[How I treat a neonatal Herpes simplex virus infection]. / Comment je traite l'infection néonatale à Herpès simplex.

Neonatal herpes simplex virus infection is rare but important to recognize because of the major risk of sequelae or death. The diagnosis is mainly based on specific clinical and biological analyses. Aciclovir is the treatment of choice, duration of administration depending on the severity of the disease. A six-month treatment with suppressive-dose oral aciclovir is recommended to improve the child's prognosis. From a clinical case, we reviewed the literature to improve the management.

L'infection néonatale à Herpès est peu fréquente mais importante à reconnaître vu le risque important de séquelles ou de mortalité. Le diagnostic repose principalement sur des analyses cliniques et biologiques spécifiques. L'aciclovir est le traitement de choix, la durée d'administration varie en fonction de l'importance de l'atteinte. Un traitement de 6 mois par voie orale est conseillé pour améliorer le pronostic de l'enfant. A partir d'un cas clinique, nous avons revu la littérature pour connaître les dernières recommandations afin d'améliorer la prise en charge.

Loss of the common "A" determinant of hepatitis B surface antigen by a vaccine-induced escape mutant.

A previous study (Carman, W. F., A. R. Zanetti, P. Karayiannis, J. A. Waters, G. Manzillo, E. Tanzi, A. J. Zuckerman, and H. C. Thomas. 1990. Lancet. 336:325-329) demonstrated a variant hepatitis B surface antigen (HBsAg) in a vaccinated child born to a hepatitis B virus-infected mother. A substitution of arginine for glycine at amino acid 145 in HBsAg was observed. In this study the effect of this substitution on the common "a" determinant of this protein, against which protective immunity is directed, is investigated. Using recombinant HBsAg with and without the amino acid substitution, the binding of monoclonal antibodies that recognize different epitopes of the "a" determinant, was shown to be destroyed by the presence of arginine at amino acid 145. In convalescent and vaccinee sera, antibody binding to HBsAg was not inhibited by the variant HBsAg. Immunization with the variant HBsAg, although eliciting a high titer antibody that recognized the variant, produced a low titer of antibody recognizing the native protein. Studies in mice demonstrate that the immunogenicity of the variant protein is also substantially altered. The data presented here demonstrate that this variant evades the known protective anti-HBs response and lends support to the suggestion that this mutation arose as the result of immune pressure.

Protein-chemical analysis of pertussis toxin reveals homology between the subunits S2 and S3, between S1 and the A chains of enterotoxins of Vibrio cholerae and Escherichia coli and identifies S2 as the haptoglobin-binding subunit.

The purified toxin of Bordetella pertussis was dissociated in 5 M urea in the presence of immobilized haptoglobin. The toxin was dissociated in free S1, free S5 and the free complexes S2-S4 and S3-S4, with S2-S4 as the only haptoglobin-binding moiety, identifying S2 as the haptoglobin-binding protein. Partial NH2-terminal amino acid sequences were obtained from the dissimilar toxin subunits, after separation by SDS-polyacrylamide gel electrophoresis followed by electroblotting onto polybrene-coated glass-fiber sheets. The sequences reveal extensive homology of the N-terminal portions of the constitutive subunits S2 and S3 and between S1 and the enterotoxin A chains of Vibrio cholerae and Escherichia coli.

Purification and properties of an endo-beta-1,4-glucanase from Clostridium thermocellum.

The extracellular cellulolytic enzymes of the thermophilic anaerobe Clostridium thermocellum occur as a protein complex or aggregate which, until now, has not been resolved into individual enzyme components. By using QAE-Sephadex A50 chromatography in the presence of 6 M urea, it was possible to split the complex into distinct protein fractions. One of these fractions contained an endo-beta-1,4-glucanase which was isolated at a high degree of purity and was identified by its ability to hydrolyze trinitrophenylated carboxymethylcellulose. The enzyme is of monomeric nature, with a molecular weight of 56,000. It has an isoelectric pH of 6.2 and an optimum pH of 6.0. It hydrolyzed carboxymethylcellulose and, at a slower rate, cellulose powder. The major end products of cellulose degradation are glucose, cellobiose and cellotriose; cellotetrose is formed as an intermediate product. No specific small molecular weight activator or inhibitor was found except cellobiose and, to a lesser extent, glucose, which at high concentrations partially inhibit the activity of the enzyme. The temperature dependence of the enzyme is related to the thermophilic character of the producing microorganism.

