Chondroitin sulfate liposome: clustering toward high functional efficiency.

Chondroitin sulfate (CS) is a linear polysaccharide chain of alternating residues of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc), modified with sulfate groups. Based on the structure, CS chains bind to bioactive molecules specifically and regulate their functions. For example, CS whose GalNAc is sulfated at the C4 position, termed CSA, and CS whose GalNAc is sulfated at both C4 and C6 positions, termed CSE, bind to a malaria protein VAR2CSA and receptor type of protein tyrosine phosphatase sigma (RPTPσ), respectively, in a specific manner. Here, we modified CSA and CSE chains with phosphatidylethanolamine (PE) at a reducing end, attached them to liposomes containing phospholipids and generated CSA and CSE liposomes. The CS-PE was incorporated into the liposome particles efficiently. Inhibition ELISA revealed specific interaction of CSA and CSE with recombinant VAR2CSA and RPTPσ, respectively, more efficiently than CS chains alone. Furthermore, CSE liposome was specifically incorporated into RPTPσ-expressing HEK293T cells. These results indicate CS liposome as a novel and efficient drug delivery system, especially for CS-binding molecules.

Regional variations and sex-related differences of stiffness in human tracheal ligaments.

PURPOSE: There have been numerous studies focused on the stiffness of tracheal cartilage. However, no research has been conducted specifically on the annular ligament, nor have any regional differences in the annular ligament been identified. The purpose of this study was to investigate the stiffness of the ligaments present between the thyroid, cricoid and tracheal cartilages. METHODS: The ligaments were identified in the cervical region of living subjects with ultrasonography. The stiffness of the ligaments was measured from the body surface using a digital palpation device (MyotonPRO). Since it is impossible to measure the entire trachea in a living subject, an additional measurement was performed on human cadavers. RESULTS: Both in vivo and cadaveric investigations found that the stiffness of annular ligaments decreased gradually from the superior to inferior parts. There was no difference in the stiffness between males and females in the superior part of the trachea. However, the stiffness of the middle and inferior parts was predominantly higher in females than in males. Furthermore, males showed significant differences in stiffness between the superior and middle parts, while females showed no significant differences. CONCLUSION: These results reveal that there are regional and sex-related differences in the stiffness of human tracheal ligaments.

Assessing site-specificity of the biomechanical properties of hamstring aponeuroses using MyotonPRO: A cadaveric study.

BACKGROUND: Hamstring muscles are the most frequently reported sites of muscle strain injuries, especially near the bi-articular muscles' myotendinous junction, where aponeurosis provides a connective tissue network linking muscle fibers to the tendon. This study aimed to investigate the reliability and site-specific differences of hamstring aponeuroses under different conditions (formalin and urea) using MyotonPRO. METHODS: Eight hamstring muscle groups were dissected from four human cadavers (two males and two females) aged 83-93 years. Measurements of the mechanical properties of the aponeuroses from the superficial and deep regions of biceps femoris long head, semitendinosus, and semimembranosus (after formalin solution immersion) were done using MyotonPRO (intra-rater reliability was examined within a 24-h interval), following which the hamstring aponeuroses were measured using a similar procedure after urea solution immersion. FINDINGS: Test-retest (intra-rater) results revealed that the MyotonPRO measurement of tone, stiffness, relaxation, and creep of cadaveric aponeuroses presented good to excellent reliability (ICC: 0.86 to 0.98). There were no significant differences in tone, stiffness, elasticity, relaxation, and creep among the six sites of hamstring aponeuroses under both formalin and urea conditions. Significant differences between formalin and urea conditions were found in the tone, stiffness, relaxation, and creep of hamstring aponeuroses (P < 0.05). INTERPRETATION: These results suggested that the biomechanical properties of hamstring aponeuroses showed homogeneity between the sites using MyotonPRO. Urea solution could potentially neutralize the effect of formalin on the biomechanical properties of cadaveric muscle-aponeurosis-tendon units. The present findings might influence the design of subsequent cadaveric studies on hamstring muscle strains.

