Precision Measurement of Vibrational Quanta in Tritium Hydride.

Saturated absorption measurements of transitions in the (2-0) band of radioactive tritium hydride are performed with the ultrasensitive noise-immune cavity-enhanced optical-heterodyne molecular spectroscopy intracavity absorption technique in the range 1460-1510 nm. The hyperfine structure of rovibrational transitions of tritium hydride, in contrast to that of hydrogen deuteride, exhibits a single isolated hyperfine component, allowing for the accurate determination of hyperfineless rovibrational transition frequencies, resulting in R(0)=203 396 426 692(22) kHz and R(1)=205 380 033 644(21) kHz. This corresponds to an accuracy 3 orders of magnitude better than previous measurements in tritiated hydrogen molecules. Observation of an isolated component in P(1) with reversed signal amplitude contradicts models for line shapes in hydrogen deuteride based on crossover resonances.

Autoantibodies against zinc transporter 8 further stratify the autoantibody-defined risk for type 1 diabetes in a general population of schoolchildren and have distinctive isoform binding patterns in different forms of autoimmune diabetes: results from the Karlsburg Type 1 Diabetes Risk Study.

AIMS: To evaluate the diagnostic relevance of autoantibodies against zinc transporter 8 (ZnT8) in schoolchildren from the general population as well as in people with autoimmune diabetes. METHODS: A total of 137 schoolchildren positive for at least one of the three major diabetes-associated autoantibodies, without diabetes heredity or preselection on HLA typing, from the Karlsburg Type 1 Diabetes Risk Study, as well as 102 people at type 1 diabetes onset, 88 people with latent autoimmune diabetes in adults and 119 people with type 2 diabetes, were analysed for different ZnT8 autoantibody variants. RESULTS: Zinc transporter 8 autoantibody positivity was found in 18% of autoantibody-positive schoolchildren, with a noticeable association with other autoantibodies associated with type 1 diabetes and disease progression. Furthermore, ZnT8 autoantibody positivity was associated with diabetes progression in schoolchildren positive for autoantibodies against insulinoma-associated antigen-2 (IA-2) and, importantly, in seven IA-2 autoantibody-negative schoolchildren. Additionally, ZnT8 autoantibodies were found in 56% of people with type 1 diabetes, predominantly directed against all three ZnT8 variants and comparable to schoolchildren with multiple autoantibodies. In contrast, ZnT8 autoantibodies were detected in 10% of people with latent autoimmune diabetes in adults, none of them with reactivity to all three isoforms. CONCLUSION: Zinc transporter 8 autoantibodies are useful markers for prediction of type 1 diabetes in a general population, further stratifying the risk of progression in autoantibody-positive children. ZnT8 autoantibodies are also important markers in adult-onset diabetes, with a completely different reaction pattern in type 1 diabetes in comparison to latent autoimmune diabetes in adults, and may therefore help to differentiate between the two forms.

Precision measurement of the fundamental vibrational frequencies of tritium-bearing hydrogen molecules: T2, DT, HT.

High-resolution coherent Raman spectroscopic measurements of all three tritium-containing molecular hydrogen isotopologues T2, DT and HT were performed to determine the ground electronic state fundamental Q-branch (v = 0 → 1, ΔJ = 0) transition frequencies at accuracies of 0.0005 cm-1. An over hundred-fold improvement in accuracy over previous experiments allows the comparison with the latest ab initio calculations in the framework of non-adiabatic perturbation theory including nonrelativisitic, relativisitic and QED contributions. Excellent agreement is found between experiment and theory, thus providing a verification of the validity of the NAPT-framework for these tritiated species. While the transition frequencies were corrected for ac-Stark shifts, the contributions of non-resonant background as well as quantum interference effects between resonant features in the nonlinear spectroscopy were quantitatively investigated, also leading to corrections to the transition frequencies. Methods of saturated CARS with the observation of Lamb dips, as well as the use of continuous-wave radiation for the Stokes frequency were explored, that might pave the way for future higher-accuracy CARS measurements.

