by intracellular degradation rates and SDS-polyacrylamide gel electrophoresis.


Effects of malnutrition on development of experimental pressure sores. Using food-deprived rabbits we investigated the effects of nutritional deficiency on the development of pressure sores. The body weight of these animals was decreased significantly from normal. Organ weights of liver, heart, spleen, kidney, and testis were significantly decreased from normal as well. Protein deficiency in these animals was indicated by serum tests, as well as by histologic features of liver and testis and ultrastructural findings on fibroblasts. We produced lesions in malnourished as well as normal rabbits by exposing their skin to a balloon-produced compressive force of 120 +/- 10 mmHg for 4 hours. Biopsies were taken 1, 2, and 3 days after the pressure application. Histologic findings at each time were as follows: At day one, the degree of ischemic skin destruction in the malnourished animals was more severe than that in the normal ones, and thrombi were occasionally seen in the malnourished cases. At day two, proliferation of fibroblasts and macrophage infiltration were evidenced in the normal animals, whereas signs of collagen fiber degeneration as well as microthrombi were seen in the malnourished ones. At three days, epidermal cells covered the lesions in the case of normal animals; however, massive necrosis of the epidermis was still recognized in the malnourished rabbits. Thus, the healing process of pressure sores was strongly suppressed in the malnourished animals. This suppression was attributed to the reduction in fibroblast proliferation, capillary formation, macrophage infiltration, and also to the low level of epidermal cell proliferation.


Children are expensive.

Children are expensive. Rearing children is not cheap. Family and socioeconomic costs are explored.


A case of subepidermal blister disease associated with autoantibody against 450 kD protein. We report a case of subepidermal blister disease with eruptions distributed on the trunk and extremities. The blisters regressed after administration of corticosteroid and cyclophosphamide. An immunoblot analysis revealed that the patient's serum reacted exclusively with a 450-kD epidermal polypeptide. It did not react with a 230-kD or a 180-kD bullous pemphigoid antigen. The immunofluorescence study showed the patient's serum reacted with the basement membrane zone of human skin.


Tuberculosis: revisited.

Tuberculosis: revisited. Recent upsurge of tuberculosis (TB) has been reviewed. Clinical features of TB have been briefly described along with possible risk of transmission. Methods for prevention of transmission have been reported, including suggestions pertaining to prevention in the dental office.


Localized basaloid follicular hamartoma. We report a 39-year-old Japanese woman with a 30 x 10 mm, skin-colored, soft, hairless, irregularly elevated plaque in the left retroauricular area. Histologically, it consistently showed features of atypical hair follicles; some showed immaturely developed basaloid epithelial islands invaginated with condensed stromal cells, and others were replaced or associated with solid strands, branching cords, or lace-like networks of undifferentiated basaloid cells. Although the clinical appearance of a typical localized basaloid follicular hamartoma is a plaque of alopecia, this case seems to be a variant, because the histological findings are so characteristic of the entity. The relationship of basaloid follicular hamartoma to other hair-follicle derived tumors is also considered.


Gender differences in the characteristics of dental services provided for children. The purpose of this project was to describe the differences and similarities in the practice of dentistry for children by male and female dentists. A once-mailed survey with a series of questions regarding the characteristics of their dental practices was sent to a random sample of 3,000 dentists. There were 1,154 useable responses (39 percent). Approximately 8 percent of the respondents were women, which is representative of the percentage of female dentists practicing in the United States. Chi square analysis was employed. There were differences in the practice characteristics of the male and female dentists responding to this survey. The female respondents were: significantly younger; had been in practice for a shorter period of time; more likely to be employees rather than solo practitioners; more likely to treat younger patients; less likely to do comprehensive orthodontic treatment and more likely to use physical restraints than their male counterparts.


BCC-associated amyloidosis with a peculiar pattern of deposition. An 83-year-old Japanese woman with lepromatous leprosy had been treated in a leprosarium. More than 10 years ago, she developed a dome-like brown tumor on the dorsum of the nose which showed the histology of basal cell carcinoma. The lesion was a well bordered, rounded tumor with tumor cell nests on its periphery extending toward the center as cords of tumor nest and with amyloid depositions between the cords. The tumor nests and amyloid lessened toward the center of the tumor, being replaced by collagen fibers. Amyloidosis showing this peculiar pattern of transition was reported with a review of published cases.


A rare association of localized gigantism with tuberous sclerosis. An unusual association of localized gigantism with hypertrophy of the long bones and soft tissues in the left lower limb in an 18-year-old male with tuberous sclerosis (TS) is reported. The significance of this association is discussed from the point of view of its common neural crest origin during embryogenesis.


Subcorneal pustular dermatosis and IgA multiple myeloma. A case of subcorneal pustular dermatosis associated with IgA multiple myeloma is described. The significance of the combination of the two diseases is not yet known, but the association is certainly more than coincidence.


Autoimmune progesterone dermatitis.

Autoimmune progesterone dermatitis. A 35-year-old woman had recurrent urticarial erythemas on her trunk and extremities for 3 years. The eruptions appeared regularly 3 to 5 days prior to menstruation and persisted for several days. The patient showed a positive skin test response to progesterone and had circulating antiprogesterone IgG antibodies. A speculative concept of the possible autoimmune damage to the ovary, the major organ producing progesterone, is discussed.


Tuberous sclerosis and spina bifida occulta. An interesting occurrence of tuberous sclerosis and spina bifida occulta is described, highlighting their unique and previously undescribed association in a 5-year-old boy.


Two patients with solar urticaria manifesting pruritic erythema. Two patients with solar urticaria are described. They complained of transient pruritic erythema, but not frank wheal, on sun-exposed areas. However, urticarial wheals were elicited by polychromatic radiation in the ultraviolet and visible light ranges; the action spectra for solar urticaria ranged from 400 to 550 nm in case 1 and from UVA to 550 nm in case 2. In patients manifesting only transient pruritic erythema on sun-exposed areas, phototests are essential for making a confident diagnosis of solar urticaria.


Analytic models for nonlinear curve-fitting of forward-rate binding data, with applications to hemoglobin. The experimental procedure of Perrella et al. (1983, J. Biol. Chem. 258, 4511-4517) can reveal most of the intermediate ligation states of hemoglobin either at equilibrium or in a kinetic reaction. In this paper, a model of stepwise ligand binding, under conditions of negligible reverse reaction, is derived with particular attention to hemoglobin. The model leads to a two-phase procedure. First, the ratios of the forward rates are determined by fitting the model to normalized concentrations vs. saturation. Second (optionally), an auxiliary relation is used to deduce the scale factor for the rates that make the model consistent with time information if such is available. The procedure has been applied to hemoglobin binding of the ligands carbon monoxide (this paper) and ferricyanide (manuscript in preparation).


