Early vascularization is a prerequisite for successful bone healing and endothelial progenitor cells (EPC), seeded on appropriate biomaterials, can improve vascularization. The type of biomaterial influences EPC function with bioglass evoking a vascularizing response. In this study the influence of a composite biomaterial based on polylactic acid (PLA) and either 20 or 40% bioglass, BG20 and BG40, respectively, on the differentiation and survival of EPCs in vitro was investigated. Subsequently, the effect of the composite material on early vascularization in a rat calvarial critical size defect model with or without EPCs was evaluated. Human EPCs were cultured with β-TCP, PLA, BG20 or BG40, and seeding efficacy, cell viability, cell morphology and apoptosis were analysed in vitro. BG40 released the most calcium, and improved endothelial differentiation and vitality best. This effect was mimicked by adding an equivalent amount of calcium to the medium and was diminished in the presence of the calcium chelator, EGTA. To analyze the effect of BG40 and EPCs in vivo, a 6-mm diameter critical size calvarial defect was created in rats (n = 12). Controls (n = 6) received BG40 and the treatment group (n = 6) received BG40 seeded with 5×105 rat EPCs. Vascularization after 1 week was significantly improved when EPCs were seeded onto BG40, compared to implanting BG40 alone. This indicates that Ca2+ release improves EPC differentiation and is useful for enhanced early vascularization in critical size bone defects.
Treatment of congenital adrenal hyperplasia
Leprosy reversal reactions type 1 (T1R) are acute immune episodes that affect a subset of leprosy patients and remain a major cause of nerve damage. Little is known about the relative importance of innate versus environmental factors in the pathogenesis of T1R. In a retrospective design, we evaluated innate differences in response to Mycobacterium leprae between healthy individuals and former leprosy patients affected or free of T1R by analyzing the transcriptome response of whole blood to M. leprae sonicate. Validation of results was conducted in a subsequent prospective study. We observed the differential expression of 581 genes upon exposure of whole blood to M. leprae sonicate in the retrospective study. We defined a 44 T1R gene set signature of differentially regulated genes. The majority of the T1R set genes were represented by three functional groups: i) pro-inflammatory regulators; ii) arachidonic acid metabolism mediators; and iii) regulators of anti-inflammation. The validity of the T1R gene set signature was replicated in the prospective arm of the study. The T1R genetic signature encompasses genes encoding pro- and anti-inflammatory mediators of innate immunity. This suggests an innate defect in the regulation of the inflammatory response to M. leprae antigens. The identified T1R gene set represents a critical first step towards a genetic profile of leprosy patients who are at increased risk of T1R and concomitant nerve damage.
Leprosy type 1 reversal reactions (T1R) are an important cause of nerve damage in leprosy patients and accurate prediction of patients at increased risk of T1R is a major challenge of current leprosy control. The incidence of T1R differs widely from 6% to 67% of leprosy patients in different leprosy endemic settings. Whether or not this reflects the impact of unknown environmental triggers or differences in the genetic background across ethnicities is not known. We performed a comparative transcriptome analysis between leprosy patients affected and free of T1R in response to M. leprae antigens. As the discovery sample we enrolled cured leprosy patients who had been diagnosed with T1R at the time of leprosy diagnosis and leprosy patients who had never undergone T1R (retrospective arm). Whole genome transcriptome analysis after stimulation of blood with M. leprae antigen resulted in the definition of a T1R signature gene set. We validated the T1R gene set in RNA samples obtained from T1R-free patients at the time of leprosy diagnosis and followed for 3 years for development of T1R (prospective arm). These results confirm the role of innate factors in T1R and are a first step towards a predictive genetic T1R signature.
To evaluate the clinical and functional results of a technical procedure used in the surgical treatment of congenital constriction ring (CCR) in children.
Materials and methods
This was a retrospective study undertaken to evaluate the results of surgical techniques performed from January 1995 to December 2005 on 95 patients with 134 congenital constriction bands. Due to the drop-out of nine patients during follow-up, data on 86 patients (121 congenital constriction rings; average age at surgery 1 year 2 months) were analyzed. The extent of the constrictions was classified by according to the Patterson criteria. All patients were treated by two-stage sine plasty combined with removal of the fibrous groove and fasciotomy, with one-half of the ring removed during the first stage and the other half removed 1 week later during the second state. The surgical outcomes were assess according to the Moses criteria.
Three types of CCR (Patterson criteria) were identified among the 86 patients (121 constriction rings): types I (5 patients, 4.1 %), II (107, 88.5 %), III (9, 7.4 %). Of the 121 constriction rings, good results were attained in 73.6 % and fair results in 26.4 %. Sensory deficits were seen in six patients immediately after the surgery but all six had improved to a normal condition at the final follow-up examination. There were no skin necrosis or wound healing problems.
