DOCK2 Sets the Threshold for Entry into the Virtual Memory CD8+ T Cell Compartment by Negatively Regulating Tonic TCR Triggering.

The control of cytoskeletal dynamics by dedicator of cytokinesis 2 (DOCK2), a hematopoietic cell-specific actin effector protein, has been implicated in TCR signaling and T cell migration. Biallelic mutations in Dock2 have been identified in patients with a recessive form of combined immunodeficiency with defects in T, B, and NK cell activation. Surprisingly, we show in this study that certain immune functions of CD8+ T cells are enhanced in the absence of DOCK2. Dock2-deficient mice have a pronounced expansion of their memory T cell compartment. Bone marrow chimera and adoptive transfer studies indicate that these memory T cells develop in a cell-intrinsic manner following thymic egress. Transcriptional profiling, TCR repertoire analyses, and cell surface marker expression indicate that Dock2-deficient naive CD8+ T cells directly convert into virtual memory cells without clonal effector T cell expansion. This direct conversion to memory is associated with a selective increase in TCR sensitivity to self-peptide MHC in vivo and an enhanced response to weak agonist peptides ex vivo. In contrast, the response to strong agonist peptides remains unaltered in Dock2-deficient T cells. Collectively, these findings suggest that the regulation of the actin dynamics by DOCK2 enhances the threshold for entry into the virtual memory compartment by negatively regulating tonic TCR triggering in response to weak agonists.

Autoimmunity in women: an eXamination of eXisting models.

Women comprise over 80% of the affected individuals for many autoimmune conditions. Although sex-specific differences in sex hormones are thought to contribute to the female predisposition to autoimmunity, emerging evidence also suggests an intriguing role of both physiological and dysregulated X-chromosome inactivation. Furthermore, recent studies have demonstrated that many immune genes encoded on the X chromosome are expressed biallelically, and the contribution of these sex-specific differences in immune gene dosage to autoimmunity remains to be fully explored. This review highlights recent developments in this field and discusses questions that remain unanswered.

ATM damage response and XLF repair factor are functionally redundant in joining DNA breaks.

Classical non-homologous DNA end-joining (NHEJ) is a major mammalian DNA double-strand-break (DSB) repair pathway. Deficiencies for classical NHEJ factors, such as XRCC4, abrogate lymphocyte development, owing to a strict requirement for classical NHEJ to join V(D)J recombination DSB intermediates. The XRCC4-like factor (XLF; also called NHEJ1) is mutated in certain immunodeficient human patients and has been implicated in classical NHEJ; however, XLF-deficient mice have relatively normal lymphocyte development and their lymphocytes support normal V(D)J recombination. The ataxia telangiectasia-mutated protein (ATM) detects DSBs and activates DSB responses by phosphorylating substrates including histone H2AX. However, ATM deficiency causes only modest V(D)J recombination and lymphocyte developmental defects, and H2AX deficiency does not have a measurable impact on these processes. Here we show that XLF, ATM and H2AX all have fundamental roles in processing and joining DNA ends during V(D)J recombination, but that these roles have been masked by unanticipated functional redundancies. Thus, combined deficiency of ATM and XLF nearly blocks mouse lymphocyte development due to an inability to process and join chromosomal V(D)J recombination DSB intermediates. Combined XLF and ATM deficiency also severely impairs classical NHEJ, but not alternative end-joining, during IgH class switch recombination. Redundant ATM and XLF functions in classical NHEJ are mediated by ATM kinase activity and are not required for extra-chromosomal V(D)J recombination, indicating a role for chromatin-associated ATM substrates. Correspondingly, conditional H2AX inactivation in XLF-deficient pro-B lines leads to V(D)J recombination defects associated with marked degradation of unjoined V(D)J ends, revealing that H2AX has a role in this process.

Identification of Pseudomonas aeruginosa phenazines that kill Caenorhabditis elegans.

