Few-mode erbium-doped fiber amplifier with photonic lantern for pump spatial mode control.

We demonstrate a few-mode erbium-doped fiber amplifier employing a mode-selective photonic lantern for controlling the modal content of the pump light. Amplification of six spatial modes in a 5 m long erbium-doped fiber to ∼6.2 dBm average power is obtained while maintaining high modal fidelity. Through mode-selective forward pumping of the two degenerate LP21 modes operating at 976 nm, differential modal gains of <1 dB between all modes and signal gains of ∼16 dB at 1550 nm are achieved. In addition, low differential modal gain for near-full C-band operation is demonstrated.

Clusters dissolution of Yb3+ in codoped SiO2-Al2O3-P2O5 glass fiber and its relevance to photodarkening.

Using a combination of pulse electron paramagnetic resonance and photoluminescence spectroscopy, we demonstrate the major role of phosphorous rather than aluminium in the rare-earth dissolution process, an essential advance in telecommunication and solid laser fields. Our results also provide new insight into the micro-structural origin of the photodarkening process occurring in Yb doped fiber.

Negative and translation termination-dependent positive control of FLI-1 protein synthesis by conserved overlapping 5' upstream open reading frames in Fli-1 mRNA.

The proto-oncogene Fli-1 encodes a transcription factor of the ets family whose overexpression is associated with multiple virally induced leukemias in mouse, inhibits murine and avian erythroid cell differentiation, and induces drastic perturbations of early development in Xenopus. This study demonstrates the surprisingly sophisticated regulation of Fli-1 mRNA translation. We establish that two FLI-1 protein isoforms (of 51 and 48 kDa) detected by Western blotting in vivo are synthesized by alternative translation initiation through the use of two highly conserved in-frame initiation codons, AUG +1 and AUG +100. Furthermore, we show that the synthesis of these two FLI-1 isoforms is regulated by two short overlapping 5' upstream open reading frames (uORF) beginning at two highly conserved upstream initiation codons, AUG -41 and GUG -37, and terminating at two highly conserved stop codons, UGA +35 and UAA +15. The mutational analysis of these two 5' uORF revealed that each of them negatively regulates FLI-1 protein synthesis by precluding cap-dependent scanning to the 48- and 51-kDa AUG codons. Simultaneously, the translation termination of the two 5' uORF appears to enhance 48-kDa protein synthesis, by allowing downstream reinitiation at the 48-kDa AUG codon, and 51-kDa protein synthesis, by allowing scanning ribosomes to pile up and consequently allowing upstream initiation at the 51-kDa AUG codon. To our knowledge, this is the first example of a cellular mRNA displaying overlapping 5' uORF whose translation termination appears to be involved in the positive control of translation initiation at both downstream and upstream initiation codons.

Spi-1/PU.1 is a positive regulator of the Fli-1 gene involved in inhibition of erythroid differentiation in friend erythroleukemic cell lines.

Spi-1/PU.1 and Fli-1 are two members of the ETS family of transcription factors whose expression is deregulated by proviral insertion in most erythroleukemic cell lines induced by the spleen focus-forming virus (SFFV) and Friend murine leukemia virus (F-MuLV) components of the Friend viral complex, respectively. In this study, we present evidence that transcription of the Fli-1 gene is positively regulated by Spi-1/PU.1 in SFFV-transformed cell lines: (i) all SFFV-transformed cell lines expressing Spi-1/PU.1 are characterized by a specific pattern of Fli-1 gene transcripts initiated in the -200 region instead of position -400 as reported for F-MuLV-transformed cell lines; (ii) these Fli-1 transcripts initiated in the -200 region are downregulated in parallel with that of Spi-1/PU.1 during hexamethylenebisacetamide (HMBA) induced differentiation; and (iii) Fli-1 transcription is upregulated in SFFV cells lines following stable transfection of a Spi-1/PU.1 expression vector. Furthermore, we found by transient transfection assays that the -270/-41 region of the Fli-1 gene displays promoter activity which is transactivated by Spi-1/PU.1. This promoter is strictly dependent on the integrity of two highly conserved ETS DNA binding sites that bind the Spi-1/PU.1 protein in vitro. Finally, we show that transfection of constitutive or inducible Fli-1 expression vectors in SFFV-transformed cells inhibits their erythroid differentiation induced by HMBA. Overall, these data indicate that Fli-1 is a target gene of the Spi-1/PU.1 transcription factor in SFFV-transformed cell lines. We further suggest that deregulated synthesis of Fli-1 may trigger a common mechanism contributing to erythroleukemia induced by either SFFV or F-MuLV.

