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Supplementary Tables 1-3, Figures 1-8 from Common Botanical Compounds Inhibit the Hedgehog Signaling Pathway in Prostate Cancer

Objectifs Le traitement de l’arteriopathie obliterante des membres inferieurs (AOMI) par technique endovasculaire en ambulatoire (HA) est une alternative a l’hospitalisation conventionnelle (HC). Nous rapportons dans ce travail les resultats cliniques en termes de securite et d’efficacite a 30 jours de la comparaison de ces deux prises en charge. Materiels AMBUVASC est une etude prospective, multicentrique controlee et randomisee. Les principaux criteres d’inclusion etaient l’egibilite a l’hospitalisation ambulatoire, l’ischemie d’effort, l’utilisation d’un introducteur de 5 a 7 French inclus, une ponction femorale retrograde. Les ponctions femorales retrogrades, radiales et humerales etaient exclues. Un systeme de fermeture arterielle etait utilise systematiquement dans le groupe HA et selon l’operateur dans le groupe HC. Le critere principal etait le ratio cout-utilite incremental. Les criteres secondaires etaient le deces, les complications ; les reinterventions et d’efficacite (rutherford, IPS). Resultats Cent-soixante patients etaient randomises (80 par groupe). Dans le bras HA, 4 patients changeaient de bras (refus de l’ambulatoire ; contre-indication de l’anesthesiste). En intention de traiter modifiee, 76 etaient analyses dans le bras HA et 77 dans le bras HC. Cent pour cent des patients etaient ASA 1, 2 ou 3 stable ; 95 % des patients etaient claudicants. Un stenting etait realise dans 96 % des cas dans le groupe HA et 86 % des cas dans le groupe HC (p = 0,03). Le succes technique etait de 96 % dans le groupe HA et de 99 % dans le groupe HC (p = 0,37). Quatre-vingt-neuf pour cent des patients en ambulatoire sortaient le soir meme. A 30 jours, on notait un deces dans le groupe HA non lie a l’AOMI. A 30 jours, 3 et 1 reinterventions etaient respectivement realisees dans les groupes HA et HC. Trois reinterventions etaient realisees en raison de complications au niveau de l’artere traitee et 1 complication en raison d’une thrombose au point de ponction. Aucune reintervention pour hemorragie n’etait realisee. Le taux de complications etait similaire entre le bras HA et HC (20 % vs 18 %, p = 0,81). Il n’y avait pas de difference entre les 2 groupes en termes d’amelioration clinique (p = 0,39) et hemodynamique (p = 0,84). Conclusion Le traitement de l’arteriopathie obliterante des membres inferieurs par technique endovasculaire en ambulatoire est sur et efficace.

Hintergrund Offene Kalkaneusfrakturen sind seltene schwere Verletzungen, die aufgrund der moglichen Komplikationen ein chirurgisch erfahrenes und interdisziplinares Team erfordern. Wie allgemein bei offenen Frakturen bestimmt der Grad der Weichteilverletzung die Therapie der Fraktur.

Gram-positive bacterial bone infections are an important cause of morbidity particularly in immunocompromised patients. Antimicrobial peptides (AP) are effectors of the innate immune system and directly kill microorganisms in the first hours after microbial infection. The aim of the present investigation was to study the expression and regulation of gram-positive specialized human beta-defensin-3 (HBD-3) in bone. Samples of healthy and osteomyelitic human bone were assessed for the expression of HBD-3. Using primary and immortalized osteoblasts (SAOS-2 cells), release and regulation of HBD-3 was evaluated after exposure to Staphylococcus aureus supernatant and/or corticosteroids using PCR, immunohistochemistry, Western blot and ELISA. To determine the role of toll-like-receptors-2 and -4 (TLR-2/-4), shRNA was used to downregulate TLRs. An osteomyelitis mouse model was created performed to investigate the release of murine beta-defensins using immunohistochemistry and RT-PCR. Cultured osteoblasts and human bone produce HBD-3 under standard conditions. The release increases within hours of bacterial supernatant exposure in cultured osteoblasts. This observation was not made in chronically infected bone samples. The shRNA-technology revealed the necessity of TLR-2 and -4 in HBD-3 induction in osteoblasts. Blocking protein synthesis with cycloheximide showed that the rapid release of HBD-3 is not dependent on a translational de novo synthesis and is not affected by glucocorticoids. The murine osteomyelitis model confirmed the in vivo release uptake of mouse beta-defensins-4 (MBD-4) in bone. This report shows the bacterial induction of HBD-3 via TLR-2 and -4 in osteoblasts and suggests a central role of antimicrobial peptides in the prevention of bacterial bone infection. The rapid and effective induction of HBD-3 in osteoblasts incubated with conditioned media from bacteria is more likely a result of a rapid secretion of preformed HBD-3 by osteoblasts rather than a result of enhanced biosynthesis. The increased incidence of gram-positive bacterial bone infection in patients with regular intake of glucocorticoids does not seem to be caused by a deranged HBD-3 release in osteoblasts.

