A microPET study of the regional distribution of [11C]-PK11195 binding following temporary focal cerebral ischemia in the rat. Correlation with post mortem mapping of microglia activation.

BACKGROUND: Post-stroke microglial activation (MA) may have both neurotoxic and pro-repair effects, particularly in the salvaged penumbra. Mapping MA in vivo is therefore an important goal. 11C-PK11195, a ligand for the 18 kDa translocator protein, is the reference radioligand for MA imaging, but a correlation between the regional distributions of in vivo tracer binding and post mortem MA after stroke, as assessed with PET and immunohistochemistry, respectively, has not been demonstrated so far. Here we performed 11C-PK11195 microPET in a rat model previously shown to induce extensive cortical MA, and determined the correlation between 11C-PK11195 and immunostaining with the CD11 antibody OX42, so as to verify the presence of activated microglia, in a template of PET-resolution size regions-of-interest (ROIs) spanning the whole affected hemisphere. METHODS: Adult spontaneously hypertensive rats underwent 45 min distal middle cerebral artery occlusion and 11C-PK11195 PET at Days 2 and 14 after stroke according to a longitudinal design. Following perfusion-fixation at Day 14, brains were removed and coronally cut for OX42 staining. 11C-PK11195 binding potential (BPND) parametric maps were generated, and in each rat both BP(ND) and OX42 (intensity×extent score) were obtained in the same set of 44 ROIs extracted from a cytoarchitectonic atlas to cover the whole hemisphere. Correlations were computed across the 44 ROIs both within and across subjects. RESULTS: Significant BPND increases were observed in both the infarct and surrounding areas in all rats at day 14; less strong but still significant increases were present at day 2. There were highly significant (all p<0.001) positive correlations, both within- and across-subjects, between day 14 BPND values and OX42 scores. CONCLUSIONS: The correlation between Day 14 11C-PK11195 and OX42 across the affected hemisphere from the same brain regions and animals further supports the validity of 11C-PK11195 as an in vivo imaging marker of MA following stroke. The finding of statistically significant increases in 11C-PK11195 as early as 48 h after stroke is novel. These results have implications for mapping MA after stroke, with potential therapeutic applications.

Mapping selective neuronal loss and microglial activation in the salvaged neocortical penumbra in the rat.

Rescuing the ischemic penumbra from infarction is the mainstay of acute stroke therapy. However, the rescued penumbra may be affected by selective neuronal loss (SNL) and microglial activation (MA), which may hinder functional recovery and hence represent potential new therapeutic targets. Imaging them in vivo is currently attracting considerable interest, but relevant rat models are needed to underpin methods development and validation. Although striatal SNL/MA is well described following proximal MCA occlusion (MCAo), neocortical SNL/MA is still poorly characterized, yet has greater clinical relevance. This study aimed to assess the distribution and intensity of neocortical SNL and MA in a distal clip MCAo model known to cause severe neocortical ischemia. Spontaneously hypertensive rats were subjected to 45 min distal MCAo with ipsilateral common carotid artery occlusion. At day 14, post mortem SNL and MA were mapped using NeuN and OX42 immunohistochemistry, respectively. In a separate group, cerebral blood flow (CBF) was mapped during MCAo using (14)C-iodoantipyrine autoradiography. Values for SNL, MA, and CBF were obtained in the same set of anatomical ROIs covering the cortical MCA territory. Extensive SNL and MA affected the non-infarcted MCA cortex, adopting a well-defined regional distribution and a striking patchy/pseudo-columnar pattern. Regional intensities of SNL and MA were strongly inter-correlated, and also strikingly related to occlusion CBF, showing sharp rises for CBF <40%, i.e. the penumbra threshold. This rat model may be useful in providing in vitro reference for studies aiming to validate novel imaging tracers of SNL and MA in vivo.

Reduced structural proteins in the conjunctival epithelium in allergic eye disease.

