Monopolar Cautery Use in Pediatric Cochlear Implant Users.

OBJECTIVES: To determine the incidence and impact of monopolar cautery use in a cohort of pediatric cochlear implant (CI) users. STUDY DESIGN: Case series from a retrospective chart review and a systematic review of the literature. SETTING: Tertiary academic referral center. METHODS: CI patient charts from 2012 to 2021 were reviewed from a single pediatric hospital system to determine if monopolar cautery was used during a subsequent surgical procedure. In addition, a systematic review of the literature was performed to identify additional, relevant patients. Postoperative CI function was the primary outcome measure. RESULTS: In total, 190 patients underwent a surgical procedure following cochlear implantation in a single pediatric hospital system. Fifteen patients (7.9%) and 17 distinct surgical procedures were identified in which monopolar cautery was used. Seven of these 17 cases (41.2%) involved the head and neck, and 10 were performed below the clavicles. No patients experienced a device failure or a decline in CI performance following surgery. A systematic review identified an additional 4 patients who underwent a surgery that used monopolar cautery following cochlear implantation, and no change in CI function was identified. CONCLUSIONS: The present study adds additional support to the notion that monopolar cautery does not necessarily injure CI functionality. While the most risk adverse strategy when planning a surgical procedure for a CI patient is to avoid monopolar cautery use altogether, the use of cautery should not immediately be associated with implant dysfunction.

[Mesenchymal stem cells and rheumatism. State of the art]. / Mesenchymale Stammzellen und Rheuma. "State of the art".

Mesenchymal stem cells (MSC) are multipotent. They are able to regenerate tissue damage and, in parallel, inhibit inflammation and fibrosis. As they are non-immunogenic, MSCs can be safely transplanted using autologous and allogeneic procedures: an option for refractory connective tissue diseases and osteoarthritis.

Effects of Fenoldopam in the Pediatric Population: Fluid Status, Serum Biomarkers, and Hemodynamics: A Systematic Review and Meta-Analysis.

Fluid overload is a frequent complication in children during critical illness. Fluid restriction and diuretic agents have been the mainstay therapies so far. Fenoldopam, a selective dopamine-1 receptor agonist, is a diuretic agent with promising effects in the pediatric population. The purpose of this meta-analysis is to evaluate the outcomes of pediatric patients who received fenoldopam. We hypothesized that the administration of fenoldopam will cause an increase in urine output and decrease in serum creatinine in this patient population. A comprehensive database search of PubMed, EMBASE, and Cochrane libraries from the databases' inception through December 2018 was undertaken. Independent reviewers selected appropriate studies and the reviewed data. A meta-analysis was then conducted to determine the effects of fenoldopam on hemodynamics, the amount of vasoactive support, and renal function in children under the critical care setting. The selected end points were measured prior to the administration of fenoldopam and 24 hours after the initiation of the infusion: urine output, serum creatinine, serum sodium, inotrope score, heart rate, central venous pressure, systolic blood pressure, and mean blood pressure. Forest plots were generated to demonstrate individual study data as well as pooled data for each end point. A total of five studies (three retrospective cohort studies, two randomized trials) with 121 patients were included for analysis. No significant difference was observed in urine output, inotrope score, systolic blood pressure, or mean blood pressure. There was a statistically significant increase in serum creatinine and central venous pressure. There was statistically significant decrease in serum sodium and heart rate, and central venous pressure. This meta-analysis did not identify significant renoprotective or vasodilator effects from fenoldopam in this patient population. Although mild electrolyte and hemodynamic changes were identified, larger studies are warranted to determine the clinical significance of fenoldopam in this patient population.

[Treatment of radiation-induced late effects: What's new?] / Traitement des effets tardifs après la radiothérapie : quoi de neuf ?

Mechanisms of late radio-induced lesions are the result of multiple and complex phenomena, with many entangled cellular and tissue factors. The biological continuum between acute and late radio-induced effects will be described, with firstly a break in homeostasis that leads to cellular redistributions. New insights into late toxicity will finally be addressed. Individual radiosensitivity is a primary factor for the development of late toxicity, and clinicians urgently need predictive tests to offer truly personalized radiation therapy. An update will be made on the various functional and genetic tests currently being validated. The management of radio-induced side effects remains a frequent issue for radiation oncologists, and an update will be made for certain specific clinical situations. Finally, an innovative management for patients with significant side effects after pelvic radiotherapy will be developed, involved mesenchymal stem cell transplantation, with the presentation of the "PRISME" protocol currently open to patients recruitment.