Ziel der operativen Therapie pertrochantarer Femurfrakturen ist es, den praoperativen Mobilitatsgrad des Patienten schnellstmoglich wiederherzustellen. Es sollte untersucht werden, ob mit dem „proximal femoral nail“ (PFN) oder dem „trochanteric gamma nail“ (TGN) ein besseres operatives Ergebnis erzielt werden kann. Im Fokus standen operationstechnische Unterschiede und Fruhkomplikationen. In diese prospektive Studie konnten 114 der mit einem PFN und TGN versorgten Patienten eingeschlossen werden. Das Durchschnittsalter lag bei 78,9 Jahren. Es erfolgte eine klinische und radiologische Evaluierung. Bei isolierter Betrachtung der 31A1-Frakturen ergab sich eine um 20 min kurzere Operationsdauer beim proximalen Femurnagel (PFN). 78,5% aller Operationen verliefen komplikationslos. In 10 Fallen (7-mal PFN, 3-mal TGN) traten Probleme beim Einbringen des Nagels in den Markraum auf. Bei 12 (10,5%) Osteosynthesen [8-mal PFN (7%), 4-mal TGN (3,5%)] kam es postoperativ zu einer Dislokation des eingebrachten Osteosynthesematerials. Ein Cut-out trat beim PFN in 4 Fallen auf, einmal war der Z-Effekt die Ursache; 2-mal kam es zum Abkippen der Fraktur im Varussinn. In der TGN-Gruppe trat ein Cut-out in 2 Fallen auf. In einem Fall kam es zu einer sekundaren Varisierung ohne Cut-out. Ein signifikant hoheres Auftreten einer Komplikation lag bei keinem der verwendeten Implantate vor (p>0,05). Der PFN eignet sich aufgrund der hoheren Rotationsstabilitat des Doppelschraubensystems, besser zur Versorgung der 31A1-Frakturen. Bei geringer Femurschaftlange und bei starker Antekurvation des Femurs kommt es aufgrund des Nageldesigns zu Problemen bei der Insertion des PFN in den Markraum. Es ist sinnvoll, bereits praoperativ in Abhangigkeit von Frakturmorphologie und Patientenanatomie gezielt eines der beiden Implantate auszuwahlen.

Introduction Malalignment after osteosynthetic stabilization of lower leg fractures is still a common problem for trauma surgeons. The aim of the present study was to evaluate the incidence of torsional and varus- or valgus-malalignment of the lower leg subsequent to osteosynthetic stabilization techniques such as reamed nailing, unreamed nailing and tibial plating. Methods 70 patients with 73 fractures of the lower leg were included in the study. The fractures were treated consecutively in 37 cases with an unreamed nail (UTN), in 21 cases with a reamed nail and 15 cases were stabilized with a plate. During clinical follow-up after 5.7 years each patient was analyzed for malalignment of the lower leg with a CT-Scan and a dual-energy X-ray absorptiometry (DXA) analysis. Results Multi-level CT-scans revealed a significant rotational malalignment in 16.4 % of patients. Interestingly, all misaligned cases were treated with a nail (9.6 % UTN, 6.8 % reamed nail). Varus- or valgus-malalignment was detected in 5.4 % of cases all of whom had been treated with an intramedullary nail. Conclusions Malalignment is still a common problem after osteosynthetic stabilization of lower leg fractures, whereby the majority of these cases can be expected after intramedullary nailing. Rotational malalignment can be detected by CT-Scans, whereas DXA analysis is a reliable procedure to diagnose varus- or valgus-malalignment after osteosynthetic stabilization of lower leg fractures.

Sutherlandia frutescens is a medicinal plant, traditionally used to treat various types of human diseases, including cancer. Previous studies of several botanicals link suppression of prostate cancer growth with inhibition of the Gli/hedgehog (Gli/Hh) signaling pathway. Here we hypothesized the anti-cancer effect of S. frutescens was linked to its inhibition of the Gli/Hh signaling in prostate cancer. We found a dose- and time-dependent growth inhibition in human prostate cancer cells, PC3 and LNCaP, and mouse prostate cancer cell, TRAMP-C2, treated with S. frutescens methanol extract (SLE). We also observed a dose-dependent inhibition of the Gli-reporter activity in Shh Light II and TRAMP-C2QGli cells treated with SLE. In addition, SLE can inhibit Gli/Hh signaling by blocking Gli1 and Ptched1 gene expression in the presence of a Gli/Hh signaling agonist (SAG). A diet supplemented with S. frutescens suppressed the formation of poorly differentiated carcinoma in prostates of TRAMP mice. Finally, we found Sutherlandioside D was the most potent compound in the crude extract that could suppress Gli-reporter in Shh Light II cells. Together, this suggests that the S. frutescens extract may exert anti-cancer effect by targeting Gli/Hh signaling, and Sutherlandioside D is one of the active compounds.

Defensins are antibiotic peptides that are involved in host defence at epithelial and mesenchymal surfaces. Previous studies have shown the induction of human beta-defensin-3 (HBD-3) in osteoarthritic joints, suggesting that these molecules have functions in addition to their ability to kill microbes. The aim of this study was to investigate the production of a further human beta-defensin, named HBD-2, in osteoarthritis (OA) and to determine its regulation by inflammatory cytokines. Healthy and osteoarthritic cartilage was assessed for HBD-2 expression by RT-PCR, immunohistochemistry, and ELISA. C28/I2 chondrocytes, primary chondrocytes, and cartilage explants were cultured for in vitro studies. After 24 h of stimulation with tumour necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) or IL-6, real-time RT-PCR and ELISA experiments were performed to evaluate the effect of these cytokines on the production of HBD-2. In contrast to healthy cartilage, HBD-2 expression was identified in most of the OA samples examined (eight of ten). Cytokines that are potentially involved in the pathogenesis of OA, namely TNF-alpha, IL-1, and IL-6, were transcriptional inducers of HBD-2 in cultured chondrocytes and cartilage explants in vitro, as measured by real-time RT-PCR and ELISA. These results illustrate the induction of HBD-2 in osteoarthritic cartilage and suggest that it is a further factor in the pathogenesis of OA. However, further studies are required to elucidate the role played by HBD-2 in osteoarthritic cartilage.

Methodik. Zwischen 1980 und 1996 wurden 23 Patienten mit Talusluxationsfrakturen behandelt. Unfallursache war in 61% der Falle ein Verkehrsunfall, in 22% ein Absturz. 5 Patienten (22%) erlitten offene Verletzungen, 2 (9%) entwickelten fruhzeitig ein Kompartmentsyndrom des Fuses und 11 der Verletzten (48%) waren polytraumatisiert. Die Verletzungen wurden nach Hawkins und Marti/Weber klassifiziert. Alle Talusluxationsfrakturen wurden operativ stabilisiert durch Schraubenosteosynthese, Spickdrahttransfixation und/oder Fixateur externe. 15 der Unfallverletzten (65%) wurden im Mittel 5 Jahre nach dem Trauma klinisch und radiologisch untersucht. Ergebnis. Nach dem Kieler Fusscore hatten 4 Patienten ein exzellentes und 3 ein gutes Ergebnis. Bei 5 Patienten mit befriedigendem, 2 mit ausreichendem und 1 mit schlechtem Resultat lagen in 50% zusatzliche ipsilaterale Extremitatenverletzungen vor. In 48% der Falle war die anatomische Rekonstruktion aufgrund knocherner Destruktionen unzureichend. Schlussfolgerungen. Die individuelle Prognose wird durch peritalare Arthrosen (73%) sowie die Talusnekrose (39%) bestimmt. Auch geringer Dislokationsgrad, sofortige Reposition und stabile Osteosynthese konnen diese Komplikationen nicht immer verhindern.

