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  • 1
    Keywords: RECEPTOR ; CELLS ; EXPRESSION ; CELL ; GENERATION ; screening ; POPULATION ; SITE ; PROTEIN ; cell line ; MOLECULES ; LINES ; RELEASE ; INDUCTION ; CELL-LINES ; DOWN-REGULATION ; treatment ; MOLECULE ; culture ; GLYCOPROTEIN ; PARTICLES ; TARGET ; virus ; MUTANT ; ASSAY ; MUTATION ; CELL-LINE ; LINE ; VIRUS-LIKE PARTICLES ; LIPID RAFTS ; LIVING CELLS ; RECEPTORS ; FLUORESCENCE ; cell lines ; HIGH-LEVEL ; HUMAN-IMMUNODEFICIENCY-VIRUS ; CYTOTOXICITY ; PRODUCTS ; VIRIONS ; LEVEL ; RECOVERY ; VIRAL INFECTIVITY ; antiviral ; HISTONE ACETYLATION ; COMPOUND ; packaging ; VARIETIES ; INCREASES ; HIV-1 NEF ; TYPE-1 NEF PROTEIN
    Abstract: Background: The availability of cell lines releasing fluorescent viral particles can significantly support a variety of investigations, including the study of virus-cell interaction and the screening of antiviral compounds. Regarding HIV-1, the recovery of such biologic reagents represents a very hard challenge due to the intrinsic cytotoxicity of many HIV-1 products. We sought to overcome such a limitation by using a cell line releasing HIV-1 particles in an inducible way, and by exploiting the ability of a HIV-1 Nef mutant to be incorporated in virions at quite high levels. Results: Here, we report the isolation and characterization of a HIV-1 packaging cell line, termed 18-4s, able to release valuable amounts of fluorescent HIV-1 based Virus-Like Particles (VLPs) in an inducible way. 18-4s cells were recovered by constitutively expressing the HIV-1 NefG3C mutant fused with the enhanced-green fluorescent protein (NefG3C-GFP) in a previously isolated inducible HIV-1 packaging cell line. The G3C mutation creates a palmitoylation site which results in NefG3C-GFP incorporation into virions greatly exceeding that of the wild type counterpart. Upon induction of 18-4s cells with ponasterone A and sodium butyrate, up to 4 mu g/ml of VLPs, which had incorporated about 150 molecules of NefG3C-GFP per viral particle, were released into the culture supernatant. Due to their intrinsic strong fluorescence, the 18-4s VLPs were easily detectable by a novel cytofluorometric-based assay developed here. The treatment of target cells with fluorescent 18-4 VLPs pseudotyped with different glycoprotein receptors resulted in these becoming fluorescent as early as two hours post-challenge. Conclusion: We created a stable cell line releasing fluorescent HIV-1 based VLPs upon induction useful for several applications including the study of virus-cell interactions and the screening of antiviral compounds
    Type of Publication: Journal article published
    PubMed ID: 17192195
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  • 2
    Keywords: RECEPTOR ; IN-VITRO ; CELL ; KINASE ; PATHWAY ; VITRO ; NEW-YORK ; GENE ; PROTEIN ; PROTEINS ; transcription ; DIFFERENTIATION ; ACTIVATION ; T cell ; T-CELL ; BINDING ; PHOSPHORYLATION ; ASSOCIATION ; ACID ; TARGET ; ELEMENT ; MUTANT ; MUTATION ; SIGNAL-TRANSDUCTION ; FRANCE ; RECRUITMENT ; leukocyte ; TCR ; TYROSINE PHOSPHORYLATION ; INTERLEUKIN-2 ; signaling ; INTERFERENCE ; T-CELL-ACTIVATION ; interaction ; GENE-TRANSCRIPTION ; TYROSINE KINASES ; ANTIGEN RECEPTORS ; progenitor ; USA ; UNIT ; immunology ; KINASE-1 ; interleukin ; MEDICINE ; ADAPTER PROTEIN ; LAT ; T-CELL-RECEPTOR ; ZAP-70
    Abstract: The SH2 domain-containing leukocyte protein of 76 kD (SLP-76) is a pivotal element of the signaling machinery controlling T cell receptor (TCR)-mediated activation. Here, we identify 14-3-3 epsilon and.. proteins as SLP-76 binding partners. This interaction was induced by TCR ligation and required phosphorylation of SLP-76 at serine 376. Ribonucleic acid interference and in vitro phosphorylation experiments showed that serine 376 is the target of the hematopoietic progenitor kinase 1 (HPK-1). Interestingly, either S376A mutation or HPK-1 knockdown resulted in increased TCR-induced tyrosine phosphorylation of SLP-76 and phospholipase C-gamma 1. Moreover, an SLP-76-S376A mutant induced higher interleukin 2 gene transcription than wild-type SLP-76. These data reveal a novel negative feedback loop involving HPK-1-dependent serine phosphorylation of SLP-76 and 14-3-3 protein recruitment, which tunes T cell activation
    Type of Publication: Journal article published
    PubMed ID: 17353368
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  • 3
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    German Medical Science; Düsseldorf, Köln
    In:  57. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC), Joint Meeting mit der Japanischen Gesellschaft für Neurochirurgie; 20060511-20060514; Essen; DOCP 10.152 /20060508/
    Publication Date: 2006-07-31
    Keywords: DSPA ; Intracerebral Hemorrhage ; Fibrinolysis ; Desmoteplase ; Intrazerebrale Blutung ; Fibrinolyse ; ddc: 610
    Language: English
    Type: conferenceObject
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  • 4
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    German Medical Science GMS Publishing House; Düsseldorf
    In:  Süddeutscher Kongress für Kinder- und Jugendmedizin; 64. Jahrestagung der Süddeutschen Gesellschaft für Kinder- und Jugendmedizin gemeinsam mit der Süddeutschen Gesellschaft für Kinderchirurgie und dem Berufsverband der Kinder- und Jugendärzte e.V. - Landesverband Bayern; 20150508-20150509; Nürnberg; DOC15sgkjP13 /20150325/
