Everyday technology use among people with Parkinson's disease.

OBJECTIVES: To explore the relevance of and ability to use everyday technology (ET) among people with Parkinson's Disease (PD) and to explore associations between ET use and global cognition and motor ability. MATERIALS AND METHODS: Cross-sectional data was collected from 34 people with PD using the Short Everyday Technology Use Questionnaire+ (S-ETUQ+), the Movement Disorder Society-Unified Parkinson's Disease Rating Scale and the Montreal Cognitive Assessment (MoCA). RESULTS: Out of 41 ETs in the S-ETUQ+, the mean number perceived as relevant was 27.5 (min-max 19-35, SD 3.6). A good ability to use ET was reported where many ETs had a challenge measure below participants' ability to use them. A strong positive correlation between the ability to use ET and global cognition (MoCA) (r = .676, p = <0.01) was shown. CONCLUSIONS: ET use has become integrated into everyday life and is important for participation. This study showed a high relevance of and good ability to use ET and a correlation between ET use and global cognition among people with mild-moderate PD. Evaluation and support to use ET in PD are important for maintaining independence and participation, especially among those with cognitive decline.

Ultrasmall iron oxide particle contrast agent and MRI can be used to monitor the effect of anti-rejection treatment.

BACKGROUND: The purpose of the study was to investigate the feasibility of monitoring anti-rejection treatment using a blood pool contrast agent and magnetic resonance (MR) imaging. METHODS: Allogeneic heterotopic heart transplantations in rats were performed. In one group (treated group), a mild acute rejection was developed and subsequently treated and MR imaging was performed before and after anti-rejection treatment. In the other group (nontreated group), a mild acute rejection was developed and allowed to progress without treatment and MR examinations were performed before and after the advance of the acute rejection. After injecting ultrasmall superparamagnetic contrast agent, the relative change of signal intensity (SI) over time was measured. The SI difference between both radiological investigations for every animal was calculated; hence, every animal served as its own control. RESULTS: In both treated and nontreated groups, a significant difference over time was found between the two MR examinations seen as a decrease in the treated group and an increase in the nontreatment group. CONCLUSION: It is concluded that the effect of anti-rejection treatment can be detected using a blood pool agent and MR imaging, as a change in SI corresponding to changes in the vascular permeability.

T-cell inhibition does not aggravate bacterial translocation from rat small bowel.

BACKGROUND: T-cell mediated immunity has been proposed to have an important function in the defence against translocating microbes from the gastrointestinal tract. After small bowel transplantation massive T-cell immunosuppression is necessary to avoid rejection. As a consequence, infections with intestinal bacteria are the main contributors to mortality in this setting. This could further imply that T cells are important in limiting bacterial translocation. In a model for bacterial translocation from small bowel in the rat we examined the outcome of T-cell inactivation. METHODS: The studies were performed in a model of bacterial translocation from a Thiry-Vella loop of small bowel in the rat. The animals were treated with an anti-alpha/beta T-cell receptor monoclonal antibody (R73). Inhibition of T-cell activation was also made using the immunosuppressive drug cyclosporin A. All animals were sacrificed on day 3 postoperatively and translocation to the mesenteric lymph nodes, liver, spleen, lung and blood was evaluated. RESULTS: Treatment with R73 resulted in an almost complete labelling of T cells but did not result in any increased bacterial translocation compared to animals treated with saline. Neither did immunosuppression with cyclosporin A. CONCLUSIONS: In the model of bacterial translocation from a defunctionalised loop of small bowel the inhibition of T cells does not increase bacterial translocation to mesenteric lymph nodes or promote the systemic spread of the translocating bacteria. This indicates that T cells do not have any important protective function against translocating microbes from defunctionalised small bowel.

Phenotyping of ex vivo propagated graft-infiltrating cells-a tool to monitor rejection in the early post-operative period.