The combined sine plasty/removal of fibrous groove and fasciotomy method reported here is a simple and safe surgical technique for treating CCR in children.
Congenital constriction band syndrome; Dermofat flap; Amniotic bands; Direct closure; Z-plasty
A public health intervention program with active involvement of local related stakeholders was piloted in the Bien Hoa dioxin hotspot (2007–2009), and then expanded to the Da Nang dioxin hotspot in Vietnam (2009–2011). It aimed to reduce the risk of dioxin exposure of local residents through foods. This article presents the results of the intervention in Da Nang.
To assess the results of this intervention program, pre- and post-intervention knowledge, attitude, and practice (KAP) surveys were implemented in 400 households, randomly selected from four wards surrounding the Da Nang Airbase in 2009 and 2011, respectively.
After the intervention, the knowledge on the existence of dioxin in food, dioxin exposure pathways, potential high-risk foods, and preventive measures significantly increased (P<0.05). Ninety-eight percent were willing to follow advice on preventing dioxin exposure. Practices to reduce the risk of dioxin exposure also significantly improved (P<0.05). After intervention, 60.4% of households undertook exposure preventive measures, significantly higher than that of the pre-intervention survey (39.6%; χ2=40.15, P<0.001). High-risk foods had quite low rates of daily consumption (from 0 to 2.5%) and were significantly reduced (P<0.05).
This is seen as an effective intervention strategy toward reducing the risk of human exposure to dioxin at dioxin hotspots. While greater efforts are needed for remediating dioxin-polluted areas inside airbases, there is also evidence to suggest that, during the past four decades, pollution has expanded to the surrounding areas. For this reason, this model should be quickly expanded to the remaining dioxin hotspots in Vietnam to further reduce the exposure risks in other areas.
dioxin hotspots; intervention program; dioxin exposure through foods; risk communication; dioxin risk reduction; Vietnam
Vascular cartilaginous metaplasia and calcification are common in patients with atherosclerosis. However, sources of cells contributing to the development of this complication are currently unknown. In this study, we ascertained the origin of cells that give rise to cartilaginous and bony elements in atherosclerotic vessels.
Methods and results
We utilized genetic fate mapping strategies to trace cells of smooth muscle (SM) origin via SM22α-Cre recombinase and Rosa26-LacZ Cre reporter alleles. In animals expressing both transgenes, co-existence within a single cell of β-galactosidase [marking cells originally derived from SM cells (SMCs)] with osteochondrogenic (Runx2/Cbfa1) or chondrocytic (Sox9, type II collagen) markers, along with simultaneous loss of SM lineage proteins, provides a strong evidence supporting reprogramming of SMCs towards osteochondrogenic or chondrocytic differentiation. Using this technique, we found that vascular SMCs accounted for ∼80% of Runx2/Cbfa1-positive cells and almost all of type II collagen-positive cells (∼98%) in atherosclerotic vessels of LDLr−/− and ApoE−/− mice. We also assessed contribution from bone marrow (BM)-derived cells via analysing vessels dissected from chimerical ApoE−/− mice transplanted with green fluorescence protein-expressing BM. Marrow-derived cells were found to account for ∼20% of Runx2/Cbfa1-positive cells in calcified atherosclerotic vessels of ApoE−/− mice.
Our results are the first to definitively identify cell sources attributable to atherosclerotic intimal calcification. SMCs were found to be a major contributor that reprogrammed its lineage towards osteochondrogenesis. Marrow-derived cells from the circulation also contributed significantly to the early osteochondrogenic differentiation in atherosclerotic vessels.
Atherosclerosis; Circulating progenitors; Osteochondrogenic differentiation; Smooth muscle cells; Vascular calcification
There is a critical public health problem in the United States today, the problem of childhood psychiatric disorders in youngsters with physical illnesses. Currently there is a pressing need for well-trained pediatric psychosomatic medicine practitioners as well as advanced training in the field. Yet, this training does not currently exist. This article will present the innovative Montefiore Medical Center/Albert Einstein College of Medicine (MMC/AECOM) program as a model for a training curriculum, clinical training experience, and clinical research training setting in this important and rapidly expanding area of need in pediatric mental health.
