Exogenous Nitric Oxide Suppresses in Vivo X-ray-Induced Targeted and Non-Targeted Effects in Zebrafish Embryos.

The present paper studied the X-ray-induced targeted effect in irradiated zebrafish embryos (Danio rerio), as well as a non-targeted effect in bystander naïve embryos partnered with irradiated embryos, and examined the influence of exogenous nitric oxide (NO) on these targeted and non-targeted effects. The exogenous NO was generated using an NO donor, S-nitroso-N-acetylpenicillamine (SNAP). The targeted and non-targeted effects, as well as the toxicity of the SNAP, were assessed using the number of apoptotic events in the zebrafish embryos at 24 h post fertilization (hpf) revealed through acridine orange (AO) staining. SNAP with concentrations of 20 and 100 µM were first confirmed to have no significant toxicity on zebrafish embryos. The targeted effect was mitigated in zebrafish embryos if they were pretreated with 100 µM SNAP prior to irradiation with an X-ray dose of 75 mGy but was not alleviated in zebrafish embryos if they were pretreated with 20 µM SNAP. On the other hand, the non-targeted effect was eliminated in the bystander naïve zebrafish embryos if they were pretreated with 20 or 100 µM SNAP prior to partnering with zebrafish embryos having been subjected to irradiation with an X-ray dose of 75 mGy. These findings revealed the importance of NO in the protection against damages induced by ionizing radiations or by radiation-induced bystander signals, and could have important impacts on development of advanced cancer treatment strategies.

Anti-allergic action of anti-malarial drug artesunate in experimental mast cell-mediated anaphylactic models.

BACKGROUND: Allergy is an acquired hypersensitivity reaction of the immune system mediated by cross-linking of allergen-specific IgE-bound high-affinity IgE receptors, leading to immediate mast cell degranulation. Artesunate is a semi-synthetic derivative of artemisinin, an active component of the medicinal plant Artemisia annua. Artesunate is a clinically effective anti-malarial drug and has recently been shown to attenuate allergic asthma in mouse models. This study investigated potential anti-allergic effects of artesunate in animal models of IgE-dependent anaphylaxis. METHODS: Anti-allergic actions of artesunate were evaluated in passive cutaneous anaphylaxis and passive systemic anaphylaxis mouse models, and in ovalbumin-induced contraction of bronchial rings isolated from sensitized guinea pigs. Direct mast cell-stabilizing effect of artesunate was examined in RBL-2H3 mast cell line and in mature human cultured mast cells. Anti-allergic signaling mechanisms of action of artesunate in mast cells were also investigated. RESULTS: Artesunate prevented IgE-mediated cutaneous vascular hyperpermeability, hypothermia, elevation in plasma histamine level, and tracheal tissue mast cell degranulation in mice in a dose-dependent manner. In addition, artesunate suppressed ovalbumin-mediated guinea pig bronchial smooth muscle contraction. Furthermore, artesunate concentration-dependently blocked IgE-mediated degranulation of RBL-2H3 mast cells and human culture mast cells. Artesunate was found to inhibit IgE-induced Syk and PLCγ1 phosphorylation, production of IP(3) , and rise in cytosolic Ca(+2) level in mast cells. CONCLUSIONS: We report here for the first time that artesunate possesses anti-allergic activity by blocking IgE-induced mast cell degranulation, providing a foundation for developing artesunate for the treatment of allergic asthma and other mast cell-mediated allergic disorders.

Low-dose subcutaneous alemtuzumab is a safe and effective treatment for chronic acquired pure red cell aplasia.

Three patients with pure red cell aplasia, with or without co-existing large granular lymphocytic leukaemia, who remained transfusion-dependent despite treatment with established first-line therapy, were treated with low-dose subcutaneous alemtuzumab 15 mg twice to thrice per week, for 3 to 4 weeks. The mean response time was 17 days compared with a response time of at least 61 days on standard first-line therapy. There were no serious side-effects and the mean duration of remission was 13 months. Low-dose subcutaneous alemtuzumab is a safe and effective treatment for pure red cell aplasia and further trials should be conducted to compare the long-term effectiveness of this treatment with conventional therapy.