AIMS: Allergic eye disease affects up to 20% of the population with varying severity. The conjunctival epithelium plays a key role in allergic eye disease. The purpose of this study was to determine whether the conjunctival epithelium is abnormal in allergic eye disease. METHODS: Conjunctival biopsy samples were taken from patients with seasonal allergic conjunctivitis (SAC) 'in' and 'out of season' and nonatopic control subjects. Specimens were fixed in glycol methacrylate, 2 microm serial sections cut and Image-J used to assess the sites and areas of immuno-staining. RESULTS: E-cadherin, CD44, keratins K5/6, K8, K13, K14, K18 and pan-keratin immuno-staining were all significantly lower in patients 'out of season' compared with normal controls. No structural differences in the epithelium were observed between the two groups. The epithelium of patients 'in season' was thicker and immuno-staining of the above markers similar to controls. CONCLUSIONS: The expression of a wide spectrum of epithelial cell adhesion proteins and cytoskeletal elements is downregulated in the conjunctiva of SAC patients 'out of season' compared with normal controls. We suggest that this could have an important impact on the ability of the epithelium to protect itself against allergen penetration, potentially influencing the development and course of allergic eye disease and offering a novel area for therapeutic control.

More and earlier surfactant for preterm infants.

Surfactant administration to infants born at less than 32 weeks gestation was compared between two time periods (1 April 1994 to 31 March 1996 and 1 April 1999 to 31 March 2001). Overall administration increased significantly from 41% to 54%, and within one hour of birth from 13% to 60%. Regional data collection and feedback helps promote quality improvement and implementation of published evidence and guidelines.

Self-administration of epinephrine in children: a survey of current prescription practice and recommendations for improvement.

The prevalence of peanut allergy is increasing rapidly and many children are now prescribed self-injectable epinephrine as part of their management. We aimed to examine the current extent of self-injectable epinephrine dispensing to children in the Eastern Health and Social Services Board (EHSSB), Northern Ireland, including indications for prescription, investigations performed, information and training provided and actual usage. Dispensing records held by the EHSSB were examined for the period May to August 1998. All general practitioners prescribing 'Epipens' during this period were contacted and asked to identify the patient and provide contact details. Information was gathered using postal questionnaires sent to General Practitioners and parents. A total of 104 'Epipen' prescriptions were dispensed. Thirty-seven (36%) general practitioners responded to the initial questionnaire; of these 36 (35%) were suitable for analysis. Thirty-four parents were then contacted; 28 (82%) returned questionnaires were reviewed. The commonest indication for 'Epipen' prescription was peanut allergy (32 of 36 (89%) general practitioner responses; 25 of 28 (89%) parent responses). Twenty-six (72%) children had been seen by a specialist; all except one had either blood or skin tests. Six of the remaining eight children had no investigations. General practitioners reported 14 (39%) parents to have basic life support training, compared with six (21%) parents. Eighteen (64%) parents had been given written information regarding their child's allergy, nine (32%) had been referred to a dietician and seven (25%) children wore a medical warning bracelet. The Epipen had been used by three children; all three had multiple food allergies. This study has identified a great variability in the management of children with allergy including the need for specialist referral, further investigation, written allergy advice, referral to a dietician and formalised training in basic life support and administration of epinephrine. It suggests a lack of consensus amongst health care professionals as to the best practice in the management of potentially life threatening food allergy and indicates, at least, a need for better multidisciplinary communication.

Prevention of venous thromboembolism in Wales: results of a survey among general surgeons.

OBJECTIVE: To examine the current attitudes towards the prevention of venous thromboembolism among a cohort of surgeons. DESIGN: A postal survey, comprising a questionnaire covering various aspects of venous thromboembolism prophylaxis was sent to all (n=84) consultant general surgeons in Wales. RESULTS: Replies were received from 57 surgeons (68%), all of whom routinely used prophylaxis, the most frequent modalities used being heparin (100%) and graded compression stockings (79%). A combination of physical and pharmacological methods was used by over 89% of surgeons, with 60% starting prophylaxis more than two hours before operation. All surgeons continued prophylaxis after surgery, 53% until patients were mobile, 45% until they were discharged, and one surgeon continued prophylaxis for seven days after discharge. The thrombosis risk factors considered most important by surgeons when deciding about prophylaxis were (i) a previous history of venous thromboembolism, (ii) hypercoagulability, and (iii) malignancy. CONCLUSIONS: This study confirms that Welsh surgeons conform to standard methods, but also highlights some uncertainties that are present in current surgical practice. Those who responded all routinely used prophylaxis, the timing of which was variable. The main risk factors identified when considering prophylaxis were previous history of deep vein thrombosis/pulmonary embolism, hypercoagulability, and the presence of malignancy. Suggestions for future practice are made.