Objective and fast methods to diagnose rejection after organ transplantation are needed. In the present study, the ex vivo propagation technique was evaluated for its ability to detect rejection at two different time-points after experimental heart transplantation. Syngeneic and allogeneic heterotopic heart transplantations were performed using inbred rat strains. After 6 or 15 days, cardiac graft biopsies were put in culture and infiltrating cells isolated by the ex vivo propagation technique. The isolated cells were counted and phenotyped by flow cytometry. In parallel, graft sections were analysed with regard to morphology and the presence of infiltrating cells as determined by immunohistochemical stainings. On day 15 after transplantation, the number of cells possible to isolate through ex vivo propagation reflected the morphological changes of the graft, i.e. considerably more cells were obtained from allogeneic transplants undergoing rejection (1052 +/- 205) than from allogeneic grafts under cyclosporine protection (513 +/- 135; p < 0.05) or from syngeneic grafts (378 +/- 87; p < 0.01). Six days after transplantation the allogeneic grafts were strongly rejected with massive cellular infiltration, still there was no difference between allogeneic and syngeneic grafts as to the number of ex vivo propagated cells. However, the proportion of IL-2-receptor expressing T lymphocytes was increased (15.4 +/- 1.8% vs. 9.5 +/- 1.4%; p < 0.05) and the CD4/CD8 ratio reduced (1.0 +/- 0.1 vs. 2.8 +/- 0.2; p < 0.001) in the allogeneic group as compared with the syngeneic. We conclude that the ex vivo propagation technique can be used to distinguish rejection from non-rejection both early and later after transplantation, provided that not just cell counting but also phenotyping of the graft-infiltrating cells is performed.

Lymphocyte propagation from biopsies of kidney allografts.

Morphological evaluation of transplant biopsies, usually using the Banff classification, is the most important tool to diagnose rejection after kidney transplantation. However, morphological analysis only scores the amount and localisation of infiltrating cells, and studies show that up to 30% of grafts with a stable function display infiltration of lymphocytes consistent with acute cellular rejection. Methods to study the functional properties of the infiltrating lymphocytes are therefore needed. We applied a tissue culture system on biopsies from transplanted human kidneys, allowing infiltrating cells to propagate out from the tissue. Cells were then counted and subtyped by flow cytometry. The results were correlated to morphology. In total, 92 biopsies from 69 patients were analysed. For 14 patients, serial biopsies were available. In grafts with cellular or combined cellular and vascular rejection, the number of ex vivo propagated mononuclear cells was higher than from non-rejecting grafts. A similar pattern was seen for CD3(+) T cells as well as for T cells expressing CD25 or MHC class II antigens. However, the proportion of CD25(+) or MHC class II(+) T lymphocytes was similar in all groups (no rejection, vascular rejection, borderline changes, cellular rejection, combined cellular and vascular rejection). In all groups the number of CD4(+) cells was higher than the number of CD8(+) cells. The results confirm previous experimental studies showing that graft-infiltrating cells are possible to culture in vitro and that lymphocyte propagation correlates to acute cellular rejection. Tissue culturing is easy to perform and evaluate and can be used to determine and analyse the cellular immune response to allografts and may thus be used as a complement to morphological analyses.

Impact of consumption of freshwater fish on mercury levels in hair, blood, urine, and alveolar air.

Human exposure to methylmercury occurs mainly via consumption of fish. The aim of the study was to investigate the influence of freshwater fish consumption on mercury levels in hair, blood, urine, and end-exhaled air. Twenty subjects without dental amalgam fillings were recruited from sport-fishing societies. They ranged in age from 61 to 87 yr. Six individuals ate freshwater fish at least once a week and were categorized as high consumers. Eight individuals were classified as medium consumers and ate freshwater fish at least once a month but less than once a week. Six individuals were categorized as low consumers and had not eaten freshwater fish in the past 3 mo. Among the high consumers, median concentrations of mercury were 8.6 microg/L in blood, 2.4 microg/g in hair, 10 pg/L in end-exhaled air, and 1.1 microg/g creatinine in urine. The relationship between freshwater fish consumption and mercury was significant in all biological media. The high-consumption group had much higher mercury levels in blood (9-fold), hair (7-fold), alveolar air (3-fold), and urine (15-fold) than the low-consumption group. The latter finding may be explained by demethylation of methylmercury in the body. The ratio between mercury concentration in blood and hair was 1:270. This implies that the typical blood-hair ratio of 1:250, specified by the World Health Organization (WHO) in 1990, is valid also for exposure to low amounts of methylmercury.