Biomechanical assessment of gastrocnemii and Achilles tendon using MyotonPRO: in vivo measurements, and preliminary in situ measurements using formalin-fixed tissues.

PURPOSE: This study aims to evaluate the reliability and validity of using MyotonPRO to quantify the mechanical properties of the muscle-tendon unit through in vivo measurements and preliminary in situ measurements using formalin-fixed tissues. MATERIALS AND METHODS: The mechanical properties of gastrocnemii and the Achilles tendon of 12 healthy adults (six males and six females, 34.9 ± 5.8 years) were examined for in vivo test twice within a day and once post-24 hours using MyotonPRO, while nine human cadavers (formalin-fixed, 3 males and 6 females, 89.9 ± 5.1 years) were assessed for preliminary in situ test with identical time schedule to evaluate the within-day and inter-day reliability and validity. RESULTS: In vivo tests had very high within-day (ICC: 0.96-0.99) and inter-day reliability (ICC: 0.83-0.96), while in situ tests (formalin-fixed tissues) showed high within-day (ICC: 0.87-0.99) and inter-day reliability (ICC: 0.76-0.98) for the results of tone and stiffness. There was no significant difference in the stiffness of the free part of the Achilles tendon between in vivo and in situ conditions. The stiffness of the lateral gastrocnemius (r = 0.55, p = 0.018), proximal part of the Achilles tendon (r = 0.56, p = 0.015), and free part of the Achilles tendon (r = 0.47, p = 0.048) before removing the skin was significantly correlated with that after removing the skin condition. CONCLUSIONS: The findings of the current study suggest that MyotonPRO is reliable and valid for evaluating tendon stiffness both in vivo and in situ (formalin-fixed tissues).

Discordance of global assessment between the patients and physicians predicts 9-year pain-related outcomes in rheumatoid arthritis patients.

Introduction: Perspectives regarding the disease state often differ between patients with rheumatoid arthritis (RA) and physicians. The aim of the present longitudinal cohort study was to investigate the impact of the discordance in global assessments between patients and physicians on 9-year pain-related outcomes in patients with rheumatoid arthritis. Method: Sixty-eight consecutive outpatients with rheumatoid arthritis on their first visit to a tertiary center were included. Baseline measurements included demographic data, drugs used, disease activity, and a modified Health Assessment Questionnaire (mHAQ). Discordance in global assessment between patients and physicians at baseline was defined as 10 mm higher in the patient global assessment (PGA) than in the physician global assessment. A 9-year follow-up assessment included pain intensity, the European Quality of Life 5 Dimensions 3 Level (EQ-5D-3L) scale, Pain Catastrophizing Scale (PCS), Hospital Anxiety and Depression Scale (HADS), Pain Disability Assessment Scale (PDAS), and Pain Self-Efficacy Questionnaire (PSEQ). Results: The number of patients with discordance was 26 (38%) in 68 patients. Patients with a 10 mm higher PGA than the physician global assessment at baseline measurements had significantly worse pain intensity, PCS score, PSEQ score, and EQ-5D-3L score measurements at the 9-year follow-up than those with concordance. A higher mHAQ score and 10 mm higher PGA at baseline were significantly independently associated with the EQ-5D-3L scale score and pain intensity at the 9-year follow-up. Conclusion: This longitudinal cohort study suggested that discordance in global assessment between patients and physicians modestly predicted worse 9-year pain-related outcomes in patients with rheumatoid arthritis.

The clinical characteristics, treatment, and survival of portopulmonary hypertension in Japan.