Differential structural requirements for fibrinogen binding to platelets and to endothelial cells.

The cytoadhesins represent a group of RGD receptors that belongs to the integrin superfamily of adhesion molecules. Members of this cytoadhesin family include the platelet GPIIb-IIIa and the vitronectin receptors. These glycoproteins share the same beta-subunit, which is associated with different alpha subunits to form an alpha/beta heterodimer. In the present study, we have analyzed the fine recognition specificy of the cytoadhesins from platelets and endothelial cells for the adhesive protein, fibrinogen. Two sets of synthetic peptides, RGDX peptides and peptides corresponding to the COOH terminus of the fibrinogen gamma chain, were compared for their structure-function relationships in the two cellular systems. The results indicate that: (a) both RGDX and gamma-chain peptides inhibit the binding of fibrinogen to platelets and endothelial cells; (b) a marked influence of the residue at the COOH- and NH2-terminal positions of each peptide set can be demonstrated on the two types; and (c) RGDX and gamma peptides have differential effects on platelets and endothelial cells with respect to fine structural requirements. These results clearly indicate that while the platelet and endothelial cytoadhesins may interact with similar peptidic sequences, they express a different fine structural recognition.

The proteolytic action of bacterial proteases from Clostridium histolyticum on bovine fibrinogen.

Bacterial proteases from Clostridium histolyticum bring about the polymerization of fibrinogen into structured fibres, the period being 9 nm when the ionic strength (I) of the solution is between 0.1 and 0.2. At I=0.3 no polymerization occurs, but the successive actions of bacterial proteases and thrombin induce polymerization, the fibres obtained showing a periodic structure: the major period is 23 nm with two minor striations. The striation pattern is different from that obtained with fibrin which shows a major period of 23 nm with three minor striations. The degree of degradation of fibrinogen monitored by disc gel electrophoresis shows that the Aalpha polypeptide chain (mol. wt.=68 000) is degraded into Aalpha1 (mol. wt.=32 000) and Aalpha2 (mol. wt.=29 000), whilst the mobility of Bbeta increases and gamma remains unchanged; the molecular weight is estimated between 272 000 and 269 000. These results emphasize that the charge distribution differences which are compatible with the loss of different portions of the molecule, lead to variations in fibre structures.

Studies on factor VIII-related protein. I. Ultrastructural and electrophoretic heterogeneity of human factor VIII-related protein.

Human factor VIII-related protein precipitates with specific heterologous anti-bodies directed against purified factor VIII and supports ristocetin-induced aggregation of washed platelets. We purified human factor VIII from cryoprecipitate by subsequent gel filtration on crosslinked large-pore agarose. Factor VIII-related protein appeared as a large aggregate following electrophoresis on 3% polyacrylamide gels in the presence of sodium dodecyl sulfate (SDS). The same material was separated into multiple bands (molecular weight in excess of several millions) following electrophoresis on SDS-1% agarose gels. After complete disulfide reduction of factor VIII-related protein and electrophoresis on SDS-5% polyacrylamide gels a single subunit chain (Mr approximately equal to 200 000) was revealed. Analysis of this protein, in its non-reduced state, by negative contrast electron microscopy showed filaments of markedly variable size. The calculated molecular weight of such filaments ranged from about 0.6.10(6) to 20.10(6). We conclude that size heterogeneity is an essential feature of human factor VIII-related protein.

Transplantation of stromal cells transduced with the human IL3 gene to stimulate hematopoiesis in human fetal bone grafts in non-obese, diabetic-severe combined immunodeficiency mice.