Pathogenic microbes employ a variety of methods to overcome host defenses, including the production and dispersal of molecules that are toxic to their hosts. Pseudomonas aeruginosa, a Gram-negative bacterium, is a pathogen of a diverse variety of hosts including mammals and the nematode Caenorhabditis elegans. In this study, we identify three small molecules in the phenazine class that are produced by P. aeruginosa strain PA14 that are toxic to C. elegans. We demonstrate that 1-hydroxyphenazine, phenazine-1-carboxylic acid, and pyocyanin are capable of killing nematodes in a matter of hours. 1-hydroxyphenazine is toxic over a wide pH range, whereas the toxicities of phenazine-1-carboxylic acid and pyocyanin are pH-dependent at non-overlapping pH ranges. We found that acidification of the growth medium by PA14 activates the toxicity of phenazine-1-carboxylic acid, which is the primary toxic agent towards C. elegans in our assay. Pyocyanin is not toxic under acidic conditions and 1-hydroxyphenazine is produced at concentrations too low to kill C. elegans. These results suggest a role for phenazine-1-carboxylic acid in mammalian pathogenesis because PA14 mutants deficient in phenazine production have been shown to be defective in pathogenesis in mice. More generally, these data demonstrate how diversity within a class of metabolites could affect bacterial toxicity in different environmental niches.

A biomechanical study comparing helical blade with screw design for sliding hip fixations of unstable intertrochanteric fractures.

Dynamic hip screw (DHS) is a well-established conventional implant for treating intertrochanteric fracture. However, revision surgery sometimes still occurs due to the cutting out of implants. A helical blade instead of threaded screw (DHS blade) was designed to improve the fixation power of the osteoporotic intertrochanteric fracture. In this study, the biomechanical properties of DHS blade compared to the conventional DHS were evaluated using an unstable AO/OTA 31-A2 intertrochanteric fracture model. Fifty synthetic proximal femoral bone models with such configuration were fixed with DHS and DHS blade in five different positions: centre-centre (CC), superior-centre (SC), inferior-center (IC), centre-anterior (CA), and centre-posterior (CP). All models had undergone mechanical compression test, and the vertical and rotational displacements were recorded. The results showed that DHS blade had less vertical or rotational displacement than the conventional DHS in CC, CA, and IC positions. The greatest vertical and rotational displacements were found at CP position in both groups. Overall speaking, DHS blade was superior in resisting vertical or rotational displacement in comparison to conventional DHS, and the centre-posterior position had the poorest performance in both groups.

Applying motivational framework in medical education: a self-determination theory perspectives.

BACKGROUND: The application of self-determination theory in explaining student achievement has been well-established in various contexts. However, its application to medical education, particularly in interprofessional education (IPE) remains underexplored. Understanding how students' motivation plays a role in students' engagement and achievement is essential to optimize efforts to improve learning and instruction. OBJECTIVE: This two-stage study aims to contextualize the SDT framework to IPE through the adaptation of the Basic Psychological Need Satisfaction to IPE (Study 1) and to demonstrate how SDT can be applied in IPE by examining a model of SDT constructs (Study 2) in predicting outcomes (behavioral engagement, team effectiveness, collective dedication, goal achievement). DESIGN: In Study 1 (n=996), we adapted and validated BPNS-IPE using confirmatory factor analysis and multiple linear regression using data from 996 IPE students (Chinese Medicine, Medicine, Nursing, and Pharmacy). In Study 2 (n=271), we implemented an IPE program where we integrated SDT approaches and examined the relationship of SDT constructs with IPE outcomes using multiple linear regression. RESULTS: Our data supported the three-factor structure (autonomy, competence, and relatedness) of BPNS-IPE, meeting the required model fit. Autonomy predicted team effectiveness (F=51.290, p<.05, R2=.580); competence predicted behavioral engagement (F=55.181, p<.05, R2=.598); while relatedness predicted significantly four IPE outcomes: behavioral engagement (F=55.181, p<.01, R2=.598), team effectiveness (F=51.290, p<.01, R2=.580), collective dedication (F=49.858, p<.01, R2=.573), goal achievement (F=68.713, p<.01, R2=.649). CONCLUSIONS: The SDT motivational framework can be adapted and applied in the IPE context to understand and enhance student motivation in medical education. Potential studies with the use of the scale are provided to guide researchers.