Localization of hRad9 in breast cancer.

BACKGROUND: hRad9 is a cell cycle checkpoint gene that is up-regulated in breast cancer. We have previously shown that the mRNA up-regulation correlated with tumor size and local recurrence. Immunohistochemical studies were made to better define the role of hRad9 in breast carcinogenesis. METHODS: Localisation of hRad9 protein were performed on paired tumor and normal breast tissues. Immunoblotting with and without dephosphorylation was used to define the protein isolated from breast cancer cells. RESULTS: Increased hRad9 protein was observed in breast cancer cells nucleus compared to non-tumor epithelium. This nuclear protein existed in hyperphosphorylated forms which may be those of the hRad9-hRad1-hHus1 complex. CONCLUSION: Finding of hyperphosphorylated forms of hRad9 in the nucleus of cancer cells is in keeping with its function in ameliorating DNA instability, whereby it inadvertently assists tumor growth.

The origin of ultra diffuse galaxies: stellar feedback and quenching.

We test if the cosmological zoom-in simulations of isolated galaxies from the FIRE project reproduce the properties of ultra diffuse galaxies (UDGs). We show that outflows that dynamically heat galactic stars, together with a passively aging stellar population after imposed quenching, naturally reproduce the observed population of red UDGs, without the need for high spin haloes, or dynamical influence from their host cluster. We reproduce the range of surface brightness, radius, and absolute magnitude of the observed red UDGs by quenching simulated galaxies at a range of different times. They represent a mostly uniform population of dark matter-dominated dwarf galaxies with M * ~ 108 M☉, low metallicity, and a broad range of ages; the more massive the UDGs, the older they are. The most massive red UDG in our sample(M * ~ 3 × 108 M☉) requires quenching at z ~ 3 when its halo reached M h ~ 1011M☉. Our simulated UDGs form with normal stellar-to-halo ratios and match the central enclosed masses and the velocity dispersions of the observed UDGs. Enclosed masses remain largely fixed across a broad range of quenching times because the central regions of their dark matter haloes complete their growth early. If our simulated dwarfs are not quenched, they evolve into bluer low surface brightness galaxies with M/L similar to observed field dwarfs. While our simulation sample covers a limited range of formation histories and halo masses, we predict that UDG is a common, and perhaps even dominant, galaxy type around M * ~ 108 M☉, both in the field and in clusters.

Gas kinematics in FIRE simulated galaxies compared to spatially unresolved HI observations.

The shape of a galaxy's spatially unresolved, globally integrated 21-cm emission line depends on its internal gas kinematics: galaxies with rotationally supported gas discs produce double-horned profiles with steep wings, while galaxies with dispersion-supported gas produce Gaussian-like profiles with sloped wings. Using mock observations of simulated galaxies from the FIRE project, we show that one can therefore constrain a galaxy's gas kinematics from its unresolved 21-cm line profile. In particular, we find that the kurtosis of the 21-cm line increases with decreasing V/σ and that this trend is robust across a wide range of masses, signal-to-noise ratios, and inclinations. We then quantify the shapes of 21-cm line profiles from a morphologically unbiased sample of ~2000 low-redshift, HI-detected galaxies with Mstar = 107-11 M☉ and compare to the simulated galaxies. At Mstar ≳ 1010 M☉, both the observed and simulated galaxies produce double-horned profiles with low kurtosis and steep wings, consistent with rotationally supported discs. Both the observed and simulated line profiles become more Gaussian like (higher kurtosis and less-steep wings) at lower masses, indicating increased dispersion support. However, the simulated galaxies transition from rotational to dispersion support more strongly: at Mstar 108-10 M, most of the simulations produce more Gaussian-like profiles than typical observed galaxies with similar mass, indicating that gas in the low-mass simulated galaxies is, on average, overly dispersion supported. Most of the lower-mass-simulated galaxies also have somewhat lower gas fractions than the median of the observed population. The simulations nevertheless reproduce the observed line-width baryonic Tully-Fisher relation, which is insensitive to rotational versus dispersion support.

