Ultrasound-mediated gene transfection in vitro: effect of ultrasonic parameters on efficiency and cell viability.

Ultrasound (US)-mediated gene transfection in the presence of microbubbles is a recently developed and promising non-viral gene delivery method. Optimising the parameters used in ultrasonic transfection is urgently required in order to realise higher transfection efficiencies in clinical settings. This study examined the effect of ultrasound exposure parameters on plasmid DNA transfection in mouse embryonic fibroblast cell lines using perfluorobutane bubbles. Variations in US intensity (0-11 W/cm2), pulse repetition frequency (PRF, 50-50,000 Hz), duty ratio (10 to 50%), exposure time (0-120 s) and microbubble volume concentration (0 to 10%) were tested, and the microbubble volume concentration was also monitored during exposure. Through the experiments, the mechanism of how variations in parameters influence US-mediated gene transfection was discussed, which can provide a basis for future applications of ultrasound mediated transfection.

A case of congenital dyserythropoietic anemia type 1 in a Japanese adult with a CDAN1 gene mutation and an inappropriately low serum hepcidin-25 level.

We describe the first case of genetically diagnosed congenital dyserythropoietic anemia (CDA) type 1 in a Japanese man. The patient had hemolytic anemia since he was a child, and he developed diabetes, hypogonadism, and liver dysfunction in his thirties, presumably from systemic iron overload. When he was 48 years old a diagnosis was finally made by genetic analysis that revealed a homozygous mutation of CDAN1 gene (Pro1129Leu). His serum hepcidin-25 level was inappropriately low. We conclude that physicians should be aware of the possibility of CDA in a patient with anemia and systemic iron overload at any age.

The role of interleukin-6 in a patient with polyclonal hairy B-cell lymphoproliferative disorder: a case report.

An 80-year-old female patient showed persistent lymphocytosis morphologically resembling the Japanese variant of hairy cell leukemia (HCL). However, flow cytometric analysis determined that these lymphocytes were of polyclonal B-cell origin, showing CD5-, CD10(-), CD11c(+), CD19(+), CD20(+), CD23(-), CD103(-), FMC7(-), HLA-DR(+) and surface membrane immunoglobulin (smIg) G(+) phenotype. The female patient also showed polyclonal hypergammaglobulinemia with bone marrow plasmacytosis. The patient was diagnosed as having hairy B-cell lymphoproliferative disorder (HBLD). Serum interleukin-6 (IL-6) level was elevated at the time of diagnosis in this patient, but IL-6 receptor (CD126) was not expressed on the hairy B-cells. Intracellular IL-6 was not detected in these cells either, suggesting that IL-6 did not play an important role in the B-lymphocytosis present in our patient with HBLD.

[Improvement of thrombocytopenia with normalization of thyroid function in a patient with Graves disease].

A 39-year-old man was admitted to our hospital because of hyperthyroidism complicated with fever, atrial fibrillation with rapid ventricular response and congestive heart failure. Laboratory data revealed pancytopenia with a white blood cell count of 3360/microl, Hb 9.6 g/dl, and a platelet count of 88000/ul. After initiation of treatment with 30 mg of thiamazole daily, the patient's thyroid function improved but 4 weeks later, thiamazole induced agranulocytosis occurred and the drug was discontinued. After recovery from agranulocytosis, a subtotal thyroidectomy was performed, and the patient's pancytopenia rapidly improved. In this case, hyperthyroidism itself appeared to be closely related to the development of thrombocytopenia.

Cytotoxic T-cell non-Hodgkin's lymphoma of the thyroid gland.

An 86-year-old female was diagnosed with peripheral T-cell lymphoma, unspecified, which affected only her thyroid gland. Lymphoma cells were TIA-1(+), suggesting cytotoxic T-cell origin. She had not suffered from autoimmune thyroiditis and showed normal thyroid function. Without any specific therapies, the lymphomatous lesion showed the partial spontaneous regression before diagnostic hemithyroidectomy, and she had been well for 2 years without relapse. This is the first case of primary thyroid lymphoma of cytotoxic T-cell origin that showed spontaneous regression.

Quinine-resistant severe falciparum malaria effectively treated with atovaquone and proguanil hydrochloride combination therapy.