Extrafollicular IgD-CD27-CXCR5-CD11c- DN3 B cells infiltrate inflamed tissues in autoimmune fibrosis and in severe COVID-19.

Although therapeutic B cell depletion dramatically resolves inflammation in many diseases in which antibodies appear not to play a central role, distinct extrafollicular pathogenic B cell subsets that accumulate in disease lesions have hitherto not been identified. The circulating immunoglobulin D (IgD)-CD27-CXCR5-CD11c+ DN2 B cell subset has been previously studied in some autoimmune diseases. A distinct IgD-CD27-CXCR5-CD11c- DN3 B cell subset accumulates in the blood both in IgG4-related disease, an autoimmune disease in which inflammation and fibrosis can be reversed by B cell depletion, and in severe COVID-19. These DN3 B cells prominently accumulate in the end organs of IgG4-related disease and in lung lesions in COVID-19, and double-negative B cells prominently cluster with CD4+ T cells in these lesions. Extrafollicular DN3 B cells may participate in tissue inflammation and fibrosis in autoimmune fibrotic diseases, as well as in COVID-19.

Students' interaction anxiety and social phobia in interprofessional education in Hong Kong: mapping a new research direction.

BACKGROUND: Interprofessional education (IPE) has been promoted as a breakthrough in healthcare because of the impact when professionals work as a team. However, despite its inception dating back to the 1960s, its science has taken a long time to advance. There is a need to theorize IPE to cultivate creative insights for a nuanced understanding of IPE. This study aims to propose a research agenda on social interaction by understanding the measurement scales used and guiding researchers to contribute to the discussion of social processes in IPE. METHOD: This quantitative research was undertaken in a cross-institutional IPE involving 925 healthcare students (Medicine, Nursing, Social Work, Chinese Medicine, Pharmacy, Speech Language Pathology, Clinical Psychology, Food and Nutritional Science and Physiotherapy) from two institutions in Hong Kong. Participants completed the Social Interaction Anxiety Scale (SIAS-6) and Social Phobia Scale (SPS-6). We applied a construct validation approach: within-network and between-network validation. We performed confirmatory factors analysis, t-test, analysis of variance and regression analysis. RESULTS: CFA results indicated that current data fit the a priori model providing support to within-network validity [RMSEA=.08, NFI=.959, CFI=.965, IFI=.965, TLI=.955]. The criteria for acceptable fit were met. The scales were invariant between genders, across year levels and disciplines. Results indicated that social interaction anxiety and social phobia negatively predicted behavioural engagement (F = 25.093, p<.001, R2=.065) and positively predicted behavioural disaffection (F = 22.169, p<.001, R2=.057) to IPE, suggesting between-network validity. CONCLUSIONS: Our data provided support for the validity of the scales when used among healthcare students in Hong Kong. SIAS-6 and SPS-6 have sound psychometric properties based on students' data in Hong Kong. We identified quantitative, qualitative and mixed methods research designs to guide researchers in getting involved in the discussion of students' social interactions in IPE.Key MessagesThe Social Anxiety Scale (SIAS-6) and Social Phobia Scale (SPS-6) scales have sound psychometric properties based on the large-scale healthcare students' data in IPE in Hong Kong.Social interaction anxiety and social phobia negatively predicted students' behavioural engagement with IPE and positively predicted behavioural disaffection. The scales are invariant in terms of gender, year level and discipline.Quantitative, qualitative and mixed methods studies are proposed to aid researchers to contribute in healthcare education literature using the SIAS-6 and SPS-6.