Immunization of mice by recombinant OspA preparations and protection against Borrelia burgdorferi infection induced by Ixodes ricinus tick bites.

The wide distribution of Borrelia burgdorferi, the spirochete causing Lyme borreliosis, represents a human health hazard in many areas of the world. Vaccination has been proposed as an effective prevention strategy. Vaccination experiments were conducted with preparations of recombinant outer surface protein A (OspA) derived from Borrelia burgdorferi strain ZS7. Mice received three doses (1 microgram each) of the antigens adsorbed to aluminum hydroxide. A strong immune response to the vaccine antigen was observed. Mice were challenged after immunization, using Ixodes ricinus nymphal ticks infected with Borrelia burgdorferi strain ZS7. Infection was investigated by ear biopsy culture, xenodiagnosis with uninfected larvae and serological response to Borrelia burgdorferi antigens. All unimmunized control animals were found to be infected, while all immunized animals were found to be protected against infection by Borrelia burgdorferi. In addition, most adult ticks derived from nymphs that fed on immunized mice were found to be free of spirochetes.

Properties of a recombinant yeast-derived hepatitis B surface antigen containing S, preS2 and preS1 antigenic domains.

Yeast cells have been engineered to express mixed HBsAg particles containing the S and a modified large (L*) protein. Their construction resulted in reduced protease sensitivity, reduced glycosylation and complete inactivation of the polymerized human albumin binding site. The particles exposed the S, preS1 and preS2 antigenic determinants and induced an immune response against the three domains. Highly purified preparations have been obtained and are presently being tested in human volunteers.

Automated control of postoperative hypertension: a prospective, randomized multicenter study.

Hypertension after a cardiac operation is a frequent phenomenon. Complications resulting from this include bleeding, disruption of vascular suture lines, subendocardial ischemia, and possible cerebrovascular accidents. Treatment with sodium nitroprusside has become accepted practice to prevent these complications. To improve control of arterial blood pressure, a closed-loop system for sodium nitroprusside administration was developed. A prospective, randomized multicenter study was carried out postoperatively in 180 cardiac surgical patients to evaluate the performance of this system compared with manual control of infusion. Adherence of mean arterial blood pressure to +/- 10% of the target blood pressure occurred 85% of the time with the automatic system and 61% of the time with manual regulation (p less than 0.0001). With the automatic system, there was less hypertension (9% versus 22%; p less than 0.0001) and hypotension (6% versus 22%; p less than 0.0001). The superior control of hypertension was achieved more rapidly with less requirement for nurse regulation of infusion rate. The superior control of blood pressure resulted in less chest tube drainage in the automatic mode (720 mL versus 840 mL; p less than 0.05).

On-line biomass monitoring by capacitance measurement.

An in-situ, steam-sterilizable capacitance probe was used to follow the biomass concentration on-line, in bioreactors from 20 to 2000 l total volume. Microbial cultures of Saccharomyces cerevisiae, Pichia pastoris and Streptomyces virginiae were grown in batch and fed-batch culture in both defined and complex media in order to demonstrate the wide dynamic operating range of the instrument. A linear correlation was found between the on-line capacitance measurement and the off-line measurements (optical density, OD620; packed mycelial volume, PMV; biomass concentration X, and colony forming units, CFU ml-1) for biomass concentrations (dry cell weight) up to 30 g l-1 (St. virginiae), 106 g l-1 (S. cerevisiae) and 89 g l-1 (P. pastoris). The on-line capacitance measurement was slightly influenced by variations in agitation speed and strong extraneous radio frequencies. A specific capacitance constant (Cs) was defined for all microbial cells which was dependent on cell viability and cell size. The Cs was easy to calculate using the on-line capacitance measurement and an off-line estimation of biomass concentration. The Biomass Monitor proved suitable for precise on-line monitoring of both homogeneous (uni-cellular) and heterogeneous (mycelial) cultures in bioreactors.

Estimation of current leakage in left and right ventricular conductance volumetry using a dynamic finite element model.

