Rivaroxaban and apixaban induce clotting factor Xa fibrinolytic activity.

Essentials Activated clotting factor X (FXa) acquires fibrinolytic cofactor function after cleavage by plasmin. FXa-mediated plasma fibrinolysis is enabled by active site modification blocking a second cleavage. FXa-directed oral anticoagulants (DOACs) alter FXa cleavage by plasmin. DOACs enhance FX-dependent fibrinolysis and plasmin generation by tissue plasminogen activator. BACKGROUND: When bound to an anionic phospholipid-containing membrane, activated clotting factor X (FXa) is sequentially cleaved by plasmin from the intact form, FXaα, to FXaß and then to Xa33/13. Tissue-type plasminogen activator (t-PA) produces plasmin and is the initiator of fibrinolysis. Both FXaß and Xa33/13 enhance t-PA-mediated plasminogen activation. Although stable in experiments using purified proteins, Xa33/13 rapidly loses t-PA cofactor function in plasma. Bypassing this inhibition, covalent modification of the FXaα active site prevents Xa33/13 formation by plasmin, and the persistent FXaß enhances plasma fibrinolysis. As the direct oral anticoagulants (DOACs) rivaroxaban and apixaban bind to the FXa active site, we hypothesized that they similarly modulate FXa fibrinolytic function. METHODS: DOAC effects on fibrinolysis and the t-PA cofactor function of FXa were studied in patient plasma, normal pooled plasma and purified protein experiments by the use of light scattering, chromogenic assays, and immunoblots. RESULTS: The plasma of patients taking rivaroxaban showed enhanced fibrinolysis correlating with FXaß. In normal pooled plasma, the addition of rivaroxaban or apixaban also shortened fibrinolysis times. This was related to the cleavage product, FXaß, which increased plasmin production by t-PA. It was confirmed that these results were not caused by DOACs affecting activated FXIII-mediated fibrin crosslinking, clot ultrastructure and thrombin-activatable fibrinolysis inhibitor activation in plasma. CONCLUSION: The current study suggests a previously unknown effect of DOACs on FXa in addition to their well-documented anticoagulant role. By enabling the t-PA cofactor function of FXaß in plasma, DOACs also enhance fibrinolysis. This effect may broaden their therapeutic indications.

ASXL2 promotes proliferation of breast cancer cells by linking ERα to histone methylation.

Estrogen receptor alpha (ERα) has a pivotal role in breast carcinogenesis by associating with various cellular factors. Selective expression of additional sex comb-like 2 (ASXL2) in ERα-positive breast cancer cells prompted us to investigate its role in chromatin modification required for ERα activation and breast carcinogenesis. Here, we observed that ASXL2 interacts with ligand E2-bound ERα and mediates ERα activation. Chromatin immunoprecipitation-sequencing analysis supports a positive role of ASXL2 at ERα target gene promoters. ASXL2 forms a complex with histone methylation modifiers including LSD1, UTX and MLL2, which all are recruited to the E2-responsive genes via ASXL2 and regulate methylations at histone H3 lysine 4, 9 and 27. The preferential binding of the PHD finger of ASXL2 to the dimethylated H3 lysine 4 may account for its requirement for ERα activation. On ASXL2 depletion, the proliferative potential of MCF7 cells and tumor size of xenograft mice decreased. Together with our finding on the higher ASXL2 expression in ERα-positive patients, we propose that ASXL2 could be a novel prognostic marker in breast cancer.

The role of each compartment in a two-compartment vertical flow reactor for ferruginous mine water treatment.

A vertical flow reactor (VFR) has been suggested for remediation of ferruginous mine drainage that passes down through an accreting bed of ochre. However, a VFR has a limited operation time until the system begins to overflow. In this study, a mathematical model was developed as a part of the effort to explore the operation of a VFR, showing dynamic changes in the head differences, ochre depths, and Fe(II)/Fe(III) concentrations in the effluent flow. The analysis showed that VFR operation time extended from 148.5 days to 163 days in an equally divided and to 168.4 days in asymmetrically (0.72:0.28) divided two-compartment VFR, suggesting that an optimum compartment ratio exists that maximizes the VFR operation time. A constant head filtration in the first compartment maximized filtration efficiency and thus prolonged VFR longevity in the two-compartment VFR. Fe(II) oxidation and ochre formation should be balanced with the permeability of the ochre bed to maximize the VFR operation time and minimize the residual Fe(II) in the effluent. Accelerated Fe(II) oxidation affected the optimum ratio of the compartment area and reduced the residual Fe(II) in the effluent. The VFR operation time can be prolonged significantly from 764 days to 3620 days by increasing the rate of ochre formation, much more than by accelerating the Fe(II) oxidation. During the prolonged VFR operation, ochre formed largely in the first compartment, while overflowing mine water with reduced iron content was effectively filtered in the second compartment. These results not only provide a better understanding of VFR operation but also suggest the direction of evolution of two-compartment VFR toward a compact and highly efficient facility integrated with an aerated cascade and with automatic coagulant feeding.