B1a and B2 cells are characterized by distinct CpG modification states at DNMT3A-maintained enhancers.

The B1 and B2 lineages of B cells contribute to protection from pathogens in distinct ways. The role of the DNA CpG methylome in specifying these two B-cell fates is still unclear. Here we profile the CpG modifications and transcriptomes of peritoneal B1a and follicular B2 cells, as well as their respective proB cell precursors in the fetal liver and adult bone marrow from wild-type and CD19-Cre Dnmt3a floxed mice lacking DNMT3A in the B lineage. We show that an underlying foundational CpG methylome is stably established during B lineage commitment and is overlaid with a DNMT3A-maintained dynamic methylome that is sculpted in distinct ways in B1a and B2 cells. This dynamic DNMT3A-maintained methylome is composed of novel enhancers that are closely linked to lineage-specific genes. While DNMT3A maintains the methylation state of these enhancers in both B1a and B2 cells, the dynamic methylome undergoes a prominent programmed demethylation event during B1a but not B2 cell development. We propose that the methylation pattern of DNMT3A-maintained enhancers is determined by the coincident recruitment of DNMT3A and TET enzymes, which regulate the developmental expression of B1a and B2 lineage-specific genes.

Induction of metabolic quiescence defines the transitional to follicular B cell switch.

Transitional B cells must actively undergo selection for self-tolerance before maturing into their resting follicular B cell successors. We found that metabolic quiescence was acquired at the follicular B cell stage in both humans and mice. In follicular B cells, the expression of genes involved in ribosome biogenesis, aerobic respiration, and mammalian target of rapamycin complex 1 (mTORC1) signaling was reduced when compared to that in transitional B cells. Functional metabolism studies, profiling of whole-cell metabolites, and analysis of cell surface proteins in human B cells suggested that this transition was also associated with increased extracellular adenosine salvage. Follicular B cells increased the abundance of the cell surface ectonucleotidase CD73, which coincided with adenosine 5'-monophosphate-activated protein kinase (AMPK) activation. Differentiation to the follicular B cell stage in vitro correlated with surface acquisition of CD73 on human transitional B cells and was augmented with the AMPK agonist, AICAR. Last, individuals with gain-of-function PIK3CD (PI3Kδ) mutations and increased pS6 activation exhibited a near absence of circulating follicular B cells. Together, our data suggest that mTORC1 attenuation may be necessary for human follicular B cell development. These data identify a distinct metabolic switch during human B cell development at the transitional to follicular stages, which is characterized by an induction of extracellular adenosine salvage, AMPK activation, and the acquisition of metabolic quiescence.

Both live and dead Enterococci activate Caenorhabditis elegans host defense via immune and stress pathways.

The innate immune response of the nematode Caenorhabditis elegans has been extensively studied and a variety of Toll-independent immune response pathways have been identified. Surprisingly little, however, is known about how pathogens activate the C. elegans immune response. Enterococcus faecalis and Enterococcus faecium are closely related enterococcal species that exhibit significantly different levels of virulence in C. elegans infection models. Previous work has shown that activation of the C. elegans immune response by Pseudomonas aeruginosa involves P. aeruginosa-mediated host damage. Through ultrastructural imaging, we report that infection with either E. faecalis or E. faecium causes the worm intestine to become distended with proliferating bacteria in the absence of extensive morphological changes and apparent physical damage. Genetic analysis, whole-genome transcriptional profiling, and multiplexed gene expression analysis demonstrate that both enterococcal species, whether live or dead, induce a rapid and similar transcriptional defense response dependent upon previously described immune signaling pathways. The host response to E. faecium shows a stricter dependence upon stress response signaling pathways than the response to E. faecalis. Unexpectedly, we find that E. faecium is a C. elegans pathogen and that an active wild-type host defense response is required to keep an E. faecium infection at bay. These results provide new insights into the mechanisms underlying the C. elegans immune response to pathogen infection.