Characterization of fibroblasts from rejecting tissue: the hyaluronan production is increased.

BACKGROUND: Interstitial edema of rejecting tissue can be partly attributed to the local accumulation of hyaluronan, which has strong water-binding capacity. The aim of the present study was to isolate fibroblasts from rejecting tissue and compare them, in terms of hyaluronan production and proliferation rate, with fibroblasts obtained from nontransplanted tissue. Furthermore, the fibroblast response to various cytokines involved in the rejection process was studied. METHODS: Fibroblasts were isolated from normal rat heart tissue and from cardiac allografts undergoing rejection. The various preparations were characterized with regard to hyaluronan production (radiometric assay) and cell proliferation ( H-thymidine incorporation). In addition, the effects of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and interleukin (IL)-2 on these parameters were studied. RESULTS: Fibroblasts isolated from rejecting hearts displayed strongly up-regulated hyaluronan production and proliferation rate as compared with fibroblasts obtained from normal tissue. In the presence of TNF-alpha, the proliferation of nonconfluent cells was augmented, whereas in confluent cultures of fibroblasts from rejecting tissue, the proliferation was inhibited. IFN-gamma stimulated both hyaluronan secretion and proliferation in confluent fibroblasts from rejecting hearts but had no effect on fibroblasts from normal tissue. IL-2, finally, reduced the hyaluronan production of nonconfluent cells. CONCLUSIONS: The activation of fibroblasts is increased in rejecting tissue. As a result, the hyaluronan concentration is elevated which, in vivo, contributes to the formation of an interstitial edema and a subsequently increased tissue pressure. Several cytokines present at rejection are involved also in the regulation of fibroblast activity.

Oral administration of xenogeneic erythrocytes induces production of antibodies that are capable of inducing hyperacute rejection of concordant vascularized xenografts.

Oral tolerance induction has proven to be an effective approach for inducing antigen-specific unresponsiveness in several models for allogeneic transplantation and autoimmune diseases. The authors' preliminary studies, however, indicated that xenospecific antibodies are produced when rats are given mouse erythrocytes orally. This response was further examined. Mouse erythrocytes were administered to rats orally or intravenously during one or two episodes, and sera were obtained on day 9 or day 29, respectively. Rat sera containing a positive hemagglutinating titer against mouse antigens were injected into rats that had recently undergone xenotransplantation to study graft survival. Oral administration of xenogeneic cells induced a powerful antibody response consisting mainly of xenospecific immunoglobulin (Ig) M and IgG. This antibody response also induced hyperacute rejection as powerfully as sera from intravenously immunized rats. The authors' study thus indicates that oral administration of xenogeneic cells is a powerful immunization pathway that induces an antibody response capable of rejecting concordant vascularized xenografts.

Quantification of lymphocytes propagating from rat-kidney allografts--a tool to monitor anti-rejection treatment.