Unexpected and coordinated expression of Spi-1, Fli-1, and megakaryocytic genes in four Epo-dependent cell lines established from transgenic mice displaying erythroid-specific expression of a thermosensitive SV40 T antigen.

Most erythroleukemic cell lines established in vitro coexpress erythrocytic and megakaryocytic markers that often are associated with expression of Spi-1 and/or Fli-1 transcription factors known as transactivators of megakaryocyte-specific promoters. In the present study, we examined the possibility of establishing new cell lines keeping strictly erythroid-specific properties in vitro through the targeted and conditional immortalization of erythrocytic progenitors. For that purpose, we established several lines of transgenic mice displaying erythroid-specific expression of a thermosensitive SV40 T antigen. As expected, these transgenic mice developed splenomegaly due to the massive amplification of Ter 119 positive erythroid nucleated cells expressing T antigen. Despite this drastic effect in vivo, the in vitro immortalization of erythropoietin-dependent erythroid progenitors unexpectedly occurred at low frequency, and all four cell lines established expressed both erythrocytic (globins) and megakaryocytic markers (glycoprotein IIb, platelet factor 4) as well as Spi-1 and Fli-1 transcripts at permissive temperature. Switching the cells to the nonpermissive temperature led to a marked increase in globin gene expression and concomitant decrease in expression of Spi-1, Fli-1, and megakaryocytic genes in an erythropoietin-dependent manner. Interestingly, enhanced expression of Spi-1 and Fli-1 genes already was detected in the Ter 119 positive cell population of transgenic mice spleen in vivo. However, like normal Ter 119 erythroid cells, these Ter 119 positive cells from transgenic mice still expressed high levels of beta-globin and very low or undetectable glycoprotein IIb and platelet factor 4 megakaryocytic transcripts. Taken together, these data indicate that the unexpected expression of megakaryocytic genes is a specific property of immortalized cells that cannot be explained only by enhanced expression of Spi-1 and/or Fli-1 genes.

Delta zero-thalassemia in cis of beta Knossos-globin gene. Normal structure transient expression of the delta-globin gene.

We have previously described the first homozygous cases of Hb Knossos in an Algerian family. The Hb A2 was completely absent, ascertaining the presence of a delta zero-thalassemia determinant in cis of the beta Knossos S gene. Here, we investigate the affected delta-globin gene. The complete DNA sequence of the gene and its 5' and 3' flanking regions was determined. Only two nucleotide changes were recorded: a C----T substitution at -199 and an AT insertion at -448 upstream from the cap site. To examine the involvement of these changes in gene function, the delta-gene was subcloned in an expression vector and introduced into COS cells. Analysis of RNA derived from these cells, using an S1 protection assay and dot-blot hybridization, revealed qualitatively and quantitatively normal transcription. The loss of delta-globin gene activity in vivo may be due to the alteration of a tissue-specific control.

[Accidents that can occur during fetal direct electro-cardiographic recording. Concerning a fracture of the scalp electrode (author's transl)]. / Les accidents de l'électrocardiographie foetale directe (à propos d'une fracture de l'électrode de scalp).

The authors report a case of fracture of a double spiral scalp electrode. One of the two spirals stayed in the scalp tissue when the electrode was being removed. This spiral was localised by X-Rays and was removed under local anaesthetic. There were no complications. No similar case of such an accident has been reported in the literature and this is the first happening in our 2,472 cases of monitoring. Of the other serious complications which are mentioned in the literature, abscess of the scalp is the most frequent at 0.4 per cent (9). We have had no complications in 2,472 cases of monitoring. The outcome of such an abscess is usually quite benign, but there have been two cases of septicaemia followed by death reported. The complication of infection is serious and has to be treated carefully by disinfection. Two other complications have been noted and that is, tearing or ripping off a little of the scalp (2 cases in 2,472 monitorings). Spinal fluid leakage has been reported when an electrode was placed on a fontanelle and haemorrhage. Most of these complications took place when the clip electrodes which have now been abandoned were used and, their frequency being rare, does not invalidate the definite advantages of direct monitoring of the fetus in labour.