    Publication Date: 2015-03-26
    Keywords: ddc: 610
    Language: German
    Type: conferenceObject
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  • 5
    ISSN: 0378-1119
    Keywords: Recombinant DNA ; Tat transactivation ; long terminal repeat ; reporter gene ; transient expression ; virus titration
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1600-0595
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract – Based on an analysis of the literature concerning parameters influencing the prognosis of traumatic dental injuries, few studies were found to have examined possible relationships between treatment delay and pulpal and periodontal ligament healing complications. It has been commonly accepted that all injuries should be treated on an emergency basis, for the comfort of the patient and also to reduce wound healing complications. For practical and especially economic reasons, various approaches can be selected to fulfill such a demand, such as acute treatment (i.e. within a few hours), subacute (i.e. within the first 24 h), and delayed (i.e. after the first 24 h). In this survey the consequences of treatment delay on pulpal and periodontal healing have been analyzed for the various dental trauma groups. Applying such a treatment approach to the various types of injuries, the following treatment guidelines can be recommended, based on our present rather limited knowledge of the effect of treatment delay upon wound healing. Crown and crown/root fractures: Subacute or delayed approach. Root fractures: Acute or subacute approach. Alveolar fractures: Acute approach (evidence however questionable). Concussion and subluxation: Subacute approach. Extrusion and lateral luxation: Acute or subacute approach (evidence however questionable). Intrusion: Subacute approach (evidence however questionable). Avulsion: If the tooth is not replanted at the time of injury, acute approach; otherwise subacute. Primary tooth injury: Subacute approach, unless the primary tooth is displaced into the follicle of the permanent tooth or occlusal problems are present; in the latter instances, an acute approach should be chosen. These treatment guidelines are based on very limited evidence from the literature and should be revised as soon as more evidence about the effect of treatment delay becomes available.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1600-0595
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract  – Root resorption is known to be the most relevant complication determining the long-term prognosis of allotransplanted teeth, and it is initiated during the first few postoperative months. The aim of the present study was to quantitatively assess the dynamics of the periodontal ligament (PDL) and pulpal healing reactions during the first 8 weeks after allotransplantation of mature teeth. The material comprised 112 maxillary central and mandibular lateral incisors of 28 mature green Vervet monkeys, immunogenetically untested, and only matched according to the size of the grafts. Donors and recipients exchanged simultaneously both maxillary incisors and one mandibular incisor, whereas the contralateral mandibular incisors were autotransplanted as controls. At random, every second maxillary allograft was endodontically treated preoperatively. Histoquantitative analysis of the PDL and pulpal healing reactions was carried out after 1, 2, 4 and 8 weeks on serial cross-sections of the grafts in 6, 6, 6 and 8 monkeys, respectively. Necrosis zones in the PDL were prominent in both auto- and allografts after 1 week. Inflammation in the PDL dominated healing in all types of grafts 1 week after transplantation, whereas it subsided significantly after 2 weeks in autografts compared to allografts (P = 0.005). Inflammatory resorption (IR) became prominent after 4 weeks in autografts and this remained stationary. In contrast, IR initiated significantly earlier in allografts compared to autografts after 2 weeks (P = 0.007), and this type of resorption was further increasing in allografts after 4 and 8 weeks. Endodontic treatment, however, reduced IR nearly totally in the allografts with time. Replacement resorption (RR) was nearly absent in autografts. In contrast, allografts showed increasing appearance of RR with time, initiating at 4 weeks. By removing IR from the allografts by endodontic treatment, RR was unmasked significantly at 4 weeks (P = 0.02) and dominated most of the periodontal ligament (70%) after 8 weeks (P = 0.0004). Within the 8 postoperative weeks autografts showed healing with increasing amount of normal PDL reaching significantly higher levels compared to allografts already after 2 weeks (P = 0.02), with increasing differences thereafter. In most allografts, the normal PDL occupied less than 10% of the entire root surface and was located in the supra-alveolar cervical region. Downgrowth of periodontal pocket epithelium was absent or found very infrequently in all groups irrespective of type, time and treatment.In conclusion, the healing of allo- and autotransplanted mature teeth differed significantly on several aspects during the first 8 postoperative weeks. The recorded differences included a higher amount of inflammation in the PDL of allografts after 2 weeks, inflammatory resorption from the second week, and replacement resorption dominating in the eighth week, indicated that an immunologic stimulus for root resorption existed in the allogenic PDL apart from the pulp. Furthermore, specific healing reactions was found in the cervical region with almost identical gingival healing in auto- and allografts.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1600-0595
