Validation of a new highly standardised, lab-independent whole-blood leukocyte function assay for clinical trials (ILCS).

Physical stress induced in healthy volunteers by the Loughborough intermittent shuttle test (LIST) was used to validate a newly developed whole-blood cell culture system (Instant leukocyte culture system (ILCS). Exercise induced immune modulation was investigated through measurement of cytokine levels after activating leukocytes in peripheral blood ex vivo using the physiologic stimulant lipopolysaccharide (LPS). LPS induced production of three different cytokines, interferon gamma (IFNgamma), interleukin-6 (IL-6), and interleukin-10 (IL-10). IFNgamma levels were significantly decreased (P = 0.02 and P = 0.001 ) and IL-10 levels significantly increased (P= 0.04 and 0.03) after exercise. LPS induced IL-6 production was only marginally further increased by exercise. In conclusion, the ILCS system provided a reliable ex vivo method, showing common as well as subject specific features in the time course of the immune modulation caused by the LIST protocol. This system will be useful for studies of the elderly, where cytokine standardisation is notoriously difficult.

Plasma cytokine profiles in elderly humans.

It is known that as we age, immune dysregulation often occurs, leading to failing health, and increased susceptibility to a number of different diseases. In this study we have investigated plasma cytokine profiles in order to identify immune markers of ageing. Plasma samples were obtained from 138 participants of the Swedish longitudinal NONA study (aged 86, 90 and 94 years) and 18 healthy Swedish volunteers (aged between 32 and 59 years). Our results show significantly increased levels of the pro-inflammatory cytokine interleukin-6 (P<0.0001) and soluble intercellular adhesion molecule-1 (P<0.0001) in the elderly group. The anti-inflammatory cytokine interleukin-10 did not alter with age whereas active (naturally processed) transforming growth factor-beta levels were significantly (P<0.0001) increased in the elderly group. No difference was observed between males and females. These data suggest that there are measurable changes in cytokine profiles with ageing with increased levels of potentially harmful molecules, which may contribute to immune alterations and declining health in the elderly population.

Influence of age and dietary fish oil on plasma soluble adhesion molecule concentrations.

Soluble forms of intercellular adhesion molecule-1, vascular cell adhesion molecule-1 and E-selectin (termed sICAM-1, sVCAM-1 and sE-selectin respectively) are found in the plasma, and are elevated during inflammatory conditions in which there is increased expression of the cellular forms of the molecules on endothelial and other cells. sICAM-1, sVCAM-1 and sE-selectin concentrations were measured in the plasma of 140 healthy Caucasian subjects aged between 18 and 75 years (100 males/40 females). sICAM-1 concentrations varied between 59.9 and 299.7 ng/ml (median 150 ng/ml), sVCAM-1 concentrations varied between 222.8 and 1672.9 ng/ml (median 662 ng/ml) and sE-selectin concentrations varied between 12.4 and 90.3 ng/ml (median 45.5 ng/ml). There were significant positive linear correlations between age and the plasma concentrations of sICAM-1 (r=0.580; P<0.001) and sVCAM-1 (r=0.392; P<0.001), which were retained when the effects of gender, body mass index and fasting plasma triacylglycerol and total cholesterol concentrations were controlled for. The significant positive linear correlation between age and the plasma concentration of sE-selectin (r=0.234; P=0.027) was lost when other variables were controlled for. Male subjects <40 years of age had significantly lower plasma concentrations of both sICAM-1 and sVCAM-1 than males >55 years of age (both P<0.001), but the difference in plasma sE-selectin concentrations between the age groups did not reach significance (P=0.073). Subgroups of 16 males aged <40 years and 12 elderly subjects (>55 years of age) participated in a doubled-blind, placebo-controlled study of fish oil supplementation over 12 weeks. The level of eicosapentaenoic acid in plasma phospholipids did not change with placebo supplementation, but was significantly increased with fish oil supplementation in both young male and elderly subjects (median increase 200%). sICAM-1, sVCAM-1 and sE-selectin concentrations were unaffected by supplementation with placebo in either young male or elderly subjects. sICAM-1 concentrations were unaffected by fish oil supplementation. sE-selectin concentrations were significantly increased by fish oil supplementation in young males (P=0.043; median increase 38%), but fish oil tended to decrease plasma sE-selectin concentrations in the elderly subjects (P=0.075), with a median decrease of 11%. sVCAM-1 concentrations were unaffected by fish oil supplementation in young males. Fish oil supplementation significantly decreased plasma sVCAM-1 concentrations in the elderly subjects (P=0.043), with a median decrease of 20% (range 16-60%). These observations suggest that fish oil decreases endothelial activation in elderly subjects.

Influence of dietary supplementation with long-chain n-3 or n-6 polyunsaturated fatty acids on blood inflammatory cell populations and functions and on plasma soluble adhesion molecules in healthy adults.