[Prolapse after hysterectomy. A study of 45 cases (author's transl)]. / Les prolapsus aprés hystérectomie. A propos d'une série de 45 observations.

The authors report 45 cases of prolapse occurring after hysterectomy (26 after subtotal hysterectomy, 9 after total abdominal hysterectomy and 10 after total vaginal hysterectomy). These prolapses are rare and their incidence does not seem to vary with the type of hysterectomy that preceded them. although in some cases hysterectomy could be incriminated as the cause of the prolapse, in the majority of cases the reason was a prolapse that had been neglected when the hysterectomy had been carried out, or a prolapse that appeared a long time after hysterectomy because of the inevitable ageing of the supporting tissues of the pelvis. From the anatomical point of view it is important to distinguish those prolapses where the vaginal vault does not descend and those where there is total descent including the vault of the vagina. The prolapses give rise to difficult problems of therapy. The choice of operation has to take into account anatomical components of the prolapse, the functional repercussions, the urinary symptoms and whether the patient wishes to does not wish to continue sexual activity. If it is not necessary to keep the vagina open an operation that involves colpectomy or colpocervicectomy can give rise to very good anatomical and urinary results. When it is necessary to keep the vagina functioning as a vagina in the case of prolapse after subtotal hysterectomy, it is important to treat the case as though on was dealing with an ordinary prolapse. All the same, when dealing with procidentia it may be wiser to add a colpopexy procedure by the abdominal route. When dealing with a prolapse after total hysterectomy when the vaginal vault is in place, it is sufficient to carry out the usual form of perineal plastic operation general;y to obtain a good result, but when the vaginal vault has come down it is as well to carry out a colpopexy procedure by the abdominal route.

[Grave obstetrical phlebitis. Physiopathology, diagnosis and treatment (6 case histories) (author's transl)]. / Phlébites obstétricales graves. Physiopathologie, diagnostic et traitement (6 observations).

The authors detail their concepts of the physiopathology, the diagnostic methods and the treatments of grave puerperal phlebitis, having seen six cases in their two departments recently. They consist of 5 cases of iliofemoral thrombosis, 2 of whom were diagnosed during their pregnancies and 3 others whom were diagnosed after delivery. One of these died of pulmonary embolus: and there was one case of thrombosis of the right ovarian vein during post-partum. Over and above the classical factors that predispose to this condition in pregnancy, the authors draw attention to the anatomical constitutional factors that have been observed by Cockett in the physiopathology of these cases. The diagnosis is made using non-invasive methods: Doppler, plethysmography, labelled fibrinogen and isotope phlebography after delivery, supplemented when the results are positive by radiography of the iliac and vena caval systems which alone gives a precise diagnosis of the site. Therapeutic possibilities change according to the time that the condition is perceived, according to the topography of the lesions, and according to the existence or non-existence of moving thrombi. The treatment is directed to avoiding the complications of emboli and to preventing secondary functional sequellae. Finally the gynaecological problems of contraception and of further pregnancies are considered.

Liquid-chromatographic measurement of purine nucleotides in blood cells.

In this anion-exchange "high-performance" liquid-chromatographic method of analysis for purine nucleotides, the nucleotides are separated with high efficiency and selectivity on a weak anion exchanger (Hypersil APS 2, 3-micron particle size) by elution with a gradient of eluent pH and concentration. Applying this method to analysis for these compounds in human blood cells, we determined them in a patient with adenosine deaminase deficiency who was treated with a bone-marrow transplantation, finding that the transplantation did not entirely correct the patient's abnormalities of purine metabolism.