The non-obese diabetic-severe combined immunodeficiency (NOD-SCID) mouse is a convenient host for human hematopoietic tissues and cells. Human fetal bone fragments engrafted subcutaneously in NOD-SCID mice sustain human hematopoiesis for several months. MS5 murine bone marrow stromal cells were transfected by electroporation with a plasmid containing the human interleukin-3 gene. As expected, stably transfected hu-IL3-MS5 cells supported human hematopoiesis in vitro more efficiently than MS5 cells. hu-IL3-MS5 cells were then injected intravenously into hu-NOD-SCID mice to test their ability to home to the mouse and/or human bone marrow, and to evaluate the role of hu-IL3 secretion on human hematopoiesis in vivo. hu-IL3 was detected in the mouse serum for up to an observation time of 8 weeks. hu-IL3-MS5 cells engrafted the bone marrow, spleen, liver and lungs of the mice but also the human bone graft. The presence of hu-IL3-MS5 cells in the human bone significantly stimulated local human hematopoiesis. This setting could be used to model the bone marrow homing of intravenously injected stromal cells or stromal cell precursors. The same experimental principle could also be applied in a therapeutic perspective to malignant human bone marrow hematopoiesis.

Monoclonal antibodies against human hemopoietic cells and the separation of progenitor cells from bone marrow.

Monoclonal antibodies against myeloid cell surface antigens were selected according to their reactivity against normal human bone marrow cells. Each of the antibodies detected a different population of maturing bone marrow cells, but almost none of the progenitor cells assayed (seven-day GM-CFC, 14-day GM-CFC, CFU-E, BFU-E). Myeloid cells, erythroid cells, and the majority of lymphocytes and monocytes were simultaneously depleted with the mixture of these three antibodies using fluorescence-activated cell sorting (FACS) or panning for cell separation; 14-day GM-CFC and BFU-E were enriched six- to 16-fold when negative cells were sorted. After a negative selection step using panning, BFU-E and 14-day GM-CFC were enriched four- to seven-fold and five- to 13-fold, respectively. Negative cells obtained after panning or cell sorting were also enriched in blast cells (24.4% and 27.2%, respectively) and depleted in maturing bone marrow cells, with the exception of variable numbers of lymphocytes, monocytes, and plasma cells. FACS of negative cells using both forward light-scatter and perpendicular light-scatter parameters resulted in a cell population that contained a majority of undifferentiated blasts and 5.5%-9.6% BFU-E and 14-day GM-CFC.

Targeted transfection of the IL-3 gene into primary human hematopoietic progenitor cells through the c-kit receptor.

We recently showed that an antibody-mediated gene transfer procedure termed antifection can be used for targeted gene delivery into lymphoid cells in vitro and in vivo. We here report that antifection also is effective for targeted gene transfer to immature hematopoietic cells. A human IL3-expressing plasmid was chemically linked to an anti-human CD117 antibody. Delivery of the IL3 plasmid into IL-3-dependent myeloid TF-1 cells (bearing the CD117 antigen) was specific and resulted in the transient proliferation of the targeted cells in the absence of exogenous IL-3. Transfection of primary human CD34+ hematopoietic stem/progenitor cells led to transient production of IL-3 and transient proliferation of the target cells. Interestingly, by using a semisolid progenitor cell assay, we found that transfected primary CD34+ cells were able to generate normal numbers of cell colonies in the absence of exogenous IL-3. Polymerase chain reaction analysis confirmed the presence and expression of the IL-3 transgene in the progenitor-derived colonies. In conclusion, our data show that CD117 is a suitable cell surface target to specifically transfer gene by antifection into primary CD34+ cells and that delivery of IL-3 gene in these cells resulted in the expression of a functional IL-3 able to support cell growth in absence of exogenous cytokine. Thus, antifection may provide new therapeutic modality relying on the transient production of appropriate growth factors acting via autocrine and/or paracrine mechanisms.

A synthetic peptide with anti-platelet activity derived from a CDR of an anti-GPIIb-IIIa antibody.

A monoclonal antibody, AC7, directed against the RGD (Arg-Gly-Asp) binding site on the GpIIIa subunit of the platelet fibrinogen receptor, interacts only with activated platelet. In order to identify the regions of AC7 that interact with the receptor, cDNA sequences of AC7 immunoglobulin heavy and light chain variable regions were determined. Among the six complementarity-determining regions (CDRs) of AC7, the CDR3 heavy chain (H3) contains homology to the RGDF sequence within fibrinogen. A synthetic peptide encompassing the H3 region (H3, RQMIRGYFDV) inhibited platelet aggregation and fibrinogen binding to platelet (IC50 = 700 microM). The inhibitory potencies of modified H3 peptides suggest that the RGYF sequence within the H3 peptide mimic the receptor recognition sequence in fibrinogen.