Fixation of distal femoral fractures above total knee arthroplasty utilizing the Less Invasive Stabilization System (L.I.S.S.).

Supracondylar femoral fractures above total knee arthroplasty remain a treatment challenge. Complication rates as high as 30% are associated with both nonoperative and operative treatment. Conventional plate fixation and rigid intramedullary nail fixation has improved the treatment of these fractures. However, problems still exist in the setting of a short distal femoral block and/or significant osteoporosis. Less Invasive Stabilization System (L.I.S.S.) fixation has been utilized for the treatment of supracondylar femoral fractures above total knee arthroplasty. Multiple fixed angle screws give optimal fixation around the femoral component. Advantages appear to include maintenance of distal femoral fixation, low infection, and low need for bone grafting.

Activity and expression of the Na(+)/H(+) exchanger in the microvillous plasma membrane of the syncytiotrophoblast in relation to gestation and small for gestational age birth.

The effect of gestational age, low birth weight, and umbilical plasma pH on the activity and expression of the Na(+)/H(+) exchanger in the microvillous plasma membrane (MVM) of the placental syncytiotrophoblast was investigated. MVM were isolated from placentas of fetuses delivered in the first and second trimesters and from appropriately grown for gestational age (AGA) and small for gestational age (SGA) babies born at term. Na(+)/H(+) exchange activity (amiloride-sensitive Na(+) uptake) was higher (p<0.05) in second trimester and term AGA MVM versus first trimester MVM (median [range]: 1.80 [1.01-3.03], 1.72 [1.16-3.15] versus 1.48 [0.92-1.66] nmol/mg protein/30s, respectively, n = 6, 12, and 9). As regards exchanger isoforms, Western blotting showed that NHE1 expression did not change across gestation, but NHE2 and NHE3 expression were lower (p<0.01) in the first and second trimesters than in term AGA MVM. There were no differences in Na(+)/H(+) exchanger activity or in NHE1-3 expression in term AGA MVM versus SGA (n = 11) MVM. There was no correlation between exchanger activity and umbilical artery or vein plasma pH, although with a relatively small number of samples (n = 12 and 15, respectively). We conclude that there is differential regulation of the activity and expression of Na(+)/H(+) exchanger isoforms in the MVM over the course of gestation in normal pregnancy; this is not affected in pregnancies resulting in SGA babies at term.

A cytotoxin-producing strain of Vibrio cholerae non-O1, non-O139 as a cause of cholera and bacteremia after consumption of raw clams.

We report a case of a cholera-like gastroenteritis subsequent with bacteremia in a healthy man following consumption of raw clams. Although we failed to recover the organism from the patient's stool culture, his blood culture was positive for a non-cholera toxin-producing yet cytotoxin-producing non-O1 and non-O139 Vibrio cholerae.

The effects of growth hormone and insulin-like growth factor on the proliferation rate and morphology of RAW 264.7 macrophages.

UNLABELLED: It is well documented that growth hormone (GH) functions to regulate both cell growth and cell number and is considered the master hormone because it affects almost every cell of the body. Growth hormone stimulates the liver to secrete insulin-like growth factor (IGF-1), which is also capable of binding insulin as well as insulin-like binding receptors on the cell surface. It is possible that GH cellular effects are mediated by IGF-1 rather than GH itself. In this study, RAW 264.7 cells were challenged with a high dose of GH (48 ng/microliter), a low of dose GH (4.8 ng/microliter), a high dose of IGF-1 (26 ng/microliter) or low doses of IGF-1 (6.3 ng/microliter) for 24, 48, 72, and 96 hours. Cell number, cell protein concentration, cell damage, and cellular morphology were measured at each time point and compared to untreated RAW 264.7 cells. The results show significant increases in cell number for cells treated with low doses of GH and IGF-1 at 24 hour phase. Cell proliferative effects were also observed at 48 hours in IGF-1 treated cells. Cellular damage (MDA levels) was not statistically significant for any treated group for the entire duration of the experiment. Most notable differences were observed in cellular morphology for both IGF-1 and GH treated cells. IGF-1 resulted in condensation of the nuclear material as early as 24 hours after treatment. IN CONCLUSION: (1) RAW 264.7 responded to both IGF-1 and GH equally (viability and proliferation), and (2) morphological changes were observed in all cells treated with both hormones compared to control group. This study indicates that GH hormone could induce its effect directly or indirectly through IGF-1.

