Tolerance induction ameliorates allograft vasculopathy in rat aortic transplants. Influence of Fas-mediated apoptosis.

Based on successful induction of donor-specific unresponsiveness by alloantigenic stimulation in several animal models of acute rejection, we hypothesized that similar immune manipulations would also inhibit the evolution of chronic rejection and transplant vasculopathy. To induce immune tolerance, DA rats received a PVG heart allograft and were immunosuppressed with cyclosporine for 30 d. At day 100 the animals were challenged with a PVG aortic allograft after either 1 or 18 h of cold ischemia. 8 wk after the aortic transplantation, the grafts were investigated for morphological changes, infiltrating cells, apoptosis, and Fas-Fas ligand expression. Control allografts showed advanced transplant arteriosclerosis, whereas tolerance-induced aortic allografts displayed reduced neointimal formation, less medial atrophy, fewer apoptotic cells, and fewer Fas- and FasL-expressing cells. Prolonged ischemic storage time did not profoundly alter the morphological changes of the allografts. Fas expression was found in T cells, macrophages, vascular smooth muscle cells, and endothelial cells, whereas FasL was expressed mainly by T cells and macrophages. FasL mRNA expression was evident throughout the entire allograft wall. In conclusion, induction of allospecific tolerance can effectively prevent transplant arteriosclerosis. Cold ischemia damage does not abrogate the beneficial effect of tolerance, but creates a separate identity of mainly endothelial lesions. Furthermore, Fas-mediated apoptosis appears to be involved in the pathological lesions seen in chronic rejection.

Renomedullary interstitial cells in culture; the osmolality and oxygen tension influence the extracellular amounts of hyaluronan and cellular expression of CD44.

Our previous studies have suggested a role for renomedullary interstitial cells (RMICs) and renal medullary hyaluronan (HA) in water homeostasis. In the present study, cultured rat RMICs were used to examine the relationship of osmolality and oxygen tension on the extracellular amount of HA in the culture and to the cellular immunoreactivity to CD44, a HA binding protein. Under isotonic (330 mOsm(.)kg(-1) H(2)O), normoxic (20% O(2)) conditions, supernatant from sub-confluent RMICs contained 120+/-37 pg 10(4) cells(-1) 24 h(-1) of HA. Under hyperosmotic conditions (630 mOsm kg(-1) H(2)O), HA in the supernatant was decreased by 42% and under hypoosmotic conditions (230 mOsm kg(-1) H(2)O) it was doubled. Under hypoxic, iso-osmolar conditions (5% and 1% O(2), 330 mOsm kg(-1) H(2)O) this HA content was decreased by 56 and 48%, respectively, compared with normoxic, iso-osmolal conditions. Expression of CD44 on sub-confluent cells increased with increasing osmolality, as shown by immunostaining and flow cytometric analysis. The increases in CD44 from 330 to 630, 930 and 1230 mOsm kg(-1) H(2)O amounted to 5, 142 and 212%, respectively. Low oxygen tension (5% O(2)) decreased the intensity of CD44 immunofluorescence by 31%. Cell viability was similar at all conditions studied. In summary, these data indicate that cultured RMICs produce HA and are immunoreactive to CD44. In the supernatant of RMICs, the HA content decreases under hyperosmotic, hypoxic conditions. Conversely, CD44 immunoreactivity increases under hyperosmotic conditions. These results may explain our previous in vivo findings of a decreased renal papillary HA content during anti-diuresis and an increased content during water diuresis. The results support the concept that RMICs play an important role in renal water handling.

Ventricular proliferation zones in the brain of an adult teleost fish and their relation to neuromeres and migration (secondary matrix) zones.