BACKGROUND: Portopulmonary hypertension (PoPH) refers to the simultaneous presentation of pulmonary arterial and portal hypertension. However, few reports have included the characteristics and treatments for patients with PoPH of Asian population; thus, we investigated the clinical characteristics, treatment, and survival of these patients in a Japanese cohort. METHODS: Pulmonary arterial hypertension (PAH) has been included in the National Research Project on Intractable Disease in Japan; therefore, we extracted data of patients with PoPH from the forms of newly registered cases of the project from 2012 to 2013 (for 2 years), and updated cases of the project in 2013 (Study 1, n = 36 newly registered forms, n = 46 updated forms). Additionally, for Study 2, we performed a retrospective, observational cohort study at Chiba University Hospital (n = 11). We compared the characteristics between patients with PoPH and those with idiopathic/heritable PAH (I/H-PAH). RESULTS: Both studies showed higher cardiac outputs (COs) and cardiac indexes (CIs), lower pulmonary vascular resistance (PVR), and less treated with combination therapy in patients with PoPH than those with I/H-PAH. In Study 2, the overall and disease-specific survival between PoPH and I/H-PAH were similar. Conversely, many patients (45%) had to change their PAH-specific medicine because of adverse effects. CONCLUSION: As seen in western countries, Japanese patients with PoPH showed higher COs and CIs, better exercise tolerance, and lower PVRs than patients with I/H-PAH. Further studies are needed to improve PoPH treatments.

[Improvement of the Non-aqueous Titration Method in the Japanese Pharmacopoeia for a Purpose of Advancement in Experimental Safety Using Alternative Solvents].

In this study, we attempted to improve the non-aqueous titration method using N,N-dimethylformamide (DMF) in the Japanese Pharmacopoeia seventeenth edition (JP XVII) for advancement in experimental safety. As an alternative solvent for DMF, we demonstrate titrations using dimethyl sulfoxide (DMSO), which has similar properties as and much higher safety than DMF. Five drugs (i.e., acetohexamide, glibenclamide, sulfamethoxazole, tranilast, and furosemide) listed in JP XVII use DMF as a solvent for titrations with sodium hydroxide standard solution. For these drugs, we examined whether DMF can be replaced with DMSO in quantitative analyses. As a result, a quantification similar to that of the Pharmacopoeia protocol is possible by simply replacing DMF with DMSO or using a mixed solvent of DMSO and water.

Fabrication of Naftopidil-Loaded Tablets Using a Semisolid Extrusion-Type 3D Printer and the Characteristics of the Printed Hydrogel and Resulting Tablets.

The production of three-dimensional (3D)-printed drugs holds promise for future personalized medicine. Here, we prepared tablets containing naftopidil as a model drug using a semisolid extrusion-type 3D bioprinter applicable for tissue engineering. A hydrogel is typically used as the printer ink for 3D bioprinters, and we incorporated various amounts of hydroxypropyl methylcellulose hydrogel (30%, 40%, and 50% gel) into the printer ink. The resulting 3D-printed gel product was dried to obtain tablets. The rheological properties of the printer ink changed as its composition was changed, and tablets were prepared successfully from several formulations. Increasing the amount of hydroxypropyl methylcellulose hydrogel in the printer ink led to delayed drug dissolution, decreased weight, and decreased hardness of the tablets. Delayed drug dissolution was also observed when the amount of disintegrating agent typically used in powder compression tablets was increased in the ink, and increasing the incorporated amount of the disintegrating agent crospovidone increased the hardness of the tablets. Our results will provide useful information for the preparation of tablets using semisolid extrusion-type 3D printers.

Endothelial cell-related autophagic pathways in Sugen/hypoxia-exposed pulmonary arterial hypertensive rats.

Pulmonary arterial hypertension (PAH) is characterized by progressive obstructive remodeling of pulmonary arteries. However, no reports have described the causative role of the autophagic pathway in pulmonary vascular endothelial cell (EC) alterations associated with PAH. This study investigated the time-dependent role of the autophagic pathway in pulmonary vascular ECs and pulmonary vascular EC kinesis in a severe PAH rat model (Sugen/hypoxia rat) and evaluated whether timely induction of the autophagic pathway by rapamycin improves PAH. Hemodynamic and histological examinations as well as flow cytometry of pulmonary vascular EC-related autophagic pathways and pulmonary vascular EC kinetics in lung cell suspensions were performed. The time-dependent and therapeutic effects of rapamycin on the autophagic pathway were also assessed. Sugen/hypoxia rats treated with the vascular endothelial growth factor receptor blocker SU5416 showed increased right ventricular systolic pressure (RVSP) and numbers of obstructive vessels due to increased pulmonary vascular remodeling. The expression of the autophagic marker LC3 in ECs also changed in a time-dependent manner, in parallel with proliferation and apoptotic markers as assessed by flow cytometry. These results suggest the presence of cross talk between pulmonary vascular remodeling and the autophagic pathway, especially in small vascular lesions. Moreover, treatment of Sugen/hypoxia rats with rapamycin after SU5416 injection activated the autophagic pathway and improved the balance between cell proliferation and apoptosis in pulmonary vascular ECs to reduce RVSP and pulmonary vascular remodeling. These results suggested that the autophagic pathway can suppress PAH progression and that rapamycin-dependent activation of the autophagic pathway could ameliorate PAH.