Immuno-purification of a dimeric subcomplex of the respiratory NADH-CoQ reductase of Rhodobacter capsulatus equivalent to the FP fraction of the mitochondrial complex I.

The Rhodobacter capsulatus genes encoding the NUOE and NUOF subunits, equivalent to the 24 kDa and 51 kDa subunits of the mammalian mitochondrial complex I, have been sequenced. According to the nucleotide sequence, the NUOE subunit is 389 amino acids long and has a molecular mass of 41.3 kDa. In comparison to the mitochondrial equivalent subunit, NUOE is extended at the C terminus by more than 150 amino acids. The NUOF subunit is 431 amino acids long and has a molecular mass of 47.1 kDa. A subcomplex containing both the NUOE and NUOF subunits was extracted by detergent treatment of R. capsulatus membranes and immuno-purified. This subcomplex is homologous to the mitochondrial FP fragment. Mass spectrometry after trypsin treatment of the NUOE subunit validates the atypical primary structure deduced from the sequence of the gene.

Antibody-mediated endocytosis of G250 tumor-associated antigen allows targeted gene transfer to human renal cell carcinoma in vitro.

Specific gene transfer into targeted tumor cells remains a critical issue for the development of systemic gene therapy protocols. With this end in view, we have tested the possibility of selectively directing genes to tumor cells through the recognition of tumor-associated antigens (TAA). This was approached in vitro on four human renal cell carcinoma (RCC) lines by means of the highly specific mouse G250 monoclonal antibody (mAb) chemically conjugated to a plasmid DNA conveying a reporter activity. This mAb directed to a TAA that is present on 95% of primary RCCs and on 60% of metastatic human RCCs was extensively characterized, including during clinical trials. Epifluorescence microscopy analysis indicated that upon specific binding to G250 TAA, G250 mAb alone or conjugated to plasmid DNA was internalized by an active endocytic process and colocalized with the transferrin concentrated in the late recycling perinuclear compartment. We also observed that both unconjugated G250 mAb or G250 mAb conjugated to plasmid DNA remained in the perinuclear region of the cells for > or = 20 hours and were not rapidly translocated to lysosomes or recycled to the plasma membrane. In contrast, unconjugated plasmid DNA was not internalized. After transfection of G250 TAA-positive RCC lines with G250 mAb conjugated to a plasmid cDNA encoding mouse interleukin-2, a significant and sustained production of mouse interleukin-2 protein was detected from days 5-15 and was abrogated by inhibiting the internalization process. Altogether, our data showed that endocytosis of G250 TAA should be the basis of gene transfer to RCC, suggesting that targeting of TAA capable of internalization may be the basis of new approaches for designing alternative cancer gene therapy procedures.

The fibrinolytic action of flufenamate and its influence on plasminogen binding to fibrin.

The effects of the synthetic fibrinolytic agent flufenamate were investigated in a purified system made up of bovine fibrin and human plasminogen. The lysis of the fibrin clots was observed after a 12-hour incubation for flufenamate concentrations ranging from 0.25 to 3 mM. Below 0.25 mM and above 3 mM, no lysis occurred, even after a longer incubation. An increase in plasminogen adsorption on the fibrin clot was also observed in the presence of flufenamate. The amount of plasminogen bound was estimated using 125I labelled Glu-plasminogen or Lys-plasminogen, in the presence of a protease inhibitor to avoid the fibrinolysis induced at lytic concentrations of flufenamate. Maximum binding was observed with both types of plasminogen at a flufenamate concentration of 2.5 mM. The binding, relatively fast at the start of incubation, slowed down progressively, but a real plateau was not reached, even after a 6-day incubation. Plasminogen binding was not saturable, suggesting a non-specific binding type. Although Lys-plasminogen binding was always greater than that of Glu-plasminogen, the effect of flufenamate was more pronounced on Glu-plasminogen binding.

Persistence of altered 5-hydroxytryptamine turnover following hemibody X-irradiation in the rat distal colon.