Zones containing actively dividing cells (proliferation zones: PZs), in the brain of adult three-spined sticklebacks, were identified by autoradiographic detection of (3)H-thymidine and immunocytochemical detection of the thymidine analogue 5'-bromodeoxyuridine (BrdU), singly or in combination, and by immunocytochemical detection of proliferating cell nuclear antigen (PCNA) by monoclonal antibodies. The PZs are associated with boundaries between adult brain regions, as well as with defined morphofunctional subdivisions. PZs are located at the border between the telencephalon and diencephalon, and at the border between the mesencephalon and the rhombencephalon. In the midbrain, the PZ follows the dorsomedial, caudal, and ventrolateral aspects of each tectal hemisphere, extending over the caudal aspect of the torus semicircularis to the nucleus lateralis valvulae. In the hindbrain, the major PZ apparently represents the persisting embryonic secondary matrix layer of the developing cerebellum. In the forebrain, the PZs are associated with the ventricular zones of the olfactory bulbs and ventral telencephalic area ("subpallium"), dorsal telencephalic area ("pallium"), preoptic region, ventral thalamus, dorsal thalamus, epithalamus, pretectum, posterior tuberculum, and the hypothalamus. The diencephalic PZs are parcellated according to a neuromeric organisation (a synencephalic, a posterior, and an anterior parencephalic neuromere: p1, p2, and p3). The PZs of the secondary prosencephalon (telencephalon and hypothalamus) thus would belong to neuromeres p4-6, but do not form an immediately recognised serial pattern. The prosencephalic PZs correlate well with parts of embryonic migration areas as defined by Bergquist and Källén ([1954] J. Comp. Neurol. 100:627-659), morphogenetic fields from which postmitotic neurones migrate to their final destination.

Monitoring of intragraft pressure of rejecting organs: increased tissue pressure can be reduced by hyaluronidase therapy.

BACKGROUND: The present study was undertaken in order to: (a) develop a new technique for measurement of interstitial pressure, (b) study the intragraft pressure of rejecting and non-rejecting organs, and (c) study the effect of treatment with the hyaluronan-degrading enzyme hyaluronidase on intragraft pressure. Treatment with hyaluronidase has previously been demonstrated to result not only in reduction of tissue hyaluronan but also in ameliorated interstitial edema, and we suggested that the diminished edema would lead to a reduced interstitial pressure as well. METHODS: At day 5 after syngeneic or allogeneic rat heterotopic heart transplantation, the interstitial pressure of the cardiac grafts was measured using a microtip pressure sensor. Subsequently, the allogeneically grafted animals received a continuous intravenous infusion of either hyaluronidase (total dose: 60,000 U/kg) or vehicle during 2 hr; meanwhile, the interstitial pressure was monitored. RESULTS: The intragraft pressure measurement technique was found to give reproducible results. The interstitial pressure of the rejecting (allogeneic) grafts was considerably higher than that of the non-rejecting (syngeneic), i.e., 12.3+/-1.6 mmHg vs. 1.1+/-0.6 mmHg (P<0.001). Hyaluronidase infusion effectively reduced the interstitial pressure as compared with vehicle treatment. By 20 min, the pressure had been reduced by 28% (P<0.01 compared with vehicle treatment); after 1 hr, by 49% (P<0.001); and after 2 hr, by 68% (P<0.01). CONCLUSIONS: By using modern technology for tissue pressure measurements, we found that the strongly increased interstitial pressure of rejecting organs can be instantly reduced by intravenous administration of the hyaluronan-degrading enzyme hyaluronidase.

Recipient-reactive antibodies occur during development of acute graft-versus-host reaction after small bowel transplantation.

After small bowel transplantation, not only rejection but also graft-versus-host reaction (GVHR) may occur. Donor T lymphocytes, transferred together with the graft, are a prerequisite for the development of GVHR. So far, however, little is known about the effector mechanisms in acute GVHR. It can be assumed that not only T lymphocytes but also other cells, i.e., B lymphocytes, monocytes/macrophages, and NK cells, together with inflammatory cytokines, are responsible for the lesions of recipient tissue. In the present study, the occurrence of recipient-reactive antibodies after semisyngeneic small bowel transplantation was investigated to elucidate the role of B lymphocytes in GVHR development. No antibodies reactive with recipient cells were detectable in serum from untreated, nontransplanted rats. Five days after transplantation, recipient-reactive antibodies started to appear in recipient serum. At the same time, a deposition of IgM antibodies became visible in the liver and native intestine, which are target organs for GVHR. No antibodies directed against either the donor strain or a third-party strain were detectable in serum. We conclude that a synthesis of antibodies against recipient tissue occurs during the development of GVHR. Whether these antibodies contribute to the disease remains unclear.