Functional outcome of the hand following flexor tendon repair at the 'no man's land'.

PURPOSE: To evaluate the functional outcome of the hand following flexor tendon repair at 'no man's land' using 2 strands of a modified Kessler core suture and combined controlled motion rehabilitation protocol. METHODS: Records of 31 zone-2 flexor tendon injuries in 21 digits of 16 patients between July 2000 and June 2005 were reviewed retrospectively. The injured tendons were repaired within 24 hours using 2 strands of a modified Kessler core suture, reinforced by a continuous circumferential epitendon suture. All patients completed a rehabilitation protocol that included active extension against a rubber band, passive flexion, and controlled passive extension and passive flexion exercises. Functional outcome of the fingers was assessed using the Buck-Gramcko II score. Hand grip strength, rehabilitation period, and rupture rate were also measured. RESULTS: 17 (81%) out of 21 digits in 15 out of 16 patients achieved an excellent-to-good functional grade. The remaining patient with concomitant injuries to 4 (19%) digits attained a poor functional grade, attributable to poor compliance with the rehabilitation protocol. The mean rehabilitation period was 130 days and the mean grip strength was 78% that of the uninjured side. Concomitant digital nerve injury did not adversely affect the final outcome. Only one (4.8%) patient experienced a rupture. CONCLUSION: The surgical method and rehabilitation protocol used for zone-2 flexor tendon injury is safe and results in a reasonably good functional outcome.

Early clinical experience with a new preloaded one-piece intraocular lens in paediatric cataract surgery.

PurposeTo report the clinical experience of using the Tecnis PCB00 (Abbott Medical Optics, Santa Ana, CA, USA) preloaded one-piece intraocular lens (IOL) in the setting of a tertiary referral centre for paediatric cataract.MethodsA retrospective case note review of all paediatric cataract surgeries using the Tecnis PCB00 IOL, at a single UK paediatric ophthalmology department.ResultsNine eyes in seven patients received the IOL between December 2014 and January 2016. All patients underwent lens aspiration and insertion of the IOL 'in the bag.' The indications for surgery included developmental cataract (8/9) and traumatic cataract (1/9). Mean age at the time of surgery was 7 years (range 2-14). The median improvement in logMAR best-corrected visual acuity was 0.475 (range 0.250-1.500). The mean follow-up duration was 5 months (range 1-13). No operative or post-operative complications occurred as a result of using the device.ConclusionThe Tecnis PCB00 preloaded IOL appears to be a safe and effective device in treating paediatric cataract.

Peripheral T cell lymphoma.

Thirty-one Chinese patients with peripheral T cell lymphoma (PTCL) were reviewed. Using the modified Japanese Lymphoma Group classification, there were nine (29%) of the pleomorphic type, 16 (52%) immunoblastic lymphadenopathy (IBL)-like, two (7%) T-zone lymphoma, and one (3%) Lennert's lymphoepithelioid type. Three (9%) were not classifiable. All were positive for T11 (E rosette receptor antigen). Fifty-four percent (15 of 28) were positive predominantly for T4 (helper T cell) and 46% (13/28) for T8 (suppressor T cell). The median age of the patients was 57 years. They usually presented with advanced disease, and while extranodal involvement was common, CNS disease was not seen. The IBL-like type was associated with a positive Coombs' test and polyclonal hypergammaglobulinemia. Five of the nine pleomorphic type were checked for antibody to HTLV-I virus and all were negative. PTCL was associated with poor prognosis, which was not influenced by the histologic subtypes and the T4/T8 phenotypes. The complete response rate of 13 consecutive patients who received the BACOP (bleomycin, doxorubicin, cyclophosphamide, vincristine, and prednisone) L17M regimen was significantly better than the 16 historic controls who received other less-intensive regiments, 84% v 19% (P less than .01). The relapse rate was also significantly lower, 9% v 100% (P less than .001). There appeared to be an improvement in the disease-free survival (DFS) (80% v 0% at 18 months), as well as the overall survival (60% v 36% at 18 months), but the differences did not reach statistical significance due to small sample sizes.

