PTD-Mediated Red Blood Cell Encapsulation of L-Asparaginase for Potential Treatment of Acute Lymphoblastic Leukemia.

One of the primary drugs used in treatment of acute lymphoblastic leukemia is L-asparaginase (ASNase), which despite its therapeutic efficacy, faces challenges often faced in protein drug treatment: the immunogenicity and short half-life of the drug. In order to overcome these major challenges, the red blood cell (RBC)encapsulation method has been employed to prevent degradation by serum proteases and elimination by reticuloendothelial system, thereby increasing circulation half-life of the protein drug. However, the encapsulation methods used thus far, such as hypotonic dialysis/resealing and electroporation, showed compromise in the oxygen transport function and structural integrity of RBCs. Herein we introduce a novel RBC encapsulation method that involves low molecular weight protamine (LMWP), a protein transduction domain (PTD) peptide. Protein drug was first conjugated to LMWP via disulfide bond and the cell penetrating property of LMWP allowed encapsulation of protein drug into RBCs as verified by the confocal microscopy images. The optimal encapsulation condition was determined to be 37 °C for 30 minutes. Hematological parameters as well as SEM and osmotic fragility curve of LMWP-ASNase encapsulated RBCs showed that the structural integrity was maintained while Hill coefficients and pO50 values indicated oxygen transport functionality was maintained. LMWP-mediated encapsulation method considerably increased the circulating half-life of ASNase compared to that of the hypo-osmotic rupture/resealing method and improved the survival time of tumor-bearing mice by 44% compared to the saline control group. Based on these data, novel method for encapsulating protein drugs into intact and fully functional RBCs was established for potential treatment of acute lymphoblastic leukemia.Ph.D.Pharmaceutical SciencesUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/75888/1/heec_1.pd

Eosinophilic myositis in a slaughtered Korean native cattle

Histopathological findings of eosinophilic myositis in the carcass of a slaughtered Korean native cow are presented. Lesions contained massive fibrous septae with vacuolar changes in some lesions, and the hypercontraction and rupturing of muscle bundles, with replacement by eosinophils. Necrosis and severe eosinophil infiltration were observed. Sarcoplasmic fragmentation and atrophy developed. Typical of granuloma, calcified myofibers were focally surrounded by macrophages and numerous inflammatory cells, and multinucleated giant cell formation was evident

Properties of Central Caustics in Planetary Microlensing

To maximize the number of planet detections, current microlensing follow-up observations are focusing on high-magnification events which have a higher chance of being perturbed by central caustics. In this paper, we investigate the properties of central caustics and the perturbations induced by them. We derive analytic expressions of the location, size, and shape of the central caustic as a function of the star-planet separation, $s$, and the planet/star mass ratio, $q$, under the planetary perturbative approximation and compare the results with those based on numerical computations. While it has been known that the size of the planetary caustic is \propto \sqrt{q}, we find from this work that the dependence of the size of the central caustic on $q$ is linear, i.e., \propto q, implying that the central caustic shrinks much more rapidly with the decrease of $q$ compared to the planetary caustic. The central-caustic size depends also on the star-planet separation. If the size of the caustic is defined as the separation between the two cusps on the star-planet axis (horizontal width), we find that the dependence of the central-caustic size on the separation is \propto (s+1/s). While the size of the central caustic depends both on $s$ and q, its shape defined as the vertical/horizontal width ratio, R_c, is solely dependent on the planetary separation and we derive an analytic relation between R_c and s. Due to the smaller size of the central caustic combined with much more rapid decrease of its size with the decrease of q, the effect of finite source size on the perturbation induced by the central caustic is much more severe than the effect on the perturbation induced by the planetary caustic. Abridged.Comment: 5 pages, 4 figures, ApJ accepte

Hearing Abilities at Ultra-High Frequency in Patients with Tinnitus

ObjectivesTo compare tinnitus patients who have normal hearing between 250 Hz and 8 kHz with normal controls with regard to the ability of each group to hear extended high-frequency pure tone thresholds.MethodsWe enrolled 18 tinnitus patients, each of whom had a threshold of HL <25 dB and threshold differences of <10 dB between ears at frequencies of 250 and 500 Hz and 1, 2, 4, and 8 kHz. We also enrolled age- and gender-matched normal volunteers (10 ears), for each patient. Extended high frequency pure tone audiometry was performed, and the mean hearing thresholds at 10, 12, 14, and 16 kHz of each tinnitus ear were compared with those of the 10 age- and sex-matched normal ears.ResultsOf the 18 patients with tinnitus, 12 had significantly increased hearing thresholds at more than one of the four high frequencies, compared with the normal group. When we assessed results according to frequency, we found that 8 patients had decreased hearing ability at 10 kHz, 10 at 12 kHz, 8 at 14 kHz, and 4 at 16 kHz.ConclusionSome patients with tinnitus who have normal hearing below 8 kHz have decreased hearing ability at extended high-frequencies. Thus, the proportion of patients with tinnitus who have normal hearing over the entire audible range is smaller than in previous reports

Genome-wide genetic aberrations of thymoma using cDNA microarray based comparative genomic hybridization