Leprosy is a persistent infectious disease caused by Mycobacterium leprae that still affects over 200,000 new patients annually. The host genetic background is an important risk factor for leprosy susceptibility and the PARK2 gene is a replicated leprosy susceptibility candidate gene. The protein product of PARK2, Parkin, is an E3 ubiquitin ligase that is involved in the development of various forms of Parkinsonism. The human macrophage is both a natural host cell of M. leprae as well as a primary mediator of natural immune defenses, in part by secreting important pro-inflammatory cytokines and chemokines. Here, we report that down-regulation of Parkin in THP-1 macrophages, human monocyte-derived macrophages and human Schwann cells resulted in a consistent and specific decrease in interleukin-6 (IL-6) and monocyte chemoattractant protein 1 (MCP-1/CCL2) production in response to mycobacteria or LPS. Interestingly, production of IL-6 at 6 hours by THP-1 cells stimulated with live M. leprae and M. bovis BCG was dependent on pretreatment with 1,25-dihydroxyvitamin D3 (VD). Parkin knockdown in VD-treated cells blocked IL-6 induction by mycobacteria. However, IκB-α phosphorylation and levels of IκB-ξ, a nuclear protein required for IL-6 expression, were not affected by Parkin silencing. Phosphorylation of MAPK ERK1/2 and p38 was unaffected by Parkin silencing while JNK activation was promoted but did not explain the altered cytokine production. In a final set of experiments we found that genetic risk factors of leprosy located in the PARK2 promoter region were significantly correlated with M. leprae sonicate triggered CCL2 and IL6 transcript levels in whole blood assays. These results associated genetically controlled changes in the production of MCP-1/CCL2 and IL-6 with known leprosy susceptibility factors.
Leprosy is an infectious disease with a strong host genetic component. The identification of host genetic lesions predisposing to disease is a powerful approach for mapping key junctions in the host pathogen interplay. Genetic variants located in the promoter region of the PARK2 gene are replicated leprosy susceptibility factors. To better understand a possible contribution of PARK2 to host effector mechanisms in leprosy patients, we developed a cellular model to test the contribution of the PARK2 encoded parkin protein to host responses to mycobacterial antigens. We observed that parkin was a mediator of IL-6 production in response to mycobacterial antigen in both THP-1 macrophages and human Schwann cells while human monocyte-derived macrophages needed to be pre-activated with VitD to show the same impact. Parkin also impacted on the constitutive production of MCP-1. The regulatory activity of parkin on cytokine production was found to be independent of the canonical TLR-NFκB signalling pathway. We also tested association of IL6 and CCL2 gene expression levels in whole blood assays with PARK2 polymorphisms. For both cytokines, we found significant associations with those PARK2 variants that were established leprosy susceptibility factors. Hence, our results show that genetic PARK2 variants that are correlated with leprosy susceptibility are also correlated with production of these cytokines following stimulation with M. leprae sonicate.
To study the individual functions of hyaluronan interacting proteins in prostate cancer (PCa) motility through connective tissues, we developed a novel three-dimensional (3D) hyaluronic acid (HA) hydrogel assay that provides a flexible, quantifiable, and physiologically relevant alternative to current methods. Invasion in this system reflects the prevalence of HA in connective tissues and its role in the promotion of cancer cell motility and tissue invasion, making the system ideal to study invasion through bone marrow or other HA-rich connective tissues. The bio-compatible cross-linking process we used allows for direct encapsulation of cancer cells within the gel where they adopt a distinct, cluster-like morphology. Metastatic PCa cells in these hydrogels develop fingerlike structures, “invadopodia”, consistent with their invasive properties. The number of invadopodia, as well as cluster size, shape, and convergence, can provide a quantifiable measure of invasive potential. Among candidate hyaluronan interacting proteins that could be responsible for the behavior we observed, we found that culture in the HA hydrogel triggers invasive PCa cells to differentially express and localize receptor for hyaluronan mediated motility (RHAMM)/CD168 which, in the absence of CD44, appears to contribute to PCa motility and invasion by interacting with the HA hydrogel components. PCa cell invasion through the HA hydrogel also was found to depend on the activity of hyaluronidases. Studies shown here reveal that while hyaluronidase activity is necessary for invadopodia and inter-connecting cluster formation, activity alone is not sufficient for acquisition of invasiveness to occur. We therefore suggest that development of invasive behavior in 3D HA-based systems requires development of additional cellular features, such as activation of motility associated pathways that regulate formation of invadopodia. Thus, we report development of a 3D system amenable to dissection of biological processes associated with cancer cell motility through HA-rich connective tissues.