BACKGROUND: Morphological evaluation of kidney biopsies applying the Banff Classification is closely correlated to clinical parameters of rejection. However, this classification is insufficient for monitoring the effect of instituted anti-rejection therapy. The objective of the present study was to develop a diagnostic tool enabling rapid assessment of the effects of an attempted anti-rejection treatment. MATERIALS AND METHODS: A kidney allotransplantation model in the rat was applied. All animals were initially maintained on cyclosporine A (CsA). In order to induce an episode of acute cellular rejection, CsA was withdrawn for 5 days. Thereafter, the immunosuppressive treatment was restarted. Core biopsies were taken before, and at various times after, the introduction of anti-rejection therapy. Infiltrating cells were isolated using an in vitro culture system, allowing cells to propagate from the biopsies to culture medium. Propagated cells were counted and analysed for subtype and activation markers using flow cytometry. The results were compared with immunohistochemical and morphological analyses of the grafts. RESULTS: By applying the in vitro culture system it was possible to demonstrate a reduction in outgrowth of mononuclear cells from core biopsies as early as 2 days after the start of anti-rejection therapy. At this time-point, as well as 2 days later, morphological and immunohistochemical analyses of biopsies showed ongoing acute cellular rejection, with no differences between biopsies taken before and after restart of CsA-therapy. The percentage of propagating T lymphocytes expressing the activation markers CD25 or MHC class II did not differ between grafts with ongoing rejection and grafts obtained from CsA-treated rats, neither did the ratio of CD4/CD8-positive cells. CONCLUSION: Our findings indicate that the ex vivo propagation method could function as a complement to routine histology, not only in the diagnosis of cellular rejection but also in order to rapidly reveal a failure of an anti-rejection therapy to halt the rejection process.

COX-2 inhibitors prolong trauma-induced elevations of iris hyaluronan.

PURPOSE: To investigate whether and how treatment with COX-2 inhibitors influences hyaluronan responses to a standardized trauma, argon laser induced iritis, in rabbits. METHODS: Two different COX-2 inhibitors were used, SC-236 and rofecoxib. The drugs were administered orally, 6 mg/kg/day and 1.5 mg/kg/day respectively. Iris and aqueous humor hyaluronan concentrations were measured with a radiometric assay at different time points after laser irradiation. RESULTS: The hyaluronan concentration in the iris increased 3-4-fold with a peak concentration of 129.1 microg/g wet weight 2 days after laser irradiation. It then decreased to normal values after 1 week. In eyes treated with either of the COX-2 inhibitors, iris hyaluronan concentrations did not decrease as rapidly and were significantly higher at day 4 and 7 when compared to drug untreated eyes. CONCLUSION: Treatment with COX-2 inhibitors prolongs trauma induced elevation of iris content of endogenous hyaluronan. This may be, at least partly, due to an inhibition of interstitial fluid pressure regulation.

Hyaluronan synthases and hyaluronidases in the kidney during changes in hydration status.

Hyaluronan is a large glycosaminoglycan that is abundant in the interstitium of the renal medulla/papilla. Papillary hyaluronan increases during hydration and decreases during dehydration. Due to its gel properties and ability to retain large volumes of water, hyaluronan plays a role in renal water handling by affecting the permeability characteristics of the papillary interstitium. The focus of the present investigation was the regulation of hyaluronan metabolism in the kidney, especially during variations in hydration status. In control papillas, HAS 2 mRNA was heavily expressed and HAS 1 and 3 mRNA were weakly distributed. HYALs 1-3 mRNA were found at high expression and HYAL 4 was only weakly expressed. In hydrated animals, the diuretic response (12-fold) was followed by a 58% elevation in papillary hyaluronan and a 45% reduction in the excreted urinary hyaluronidase activity. No difference was determined in HAS 1-3 mRNA or HYAL 1, 3-4 mRNA expression, suggesting a change in activity rather than amount of protein. In dehydrated animals, antidiuresis was followed by a 22% reduction in papillary hyaluronan and a 62% elevation in excreted urinary hyaluronidase activity. Plasma vasopressin was 2.8-fold higher in dehydrated vs. hydrated rats. In conclusion, HAS 2 appears a major contributor to the baseline levels of hyaluronan. Reduced HAS 2 gene expression and increased excreted urinary hyaluronidase activity during dehydration contribute to the reduced amount of hyaluronan and to antidiuretic response.