The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo.

Distinct T follicular helper (TFH) subsets that influence specific class-switching events are assumed to exist, but the accumulation of isotype-specific TFH subsets in secondary lymphoid organs (SLOs) and tertiary lymphoid organs has not been hitherto demonstrated. IL-4-expressing TFH cells are surprisingly sparse in human SLOs. In contrast, in IgG4-related disease (IgG4-RD), a disorder characterized by polarized Ig class switching, most TFH cells in tertiary and SLOs make IL-4. Human IL-4+ TFH cells do not express GATA-3 but express nuclear BATF, and the transcriptomes of IL-4-secreting TFH cells differ from both PD1hi TFH cells that do not secrete IL-4 and IL-4-secreting non-TFH cells. Unlike IgG4-RD, IL-4+ TFH cells are rarely found in tertiary lymphoid organs in Sjögren's syndrome, a disorder in which IgG4 is not elevated. The proportion of CD4+IL-4+BATF+ T cells and CD4+IL-4+CXCR5+ T cells in IgG4-RD tissues correlates tightly with tissue IgG4 plasma cell numbers and plasma IgG4 levels in patients but not with the total plasma levels of other isotypes. These data describe a disease-related TFH subpopulation in human tertiary lymphoid organs and SLOs that is linked to IgG4 class switching.

Screw length in volar locking plate fixation for distal radial fractures.

PURPOSE: To develop a reference for the distal screw length in volar locking plate fixation for distal radial fractures in an East Asian population. METHODS: 12 pairs of forearm specimens from 11 male and one female East Asian cadavers were scanned using computed tomography. On sagittal images of the distal radius, the mean cortex-to-cortex distance of 8 quadrants was measured as a reference for the distal screw length. In addition, intra-operative 3-dimensional fluoroscopy of 10 male and 10 female patients who underwent volar locking plate fixation for distal radial fractures was used to validate the distal screw length in the cadaveric reference. 76 distal locking screws were applied in the 8 quadrants; their cortex-to-cortex distances were measured. RESULTS: The mean cortex-to-cortex distances at quadrants A, B, C, D, E, F, G, and H were 15.4 mm, 19.6 mm, 20.8 mm, 20.0 mm, 13.3 mm, 18.0 mm, 18.8 mm, and 17.4 mm, respectively. In 45% of the specimens, the 2 screws inserted at quadrants C and D were longest. Distal screws (quadrants A to D) were significantly longer than proximal screws (quadrants E to H) [p=0.02]. In intra-operative 3-dimensional fluoroscopic images, 2 of the 76 distal locking screws penetrated the dorsal cortex (one in quadrant A and one in quadrant F). The mean screw length was 88.0% of the cortex-to-cortex distance. Referenced to the cadaveric data, 88.2% of the screws could be safely inserted without penetrating the dorsal cortex, and the remaining 11.8% of screws (5 at quadrant D, one at quadrant B, and 3 at quadrant C; all in female patients) could potentially cause dorsal cortex penetration of 2 to 4 mm. In male patients, the mean screw length was 76.1% of the cortex-to-cortex distance based on the cadaveric reference. In female patients, when the screw length was 4 mm less than the cadaveric reference, the mean screw length would be 72.0% of the referenced cortex-to-cortex distance, with no dorsal cortex penetration. CONCLUSION: In female patients, the screw length should be 4 mm less than the cadaveric reference to avoid dorsal cortex penetration.

Enterococcus infection biology: lessons from invertebrate host models.

