Increased frequency of proangiogenic tunica intima endothelial kinase 2 (Tie2) expressing monocytes in individuals with type 2 diabetes mellitus.

BACKGROUND: Individuals with type 2 diabetes mellitus (T2DM) have an increased risk for developing macrovascular disease (MVD) manifested by atherosclerosis. Phenotypically and functionally different monocyte subsets (classical; CD14++CD16-, non-classical; CD14+CD16++, and intermediate; CD14++CD16+) including pro-angiogenic monocytes expressing Tie2 (TEMs) can be identified. Here we investigated monocyte heterogeneity and its association with T2DM and MVD. METHODS: Individuals with (N = 51) and without (N = 56) T2DM were recruited and allocated to "non-MVD" or "with MVD" (i.e., peripheral or coronary artery disease) subgroups. Blood monocyte subsets were quantified based on CD14, CD16 and Tie2 expression levels. Plasma levels of Tie2-ligands angiopoietin-1 and angiopoietin-2 were determined using ELISA. Carotid endarterectomy samples from individuals with (N = 24) and without (N = 22) T2DM were stained for intraplaque CD68+ macrophages (inflammation) and CD34+ (angiogenesis), as plaque vulnerability markers. RESULTS: Monocyte counts were similar between individuals with T2DM and healthy controls (non-diabetic, non-MVD). Non-classical monocytes were reduced (p < 0.05) in T2DM, whereas the percentage of TEMs within the intermediate subset was increased (p < 0.05). T2DM was associated with increased angiopoietin-1 (p < 0.05) and angiopoietin-2 (p = 0.0001) levels. Angiopoietin-2 levels were higher in T2DM individuals with MVD compared with non-MVD (p < 0.01). Endarterectomized plaques showed no differences in macrophage influx and microvessel number between individuals with and without T2DM. CONCLUSIONS: Monocyte subset distribution is altered in T2DM with reduced non-classical monocytes and increased TEM percentage in the intermediate monocyte subset. Increased angiopoietin-2 levels together with increased frequency of TEMs might promote plaque vulnerability in T2DM which could however not be confirmed at tissue level in advanced atherosclerotic lesions.

CD16⁺ monocytes with smooth muscle cell characteristics are reduced in human renal chronic transplant dysfunction.

In chronic transplant dysfunction (CTD), persistent (allo)immune-mediated inflammation eventually leads to tissue remodeling including neointima formation in intragraft arteries. We previously showed that recipient-derived neointimal α-SMA(+) smooth muscle-like cells are present in human renal allografts with CTD. Human PBMC contain myeloid cells capable of differentiating into α-SMA(+) cells in vitro; the phenotype of the ancestral subset is as yet unknown. This study aimed to investigate whether monocyte subsets contain cells with smooth muscle-like cell differentiation capacity and whether CTD in renal transplant recipients is associated with a shift in these monocyte subsets. To accomplish this goal, monocyte subsets from healthy controls were sorted based on CD14 and CD16 expression to investigate gene expression levels of mesenchymal markers α-SMA and SM22α. CD14(+)/CD16(++) monocytes displayed increased α-SMA and SM22α mRNA expression compared with CD14(++)/CD16(-) monocytes, suggesting increased differentiation potential toward smooth muscle-like cells. Flow cytometry revealed that in non-CTD transplant recipients the percentage of CD14(+)/CD16(++) monocytes was reduced, with an even further reduction in patients with CTD. To determine a potential correlation between CD14(+)/CD16(++) monocytes and α-SMA(+) cell outgrowth potential in vitro, PBMC of healthy controls and transplant recipients with and without CTD were cultured under fibrotic culture conditions, and indeed a significant correlation (p=0.0002, r=0.62) was observed. Finally, double staining for α-SMA and CD16 revealed presence of α-SMA(+)CD16(+) cells in kidney explants from CTD patients, albeit at very low numbers. Our data represent evidence that, compared to CD14(++)CD16(-) monocytes, CD14(+)CD16(++) monocytes have an increased expression of smooth muscle cell-associated genes. This monocyte subpopulation is reduced in renal transplant patients with CTD, possibly due to selective migration into the allograft.