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract  – The effect of temperature of various storage media and at varying storage periods upon periodontal ligament (PDL) and pulpal healing after tooth replantation was examined in green vervet monkeys (Cercopithecus aethiops). Mandibular lateral incisors with mature root formation were extracted and kept in dry storage at 22, 4 and −18 °C; in saline at 37, 22, 4 and −18 °C; or in saliva (i.e. in the buccal vestibule) at 37 °C for either 60 or 120 min prior to replantation. The animals were sacrificed 8 weeks after replantation and the replanted teeth examined histometrically. The following histological parameters were registered for each tooth: normal PDL, surface resorption, inflammatory resorption, replacement resorption (ankylosis), downgrowth of pocket epithelium, periapical inflammatory changes, and the extent of vital pulp. A total of 125 replanted teeth were examined. Storage in saliva at 37 °C showed a similar amount of normal PDL compared to saline storage for both 60 and 120 min. Saline storage for 60 or 120 min showed no difference in the extent of normal PDL when storage was compared at 37, 22 and 4 °C. However, storage at −18 °C resulted in significantly less normal PDL than storage at other temperatures. Dry storage for 60 min showed significantly less root resorption at 4 °C compared to 22 °C. Furthermore, dry storage at −18 °C showed significantly less normal PDL than storage at 4 °C. When the dry storage period was extended to 120 min, no difference was found between 22, 4 and −18 °C. It is concluded that the temperature (above 0 °C) of the storage medium is of importance only for dry storage and in such a situation only for shorter extra-alveolar periods, i.e. for 60-min storage and not for 120 min, where extensive destruction of the PDL always takes place. It is suggested that the temperature effect of 4 °C could be related to less evaporation from the PDL and thereby less damage to the PDL cells or a strict temperature effect upon cell metabolism. Pulp healing in all the cases was limited to the entrance of the pulp canal, and no significant pattern was found between storage media, time and temperature.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Dental traumatology 15 (1999), S. 0 
    ISSN: 1600-0595
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract— The aim of the present study was to describe the frequency of dental injuries over a 10-year period, where a mouthguard was used in approximately 10% of intubation cases or endoscopying. The types of injury are presented and the use of different types of mouthguard is discussed. Among 120 086 procedures involving anaesthesia carried out in thc period 1983–1992 at Bispebjerg Hospital, Copenhagen, 75 cases of dental trauma occurred. The incidence of dental traumatic injuries was 0.06′%. Although eight patients had used mouthguards they were registered with traumatic injuries to the teeth in connection with general anaesthesia. Of the patients, 58% were women and 42% men, with a mean age of 61 years. Seventy-eight percent of the injuries were located in the maxilla and 91% of these involved the ineisor area of the maxilla. Forty-one percent were classified as lateral luxations or mobile teeth, 47% were classified as fractured teeth, 10% tooth avulsions and the remaining 2% were classified as concussions. Because of the low frequency of traumatic dental injuries in connection with general anaesthesia and the non-significant effect of the use of mouthguards, routine use of mouthguards is not recommended. Only when a patient is to have an endoscopy done under general anaesthesia, and the surgeon or anaesthesiologist foresees problems (restricted mouth opening, extensive fixed prosthetics, extensive loss of marginal bone) is use of an individually made mouthguard indicated.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    ISSN: 1600-0714
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Using histochemical and immunohistochemical methods, we examined specimens of Candidal leukoplakia from the oral mucosa of 5 smokers to determine the morphological relationships between Candida and Langerhans cells (LC) in tissue sections. LC were fairly evenly distributed in control sections, but had a patchy distribution in lesions. Fewer LC were found in lesions than in control tissue, but the difference was not statistically significant. Candidal antigens were not detected on LC by the methods used, but we found ATPase-positive LC among, or at least near, intraepithelial candidal hyphae. However, sections double-reacted with anti-Candida and T6 antibodies to label candidal antigens and LC. respectively, showed a clear zone of epithelium between the T6-positivc LC and the candidal hyphae. The difference in distribution of ATPase-positive and T6-positive LC may indicate locations of 2 subtypes of LC, or a change in T6 antigen expression by the LC closest to the candidal hyphae.
    Type of Medium: Electronic Resource
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