Characterization of bacterial growth on solid medium with image analysis. Alkaliphilic bacterial strains producing the enzyme cyclodextrin glucanotransferase were cultivated on solid agar medium containing an indicator system detecting the enzyme. The growth of the colony and the surrounding diffusion zone, due to the enzyme, were measured by the image analysis during the cultivation. It was possible to differentiate between relatively similar clones by observing quantitatively the changes at and around the colony. Optimal experimental conditions for such measurements are discussed. The image analysis technique provides a potential tool for characterizing microbes grown on solid media.


Cell growth on immobilized cell-growth factor; 4: Interaction of fibroblast cells with insulin immobilized on poly(methyl methacrylate) membrane. Insulin was immobilized on the surface-hydrolyzed poly(methyl methacrylate) membrane and the growth acceleration of mouse fibroblast cells, STO, by the immobilized insulin was investigated. It was found that insulin remains immobilized on the surface of nonbiodegradable membrane and interacts specifically with receptors existing on the biological membrane of fibroblast cells. The growth acceleration by immobilized insulin was enhanced by introduction of a spacer arm between insulin and the immobilization matrix. The amount of receptor proteins present on the biological membrane of fibroblast cells after culturing with insulin, immobilized on nonbiodegradable polymer membrane, was much higher than that after culturing with free insulin, implying the suppression of down-regulation in the case of immobilized insulin.


Bioluminescence analysis of NAD(P)H and NAD(P)+ preventing mutual interference by selective nucleotide destruction and enhanced specific light emission. Improved bioluminescence analysis of pyridine nucleotides has been designed based on the fact that the luminescence intensity expresses the velocity of the light formation. The bacterial luciferase system is, in principle, composed of two reactions with two different velocities, one for energy supply by the oxidation of NAD(P)H and the other for the subsequent light generation. The rate setting can be arranged such that an emission maximum is produced 30 to 40 s after mixing the sample with the light-yielding solution, hence providing for a convenient analytical performance. The maximal intensity which is easily recorded, e.g., by a tracking volt-meter, is proportional to the concentration of the reduced nucleotide. Discriminative analysis of the various pyridine nucleotides is facilitated by selective destruction of the oxidized forms with alkali and the reduced forms with acid. Erroneous conversion of NAD(P)H to NAD(P)+ may be induced by haemoglobin in a tissue sample but this is prevented by the presence of 2 mM ascorbic acid at the instant of the acidification. Simultaneous coupling of the ongoing reduction of a pyridine nucleotide to the oxidation in the bacterial luciferase system generates a light-yielding cycle which offers important advantages. With NAD(P)+ as the analytic target compound, direct measurement replaces a preceding separate conversion to NAD(P)H. The four nucleotide forms become determinable in a sample by combining selective destruction of either the reduced or oxidized species with a nucleotide-specific reduction in the cycle. Discriminative analyses are furthermore facilitated by the enhanced emission which is due to the energy derived from the continuous specific reduction, whereas initial light signals from side reactions fade out. It is often possible to suppress disturbing analytical errors by the design of the light-yielding cycle. If the rate of the dehydrogenase reactions is kept low compared with the overall rate of the luciferase system, moderately impaired function of some of its components may only give rise to a slight and tolerable decrease in emission intensity. Kinetic evaluations and model experiments are presented and supplemented with applications to tissue samples.


Formation of proline thiohydantoin with ammonium thiocyanate: progress towards a viable C-terminal amino-acid-sequencing procedure. Pure amino acid thiohydantoins are required as reference standards for development of C-terminal-sequencing procedures based on thiohydantoin formation of the C-terminal amino acids of peptides and proteins. Proline thiohydantoin was prepared using a straightforward method involving reaction of acetylproline with ammonium thiocyanate. It was characterized by UV spectrophotometry, mass spectrometry and back-hydrolysis to the free amino acid. These data establish unequivocally that the thiocyanate procedure is applicable to proline as well as to the other common amino acids. This work also validates earlier claims that proline thiohydantoin can be prepared by reaction with thiocyanic acid.


Determination of endothelin by an immobilized receptor assay utilizing a 96-well format. Bovine cerebellar membranes immobilized on 96-well microtiter plates provide receptors for 125I-labeled endothelin-1 as the basis for a competitive binding assay. Adsorption of the membranes to a surface does not significantly alter the ligand-receptor interaction and reduces non-specific binding to 3-7% of total binding compared to 10-20% for a filtration technique. Considerable savings in reagents are realized since assays can be performed in 100 microliter volumes with only 10-20 micrograms of membrane protein. The 96-well format allows the rapid quantitation of large numbers of samples, and the assay is especially attractive in that it utilizes readily available reagents and equipment without the need for specific antibodies. The endothelin-receptor-based assay may be used to measure conversion of big endothelin-1 to endothelin-1 in aqueous assays. Since the presence of serum does not affect this method, tissue culture medium may be directly analyzed for endothelin production by cultured cells. All three isoforms of endothelin are detected, and the specificity of the receptor is retained since fragments and precursor forms of endothelin are not recognized. In cases where multiple endothelin isoforms may be present or where specificity of binding is in question, this assay may be used in conjunction with high pressure liquid chromatography to distinguish active peptides.


A simple vacuum transfer device for nucleic acids. We present a new vacuum device intended for transferring nucleic acids from agarose gels onto hybridization membranes. The apparatus is drawn so that plastic sheets serve as substitutes for the elaborate, but cumbersome and unnecessary, locking systems mounted on all the commercial blotters. This substitution results in (1) a much more easy and rapid set-up of the transfer and (2) a very simple framework allowing the possibility of low-cost manufacturing.


A fluorimetric assay for the effects of cytolytic toxins on the transport properties of resealed erythrocyte ghosts. We prepared resealed erythrocyte ghosts loaded with SPQ and chloride. We demonstrated that these membranes were still functional, as they were capable of exchanging anions, most probably through the band-3 protein. When cytolytic toxins (Escherichia coli hemolysin and Staphylococcus aureus alpha-toxin) were offered to the resealed ghosts, the internal SPQ was released. This could be attributed to the formation of toxin-induced ion channels into the ghost membrane that were so large that SPQ could escape through them. This release was actually independent of the anion-exchanging protein, since DIDS had no inhibitory effect on it. Due to their simplicity, and because they do not lyse, erythrocyte ghosts may serve as useful models to study the action of cytolytic pore-forming toxins. To assess the validity of these model membranes we compared results obtained using RBC and resealed erythrocyte ghosts as targets for the toxin, finding complete consistency. Pre-assembled toxin channels could also be studied on the ghosts. Applying different proteolytic enzymes to the external compartment after channel formation, we found that performed E. coli hemolysin pores were at least partially destroyed by enzymatic digestion.