[Separation of herbicides by high-performance liquid chromatography. Influence of water (author's transl)]. / Séparation d'herbicides par chromatographie en phase liquide à haute performance. Influence de l'eau

The influence of the amount of water dissolved in dichloromethane on the chromatographic separation of herbicides was studied. The selectivity of the mobile phase was demonstrated and compared to another system, dichloromethane modified with 2-propanol. The high efficiency of microporous packing was also demonstrated.

[Separation of pharmaceutical compounds using high-performance liquid chromatography. Influence of water II. (author's transl)]. / Séparation de produits pharmaceutiques par chromatographie en phase liquide à haute performance. Influence de l'eau. II

The use of ternary solvent systems as mobile phases in combination with highly efficient chromatographic columns is of particular interest for the analysis of pharmaceutical and biological compounds. The solute-silanol group interactions decrease when the eluent is blended with water. Good selectivities are found with such systems.

Hypoxanthine and xanthine levels determined by high-performance liquid chromatography in plasma, erythrocyte, and urine samples from healthy subjects: the problem of hypoxanthine level evolution as a function of time.

The levels of hypoxanthine and xanthine are determined in plasma, erythrocyte, and urine samples by a reverse-phase high-performance liquid chromatographic (HPLC) method. The hypoxanthine concentration increases in erythrocyte and plasma samples when whole blood is stored at room temperature between sampling and centrifugation. Furthermore, the hypoxanthine concentration increases in erythrocyte samples when they are kept apart at room temperature before analysis, whereas the plasma hypoxanthine level remains constant. This result proves an endogenous formation of hypoxanthine in erythrocytes with time, at room temperature. These studies show the necessity of rigorous conditions for the collection, transport, and treatment of blood samples. In order to achieve accurate results, the blood must be centrifuged immediately after collection. The erythrocyte and plasma samples must be stored frozen until deproteinization and HPLC analysis. Under these conditions, the concentrations of hypoxanthine and xanthine in plasma are 2.5 +/- 1 and 1.4 +/- 0.7 microM, respectively. In erythrocyte samples, hypoxanthine concentration reaches 8.0 +/- 6.2 microM.

High-performance liquid chromatographic determination of hypoxanthine and xanthine in biological fluids.

A rapid and selective reversed-phase high-performance liquid chromatographic method for the simultaneous determination of hypoxanthine and xanthine in biological fluids was developed. The identification of hypoxanthine and xanthine was confirmed by xanthine oxidase reaction. This method was applied to the investigation of purine metabolism in subjects with xanthine oxidase deficiency or gout. Hypoxanthine concentrations three to ten times higher than those determined in plasma were found in erythrocyte samples from normal subjects and from patients with xanthine oxidase deficiency or hyperuricemia under allopurinol therapy.

Ion-pair extraction and high-performance liquid chromatographic determination of tianeptine and its metabolites in human plasma, urine and tissues.

An isocratic reversed-phase ion-pair liquid chromatographic method for the determination of tianeptine and its two main metabolites in plasma, urine and tissues, using an internal standard, is reported. The influence of two stationary phases on the retention of the drugs was studied. The drugs were extracted as ion pairs, using a heptane-octanol-tetraheptylammonium bromide mixture (98:2:0.5, v/v/w) as extraction solvent. This extraction procedure yielded plasma drug recoveries of greater than 60% and allowed UV detection at 220 nm without interference from endogenous components of plasma, urine or tissues. Linear standard curves up to 1.00 micrograms/ml and drug determination down to 0.01 microgram/ml were observed. This method has been successfully applied to the analysis of human plasma and urine samples and of encephales from tianeptine-dosed rats.

Rapid assay of N-acetyl-beta-D-glucosaminidase isoenzymes in urine by ion-exchange chromatography.

We describe a new method for separating and measuring urinary N-acetyl-beta-D-glucosaminidase isoenzymes by "high-performance" liquid chromatography. Isoenzymes are eluted from the anion-exchange column with a one-step linear gradient of NaCl solution. For continuous post-column quantification of the activities of these isoenzymes, we use an on-line post-column reactor and 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide as substrate; the methylumbelliferone formed in this reaction is quantified fluorimetrically. We discuss the effects of varying different components of the assay: NaCl concentration, the pH of the mobile phase and of the reaction reagent, substrate concentration, incubation temperature, and the geometry of the post-column reactor.