Relativistic and QED Effects in the Fundamental Vibration of T_{2}.

The hydrogen molecule has become a test ground for quantum electrodynamical calculations in molecules. Expanding beyond studies on stable hydrogenic species to the heavier radioactive tritium-bearing molecules, we report on a measurement of the fundamental T_{2} vibrational splitting (v=0→1) for J=0-5 rotational levels. Precision frequency metrology is performed with high-resolution coherent anti-Stokes Raman spectroscopy at an experimental uncertainty of 10-12 MHz, where sub-Doppler saturation features are exploited for the strongest transition. The achieved accuracy corresponds to a 50-fold improvement over a previous measurement, and it allows for the extraction of relativistic and QED contributions to T_{2} transition energies.

Cyclooxygenase-2 contributes to the selective induction of cell death by the endocannabinoid 2-arachidonoyl glycerol in hepatic stellate cells.

The endogenous cannabinoid 2-arachidonoyl glycerol (2-AG) is an anti-fibrotic lipid mediator that induces apoptosis in hepatic stellate cells (HSCs), but not in hepatocytes. However, the exact molecular mechanisms of this selective induction of HSC death are still unresolved. Interestingly, the inducible isoform of cyclooxygenase, COX-2, can metabolize 2-AG to pro-apoptotic prostaglandin glycerol esters (PG-GEs). We analyzed the roles of COX-2 and endocannabinoid-derived PG-GEs in the differential susceptibility of primary activated HSCs and hepatocytes toward 2-AG-induced cell death. HSCs displayed significant COX-2 expression in contrast to hepatocytes. Similar to 2-AG, treatment of HSCs with PGD2-GE dose-dependently induced cell death independently from cannabinoid receptors that was accompanied by PARP- and caspase 3-cleavage. In contrast to 2-AG, PGD2-GE failed to induce significant ROS formation in HSCs, and depletion of membrane cholesterol did not rescue HSCs from PGD2-GE-induced apoptosis. These findings indicate differential engagement of initial intracellular signaling pathways by 2-AG and its COX-2-derived metabolite PGD2-GE, but similar final cell death pathways. Other PG-GEs, such as PGE2-or PGF2α-GE did not induce apoptosis in HSCs. Primary rat hepatocytes were mainly resistant against 2-AG- and PGD2-GE-induced apoptosis. HSCs, but not hepatocytes were able to metabolize 2-AG to PGD2-GE. As a proof of principle, HSCs from COX-2(-/-) mice lacked PDG2-GE production after 2-AG treatment. Accordingly, COX-2(-/-) HSCs were resistant against 2-AG-induced apoptosis. In conclusion, the divergent expression of COX-2 in HSCs and hepatocytes contributes to the different susceptibility of these cell types towards 2-AG-induced cell death due to the generation of pro-apoptotic PGD2-GE by COX-2 in HSCs. Modulation of COX-2-driven metabolization of 2-AG may provide a novel physiological concept allowing the specific targeting of HSCs in liver fibrosis.

Autoantibody-defined risk for Type 1 diabetes mellitus in a general population of schoolchildren: results of the Karlsburg Type 1 Diabetes Risk Study after 18 years.