Statistical testing of equality of two break-points in experimental data. Two examples in quantitative biology are examined to emphasize the need for two-phase regression models: the osmotic behaviour of cells and the non-linear temperature kinetics of membrane-bound enzyme systems. Existing statistical techniques are inadequate to test the equality of break-points of two data sets for specific reasons. We suggest here a pragmatic solution by way of a computer programme useful in applying two-phase regression models to such data sets wherein a decision needs to be made whether the critical transition differs or not.


Characteristics of sympathetic reflexes evoked by electrical stimulation of phrenic nerve afferents. In chloralose-anaesthetized cats, sympathetic reflex responses were recorded in left cardiac and renal nerve during stimulation of afferent fibres in the ipsilateral phrenic nerve. In cardiac nerve, a late reflex potential with a mean onset latency of 75.6 +/- 13.8 ms was regularly recorded which, in 20% of the experiments, was preceded by an early, very small reflex component (latency between 35 and 52 ms). In contrast, in renal nerve only a single reflex component after a mean latency of 122.1 +/- 13.1 ms was observed. Bilateral microinjections of the GABA-agonist muscimol into the rostral ventrolateral medulla oblongata resulted in a nearly complete abolition of sympathetic background activity and in an 88% reduction of the late reflex amplitude with only small effects on the latency of the evoked potentials. Under this condition, an early reflex component was never observed to appear. After subsequent high cervical spinalization, the residual small potentials which persisted after bilateral muscimol injections were completely abolished and in cardiac nerve an early reflex potential with a mean latency of 45 +/- 10 ms was observed in all but one experiment. The early reflex was therefore referred to as a spinal reflex component which, however, is suppressed in most animals with an intact neuraxis. In the renal nerve a spinal response was only observed in one experiment after spinalization. The results suggest that sympathetic reflexes evoked by stimulation of phrenic nerve afferent fibres possess similar spinal and supraspinal pathways as previously described for somato-sympathetic and viscero-sympathetic reflexes.(ABSTRACT TRUNCATED AT 250 WORDS)


Involvement of dopamine in control of renal blood flow. Previous studies have shown that the renal nerve supply in the dog contains a population of dopaminergic vasodilator neurons. In order to obtain more information on the possible physiological functions of these neurons, we examined two situations in which there is existing evidence for active intrarenal vasodilation--the reflex response to coronary artery occlusion and the autoregulatory response to reduced renal perfusion pressure. We were unable to confirm published reports that coronary artery occlusion evokes renal vasodilation. However, our results support the view that dopaminergic nerves participate in maintenance of renal blood flow and glomerular filtration at perfusion pressures around the lower limit of autoregulation.


Additive effects of dopamine and 8-OH-DPAT microinjected into the nucleus ambiguus in eliciting vagal bradycardia in rats. The effects of combined microinjection into the nucleus ambiguus (NA) of dopamine (DA) and the 5-hydroxytryptamine-1A (5HT-1A) receptor agonist 8-hydroxy-2-[di-n-propylamino]tetralin (8-OH-DPAT) on arterial pressure (AP) and heart rate (HR) were studied in 24 urethane-anaesthetized, artificially ventilated spinal (C1) rats. Sites from which bradycardia was elicited by microinjection of L-glutamate (GLU) were selected for microinjection of DA and 8-OH-DPAT into the NA. Microinjections of 8-OH-DPAT (60-300 pmol in 10 nl) elicited a dose-dependent bradycardia; the HR responses elicited by microinjection of 300 pmol of 8-OH-DPAT were significantly larger (-34.0 +/- 3.0) than responses elicited by the threshold dose of 60 pmol (-3.1 +/- 0.1 bpm). The onset latency of the HR responses elicited by the 300 pmol of 8-OH-DPAT was 6.5 +/- 0.5 s and the peak was reached in 40.0 +/- 8.0 s. The duration of these responses was 615 +/- 3.5 s. Microinjection of DA (1 nmol in 10 nl) within 3-5 s from the time of 8-OH-DPAT microinjection (60 pmol in 2 nl) into the NA at sites previously shown to elicit decreases in HR following microinjection of GLU, produced significant additive effects in eliciting bradycardia (20.4 +/- 2.9 bpm) when compared with decreases in HR elicited by microinjection of DA (11.1 +/- 1.8 bpm) or of 8-OH-DPAT (3.1 +/- 0.1 bpm) alone. There were no changes in AP after microinjections of DA, 8-OH-DPAT or of these two substances combined.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of the rostral ventrolateral medulla in the generation of synchronized sympathetic rhythmicities in the rat. In artificially ventilated, paralyzed rats anesthetized with Nembutal or urethane, power density spectral analysis (PDS), using direct FFT algorithm, was used to quantify rhythmicities in the sympathetic cervical and lumbar nerves after bilateral microinjections into rostral ventrolateral medulla (RVLM) of CoCl2 and MgCl2--unspecific synaptic transmission blockers. Later overall sympathetic activity, phrenic nerve discharge, heart rate and arterial blood pressure were recorded. Block of synaptic transmission in RVLM was tested by elimination of sympathoinhibitory or sympathoexcitatory reflex responses to aortic nerve and vagal afferents stimulation respectively. In animals vagotomized with bilateral section of carotid sinus nerve the power in all frequency bands was very significantly reduced to a level not different from that which remained after spinal cord transsection. If carotid baroreceptors were intact, a small peak corresponding to cardiac frequency band persisted. Overall, non-synchronized sympathetic activity and arterial blood pressure increased. All effects were transient, lasted up to 15 min, and could be reproduced several times in one experiment. Respiratory rhythmic activity was unchanged yet respiratory-sympathetic synchronization was abolished. It is concluded that RVLM reticulospinal sympathoexcitatory neurons are responsible for non-synchronized tonic sympathetic activity but are not able to generate synchronized sympathetic rhythms. Synaptic input, presumably inhibitory, either from local neuronal circuits within ventral medulla and/or from other brain stem neuronal circuitries is needed to shape out the flexible pattern of sympathetic oscillations.


Different modes of dampening influence from baroreceptors are determined by the functional organization of the NTS neuronal network. Simultaneous recordings of several neurones of the first relay station of baroreceptor afferents show that its general activity-dampening influence is realized via the common brainstem system (CBS) which itself controls the processing on the neurones of the nucleus of the solitary tract (NTS). This feedback system maintains the degree of activity which is necessary for the ongoing behaviour as long as it fits to the environmental situation. The output of the NTS is determined partly by the CBS, partly by the properties of the peripheral afferent input, partly by the dynamic functional organization of the local circuits and partly by influences from other brain areas.


Current data and ideas on digestive sensitivity. Remarkable advances have recently been made in the study of visceral sensitivity. Both electrophysiological and histological data emphasize the richness and the complexity of information elicited in the visceral area including the digestive tract. In addition these afferents are largely involved in physiological mechanisms which are not only restricted to the visceral area. The current knowledge of visceral sensitivity leads us to reconsider the classical ideas and concepts previously accepted in this field and raises questions to help gain a better comprehension of sensory physiology in general.


