Gene transfer of anti-gp41 antibody and CD4 immunoadhesin strongly reduces the HIV-1 load in humanized severe combined immunodeficient mice.

OBJECTIVE: To study the anti-HIV-1 effects of the delivery of anti-gp41 monoclonal antibody (mAb) and soluble CD4 (sCD4) immunoadhesin by genetically modified cells in HIV-1-infected, humanized severe combined immunodeficient (SCID) mice. DESIGN: The complementary DNA of mAb 2F5, an anti-HIV-1 gp41 antibody, and of sCD4-IgG chimeric immunoadhesin were transferred into 3T3 cells using Moloney murine leukaemia virus vectors. The cells were then incorporated into a collagen structure called the neo-organ, which allowed the continuous production of the therapeutic molecules. METHODS: The antiviral effects in vivo of 2F5 or sCD4-IgG or both compounds were evaluated in neo-organ-implanted SCID mice that were grafted with human CD4 CEM T cells and challenged with HIV-1 Lai or MN. RESULTS: In SCID mice implanted with 2F5 neo-organs, antibody plasma levels reached 500-2000 ng/ml. Viral loads after HIV-1 challenge were significantly reduced in neo-organ-implanted HIV-infected mice. Although 29 x 10(7) and 13 x 10(8) HIV-1-RNA copies/ml were detected at 12 days in the controls (mice injected with Lai and MN, respectively) less than 16.5 x 10(3) HIV-1-RNA copies/ml were observed in all implanted mice injected with either Lai or MN. The intracellular viral load was also reduced in CD4 cells recovered from the implanted mice. Comparable antiviral effects were obtained with CD4-IgG neo-organs. CONCLUSION: Our results confirm the anti-HIV properties of 2F5 and sCD4-IgG continuously produced in vivo after ex-vivo gene therapy in SCID mice.

Experimental gene therapy: the transfer of Tat-inducible interferon genes protects human cells against HIV-1 challenge in vitro and in vivo in severe combined immunodeficient mice.

OBJECTIVES: To evaluate in vitro and in vivo a strategy for gene therapy for AIDS based on the transfer on interferon (IFN)-alpha, -beta and -gamma genes to human cells. DESIGN: Human U937 promonocytic cells were stably transfected with Tat-inducible IFN expression vectors conferring an antiviral state against infection with HIV. METHODS: Transfected cells were either infected by HIV-1 in vitro or transplanted into severe combined immunodeficient (SCID) mice for an HIV challenge in vivo. RESULTS: U937 cell lines stably carrying IFN transgenes under the positive control of the HIV-1 Tat protein were highly resistant to HIV-1 replication in vitro. This antiviral resistance was associated with a strong induction of IFN synthesis immediately following the viral infection. HIV-1 proteins were found to be specifically trapped within the genetically modified cells. In contrast, all IFN-U937 cells permitted full HIV-2 replication. Transfected cells injected into SCID mice and challenged against HIV-1 were strongly resistant to infection when cells were transduced with IFN-alpha of IFN-beta genes. However, IFN-gamma-transfected cells permitted HIV-1 infection in vivo despite the induction of a high level of IFN-gamma secretion. The quantity of proviral DNA was 10(5)-fold lower in IFN-alpha- or IFN-beta-transfected U937 cells collected from these SCID mice than that in non-transfected cells. CONCLUSIONS: Our results substantiated the validity of a strategy, bases on the transfer of HIV-1-inducible IFN-alpha or IFN-beta genes, to confer antiviral resistance to human cells.

Prenatal diagnosis of Walker-Warburg syndrome in three sibs.