Serum hyaluronan--a potential marker of cardiac allograft rejection?

BACKGROUND: The connective tissue component hyaluronan accumulates within the transplanted organ at rejection. Increased tissue content of hyaluronan is seen also in synovial tissue in rheumatoid arthritis and in skin in scleroderma. In these diseases, the elevated hyaluronan levels are reflected by increased concentrations of hyaluronan in serum. The aim of the present study was to study the changes in serum hyaluronan after organ transplantation. METHODS: The experiments were performed in a rat model of heterotopic heart transplantation. Serum hyaluronan was assessed at various times after allogeneic (rejection) and syngeneic (non-rejection) transplantation and correlated with tissue hyaluronan. In addition, serum hyaluronan in animals who had long-term-surviving allogeneic grafts was studied. RESULTS: The hyaluronan concentration in serum was significantly higher in the rejecting than in the non-rejecting group 4 and 6 days after transplantation (p < 0.01). On Day 6, serum hyaluronan had increased by 400% in animals with an allogeneic transplant and by 100% in those with a syngeneic transplant. There was a positive correlation between serum hyaluronan and tissue hyaluronan (p < 0.05). Animals with long-term-surviving grafts displayed normal serum hyaluronan levels. CONCLUSIONS: Rejection of rat heart transplants is associated with strongly increased serum hyaluronan that parallels the hyaluronan accumulation within the transplant.

Regulation of fibroblasts by activated and non-activated immune cells.

BACKGROUND: Rejection of transplanted tissue is characterized by cell infiltration and interstitial edema. Graft fibroblasts and fibroblast products are partly involved in the regulation of both these phenomena. Knowledge about the mechanisms behind fibroblast activation may lead to new strategies to prevent rejection. This study investigated whether cells of the immune system have the capacity to regulate fibroblast activation. METHODS: Fibroblasts isolated from rejecting heart transplants or from normal heart tissue were cultured in the presence of supernatants of stimulated or non-stimulated immune cells. The immune cells were challenged either in vitro (incubation with phytohemagglutinin) or in vivo (organ transplantation). Fibroblast proliferation and hyaluronan production were measured. RESULTS: Normal, sub-confluent heart fibroblasts showed an increased proliferation rate in the presence of supernatants of activated immunocompetent cells, irrespective of if these cells had been stimulated in vitro or in vivo. As expected, proliferation rate and hyaluronan production were upregulated in fibroblasts isolated from rejecting tissue. However, supernatants of biopsy specimens obtained from non-rejecting organs (syngeneic transplants or normal hearts) had an inhibitory effect on the growth rate of confluent fibroblasts isolated from rejecting tissue. CONCLUSIONS: We conclude that graft-infiltrating cells and immune cells activated in vitro have the capacity to stimulate fibroblasts, most probably as a result of the production and secretion of fibroblast-stimulating factors.

The graft content of hyaluronan is increased during xenograft rejection.

BACKGROUND: Hyaluronan, a macromolecule with strong water binding capacity, is associated with interstitial oedema during rejection of allogeneic transplants. However, the involvement of hyaluronan during xenograft rejection has previously not been investigated. The aims of this study were to characterize hyaluronan content and distribution during rejection of concordant mouse-to-rat cardiac xenografts, and to explore the effects of hyaluronidase (HAse) on xenograft survival. METHODS: Graft recipients were treated with 15-deoxyspergualin (DSG) or both HAse and DSG. Grafts were removed on day 5 from some of the animals to analyse hyaluronan and water content, while other animals were used to investigate graft survival. The hyaluronan content was measured by a radiometric assay and the distribution was analysed by histochemical staining. RESULTS: In xenografts undergoing rejection (the DSG group) there was a strong increase of the hyaluronan [555 +/- 93 microg/g dry weight (dw)] and water (82.7 +/- 0.4%) contents compared with normal mouse heart tissue (166 +/- 10 microg/g dw; P < 0.01 and 78.6 +/- 0.5%; P < 0.001, respectively). The combined use of HAse and DSG reduced the accumulation of hyaluronan (284 +/- 43 microg/g dw; P < 0.05 vs. DSG) but did not affect the average water content. The average graft survival time did not differ between the groups; however, three grafts in the HAse + DSG-treatment group survived much longer than the longest-surviving grafts in the DSG group. CONCLUSIONS: These data suggest that the graft content of hyaluronan considerably increases during xenograft rejection. HAse effectively reduces this accumulation, but does not affect the average water content.