Mesencephalic cuneiform nucleus and its ascending and descending projections serve stress-related cardiovascular responses in the rat. The aim of the present study was to explore the neuroanatomic network that underlies the cardiovascular responses of reticular formation origin in the region of the cuneiform nucleus (CNF). The study was performed in urethane anesthetized male Wistar rats. The left iliac artery was supplied with a catheter for the measurement of systemic blood pressure. Low intensity electrical stimulation of the mesencephalic reticular formation (MRF) in the vicinity of the CNF always resulted in pressor and bradycardiac responses, whereas stimulation in the parabrachial nucleus (PB) and Kölliker-Fuse nucleus (KF) led to a pressor response and a small tachycardiac response. The cuneiform area may be placed in the center of a circuit that serves a specific autonomic response pattern to stress: parallel activation of the sympathetic (pressor response) and parasympathetic limb (bradycardia). The efferent connections of the effective stimulation sites in the MRF and the CNF area, were investigated by anterograde tracing with the lectin Phaseolus vulgaris leucoagglutine (PHA-L). The CNF sends descending fibers to the gigantocellular reticular nuclei (GI), the motor nucleus of the vagus (DMNV) and nucleus tractus solitarius (NTS). These projections are probably involved in the bradycardiac response to stimulation. The descending pathway to the NTS/DMNV and GI may therefore be the parasympathetic limb of the circuit. Furthermore, the CNF sends ascending fibers to limbic forebrain areas and descending fibers to the PB-KF complex. The KF in its turn projects to the rostroventrolateral medullary nucleus (RVLM) and the intermediolateral cell column (IML). These latter projections are partly involved in producing the pressor response and thereby represent the sympathetic limb of the circuit. Accordingly, the transection of the descending fibers from the CNF to the PB-KF complex resulted in a decreased pressor and an increased bradycardiac response. This suggests that a baroreceptor reflex-induced bradycardia which results from blood pressure increase can be excluded as the origin of the stimulation-induced bradycardia, and that the pressor and bradycardiac responses are two independent moieties. It cannot be excluded that ascending fibers from the CNF are also involved in producing the pressor response. On the basis of the present physiological and neuroanatomical study, a brain circuit has been proposed in which the cuneiform nucleus has a central position. The described brain circuit may serve a passive coping strategy to novel, painful or threatening stimuli during which the animals show orientation/attention or freezing behavior accompanied by a bradycardiac and pressor response.


Multiple modes of operation of cardiac autonomic control: development of the ideas from Cannon and Brooks to the present. Since Cannon's time much emphasis has been placed on reciprocal control of the organ function by the autonomic nerves while other modes of control has been neglected. In our laboratory, prompted by Dr. Brooks, we initiated the study of the autonomic control of the heart by recording simultaneously the activity of cardiac sympathetic and parasympathetic nerves under a variety of conditions in chloralose anesthetized dogs. We have demonstrated that the central nervous system exercises reciprocal as well as non-reciprocal pattern of control over the two arms of the autonomic outflows under different physiological and behavioral conditions, and that each mode of control has an important functional significance.


Specialized functional pathways are the building blocks of the autonomic nervous system. The autonomic nervous system supplies each type of target organ via separate pathways which consist of sets of pre- and postganglionic neurones with distinct patterns of reflex activity. This has been firmly established for the lumbar sympathetic nervous system to skin, skeletal muscle and viscera, for the thoracic sympathetic outflow to the head and for several parasympathetic systems. In principle, that was already known by Langley. The specificity of the messages that these pathways transmit from the central nervous system arises from integration within precisely organized pathways in the neuraxis. The messages travel along discrete functional pathways and are transmitted to the target tissues via close neuroeffector junctions. Integration in the periphery occurs within each pathway, both in ganglia and at the level of the effector organs. We still need to understand how the central messages get through without distortion and how they control the diverse functions of the vasculature and viscera.


The meaning of H2O2 generation in carotid body cells for PO2 chemoreception. The rat carotid body is able to generate H2O2 in type-I cells with the aid of an electron-transferring chain with cytochrome b as the major component as it can be detected by spectrophotometry as well as confocal laser-microscopy. This cytochrome b is reducible by hypoxia, but not by cyanide, indicating that it does not participate in the energy production by the respiratory chain. The carotid body possesses a glutathione peroxidase (GPO) which scavenges H2O2 and other organic hydroperoxides. The nervous chemoreceptor discharge can be inhibited by external application of hydroperoxides with a similar half maximal value (60-80 microM) as used to stimulate GPO. A hypothetical signal chain is described which suggests the involvement of cytochrome b as an O2 sensor in PO2 chemoreception of the carotid body and the degradation of H2O2 by glutathione to control the K(+)-conductivity of carotid body cells.


Neuronal mechanisms responsible for ongoing activity of rabbit superior cervical ganglion neurons. The action potentials evoked by stimulation of single preganglionic nerve fibres or of single neurons of the ganglion were recorded from postganglionic nerves of rabbit superior cervical ganglion. The averaging technique was used to improve the nerve signal-to-noise ratio. It was found by dividing the area by the area that single preganglionic nerve fibre discharges, as an average, 15 neurons of the ganglion. Action potentials were also recorded from preganglionic nerves, either appearing synchronously with the intracellular spikes recorded during ongoing activity of the ganglion neuron, or being evoked by stimulation of a single preganglionic nerve fibre. It was found by dividing the area by the area that each ongoing spike in the ganglion neuron is preceded by firing, as an average, of three preganglionic nerve fibres. Thus, 45 ganglion neurons are expected to be discharged by a preganglionic volley during their ongoing activity. This number is much lower than that found in our previous experiments (about 100 neurons). The difference suggests that in the majority of the ganglion neurons the ongoing discharges need summation of excitatory effects produced by a few converging preganglionic nerve fibres (multiple input) rather than being evoked by a single preganglionic nerve fibre (single input). Methacine, a selective muscarinic antagonist (0.3-0.5 mg/kg, i.v.), decreased by about 10% the rate of the multiple input-induced ongoing spikes while not affecting the single input-induced ongoing spikes in the ganglion neurons. This effect is probably due to the increase in the duration of after-hyperpolarization observed after the application of methacine.(ABSTRACT TRUNCATED AT 250 WORDS)


Local cerebral blood flow responses in rats to hypercapnia and hypoxia in the rostral ventrolateral medulla and in the cortex. The effects of hypercapnia and hypoxia on two local cerebral blood flows in the parietal cortex (PC-BF) and rostral ventrolateral medulla (RVLM-BF) were examined using laser Doppler flowmetry in anesthetized rats. Hypercapnia for 45 s duration at the end-tidal CO2 between 5% and 10%, induced by increasing inspiratory CO2, increased both cerebral blood flows and systemic blood pressure in a degree-dependent manner. The response of RVLM-BF was significantly stronger than that of PC-BF. Both cerebral blood flow responses to hypercapnia were not influenced by cutting peripheral chemoreceptor afferent nerves. Hypoxia for 45 s duration at the end-tidal O2 between 12% and 6%, induced by decreasing inspiratory O2, produced an increase of similar magnitude in both RVLM and PC local blood flows in a degree-dependent manner and a decrease in systemic blood pressure. The responses of both PC-BF and RVLM-BF to hypoxia were significantly diminished after cutting peripheral chemoreceptor afferent nerves. It is concluded that: (1) the RVLM-BF is much more sensitive to hypercapnia than the PC-BF; and (2) activation of peripheral arterial chemoreceptors possibly contributes to hypoxia-induced increase in the RVLM-BF and PC-BF.