Treatment outcome and prognostic factors for primary nasal lymphoma.

PURPOSE: To report our experience managing a large series of Chinese patients with primary nasal lymphoma. PATIENTS AND METHODS: From January 1975 to December 1993, 100 patients (median age, 50 years) with newly diagnosed primary nasal lymphoma were studied. There were four low-grade, 62 intermediate-grade, nine high-grade, and 25 unclassifiable lymphomas. Immunophenotyping was performed in 45 patients: eight B cell, 35 T cell, and two uncertain. All cases of angiocentric lymphoma that were typed were T cell. Fifty-two patients had stage I disease, 15 had stage II, four had stage III, and 29 had stage IV. Only 15 patients had B symptoms (weight loss, night sweats, and/or fever), and 11 had bulky disease. Thirty-nine patients with clinically localized stage I and II disease received local radiotherapy alone (before 1980), and the remaining 28 stage I and II patients received combination chemotherapy followed by local radiotherapy. The 33 patients with advanced stage III and IV disease were given combination chemotherapy, and additional radiotherapy was given to five of them who had bulky local disease. RESULTS: Significantly higher complete remission rates were observed in patients with early stages of disease and those without B symptoms. Superior disease-free survival after complete remission was observed in patients with stage I/II disease. Univariate factors associated with a better overall survival included age less than 60 years, stage I disease, and absence of B symptoms. Survival was significantly better in the subgroup of patients with stage I disease. CONCLUSION: Patients with nasal lymphoma, especially those with advanced disease, seemed to have a poor prognosis, and their clinical outcome was not improved significantly by the use of chemotherapy instead of radiotherapy or the use of doxorubicin-containing chemotherapeutic regimens.

Combined use of warfarin and adjusted subcutaneous heparin during pregnancy in patients with an artificial heart valve.

Adjusted subcutaneous heparin was used for thromboembolism prophylaxis during 18 pregnancies in 16 women with an artificial heart valve. Oral warfarin was replaced by subcutaneous heparin as soon as pregnancy was confirmed. The dosage of heparin was adjusted to maintain a partial thromboplastin time at 1.5 times the control value and treatment was administered during the first trimester and the last 3 weeks of gestation. Warfarin was used between the 13th and 37th week. There were no maternal thromboembolic complications and none of the live-born infants showed congenital malformations, indicating that this regimen is effective. However, there were nine spontaneous abortions, including five that occurred in the first 12 weeks. The early abortions were probably related to warfarin exposure at the beginning of pregnancy. The preconception replacement of warfarin by heparin in these patients may be indicated.

Detection of T-cell receptor delta gene rearrangement by clonal specific polymerase chain reaction.