Diskussionen uber die Behandlung von Fersenbeinfrakturen verlaufen weiterhin kontrovers. Um Weichteilprobleme zu minimieren, wurde in der Klinik fur Unfallchirurgie der Universitat Kiel eine optimierte Montageform eines Fixateur externe zur primaren Frakturbehandlung entwickelt. Verwendet wird eine bilaterale Rahmenkonstruktion mit Schanz-Schraubenfixierung in der Tibia und im Tuber calcanei. Sie ermoglicht eine effiziente Reposition uber Ligamentotaxis, eine stabile Fixation und vor allem die aktive Bewegung im OSG. Es wurden 40 Patienten mit 45 Kalkaneusfrakturen mit dem Fixateur externe stabilisiert; 25 Frakturen konnten damit ausbehandelt werden. In 20 Fallen erfolgte sekundar ein Verfahrenswechsel zur internen Osteosynthese; 35 Patienten mit 40 Frakturen konnten mit Hilfe des “Kieler Kalkaneusscore” nachuntersucht werden. Patienten nach Definitivversorgung mit dem Fixateur externe weisen im Mittel ein besseres Spatergebnis auf als Patienten mit spaterem Verfahrenswechsel. Mit der vorgestellten besonderen Montageform ist eine definitive Behandlung des Fersenbeinbruchs moglich. Die Indikation liegt in der ersten Notfallbehandlung von Frakturen mit Weichteilproblemen.

Although children are injured everyday through accidents, road traffic, leisure or sports activities, internal lesions of the knee joint due to the trauma are rare. Diagnose and therapy follow rather empirical than analytical patterns. A retrospective, controlled study evaluates and recommends ways of treatment. Traumatic internal lesions of the knee where analysed in 76 children up to age 16. The pattern of injury changed with increasing age, the trauma remaining the same. Most common where injuries to the anterior cruciate ligament (ACL). Main cause where sports activities. Operative treatment seems to be the appropriate treatment. Osseous avulsions of the cruciates and collateral ligaments showed good results after transosseous refixation with a suture. Suturing of intraligamentous ACL-ruptures as well as patellar ligaments plasty showed unsatisfactory results. Secondary lesions due to instability of the knee where also observed in children. Children cannot self estimate the severity of the injury so subjective statements are insecure. Trauma, surgery, pain and immobilisation cause a marked malfunction of the sensor-motor system which is effectively treated by physiotherapy.

PURPOSE: An analysis of the CT morphology of fractured thoraco-lumbar vertebrae after treatment with internal fixation and transpedicular spongiosaplasty (SP). MATERIAL AND METHOD: 30 patients were examined following trauma and surgery after about 12 and 30 months by means of CT. The following were evaluated: width of the spinal canal; height of the vertebra and intervertebral space; degree of kyphosis; position, size and appearance of the SP and of the vertebral body. RESULTS: The width of the spinal canal was reconstituted in 91%; in 83% the anterior vertebral margin and in 35% the intervertebral space was reduced. A kyphosis of 8.9 degree was found on the followup examination. The SP showed a reduction in size (18/30) or could no longer be defined (6/30). Hypodense areas (28/30) with cavitation (12/30) were found in the vertebral body and the SP could be identified by a sclerotic margin (22/30). CONCLUSION: Treatment by this form of therapy was successful, reaction of the vertebral body against the spongiosaplasty could be identified.

Background Patients with human immunodeficiency virus (HIV) infection and latent tuberculosis are at substantial risk for the development of active tuberculosis. As a public health measure, prophylactic treatment with isoniazid has been suggested for HIV-infected persons who have anergy and are in groups with a high prevalence of tuberculosis. Methods We conducted a multicenter, randomized, double-blind, placebo-controlled trial of six months of prophylactic isoniazid treatment in HIV-infected patients with anergy who have risk factors for tuberculosis infection. The primary end point was culture-confirmed tuberculosis. Results The study was conducted from November 1991 through June 1996. Over 90 percent of the patients had two or more risk factors for tuberculosis infection, and nearly 75 percent of patients were from greater New York City. After a mean follow-up of 33 months, tuberculosis was diagnosed in only 6 of 257 patients in the placebo group and 3 of 260 patients in the isoniazid group (risk rati...

Sprungbeinverletzungen sind seltene, aber schwere Verletzungen. Aufgrund der Anatomie handelt es sich meist um Gelenkfrakturen. Talushalsfrakturen machen etwa 50% aller Sprungbeinbruche aus, es folgen die Korpusfraktur und die seltene Taluskopffraktur. Die sich aus Anamnese, Unfallmechanismus und klinischem Erscheinungsbild ergebende Verdachtsdiagnose Taluslasion wird durch Rontgenaufnahmen gesichert. Die CT dient zur Klarung des operativen Zugangs, der Frakturpathologie, der mehrdimensionalen Nachverarbeitung sowie der postoperativen Kontrolle. Die MRT wird bei persistierenden Beschwerden notwendig. Die Gelenkfrakturen sollten moglichst fruhzeitig behandelt werden, um die haufige Komplikation Nekrose gering zu halten. Es konnen jedoch keine Aussagen zur Wahrscheinlichkeit des Auftretens derselben gemacht werden. Die vielfaltigen Therapiemoglichkeiten werden dargestellt. Hinsichtlich der Nachbehandlung streben wir eine moglichst fruhe funktionelle Therapie an.