Electrophysiological study on the vagal innervation of the adrenal gland in the rat. Action potentials evoked by stimulation of the ventral or dorsal subdiaphragmatic vagal nerve trunks were recorded from the adrenal branch of the splanchnic nerve in the urethane-anesthetized rat. Action potentials were clearly demonstrated after averaging over 50 times by a computer. In some experiments action potentials evoked by stimulation of the adrenal branch of the splanchnic nerve were observed in the celiac branch of the vagus nerve. The observations indicate the existence of a nervous pathway from the ventral and dorsal subdiaphragmatic vagus nerve to the adrenal gland, and the conduction velocities (0.32-0.91 m/s) suggest that the majority of the nerve fibers belong to the non-myelinated C-fiber group.


Delayed puberty in uremia: pituitary-gonadal function during short-term pulsatile luteinizing hormone-releasing hormone administration. Pubertal development is frequently delayed or disordered in children with chronic renal failure. Both neuroendocrine and peripheral alterations due to uremia have been hypothesized to explain the impairment in the pituitary gonadal axis. The aim of the present study was to evaluate quantitative (immunological) and qualitative (biological) LH secretion, as well as FSH and sex steroids, before and during 7 days of sc LHRH administration (136-150 ng/kg bw every 120 min) in 5 uremic children (13.1-14.8 yr) with delayed puberty. Six nonuremic children (13.2-17.8 yr) with delayed puberty underwent the same schedule and served as control group. On day 0 mean immunoreactive LH (I-LH) levels were higher in uremic (4.5 +/- 0.9 mIU/ml) than in nonuremic (1.9 +/- 03 mIU/ml; p < 0.05) subjects while no differences were observed in bioactive LH (B-LH) levels (2.9 +/- 0.7 mIU/ml vs 2.4 +/- 0.3 mIU/ml). In both groups of subjects testosterone was at prepubertal levels. Spontaneous I-LH and B-LH pulses were observed sporadically in both uremic and nonuremic subjects. Short-term pulsatile LHRH administration induced significant increases in B-LH, I-LH, FSH and testosterone. The B/I LH ratio increased from day 0 (0.7 +/- 0.2) to day 7 (1.3 +/- 0.4; p < 0.05) in uremics while it showed wide fluctuations in nonuremic subjects. On day 7, 4 uremic and 5 nonuremic subjects showed a pulsatile release of B-LH after exogenous LHRH pulses. Our data document that in uremia there are qualitative as well as quantitative abnormalities in pituitary gonadal secretion.(ABSTRACT TRUNCATED AT 250 WORDS)


Immunohistochemical detection of estrogen and progesterone receptors in the normal urinary bladder and in pseudomembranous trigonitis. According to recent studies showing the presence of estrogens receptors (ERs) in the human female lower urinary tract, we performed ER and progesterone receptor (PR) assays in fresh frozen and paraffin embedded biopsies taken from the urinary bladder. Fourteen females undergoing endoscopy during staging for gynecological cancer (endometrium, cervix, ovary) and 15 women complaining of recurrent abacterial cystitis (pseudomembranous trigonitis) were enrolled in the study as Group 1 and Group 2, respectively. After informed consent, they were submitted to cystoscopy, during which two biopsies were taken: one on the trigonum and the other on the bladder lateral wall. ERs were identified in both groups only in the trigonum (7/14 patients in Group 1 and 8/15 in Group 2), whereas the bladder lateral wall always stained negative. PRs were found at both sites in both groups (11/14 cases on the trigonum and 7/14 on the bladder lateral wall in Group 1; 11/15 and 2/15 respectively in Group 2). Morphological localization of PRs showed intense omogeneous staining in the nuclei of the stromal fibroblasts too. A clear correspondence between the presence of steroid receptors at the squamous metaplasia of the trigonum was observed. These data are discussed speculating about a possible endocrine pathogenesis of pseudomembranous trigonitis.


Effects of the administration of thyroid hormone on the plasma levels of atrial natriuretic peptides and on atrial myoendocrine cells in the rat: an immunochemical, ultrastructural, and stereological study. The effects of an altered thyroidal status on the levels of immunoreactive (ir-) atrial natriuretic peptide (ANP) in serum and in the right and left atria, as well as on the subcellular structures of atrial myoendocrine cells were explored in groups of male Sprague Dawley rats given the vehicle or triiodothyronine in the toxicological dose-range (50 micrograms/100 g bw/day) for 0,5, 1,2,4,7 or 14 days. Plasma levels of ir-ANP were 30% higher in T3-treated rats compared with controls at 0,5 and 1 day after hormone administration and then decreased to levels 30-40% lower than controls at days 2 and 4 to rise again above control values on day 7 and 14. Atrial ir-ANP levels decreased at first both in the right and in the left atria with different latencies (1 and 2 days, respectively) and rose back towards control levels by day 4. Changes in the numerical density of specific granules followed a parallel temporal pattern. An increased in the individual volume of the granules followed was also observed. Investigation into the circulatory effects of T3 administration showed that the heart rate was increased by hour 12 after hormone administration (simultaneously with the early rise in plasma ir-ANP levels) and that blood pressure was increased by day 2.(ABSTRACT TRUNCATED AT 250 WORDS)


Nocturnal prolactin pulses in relation to luteinizing hormone and thyrotropin. The two hypothalamic releasing factors, luteinizing hormone releasing hormone (LHRH) and thyrotropin releasing hormone (TRH), have been shown to stimulate pituitary prolactin (PRL) release as well as their respective pituitary hormones, luteinizing hormone (LH) and thyrotropin (TSH). In this study the influence of LH and TSH regulatory mechanisms on nocturnal PRL secretion was investigated by evaluating whether the coincidence of PRL with LH and TSH pulses occurred more frequently than would be expected if the hormone generators were not coupled. Thirty night studies were conducted in twelve healthy male subjects. Six subjects underwent 3 studies and 6 subjects 2 studies. Blood was collected into aliquots at 10 min intervals throughout the night and plasma concentrations of PRL, TSH, and LH were determined. From the plasma profiles, hormone secretory rates were calculated using a method of deconvolution. Significant plasma and secretory hormone pulses were identified by a peak detection computer program. For statistical analysis the night studies of each subject were concatenated. Concomitance between the plasma pulses of both TSH and LH with PRL was insufficient to reject the null hypothesis of random coincidence. An increase in the number of subjects demonstrating significant coincidence between the hormone pulses was obtained when secretory pulses were analysed. Seven of the 12 and 10 of the 12 subjects showed significant concomitance between PRL and respectively TSH and LH. This proportion was sufficient to confirm copulsatility between PRL and LH. These results suggest that LH regulatory mechanisms are involved in the generation of the nocturnal pulsatile PRL profile, TRH may also play a role in the secretion of PRL at a central level, but was not reflected in the plasma or secretory profiles because of other overriding regulatory factors.