Recovery of hyaluronan during perfusion of small bowel transplantation reflects rejection.

Rejection seen after small bowel transplantation (SBT) in the rat is associated with an accumulation of hyaluronan (HA) in the lamina propria of the graft. In this study the intestinal intraluminal content of this connective tissue component was measured in order to determine if HA could be used as an index of rejection following SBT. Syngeneically and semiallogeneically transplanted rats were investigated by perfusion of a 4-cm segment of the intestinal graft on days 2, 4, and 6 posttransplantation. The amounts of HA recovered during perfusion were analyzed using a radiometric assay. In rejecting grafts the recovered HA amounts increased 15 times from day 2 to day 6, although after syngeneic transplantation there was only a minor increase from day 2 to day 4 and no further increase on day 6. Thus, the recovery of HA during perfusion of transplanted small bowel grafts has the potential value to reflect rejection.

Simultaneous occurrence of rejection and graft-versus-host reaction after allogeneic small bowel transplantation.

Small bowel transplantation may be complicated not only by rejection but also by graft-versus-host reaction (GVHR). So far, little is known about the association between these two immunological reactions, partly because of the lack of standardized, reproducible experimental models for such studies. In this work, a rat model in which GVHR and rejection occur simultaneously was established. When transplanting small bowel grafts from BN donors to Lewis recipients and thereafter treating the grafts locally with the immunomodulating substance LS-2616, a clinically visible GVHR occurred on day 6, at the same time the first signs of rejection became visible. The GVHR was confirmed by immunohistochemical staining for MHC class II-positive cells in liver and ear skin biopsy specimens. An obvious quantitative difference in the number of positive cells in both organs was observed when local treatment was compared with oral LS-2616 treatment or with findings in organs from untreated animals. We conclude that GVHR and rejection are not mutually exclusive and thus may occur simultaneously, and that this pharmacological model might facilitate further studies of the impact of GVHR on graft rejection and recipient survival.

Successful retransplantation of mouse-to-rat cardiac xenografts under immunosuppressive monotherapy with cyclosporine.

The present study was undertaken to investigate whether retransplantation with a second xenograft, from the same species as the primary graft, is possible to achieve using only moderate immunosuppression. Heterotopic mouse-to-rat cardiac transplantations were performed, and the recipients were treated with 15-deoxyspergualin (DSG) and cyclosporine (CsA) at high doses for days -1 to 4 and at moderate doses for days 5 to 28. From day 29 and onward, the immunosuppressive protocol consisted of daily oral administration of CsA 10 mg/kg as monotherapy. Animals that had beating grafts when DSG treatment was stopped were retransplanted 56-154 days after the primary transplantation, either with a vascularized graft (heart) or with nonvascularized graft (pancreatic islets), under continued therapy with CsA. Six of 10 secondary cardiac xenografts functioned for more than 50 days and were harvested beating after 60-100 days. In contrast, nonimmunosuppressed or DSG-treated rats are known to reject a second cardiac mouse graft hyperacutely. The unresponsiveness was confined to cardiac tissue, as the pancreatic islets, transplanted under the kidney capsule, were totally rejected after 14 days. Long-term functioning cardiac xenografts, primary and secondary, had a well-preserved morphology, and infiltrating mononuclear cells were found just in the periphery of the grafts. A majority of these cells were macrophages expressing the ED1, but not the ED2, antigen. No deposition of IgG or complement was seen in any of the graft vessels, whereas a slight deposition of IgM was observed in some vessels of both primary and secondary grafts. In conclusion, we have demonstrated that unresponsiveness can be induced by effective immunosuppression of the recipient at the time of the initial transplantation, so that retransplantation with a second xenograft can be performed successfully under single-drug immunosuppressive therapy with CsA.