Walker-Warburg syndrome (WWS) is an autosomal recessive condition characterized by diffuse neurodysplasia, resulting in brain and eye abnormalities. We report on 3 prenatally diagnosed cases of this syndrome born to a consanguineous couple. An ultrasonographic examination showed hydrocephalus at the 27th week of the first pregnancy. Amniocentesis documented a normal male karyotype. The couple opted for termination of the pregnancy but declined an autopsy. Seven months later, hydrocephalus was observed at 20 weeks of the second pregnancy. Termination of pregnancy was performed at the 22nd week. Autopsy of this male fetus showed dilated ventricles, thin cortex, and type II lissencephaly with microscopic evidence of chaotic architecture. Eye examination showed retinal dysplasia. Notwithstanding the lack of demonstrable muscle change, the diagnosis of Walker-Warburg syndrome was made. Ten months later, hydrocephalus was discovered in the third fetus, a female, at 13 weeks of gestation. Termination of pregnancy was performed at 20 weeks. At autopsy, brain, eye, and muscular findings were similar to those of the previous case. In addition, cystic changes and a stenosis of the pyelo-ureteral junction were found in the right kidney. Type II lissencephaly and retinal dysplasia are characteristic of WWS. Muscular dystrophy has been pointed out as an additional abnormality in postnatal cases. By contrast, the lack of demonstrable muscle changes in the fetal period must be emphasized. Those cases illustrate practical problems in the ultrasound and pathologic diagnosis of WWS in the fetal period.

High-risk genotype for type 1 diabetes: a new simple microtiter plate-based ELOSA assay.

The main contribution to genetic susceptibility for type 1 diabetes (T1D) is conferred by the HLA class II genes, with a major involvement of the DQB1*02 and 0302 alleles. The aim of our study was to develop a simple and rapid method suitable for identifying individuals with an HLA-associated T1D risk using whole blood as a source of DNA and reverse hybridization on microtiter plates (ELOSA). DNA was extracted from whole blood using various extraction methods. The PCR-amplified second exon of the DQB1 gene was hybridized at 37 degrees C for 1 hr to a set of 11 capture probes immobilized on a microtiter plate (eight-well strip per test) and corresponding to T1D susceptibility (S), protection (P), or neutral (N) alleles. Colorimetric analysis was then performed using specific oligonucleotides coupled to horseradish peroxidase and OrthoPhenyl Peroxidase (OPD) substrate. DNA samples corresponding to French (Rhône-Alpes area) T1D patients (n = 128) have been genotyped with the HLA-T1D prototype. A strong correlation is observed between susceptible genotypes and the disease, because 92.2% of the T1D individuals screened have at least one susceptible allele (DQB1*02 or *0302), thereby strengthening interest in analyzing DQB1 alleles as HLA-linked T1D markers in our Rhône-Alpes area population. Interestingly, clear T1D-associated genotyping results have been observed when using DNA samples extracted from dried blood spots, making it possible to envisage such genotyping in geographically dispersed affected families, for large-scale newborn screening, and for the inclusion of high-risk patients in clinical trials aimed at preventing the disease.

The diagnosis of carpal tunnel syndrome. Sensitivity and specificity of some clinical and electrophysiological tests.

The study group consisted of 100 persons referred with suspected carpal tunnel syndrome. Clinical and neurophysiological examinations were performed blinded from each other. The gold standard for the carpal tunnel syndrome (CTS) diagnosis was based on the results of these examinations but relief of CTS symptoms after surgery was also required. The sensitivity and specificity for the combined results of the clinical examinations were 94% and 80% respectively, and for the neurophysiological examinations, 85% and 87%. Of the neurophysiological methods used, the quotient of sensory nerve conduction velocity between palm to wrist and wrist to elbow was best and the cut-off for this test was studied by means of an ROC-curve. According to our results clinical examination by an experienced doctor seems to be sufficient if there are typical symptoms of carpal tunnel syndrome, but if there is a history of pain, atypical symptoms or earlier fractures in the arm, wrist or hand, it is important to add a neurophysiological examination.

[In vitro and in vivo inhibition of HIV1 replication by retroviral transfer of interferon alpha, beta, or gamma genes: application to gene therapy of AIDS]. / Inhibition in vitro et in vivo de la réplication du VIH1 par transfert rétroviral des gènes d'interféron alpha, beta ou gamma: application à la thérapie génique du sida.