Visual evoked potentials in hypothyroid and hyperthyroid patients before and after achievement of euthyroidism. Thyroid hormone deficiency is frequently associated with central nervous system (CNS) disturbances such as mental retardation, convulsions, coma etc. Studies of quantitative changes in CNS in hypo- or hyperthyroidism are scarce. Evoked potentials is a good method of assessing the electrical response of the brain to different (visual, acoustic, somatosensory) stimuli and has been used extensively in the study of brain disturbances and to a lesser degree in metabolic diseases. We studied the visual evoked potentials (latency and amplitude) in 12 patients with hyperthyroidism and 15 patients with hypothyroidism, before treatment and after they became euthyroid. Four of the hyperthyroids (33%) had abnormally prolonged (> 104 msec) latencies before therapy. Two of them had clinical exophthalmos. No change was observed after euthyroidism was achieved. On the contrary 7 out of 15 (47%) hypothyroids had abnormally prolonged latencies which became normal in 4 when euthyroidism was achieved. Amplitude was lower than normal in 6 and became normal only in one of them after treatment. None of the hyperthyroid patients had amplitude changes. In conclusion, hypothyroid patients may have changes in the amplitude and/or the latency of visual evoked potentials which are reversible to a great extent with thyroxine. Evoked potentials is another method of studying in humans the metabolic effects of thyroxine deficiency in CNS.


Lack of any estrogenic effect of ipriflavone in postmenopausal women. Estrogen replacement therapy (ERT) has been demonstrated to prevent osteoporosis in postmenopausal women (PMW). However, several contraindications exist for ERT and many PMW cannot be treated. It has also been shown that too low doses of ERT are able to exert therapeutical effects on some climacteric symptoms but not on bone and compounds exerting synergic actions with ERT on bone without effects on other organs could be useful. The isoflavone derivative, ipriflavone, seems to have this effect but data are lacking on its endocrine effect in humans; thus, this study was undertaken to clarify in PMW whether ipriflavone exerts estrogenic activity. Evaluation of LH and FSH secretion during a 24-h period was performed in a group of 15 PMW after a single oral dose of 600 or 1,000 mg of ipriflavone or placebo, and after 7, 14 and 21 days of oral treatment with ipriflavone 600 mg and 1,000 mg/daily, administered in three divided doses. LH secretion was also evaluated during naloxone infusion before and after 21 days of ipriflavone, placebo or conjugated estrogen treatment (0.625 mg/day; CE). LH response to NAL treatment was absent during ipriflavone and placebo such as it was observed before treatments. By contrast, a significant increase of LH plasma levels was measured during naloxone infusion in CE-treated women. This result demonstrates that ipriflavone is unable to exert the same effects that estrogens do in PMW. In addition, no changes like in placebo group were seen on vaginal cytology in this group of subjects after 21 days, whereas a significant increase of superficial vaginal cells was observed after 21 days of CE treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Increased arterial and venous plasma noradrenaline levels in patients with primary hypothyroidism during hypothyroid as compared to euthyroid state. The use of venous plasma noradrenaline levels as a marker of general sympathetic tone has been questioned as changes in local sympathetic activity may influence the venous levels. To compare arterial and venous plasma noradrenaline levels in patients with primary hypothyroidism, arterial and venous blood were sampled during strictly standardized conditions during hypothyroid and euthyroid states. The patients were hospitalized for 5 days at a metabolic ward on a standardized sodium and potassium intake. On the fourth day catheters were positioned in the axillary artery and vein. Blood samples were drawn simultaneously for noradrenaline and adrenaline determinations during resting conditions. The arterial and venous plasma noradrenaline levels did not differ significantly, neither during hypothyroidism nor during euthyroidism. The arteriovenous difference in plasma adrenaline was similar during hypothyroidism compared to euthyroidism, indicating similar peripheral extraction rate of catecholamines during hypothyroidism as compared to euthyroidism. During hypothyroidism venous and arterial noradrenaline were significantly higher as compared to euthyroidism. In conclusion, there is no difference between arterial and venous noradrenaline levels either in the hypothyroid or the euthyroid state, and the peripheral extraction rate of plasma noradrenaline seems to be similar in hypothyroidism and euthyroidism. The local contribution of noradrenaline from the arm, reflecting local sympathetic nervous activity, is limited during resting conditions. In hypothyroid patients plasma noradrenaline levels are increased as compared to the euthyroid state, indicating increased general sympathetic activity in hypothyroidism.


Cervical pheochromocytoma: a rare localization and a difficult diagnosis. A 45-year-old hypertensive female with insulin-treated diabetes mellitus presented to our clinic with elevated urinary norepinephrine (NE) concentrations and a negative 131-metaiodobenzylguanidine (MIBG) scintigraphy, errouneously limited to the abdomen, for evaluation of a pheochromocytoma (Pheo). Despite antihypertensive medications blood pressure remained highly variable and frequently elevated. Further biochemical testing, including a glucagon provocation test and a clonidine-suppression test, revealed autonomous NE secretion. In order to avoid repeat MIBG-scintigraphy, other non invasive imaging techniques were performed, including real time sonography (7.5 MHz) of the neck which revealed a tumor. Fine needle aspiration of this tumor tissue demonstrated cells compatible with Pheo. Histology and immunohistochemistry of the excised tumor confirmed the diagnosis of Pheo. After surgical removal of the tumor, urinary and plasma NE levels normalized. Without any medication the blood pressure of the patient was now only slightly hypertensive. Only half of the daily insulin dose was needed to maintain the patient euglycemic.