Development of sustained delivery system as a novel technique for tissue culture.

One of the greatest advances for biomedical research has been the use of tissue culture to study the effects and the mode of action of various organic compounds. However, there are myriad problems associated with the classical approach to pharmaceutical studies in tissue culture, such as contamination, the logistical problems inherent in intermittent dosing, and near-lethal drug doses required to achieve effective physiological doses. This has given impetus to a search for better means of effective delivery routes in a tissue culture setting. The specific objectives of this investigation were: (1) to develop a ceramic delivery system for the novel purpose of releasing sustained levels of the anti-cancer drug combination inositol hexaphosphate (IP6) + inositol (I) in a tissue culture setting; (2) to assess the proliferation rate and viability of HTB 122 Intraductile Breast Cancer (IBC) cells exposed to sustained levels of IP6 + I compared to conventional means of drug delivery (dissolved in media); (3) to evaluate the morphological changes associated with this treatment process. Sterile aseptic techniques were employed during the entire study (72 hours of incubation). Data obtained from this investigation suggests the following: (1) TCPL delivery system can be utilized in the tissue culture setting to deliver IP6 + I in a sustained manner for 72 hours; (2) the use of IP6 + I at the physiological dose did not induce any significant change in the biochemical marker (MDA); (3) morphological evaluation of cells treated with sustained delivery resulted in cellular atrophy, as well as fragmentation, compared to the treatment by conventional means and to the sham group (empty capsule); and (4) regardless of route of administration, treatment with IP6 + I resulted in a decrease in total protein content compared to the sham capsules and control groups. In conclusion, this approach is a novel use of ceramic delivery systems in tissue culture that gives breakthrough information for basic research on limiting and eliminating contamination and the logistical problems associated with intermittent dosing in tissue culture.

Abiotrophia species as a cause of endophthalmitis following cataract extraction.

Microorganisms of the genus Abiotrophia, members of the oral flora, are known as important causes of bacterial endocarditis. In this study, we report two individual cases of acute vitreous infection caused by Abiotrophia adiacens and Abiotrophia defectiva approximately a week after cataract extraction. Abiotrophia isolates were recovered by cultivation of vitreous humor on chocolate agar and identified via conventional and API 20 Strep identification systems. An 83-year-old male patient (A) and an 80-year-old female patient (B) demonstrated almost identical symptoms of infectious endophthalmitis manifested as hypopyon and opaque media. The vision of both patients was reduced to detection of hand motion in the left and the right eyes, respectively. An emergency pars plana core vitrectomy was performed, and intraocular antibiotics were administered to each patient, who presented 8 months apart in two different institutions. Patients A and B were treated with an intravitreal injection of vancomycin-amikacin and vancomycin-ceftazidime, respectively, which resulted in complete recovery.

The secreted hepatitis B precore antigen can modulate the immune response to the nucleocapsid: a mechanism for persistence.