Alterations in mucosal immune cell distribution in septic rats.

A reduced cell-mediated immunity in the intestinal mucosa might promote gut barrier failure and bacterial translocation in shock. This study was performed to investigate changes from the normal distribution of cellular components of the immune system within the intestine during sepsis. Intra-abdominal sepsis was induced by cecal ligation and puncture. After 24 h, the animals were killed and specimens were taken from the small bowel. Immunohistochemical stainings were performed on frozen sections using monoclonal antibodies reactive with MHC class II positive cells (OX6), the alpha/beta antigen receptor on T lymphocytes (R73), CD4+ T lymphocytes (W3/25), CD8+ T lymphocytes (OX8), and macrophages (ED1). There was a significant reduction in the number of pan T lymphocytes as well as both CD4+ and CD8+ subsets in the mucosa of the septic rats as compared with sham operated rats (p < .01). In contrast, the populations of MHC class II positive cells and macrophages increased in sepsis (p < .01). We conclude that 24 h after the induction of peritonitis, there is an altered pattern of immunocompetent cells within the intestinal mucosa.

Edema treatment during cardiac allograft rejection.

BACKGROUND: Interstitial edema of rejecting organs can be correlated with the accumulation of hyaluronan in the transplant; since hyaluronan has strong water-binding capacity, treatment with the hyaluronan-degrading enzyme hyaluronidase reduces not only the hyaluronan content but also the water content of the graft. The aim of the present study was to investigate whether a further reduction of the water content would be the result if hyaluronidase was used in conjunction with classic diuretic substances. METHODS: Five days after heterotopic heart transplantation (PVG to Wistar/Kyoto), recipient rats received hyaluronidase as a continuous intravenous infusion over 2 hours together with either a loop-diuretic (furosemide) or an osmotic diuretic (mannitol). RESULTS: Hyaluronidase was found to reduce the hyaluronan contents of the grafts from 586+/-52 microg/g in control animals receiving vehicle infusion to 161+/-48 microg/g (p < 0.001) and the water contents from 81.3+/-0.4 x 10(-2) U to 79.7+/-0.4 x 10(-2) U (p < 0.05). Combined treatment with furosemide or mannitol did not affect the results and neither furosemide nor mannitol had any intrinsic capacity to affect the water or hyaluronan contents of the cardiac grafts. CONCLUSION: This experiment confirms our previous findings of hyaluronidase as an effective edema-reducing drug and indicate that no additive effect is obtained by a combined therapy with diuretics and hyaluronidase.

The immunomodulator LS-2616 (linomide) more effectively than sensitization of the donor enhances graft-versus-host reaction after small bowel transplantation.

The effects of the immunomodulating substance LS-2616 (linomide) on graft-versus-host reaction (GVHR) were investigated in a semi-syngeneic small bowel transplantation model. The entire bowel of Lewis donors were transplanted heterotopically into (Lewis x BN) F1 hybrids. Both untreated animals and animals treated with LS-2616, in a daily dose of 160 mg/kg, developed a lethal GVHR. The median survival time in untreated animals was 14.5 days while in LS-2616 treated animals it was just eight days (p < 0.01). LS-2616 in combination with cyclosporin A (CyA), 15 mg/kg given orally on days 0-20, did not seem to alter the survival times compared with CyA treatment alone; 56% of the animals treated with CyA survived for more than 100 days and after combined treatment with CyA/LS-2616 there were 50% permanent survivors. Also the effect of earlier sensitization of the donor on the course of GVHR was investigated. Hearts from BN rats were transplanted heterotopically to the neck vessels of Lewis rats. The hearts were rejected on about day six; five days later the bowels were harvested and transplanted into (Lewis x BN) F1 hybrids. The median survival time in this group was 12.5 days. Taken together our results, in combination with earlier findings, suggest that, at the level of effector mechanisms, GVHR is not an exact mirror image of rejection. Also, LS-2616 appears to be a useful tool for further studies of the mechanisms of action of GVHR.