Using combination therapy to override stromal-mediated chemoresistance in mutant FLT3-positive AML: synergism between FLT3 inhibitors, dasatinib/multi-targeted inhibitors and JAK inhibitors.

Acute myeloid leukemia (AML) progenitors are frequently characterized by activating mutations in the receptor tyrosine kinase Fms-like tyrosine kinase-3 (FLT3). Protein tyrosine kinases are integral components of signaling cascades that have a role in both FLT3-mediated transformation as well as viability pathways that are advantageous to leukemic cell survival. The bone marrow microenvironment can diminish AML sensitivity to tyrosine kinase inhibitors. We hypothesized that inhibition of protein kinases in addition to FLT3 may be effective in overriding drug resistance in AML. We used a cell-based model mimicking stromal protection as part of an unbiased high-throughput chemical screen to identify kinase inhibitors with the potential to override microenvironment-mediated drug resistance in mutant FLT3-positive AML. Several related multi-targeted kinase inhibitors, including dasatinib, with the capability of reversing microenvironment-induced resistance to FLT3 inhibition were identified and validated. We validated synergy in vitro and demonstrated effective combination potential in vivo. In particular Janus kinase inhibitors were effective in overriding stromal protection and potentiating FLT3 inhibition in primary AML and cell lines. These results hint at a novel concept of using combination therapy to override drug resistance in mutant FLT3-positive AML in the bone marrow niche and suppress or eradicate residual disease.

Comparison between thymoglobulin and ATGAM as an induction agent in adult kidney transplantation: a single-center experience.

OBJECTIVE: The best antithymocyte globulin (ATG) preparation for induction suppression in kidney transplant recipients is still not clear. The aim of this study was to identify short- and long-term outcomes in kidney transplant recipients who received thymoglobulin or ATGAM as an induction agent. METHODS: We retrospectively reviewed patients who underwent kidney transplantation from 1996 to 2010. Recipients were classified according to the ATG preparation. RESULTS: One hundred fifty-two patients (64.4%) received thymoglobulin and 84 (35.6%) received ATGAM. The occurrence of delayed graft function in patients receiving thymoglobulin was higher than in patients receiving ATGAM (P = .005), but serum creatinine levels and acute rejection after kidney transplantation were not different between the two groups. The death-censored graft survival curve in thymoglobulin recipients was higher than in ATGAM recipients (P = .027). Bacterial infection was a predisposing factor for graft survival (P = .008). CONCLUSION: The efficacy of thymoglobulin induction is generally better than that of ATGAM induction, and prevention of bacterial infections was just as important as the use of ATG because bacterial infection was an important risk factor for graft failure.

Inactivated H9N2 avian influenza virus vaccine with gel-primed and mineral oil-boosted regimen could produce improved immune response in broiler breeders.

The frequent economic losses incurred with H9N2 low pathogenic avian influenza viruses (LPAI) infection have raised serious concerns for the poultry industry. A 1-dose regimen with inactivated H9N2 LPAI vaccine could not prevent vaccinated poultry from becoming infected and from shedding wild viruses. A study was conducted to determine whether a 2-dose regimen of inactivated H9N2 LPAI vaccine could enhance the immunologic response in chickens. Such gel-primed and mineral oil-boosted regimen has produced encouraging results associated with improved immune responses to an H9N2 LPAI. This strategy could be cost effective and helpful for preventing avian influenza virus in the poultry industry.

A novel method for air drying aloe leaf slices by covering with filter papers as a shrink-proof layer.