Somatic gene therapy is defined as the transfer of a heterologous gene into an organism for the purpose of correcting a genetic defect or providing a new therapeutic function to the target cell and thus inducing a cure or improving associated symptoms. While encouraging results have been generated by recent clinical evaluation of combination of anti-viral drugs, Aids still constitute an obvious candidate among the infectious diseases which might be treated by gene therapy. We have therefore chosen to develop and evaluate a gene therapy strategy based on the transfer into human target cells of HIV1-inducible interferon (IFN) alpha, beta or gamma genes. In a preliminary study, myeloïd U937 cell lines transfected with expression vectors containing the IFN alpha, beta or gamma genes under the control of the long terminal repeat (LTR) sequences of HIV1 were shown to be strongly resistant against an in vitro and in vivo (in HIV1 challenged SCID mice model) HIV1 infection. This cellular resistance was correlated with a strong induction of transgenic IFN synthesis and for IFN gamma, with a defect of HIV particles maturation. Secondly, construction and production of high titer retroviral vectors containing Tat-inducible IFN genes allowed efficient transduction of lymphoïd cell lines and human primary lymphocytes. These transduced cells were shown to be highly resistant against laboratory and primary HIV isolates. Taken together, our in vitro and in vivo results suggest that HIV1 inducible IFN gene therapy can be beneficial to HIV-infected individuals provided the fact that methods are developed that allow the efficient transduction of human hematopoïetic stem cells.

The serum reserve albumin concentration for monoacetyldiaminodiphenyl sulphone and auditory evoked responses during neonatal hyperbilirubinaemia.

Auditory brainstem evoked responses (ABR) were recorded in 9 neonates with hyperbilirubinaemia. Pathological recordings were found in two children showing absence of waves and prolonged latencies. There was no correlation between latencies to waves and the total serum bilirubin concentration. The serum reserve albumin concentration for monoacetyldiaminodiphenyl sulphone (MADDS) was, however, inversely related to the latencies in the ABR recordings. Our findings suggest that the binding properties of serum albumin contribute to the risk of bilirubin toxicity and that, in this study, the reserve albumin concentration for MADDS seemed to be of greater significance than the total bilirubin concentration.

Effects of a behavioral intervention on epileptic seizure behavior and paroxysmal activity: a systematic replication of three cases of children with intractable epilepsy.

Three children with very frequent refractory epileptic seizures underwent a behavioral intervention consisting of symptom discrimination, countermeasures, contingent relaxation, and positive reinforcement for correct responses in a systematic replication series. The studies involved a 6-h nonintervention base rate, a 6-h treatment phase, and a 6-h nonintervention follow-up under laboratory conditions for each child. Neurophysiologic and behavioral measures of the effects of treatment were made using electroencephalogram (EEG)-video equipment. Effects of treatment were assessed by using a random sample of EEG-video sequences in base rate and follow-up. Results showed that no significant reduction of either seizure behavior or paroxysmal EEG activity was found subsequent to training in discrimination of early paroxysmal activity and/or sensations preceding seizures. Both seizure behavior and paroxysmal activity were significantly reduced in all three cases following intervention with an adapted countermeasure technique. No additional effects could be noted subsequent to the application of either contingent relaxation or positive reinforcement for correct responses. Paroxysmal EEG changes and seizure behavior were highly correlated. Reduction of the clinical manifestation or seizure response by behavioral manipulation was accompanied by a reduction of the total amount of paroxysmal activity as measured by the EEG.

Effect of dorsal column stimulation on pain-induced intracerebral impulse patterns.

Evoked electrical potentials were recorded via intracerebral electrodes in a patient with stump and phantom limb pain who had a previously implanted dorsal column stimulator. When pain was elicited by peripheral stimulation it was found that positive deflections appeared in the ventrolateral nucleus of the thalamus at time latencies corresponding to the propagation velocities of A delta- and C-fibres. Dorsal column stimulation completely eliminated the C-fibres. Dorsal column stimulation completely eliminated the C-fibre deflection and partially eliminated the A delta-fibre deflection.

Epidemiology of neuromuscular diseases, including the postpolio sequelae, in a Swedish county.