The hepatitis B precore Ag (HBeAg) is a secreted nonparticulate version of the viral nucleocapsid hepatitis B core Ag (HBcAg), and its function is unknown. A proportion of HBeAg-specific Th cells evade deletion/anergy in HBeAg-transgenic (Tg) mice and mediate anti-HBe "autoantibody" (autoAb) production after in vivo activation with the appropriate Th cell peptide. This model system was used to determine how secretory HBeAg may effect deletion of Th cells in the periphery. For this purpose, HBeAg-Tg mice were bred with Fas and Fas ligand (FasL)-defective lpr/lpr and gld/gld mutant mice. Fas-FasL interactions mediate activation-induced apoptosis in the periphery. In HBeAg-Tg/+ mice, high-titrated anti-HBe autoAb was produced that was exclusively composed of the IgG1 isotype (i.e., Th2-like profile). In contrast, HBeAg-Tg/lpr and HBeAg-Tg/gld mice produced significantly less anti-HBe autoAb, and the IgG isotype patterns were broadened to include IgG2a, IgG2b and IgG3 as well as IgG1 (i.e., mixed Th1/Th2-like profile). These results suggest that HBeAg-specific Th1 cells are preferentially depleted by Fas-FasL-mediated interactions. The effect of circulating HBeAg on HBcAg-specific Th1 cells was also examined by transferring HBe/HBcAg-specific Th cells into dual HBeAg- and HBcAg-expressing Tg recipient mice. The presence of serum HBeAg ablated the expected Th1-mediated anti-HBc Ab response and shifted it toward a Th2 phenotype. These results suggest that in the context of a hepatitis B viral infection, circulating HBeAg has the potential to preferentially deplete inflammatory HBeAg- and HBcAg-specific Th1 cells that are necessary for viral clearance, thereby promoting hepatitis B virus persistence.

The immune microenvironment of human fracture/soft-tissue hematomas and its relationship to systemic immunity.

The immune environment of human soft-tissue injury is unstudied. We studied fracture soft-tissue hematomas (FxSTH) in 56 patients with high-energy bony fractures. FxSTH serum and mononuclear cells (MNC) as well as fracture patient plasma and blood MNC were studied. Twenty healthy controls donated plasma and MNC. Soluble tumor necrosis factor (TNF)-alpha, interleukin (IL-1 beta, IL-2, 6, 8, 10, 12, and interferon-gamma were studied by enzyme linked immunosorbent assay. Cells were studied by flow cytometry after cell-membrane stains for CD-14, TNF-alpha (mTNF), and human leukocyte antigen-DR, or intracellular stains for TNF (icTNF) and IL-10. Thirty-six patients with Injury Severity Score < 15 were analyzed further to evaluate the effects of isolated fracture on systemic immunity. Cytokines were rarely detectable in control plasma. TNF-alpha, IL-1 beta, IL-2, and interferon-gamma were rarely found in FxSTH serum or fracture patient plasma. All FxSTH sera were rich in IL-6, peaking before 48 hours (12,538 +/- 4,153 vs. 3,494 +/- 909 pg/mL, p = 0.02, U test). In Injury Severity Score < 15, IL-6 was not detectable in most early fracture patient plasma, but rose after 48 hours (p = 0.028). FxSTH serum IL-8 peaked after 48 hours (440 +/- 289 vs. 4,542 +/- 1,219 pg/mL, p = 0.006) and circulating IL-8 appeared after 72 hours. IL-6 and IL-8 showed gradients from FxSTH serum to paired PtS (p < 0.05, Wilcoxon). IL-10 was abundant (884 +/- 229 pg/mL) in FxSTH serum < 24 hours old. FxSTH serum IL-12 peaked late (3,323 +/- 799 pg/mL, day 4-7) then fell (p < 0.001, analysis of variance). Only IL-12 was higher in fracture patient plasma (1,279 +/- 602 pg/mL) than FxSTH serum (591 +/- 327 pg/mL) during the first 48 hours (p = 0.032, U test). On flow cytometry, control monocytes expressed 201 +/- 31 mTNF sites/cell, but icTNF was absent. mTNF was up-regulated after injury more in FxSTH monocytes (3,202 +/- 870 sites/cell) than peripheral blood monocytes (584 +/- 186 sites/cell) (p < 0.05 vs. peripheral blood monocytes by Wilcoxon, p < 0.001 vs. control monocytes by U test). Intracellular IL-10 was abundant in all MNC, but varied widely after injury. Fracture and peripheral blood monocytes expressed far less human leukocyte antigen-DR than control monocytes. Fractures create an inflammatory local environment. Proximal mediators are cell-associated and relatively confined to the wound, but soluble IL-6, IL-8, and IL-10 are abundant and probably exported. Systemic MNC have complex responses to local injuries. These may reflect the combined impact of multiple soluble cytokines initially generated within the wound. FxSTH appear to be a potentially important source of immunomodulatory cytokines in trauma.