To prevent the shrinkage of aloe vera slices during air drying, a method utilizing a shrink-proof layer was developed. The sample was configured of whole leaf aloe slices, where 1 side or both sides were covered with filter papers as shrink-proof layers. After air drying by varying the air temperature and the slice thickness, the drying characteristics, as well as several quality factors of the dried aloe vera leaf slices, were analyzed. In the simulation of the drying curves, the modified Page model showed the best fitness, representing a diffusion-controlled drying mechanism. Nonetheless, there was a trace of a constant-rate drying period in the samples dried by the method. Shrinkage was greatly reduced, and the rehydration ratios increased by approximately 50%. Scanning electron microscopic analysis revealed that the surface structure of original fibrous form was well sustained. FT-IR characteristics showed that the dried samples could sustain aloe polysaccharide acetylation. Furthermore, the functional properties of the dried slices including water holding capacity, swelling, and fat absorption capability were improved, and polysaccharide retention levels increased by 20% to 30%. Therefore, we concluded that application of shrink-proof layers on aloe slices provides a novel way to overcome the shrinkage problems commonly found in air drying, thereby improving their functional properties with less cost. Practical Application: This research article demonstrates a novel air drying method using shrink-proof layers to prevent the shrinkage of aloe slices. We analyzed extensively the characteristics of shrinkage mechanism and physical properties of aloe flesh gels in this drying system. We concluded that this method can be a beneficial means to retain the functional properties of dried aloe, and a potential alternative to freeze drying, which is still costly.

Immunoprophylactic effect of chicken egg yolk immunoglobulin (Ig Y) against porcine epidemic diarrhea virus (PEDV) in piglets.

Porcine epidemic diarrhea virus (PEDV) is the causative agent of neonatal diarrhea in piglets, which causes high mortality rates. In this study, the immunoprophylactic effects of chicken egg yolk immunoglobulin (Ig Y) against PEDV were investigated in neonatal pigs. Ig Y was found to reduce the mortality in piglets after challenge exposures. The field application of Ig Y also revealed significant differences in survival rates of piglets given Ig Y, as compared with placebo or control. The results in this study indicated that Ig Y against PEDV could be an alternative way of supplementing prophylactic measures like colostral antibodies from sows.

Adenoid cystic carcinoma of the maxillary antrum.

PURPOSE: To investigate characteristic clinical features and outcome for patients with adenoid cystic carcinoma (ACC) of the maxillary antrum. PATIENTS AND METHODS: Twenty-two patients with ACC of the maxillary antrum were initially treated with surgery alone (3 patients), radiation alone (9 patients), or a combination of surgery and radiation (10 patients). Salvage treatment for initial failure was individualized. Patterns of failure, survival, and prognostic factors were retrospectively analyzed. RESULTS: The most frequent site of failure was local recurrence at the primary site (72.7%). All patients treated with either surgery alone or radiation alone experienced one or more local recurrences, whereas patients who received planned combined surgery and radiation had a much lower local recurrence rate (40%). Neck node failure (4.6%) was an uncommon event, whereas distant metastases were clinically documented in seven patients (32%). Most of the treatment failures appeared within 5 years, but treatment failures after 5 years were not uncommon. The overall survival and disease-free survival rates at 10 years were 37.6% and 13.6%, respectively. Clinicopathological factors, such as location of primary tumor, tumor stage, and histological grade were of no value in predicting a favorable survival. The significant prognostic factors influencing 10-year survival were the pathological finding of perineural invasion and the initial mode of treatment. CONCLUSION: ACC of the maxillary antrum represented a unique natural history characterized by a more aggressive tumor behavior and an unfavorable prognosis. Combined surgery and radiotherapy is recommended for optimal local control and survival.

The alterations of the activities of coagulation inhibitors and fibrinolytic factors in stored cord blood could affect the yield of progenitor cells during processing.

We assessed the changes in the activities of hemostatic variables by the storage temperature and time interval between collection and separation of cord blood (CB) and analyzed their relationship with the yield of progenitor cells during processing. Total nucleated cell (TNC) and CD34+ cell counts were significantly higher in the CB stored at ambient temperature than at 4 degrees C. The significant loss of TNC and CD34+ cells continued to 24 h after collection in CB stored at 4 degrees C, but loss of TNC began only after 24 h at ambient temperature. There were no changes in the plasma activities of antithrombin III (ATIII) and plasminogen. The activity of protein C was decreased significantly until 24 h after collection, particularly in CB stored at 4 degrees C. The activity of alpha2-antiplasmin was decreased until 24 h in CB stored at 4 degrees C and from 24 h in CB stored at ambient temperature. These data suggest that the alterations in the activities of coagulation inhibitors and fibrinolytic factors could be an important factor in coagulability, particularly in CB stored at 4 degrees C compared to ambient temperature, and also affect the yield of progenitor cells in processed CB.

Involvement of heat shock factor in regulating transcriptional activation of MDR1 gene in multidrug-resistant cells.