AIMS: To investigate the occurrence of diabetes-associated autoantibodies and cumulative Type 1 diabetes risk over 18 years in a general population of schoolchildren. METHODS: In the Karlsburg Type 1 Diabetes Risk Study, 11 986 schoolchildren from north-eastern Germany without a family history of diabetes were screened for glutamic acid decarboxylase antibodies, insulinoma-associated antigen-2 antibodies and insulin autoantibodies by radioligand binding assay. Those children found to be autoantibody-positive were invited to follow-up examinations and HLA-DQB1 genotyping, and were followed for progression to Type 1 diabetes. RESULTS: At first follow-up, 119 children had single and 36 children had multiple autoantibodies. Of the multiple autoantibody-positive children, 33 had at least one diabetes-associated HLA-DQB1 allele (*02 and/or *0302). A total of 26 children progressed to Type 1 diabetes, of whom 22 had multiple autoantibodies. The male-to-female ratio of those who progressed to Type 1 diabetes was 1.6. The positive predictive value of multiple autoantibodies was 61.1% compared with only 23.7% for diabetes-associated HLA-DQB1 genotypes among all those who were autoantibody-positive. The cumulative risk was 59.7% after 10 years and 75.1% after 18 years for children with multiple autoantibodies compared with 1.2 and 22.6%, respectively, for children with single autoantibodies (P<0.001). Among the three examined autoantibodies, insulinoma-associated antigen-2 antibodies conferred the highest risk. CONCLUSIONS: The diabetes risk in schoolchildren with multiple autoantibodies was similar to the risk reported in other studies for genetically preselected probands; thus, a combined autoantibody-based screening could effectively identify at-risk individuals from the general population for future intervention trials.

Quantitative immunohistochemical examination of the local cellular reactions following implantation of biomaterials.

Examining the biocompatibility of implant materials includes the in vivo investigation of the local tissue response following implantation in experimental animals. By contrast to qualitative and semi-quantitative approaches often used in this field, a quantitative technique would facilitate a more accurate determination and better comparability of different studies. Therefore, this study aimed at evaluating the applicability of the free image analysis software ImageJ for fast, easy and reproducible quantification of the tissue response following implantation of titanium samples in rats with subsequent immunohistochemical examination of peri-implant tissue samples for monocytes and macrophages (ED1) and MHC class II positive antigen presenting cells (OX6). The quantification of positively stained cells in the vicinity of the implant pockets was based on a grid-supported manual count carried out using two ImageJ plugins (CellCounter, Grid) and resulted in a mean coefficient of variation of 13.8% (ED1) and 19.6% (OX6) between different investigators and 10.0% (ED1) and 13.8% (OX6) for repeated counting by the same investigator. In conclusion, ImageJ was found to be suitable for morphometric evaluation of the tissue response following implantation, particularly the analysis of discrete cellular events at the tissue-biomaterial interface. The procedure which was used is described in detail, and its advantages and disadvantages are discussed.

Diabetes Antibody Standardization Program: evaluation of assays for insulin autoantibodies.

AIMS/HYPOTHESIS: Insulin autoantibodies (IAA) are important in type 1 diabetes risk assessment. However, their determination varies more between laboratories than other diabetes autoantibodies. The Diabetes Antibody Standardization Program (DASP) aims to improve and standardise measurement of autoantibodies associated with type 1 diabetes. We report the results of measurement of IAA from DASP workshops in 2002, 2003 and 2005. METHODS: Up to 32 laboratories in 14 countries participated in each workshop. Aliquots of coded sera from 50 patients with newly diagnosed type 1 diabetes and 100 blood donor controls were circulated to participating laboratories. Reported results were analysed using receiver operator characteristic (ROC) curves. We compared concordance of antibody levels by ranking, IAA and insulin antibody (IA) indices and units derived from an IA standard curve. RESULTS: In all three workshops IAA assay performance had improved compared with DASP 2000. The median area under the ROC curve was 0.73 in DASP 2002, 0.78 in 2003 and 0.80 in 2005 (p = 0.0012), and median laboratory-assigned sensitivity was 26% in 2002, 36% in 2003 and 45% in 2005 (p < 0.0001). There was, however, marked variation between assays. The range of AUC was 0.36-0.91 and that of laboratory-assigned sensitivity was 22-57%. Concordance of ranking of patient serum samples was related to AUC (p < 0.001). Using an index related to common IAA and IA-positive or -negative control sera improved the concordance between assays (p < 0.0001). CONCLUSIONS/INTERPRETATION: The overall performance of IAA assays has improved but there is still wide variation between laboratories. Concordance between assays would be improved by the use of a common reference reagent.