Severe Pulmonary Arteriopathy Is Associated with Persistent Hypoxemia after Pulmonary Endarterectomy in Chronic Thromboembolic Pulmonary Hypertension.

BACKGROUND: Chronic thromboembolic pulmonary hypertension (CTEPH) is characterized by occlusion of pulmonary arteries by organized chronic thrombi. Persistent hypoxemia and residual pulmonary hypertension (PH) following successful pulmonary endarterectomy (PEA) are clinically important problems; however, the underlying mechanisms remain unclear. We have previously reported that residual PH is closely related to severe pulmonary vascular remodeling and hypothesize that this arteriopathy might also be involved in impaired gas exchange. The purpose of this study was to evaluate the association between hypoxemia and pulmonary arteriopathy after PEA. METHODS AND RESULTS: Between December 2011 and November 2014, 23 CTEPH patients underwent PEA and lung biopsy. The extent of pulmonary arteriopathy was quantified pathologically in lung biopsy specimens. We then analyzed the relationship between the severity of pulmonary arteriopathy and gas exchange after PEA. We observed that the severity of pulmonary arteriopathy was negatively correlated with postoperative and follow-up PaO2 (postoperative PaO2: r = -0.73, p = 0.0004; follow-up PaO2: r = -0.66, p = 0.001), but not with preoperative PaO2 (r = -0.373, p = 0.08). Multivariate analysis revealed that the obstruction ratio and patient age were determinants of PaO2 one month after PEA (R2 = 0.651, p = 0.00009). Furthermore, the obstruction ratio and improvement of pulmonary vascular resistance were determinants of PaO2 at follow-up (R2 = 0.545, p = 0.0002). Severe pulmonary arteriopathy might increase the alveolar-arterial oxygen difference and impair diffusion capacity, resulting in hypoxemia following PEA. CONCLUSION: The severity of pulmonary arteriopathy was closely associated with postoperative and follow-up hypoxemia.

Pulmonary haemodynamics are correlated with intimal lesions in a rat model of severe PAH: attenuation of pulmonary vascular remodelling with ambrisentan.

RATIONALE: Pulmonary arterial hypertension (PAH) is characterized by obstructive lesions and vasoconstriction of the pulmonary arteries. Early therapeutic interventions with vasodilator drugs are thought to be beneficial in PAH. However, it remains unknown whether the severity of intimal obstruction is associated with increased pulmonary arterial pressure and whether reduction of vasoconstriction in the earlier stage by these drugs has a beneficial effect. Therefore, the aims of this study were to investigate these issues in a rat model of severe PAH. Methods A rat model of severe PAH was created by injection of a vascular endothelial growth factor receptor blocker in combination with hypoxia for the first 3 weeks followed by normoxia for the next 9 weeks. To assess intimal obstruction, "the pulmonary artery occlusion index (PAOI)" was developed to digitize all lesions. The small pulmonary arteries were assessed by this index, and the association between right ventricular systolic pressure (RVSP) and PAOI was investigated. An endothelin receptor antagonist, ambrisentan, was administered by gavage to rats during either hypoxia (Prevention study group, n=25) or normoxia (Early treatment group, n=15). RESULTS: PAOI showed a positive correlation with RVSP, and both RVSP and PAOI increased gradually over time. There were no severe occlusive lesions in either group, but the density of partially occlusive lesions was significantly decreased in the Prevention study group. CONCLUSION: A novel PAOI index was developed, and this index was strongly correlated with RVSP. Furthermore, ambrisentan reduced luminal occlusive lesions more effectively when treatment was given during the first 2 weeks of hypoxia.