The present study provides evidence that heat shock factor (HSF) may be involved in a transacting factor modulating multidrug resistance 1 (MDR1) gene. In conjunction with the presence of several heat shock elements (HSEs) in the 5' region of the MDR1 gene, we compared the level of HSF which binds to HSEs in multidrug-resistant P388/M and FM3A/M cells with that in their parental counterparts. Under unstressed condition, these multidrug-resistant cells showed constitutive HSF DNA-binding activity in the nucleus of the cells, whereas their parental counterparts did not show detectable HSF DNA-binding activity. We found that H-87, protein kinase A inhibitor, inhibited HSF DNA-binding activity in heat-shocked P388/M cells and also suppressed the levels of hsp90 and hsp70. These results demonstrated that HSF might be an important transcriptional regulator for inducing MDR1 gene, and modulation of HSF activity might be a useful potential target for reversing MDR.

Radiosensitization of hypoxic tumor cells in vitro by nitric oxide.

PURPOSE: The effects of nitric oxide (NO) on the radiosensitivity of SCK tumor cells in oxic and hypoxic environments in vitro were studied. METHODS AND MATERIALS: NO was delivered to cell suspensions using the NO donors 2,2-diethyl-1-nitroso-oxyhydrazine sodium salt (DEA/NO), and a spermine/nitric oxide complex (SPER/NO), which release NO at half-lives of 2.1 min and 39 min at pH 7.4, respectively. The cells were suspended in media containing DEA/NO or SPER/NO for varying lengths of time under oxic or hypoxic conditions, irradiated, and the clonogenicity determined. RESULTS: Both compounds markedly radiosensitized the hypoxic cells. The drug enhancement ratios (DER) for 0.1, 1.0, and 2.0 mM DEA/NO were 2.0, 2.3 and 3.0, respectively, and those for 0.1, 1.0, and 2.0 mM SPER/NO were 1.6, 2.3, and 2.8, respectively. Aerobic cells were not radiosensitized by DEA/NO or SPER/NO. When DEA/ NO and SPER/NO were incubated in solution overnight to allow release of NO, they were found to have no radiosensitizing effect under hypoxic or oxic conditions indicating the sensitization by the NO donors was due to the NO molecule released from these drugs. At the higher concentrations, SPER/NO was found to be cytotoxic in aerobic conditions but not in hypoxic conditions. DEA/NO was only slightly toxic to the cells in both aerobic and hypoxic conditions. CONCLUSIONS: NO released from NO donors DEA/NO and SPER/NO is as effective as oxygen to radiosensitize hypoxic cells in vitro. Its application to the radiosensitization of hypoxic cells in solid tumors remains to be investigated.

Tumour pO2 can be increased markedly by mild hyperthermia.

We have studied the feasibility of improving tumour oxygenation with hyperthermia at modest temperatures which are achievable with the use of presently available clinical hyperthermia machines. FSaII tumours grown s.c. in the leg of C3H mice and R3230 AC tumours grown s.c. in the leg of Fischer rats were heated with a water bath and the tumour pO2 was determined with an Eppendorf pO2 histograph. The median pO2 in 7-8 mm diameter control FSaII tumours was 6.5 +/- 0.5 mmHg and it increased to 16.6 +/- 1.1 mmHg when the tumours were heated at 41.5 degrees C for 1 h. The median pO2 in 10 mm diameter control R3230 AC tumours was 3.7 +/- 0.3 mmHg. Heating at 42.5 degrees C for 30 min increased the median pO2 in the R3230 AC tumours to 12.2 +/- 1.8 mmHg. The pO2 in FSaII tumours measured 24 h after heating at 41.5 degrees C for 1 h was still higher than the pO2 before heating. The % frequency of pO2 values lower than 5 mmHg decreased markedly when the tumours were heated at the modest temperatures mentioned above. Modest temperature hyperthermia (MTH) may be an efficient and useful means to improve the oxygenation of human tumours.

Avaliaçäo da prevalência de cárie dentária em escolares de localidade urbana da regiäo sudeste do Brasil / Evaluation of the prevalence of dental caries in school-children in an urban area of Southeastern Brazil

Realizou-se no ano de 1994, na cidade de Paulínia, Estado de Säo Paulo (Brasil), um levantamento epidemiológico de cárie dentária com o intuito de comparar a atual prevalência com os dados de um estudo prévio de 1980. Foram examinados 1.416 escolares de 7 a 14 anos de idade, de ambos os sexos, por 10 dentistas previamente calibrados, utilizando-se os índices CPO.D e CPO.S. Verificou-se que houve uma queda da prevalência de cárie em 67,8 por cento em relaçäo aos dados de 1980. Observou-se uma inversäo dos componentes do índice CPO-D: em 1980 prevalecia o componente cariado (69,5 por cento), enquanto que o componente obturado prevaleceu em 1994 (79,0 por cento). Os componentes extraídos e a extraçäo indicada praticamente desapareceram no ano de 1994

Lysine production from methanol at 50 degrees C using Bacillus methanolicus: Modeling volume control, lysine concentration, and productivity using a three-phase continuous simulation.