In this study we estimated occurrence of the booster effect in a population infected with the human immunodeficiency virus (HIV) and assessed the relation between the booster effect, T-lymphocyte CD4 cell counts, tuberculosis risk categories, and HIV exposure categories. Patients were recruited from 13 participating sites of the Terry Beirn Community Programs for Clinical Research on AIDS (CPCRA). A two-stage tuberculin skin test was applied to 709 HIV-infected patients using the Mantoux method. An induration reading5 mm on the first test andor = 5 on the second skin test defined the booster effect. Overall, 18 patients, or 2.7% (95% confidence interval, 1.6 to 4.2) experienced the booster effect. Boosted responses were seen in eight (2.1%) anergic patients, six (4.5%) nonanergic patients, and four (2.5%) with anergy status unknown. Boosting was noted in 1 of the 131 women enrolled. Age, race, CD4 cell count, injection drug use, anergy status, tuberculosis risk categories, and HIV exposure categories were not predictive of boosting. The booster effect occurs in a small percentage of HIV-infected patients tested, thus identifying small numbers of patients with latent tuberculosis infection. The two-stage procedure is probably of limited value in the diagnosis of latent tuberculosis in HIV-infected persons.

Members of the fibroblast growth factor (FGF) family have been associated with tumor progression and angiogenesis, though the mechanism through which they affect the progression of breast cancer remains elusive. We recently showed that progestins increase the production of the potent angiogenic factor VEGF in an in vivo BT-474 human breast cancer cell-derived xenograft model. In this study we sought to determine the effect of progesterone (P) on regulation of specific FGF family members (FGF-2, FGF-4 and FGF-8) in the same model. Using immunohistochemistry we found that treatment with P significantly reduced FGF-2 and FGF-8 levels, while modestly increasing the levels of FGF-4 in tumors collected at the termination of the study or soon after P treatment began. The in vivo observations with FGF-2 were confirmed in cultured BT-474 cells, though the P-mediated reduction in FGF-2 was not blocked by the anti-progestin RU-486, suggesting that classical progesterone receptors (PR) are not involved in FGF-2 down-regulation. Also, P did not affect levels of FGF-2 mRNA in BT-474 cells, indicating that P exerts its effects on FGF-2 post-transcriptionally. Our observations suggest that the in vivo stimulation of BT-474 cell growth by P is associated with down-regulation of FGF-2 and FGF-8. Furthermore, since FGF-4 levels increased during P-treatment, FGF-4 may be required for tumor growth and maintenance and might therefore be a potential therapeutic target through which to suppress P-dependent tumor growth.

To determine the effects of varying inspired O2 on positive radial acceleration (+Gz; i.e., head-to-foot inertial load) duration tolerance, seven men were exposed to the +4.5- to +7.0-Gz simulated aerial combat maneuver (SACM) by use of the Armstrong Laboratory (Brooks Air Force Base) centrifuge. Exposures were repeated on different days while subjects breathed gas mixtures of fractional concentration of O2 in inspired air (FIO2) between 0.12 and 0.6. SACM duration tolerance was positively related to inspired O2 of FIO2 between 0.12 and 0.2 but was unchanged at FIO2 between 0.2 and 0.6. SACM exposure decreased arterial O2 saturation and increased heart rates; SACM-induced changes were additive to FIO2 effects. The positive relationship between blood lactate and SACM duration tolerance at all FIO2 indicated an anaerobic component. It is concluded that SACM duration tolerance is limited by reduced FIO2 but not enhanced by hyperoxia. Thus the aerobic component of +4.5- to +7.0-Gz SACM duration tolerance is much greater than previously believed.

In einer retrospektiven Analyse werden 114 Mehrfachverletzte untersucht. Nach Korperregionen getrennt, werden Verletzungsmuster gebildet. Am haufigsten mit insgesamt 61 Fallen finden sich die Verletzungskombinationen Kopf-Skelett-Thorax und Kopf-Skelett-Thorax-Abdomen. Die hochste Letalitat mit 44,8% besteht fur die Verletzung aller 4 Korperabschnitte. Alle anderen Verletzungsmuster haben fast identische Sterberaten. Durch eine additive Bewertung der Einzelverletzungen werden Scorewerte errechnet und den Verletzungsmustern zugeordnet. Nur das Verletzungsmuster Kopf-Skelett-Thorax-Abdomen erreicht durchschnittlich hohere Werte. Eine sichere Risikoabschatzung des Einzelfalles ist lediglich fur sehr niedrige oder sehr hohe Wertebereiche moglich. Auch allein aus der schwersten Einzelverletzung kann eine Risikoabschatzung erfolgen. Vor allem an Kopf, Thorax und Becken wird dies durch die Punktebewertung erfast.

1. 1. The relationship between seasonal changes in environmental temperature and hematological parameters was investigated in mature, single comb white leghorn (SCWL) male chickens. 2. 2. Samples of blood plasma, obtained monthly from two groups of birds over two separate 12 month periods, were analysed for corticosterone (CT), 3,5,3′-triiodothyronine (T3), thyroxine (T4), plasma protein (PP), and packed cell volume (PCV). 3. 3. Statistical analyses revealed that blood plasma concentrations of T3 were significantly correlated negatively with monthly dry-bulb temperatures. 4. 4. There were no consistent or significant relationships between monthly dry-bulb temperature and CT, T4, PP or PCV over the two 12 month periods. 5. 5. The results of this study indicate that blood plasma concentrations of T3 are influenced by season of year in mature, male domestic fowl.

