Connective tissues such as tendon, ligament and cartilage are mostly composed of extracellular matrix (ECM). These tissues are insoluble, mainly due to the highly cross-linked ECM proteins such as collagens. Difficulties obtaining suitable samples for mass spectrometric analysis render the application of modern proteomic technologies difficult. Complete solubilization of them would not only elucidate protein composition of normal tissues but also reveal pathophysiology of pathological tissues. Here we report complete solubilization of human Achilles tendon and yellow ligament, which is achieved by chemical digestion combined with successive protease treatment including elastase. The digestion mixture was subjected to liquid chromatography mass spectrometry. The low specificity of elastase was overcome by accurate mass analysis achieved using FT-ICR-MS. In addition to the detailed proteome of both tissues, we also quantitatively determine the major protein composition of samples, by measuring peak area of some characteristic peptides detected in tissue samples and in purified proteins. As a result, differences between human Achilles tendon and yellow ligament were elucidated at molecular level.
Background: Immunization is one of the most cost-effective public health interventions to prevent children from contracting vaccine-preventable diseases. Indonesia launched the Expanded Program for Immunization (EPI) in 1977. However, immunization coverage remains far below the United Nations International Children's Emergency Fund (UNICEF) and World Health Organization (WHO) target of 80%. This study aims to investigate the determinants of complete immunization status among children aged 12-23 months in Indonesia. Methods: We used three waves of the Indonesian National Socioeconomic Survey (2008, 2011, and 2013) and national village censuses from the same years. Multilevel logistic regression was used to conduct the analysis. Results: The number of immunized children increased from 47.48% in 2008 to 61.83% in 2013. The presence of health professionals, having an older mother, and having more educated mothers were associated with a higher probability of a child's receiving full immunization. Increasing the numbers of hospitals, village health posts, and health workers was positively associated with children receiving full immunization. The MOR (median odds ratio) showed that children's likelihood of receiving complete immunization varied significantly among districts. Conclusions: Both household-and district-level determinants were found to be associated with childhood immunization status. Policy makers may take these determinants into account to increase immunization coverage in Indonesia.
Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) is cytologically and phenotypically heterogeneous. Retinoic acid–related orphan receptor-γt (RORγt) is a transcription factor that regulates the differentiation of naïve CD4 helper T cells to Th17 cells. In the present study, we immunohistochemically confirmed the expression of RORγt in PTCL-NOS. Pathological and clinical investigations were performed for 170 cases of PTCL-NOS. RORγt-positive cases accounted for 17.6% (30/170) of the total cases, and they showed a significantly higher frequency of CD8 positivity ( = .033), lower counts of white blood cells ( = .030) and neutrophils ( = .039) in the peripheral blood, higher levels of hypergammaglobulinemia ( = .031), a higher frequency of a complete response ( = .009), and a tendency for a lower International Prognostic Index ( = .061) and better overall survival ( = .0806). These results suggest that RORγt-positive PTCL-NOS could be a subpopulation of PTCL-NOS. Further research associated with this genomic abnormality at the transcriptional level is needed to confirm the results of this study.
An inherent difficulty in using DNA microarray technology on the early mouse embryo is its relatively small size. In this study, we investigated whether use of ES cell differentiation culture, which has no theoretical limit in the number of cells that can be generated, can improve this situation. Seven distinct ES-cell-derived populations were analyzed by DNA microarray and examined for genes whose distribution patterns are similar to those of PDGFRα, a gene implicated in differentiation of mesoderm/mesenchymal lineages. Using software developed in our laboratory, we formed a group of 30 genes which showed the highest similarity to PDGFRα, 18 of these genes were shown to be involved in development of either mesodermal, mesenchymal or neural crest cells. This list also contains several genes whose role in embryogenesis has not yet been fully identified. One such molecule is mARID3b. The mARID3b expression is found in the paraxial mesoderm and cranial mesenchyme. mARID3b-null mouse showed early embryonic lethality, and most phenotypes of this mutant appear to develop from a failure to generate a sufficient number of cranial mesenchymal cells. These results demonstrate the potential use of ES cell differentiation culture in identifying novel genes playing an indispensable role in embryogenesis.