Effects of Surgical and Medical Treatment on Quality of Life for Patients With Chronic Thromboembolic Pulmonary Hypertension.

BACKGROUND: This study aimed to investigate the predictors of quality of life (QOL) in patients with chronic thromboembolic pulmonary hypertension (CTEPH), changes in QOL after surgical and medical treatments, and the relationship between baseline QOL and survival. METHODS AND RESULTS: QOL was measured in 128 patients with CTEPH (male/female: 42/86, age: 56±12 years, surgical/medical: 65/63) using the Short-Form 36 (SF-36) questionnaire. Multiple regression analysis showed pulmonary vascular resistance (PVR) and 6-min walking distance (6MWD) were associated with physical functioning (PF) (P<0.01) and physical component summary (PCS) (P<0.01). In the surgical group, 7 subscales and 2 summary scores improved significantly, and in the medical group 6 subscales and the mental component summary, although the change in QOL was greater in the surgical group. The patients in the conventional therapy group with higher PF had significantly better survival than those with lower PF (5-years survival: 89.5% vs. 50.8%, P=0.002). This difference in survival was not observed in the group receiving pulmonary arterial hypertension (PAH)-specific therapy (100% vs. 100%, P=0.746). CONCLUSIONS: PVR and 6MWD were associated with PF or PCS in CTEPH patients. QOL improved after surgical or medical therapy, with a greater change in the surgical group. PAH-specific therapy improved survival in patients with lower PF at diagnosis.

Idiopathic bronchocentric granulomatosis in an asthmatic adolescent.

Bronchocentric granulomatosis in asthmatic patients has been generally considered to be associated with allergic bronchopulmonary aspergillosis and represent a histopathologic manifestation of fungal hypersensitivity. Here we report a case of an idiopathic bronchocentric granulomatosis in a 17-year-old man with a history of asthma. He was admitted to the hospital with a fever and cough, and a chest CT scan showed peribronchial consolidation in the pulmonary parenchyma, which was unresponsive to antibiotic therapy. The pathological findings obtained by video-assisted thoracoscopic lung biopsy revealed necrotizing granulomatous inflammation centered on bronchi and bronchioles and there was no evidence of fungal colonization, resulting in a diagnosis of idiopathic bronchocentric granulomatosis. Systemic corticosteroid therapy led to clinical and radiological recovery. Physicians should take into account the possibility of the idiopathic process in bronchocentric granulomatosis of asthmatic patients.

Association of plasma fibrinogen and plasminogen with prognosis of inoperable chronic thromboembolic pulmonary hypertension.

BACKGROUND: It is unclear whether abnormalities of coagulation or fibrinolysis are associated with disease progression of chronic thromboembolic pulmonary hypertension (CTEPH). The aim of this study was to investigate the association of these factors with the severity and prognosis of CTEPH. METHODS AND RESULTS: Between 1986 and 2011, plasma fibrinogen and plasminogen were measured in 89 of 106 consecutive patients with inoperable CTEPH (17 men; mean age, 55.9±14.1 years old; mean pulmonary arterial pressure, 44.0±12.4 mmHg) and the association of level with severity and prognosis were also examined. Seventeen patients had high fibrinogen and low plasminogen (medians, ≥291 mg/dl and <101%, respectively). These patients had significantly lower cardiac index (2.26±0.68 vs. 2.70±0.57 L·min(-1)·m(-2), P=0.007), higher pulmonary vascular resistance (PVR; 13.29±7.54 vs. 9.15±4.14 Wood units, P=0.003), and poor survival (5-year survival, 35.3% vs. 88.0%, P<0.001) compared to the other 72 patients. Additional analysis showed significantly poor survival in these patients compared with the other patients who did not have modern therapy. On multivariate analysis plasma fibrinogen, plasminogen and PVR were independent predictors of survival in medically treated patients. CONCLUSIONS: High plasma fibrinogen and low plasminogen are associated with poor survival in CTEPH patients without modern therapy.