Bei der nach Korperregionen geordneten Erfassung der Verletzungsmuster von 468 Mehrfachverletzten finden sich die Kombinationen Skelett/Thorax und Skelett/Thorax/Abdomen am haufigsten. Eine hohere Letalitat ergibt sich fur die Verletzungskombination Skelett/Abdomen. Bei der Feststellung von Einzeldiagnosen im Kollektiv der 106 Verstorbenen zeigt sich im Vergleich mit der Gruppe der 362 Uberlebenden bei Kopfverletzungen eine signifikante Haufung intrakranieller Blutungen, bei Abdominalverletzungen von Leberrupturen und Magen-Darm-Verletzungen, bei Skelettverletzungen uberraschenderweise von Schulter-, Unterarm-und Unterschenkelbruchen. Wahrend bei Kopf- oder. Abdominaltraumen die Einzeldiagnose die letale Risikogefahrdung wesentlich mitbestimmt, gilt dies fur die Verletzung des Thorax und des Stutz- und Bewegungsapparates nicht. Das Verletzungsmuster kann nicht als Leitlinie bei der Bewertung der Mehrfachverletzung verwendet werden.

The risk of toxoplasmic encephalitis complicating AIDS appears largely limited to those HIV-infected patients with serologic evidence of past Toxoplasma gondii infection and low CD4 lymphocyte counts. The Community Programs for Clinical Research on AIDS has initiated a randomized, placebo-controlled trial to determine if clindamycin or pyrimethamine prophylactic regimens are effective and safe in preventing toxoplasmic encephalitis.

Osteomyelitis often causes functional impairment due to tissue destruction. This report demonstrates a novel previously unappreciated role of osteoblasts. Samples of osteomyelitic bone and bacterially challenged osteoblasts produce increased amounts of antimicrobial peptides in order to combat bacterial bone infection. An osteomyelitis mouse model confirmed the osseous induction of the murine homologue of human beta-defensin-2, suggesting a central role in the prevention of bacterial bone infection. Antimicrobial peptides are effectors of the innate defence system and play a key role in host protection at cellular surfaces. Some of them are produced constitutively, whereas others are induced during infection. Human beta-defensins represent a major subclass of antimicrobial peptides and act as a first line of defence through their broad spectrum of potent antimicrobial activity. The aim of the present in-vitro and in-vivo investigations was to study the expression and regulation of human beta-defensin-2 in the case of bacterial bone infection and to analyse the effects of immunosuppressive drugs on bone-derived antimicrobial peptide expression. Samples of healthy human bone, osteomyelitic bone and cultured osteoblasts (hFOB cells) were assessed for the expression of human beta-defensin-2. Regulation of human beta-defensin-2 was studied in hFOB cells after exposure to bacterial supernatants, proinflammatory cytokines and immunosuppressive drugs (glucocorticoids and methotrexate) and was assayed by enzyme-linked immunosorbent assay. An osteomyelitis mouse model was performed to demonstrate the regulation of the murine homologue of human beta-defensin-2, named murine beta-defensin-3, by real-time reverse transcription-polymerase chain reaction and immunohistochemistry. Healthy human bone and cultured osteoblasts are able to produce human beta-defensin-2 under standard conditions. Samples of infected bone produce higher levels of endogenous antibiotics, such as human beta-defensin-2, when compared with samples of healthy bone. A clear induction of human beta-defensin-2 was observed after exposure of cultured osteoblasts to gram-positive bacteria or proinflammatory cytokines. Additional treatment with glucocorticoids or methotrexate prevented bacteria-mediated antimicrobial peptide induction in cultured osteoblasts. The osteomyelitis mouse model demonstrated transcriptional upregulation of the murine homologue of human beta-defensin-2, namely murine beta-defensin-3, in bone after intraosseous contamination of the tibia. Human and murine bone have the ability to produce broad-spectrum endogenous antibiotics when challenged by micro-organisms in vitro and in vivo. Immunosuppressive drugs, such as glucocorticoids or methotrexate, may increase the susceptibility to bone infection by decreasing antimicrobial peptide expression levels in case of microbial challenge. The induction of human beta-defensin-2 following bacterial contact suggests a central role of antimicrobial peptides in the prevention of bacterial bone infection.

Publisher Summary Gram-positive bacterial infections in mice are among the most common causes of sporadic infections in research colonies, but the lack of recent reports of disease under represents disease prevalence in contemporary research facilities. Clinical expression of infection is typical of pyogenic disease, with clinical signs that range from localized conjunctivitis and dermatitis to fulminate septicemia. Treatment of infections is often instituted to salvage valuable mutant mice until studies are concluded or mice can be rederived. Many gram positive-bacteria that cause disease in mice are commensals on the skin and mucous membranes of other laboratory animals and people. Housing and handling procedures must be implemented to minimize transmission by contact with colonized mice or contaminated fomites, including materials used in experimentation. This chapter discusses gram-positive micrococci and corynebacteria as pathogens of concern to researchers who use mice. Clinical expression of disease is typically pyogenic with micrococcal infections and varies from cutaneous hyperkeratosis to septicemia with corynebacterial infections. Treatment of infections may reduce overt disease in affected mice, but elimination of colonization requires rederivation. To raise and maintain mice free from pathogenic gram-positive bacteria, research and resource staff must employ husbandry and handling procedures that minimize contact with contaminated fomites.

Purpose: The combination of abacavir + lamivudine (ABC+3TC) versus didanosine + stavudine (ddI+d4T), each combined with other classes of antiretrovirals (ARVs) in ARV-naive patients, was compared for the combined endpoint of time to plasma HIV RNA >50 copies/mL (at or after the 8-month visit) or death (primary endpoint) in a nested substudy of an ongoing multicenter randomized trial. Method: The substudy enrolled 182 patients; mean HIV RNA and CD4+ cell counts at baseline were 5.1 log10 copies/mL and 212 cells/mm3, respectively. Results: After a median follow-up of 28 months, rates of primary endpoint were 57.2 and 67.8 per 100 person-years for the ABC+3TC and ddI+d4T groups (hazard ratio [HR] = 0.81, 95% confidence interval [CI] 0.58-1.14, p = .23). Conclusion: There was a trend for treatments containing ABC+3TC to be better than treatments containing ddI+d4T with respect to HIV RNA decreases, CD4+ cell count increases, and tolerability.