The epidemiology of neuromuscular diseases was studied in the county of Orebro, Sweden (study population 270,000). Several different sources of data were utilized, compared and validated. On the prevalence of day (January 1, 1988) 474 patients were identified. The rate per 100,000 population was 92 for the postpolio sequelae (PPS) and 84 for the other neuromuscular diseases (motor neuron disease 9, hereditary neuropathies 9, myoneural disorders 16, myotonic disorders 19, muscular dystrophies 20 and myositis 11). Of the patients with the PPS, 80% reported late-onset symptoms. On the basis of an expanded survey including all medical records in one health care district, the prevalence of the PPS was estimated to be 186/100,000 population.

Motor blockade and EMG recordings in epidural anaesthesia. A comparison between mepivacaine 2%, bupivacaine 0.5% and etidocaine 1.5%.

In a double-blind study young volunteers randomly received 20 ml of mepivacaine 2%, bupivacaine 0.5% or etidocaine 1.5% epidurally, all solutions with adrenaline. The mean cephalad spread of pin-prick analgesia was equal (T10) in the groups, but the duration was longest for bupivacaine and etidocaine. The motor blockade of the rectus abdominis muscles was assessed quantitatively by rectified integrated electromyographic recordings (RIEMG) and as number of turns in EMG recordings [changes in the direction (rise/fall) of the EMG; TURNS] from three different segmental levels, T7, T9 and T11. The motor blockade of the quadriceps muscles was estimated by EMG recordings simultaneously with muscle force measurements at maximal isometric knee extension. Motor blockade was also evaluated by the Bromage scale. There was good correlation (correlation coefficient 0.91) between RIEMG values and muscle force in knee extension during epidural anaesthesia. TURNS showed a non-linear relationship to isometric force during epidural anaesthesia and added no further information. At the lower parts of the abdomen (T11), etidocaine gave more profound and longer motor blockade than mepivacaine. For quadriceps muscle function, motor blockade was almost complete with all three local anaesthetics; the duration of maximum motor blockade was short (45-60 min) for mepivacaine, but about 5 h with etidocaine. At the time when the Bromage scale indicated complete regression of motor blockade, the muscle force of knee extension was only 30% and the quadriceps RIEMG 35% of control values and 1-3 h remained until the time of mobilization.(ABSTRACT TRUNCATED AT 250 WORDS)

F response and H reflex for monitoring nerve block during epidural analgesia with ropivacaine.

The effect of a single epidural injection of ropivacaine on the motor and sensory function controlled from the L5/S1 level was investigated in 28 male volunteers. Concentrations of 1%, 0.75%, or 0.5% ropivacaine, 20 mL, administered at the L2/3 level were studied. Motor function was assessed quantitatively (measurement of muscle force by mechano-transducers), and sensory function by the pinprick method. In addition, F response and H reflex, tests which measure the conduction velocity in the central parts of peripheral nerves, were used. Epidural ropivacaine caused dose-dependent prolongation of the latencies of both these variables. F response latency recovered significantly later than motor function measured by mechano-transducers in the two lower concentration groups. H reflex latency recovered significantly later than sensory function assessed by the pinprick method in all three concentration groups. The time needed for recovery of F and H latencies was not significantly longer than the time from epidural injection to mobilization. At the time when the subjects could go through the mobilization procedure, 12 of 28 subjects were not completely recovered. In 5 of these 12 subjects, the H reflex latency was persistently prolonged at the end of the investigation, long after the subjects felt "normal" again. On follow-up recordings 5 mo later, the baseline latency had been regained in all five subjects. We conclude that F response and H reflex latencies are good indicators of the inhibition of nerve impulse conduction induced by epidural analgesia.

[An epidemiological investigation of pregnancy toxemia]. / Etude épidémiologique de la gestose ou dysgravidie tardive.

Importance of various factors: biometric, social, ethnical and medical, in the development of late gestosis has been assessed by the authors. Their bearing on the course of gestosis and the child's condition at birth has also been evaluated. The study has been conducted in 500 cases of dysgravidity. Some conclusions could be drawn: The high incidence of the disease in adolescent girls, primiparae and also women over 35 years of age who had multiple pregnancies. The impact of social factors. The unfavourable effect of obesity and even more so of exagerate weight increase during pregnancy. This latter fact is in contrast with the severity of lean forms developing in women with frail built. The need to increase the number of antenatal consultations if true prophylaxis is to be secured. In any case here is a direct relationship between the impact of the disease on the fetus and its severity.