Background Severe or life-threatening asthma exacerbation is one of the worst outcomes of asthma because of the risk of death. To date, few studies have explored the potential heterogeneity of this condition. Objectives To examine the clinical characteristics and heterogeneity of patients with severe or life-threatening asthma exacerbation. Methods This was a multicentre, prospective study of patients with severe or life-threatening asthma exacerbation and pulse oxygen saturation < 90% who were admitted to 17 institutions across Japan. Cluster analysis was performed using variables from patient-and physician-orientated structured questionnaires. Results Analysis of data from 175 patients with severe or life-threatening asthma exacerbation revealed five distinct clusters. Cluster 1 (n = 27) was younger-onset asthma with severe symptoms at baseline, including limitation of activities, a higher frequency of treatment with oral corticosteroids and short-acting beta-agonists, and a higher frequency of asthma hospitalizations in the past year. Cluster 2 (n = 35) was predominantly composed of elderly females, with the highest frequency of comorbid, chronic hyperplastic rhinosinusitis/nasal polyposis, and a long disease duration. Cluster 3 (n = 40) was allergic asthma without inhaled corticosteroid use at baseline. Patients in this cluster had a higher frequency of atopy, including allergic rhinitis and furred pet hypersensitivity, and a better prognosis during hospitalization compared with the other clusters. Cluster 4 (n = 34) was characterized by elderly males with concomitant chronic obstructive pulmonary disease (COPD). Although cluster 5 (n = 39) had very mild symptoms at baseline according to the patient questionnaires, 41% had previously been hospitalized for asthma. Conclusions & Clinical Relevance This study demonstrated that significant heterogeneity exists among patients with severe or life-threatening asthma exacerbation. Differences were observed in the severity of asthma symptoms and use of inhaled corticosteroids at baseline, and the presence of comorbid COPD. These findings may contribute to a deeper understanding and better management of this patient population.
Understanding intratumor heterogeneity is clinically important because it could cause therapeutic failure by fostering evolutionary adaptation. To this end, we profiled the genome and epigenome in multiple regions within each of nine colorectal tumors. Extensive intertumor heterogeneity is observed, from which we inferred the evolutionary history of the tumors. First, clonally shared alterations appeared, in which C>T transitions at CpG site and CpG island hypermethylation were relatively enriched. Correlation between mutation counts and patients' ages suggests that the early-acquired alterations resulted from aging. In the late phase, a parental clone was branched into numerous subclones. Known driver alterations were observed frequently in the early-acquired alterations, but rarely in the late-acquired alterations. Consistently, our computational simulation of the branching evolution suggests that extensive intratumor heterogeneity could be generated by neutral evolution. Collectively, we propose a new model of colorectal cancer evolution, which is useful for understanding and confronting this heterogeneous disease.
With the use of synthetic materials for medial patellofemoral ligament (MPFL) reconstruction, graft harvest is not necessary and this may facilitate post-operative rehabilitation. The purpose of this study was to compare the structural properties of MPFL reconstruction using a modern synthetic material (FiberTape® (FT), Arthrex) with knotless anchors or a semitendinosus (ST) tendon autograft with soft anchors. Nine human fresh-frozen amputated knees were used in this study. After the tensile strength of the native MPFL was measured, the MPFLs were reconstructed using two different surgical procedures, FT with knotless anchors (group A) and a ST with soft anchors (group B). Mechanical testing to failure of the reconstructed MPFLs was performed, and the ultimate load (N), stiffness (N/mm), and failure mode were recorded. The mean (±standard deviation) ultimate load of the native MPFL was 130.6 ± 28.7 N, and all native MPFLs failed at the femoral insertion site. Ultimate load of group A was significantly higher than that of the native MPFL (175.9 ± 34.1 N, p < 0.05). In contrast, the ultimate load of group B was significantly lower than that of the native MPFL (102.7 ± 21.4 N, p < 0.05). The mean stiffness was significantly higher for MPFLs in group A (17.4 ± 4.3 N/mm) than in group B (8.5 ± 1.8 N/mm, p < 0.05). In group A, 5 specimens failed via a knotless anchor pullout at the femoral side, 3 via pullout of knotless anchors at the patella side and 1 via fracture (cheese cut) of the femur without breakage of knotless anchor. In group B, all specimens failed via soft anchor pullout at the patella side. There was no incidence of rupture of FT or ST. FT with knotless anchors was stronger than a ST with soft tissue anchors for MPFL reconstruction.