Isolation of mouse-to-rat cardiac xenograft-infiltrating cells by ex vivo propagation.

Ex vivo propagation of graft infiltrating lymphocytes has become a useful method for the examination of the cellular response after allogeneic transplantation. The aim of the present study was to investigate if this method can be used also for isolation of xenograft infiltrating cells, and, if so, to further characterize these cells. The concordant mouse-to-rat heart transplantation model was used for the experiments. Recipient rats were treated either with 15-deoxyspergualin (DSG) or with a combination of DSG and cyclosporine A (CyA) or left untreated. Transplants were removed beating on day 2 (untreated) or day 8 (DSG and CyA + DSG) and biopsies were incubated in culture medium for 48 h, resulting in propagation of cells from the biopsies into culture medium. The propagated cells were counted and phenotyped using flow cytometry. In parallel, the transplants were examined morphologically and immunohistochemically. Infiltrating cells could be isolated from all grafts in all groups. The number of propagated T lymphocytes during cellular rejection (DSG-treatment) was about 3.5 times higher than during 'non-rejection' (CyA + DSG) and six times higher than during acute vascular rejection (untreated). These findings were supported both by the morphological and the immunohistochemical findings. In conclusion, we have shown that graft infiltrating lymphocytes can be isolated from xenogeneic heart transplants by incubation of biopsies for 48 h, resulting in spontaneous propagation of immune cells into culture medium. Propagated cells could be further characterized by flow cytometry. Thus, the technique presented here can be used as a tool for studies of xenogeneic cellular rejection.

Renomedullary and intestinal hyaluronan content during body water excess: a study in rats and gerbils.

Our previous studies in rats have suggested a role for renomedullary hyaluronan (HA) in water homeostasis. The gerbil is known for its unique ability to conserve water. In the present study renal papillary and intestinal HA were compared between groups of anaesthetized gerbils and rats before and after up to 6 h of I.V. water loading. Baseline papillary HA in gerbils was only 37 % of that in the rat. Water loading in rats increased the papillary HA content. Elevation was maximal (+27 %, P < 0.05) after 2 h of water loading and then declined to control levels after 6 h of water loading (+3 %, n.s.). In contrast, the gerbil responded with a decreased papillary HA content during water loading. The depression was maximal after 2 h (-49 %, P < 0.05) and was still 41 % below the control values after 6 h (P < 0.05). The urine flow rate increased rapidly in the rat and its maximum, 21 times above the control level (P < 0.05), occurred at the HA peak, i.e. after 2 h of water loading while in the gerbil, the urine flow rate increased slowly and slightly and was only six times above control values after 6 h of water loading (P < 0.05). The HA content along the intestine was similar in the two species: lowest in the duodenum and jejunum and highest in the distal colon. To conclude, in the rat, the elevation of papillary interstitial HA during acute water loading would counteract water reabsorption by changing the physico-chemical characteristics of the interstitial matrix favouring rapid water diuresis. This would work as a complement to the powerful regulation by ADH. The gerbil has a diametrically different regulation of papillary HA turnover during water loading. The decreased papillary HA level during water loading and the slow and small diuretic response may represent a genetic difference in adaptation to enhance the ability to conserve water in an arid environment.