Vitamin A toxicosis and vitamin E deficiency was diagnosed in a commercial rabbit-breeding colony and was associated with reproductive abnormalities, abortions, and poor survivability of kits in the breeding colony. Paresis and muscular dystrophy were noted in juvenile rabbits. Another group of New Zealand White rabbits from the same commercial colony was used to assess the effect of vitamin E-based therapy on clinical signs, reproduction, and vitamin A and E serum and liver levels. Blood samples were taken before and after dietary changes and vitamin E therapy. Serum vitamin E remained low after feeding a diet containing the recommended levels of vitamin E. Administration of vitamin E for 2 weeks lowered the serum vitamin A levels and increased the vitamin E serum and liver levels. In conclusion, vitamin E therapy appears to be an effective treatment for hypervitaminosis A.

A novel Respirovirus was isolated from nasopharyngeal swab specimens from clinically normal laboratory guinea pigs, and was characterized and named caviid parainfluenza virus 3 (CavPIV-3). The CavPIV-3 is enveloped, is 100 to 300 nm in diameter, and has a characteristic 15-nm-diameter chevron-shaped virus ribonucleocapsid protein. Sequence analysis of the fusion glycoprotein of CavPIV-3 revealed it to be 94% identical to human and guinea pig parainfluenza 3 (PIV-3) viruses and 80% identical to bovine PIV-3. To determine whether CavPIV-3 causes clinical disease in laboratory guinea pigs and to compare the serologic response of guinea pigs to CavPIV-3 and to other paramyxoviruses, an infection study was performed, in which groups of guinea pigs were inoculated with CavPIV-3, Sendai virus, simian virus 5 (SV-5), murine pneumonia virus (PVM), or bovine PIV-3 virus. During the course of the study, guinea pigs were maintained in an infectious disease suite, housed in Micro-Isolator cages, and were only manipulated under a laminar flow hood. Clinical signs of disease were not observed in any of the paramyxovirus-inoculated guinea pigs during the eight-week course of the study, and histologic signs of disease were not evident at necropsy eight weeks after inoculation. Guinea pigs inoculated with CavPIV-3, Sendai virus, PVM, and bovine PIV-3 developed robust homologous or heterologous serologic responses. In contrast, guinea pigs inoculated with SV-5 developed modest or equivocal serologic responses, as assessed by use of an enzyme-linked immunosorbent assay. Further, use of the SV-5 enzyme-linked immunosorbent assay resulted in the highest degree of non-specific reactivity among all of the paramyxovirus assays. In summary, CavPIV-3 is a novel guinea pig Respirovirus that subclinically infects laboratory guinea pigs, resulting in a robust serologic response, but no observed clinical or histologic disease. The CavPIV-3 fusion glycoprotein gene sequence is available from GenBank as accession No. AF394241, and the CavPIV-3 virus is available from the American Type Culture Collection as accession No. DR-1547.

Mice minute virus (MMV) and mouse parvovirus (MPV) type 1 are the two parvoviruses known to naturally infect laboratory mice and are among the most prevalent infectious agents found in contemporary laboratory mouse colonies. Serologic assays are commonly used to diagnose MMV and MPV infections in laboratory mice; however, highly accurate, high-throughput serologic assays for the detection of MMV- and MPV-infected mice are needed. To this end, the major capsid viral protein (VP2) genes of MMV and MPV were cloned and MMV recombinant VP2 (rVP2) and MPV rVP2 proteins were expressed by using a baculovirus system. MMV rVP2 and MPV rVP2 spontaneously formed virus-like particles that were morphologically similar to empty parvovirus capsids. These proteins were used as antigens in enzyme-linked immunosorbent assays (ELISAs) to detect anti-MMV or anti-MPV antibodies in the sera of infected mice. Sera from mice experimentally infected with MMV (n= 43) or MPV (n= 35) and sera from uninfected mice (n= 30) were used to evaluate the ELISAs. The MMV ELISA was 100% sensitive and 100% specific in detecting MMV-infected mice, and the MPV ELISA was 100% sensitive and 98.6% specific in detecting MPV-infected mice. Both assays outperformed a parvovirus ELISA that uses a recombinant nonstructural protein (NS1) of MMV as antigen. The MMV rVP2 and MPV rVP2 proteins provide a ready source of easily produced antigen, and the ELISAs developed provide highly accurate, high-throughput assays for the serodiagnosis of MMV and MPV infections in laboratory mice.

Two from a group of approximately 50 C.B-17 scid-bg mice were examined because of lethargy, dehydration, and rough coat. Three months prior to development of clinical signs of disease, mice of this study had been surgically implanted with fetal bovine liver, thymus, and lymph node. At necropsy, marked splenomegaly and mild hepatomegaly were observed in both animals. Large areas of necrosis and inflammation, with associated intracytoplasmic granular basophilic inclusions, were observed in histologic sections of multiple organs. Aerobic and anaerobic culturing of the liver yielded negative results. Six months after the initial case, four more reconstituted scid-bg mice from a different fetal donor had identical clinical, gross, and histologic signs of disease. To determine whether the basophilic inclusions represented an infective agent, 4-month-old immune-naive C.B-17 scid-bg mice were inoculated intraperitoneally with a liver and spleen homogenate from an affected mouse. Two weeks after inoculation, mice developed clinical signs of disease and lesions identical to those seen in the signal mice. On ultrastructural examination of the liver, pleomorphic bacteria were found in large cytoplasmic vacuoles of hepatocytes. Bacterial DNA was amplified from the liver, using primers that amplify a segment of the 16S rRNA gene from many bacterial species. Sequencing of the polymerase chain reaction (PCR) product revealed gene sequence identical to that of Coxiella burnetii, the agent of Q-fever. These results highlight the need to consider infective agents of the donor species when working with xenografted animals.