Coherent transport of atomic quantum states in a scalable shift register.

We demonstrate the coherent transport of 2D arrays of small ensembles of neutral atoms in a shift register architecture based on 2D arrays of microlenses. We show the scalability of the transport process by presenting the repeated hand over of atoms from site to site. We prove the conservation of coherence during transport, reloading, and a full shift register cycle. This shows that the fundamental shift sequence can be cascaded and thus scaled to complex and versatile 2D architectures for atom-based quantum information processing, quantum simulation, and the investigation of quantum degenerate gases.

Digital laser frequency and intensity stabilization based on the STEMlab platform (originally Red Pitaya).

We report on the development, implementation, and characterization of digital controllers for laser frequency stabilization as well as intensity stabilization and control. Our design is based on the STEMlab (originally Red Pitaya) platform. The presented analog hardware interfaces provide all necessary functionalities for the designated applications and can be integrated in standard 19-in. rack mount units. Printed circuit board layouts are made available as an open-source project (T. Preuschoff et al., https://github.com/TU-Darmstadt-APQ/RedPitaya-Lockbox, 2020 and T. Preuschoff et al., https://github.com/TU-Darmstadt-APQ/RedPitaya-IntStab, 2020). A detailed characterization shows that the bandwidth (1.25 MHz) and the noise performance of the controllers are limited by the STEMlab system and not affected by the supplementary hardware. Frequency stabilization of a diode laser system resulting in a linewidth of 52(1) kHz (FWHM) is demonstrated. Intensity control to the 1 × 10-3 level with sub-microsecond rise and fall times based on an acousto-optic modulator as actuator is achieved.

The bovine protamine 2 gene: evidence for alternative splicing.

Protamine 2 (PRM2) is a low molecular weight arginine-rich protein which is present in haploid spermatogenic cells of human and mouse. Although the bull PRM2 gene is translated and transcribed at low levels, the protein could not be detected. The gene was isolated from a cosmid library and was found to consist of two exons (298 and 50 bp, respectively) interrupted by an intron of 142 bp. As compared to the PRM2 genes of man, mouse and rat the bovine gene lacks a highly conserved sequence coding for the amino acids RLHRIH. Furthermore, primer extension experiments on bull PRM2 mRNA and sequencing of junction fragments revealed alternative splicing of mRNA resulting in two putative isoforms of the protein. The most abundant transcript is spliced at the conserved splice donor site found in exon 1 at position 236 giving rise to an in-frame deletion of 63 bp as compared to the cDNA sequence (Maier et al. (1990) Nucleic Acids Res. 18, 1249-1254). The less abundant longer mRNA was not detectable by radioactive primer extension. The corresponding cDNA was obtained by performing PCR with reverse transcribed bull testis RNA or with a spermatid specific cDNA library. Alternative splicing should result in an addition of 21 nonpolar amino acids in the derived polypeptide and an altered protein conformation and function.

Enterovirus RNA sequences in sera of schoolchildren in the general population and their association with type 1-diabetes-associated autoantibodies.

Type 1 diabetes (T1D) is an autoimmune disease linked with genetic factors as well as with environmental triggers, such as virus infections, but the aetiology is still unclear. The authors analysed serum from autoantibody-positive (n=50) and autoantibody-negative (n=50) schoolchildren as well as children newly diagnosed with T1D (n=47; time from diagnosis, median 5 days, interquartile range 1-12 days) for the presence and frequency of enterovirus (EV) and adenovirus sequences. The autoantibody-positive and -negative groups were part of the Karlsburg Type 1 Diabetes Risk Study of a Normal Schoolchild Population, which represents a general population without T1D first-degree relatives. There was no significant seasonality of sampling in any of the three groups investigated. EV RNA sequences were detected in 10 of 50 (20%) autoantibody-positive children and in 17 of 47 (36%) children newly diagnosed with T1D, but only in two of 50 (4%) of the age- and sex-matched controls (P<0.05, P<0.001). Characterization of the EV amplicons by direct sequencing revealed high homology with coxsackievirus B group. For adenovirus we found no data to support an association with T1D. The data support the hypothesis that different enteroviruses may be aetiologically important as a trigger and/or accelerating factor in the process of T1D development.