Hypercalcemia associated with dysregulation of 1,25-dihydroxyvitamin D in arthritis. We describe an elderly man who presented with hypercalcemia associated with suppressed intact parathyroid hormone (PTH) levels. Despite renal insufficiency the circulating 1,25-dihydroxyvitamin D (1,25(OH)2D) was in the upper part of the normal range. Known causes of hypercalcemia were absent and mild hypercalcemia with suppression of intact PTH persisted until after bilateral hip replacement for severe arthritis (1 year after presentation). After hip replacement the ionized calcium normalized, intact PTH normalized, and 1,25(OH)2D decreased markedly. We believe the abnormalities in mineral homeostasis were related to production of 1,25(OH)2D by inflammatory mononuclear cells associated with arthritis.


Growth hormone deficiency throughout puberty. Growth during puberty does not appear to be the major determinate of final height in isolated GH deficient patients. Early diagnosis and commencement of therapy are probably the most important factors, as reflected by the correlation between final height and height at the onset of puberty. The cost effectiveness of increasing the dose of GH during puberty does not appear to represent any advantage from the data presently available. Indeed, such an approach may have a deleterious effect on final height by shortening the duration of pubertal maturation. Further prospective studies are required to demonstrate the effectiveness of manipulating the onset and duration of puberty using gonadotrophin releasing hormone analogues on final height in isolated GH deficiency.


[Fetal growth retardation: physiopathology. Review of the literature]. Fetal hypotrophy or intrauterine growth retardation is an important cause of fetal and neonatal mortality and a real cause of iatrogenic prematurity. When pregnancy is progressing normally there are histological changes in the spiral arteries with an equilibrium between the endothelial prostacyclin and platelet thromboxane on the one part and peroxides and Vitamin E on the other part. This allows uteroplacental circulation to flow rapidly and at a low pressure. Furthermore immunological tolerance of the mother's system in the relationship to her embryo plays a part in bringing about this special haemodynamic state. The physiopathology of retarded growth stemming from the blood vessels brings into play multiple mechanisms. The first cause is a defect in placentation which is shown up as insufficient trophoblastic invasion in the second stage. This brings about endothelial dysfunction with disappearance of the normal equilibrium of the prostaglandins and an alteration in the physiological equilibrium between the peroxides and a natural antagonist, which is vitamin E. Immunological disturbances have also been considered following studies on the immunohistochemistry and with the frequent association of obstetrical vascular complications together with phospholipid specific autoantibodies.


[The value of color Doppler ultrasound in the diagnosis of breast tumors]. A prospective study was performed in 102 patients, including 40 malignant tumors (13 subclinical), to evaluate colour Doppler in breast tumours. All patients had abnormalities that led to their selection for this study. 92 patients had a mammographic examination. 10% of malignant tumours were not detected by mammography. Mammographic abnormalities without any ultrasonographic confirmation were excluded from this series. All the lesions have been proven by cytology or histology. In the assessment of malignancy only one false negative and two false positive results were obtained. The sensitivity and specificity of colour Doppler are both of 97% with radiate vessels beneath the tumour and intratumoral vessels as malignant criteria. This non invasive method should be performed in lesions with tissue ultrasonographic appearances even before fine needle puncture. So it may be possible to select more surgical indications considering an ultrasonographic abnormality atypical, subclinic or post-therapeutic (radiotherapy or surgery).


[Immunologic aspects of endometriosis].

[Immunologic aspects of endometriosis]. Endometriosis is thought to result from implantation of endometrial tissue swept back into the pelvic cavity during menstruation, or from coelomic metaplasia of the peritoneum. Yet not all women with menstrual reflux develop endometriosis. The authors present and analyse the arguments in favour of immune system involvement in the physiopathology of this frequent disease. A knowledge of the antigens that have been recognized and of the operative mechanisms will probably make it possible to understand better the repercussions of endometriosis on fertility, to develop diagnostic methods less traumatic than those available at present and to modulate treatments and improve their effectiveness.


[Ovarian hyperstimulation syndrome during in vitro fertilization stimulations. Comparison between the experience of the Center for Medically Assisted Procreation of the Saint Antoine maternity department and a review of the literature]. The authors present their experience with the severe ovarian hyperstimulation syndrome when they were using stimulation for IVF and ET and they compare it with the literature. Over a period of 23 months. 154 stimulations were carried out using a long protocol associated with a CnRH agonist and hMG. 142 vaginal aspirations were carried out. The clinical pregnancy rate was 19% per aspiration and 21.9% per transfer. Of the 142 patients who were aspirated five had severe ovarian hyperstimulation with a favourable outcome. The authors suggest a physiopathological comparison for ovarian hyperstimulation and the management to be carried out when this complication occurs. They emphasise how potentially serious this complication is and that there has been no physiopathological explanation for its recrudescence since GnRH agonist have started to be used.


[Langerhans-cell histiocytosis of the vulva. General review and a case history]. Localisation of Langerhans' cell histiocytosis (HCL) is rare. In solitary form the prognosis for this pathology is good, if the tumour occurs in a woman of 36 years of age. (In this case the woman's age was 91) The appearances may be misleading clinically. The diagnosis is made by histological examination of P-S 100 positive cells and by finding Birbeck's granules on electron microscopy. Treatment may be surgical or as in our case medical.


[Hysterectomy and intraepithelial neoplasia of the lower female genital tract]. Delimiting the place of hysterectomy in cases of lower genital tract intraepithelial neoplasias in women. The laser and colposcopy centre (CCL) of a maternity unit in Lausanne in the Vaudois University Hospital Centre (CHUV). THE TYPE OF STUDY: Retrospective on 1,303 patients between 1986 and 1990. THE SUBJECTS AND TREATMENT: 853 cases of cervical intraepithelial neoplasia (CIN) and 79 cases of vaginal intraepithelial neoplasia (VAIN) were treated with CO2 laser. The situations in which hysterectomy could be considered in the course of treatment are discussed. They are: 1) Dysplasia persisting after treatment, 2) when pathological tissue is found on examining slides from conisation specimens, 3) micro-invasive carcinoma, 4) post-operative obstructive stenosis. The multiple location of dysplasia lesions of the lower genital tract was calculated for all the patients examined. It shows that hysterectomy itself will be insufficient to remove all dysplasias since frequently (9.2%) of lesions are found in the vagina in cases that have dysplasia of the cervix. Residual lesions after hysterectomy are shown up by VAIN which are responsible for the persistence of changes in the control smears (in which there were 9 cases of VAIN3 after hysterectomy in this series). Treatment therefore is hazardous and only poorly successful because the site of these lesions is often hidden in the scar through the top of the vagina. Furthermore their discovery in uncertain since there is a tendency at present to avoid out cytological screening of these patients once they have undergone hysterectomy. The high incidence of multifocal lesions and the possibility that is very real of residual dysplasia after hysterectomy has made the authors limit the place of hysterectomy in these cases, preferring to use conservative treatments and emphasizing the need to continue cytological controls for after treatment.


[Hematoma of the rectus abdominis muscle. Incident or accident of laparoscopy: A case report]. Laparoscopy can lead to complications in the abdominal wall, usually benign in outcome, including damage to an umbilical or epigastric vessel. A few simple rules can be drawn up to avoid (but not always) this type of accident. The treatment of this complication and the prognosis is discussed.