PURPOSE: Acute gastrointestinal responses to ionizing radiation exposure include a role for 5-hydroxytryptamine (5-HT), but it is not known whether involvement of 5-HT persists and contributes to late effects. The aim was to investigate the acute and later effects of lower hemibody irradiation on 5-HT turnover and the biological effect in the rat distal colon. MATERIALS AND METHODS: Rats were exposed to 10 Gy lower hemibody X-radiation. 5-HT and 5-hydroxyindole acetic acid tissue levels were measured in the distal colon along with the serotonin re-uptake transporter and tryptophan hydroxylase mRNA. 5-HT-containing cells and crypt cell numbers were estimated in addition to 5-HT-stimulated short-circuit current responses in isolated mucosa. Studies were performed from 3 days to 3 months post-exposure. RESULTS: During the acute phase, at 3 days post-irradiation, reductions in cell number, tissue resistance, serotonin re-uptake transporter expression and secretory responses to 5-HT were observed. However, at later times when secretory responses were normal, 5-HT tissue levels and enterochromaffin cell numbers were increased. CONCLUSIONS: The results provide evidence that after 10 Gy hemibody irradiation, modifications persist past the acute phase. In particular, 5-HT turnover in the distal colon is altered during a longer period.

Determination of residues of flumequine and 7-hydroxyflumequine in edible sheep tissues by liquid chromatography with fluorimetric and ultraviolet detection.

A simple, sensitive, and rapid method for simultaneous determination of residues of flumequine and its microbiologically active metabolite 7-hydroxyflumequine in 100 mg sheep edible tissues (muscle, liver, kidney, and fat) by liquid chromatography is reported. After liquid-liquid cleanup with ethyl acetate, tissue extracts were injected onto a Select B column. The 2 compounds were determined by ultraviolet and fluorimetric detection. The method was repeatable and reproducible for flumequine and 7-hydroxyflumequine in muscle, liver, kidney, and fat, with limits of detection below 2 and 3 micrograms/kg for flumequine and 7-hydroxyflumequine, respectively. Mean recoveries for flumequine were 90 +/- 7, 82 +/- 7, 89 +/- 5, and 82 +/- 6% in muscle, liver, kidney, and fat respectively. Mean recoveries for 7-hydroxyflumequine were 91 +/- 2, 90 +/- 4, 86 +/- 3, and 84 +/- 4% in muscle, liver, kidney, and fat, respectively.

[Domiciliary care of pathological pregnancies by midwives. Comparative controlled study on 996 women (author's transl)]. / Surveillance à domicile des grossesses pathologiques par les sages-femmes. Essai comparatif contrôlé sur 996 femmes.

In order to evaluate the effectiveness of domiciliary care of pathological pregnancies by midwives a comparative study was carried out on two groups of pregnant women, the one group being treated by this new method of caring fort them, the other by traditional care. There was random selection for the two groups. The results of this comparative controlled study show that when midwives care for these patients at home once a pathological condition has become established premature labour and admission to hospital is not avoided. A more preventive attitude developed. This implies that they intervene very early in pregnancy as soon as risk factors for pathological conditions appear without waiting for the pathological conditions themselves to develop.

Early lesions induced in rat colon epithelium by N-methyl-N'-nitro-N-nitrosoguanidine.

The development of ACF (aberrant crypt foci), adenoma and cancer following intrarectal administration of the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) has been described. However, microscopic lesions not previously reported were observed as soon as two weeks following carcinogen treatment. These lesions protrude slightly over the epithelial lining of the colon, with a micropolyp-like appearance. Oriented sections show that the centre of these lesions present pseudo-"cystic" appearance, with disorganized crypts made of normal cells. The chorion of the lesion is invaded by numerous inflammatory cells and some ACF may be present nearby. The epithelium lining the cysts and the distorted crypts shows expression of gastric mucin M1/MUC5AC, an early marker of colonic carcinogenesis which is not present in normal colon. This mucin is retained within the "cysts" together with some inflammatory cells. The micropolyps observed contain in a minute form some histological elements described in ulcerative colitis or short-term radiotherapy (distortion of crypts, crypt abscesses, increase of chorion cellularity, infiltration by immune cells). In addition, the presence of bifid crypts nearby suggests mucosal regeneration. Our hypothesis is that these modifications are steps in a normal healing pathway that may in some cases degenerate into precancerous lesions and cancer.