Type 2 diabetes mellitus is associated with an imbalance in circulating endothelial and smooth muscle progenitor cell numbers.

AIMS/HYPOTHESIS: Individuals with type 2 diabetes mellitus have increased rates of macrovascular disease (MVD). Endothelial progenitor cells (EPCs), circulating angiogenic cells (CACs) and smooth muscle progenitor cells (SMPCs) are suggested to play a role in the pathogenesis of MVD. The relationship between vasoregenerative EPCs or CACs and damaging SMPCs and the development of accelerated MVD in diabetes is still unknown. We tried to elucidate whether EPC, CAC and SMPC numbers and differentiation capacities in vitro differ in patients with and without diabetes or MVD. METHODS: Peripheral blood was obtained from individuals with and without diabetes and MVD (coronary or peripheral artery disease). EPC and SMPC numbers were determined with flow cytometry. Furthermore, CAC and SMPC numbers were quantified after in vitro culture. Their in vitro differentiation capacity was investigated with real-time RT-PCR and quantitative immunofluorescence. RESULTS: In diabetic patients both EPC and CAC levels were reduced (1.3-fold [p < 0.05] and 1.5-fold [p < 0.05], respectively). CAC outgrowth from diabetic patients with MVD was reduced 1.5-fold compared with diabetic patients without MVD (p < 0.05). SMPC levels were similar between diabetic patients and healthy controls. The CAC/SMPC ratio of in vitro cultured progenitor cells was reduced 2.3-fold in samples from diabetic patients (p < 0.001). The differentiation capacity of CACs and SMPCs in vitro remained similar independently of diabetes or MVD. CONCLUSIONS/INTERPRETATION: The ratio between EPCs or CACs and SMPCs is disturbed in type 2 diabetes in favour of SMPCs. This may translate into reduced vascular repair capacity, thereby promoting MVD in type 2 diabetes.

Failure by engorged stages of the lone star tick, Amblyomma americanum, to react to assembly pheromone, guanine and uric acid.

Response to tick-exposed filter paper discs (tick excreta) by fed and non-fed stages of the lone star tick Amblyomma americanum (L.) (Acari: Ixodidae) was examined from larva to adult. By contrast with engorged stages, each non-fed stage exhibited a classic arrestment response to assembly pheromone, characterized by lack of movement, retraction of legs and formation of small groups; this response was seen in approximately 75% of ticks that made contact with pheromone-treated surfaces. Less pronounced arrestment (by approximately 66% of ticks) was elicited by (0.001-0.005 M) guanine as the active pheromonal ingredient and by uric acid, a chief excretory product of birds. Lack of response to arrestment cues post-engorgement suggests that this response is kairomonal where guanine mimics uric acid as a host cue that signals immature A. americanum's preferred bird host. The functional overlap simultaneously favours tick retention in areas with an abundance of successful ticks, signalled by a heavy bout of excretion (guanine) soon after hatching and moulting. Control significance pertains to the use of these compounds as trap arrestants, but not attractants, with advantages in effectiveness against any non-fed stage.

Bone marrow-derived myofibroblasts contribute functionally to scar formation after myocardial infarction.