Experimental evidence suggested the existence of unidentified leprosy susceptibility loci in the human leukocyte antigen (HLA) complex. To identify such genetic risk factors, a high-density association scan of a 1.9-mega-base (Mb) region in the HLA complex was performed. Among 682 single-nucleotide polymorphisms (SNPs), 59 were associated with leprosy (P <.01) in 198 Vietnamese single-case leprosy families. Genotyping of these SNPs in an independent sample of 292 Vietnamese single-case leprosy families replicated the association of 12 SNPs (P <.01). Multivariate analysis of these 12 SNPs showed that the association information could be captured by 2 intergenic HLA class I region SNPs (P = 9.4 × 10−9)—rs2394885 and rs2922997 (marginal multivariate P = 2.1 × 10−7 and P = .0016, respectively). SNP rs2394885 tagged the HLA-C*15:05 allele in the Vietnamese population. The identical associations were validated in a third sample of 364 patients with leprosy and 371 control subjects from North India. These results implicated class I alleles in leprosy pathogenesis.
In 18 male healthy subjects, artesunate (200 mg)-azithromycin (1,500 mg) daily for 3 days was found to be well tolerated, with only mild gastrointestinal disturbances reported. The pharmacokinetic properties of artesunate-azithromycin given in combination are comparable to those of the drugs given alone. Artesunate and its major active metabolite, dihydroartemisinin, are responsible for most of the ex vivo antimalarial activity, with a delayed contribution by azithromycin.
Evolutionary systems biology aims to uncover the general trends and principles governing the evolution of biological networks. An essential part of this process is the reconstruction and analysis of the evolutionary histories of these complex, dynamic networks. Unfortunately, the methodologies for representing and exploiting such complex evolutionary histories in large scale studies are currently limited. Here, we propose a new formalism, called EvoluCode (Evolutionary barCode), which allows the integration of different evolutionary parameters (eg, sequence conservation, orthology, synteny …) in a unifying format and facilitates the multilevel analysis and visualization of complex evolutionary histories at the genome scale. The advantages of the approach are demonstrated by constructing barcodes representing the evolution of the complete human proteome. Two large-scale studies are then described: (i) the mapping and visualization of the barcodes on the human chromosomes and (ii) automatic clustering of the barcodes to highlight protein subsets sharing similar evolutionary histories and their functional analysis. The methodologies developed here open the way to the efficient application of other data mining and knowledge extraction techniques in evolutionary systems biology studies. A database containing all EvoluCode data is available at: http://lbgi.igbmc.fr/barcodes.
systems biology; evolutionary history; multilevel data analysis; data representation; data visualization; data mining
To evaluate the clinical and functional results of the surgical treatment of bifid thumb type IV in children.
Materials and methods
A retrospective study was undertaken from January 1995 to December 2006. Clinical and radiographic evaluations were made according to Wassel’s classification. The patients were performed by transferring an epiphyseal segment of the proximal phalanx with insertion of the abductor pollicis brevis tendon into the radial side of the epiphyseal proximal phalanx of the ulnar thumb. All patients were operated using one of five surgical procedures for bicephalous metacarpus, cartilaginous connection between the radial and ulnar proximal phalanges, the angular deformity of the metacarpophalangeal joint (MPJ) is >20°, and zigzag deformities. The postoperative results of the patients were evaluated for both function and cosmesis according to Tien’s modified Tada scoring system.
One hundred and sixty-four patients (102 females, 62 males) were included in this study. The MPJ was stable in 170 thumbs, 15 thumbs had 10° of radial instability, and new collateral ligaments were augmented in 27 thumbs. The alignment was normal in 75 thumbs, with alignment of the interphalangeal joint (IPJ) in 101 thumbs and alignment of the MPJ in 75 thumbs. Postoperatively, there were zigzag deformities in four thumbs (developed zigzag in two thumbs, recurrent zigzag in two thumbs); there was no first web space in those hands. There were four of 185 thumbs with thumb stiffness. The abductor function of 185 thumbs was as follows: >70° in 158 thumbs (85.4%), 50°–70° in 21 thumbs (11.4%), and <50° in six thumbs (3.2%). At the latest follow-up evaluation, no evidence of physeal growth injury or growth arrest was observed in any patient. Overall, we attained good results in 140 thumbs (75.7%), fair results in 36 thumbs (19.4%), and poor results in nine thumbs (4.9%).
We recommend the use of an epiphyseal segment of the proximal phalanx with insertion of the abductor pollicis brevis tendon into the radial side of the epiphyseal proximal phalanx of the ulnar thumb and to restore anatomical insertion of the abductor pollicis brevis muscle. The technique is simple, safe, and effective for thumb abductor function in the treatment of bifid thumb type IV in children.