Several rodent helicobacters have been associated with chronic active hepatitis or inflammatory bowel disease. Severe combined immunodeficient (SCID) mice appear to be inherently susceptible to disease attributable to these emerging pathogens. With the advent of polymerase chain reaction (PCR) analysis, it has become clear that several as yet unidentified Helicobacter species may also colonize rodents, but their capacity to cause disease is unknown.A Helicobacter species isolated from feces of a BALB/c mouse and provisionally named "H. typhlonicus" was used to inoculate helicobacter-free 4-week-old SCID mice (n = 11 males and 11 females). At various weeks after inoculation, mice were sacrificed and liver and intestinal specimens were collected for histologic examination and PCR analyses.The C.B-17 scid/scid mice inoculated with "H. typhlonicus" developed moderate to severe proliferative typhlocolitis, similar to that seen in SCID mice infected with H. hepaticus or H. bilis. However, in contrast to mice infected with H. hepaticus or H. bilis, lesions of chronic active hepatitis were not detected in mice inoculated with "H. typhlonicus." A similar disease syndrome developed in SCID mice cohabitated with B6D2F1 mice naturally infected with a novel Helicobacter species that was genetically identical to "H. typhlonicus.""Helicobacter typhlonicus" joins a growing list of helicobacters that are capable of inducing enteric disease in immunodeficient mice.

Helicobacter bilis is a recently identified species that colonizes the intestine and liver of mice. In immunocompetent mice, infections have been associated with mild hepatitis, and in immunocompromised mice, inflammatory bowel disease has been induced by intraperitoneal inoculation of the organism. We report inoculation of 6-week-old C.B-17 scid/scid mice by gastric gavage with approximately 10(7) H. bilis colony-forming units. Groups of mice were euthanized and necropsied 12, 24, and 36 weeks after inoculation. Mild to moderate proliferative typhlitis was evident in all mice at 12 and 36 weeks after inoculation and in most mice 24 weeks after inoculation. Mild to severe chronic active hepatitis was detected in 10 of 10 male mice and 3 of 10 female mice. These results indicate that H. bilis can cause moderate to severe enterohepatic disease in immunocompromised mice.

Mouse monoclonal antibodies (MAbs) developed to a rat isolate (R-3) of cilia-associated respiratory (CAR) bacillus were used to assess antigenic relationships among three rat and five rabbit CAR bacillus isolates. Evaluation of MAbs by enzyme-linked immunosorbent assays (ELISAs) indicated that 87 of 241 hybridomas secreted CAR bacillus-reactive antibodies that could be grouped into four major groups. Group-I MAbs reacted with epitopes expressed by all CAR bacillus isolates and at least two or more nonrelated species of bacteria. Group-II, -III, and -IV MAbs reacted with only one or more of the rat CAR bacillus isolates; no MAbs reacted only with rat and rabbit CAR bacillus isolates. Western blot analyses indicated that 41-, 50-, and 105-kDa peptides of rat CAR bacillus isolates expressed rat CAR bacillus group- and isolate-specific epitopes. Hyperimmune anti-CAR bacillus antiserum and serum specimens from a CAR bacillus histologically positive mouse and rat also reacted with the 41-, 50-, and 105-kDa peptides. Sera from CAR bacillus histologically negative rats did not react with these peptides. These results suggest that the 41-, 50-, and 105-kDa peptides may represent suitable antigens for development of a specific ELISA for detection of rodent CAR bacillus infections. Furthermore, these data indicate that use of crude CAR bacillus preparations for either rat or rabbit CAR bacillus ELISAs is inappropriate.

Hyperplastic goiter was diagnosed during routine health monitoring of a closed Syrian hamster colony (SG). Adult and juvenile hamsters were affected at a prevalence of 45%. Histologic examination of the enlarged thyroid gland revealed marked follicular cell hyperplasia. Because prevalence of thyroid hyperplasia in this colony exceeded the 6 to 7% prevalence expected in aged hamsters, additional studies were performed to investigate the pathogenesis of this condition. Juvenile male SG hamsters and age- and sex-matched Syrian hamsters that did not have increased prevalence of goiter were obtained from an unrelated source (Fredrick Cancer Research and Development Center [FCRDC]). The thyroid glands of hamsters were evaluated by 123I radionuclide imaging. Eight of 18 SG hamsters and none of the FCRDC hamsters had a diagnosis of enlarged thyroid gland. Serum baseline and post-thyrotropin thyroxine concentrations in SG hamsters were not statistically different from those in FCRDC hamsters. To investigate whether diet played a role in development of hyperplastic goiter, for 6 months 15 FCRDC hamsters were fed the diet that had been fed to SG hamsters (mouse breeder diet), and five were fed a control diet. To determine whether dietary change would result in resolution of goiter, affected SG hamsters were fed a control diet for 3 months. At the end of each feeding trial, thyroid gland uptake of 123I was reevaluated. The amount of 123I taken up by the thyroid glands of FCRDC hamsters fed the mouse breeder diet was not significantly different from that of controls. In contrast, thyroid gland uptake of 123I remained high for all affected SG hamsters fed the control diet. On the basis of results of these investigations, diet was ruled out as the cause of goiter. Also, a diagnosis of euthyroid hyperplastic goiter was made for the SG hamsters. A genetic cause is suspected to play a role in the increased prevalence of goiter in SG hamsters.

Candidiasis is a frequent complication of infection with the human immunodeficiency virus (HIV); however, few data exist about the natural history, prevention, and treatment of mucosal candidiasis in women.To evaluate the safety and effectiveness of weekly fluconazole prophylaxis for mucosal candidiasis in women infected with HIV.Randomized, double-blind, placebo-controlled trial.14 sites participating in the Community Programs for Clinical Research on AIDS (CPCRA).323 women with HIV infection and CD4+ cell counts of 300 cells/mm3 or less.200 mg of fluconazole per week or placebo. Open-label fluconazole for candidiasis prophylaxis was permitted after two oropharyngeal or vaginal episodes or one esophageal episode.Development of mucosal candidiasis, clinical and in vitro resistance of Candida species to fluconazole, survival, and adverse events.After a median follow-up of 29 months, 72 of 162 patients receiving fluconazole and 93 of 161 patients receiving placebo had at least one episode of candidiasis (relative risk [RR], 0.56 [95% Cl, 0.41 to 0.77); P0.001). Weekly fluconazole was effective in preventing oropharyngeal candidiasis (RR, 0.50 [Cl, 0.33 to 0.74]; P0.001) and vaginal candidiasis (RR, 0.64 [Cl, 0.40 to 1.00]; P = 0.05) but not esophageal candidiasis (RR, 0.91 [Cl, 0.48 to 1.72]; P0.2). Relative risks were similar for women who had a history of mucosal candidiasis (RR, 0.5 [Cl, 0.35 to 0.75]) and those who did not (RR, 0.69 [Cl, 0.35 to 1.34]). Absolute risk reduction for patients with a history of infection was 25.6 per 100 person-years, which is more than twice the reduction of 11.2 per 100 person-years seen in patients with no history of infection. This difference reflects the higher risk of patients who previously had an infection. Candida albicans was not usually resistant to fluconazole in vaginal specimens in clinical or in vitro settings; such resistance occurred in less than 5% of patients in each group.Weekly fluconazole (200 mg) seems to be safe and effective in preventing oropharyngeal and vaginal candidiasis. This regimen has a useful role in the management of HIV-infected women who are at risk for recurrent mucosal candidiasis.