Quantitative evaluation of a monoclonal antibody and its fragment as potential markers for pancreatic beta cell mass.

Antibodies, due to their high specificities and retention, represent potential beta cell imaging agents, however their slow clearance from the blood may preclude their use. Antibody fragments (Fabs) have much higher clearance and if they can be made with similar binding characteristics, would be more efficacious agents. An existing beta cell specific antibody (K14D10) and its Fab were evaluated with a previously developed screening assay. The Fab and the intact immunoglobulin (IgG) had similar affinities (6 - 20 nM), binding sites (300 000 - 700 000 sites/cell), and binding kinetics (t (1/2) = 8 - 18 minutes) for beta cells. However, the cellular specificity was far below the estimated requisite values needed to overcome the very low beta cell mass in the pancreas. The Fab cleared the blood twice as fast as the IgG, but did not preferentially accumulate into pancreas. Thus, generation of Fabs from IgGs with high beta cell binding and blood clearance appears feasible, but in order for molecules to be useful for tracking beta cell mass, antibodies of greater cellular specificity will have to be used.

Antibody response to collagen after functional implantation of different polyester vascular prostheses in pigs.

Besides inflammation, specific immune responses are seen also after implantation of biomaterials. The aim was to investigate the humoral response to bovine collagen type I following implantation of various polyester (Dacron) prostheses into pigs. In 24 randomized pigs, the infrarenal aorta was replaced with a segment of collagen-impregnated, woven polyester prosthesis of low, medium, or high porosity. IgG antibodies were detected by immunoassay using native and denatured collagen type I as a target for blood samples taken on day 1 (implantation), 10, 17, 24, 62, and 116. As generally observed, antibodies to native and denatured collagen are of low titer and were significantly correlated with enhanced binding to the denatured form (p < 0.001). The highest overall antibody prevalence to native and denatured collagen was obtained on day 116 with 68% and on day 62 with 59%, respectively. Prostheses with high porosity induced an early immune response on day 10; those with low and medium porosity induced the highest antibody levels later after 2 months. Collagen antibodies neither correlated with serum IgG contents nor with antibodies to the prosthesis polyester matrix. Thus, humoral immune response against implant components may provide a further parameter in describing biocompatibility but also a potential marker that may facilitate monitoring of individual perigraft reaction.

Influence of target cell preparation on binding of monoclonal islet cell reactive antibodies (mc-ICRA) in cellular enzyme-linked immunosorbent assay (CELISA).

A rapid, effective and sensitive CELISA for the detection of monoclonal islet cell reactive antibodies (mc-ICRA) using the insulin-producing rat insulinoma cell line (RIN) is described. RIN cells are a suitable target for this monoclonal antibody assay as shown by a comparative study with normal rat islet cells. We tested the influence of the target cell preparation and obtained the best sensitivity and reliability with the CELISA using desiccated cells or desiccated cell homogenate with a cell number of 5 x 10(4) cells per well rather than an adsorbed cell homogenate. Furthermore, ethanol fixation of RIN cells resulted in a loss of antigenicity as shown particularly by the detection of islet cell surface antibodies. We also compared the binding of mc-ICRA in RIN-CELISA with data obtained by indirect immunofluorescence using viable RIN cells as targets. By permeabilization of the cell membrane by desiccation or sonication, more antibodies are detected in CELISA (surface and cytoplasmic antibodies), whereas in immunofluorescence on viable RIN cells, only surface reactive antibodies are detected.

Metabolism of phenol and hydroquinone to reactive products by macrophage peroxidase or purified prostaglandin H synthase.