A simulation was developed based on experimental data obtained in a 14-L reactor to predict the growth and L-lysine accumulation kinetics, and change in volume of a large-scale (250-m(3)) Bacillus methanolicus methanol-based process. Homoserine auxotrophs of B. methanolicus MGA3 are unique methylotrophs because of the ability to secrete lysine during aerobic growth and threonine starvation at 50 degrees C. Dissolved methanol (100 mM), pH, dissolved oxygen tension (0.063 atm), and threonine levels were controlled to obtain threonine-limited conditions and high-cell density (25 g dry cell weight/L) in a 14-L reactor. As a fed-batch process, the additions of neat methanol (fed on demand), threonine, and other nutrients cause the volume of the fermentation to increase and the final lysine concentration to decrease. In addition, water produced as a result of methanol metabolism contributes to the increase in the volume of the reactor. A three-phase approach was used to predict the rate of change of culture volume based on carbon dioxide production and methanol consumption. This model was used for the evaluation of volume control strategies to optimize lysine productivity. A constant volume reactor process with variable feeding and continuous removal of broth and cells (VF(cstr)) resulted in higher lysine productivity than a fed-batch process without volume control. This model predicts the variation in productivity of lysine with changes in growth and in specific lysine productivity. Simple modifications of the model allows one to investigate other high-lysine-secreting strains with different growth and lysine productivity characteristics. Strain NOA2#13A5-2 which secretes lysine and other end-products were modeled using both growth and non-growth-associated lysine productivity. A modified version of this model was used to simulate the change in culture volume of another L-lysine producing mutant (NOA2#13A52-8A66) with reduced secretion of end-products. The modified simulation indicated that growth-associated production dominates in strain NOA2#13A52-8A66. (c) 1996 John Wiley & Sons, Inc.

Prognosis and clinical relevance of recurrent oral ulceration in Behçet's disease.

There is no way of predicting whether a patient with recurrent oral ulcerations (ROU) will develop Behçet's disease (BD). In the absence of a valid laboratory test to exclude BD, such oral ulcerations result in a diagnostic problem when they occur as the sole and earliest manifestation of disease. We assessed the prognosis of ROU by performing prospective evaluations of 67 patients who had only a history of ROU and were registered at the Behçet's Disease Specialty Clinic at Severance Hospital of Yonsei University, Seoul, Korea. Thirty-five patients (52.2%) developed overt manifestations of BD at an average of 7.7 years after the onset of ROU. The frequency of recurrence was 9.8 times per year in progressive cases. From these results, it appears that highly recurrent ROU is a warning signal for BD. Careful examinations of patients, including their minor symptoms, additional laboratory tests, and regular follow-ups by physicians are required for proper diagnosis.

Ichthyosis bullosa of Siemens: report of a sporadic case.

We report a sporadic case of ichthyosis bullosa of Siemens occurring in a Korean boy. In this report, the varied findings of the clinical features in one subject over five years are presented along with an investigation of the ultrastructural alteration. The patient had suffered from blistering, superficial peeling, and dark-grey colored lichenified patches on the extremities since infancy. As he grew older, the lesions were more localized to the elbows, knees, buttock and the dorsal aspects of the hands and feet, and were replaced by yellowish, lichenified plaques. Since the original report of Siemens in 1937, nine families including one sporadic case have been reported in the literature. To our knowledge, this is the second report of sporadic case of IBS.

Acral Darier's disease: report of a case complicated by Kaposi's varicelliform eruption.

We report a case of acral Darier's disease in a 16-year-old Korean boy that was complicated by Kaposi's varicelliform eruption. It is uncommon in Darier's disease for dyskeratotic papules to be limited to acral areas. In our patient, Kaposi's varicelliform eruption occurred on the face where there were no preexisting lesions of Darier's disease.

Cutaneous reaction induced by retained bee stinger.

The clinical features and histopathologic findings of a 54-year-old Korean male who had retained the sting apparatus of a bee for four months are described. The clinical features showed ulcerative, erythematous plaques with irregular borders which resembled cutaneous neoplasms such as squamous cell carcinoma. Histopathologic findings included epidermal necrosis and marked pseudoepitheliomatous hyperplasia. In the dermis, the stick-shaped sting apparatus of the bee was demonstrated and intense lymphohistiocytic and eosinophilic infiltrations were noted.