Beta+-thalassemia in cis of a sickle cell gene: occurrence of a promoter mutation on a beta s chromosome.

An atypical sickle cell trait with a very low level of hemoglobin S and features of heterozygous beta-thalassemia was recently described. In vitro globin chain synthesis strongly suggested the presence of the two abnormalities on the same chromosome. We report the corresponding beta S-thal gene. DNA sequence revealed a C----T base substitution in the distal promoter element CACCC, at position-88 from the cap site, in addition to the expected GAG----GTG mutation responsible for the structural variant (beta 6 Glu----Val). Reticulocyte mRNA titration and transient assay of the mutant gene in COS cells showed a defect in beta-mRNA production. Restriction haplotype and DNA sequence analyses revealed that the doubly mutated gene is associated with haplotype 19 (or Benin/Algeria haplotype). In particular, we found the (AT)9(T)4 repeated sequences specifically encountered 5' to the beta S gene of Benin Algeria type. These results support the view that the beta S-thal gene resulted from an independent thalassemic mutation having occurred on a beta S chromosome rather than (a) from a beta S mutation having altered a beta-thalassemic gene or (b) from a recombination event between two chromosomes, each carrying one of the mutations.

Structural characterization of the beta-globin gene cluster in an individual expressing a very low level of G gamma globin chains.

We report the case of a normal individual displaying an extremely unbalanced G gamma/A gamma-globin ratio (G gamma-globin chains undetectable by urea/triton/ acrylamide gel electrophoresis and just reaching the threshold of detection by high performance liquid chromatography) associated with a very low level of G gamma-globin mRNA (at the most 5% of total gamma-mRNA after reverse transcriptase polymerase chain reaction determination). By DNA Southern blotting and sequencing, the very low level of G gamma-globin chains in this individual was found in association with subhaplotype [+ -----] (Hinc II 5' to epsilon, Xmn I 5' to G gamma, Hind III in G gamma and A gamma, Hinc II in and 3' to psi beta), with G gamma- and A gamma-globin gene sequences of the B type chromosome, and with a number of AT repeats in the locus control region hypersensitive site-2 site, similar to that reported to be associated with the Bantu beta S haplotype. These structural characteristics, described for the first time combined in the same individual, suggest that the G gamma/A gamma ratio in adults, is controlled by sequences distributed all along the beta-globin gene cluster.

Simultaneous plasma determination of floctafenin and its major metabolites by high-performance liquid chromatography: preliminary observations in children.

An isocratic reversed-phase ion-pair liquid chromatography with UV detection at 350 nm for the determination in human plasma of floctafenin (F) and its three main metabolites--floctafenic acid (FA), hydroxyfloctafenin (HOF), and hydroxyfloctafenic acid (HOFA)--is reported. Analytes and internal standard were extracted from acid plasma into ethyl acetate, and this organic phase was evaporated to dryness. This extraction yielded plasma drug recoveries of greater than 72%. Using 1 ml of plasma, the lower quantification limit was 0.05 microgram ml-1 with excellent linearity up to 0.8 microgram ml-1 for HOF and HOFA and up to 4.0 micrograms ml-1 for F and FA. The reproducibility and the selectivity of the method for several drugs thought likely to be administered in conjunction with F, were demonstrated. This method has been successfully applied to a pharmacokinetic study with a single 10 mg kg-1 oral dose in ten children.

A new alpha chain variant Hb Sallanches [alpha 2 104(G11) Cys-->Tyr] associated with HbH disease in one homozygous patient.

We identified a new alpha-chain variant (alpha Sal) associated with haemolytic anaemia and low level of HbH in one homozygous patient. This new mutation is located in codon 104 (TGC-->TAC) of the alpha 2 globin gene and results in a Cys-->Tyr replacement. In vitro and in vivo biosynthetic studies suggest that the mechanism leading to HbH disease in this homozygous patient is mostly related to a significant instability of alpha Sal:beta dimers rather than to the hyperinstability of the alpha Sal chain itself only.