Thumb; Congenital duplication; Polydactyly; Duplicated thumb
Angiogenesis, the formation of new blood vessels from existing vasculature, plays an essential role in tumor growth, invasion, and metastasis. 16K hPRL, the antiangiogenic 16-kDa N-terminal fragment of human prolactin was shown to prevent tumor growth and metastasis by modifying tumor vessel morphology.
Here we investigated the effect of 16K hPRL on tumor vessel maturation and on the related signaling pathways. We show that 16K hPRL treatment leads, in a murine B16-F10 tumor model, to a dysfunctional tumor vasculature with reduced pericyte coverage, and disruption of the PDGF-B/PDGFR-B, Ang/Tie2, and Delta/Notch pathways. In an aortic ring assay, 16K hPRL impairs endothelial cell and pericyte outgrowth from the vascular ring. In addition, 16K hPRL prevents pericyte migration to endothelial cells. This event was independent of a direct inhibitory effect of 16K hPRL on pericyte viability, proliferation, or migration. In endothelial cell-pericyte cocultures, we found 16K hPRL to disturb Notch signaling.
Taken together, our data show that 16K hPRL impairs functional tumor neovascularization by inhibiting vessel maturation and for the first time that an endogenous antiangiogenic agent disturbs Notch signaling. These findings provide new insights into the mechanisms of 16K hPRL action and highlight its potential for use in anticancer therapy.
We present the case of a 32-year-old woman who presented to the emergency department with a witnessed cardiac arrest. She was otherwise healthy with no cardiac risk factors and had undergone an uneventful repeated cesarean section 3 days priorly. The patient underwent defibrillation, out of ventricular fibrillation to a perfusing sinus rhythm, and was taken to the catheterization laboratory where coronary angiography findings showed spontaneous dissection of the left anterior descending artery. The patient received a total of 6 stents during her hospital stay and was eventually discharged in good condition. Spontaneous coronary artery dissection is a rare entity with a predilection for pregnant or postpartum women. Early diagnosis and treatment are key for survival, and when identified early, mortality rate is reduced.
Micro-blogging services such as Twitter offer the potential to crowdsource epidemics in real-time. However, Twitter posts (‘tweets’) are often ambiguous and reactive to media trends. In order to ground user messages in epidemic response we focused on tracking reports of self-protective behaviour such as avoiding public gatherings or increased sanitation as the basis for further risk analysis.
We created guidelines for tagging self protective behaviour based on Jones and Salathé (2009)’s behaviour response survey. Applying the guidelines to a corpus of 5283 Twitter messages related to influenza like illness showed a high level of inter-annotator agreement (kappa 0.86). We employed supervised learning using unigrams, bigrams and regular expressions as features with two supervised classifiers (SVM and Naive Bayes) to classify tweets into 4 self-reported protective behaviour categories plus a self-reported diagnosis. In addition to classification performance we report moderately strong Spearman’s Rho correlation by comparing classifier output against WHO/NREVSS laboratory data for A(H1N1) in the USA during the 2009-2010 influenza season.
The study adds to evidence supporting a high degree of correlation between pre-diagnostic social media signals and diagnostic influenza case data, pointing the way towards low cost sensor networks. We believe that the signals we have modelled may be applicable to a wide range of diseases.
The microscopic observation drug susceptibility assay (MODS) is a novel and promising test for the early diagnosis of tuberculosis (TB). We evaluated the MODS assay for the early diagnosis of TB in HIV-positive patients presenting to Pham Ngoc Thach Hospital for Tuberculosis and Lung Diseases in southern Vietnam. A total of 738 consecutive sputum samples collected from 307 HIV-positive individuals suspected of TB were tested by smear, MODS, and the mycobacteria growth indicator tube method (MGIT). The diagnostic sensitivity and specificity of MODS compared to the microbiological gold standard (either smear or MGIT) were 87 and 93%, respectively. The sensitivities of smear, MODS, and MGIT were 57, 71, and 75%, respectively, against clinical gold standard (MODS versus smear, P < 0.001; MODS versus MGIT, P = 0.03). The clinical gold standard was defined as patients who had a clinical examination and treatment consistent with TB, with or without microbiological confirmation. For the diagnosis of smear-negative patients, the sensitivities of MODS and MGIT were 38 and 45%, respectively (P = 0.08). The median times to detection using MODS and MGIT were 8 and 11 days, respectively, and they were 11 and 17 days, respectively, for smear-negative samples. The original bacterial/fungal contamination rate of MODS was 1.1%, while it was 2.6% for MGIT. The cross-contamination rate of MODS was 4.7%. In conclusion, MODS is a sensitive, specific, and rapid test that is appropriate for the detection of HIV-associated TB; its cost and ease of use make it particularly useful in resource-limited settings.