Infection with human immunodeficiency virus (HIV) has been associated with increased rates of single- and multidrug-resistant (MDR) tuberculosis in the New York City area. In order to examine the relationship of HIV infection to drug-resistant tuberculosis in other selected regions of the United States, we established a registry of cases of culture-proven tuberculosis. Data were collected from sites participating in an NIH-funded, community-based HIV clinical trials group. All cases of tuberculosis, regardless of HIV status, which occurred between January 1992 and June 1994 were recorded. Overall, 1,373 cases of tuberculosis were evaluated, including 425 from the New York City area, and 948 from seven other metropolitan areas. The overall prevalence of resistance to one or more drugs was 20.4%, and 5.6% of isolates were resistant to both isoniazid and rifampin (MDR). In the New York City area, HIV-infected patients were significantly more likely than persons not known to be HIV-infected, to have resistance to at least one drug (37% versus 19%) and MDR (19% versus 6%). In other geographic areas, overall drug resistance was 16%, and only 2.2% of isolates were MDR. In multiple logistic regression analyses, HIV infection was shown to be a risk factor for drug-resistant tuberculosis, independent of geographic location, history of prior therapy, age, and race. We concluded that HIV infection is associated with increased rates of resistance to antituberculosis drugs in both the New York City area and other geographic areas. MDR tuberculosis is occurring predominantly in the New York City area and is highly correlated with HIV infection.

Clostridium piliforme infection (Tyzzer's disease) was diagnosed in a colony of nude mice. Because spontaneous Tyzzer's disease had not been reported in nude mice, a study was undertaken to better define the clinicopathologic features of this disease outbreak. Sixty homozygous nude (nu/nu) females, 10 nu/nu males, and 10 heterozygous nude (nu/+) females were observed for signs of disease. Over a 3-month period, 43% of the nu/nu mice died or manifested clinical signs of disease and were euthanized, but nu/+ mice remained healthy. Clinical signs of disease were infrequently observed in nu/nu mice and, when evident, were followed by rapid deterioration and death. Gross and histologic lesions, including severe hepatic and intestinal necrosis associated with C. piliforme, were observed only in clinically affected animals. Clostridium piliforme isolated from diseased livers had marked cytotoxicity in in vitro assays. This outbreak is unique in that, contrary to a previous experimental report, nu/nu mice had increased susceptibility to Tyzzer's disease, suggesting that T cells may play an important role in host defenses against C. piliforme infection. In addition, this is the first report of a toxigenic isolate of C. piliforme recovered from mice. The cytotoxin produced by the isolate may have contributed to the severity of clinical disease and lesions.

Autoimmune MRL/ lpr mice were treated for 12–14 weeks with anti-CD4 monoclonal antibody to define the role of CD4 + T cells in the pathogenesis of the inflammatory central nervous system (CNS) lesions, arthritis and sialadenitis characteristics of the strain. Anti-CD4 therapy effectively prevented the development of CNS lesions and arthropathic changes. Marked depletion of CD4 + T cells was documented in the mononuclear cells infiltrating the major salivary glands but the severity of sialadenitis was significantly increased by chronic anti-CD4 immunotherapy. This dissociation between beneficial and harmful effects of anti-CD4 treatment in the MRL/ lpr mouse suggests that the net regulatory effect of CD4 + T cells on the underlying autoimmune-mediated inflammatory process may be positive or negative depending on the organ system involved. The pathogenetic mechanisms of inflammation and tissue destruction in this model of systemic autoimmune disease are in some instances target organ-specific.

The cilia-associated respiratory (CAR) bacillus is an unclassified, gram-negative bacterium that has been implicated as an etiologic agent of respiratory tract disease in laboratory rodents. A morphologically and antigenically similar organism has been identified in rabbits and is thought to be a related bacterium, although clinical signs of disease and histologic lesions are absent in infected rabbits. To compare the pathogenicity of rat- and rabbit-origin CAR bacillus isolates in rabbits, neonatal rabbits were experimentally infected with CAR bacillus isolates obtained from an infected rat and rabbit. Rabbits experimentally inoculated with rabbit-origin CAR bacillus had a nasal discharge, seroconverted and developed histologic lesions, whereas rabbits inoculated with rat-origin CAR bacillus seroconverted but did not have evidence of colonization of the respiratory tract. The CAR bacillus isolates were further examined at the genetic level by sequencing 1,261 base pairs of the 16S rRNA gene from six CAR bacillus isolates obtained from infected rabbits. A consensus sequence was obtained and compared with the analogous gene sequence data from rat-origin CAR bacillus isolates. Results indicated that these two organisms are distinctly different, with only 48.8% sequence homology. Comparison of the rabbit-origin 16S rRNA gene sequence with the database Genbank indicated that the organism is most closely related to members of the genus Helicobacter. Bacteria with the highest percentage of similarity with the rabbit-origin CAR bacillus were Helicobacter sp. strain Seymour and H. felis, with 91.1 and 90.8%, respectively. Findings of this study indicate that CAR bacillus isolates from rats and rabbits are host-specific and are different bacteria that belong to distinct genera.