A sensitive and specific technique to detect minimal residual disease for T-cell malignancies was explored. Southern analysis and polymerase chain reaction (PCR) were used to detect the rearranged V-D-J segment of T-cell receptor delta (TCR delta) gene from malignant cell specimens of patients with leukemia and lymphoma of T-cell lineage. The PCR product was sequenced and from the DNA sequences of the V-D-J region, a 3' antisense primer was designed and synthesized for clonal specific PCR (CS-PCR). Seven of the 22 T-ALL (32%) and 5 of 18 (28%) T-cell lymphoma showed clonal rearrangement by Southern analysis. Six of the 7 (86%) T-ALL and 4 of the 5 (80%) T-cell lymphoma which were Southern positive were also positive by TCR delta PCR. The PCR products of four cases of T-ALL showing clonal pattern by TCR delta PCR amplification were successfully sequenced and CS-PCR amplification performed. CS-PCR detected with confidence specific clonal rearrangement in a mixture containing 0.003% of malignant cells. Marrow specimens obtained at diagnosis and subsequent follow-ups from the 4 T-ALL patients were studied by Southern analysis, TCR delta PCR and CS-PCR. The first patient was in continuous morphological complete remission for more than 3 years and had persistently negative Southern, TCR delta PCR and CS-PCR results on follow-up. Initial follow-up marrow samples from the second patient had persistently positive CS-PCR results while they were still morphologically and TCR delta PCR negative and the patient had a frank leukemic relapse at 18 months. The other two patients had persistent disease by conventional morphological examination, Southern analysis, TCR delta PCR and CS-PCR studies were all positive as expected. CS-PCR is a highly specific and sensitive technique in detecting minimal residual disease for T-cell malignancies. Its potential applications warrant further clinical evaluation and correlation.

Poor outcome of intensive chemotherapy for adult acute lymphoblastic leukemia: a possible dose effect.

Fifty consecutive adult patients with acute lymphoblastic leukemia (ALL) were treated with an intensive cyclical chemotherapy and the mean received dose of individual cytotoxic drug was retrospectively studied. The median age was 28 years. Twenty-one (43%) had white blood cell (WBC) count over 30 x 10(9)/l. Of the 26 patients with successful cytogenetic studies, ten (28%) had unfavorable clonal chromosomal abnormalities (four Philadelphia chromosome, six others). A high complete remission (CR) rate (86%) was achieved. This was associated with delivery of 100% of the planned dosage of vincristine, prednisone, and daunorubicin at induction. Dose reduction of asparaginase, the fourth drug in the induction protocol, was recorded in 20 (40%) patients. The CR rate of these patients was not adversely affected. Dose reduction was recorded during consolidation (38 of 43 remitters) and maintenance (18 of 20 remitters) as a result of treatment toxicity. The mean received dose of teniposide, Ara-C, asparaginase, mercaptopurine, and methotrexate was 73% (SD 7%), 73% (SD 7%), 62% (SD 41%), 65% (SD 15%) and 73% (SD 17%) of the planned dosage, respectively. The 5-year overall survival and leukemia-free survival (LFS) were 11% (95% CI: 0-27%) and 13% (95% CI: 0-26%), respectively. Even standard-risk patients had 4-year LFS of only 26% (95% CI: 0-57%). Among 36 remitters not withdrawn from consolidation, there were 29 treatment failures after a median follow-up of 42 months; 25 (86%) of these were leukemia relapse, three (10%) were toxic death during consolidation, and one patient (4%) died from therapy-related myelodysplastic syndrome. We postulate inadequate drug delivery during postremission therapy contributed to the high relapse rate in the whole group as well as the standard-risk patients.

Acute intravascular hemolysis indicating thrombosis of Bjork-Shiley aortic prosthesis.

A patient with a Bjork-Shiley aortic prosthesis inserted two years and four months prior to receiving adequate anticoagulant therapy suddenly had severe intravascular hemolysis develop as a result of thrombosis of her aortic prosthesis. There was no notable hemodynamic compromise. The mechanism of the intravascular hemolysis is discussed.

The effect of triamterene and sodium intake on renin, aldosterone, and erythrocyte sodium transport in Liddle's syndrome.