The enterococci are commensals of the gastrointestinal tract of many metazoans, from insects to humans. While they normally do not cause disease in the intestine, they can become pathogenic when they infect sites outside of the gut. Recently, the enterococci have become important nosocomial pathogens, with the majority of human enterococcal infections caused by two species, Enterococcus faecalis and Enterococcus faecium. Studies using invertebrate infection models have revealed insights into the biology of enterococcal infections, as well as general principles underlying host innate immune defense. This review highlights recent findings on Enterococcus infection biology from two invertebrate infection models, the greater wax moth Galleria mellonella and the free-living bacteriovorous nematode Caenorhabditis elegans.

The role of Candida albicans SPT20 in filamentation, biofilm formation and pathogenesis.

Candida albicans is a ubiquitous fungus, which can cause very serious and sometimes life-threatening infections in susceptible patients. We used Caenorhabditis elegans as a model host to screen a library of C. albicans mutants for decreased virulence and identified SPT20 as important for virulence. The transcription co-activator SPT20 was identified originally as a suppressor of Ty and solo δ insertion mutations, which can cause transcription defects in Saccharomyces cerevisiae. It is resistant to the toxicity caused by overexpression of GAL4-VP16. We constructed a C. albicans spt20Δ/Δ mutant and found the spt20Δ/Δ strain was significantly less virulent than the wild-type strain SC5314 in C. elegans (p < 0.0001), Galleria mellonella (p < 0.01) and mice (p < 0.001). Morphologically, spt20Δ/Δ mutant cells demonstrated a "snow-flake" shape and clustered together; prolonged culture times resulted in increased size of the cluster. The clustered morphology was associated with defects in nuclei distribution, as the nuclei were not observed in many cellular compartments. In addition, the C. albicans spt20Δ/Δ mutant resulted in defects in hyphae and biofilm formation (compared to the wild-type strain, p < 0.05), and sensitivity to cell wall and osmotic stressors, and to antifungal agents. Thus our study demonstrated a role of C. albicans SPT20 in overall morphology and distribution of nuclear material, which may cause the defects in filamentation and biofilm formation directly when this gene is deleted.

Dynamic hip screw blade fixation for intertrochanteric hip fractures.

PURPOSE: To assess the one-year outcome of a dynamic hip screw (DHS) blade in the treatment of AO/OTA 31-A1 and 31-A2 intertrochanteric hip fractures. METHODS: 35 men and 65 women aged 47 to 100 (mean, 83) years underwent fixation with a DHS blade for A1 (n=47) and A2 (n=53) intertrochanteric hip fractures after a low-energy injury. Patients were operated on within 48 hours of admission. Anteroposterior and lateral radiographs were examined for the tip-apex distance and femoral shortening. Potential complications were looked for, including implant migration, cut-out, loosening, or breakage. Functional outcome was based on the Parker mobility score. RESULTS: The 30-day and one-year mortality rates were 5% and 20%, respectively. At the one-year follow-up, 81 patients were available, and all fractures had healed without varus deformity. The mean tip-apex distance was 14.1 (range, 5.7-31.1; SD, 4.3) mm. The mean femoral shortening was 4.9 (range, 0-20.2; SD, 4.8) mm. The mean Parker score decreased to 3.8 at one-year follow-up from 5.9 before injury (p<0.001). There was one loss of fixation secondary to a non-traumatic subcapital fracture at 3 months, for which a bipolar hemiarthroplasty was performed. CONCLUSION; The DHS blade system is effective in treating AO/OTA 31-A1 and 31-A2 intertrochanteric hip fractures and results in a low complication rate.

Assessing results after distal radius fracture treatment: a comparison of objective and subjective tools.