Myofibroblasts play a major role in scar formation during wound healing after myocardial infarction (MI). Their origin has been thought to be interstitial cardiac fibroblasts. However, the bone marrow (BM) can be a source of myofibroblasts in a number of organs after injury. We have studied the temporal, quantitative and functional role of BM-derived (BMD) myofibroblasts in myocardial scar formation. MI was induced by permanent coronary artery ligation in mice reconstituted with EGFP or pro-Col1A2 transgenic BM. In the latter, luciferase and beta-galactosidase transgene expression mirrors that of the endogenous pro-collagen 1A2 gene, which allows for functional assessment of the recruited cells. After MI, alpha-SMA-positive myofibroblasts and collagen I gradually increased in the infarct area until day 14 and remained constant afterwards. Numerous EGFP-positive BMD cells were present during the first week post-MI, and gradually decreased afterwards until day 28. Peak numbers of BMD myofibroblasts, co-expressing EGFP and alpha-SMA, were found on day 7 post-MI. An average of 21% of the BMD cells in the infarct area were myofibroblasts. These cells constituted up to 24% of all myofibroblasts present. By in vivo IVIS imaging, BMD myofibroblasts were found to be active for collagen I production and their presence was confined to the infarct area. These results show that BMD myofibroblasts participate actively in scar formation after MI.

Contact areas in the thumb carpometacarpal joint.

The thumb carpometacarpal joint is a common site of osteoarthritis. It has been hypothesized that peaks of localized stress on the dorsoradial or volar-ulnar regions, or both, of the articular surfaces of the trapezium and metacarpal lead to erosion of cartilage and may be responsible for the progression of the disease. The objective of this study was to determine the contact areas in this joint under the functional position of lateral (key) pinch and in the extremes of range of motion of the joint. These contact areas were assessed relative to the observed sites of cartilage thinning. Eight hands from cadavers of women and five from cadavers of men were tested in vitro with the thumb under a 25 N load in the lateral pinch position, and under small muscle loads (0-5 N) with the thumb in flexion, extension, abduction, adduction, and neutral positions. Contact areas of articular surfaces of the thumb carpometacarpal joint were determined for these positions using a stereophotogrammetric technique. The lateral pinch position produced contact areas predominantly on the central, volar, and volar-ulnar regions of the trapezium and the metacarpal. In three specimens, contact areas were distinctly separated between the dorsoradial and volar-ulnar regions, and in one specimen, from a man, contact occurred exclusively on the dorsoradial region of the trapezium. Using stereophotogrammetry, maps of cartilage thickness also were determined for a subset of nine specimens. The volar-ulnar, ulnar, and dorsoradial regions of the trapezium were the most common sites of thin cartilage, and these may be sites of cartilage wear.(ABSTRACT TRUNCATED AT 250 WORDS)

Excursion of the rotator cuff under the acromion. Patterns of subacromial contact.

Nine fresh-frozen, human cadaveric shoulders were elevated in the scapular plane in two different humeral rotations by applying forces along action lines of rotator cuff and deltoid muscles. Stereophotogrammetry determined possible regions of subacromial contact using a proximity criterion; radiographs measured acromiohumeral interval and position of greater tuberosity. Contact starts at the anterolateral edge of the acromion at 0 degrees of elevation; it shifts medially with arm elevation. On the humeral surface, contact shifts from proximal to distal on the supraspinatus tendon with arm elevation. When external rotation is decreased, distal and posterior shift in contact is noted. Acromial undersurface and rotator cuff tendons are in closest proximity between 60 degrees and 120 degrees of elevation; contact was consistently more pronounced for Type III acromions. Mean acromiohumeral interval was 11.1 mm at 0 degrees of elevation and decreased to 5.7 mm at 90 degrees, when greater tuberosity was closest to the acromion. Radiographs show bone-to-bone relationship; stereophotogrammetry assesses contact on soft tissues of the subacromial space. Contact centers on the supraspinatus insertion, suggesting altered excursion of the greater tuberosity may initially damage this rotator cuff region. Conditions limiting external rotation or elevation may also increase rotator cuff compression. Marked increase in contact with Type III acromions supports the role of anterior acromioplasty when clinically indicated, usually in older patients with primary impingement.

Cellular survival and proliferation in autogenous flexor tendon grafts.