Hair mercury levels versus freshwater fish consumption in household members of Swedish angling societies.

Hair mercury levels were determined in 143 individuals from households of members in angling societies in an area of Sweden with many lakes that have freshwater fish with relatively high mercury levels. Thus, the individuals had a potentially high intake of methyl mercury. The mean mercury concentration of pike and perch was approximately 0.7 microg/g. One-third of the subjects consumed these freshwater fish at least once a week. As could be expected, there was a clear increase in hair Hg with reported freshwater fish consumption (P < 0.001). The median mercury level in hair was 0.9 microg Hg/g for the whole group, and for those who reported consumption of freshwater fish at least once a week it was 1.8 microg/g. The highest hair mercury level was 18.5 microg/g, in a man who consumed pike and perch several times per week. Men had higher hair Hg than women, also when stratified for fish consumption. This was verified in 32 couples, of which the man and woman consumed the same fish and reported the same consumption. The median hair mercury level in these 32 couples was 1.3 microg/g for men and 0.8 microg/g for women (P = 0.002). About half of the subjects had hair mercury exceeding 1 microg/g, corresponding to the reference dose (RfD) of 0.1 microg of mercury per kilogram body weight set by the US Environmental Protection Agency. Although the RfD applies to all populations, the most at-risk group at these levels is pregnant women. There were only 2 women (of 12) of fertile age with hair mercury above 1 microg/g. In Sweden pregnant women are advised not to eat perch and pike at all during pregnancy. Since fish is rich in many important nutrients, it is unsatisfactory that fish consumption must be restricted, and thus there is a need to reduce mercury levels in fish.

Effects of increased intra-abdominal pressure and volume expansion on renal function in the rat.

BACKGROUND: The effects of increased intra-abdominal pressure (IAP) and volume expansion on renal function in the rat were studied to gain more knowledge of the oliguria seen during laparoscopic procedures and to reduce the detrimental renal effects of IAP. METHODS: IAP was elevated to 5 or 10 mmHg by insufflation of CO(2) and maintained for 2 h in anaesthetized and mechanically ventilated rats. Rats with normal IAP served as controls. An angiotensin II receptor I antagonist, candesartan, was given as a bolus injection and a 5% volume expansion was achieved by i.v. saline infusion. An angiotensin-converting enzyme (ACE) inhibitor was also given. Renal parameters were the glomerular filtration rate (GFR), urine production, the urinary concentrations of sodium and potassium and the osmolality in the urine. The arterial acid-base balance and blood pressure were also monitored. RESULTS: The GFR deteriorated by 70% during pneumoperitoneum (PP) of 10 mmHg. There was a dramatic drop in sodium excretion (88-97%). With candesartan and elevated IAP, there was a drop in mean arterial pressure (from 90 to 55 mmHg) and the negative renal effects were very pronounced. Renal function was better preserved during elevated IAP in combination with volume expansion. CONCLUSIONS: Capnoperitoneum suppresses renal function, especially in combination with angiotensin II receptor 1 blockade and ACE inhibition. Volume expansion reduces the deleterious effects of PP on renal function during elevated IAP. The results suggest that patients should not be given pharmaceuticals blocking the renin-angiotensin-aldosterone system prior to procedures that may increase IAP. It may be beneficial, however, to reduce angiotensin II tension by volume expansion.

Renal hyaluronan accumulation and hyaluronan synthase expression after ischaemia-reperfusion injury in the rat.