A 22-year-old Japanese man noticed pyrexia and diarrhea after travel to Guinea. Notable physical findings included hepatosplenomegaly. Treatment with oral quinine and minocycline was started after definitive diagnosis of falciparum malaria by blood smear. Initially, parasitemia and body temperature decreased but by the third night of therapy his temperature increased to 40 degrees C with a slight increase of parasite count. When quinine treatment was changed to atovaquone/proguanil, his temperature dropped immediately and complete plasmodial elimination was confirmed on microscopic examination. Subsequent recrudescence of the disease was not observed. It was concluded that the antimalarial treatment with atovaquone/proguanil might become invaluable in Japan.

Electro-spinning and electro-blowing of hyaluronic acid.

In this study, hyaluronic acid (HA) was electro-spun and electro-blown to prepare nonwoven nanofibrous membranes. Critical parameters for processing and corresponding effects on the membrane morphology were investigated using the methods of rheology and scanning electron microscopy (SEM). During electro-spinning, the optimal HA concentration window for nanofibrous formation was determined within a narrow range of 1.3-1.5 w/v %, corresponding to a solution viscosity range of 3-30 Pa s at a shear rate of 1 s(-1). SEM results indicated that, with increases in (1) the total concentration by blending of low molecular weight HA, (2) the evaporation rate by the addition of ethanol, and/or (3) the feeding rate of solution, the electro-spinning performance for creating nanofibers was improved. However, the improvement was not sufficient to achieve a consistent production of high quality nonwoven nanofiber membranes. This problem was overcome by a new electro-blowing process using the combination of air flow and electro-spinning. Although air blowing at room temperature around the spinneret orifice did not exhibit a remarkable enhancement of nanofiber formation of HA, the performance was significantly improved with an increase in the air blowing rate. SEM results showed that the temperature of air-blowing was the most effective parameter in ensuring HA nanofiber formation. As the temperature of the blown air increased from 25 to 57 degrees C, the nanofiber formation became consistent and uniform. A high quality HA nonwoven membrane of nanofibers was successfully produced by blowing air at 57 degrees C with a 70 ft(3)/hr flow rate.

Asymmetrical-flow field-flow fractionation with on-line multiangle light scattering detection. 1. Application to wormlike chain analysis of weakly stiff polymer chains.

Four samples of hyaluronan in the sodium form, ranging in weight-average molecular weight, M(w), from 6.67 x 10(5) to 4.23 x 10(6) were investigated by asymmetrical-flow field-flow fractionation coupled to multiangle light scattering (FlFFF-MALS) in 0.2 M aqueous NaCl at 25 degrees C. M(w) and z-average radii of gyration, R(G)(z)(), obtained via FlFFF-MALS showed a good agreement with the results obtained by conventional static light scattering. Furthermore, the molecular weight dependence of the radius of gyration for sodium hyaluronan obtained via FlFFF-MALS was analyzed on the basis of the Kratky-Porod model for unperturbed wormlike chains combined with the Yamakawa theory for radius expansion factor, and a sufficiently good agreement was observed between the theoretical prediction and experimental data. These results show the potential usage of FlFFF-MALS regarding size separation and molecular characterization even for weakly stiff chains.

Infusion of a high number of CD34+ cells provides a rapid hematopoietic recovery and cost savings in autologous peripheral blood stem cell transplantation.

BACKGROUND: The objectives of this study were to evaluate the effect of the number of infused CD34+ cells on hematopoietic recovery and on the cost in autologous peripheral blood stem cell transplantation (PBSCT). METHODS: Sixty-nine patients who received autologous PBSCT (ABSCT) were divided into three groups defined by the number of infused CD34+ cells. The number of days until 0.5 x 10(9)/l neutrophils and 50 x 10(9)/l platelets, the number of transfused blood products, the febrile days, the duration of parenteral antibiotics and the cost of additional supportive care (transfusions of blood products and parenteral antibiotics) were analyzed. RESULTS: Twenty-three patients received <2.5 x 10(6)/kg of CD34+ cells (group A), 25 patients received > or = 2.5 to 5 x 10(6)/kg of CD34+ cells (group B) and 21 patients received > or = 5 x 10(6)/kg of CD34+ cells (group C). Patients in group C had rapid neutrophil (p < 0.01) and platelet (p < 0.05) recovery and required less platelet transfusions (p < 0.05) than patients in other groups. Transfusions of red blood cell concentrates, the duration of febrile days or parenteral antibiotics were not statistically different between the two groups. The patients in group C required significantly lower costs for platelet concentrates and additional supportive care (p < 0.05). CONCLUSION: Infusion of > or = 5 x 10(6)/kg of CD34+ cells in ABSCT shortens hematopoietic recovery and reduces costs for additional supportive care.