Liddle's syndrome was diagnosed in a 23-yr-old Chinese girl with hypertension and hypokalemia by the presence of suppressed renin and negligible plasma and urinary aldosterone secretion. Adrenal corticosteroids, including aldosterone, were suppressed by dexamethasone and stimulated by ACTH. While spironolactone was ineffective, triamterene (2,4,7-triamino-6-phenyl-pteridine) treatment corrected the hypertension and hypokalemia and restored PRA to normal provided that sodium intake was not excessive. During long term treatment with triamterene, blood pressure was extremely sensitive to salt intake, increasing promptly with high intake and decreasing with low salt intake. As a result of the chronic hypervolemia and sodium retention consequent upon the patient's persistent high salt intake and increased renal tubular sodium reabsorption, plasma renin and aldosterone remained low. Erythrocyte sodium concentration and membrane permeability were increased. Triamterene with salt restriction was able to lower the intracellular sodium concentration but did not correct the increased sodium permeability. This suggests that there is an abnormality of sodium transport in Liddle's syndrome which affects the erythrocytes as well as the renal tubular cells.

A unique patient with coexisting cerebrotendinous xanthomatosis and beta-sitosterolemia.

An adult Chinese man presented with tendinous and tuberous xanthomatosis and severe atheromatous changes in the coronary arteries. In addition, he had chronic hemolytic anemia, with spherostomatocytic erythrocytes. Cerebrotendinous xanthomatosis was diagnosed on the basis of increased cholestanol levels in his plasma, red cells and xanthoma, changes in bile acid composition due to the defective synthesis of chenodeoxycholic acid. Coexisting beta-sitosterolemia was confirmed by the finding of large amounts of the plant sterols such as beta-sitosterol and campesterol. This is the first report of these two rare lipid storage disorders in the same patient.

Cell-free synthesis of factor VIII related protein.

Polyadenylated ribonucleic acid (RNA) was isolated from confluent human endothelial cells in culture and translated in a cell-free rabbit reticulocyte lysate protein-synthesizing system. Using specific immunoprecipitation, the translation product was found to contain 1.58% VIII related protein (VIII R), which has a molecular weight of 0.26 X 10(6), similar to the subunit of VIII R present in normal plasma. This indicated that VIII R messenger RNA is responsible for the synthesis of subunits only and intracellular post-translational events are required for the assembly of tetramer and multimers.

Characterization of factor VIII related protein synthesized by human endothelial cell: a study of structure and function.

Crossed immunoelectrophoresis showed that factor VIII related protein (VIII R) synthesized in the human endothelial cell (EC-VIII R) had faster electrophoretic mobility than that secreted into the culture medium (MED-VIII R). Sodium dodecyl sulphate agarose gel electrophoresis followed by radioimmunofixation showed that EC-VIII R consisted of molecules varying from 0.26 x 10(6) to 3.76 x 10(6) daltons while MED-VIII R had molecules ranging from 0.93 x 10(6) to greater than 10 x (10)6 daltons, similar to that present in plasma. The smallest VIII R molecule present in normal plasma or spent culture medium (0.93 x 10(6) daltons) corresponded to a tetramer of subunits of 0.22-0.24 x 10(6) daltons. Only molecular forms greater than 3.76 x 10(6) daltons possessed ristocetin cofactor activity. Sonication (15 mu amplitude for 30 secs) effectively broke the non-covalent bonds of the VIII R multimers resulting in smaller molecules. Thus endothelial cells in culture synthesized VIII R subunits and assembled them into the higher multimeric forms on secretion. Different types of von Willebrand disease could result from defects of either of the two processes.

Antithrombin III, the major modulator of intravascular coagulation, is synthesized by human endothelial cells.

Human endothelial cells in culture is shown to synthesize antithrombin III (At-III). The endothelial cell At-III (EC-At-III) consists of a small fraction similar to plasma At-III and a larger fraction with decreased heparin-binding as tested by crossed immunoelectrophoresis. However, both the anti-Xa and thrombin-neutralizing activities of the EC-At-III were rapid and active even in the absence of added heparin. It is concluded that the major portion was probably bound to endogenous heparin-like substance, thus accounting for its decreased exogenous heparin binding. The presence of At-III and other antithrombotic factors in the vascular endothelium offer protection against thrombosis and possibly atherosclerosis.