Role of hyaluronan in acute pancreatitis.

BACKGROUND: The connective tissue component hyaluronan is accumulated locally in the damaged tissue during various inflammatory conditions. Owing to the strong water-binding capacity of this glycosaminoglycan, increased tissue content of hyaluronan is paralleled by the development of interstitial edema. The aim with the current experiment was to investigate whether hyaluronan is accumulated in acute pancreatitis and if increased levels of hyaluronan can be correlated to the inflammation of the pancreatic tissue. METHODS: Acute pancreatitis was induced in Sprague-Dawley rats by the administration of supramaximal doses of the cholecystokinin analogue caerulein. The animals were followed for 5 hours (n = 4), 24 hours (n = 6), or 48 hours (n = 5), and the pancreata were then investigated for hyaluronan and water content, hyaluronan distribution, general morphology and the presence of CD44-positive cells, macrophages, and T lymphocytes. RESULTS: Hyaluronan accumulated in the edematous interstitium during acute pancreatitis. Twenty-four hours after the induction of pancreatitis, the hyaluronan content of the pancreata had increased by more than 100%. Simultaneously, CD44-positive cells infiltrated the tissue. However, no correlation between hyaluronan and water was seen at any time point. CONCLUSIONS: This study shows that acute pancreatitis is associated with a strong but transient increase in interstitial hyaluronan and an infiltration of CD44-positive cells located mainly in the same region as the accumulated hyaluronan.

The effects of combined treatment with the novel vitamin D analogue MC 1288 and cyclosporine A on cardiac allograft survival.

The efficacy of combined treatment with the novel vitamin D analogue MC 1288 (20-epi-1 alpha,25-dihydroxycholecalciferol) and cyclosporine A (CyA) in preventing rejection following organ transplantation was evaluated in the heterotopic cardiac allograft model. Wistar/Kyoto rats received hearts from PVG donors and were subsequently treated with MC 1288 and CyA in various dose combinations. Administration of MC 1288 0.1 microgram/kg together with CyA 5 mg/kg produced significantly prolonged graft survival times as compared with single therapy with MC 1288 0.1 microgram/kg (p < 0.01) or CyA 5 mg/kg (p < 0.001). MC 1288 and CyA were tested also in combination with the immunomodulating drug LS-2616, which, in several laboratories, is used as a model for the evaluation of new combinations of immunosuppressive drugs. LS-2616 abolishes the immunosuppressive effect of CyA and the transplanted hearts are thus rejected at the same time as grafts of untreated recipients, i.e. on day 8. The immunosuppressive effect of MC 1288 is also counteracted by LS-2616, but the effect is not absolute and the median graft survival time is 11.0 days. Combined treatment with MC 1288 and CyA in the presence of LS-2616 resulted in a median graft survival time of 14.0 days. The results obtained in our experiments indicate at least an additive effect of MC 1288 and CyA thus suggesting that in the future MC 1288, or other vitamin D analogues, might be used to control rejection, either alone, in combination with CyA or in combination with other immunosuppressive drugs with other mechanisms of action.

Ex vivo propagation and characterization of lymphocytes from rejecting rat-kidney allografts.