Macrophages, an important cell-type of the bone marrow stroma, are possible targets of benzene toxicity because they contain relatively large amounts of prostaglandin H synthase (PHS), which is capable of metabolizing phenolic compounds to reactive species. PHS also catalyzes the production of prostaglandins, negative regulators of myelopoiesis. Studies indicate that the phenolic metabolites of benzene are oxidized in bone marrow to reactive products via peroxidases. With respect to macrophages, PHS peroxidase is implicated, as in vivo benzene-induced myelotoxicity is prevented by low doses of nonsteroidal anti-inflammatory agents, drugs that inhibit PHS. Incubations of either 14C-phenol or 14C-hydroquinone with a lysate of macrophages collected from mouse peritoneum (greater than 95% macrophages), resulted in an irreversible binding to protein that was dependent upon H2O2, incubation time, and concentration of radiolabel. Production of protein-bound metabolites from phenol or hydroquinone was inhibited by the peroxidase inhibitor aminotriazole. Protein binding from 14C-phenol also was inhibited by 8 microM hydroquinone, whereas binding from 14C-hydroquinone was stimulated by 5 mM phenol. The nucleophile cysteine inhibited protein binding of both phenol and hydroquinone and increased the formation of radiolabeled water-soluble metabolites. Similar to the macrophage lysate, purified PHS also catalyzed the conversion of phenol to metabolites that bound to protein and DNA; this activation was both H2O2- and arachidonic acid-dependent. These results indicate a role for macrophage peroxidase, possibly PHS peroxidase, in the conversion of phenol and hydroquinone to reactive metabolites and suggest that the macrophage should be considered when assessing the hematopoietic toxicity of benzene.

Prevention of benzene-induced myelotoxicity by nonsteroidal anti-inflammatory drugs.

Benzene affects hematopoietic progenitor cells leading to bone marrow depression and genotoxic effects such as micronucleus formation. Progenitor cell proliferation and differentiation are inhibited by prostaglandins produced by macrophages. Administration of benzene to DBA/2 or C57BL/6 mice caused a dose-dependent bone marrow depression and a significant increase in marrow prostaglandin E level and both were prevented by the coadministration of indomethacin and other inhibitors of the cyclooxygenase component of prostaglandin H synthase. Levels of benzene that decreased bone marrow cellularity also caused genotoxic effects measured as increased micronucleated polychromatic erythrocytes in peripheral blood, which was also prevented by the coadministration of indomethacin. These results suggest a possible role for prostaglandin synthase in benzene myelotoxicity; a mechanism by which this might occur is presented.

A monoclonal antibody-based characterization of autoantibodies against glutamic acid decarboxylase in adults with latent autoimmune diabetes.

Autoantibodies to glutamic acid decarboxylase (GAD) are an important marker of the autoimmune-mediated beta-cell destruction in insulin-dependent (Type I) diabetes. However, these autoantibodies are also found in patients with Stiff-man syndrome (SMS) without onset of diabetes and some diabetic patients who initially present as non-insulin dependent (Type II) diabetes later becoming insulin-dependent, called as latent autoimmune diabetes in adults (LADA). To study the immune response to GAD in these LADA patients a competitive radiobinding assay based on murine monoclonal antibodies recognizing three different GAD regions was performed. The monoclonal antibodies against GAD recognize two different linear epitopes localized at the N- (amino acids 4-17) and C-terminus (amino acids 572-585) and one conformation-dependent epitope region (amino acids 221-442 IDDM-E1) known to be immunodominant for diabetes-associated autoantibodies. All LADA sera (20/20) reduced substantially the 125I-GAD binding of the monoclonal antibodies reactive with the conformation-dependent epitope region IDDM-E1 and only 20% of these sera additionally diminished the 125I-GAD65 binding by those monoclonals reactive with the both linear epitopes. The SMS sera completely abolished the GAD binding of all three monoclonals, reflecting a broader repertoire including an immune response against the IDDM-E1, a conformation-dependent GAD65 epitope region, also revealed if the SMS sera are diluted to equivalent antibody concentrations. In summary, our results show that diabetes-associated GAD autoantibodies even in adult patients with a late autoimmune process preferentially recognize a conformation-dependent middle GAD65 region. An immune response to all three GAD epitope regions is seldom in these LADA patients and only detectable in association with high antibody titres.