[Peritonitis due to a suppurating urachal cyst in a young woman. Review of the literature]. Peritonitis caused by a suppurating urachal cyst in a young woman. A review of the literature. Urachal cysts are not rare but occur much more often in children. In adults they may become infected and cause clinical symptoms including sub-umbilical mid-abdominal pain, fever, guarding and leukocytosis. Intraperitoneal rupture is rare (21 published cases) and only 7 cases have involved an adult female patient. The authors report the case of a 27 year old woman with peritonitis caused by rupture of a suppurating urachal cyst. 6 cm in diameter, for which the diagnosis had been established by pre-operative ultrasound. The surgical procedure carried out was total excision of the urachus, of the fascia between the umbilicus and the bladder, and partial excision of the dome of the bladder. Indeed total excision should have been carried out because of the risk of malignant degeneration of this mass to an adenocarcinoma.


[Cost effectiveness analysis of prenatal screening of trisomy 21 by maternal serum markers (hCG)]. Introduction of maternal serum markers for prenatal screening of Down's syndrome leads to a redefinition of the criteria used for identifying at risk women (i.e. in France, mainly based on maternal age of 38 and over). Effectiveness and costs of prenatal screening of Down's syndrome using such maternal serum markers will vary depending on the biological cut-off values defined, at each maternal age, to identify the population of pregnant women that will be sent to amniocentesis. On the basis of the first French prospective study of the use of human chorionic gonadotropin (hCG) measurement in maternal serum as a predictor of an increased risk of Down's syndrome, this paper shows that a screening policy combining maternal age with hCG measurement is more cost-effective than screening on the basis of maternal age alone. However, final decisions about hCG cut-off values should take into account the complex ethical dilemmas involved, especially the potential consequences of "false positives" and "false negatives", of this marker.


[Ectasia of the umbilical recess during the antenatal period. 3 case reports]. Having found three cases of ectasia of the umbilical recess in the antenatal period we decided to review the embryology and the ultrasound of the anatomy of the portal system. We consider that this is a normal anatomic variant of the portal system because the lesion is asymptomatic and regresses spontaneously.


[Ultrasonic diagnosis of a hereditary multiple malformation syndrome: Meckel-Gruber syndrome or Carpenter-Hunter syndrome]. Concerned by 2 cases of a recurring multiple fetal malformation syndrome in a consanguineous couple, the authors present the ultrasonic, clinical and paraclinical data that, when introduced into a computerised prenatal diagnostic programme, suggest a Meckel-Gruber or Carpenter-Hunter syndrome. The discovery of single or multiple fetal malformations requires not only complete echographic assessment, but also detailed post-abortum examination to allow optimal use of diagnostic aid programmes.


[Varicella in pregnancy after the 20th week of amenorrhea]. We report five cases of varicella pneumonia among ten otherwise healthy pregnant women who were admitted in our hospital between 1986 and 1991 with chickenpox. The precise frequency of this rare complication is not well known actually but analysis of the literature shows that the mortality rate is about 20%. Beside the problem of the fetal varicella syndrome, the other complication is the severe varicella of the neonate which can appear when varicella occurs in the mother within 5 days before, and 2 days after delivery. When primary varicella infection occurs during pregnancy clinical examination must be repeated for a week after occurring of the exanthema to find elements of severity significance. Acyclovir is the drug of choice (10 to 15 mg/kg every 8 hours) for 7 days when pneumonia is present. Varicella-zoster immunoglobulin is useful for prophylaxis and for neonates with high risk of severe varicella.


[Prematurity due to medical decisions. Analysis of a retrospective study of 18 maternity hospitals in the periphery of Paris]. We have analysed induced prematurity in a retrospective study of 18 maternity hospitals located in the area of Paris: the Ile-de-France. The rate of induced prematurity was 1.4%, whereas the total rate of prematurity in our study was of 4.6% for a total number of 16,337 deliveries. The great majority of those medically induced prematurity was decided at and after 33 weeks, and the main cause was intrauterine growth retardation. Caesarean section was performed in 60% of the cases.


[Cerebral aneurysms and pregnancy: 4 cases]. Haemorrhagic cerebral accidents are the commonest neurosurgical diagnoses made in pregnancy. The state of pregnancy makes it more likely that an arterial or an arteriovenous aneurysm will rupture and this is the principal cause of most haemorrhages. They occur more often in primiparae in the third trimester of pregnancy. The clinical picture is classical. The conformation of the diagnosis is made by scanning and angiography. The main differential diagnosis is eclampsia. Neurosurgical treatment should be carried out immediately whenever possible in order to avoid the two great risks that follow, namely recurrence of haemorrhage and secondly ischaemia. As far as the obstetric side is concerned, Caesarean section would only be indicated if: the clinical state of the mother is severe with coma and brain stem damage when the child is viable, if there is symptomatic vascular malformation diagnosed at term, if there is haemorrhagic arteriovenous malformation which is highly liable to occur and cannot be operated on without risks for the child if viable, if, finally, the interval between the surgical treatment of the condition and labour is less than 8 days. In all other cases a vaginal delivery is preferable under epidural anaesthetic which should be given if medical induction is carried out, and where instrumental delivery is being carried out systematically, unless radical treatment is being performed. The prognosis which is, in spite of all steps that may be taken, poor, depends on the initial neurosurgical stage and the nature of the causes of lesion and the possibilities of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


[Hydrocephalus with cerebrospinal fluid shunts and pregnancy: 2 cases]. During pregnancy 50% of all cases with a ventriculo-peritoneal shunt malfunction. This is because of anatomo-physiological changes associated with the pregnant state, and shows itself as a rise in intracranial tension. There were no acute neurological complications at term; with the malfunction of the shunt distally, vaginal delivery is preferable. It is not necessary routinely to carry out instrumental delivery.


[Sub-capsular hematoma of the liver during pregnancy: a case report]. A sub-capsular haematoma of the liver is a rare but very serious complication of pregnancy. We report a case of a primipara whose pregnancy was first complicated by hypertension, then by a haematoma of the liver at the 28th week of amenorrhoea. It was diagnosed because of the clinical symptoms associated with the hypertension and a pain in the right hypochondrium and nausea. The diagnosis of haematoma of the liver was made before rupture and confirmed by ultrasound of the liver and by tomodensitometry made it possible to avoid operating on the liver during the laparotomy that was carried out to terminate the pregnancy. Regular X-ray follow-ups showed that the lesion had regressed in six months. MRI investigation of the parenchyma of the liver showed no cause of the lesions. If there is even the slightest suspicion of a haematoma of the liver in pregnancy, ultrasound nowadays should be carried out because it is indispensable to make the diagnosis.