Greatly increasing the amounts of flaxseed oil [rich in alpha-linolenic acid (ALNA)] or fish oil (FO); [rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] in the diet can decrease inflammatory cell functions and so might impair host defense. The objective of this study was to determine the effect of dietary supplementation with moderate levels of ALNA, gamma-linolenic acid (GLA), arachidonic acid (ARA), DHA, or FO on inflammatory cell numbers and functions and on circulating levels of soluble adhesion molecules. Healthy subjects aged 55 to 75 yr consumed nine capsules per day for 12 wk. The capsules contained placebo oil (an 80:20 mix of palm and sunflowerseed oils) or blends of placebo oil with oils rich in ALNA, GLA, ARA, or DHA or FO. Subjects in these groups consumed 2 g ALNA; approximately 700 mg GLA, ARA, or DHA; or 1 g EPA plus DHA (720 mg EPA + 280 mg DHA) daily from the capsules. Total fat intake from the capsules was 4 g per day. None of the treatments affected inflammatory cell numbers in the bloodstream; neutrophil and monocyte phagocytosis or respiratory burst in response to E. coli; production of tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 in response to bacterial lipopolysaccharide; or plasma concentrations of soluble intercellular adhesion molecule-1. In contrast, the ALNA and FO treatments decreased the plasma concentrations of soluble vascular cell adhesion molecule-1 (16 and 28% decrease, respectively) and soluble E-selectin (23 and 17% decrease, respectively). It is concluded that, in contrast to previous reports using higher amounts of these fatty acids, a moderate increase in consumption of long-chain n-6 or n-3 polyunsaturated fatty acids does not significantly affect inflammatory cell numbers or neutrophil and monocyte responses in humans and so would not be expected to cause immune impairment. Furthermore, we conclude that moderate levels of ALNA and FO, which could be incorporated into the diet, can decrease some markers of endothelial activation and that this mechanism of action may contribute to the reported health benefits of n-3 fatty acids.

Manipulation of the type of fat consumed by growing pigs affects plasma and mononuclear cell fatty acid compositions and lymphocyte and phagocyte functions.

To investigate the immunological effect of feeding pigs different dietary lipids, 3-wk-old, weaned pigs were fed for 40 d on one of five diets, which differed only in the type of oil present (the oil contributed 5% by weight of the diet and the total fat content of the diets was 8% by weight). The oils used were soybean (control diet), high-oleic sunflower oil (HOSO), sunflower oil (SO), canola oil (CO), and fish oil (FO; rich in long-chain [n-3] polyunsaturared fatty acids). There were no significant differences in initial or final animal weights, weight gains, or health scores among the groups. There were no significant differences in the concentration of anti-Escherichia coli vaccine antibodies in the gut lumens of pigs fed the different diets. The fatty acid composition of the diet markedly affected the fatty acid composition of the plasma and of mononuclear cells (a mixture of lymphocytes, monocytes, and macrophages) prepared from the blood, lymph nodes, or thymus. The FO feeding resulted in a significant increase in the number of circulating granulocytes. The FO feeding significantly decreased the proportion of phagocytes engaged in uptake of E. coli and decreased the activity of those phagocytes that were active. The proliferation of lymphocytes in cultures of whole blood from pigs fed the HOSO, SO, or FO diets was less than in those from pigs fed the CO diet. Proliferation of lymph node lymphocytes from SO- or FO-fed pigs was less than that from control, CO-, or HOSO-fed pigs. The natural killer cell activity of blood lymphocytes from pigs fed the FO diet was significantly reduced compared with those from pigs fed the CO diet. The concentration of PGE2 in the medium of cultured blood, lymph node, or thymic mononuclear cells was lower if the cells came from pigs fed the FO diet. Thus, the type of oil included in the diet of growing pigs affects the numbers and functional activities of immune cells in different body compartments.

Silicoating in resin-metal bonding: restorative implications for osseointegrated implant-bone unison.

Detailed background information leading to the development of a silicoating technique is presented with its advantages and disadvantages and its role in implant dentistry. The rationale is shown for coining a new acronym "osseointegrated Implant-Bone Unison" with an interest in biomechanical considerations for a prosthodontic restorative scenario. Several clinical cases are presented that involve silicoating and composite restorations.

Parotid and neck metastases from cutaneous squamous cell carcinoma of the head and neck.

BACKGROUND: Approximately 5% of cutaneous squamous cell carcinomas (SCC) metastasize to regional nodes. Nodal involvement may be more common for cutaneous squamous cell carcinomas of the head and neck, but few large series have reported clinical outcome after treatment of parotid gland and neck metastases from these cancers. PATIENTS AND METHODS: A combined retrospective/prospective study of patients treated between 1983 and 1994 was performed. There were 69 men and 6 women, with a median age of 67 years. Twenty-nine patients had neck metastases, and 33 had parotid gland metastases, while 13 patients had involvement at both sites. Of the 75 patients, 68 were treated surgically and 50 received postoperative radiotherapy. RESULTS: The facial nerve was sacrificed totally in 6 patients and partially in 9. Histologic extranodal spread was present in 48 (71%) of all surgically treated patients. Among 61 patients followed up to recurrence, or for greater than 12 months, 26 (43%) developed recurrence--12 in the parotid gland, 7 in the neck, and 7 in both sites. Multiple recurrences were common and occurred at a median of 8 months after surgery. Positive surgical margins were associated with poorer local disease control (P < 0.05). Cumulative survival at 5 years was 61%, but only 15 of 70 evaluable patients (21%) were eligible for follow-up at this time. Neck involvement with or without parotid gland disease was associated with an increased risk of distant metastases, but this was not statistically significant. Postoperative radiotherapy was not associated with improved disease control. CONCLUSION: Cutaneous, metastatic SCC involving the parotid gland and neck is an aggressive disease with a tendency to an infiltrative growth pattern and multiple recurrences. More aggressive surgery may be justified to reduce the incidence of regional failure after parotidectomy and neck dissection.

A soldering index made with 4-META adhesive resin.

Soldering indexes are most often made of fast-setting impression plaster or autopolymerizing resin. Lack of adhesive bonding between these materials and the casting requires the use of a bulky index to support the fixed partial denture components. The advent of adhesive resins improves the quality of the index and simplifies the procedure. A procedure for making a soldering index with 4-META adhesive resin is described that involves direct bonding of the resin to the metal castings. This procedure is accurate, simple, and time-saving. It can be used inside or outside the mouth, in prosthodontic or implant dentistry.