[The interest of the plasma estrone + estradiol/estriol relationship in the surveillance of threatened premature birth, late pre-eclampsia and prolonged pregnancy]. / Intérêt du rapport oestrone + oestradiol/oestriol plasmatique dans la surveillance des menaces d'accouchement prématuré des gestoses tardives et des grossesses prolongées.

Although fully aware of the limits of their method, the authors consider the study of estrone + estradiol/estriol relationship, all these hormones being assayed in the mother's plasma, as a valuable aid in the difficult surveillance of threatened premature birth, late gestosis and prolonged pregnancy. Indeed, when the value of the relationship shows to be below I, the threatened premature delivery has nearly always a favourable course toward its resolution. In late gestosis, the rate of the relationship satisfactorily correlates with the gestosis index of Rippmann and Goecke, thus permitting to establish true prognosis. Finally, in prolonged pregnancy a relationship below I nearly always indicates good fetal condition.

[Plasma estriol in the surveillance of the second half of pregnancy. Report of 227 cases]. / L'oestriol plasmatique. Son dosage par une méthode chimique, son intérêt dans la surveillance de la seconde moitié de la grossesse. A propos de 227 dosages.

The authors have attempted to devise a simple technique based in exclusively chemical methods to assay plasma estriol. The results obtained have shown the lack of fiability of such a method, in spite of fairly good correlation with urine estriol levels. Only methods utilizing labelled estrogen or radiomyological techniques can yield fiable results in the assay of plasma estriol.

[Major lesions of the testicles observed in infants born by breech presentation]. / Lésions majeures des testicules observées chez des enfants nés par le siège.

Surgical investigations of the scrotom were carried out on six infants born by breech presentation. Three of them exhibited clinically significant oedema of the scrotum with haematic infiltration. In two others a bilateral haematoma of the scrotum and of the distal extremity of the penis was noted. In the remaining infant the scrotum was normal at birth but a few hours later an increase in volume and an induration were noted on one side. During surgery the authors noted an intravaginal torsion of the testicle with necrosis confirmed histologically. In four cases there was infiltration of blood into the testicle and the spermatic cord histological examination showing separation of the seminiferous tubules by an effusion of blood, the vegetative cells and the gonocytes being intact. In the last case the testicles and the spermatic cords were normal. The indications for operation on the scrotum were based on opacity to transillumination. The prognosis varies in relation to the lesions discovered. Atrophy of the testicles follows after six hours in cases of torsion while the evolution appears to be better in cases of haematic infiltrations. Breech presentation leads, in a certain number of cases, to haematic infiltration of the testicles and spermatic cords and seems to be a factor leading to torsion of the testicles. Detailed and repeated examinations of the scrotum are thus indicated after births of this type.

Evaluation of motor blockade by isometric force measurement and electromyographic recording during epidural anaesthesia--a methodological study.

Methods for assessing motor blockade by means of isometric force measurements and surface electromyographic (EMG) recordings in the lower extremities and abdominal wall were evaluated in 30 volunteers. The coefficients of variation were 10% for force measurements and 14% and 20% for average rectified EMG (RIEMG) recordings over the quadriceps muscle and abdominal muscles, respectively, and 8% overall for TURNS (the number of changes in the sign of the direction of the EMG signal). Seven of the 30 volunteers received epidural anaesthesia with 20 ml of mepivacaine 2% with adrenaline. The mean maximal cephalad analgesic level was T9. At that abdominal segment, RIEMG showed a reduction of 50% and TURNS of 20%. Isometric force and RIEMG recorded simultaneously in the quadriceps muscles during epidural anaesthesia displayed a linear relationship with a correlation coefficient of 0.91. TURNS was insensitive to force variations above 60% of maximum voluntary contraction. During the regression phase, 90% of both the initial force and RIEMG value was noted 180 min after the epidural injection. It is concluded that recording of RIEMG is a good method for quantitative assessment of motor blockade during epidural anaesthesia.