Today, most clinically used methods for analysis of alloreactivity in organ transplantation are based on humoral immunity. In order to study the cellular alloresponse, a rat kidney transplantation model with culturing of graft infiltrating lymphocytes was developed. Kidney transplantations between inbred rat strains were performed with the animals initially immunosuppressed with cyclosporine. In order to initiate acute cellular rejection, immunosuppression was withdrawn after 10 days. Infiltrating lymphocytes were analysed using an in vitro culture system, allowing cells to propagate from the biopsies to culture medium. The propagated cells were counted and analysed for subtype activation markers and donor-specificity using flow cytometry and a proliferation assay. Syngeneically transplanted animals and animals given constant immunosuppression upon transplantation were used as controls. During rejection, significantly more T lymphocytes were propagating from the biopsies compared to controls. A higher percentage of the propagated T lymphocytes in the rejection group expressed activation markers [CD25 and major histocompatibility complex (MHC) class II antigen] compared to spleen- and peripheral blood T lymphocytes from the same individuals. Propagated mononuclear cells from biopsies in the rejection group were proliferating and showed donor-specific reactivity whereas mononuclear spleen cells from animals in the same group did not show this donor specificity. In conclusion, we have presented a rat kidney allotransplantation model with in vitro propagation of graft-infiltrating, activated and donor-specific T lymphocytes. This technique offers a possibility to study cellular reactivity in allotransplantation.

Synthesis and biological evaluation of technetium(III) mixed-ligand complexes with high affinity for the cerebral 5-HT(1A) receptor and the alpha1-adrenergic receptor.

Tc(III) and Re(III) complexes [M(NS(3))(CNR)] (M = Re, 99mTc, NS(3) = 2,2',2"-nitrilotris(ethanethiol), CNR = functionalized isocyanide bearing a derivative of WAY 100635) have been synthesized and characterized. Re was used as Tc surrogate for chemical characterization and in vitro receptor-binding studies. For two representatives subnanomolar affinities for the 5-HT(1A) as well as for the alpha1-adrenergic receptor were reached. Biodistribution studies in rats of the 99mTc complexes showed brain uptakes between 0.3 and 0.5% ID/organ (5 min p.i.). In vitro autoradiography of one 99mTc representative in sections of post mortem human brain indicate its accumulation in 5-HT(1A) receptor-rich brain regions. However, addition of the specific 5-HT(1A) receptor agonist 8-OH-DPAT as well as the alpha1-adrenoceptor antagonist prazosin could not substantially block this tracer accumulation. A preliminary SPET study in a monkey showed negligible brain uptake.

Bacterial translocation from defunctionalized rat small bowel.

Bacterial translocation from the intestine may cause severe infectious complications in a number of clinical situations, including the short bowel syndrome and after small bowel transplantation. The aim of the present study was to develop a simplified model for the study of bacterial translocation from a defunctionalized intestine. An ileal segment from untreated or cyclosporine-treated rats was exteriorized as a Thiry-Vella loop. After 1, 3. or 7 days, bacterial translocation and distribution of immunocompetent cells were assessed. The data obtained were compared with data from animals subjected to intestinal transplantation. Translocation to the mesenteric lymph nodes was detected in 60% of the Thiry-Vella loop animals on day 1. in 100% on day 3, and in 83% on day 7: concomitantly, the number of macrophages and T-cells in the mesenteric lymph nodes increased from day I until day 7. The degree of bacterial translocation on days 3 and 7 in animals with a Thiry-Vella loop was comparable with that observed 7 days after intestinal transplantation. Furthermore, treatment with cyclosporine A enhanced the number of translocating bacteria. In the model presented here bacterial translocation occurs from the small bowel to the mesenteric lymph nodes. The model offers possibilities to study the mechanisms and immunological phenomena associated with microbial translocation.

Hyaluronidase can be used to reduce interstitial edema in the presence of heparin.