In order to investigate fibroblast survival and proliferation in autogenous flexor tendon grafts, hindlimb intrasynovial and extrasynovial donor tendons were placed within the synovial sheaths of the medial and lateral forepaw digits of 21 dogs (42 tendons) and treated with controlled early passive motion. Intravital histologic evaluations with confocal microscopy and biochemical determinations of total DNA content and DNA synthesis were carried out at 10 days, 3 weeks, and 6 weeks. Intravital staining of the extrasynovial tendon grafts demonstrated variable degrees of cellular necrosis at the earliest intervals followed by cellular repopulation with fibroblasts and neovascularization from surface vessels. In contrast, intrasynovial tendon grafts were populated predominantly by viable cells at each interval, with occasional patches of cell necrosis and fibroblast ingrowth. Total DNA content and DNA synthesis values in the intrasynovial donor tendons were significantly lower than those seen in the extrasynovial tendon grafts at each interval. Extrasynovial tendons appear to act as scaffolds, undergoing extensive cellular death followed by a rapid repair response. Findings that intrasynovial tendon fibroblasts survive the tendon grafting process suggest that the nutritional supplies and metabolic requirements of intrasynovial and extrasynovial donor tendons differ largely.

The rationale for precise management of distal radius fractures.

Fractures of the distal radius continue to be one of the most common skeletal injuries treated by orthopedic or trauma surgeons. These injuries account for one sixth of all fractures seen and treated in emergency rooms. The rapid expansion of knowledge regarding the functional anatomy of the hand and wrist, the recognition by treating physicians of the ever increasing functional demands of senior citizens, and improved methodologies of achieving and maintaining anatomic restoration of these fractures have generated a renewed interest in addressing these fractures in a more precise manner.

Arthroscopic treatment of calcific tendinitis of the shoulder.

The results of arthroscopic treatment of chronic resistant calcific tendinitis of the shoulder in twenty-three patients were evaluated. Each patient had greater than 1 year of unsuccessful nonoperative management prior to arthroscopic surgery. The average age was 49 years (range 33-60) and average follow up was 26 months (range 12-47). Subacromial bursectomy was performed in all patients. Based on follow-up radiographs, thirteen patients had partial calcium removal while nine had complete removal of calcium. Results were graded as good in eleven patients (50%) with full motion and complete pain relief, satisfactory in nine (41%) patients with full motion and occasional episodes of pain, and unsatisfactory in two (9%) patients with persistent pain. Arthroscopic calcium removal and subacromial bursectomy are reasonable alternatives in the treatment of chronic calcific tendinitis resistant to conservative treatment.

A simulation model of AIDS in San Francisco: I. Model formulation and parameter estimation.

A model is formulated for the spread of the human immunodeficiency virus (HIV) and the subsequent development of acquired immunodeficiency syndrome (AIDS) in the population of homosexual men in San Francisco. The dynamic simulation model includes sexually very active and active subpopulations, migration, and a staged progression of HIV-infected persons to AIDS and death. Numerous data sources are used to estimate parameter values in the model. In a companion paper, simulations using the model and parameter estimates are found that are consistent with HIV and AIDS incidence data.

A simulation model of AIDS in San Francisco: II. Simulations, therapy, and sensitivity analysis.

The HIV and AIDS incidences each year for homosexual men in San Francisco are estimated from data. A computer simulation model for HIV transmission dynamics and progression to AIDS is used to reconstruct the HIV epidemic. Using some a priori parameter estimates, simulations are found that give good fits to the incidence data. In the stimulations the populations is divided into risk groups whose sexual activities are found to be strongly connected. There is saturation in the high-risk group, but changes in sexual behavior are more important in obtaining adequate fits. The simulation modeling yields useful parameter estimates, but the remaining uncertainty in parameter values implies that the simulation forecasts are also uncertain. Changes in HIV incidence lead to changes in AIDS incidence about 6-10 years later. Simulation models with and without zidovudine treatment both fit the incidence data; thus the effects of therapy on AIDS incidence are unclear. The fits of the simulation model are most sensitive to the yearly migration rate, the number of stages in the progression to AIDS, and the average number of new sexual partners per month; thus better estimates of these parameters would be desirable.