Abstract Background The number of operations performed by a surgeon may be an indicator of surgical skill. The hand motions made by a surgeon also reflect skill and level of expertise. We hypothesized that the hand motions of expert and novice surgeons differ significantly, regardless of whether they are familiar with specific tasks during an operation. Methods This study compared 11 expert surgeons, each of whom had performed >100 laparoscopic procedures, and 27 young surgeons, each of whom had performed <15 laparoscopic procedures. Each examinee performed a specific skill assessment task, in which instrument motion was monitored using magnetic tracking system. We analyzed the paths of the centers of gravity of the tips of the needle holders and the relative paths of the tips using two mathematical methods of detrended fluctuation analysis and unstable periodic orbit analysis. Results Detrended fluctuation analysis showed that the exponent in the function describing the initial scaling exponent ( α1 ) differed significantly for experts and novices, being close to 1.0 and 1.5, respectively ( P < 0.01). This indicated that the expert group had a greater long-range coherence with an intrinsic sequence and smooth continuity among a series of motions. Likewise, unstable periodic orbit analysis showed that the second period of unstable orbit was significantly longer for experts in comparison with novices ( P < 0.01). This demonstrates mathematically that the hands of experts are more stable when performing laparoscopic procedures. Conclusions Objective evaluation of hand motion during a simulated laparoscopic procedure showed a significant difference between experts and novices.
Peripheral T cell lymphoma, not otherwise specified (PTCL-NOS), is a heterogeneous disease with respect to clinicopathological features. Cell adhesion molecule 1 (CADM1) has been reported to be ectopically expressed in adult T cell leukaemia/lymphoma (ATLL). However, the frequency of CADM1 expression remains unknown in peripheral T cell lymphomas. In the current study, CADM1 expression was analysed in 88 PTCL-NOS patients. CADM1 was expressed in 14 of 88 (15.9%) PTCL-NOS cases, and its expression was associated with C-C chemokine receptor type 4 (CCR4) expression and nuclear atypia. CADM1-positive PTCL-NOS cases (10/74) had a significantly poorer prognosis than CADM1-negative cases (64/74) (P = 0.001). Multivariate analysis confirmed that CADM1 expression was an independent prognostic factor in PTCL-NOS. These findings suggest that CADM1 expression is a novel prognostic factor for PTCL-NOS.
To assess the wrist joints of healthy volunteers in extended and loaded states versus the unloaded state by using computed tomography (CT) to analyze the 3-dimensional movements in the distal radioulnar joint (DRUJ). The dominant arms of 9 volunteers with healthy wrists were studied. We mounted a compression device onto the elbows in an inverted position. A 0-kg and 7-kg load each was applied during low-dose radiation CT imaging and a bone model was produced. We marked the insertion sites for the 4 radioulnar ligaments stabilizing the DRUJ: palmar superficial radioulnar ligament (PS-RUL), dorsal superficial radioulnar ligament (DS-RUL), dorsal deep radioulnar ligament (DD-RUL), and palmar deep radioulnar ligament (PD-RUL). Using Marai’s method, each ligament was virtualized and the length of each simulated ligament was measured. We also computed the 3-dimensional displacement and corresponding rotation of the distal ulna where it comes into contact with the radius in the sigmoid notch. The lengths of palmar ligaments (PS-RUL and PD-RUL) increased significantly under loaded conditions, and although not significant, the length of dorsal ligaments (DS-RUL and DD-RUL) tended to increase. When the wrist was loaded, the ulna rotated toward the open palmar side. The length of simulated radioulnar ligaments increased when the wrist joint was loaded in an extended position. This kinematic movement of DRUJ separation under a loading condition is different from physiological active movement. The 3-dimensional kinematic analysis revealed that palmar radioulnar ligaments were stretched during axial loading, suggesting that a tear of the palmer ligament can result from a fall on an outstretched hand.