Procalcitonin-guided antibiotic therapy in aspiration pneumonia and an assessment of the continuation of oral intake.

BACKGROUND: Procalcitonin-guided antibiotic therapy for community-acquired pneumonia is effective and safe. However, the usefulness of procalcitonin for aspiration pneumonia and its nutrition-related outcomes are unknown. METHODS: We conducted a noninferiority randomized controlled study in patients with aspiration pneumonia who were admitted to our hospital between September 2010 and January 2012. We randomly assigned 105 patients to groups with different durations of antibiotic therapy based on the procalcitonin levels upon admission (procalcitonin group) or according to the standard guidelines (control group). The primary endpoints were relapse of aspiration pneumonia and death within 30 days, with a predefined noninferiority boundary of 10%. Secondary endpoints included duration of antibiotic exposure. Furthermore, we conducted a retrospective analysis of the prognostic factors that determined continuation of oral nutritional intake, relapse of pneumonia, and in-hospital death. RESULTS: The rate of relapse and death within 30 days were similar in the procalcitonin and control groups (25% versus 37.5%; difference, -12.5%; 95% confidence interval, -30.9% to 5.9%). Procalcitonin-guided antibiotic therapy significantly shortened the median duration of antibiotic exposure (5 versus 8 days; p<0.0001); however, the continuation of oral intake was not increased (56% versus 50%; p=0.54). A multivariable analysis showed a significant association between the continuation of oral nutritional intake and the body mass index upon admission. CONCLUSIONS: Procalcitonin-guided antibiotic therapy for aspiration pneumonia can shorten the duration of antibiotic exposure, but it does not increase the continuation of oral intake (UMIN000004800).

[Retrospective analysis of pemetrexed plus cisplatin chemotherapy for elderly advanced non-small-cell lung cancer].

BACKGROUND: The efficacy of pemetrexed(PEM)plus cisplatin(CDDP)therapy for chemotherapy-naive non-squamous cell lung cancer has been reported, but the effectiveness of such a regimen for elderly patients is unknown. PURPOSE: The aim of this study is to examine the efficacy and toxicity of CDDP plus PEM therapy for elderly patients, retrospectively. METHODS: We performed a retrospective analysis of six patients 75 years old or older with non-squamous lung cancer, who underwent CDDP plus PEM therapy from June 2009 to May 2010. RESULTS: The mean age was 79. 2 years old(range, 76-82), gender: 3 males/3 females; stage: III B/IV; 1/5, pathology: all patients had adenocarcinoma without epidermal growth factor receptor (EGFR)mutation, line: first/third; 5/1. The scheduled chemotherapy of four courses was completed in four patients. The overall response rate was 50%, and the disease control rate was 83%. Grade 3/4 neutropenia and thrombocytopenia were observed in 1/2 and 1/1 patients, respectively, but no blood transfusions were needed. Severe myelosuppression was shown in patients who were impaired in renal function. Grade 3 nausea or anorexia was also observed in 50%of patients. Therefore, two patients were terminated in one courses of therapy and long-term hospitalization for them was needed. CONCLUSION: Although CDDP plus PEM therapy for elderly patients has sufficient patients compliance because of its tolerable myelosuppression, it is necessary to pay attention to deterioration in renal function and to care for nausea during chemotherapy.

[Minimal change nephrotic syndrome developing in a rheumatoid arthritis patient under etanercept treatment].

A 67-year-old female patient with rheumatoid arthritis (RA) developed minimal change nephrotic syndrome (MCNS) while under treatment with etanercept (ETN). Histopathological examination of renal biopsy specimens showed minimal-change nephropathy. Almost complete resolution of the MCNS was achieved by discontinuation of ETN and initiation of steroid therapy. There have been no reports from Japan of the occurrence of MCNS caused by ETN administration in RA patients, and only very few cases have been reported from around the world. Therefore, this case was considered to be very uncommon and worthy of reporting, and herein is a report of our patient.