Autoantibodies against GAD65 rather than GAD67 precede the onset of type 1 diabetes.

The enzyme glutamate decarboxylase (GAD) is considered one of the major Beta cell antigens in Type 1 diabetes mellitus. The GAD autoantibody (GAD-AAb) prevalence in newly diagnosed Type 1 diabetic patients has been described up to 80%, depending on the detection method used. The aim of this study was to evaluate a simple, specific, and sensitive radioimmunoassay (RIA) method for detection of AAb against both isoforms of the enzyme, GAD65 and GAD67, in a cross-sectional study using sera from newly diagnosed Type 1 diabetic patients and in a longitudinal study using sera from prediabetic patients and individuals at risk of developing the disease. The 125I-labelled full-length human recombinant proteins of GAD65 and GAD67 expressed in SF9 cells were used as the antigen source. The prevalence of GAD65-AAb in newly diagnosed Type 1 diabetic patients was found to be 73% (112/153), in contrast to 19% (14/72) of GAD67-AAb. Only one patient produced AAb restricted to GAD67. Furthermore, GAD65-AAb could also be detected in 73% (11/15) of prediabetic patients (up to 122 months before clinical manifestation of the disease), whereas only 27% (4/15) of them were positive for GAD67-AAb. In the group at risk of developing Type 1 diabetes, these prevalences were 77% (10/13) and 46% (6/13), respectively. In all GAD67-AAb-positive patients investigated in the longitudinal study, AAb to GAD65 were detectable. In 47% of patients positive for both GAD65-AAb and ICA, the GAD65-AAb appeared by up to 46 months before the occurrence of ICA was detected. The data illustrated that GAD65 is the main immunogenic isoform of the enzyme in the preclinical and clinical stages. The RIA detecting AAb against this isoform may facilitate the screening for individuals at risk of developing the disease.

Long-term outcome of pulmonary rehabilitation in patients with COPD.

BACKGROUND: This study investigates the long-term benefits of pulmonary rehabilitation in terms of health-related quality of life (HRQL). Such information is of particular importance in developing strategies for aftercare at home which aim to maintain the initial improvements seen after rehabilitation. METHODS: Criteria for inclusion were diagnosis of COPD, age 40 to 80 years, and completion of an inpatient pulmonary rehabilitation program. HRQL was assessed by the St. George Respiratory Questionnaire, and the component "well-being" from the Medical Psychological Questionnaire for Lung Diseases. Patient characteristics included lung function parameters such as FEV1, the diffusion capacity for carbon monoxide and maximal inspiratory mouth pressure, age, socio-economic variables, and exercise tolerance evaluated by a 12-min walking test. To define patients in whom long-term benefits were sustained 9 months postdischarge, cases were clustered using hierarchical cluster analysis, based on the HRQL scores at discharge. RESULTS: Complete data sets were obtained from 77 patients. Two groups of cases were clustered. Patient characteristics were essentially the same in both groups. HRQL differed significantly between groups on admission, at discharge, and at follow-up. Within-group analysis revealed that patients in group 1 (n=44) had "moderate" scores on HRQL on admission, a significant improvement between admission and discharge, followed by a significant deterioration of HRQL at follow-up. Group 2 (n=33) had "severely" impaired HRQL on admission, little improvement after rehabilitation, and remained in fairly stable condition 9 months postdischarge. CONCLUSIONS: Results suggest that patients with COPD require a differentiated aftercare program of postdischarge pulmonary rehabilitation.