The hepatitis B virus core and e antigens elicit different Th cell subsets: antigen structure can affect Th cell phenotype.

Secretion of the hepatitis B virus (HBV) e antigen (HBeAg) has been conserved throughout the evolution of hepadnaviruses. However, the function of this secreted form of the viral nucleoprotein remains enigmatic. It has been suggested that HBeAg functions as an immunomodulator. We therefore examined the possibility that the two structural forms of the viral nucleoprotein, the particulate HBV core (HBcAg) and the nonparticulate HBeAg, may preferentially elicit different T helper (Th) cell subsets. For this purpose, mice were immunized with recombinant HBcAg and HBeAg in the presence and absence of adjuvants, and the immunoglobulin G (IgG) isotype profiles of anti-HBc and anti-HBe antibodies were determined. Second, in vitro cytokine production by HBcAg- and HBeAg-primed Th cells was measured. The immunogenicity of HBcAg, in contrast to that of HBeAg, did not require the use of adjuvants. Furthermore, HBcAg elicited primarily IgG2a and IgG2b anti-HBc antibodies, with a low level of IgG3, and no IgG1 anti-HBc antibodies. In contrast, the anti-HBe antibody response was dominated by the IgG1 isotype; low levels of IgG2a or IgG2b anti-HBe antibodies and no IgG3 anti-HBe antibodies were produced. Cytokine production by HBcAg- and HBeAg-primed Th cells was consistent with the IgG isotype profiles. HBcAg-primed Th cells efficiently produced interleukin-2 (IL-2) and gamma interferon (IFN-gamma) and low levels of IL-4. Conversely, efficient IL-4 production and lesser amounts of IFN-gamma were elicited by HBeAg immunization. The results indicate that HBcAg preferentially, but not exclusively, elicits Th1-like cells and that HBeAg preferentially, but not exclusively, elicits Th0 or Th2-like cells. Because HBcAg and the HBeAg are cross-reactive in terms of Th cell recognition, these findings demonstrate that Th cells with the same specificity can develop into different Th subsets based on the structural form of the immunogen. These results may have relevance to chronic HBV infection. Circulating HBeAg may downregulate antiviral clearance mechanisms by virtue of eliciting anti-inflammatory Th2-like cytokine production. Last, the influence of antigen structure on Th cell phenotype was not absolute and could be modulated by in vivo cytokine treatment. For example, IFN-alpha treatment inhibited HBeAg-specific Th2-mediated antibody production and altered the IgG anti-HBe isotype profile toward the Th1 phenotype.

Fibrous capsule formation in response to ultrahigh molecular weight polyethylene treated with peptides that influence adhesion.

A wide variety of hard tissue substitute materials for reconstruction have been designed for use in dental and orthopaedic procedures. Most of the literature provides information on the osteocompatibility of these materials, and not on soft tissue compatibility. The bone bonding strength of the materials is relatively high, but there seems to be a lack of bonding properties with soft tissue which results in the formation of fibrous encapsulation. This lack of soft tissue bonding can lead to implant failure. Therefore in order to study how artificial materials may remain in the body for long periods of time and remain functional, it is vital to understand the fibrous tissue encapsulation process. Ultrahigh molecular weight polyethylene coated with RGD, RGE, Poly-L-Lysine, or saline was implanted into Sprague Dawley rats either intraperitoneally or subcutaneously for 12 weeks. Aseptic surgical techniques were followed according to standard laboratory procedures. Blood was collected weekly for the first 8 weeks and at 12 weeks, and analyzed for lactate dehydrogenase activity to assess muscle wasting and compatibility. Lactate dehydrogenase activity was elevated at weeks one and two for groups implanted with materials, in comparison to control unoperated animals. From weeks 3-12, the LDH activity was similar in all groups indicating that there was no adverse response to the implanted material. The implants were harvested at the end of 12 weeks, and the fibrous capsule was screened histologically to determine thickness and cell types at the interface. The results clearly showed that the implants that were placed subcutaneously had a less extensive fibrous and vascular tissue formation, than those implanted intraperitoneally. The results also indicated that the location of the implant intraperitoneally, determined the extent of the fibrous tissue formation. The implanted materials coated with RGD and Poly-L-Lysine had thicker fibrous capsule formation than RGE coated implanted at both the subcutaneous and intraperitoneal sites. The results also suggest that the implant-interface contained fewer inflammatory cells when the implants were placed subcutaneously. The overall results indicate that more than surface charge or coating govern the formation of fibrous tissue in vivo.