Leakage of electric current through cardiac structures surrounding the ventricle is a primary source of error during ventricular volume measurements using a conductance catheter. This error can be represented as a leakage volume, VL. VL is generally estimated by a saline-bolus method, and is assumed constant throughout the cardiac cycle. However, dynamic changes in ventricular volume and cardiac wall thickness could change VL. To estimate VL, a dynamic finite element model of the heart was developed based on MR images. Conductance measurements were simulated using a modeled conductance catheter, and true VL was calculated. VL varied from 22.7 ml (end-systole) to 26.4 ml (end-diastole) in the left ventricle and from 19.9 ml (end-systole) to 26.9 ml (end-diastole) in the right ventricle. The saline-bolus method underestimated VL in both the left (VL = 19.4 ml) and the right (VL = 4.1 ml) ventricular volume measurements. VL increased linearly with the ratio of blood to tissue resistivity, and changed minimally with catheter position. These results indicate that VL has to be estimated dynamically throughout the cardiac cycle to obtain accurate cardiac volume measurements. The results also show that the saline bolus method does not estimate current leakage accurately, especially in the right ventricular volume measurement.

Real-time continuous measurement of right ventricular volume using a conductance catheter.

The authors propose using a multi-electrode conductance catheter to measure continuous right ventricular volume. True ventricular volume measurements are affected by four main sources of error. 1) field non-uniformity, 2) catheter curvature, 3) blood conductivity changes, and 4) leakage of current through surrounding tissues. Three-dimensional finite-element models were developed to investigate the effects of these sources of error and to devise schemes for correcting them. The models include an axisymmetric cylindrical model, a rectangular block model, and a heart model with left and right ventricular chambers. The heart model is built from conical primitives, with major dimensions derived from the literature. Finite-element simulations showed that volume measurements were underestimated due to field nonuniformity to as much as 1/25th actual volume in segments near the exciting electrodes. The extent of underestimation in a segment decreased with increasing distance of the segment from the exciting electrodes and increased for larger segmental volumes. Catheter curvature overestimated measured volume by as much as 4.5 times when the curvature was increased from 0.0 to 1.25 (from a straight catheter to a very curved one). The leakage of current through surrounding tissues overestimated volume by nearly 30%. The sensitivity of volume measurement to blood resistivity changes was found to be very high, at 70%. Correction factors established with the computer models compensate for field nonuniformity. Mathematical mapping of the curved catheter onto a fictitious straight catheter corrects for the catheter curvature error. Correction for both nonuniform field and catheter curvature allowed measurement of total ventricular volume with an error of 7%. Leakage current is determined by using different frequencies to build the catheter electric field and to separate tissue and blood resistance paths. Using this scheme, the percentage overestimation in volume measurement due to leakage could be determined with an accuracy of 85%. The proposed correction scheme for blood conductivity changes involves the in-vivo measurement of blood conductivity with the catheter itself. It was found that blood conductivity could be determined with insignificant error (< 0.5%) so long as the blood volume around the exciting electrodes had a radius of more than the electrode spacing.

Cloning of the bluetongue virus L3 gene.

The genes of the bluetongue virus (BTV) serotype 17 have been cloned into pBR322 by tailing both strands of the double-stranded RNA with polyadenylic acid, transcribing them with reverse transcriptase with an oligodeoxythymidylic acid primer, hybridizing the cDNA products, and completing them into duplex structures with the Klenow fragment of Escherichia coli DNA polymerase. After cloning the double-stranded cDNA molecules into pBR322, the complete sequence of the cloned L3 gene was determined. The clone is 2,772 nucleotides long (1.78 X 10(6) daltons), excluding the 3' polyadenylic acid sequence, and has an open reading frame which codes for a protein of some 901 amino acids (103,412 daltons). This clone can hybridize L3 RNA segments of three other U.S. BTV serotypes, BTV-10, -11, and -13 in addition to -17 but not the equivalent RNA segment of epizootic hemorrhagic disease virus of deer, an orbivirus related to BTV.

Cloning of DNA sequences encoding foreign peptides and their expression in the K88 pili.

A genetic system that allows the cloning of a peptide-coding sequence in the Escherichia coli K88ac and K88ad pilin genes and their expression as recombinant pili has been constructed. Two insertion vectors were created by subcloning the pilin genes in a pBR322 plasmid and replacing the coding sequence of two nonconserved clusters by a linker. The K88ac helper genes were subcloned in the compatible pACYC184 plasmid, and expression of pili by bacteria carrying both plasmids occurred by complementation. Two peptide-coding sequences of the influenza hemagglutinin were cloned in both insertion vectors, and recombinant pilins were shown to be assembled in pili. One recombinant pilus was shown to elicit antibodies against the synthetic peptide in immunized rats. The somatostatin-coding sequence was cloned in both vectors and led in one case to detectable pilus production. The fused somatostatin was shown to be recognized by specific monoclonal and polyclonal antibodies.