OBJECTIVES: Functional outcomes following distal radius fractures are directly influenced by the choice of outcome assessment instruments used. Our objective was to compare scoring systems in measuring patient functional outcomes and to determine which scoring system compared most favorably with the widely used Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire. METHODS: In all, 108 patients between May 2004 and November 2006 were treated operatively following distal radius fractures. Follow-up was at 3 months, 6 months, 1 year, and 2 years postsurgery, during which anatomical and functional assessments were performed. Patient outcomes were recorded using DASH, Green and O'Brien system, Gartland and Werley system, and Sarmiento radiological scoring system. RESULTS: There was a stronger correlation between the Green and O'Brien scoring system and DASH (r = -.54) than Gartland and Werley and DASH (r = .44). The Green and O'Brien scoring system was more demanding so patients rated "excellent" or "good" had better functional outcome than those bearing the same grade in the Gartland and Werley system. Nonetheless, the Green and O'Brien score and Gartland and Werley score showed good correlation with each other (r = .66). The Sarmiento radiological score had no significant correlation with any of the other scoring systems. Significant predictors of the DASH score were function (r = .42), power grip (r = .41), pain (r = .37), and range of motion (r = .28). CONCLUSION: The Green and O'Brien scoring system correlated most strongly with the DASH score. Radiological scoring (reflecting anatomical deformity) was not significantly correlated with functional outcome. While subjective parameters "pain" and "function" are influenced by psychosocial factors and thus highly variable, it is paramount to include subjective tools in outcome assessment in future studies on wrist fractures.

Amino-terminal phosphorylation of activation-induced cytidine deaminase suppresses c-myc/IgH translocation.

Activation-induced cytidine deaminase (AID) is a mutator enzyme that initiates class switch recombination and somatic hypermutation of immunoglobulin genes (Ig) in B lymphocytes. However, AID also produces off-target DNA damage, including mutations in oncogenes and double-stranded breaks that can serve as substrates for oncogenic chromosomal translocations. AID is strictly regulated by a number of mechanisms, including phosphorylation at serine 38 and threonine 140, which increase activity. Here we show that phosphorylation can also suppress AID activity in vivo. Serine 3 is a novel phospho-acceptor which, when mutated to alanine, leads to increased class switching and c-myc/IgH translocations without affecting AID levels or catalytic activity. Conversely, increasing AID phosphorylation specifically on serine 3 by interfering with serine/threonine protein phosphatase 2A (PP2A) leads to decreased class switching. We conclude that AID activity and its oncogenic potential can be downregulated by phosphorylation of serine 3 and that this process is controlled by PP2A.

A study of constraint-induced movement therapy in subacute stroke patients in Hong Kong.

OBJECTIVE: To investigate the beneficial effect of constraint-induced movement therapy in improving the function of hemiplegic upper extremity in the early subacute stroke patients. DESIGN: A prospective, single-blinded, randomized controlled study comparing the effectiveness of constraint-induced movement therapy or control treatment at post intervention and 12 weeks follow-up. SUBJECTS: The inclusion criteria were 2-16 weeks after stroke, hemiparesis of the affected limb, minimal function of > or =20 degrees wrist extension and > or =10 degrees extension of all digits and Mini-Mental State Examination score > or =17. INTERVENTIONS: The intervention group underwent a programme of 10 days upper extremity training (4 hours per day) with the unaffected limb being restrained ina shoulder sling and the control group received an equivalent duration of conventional rehabilitation therapy. MAIN MEASURES: Functional level for hemiparetic upper extremity, Motor Activity Log, Action Research Arm Test and modified Barthel Index. RESULTS: There were 23 and 20 subjects respectively in the constraint-induced movement therapy and control groups. Significant improvements were seen at post intervention and 12 weeks after constraint-induced movement therapy in functional level for hemiparetic upper extremity (P= 0.001), and in the ;amount of use' (P= 0.001) and ;how well' (P= 0.021) subscales of the Motor Activity Log. The total Action Research Arm Test score, grasp (P= 0.004), grip (P= 0.004), pinch (P= 0.032) and gross (P= 0.006) components showed significant improvement over the control group at post intervention. The grip component (P=0.019) and the total Action Research Arm Test score (P= 0.009) were superior to the control group at 12 weeks. CONCLUSION: Significant improvement in hand function could be achieved with constraint-induced movement therapy in subacute stroke patients, which was maintained up to 12 week follow-up.