Biochemical and immunochemical evaluation of tissues and synovial fluid from patients undergoing total joint arthroplasty.

Cytokines are inflammatory mediators responsible for numerous clinical conditions, and are thought to lead to the resorption of bone. Understanding the nature of the cells producing these factors which control the resorption of bone will ultimately lead to a better understanding of why implants fail or integrate. In this study, synovial tissues and synovial fluids were processed for biochemical as well as histochemical and immunohistochemical determination cytokines responsible for bone resorption. The results from this study showed by both quantitative enzyme linked immunoassay (ELISA) and qualitatively by immunohistology a marked increase (twofold) in interleukin-1 (IL-1), and tumor necrosis factor-beta (TNF beta) in synovial tissues in comparison to control tissues of cartilage, ligament and meniscus. Evaluation of tissues both immunochemically and by Hematoxylin and Eosin demonstrated the presence of fibroblast and cells such as macrophages, and multinucleated giant cells in the synovium that are capable of producing bone resorption. Synovial fluid from primary and revision patients were evaluated for TNF beta and IL-1 were not statistically different. Overall, the results indicate that the inflammatory cells of the synovium are secreting factors which may act to mediate aseptic loosening of implants.

Role of B cells in antigen presentation of the hepatitis B core.

The hepatitis B virus (HBV) nucleocapsid or core antigen (HBcAg) is extremely immunogenic during infection and after immunization. For example, during many chronic infections, HBcAg is the only antigen capable of eliciting an immune response, and nanogram amounts of HBcAg elicit antibody production in mice. Recent structural analysis has revealed a number of characteristics that may help explain this potent immunogenicity. Our analysis of how the HBcAg is presented to the immune system revealed that the HBcAg binds to specific membrane Ig (mIg) antigen receptors on a high frequency of resting, murine B cells sufficiently to induce B7.1 and B7.2 costimulatory molecules. This enables HBcAg-specific B cells from unprimed mice to take up, process, and present HBcAg to naive Th cells in vivo and to T cell hybridomas in vitro approximately 10(5) times more efficiently than classical macrophage or dendritic antigen-presenting cells (APC). These results reveal a structure-function relation for the HBcAg, confirm that B cells can function as primary APC, explain the enhanced immunogenicity of HBcAg, and may have relevance for the induction and/or maintenance of chronic HBV infection.

Production of interleukin-10 in human fracture soft-tissue hematomas.

Clinical trauma suppresses immunity and experimental wound fluids have been shown to be immunosuppressive. To ascertain whether human wounds contain immunosuppressive cytokines, we assayed serum from fracture/soft-tissue hematomas (FSTH) of 22 patients for interleukin (IL)-10, transforming growth factor (TGF)-beta 1, and IL-4. Results were correlated to concurrent plasma cytokine concentrations in the same patients and in volunteer plasma. IL-10 was present in high concentration (1376 +/- 539 pg/mL) in all (7/7) FSTH < 24 h old. In FSTH > 24 h old, IL-10 was found intermittently and at lower levels (239 +/- 106 pg/mL, p = .011 vs. FSTH < 24 h old). IL-10 was rarely detectable in fracture patient plasma and never detectable (< 20 pg/mL) in normal plasma. No significant variations of IL-4 or total TGF-beta 1 were found in FSTH or plasma. FSTH are significant potential sources of IL-10 activity in trauma patients, which may be overlooked when only plasma is assayed. The potential for a relationship between cytokines found locally at sites of injury and clinical immune modulation in trauma requires further study.