BACKGROUND: Treatment with the hyaluronan-degrading enzyme, hyaluronidase, reduces rejection-induced interstitial edema of transplanted organs. Hyaluronidase has also been demonstrated to reduce tissue necrosis after experimentally induced myocardial infarction, but its clinical use has been limited by an observed interaction between heparin and hyaluronan. In the present work, we investigated whether it is also possible to retain the effect of the enzyme in heparinized animals. METHODS: Day 5 after heterotopic heart transplantation, recipient rats received a 2-hour intravenous infusion of hyaluronidase, either of ovine or of bovine origin. Concomitantly, the animals received intravenous heparin, either as 2 bolus doses or as a constant infusion. RESULTS: Both hyaluronidase preparations effective reduced the hyaluronan content as well as the water content of the rejecting cardiac grafts. The concomitant use of heparin did not hamper the positive results, neither when heparin was administered intermittently nor when it was given continuously. CONCLUSIONS: Our results in the transplantation model clearly demonstrate that hyaluronidase can be successfully used in heparinized individuals, provided that sufficient doses of the enzyme are given.

Chromium speciation in natural waters using serially connected supported liquid membranes.

A supported liquid membrane (SLM) method for the speciation of chromium has been developed. The method is based on selective extraction and enrichment of anionic Cr(VI) and cationic Cr(III) species in two serially connected SLM units. Methyltricaprylammonium chloride (Aliquat) and di-(2-ethylhexyl) phosphoric acid (DEHPA), respectively were used as the selective extractants in the membrane liquid. Graphite furnace atomic absorption spectrometry (GFAAS) was utilised for final determination. Optimised conditions for the DEHPA membrane were, sample solution at pH 3, acceptor solution 0.1 M HNO(3) and 10% w/w carrier in kerosene. The corresponding values for the Aliquat membrane were pH 7, 0.75 M HNO(3) and 6% w/w carrier in di-n-hexylether. This gave extraction efficiencies for Cr(III) and Cr(VI) of 90 and 40%, respectively. The method was used to measure the concentration of Cr III and Cr VI in surface water from an abandoned tannery site. Storage experiments at different pH showed that preservation at neutral pH gave almost constant values over a period of one month. At acidic pH (pH=3.0) the concentration of Cr(VI) decreased rapidly while the concentration of Cr(III) increased. The detection limit, expressed as three times the standard deviation of enriched blank samples was 0.01 microg l(-1).

Evaluation of a nurse-run asthma school.

Important aims of the study were to investigate whether an educational program (the 'Asthma School') directed by a nurse led to improved knowledge of the disease, to improved self-medication and self-management and to improved, self-rated, functional status. A total of 32 patients (6 males, 26 females, mean age 43 years) was included. The following methods were used to collect the data before and one year after the Asthma School was completed; two study-specific questionnaires for collecting demographic data and measuring different aspects of the patients' knowledge of the disease and its treatment, monthly diary cards, lung-function tests (FEV1) and the Sickness Impact Profile (SIP) questionnaire. The main results of the study were an improved knowledge of the disease and its treatment, better self-management, i.e. more frequent use of the peak expiratory flow meter (PEF-meter) and use of inhaled bronchodilators on an as-required basis, fewer patients on sick-leave and a better, self-rated, physical health status. However, in spite of these encouraging results, the lungfunction was found to be unaffected, no pronounced changes in the use of asthma drugs could be found and the patients' need for medical care remained the same.

An observation instrument for assessment of work technique in patient transfer tasks.

The aim of the study was to construct an observation instrument for description and assessment of nursing personnel's work technique in patient transfer tasks with regard to musculoskeletal health and safety, and to evaluate the validity and reliability of the instrument. The instrument consists of 24 items arranged in three phases of a transfer: the preparation phase, the starting position and the actual performance. Observations are made from video recordings. A detailed description of the individual's work technique, including actions taken to prepare the transfer, the interaction with the patient and any assistant co-worker, and the motor performance of the nurse, is provided. An attempt was made to quantify the assessments, by calculating an overall score of the work technique with regard to the level of musculoskeletal hazard and safety. The validity and reliability of the instrument were evaluated on 35 video-recorded patient transfers from hospital wards. The validity and reliability were mostly satisfactory, both when evaluating the agreements between the observations of each item (i.e. kappa values > 0.40), and when evaluating the agreements between the overall scores (i.e. intraclass correlation coefficients 0.71-0.90). Further improvements to enhance the agreements are suggested.