BACKGROUND: Hyaluronan (HA) is a connective tissue component with unique water binding and pro-inflammatory properties. It has been suggested that HA is involved in normal renal water handling but also in several pathological conditions such as organ rejection and ischaemia-reperfusion (IR) injury. METHODS: In anaesthetized normal rats we investigated if renal cortical HA accumulation and the intrarenal distribution and expression of HA synthases (Has 1, 2 and 3) correlate with renal dysfunction after renal IR injury. After 20, 30 or 45 min of unilateral renal ischaemia and 72 h of reperfusion, renal function and cortical HA content were measured. Has 1, 2 and 3 mRNA were determined in control and IR kidneys subjected to 45 min ischaemia and 72 h reperfusion. RESULTS: IR kidneys had reduced urine concentrating ability, potassium excretion, glomerular filtration rate (GFR) and renal blood flow. On average, IR kidneys had more than 10 times higher amounts of cortical HA than the contralateral control kidney and their water content was elevated while medullary HA was largely unaffected. Has 2 expression in the cortex was heavily up-regulated in IR kidneys while Has 3 remained at control levels. Has 1 could never be detected. There was a direct correlation between the amount of cortical HA and the time period of ischaemia and also between the cortical amount of HA and depression of functional parameters. CONCLUSIONS: IR injury depresses parameters of renal function, which coincides with an elevated cortical HA content and Has 2 expression. The enhanced Has 2 expression indicates that the cortical HA accumulation is primarily dependent on increased HA synthesis and not impaired degradation/elimination. The water binding and pro-inflammatory properties of HA may contribute to renal dysfunction after IR.

Intragraft cytokine mRNA expression in rejecting and non-rejecting vascularized xenografts.

BACKGROUND: The aim of the present study was to further investigate the characteristics of both graft-infiltrating cells and splenocytes during acute vascular rejection (AVR), cell-mediated rejection and non-rejection of vascularized concordant xenografts, by analysing both proinflammatory [interleukin-1beta (IL-1beta) and tumour necrosis factor (TNF-alpha)] and more specific [(IL-2, IL-4, IL-10, IL-12p40 and interferon-gamma (IFN-gamma)] cytokines. A parallel investigation was made of the antibody response of IgM and IgG to the xenografts. METHODS: Mouse hearts were heterotopically transplanted to the neck vessels of recipient rats. Grafts, spleens and sera were collected from untreated (AVR) and cyclosporin A (CyA) treated animals on day 2 after transplantation. Organs from rats treated with 15-deoxyspergualin (DSG) or CyA and DSG in combination were harvested on both day 2 and day 8. Grafts from DSG-treated rats undergo cell-mediated rejection and stop beating on day 9 and forth, while CyA + DSG treatment results in long-term graft survival. Real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was applied for analysis of intragraft and splenic cytokine messenger RNA (mRNA) expression. The phenotypes of the graft infiltrating cells were characterized by immunohistochemistry. The antibody response was investigated by means of immunofluorescence, haemagglutination and flow cytometry. RESULTS: All the studied cytokines (IL-1beta, IL-2, IL-4, IL-10, IL-12p40, IFN-gamma and TNF-alpha) were up-regulated in the grafts from rejecting untreated (day 2) and DSG-treated animals (day 8) in comparison with grafts from CyA + DSG treated animals (day 8). On day 2 under immunosuppression with CyA, DSG or CyA + DSG no or low cytokine mRNA levels were found. The mRNA levels of IL-2, IL-4 and IFN-gamma in the spleens were suppressed under both DSG- and CyA + DSG treatment on day 8. Immunofluorescence showed deposits of both IgM and IgG in grafts from untreated, CyA-treated (day 2) and DSG-treated (day 8) animals, while CyA + DSG treatment diminished these deposits on both day 2 and day 8. No circulating antibodies were identified in either group. CONCLUSION: We hereby conclude that both AVR on day 2 and cell-mediated rejection on day 8 (under DSG treatment) in a concordant cardiac mouse-to-rat xenotransplantation model are associated with an increase of proinflammatory cytokines, T helper 1 (Th1)-associated cytokines as well as IL-10, while immunosuppression with CyA + DSG diminishes the levels of all examined cytokines. Grafts undergoing AVR or cellular rejection are subjected to deposits of both IgM and IgG, although circulating donor specific antibodies are undetectable in serum.