BACKGROUND: Thymoma is a heterogeneous group of tumors in biology and clinical behavior. Even though thymoma is divided into five subgroups following the World Health Organization classification, the nature of the disease is mixed within the subgroups. RESULTS: We investigated the molecular characteristics of genetic changes variation of thymoma using cDNA microarray based-comparative genomic hybridization (CGH) with a 17 K cDNA microarray in an indirect, sex-matched design. Genomic DNA from the paraffin embedded 39 thymoma tissues (A 6, AB 11, B1 7, B2 7, B3 8) labeled with Cy-3 was co-hybridized with the reference placenta gDNA labeled with Cy-5. Using the CAMVS software, we investigated the deletions on chromosomes 1, 2, 3, 4, 5, 6, 8, 12, 13 and 18 throughout the thymoma. Then, we evaluated the genetic variations of thymoma based on the subgroups and the clinical behavior. First, the 36 significant genes differentiating five subgroups were selected by Significance Analysis of Microarray. Based on these genes, type AB was suggested to be heterogeneous at the molecular level as well as histologically. Next, we observed that the thymoma was divided into A, B (1, 2) and B3 subgroups with 33 significant genes. In addition, we selected 70 genes differentiating types A and B3, which differ largely in clinical behaviors. Finally, the 11 heterogeneous AB subtypes were able to correctly assign into A and B (1, 2) types based on their genetic characteristics. CONCLUSION: In our study, we observed the genome-wide chromosomal aberrations of thymoma and identified significant gene sets with genetic variations related to thymoma subgroups, which might provide useful information for thymoma pathobiology.ope

Ductographic Findings of Breast Cancer

Ductography has become the gold standard for the evaluation of patients exhibiting pathologic nipple discharges. In nine patients (age range, 29-67 years; median age, 51 years) with invasive (n=5) or intraductal (n=4) cancer, ductographic findings were recorded, then correlated with mammographic and sonographic findings. Common ductographic findings included complete ductal obstruction, multiple irregular filling defects in the nondilated peripheral ducts, ductal wall irregularities, periductal contrast extravasation, and ductal displacement. Faint microcalcifications or ill-defined masses, which were not opacified by contrast material, were often discovered adjacent to ductal abnormalities. Mammographically and sonographically occult diffusely spreading intraductal cancers often manifested as pathologic nipple discharge. In such cases, meticulous ductographic examinations and interpretations were crucial in order not to miss breast cancers

PI3K-Akt-Wnt Pathway Is Implicated in Exercise-Induced Improvement of Short-term Memory in Cerebral Palsy Rats

Purpose Maternal lipopolysaccharide (LPS) injection induces neurodevelopmental disorders, such as cerebral palsy. Exercise activates phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signaling pathway that enhances neurogenesis. Wnt ligands are also implicated in the hippocampal neurogenesis and synaptic plasticity. Glycogen synthase kinase-3β (GSK-3β) is a downstream molecule of Akt, and GSK-3β is known to modulate hippocampal neurogenesis negatively. Methods Cerebral palsy was made by maternal LPS-injection. On the 5 weeks after birth, treadmill running was applied to the rat pups of the exercise groups, for 30 minutes, 5 times a week during 6 weeks. Results Treadmill running alleviated short-term memory impairments of the cerebral palsy rat pups. Hippocampal cell proliferation was increased and hippocampal apoptosis was suppressed by treadmill running in the cerebral palsy rat pups. Hippocampal phosphorylated-PI3K/PI3K ratio, phosphorylated-Akt/Akt ratio, and Wnt expression were enhanced by treadmill running in the cerebral palsy rat pups. In contrast, hippocampal phosphorylated-GSK-3β/GSK-3β ratio and β-catenin expression were suppressed by treadmill running in the cerebral palsy rat pups. Conclusions The results of this study showed that short-term memory improvement due to treadmill running in cerebral palsy occurs via activation of the PI3K-Akt-Wnt pathway

Rubus Crataegifolius Bunge Regulates Adipogenesis Through Akt and Inhibits High-Fat Diet-Induced Obesity in Rats

BACKGROUND: Obesity is one of the greatest public health problems and major risk factors for serious metabolic diseases and significantly increases the risk of premature death. The aim of this study was to determine the inhibitory effects of Rubus crataegifolius Bunge (RCB) on adipocyte differentiation in 3 T3-L1 cells and its anti-obesity properties in high fat diet (HFD)-induced obese rats. METHODS: 3 T3-L1 adipocytes and HFD-induced obese rats were treated with RCB, and its effect on gene expression was analyzed using RT-PCR and Western blotting experiments. RESULTS: RCB treatment significantly inhibited adipocyte differentiation by suppressing the expression of C/EBPβ, C/EBPα, and PPARγ in the 3 T3-L1 adipocytes. Subsequently, the expression of the PPARγ target genes aP2 and fatty acid synthase (FAS) decreased following RCB treatment during adipocyte differentiation. In uncovering the specific mechanism that mediates the effects of RCB, we demonstrated that the insulin-stimulated phosphorylation of Akt strongly decreased and that its downstream substrate phospho-GSK3β was downregulated following RCB treatment in the 3 T3-L1 adipocytes. Moreover, LY294002, an inhibitor of Akt phosphorylation, exerted stronger inhibitory effects on RCB-mediated suppression of adipocyte differentiation, leading to the inhibition of adipocyte differentiation through the downregulation of Akt signaling. An HFD-induced obesity rat model was used to determine the inhibitory effects of RCB on obesity. Body weight gain and fat accumulation in adipose tissue were significantly reduced by the supplementation of RCB. Moreover, RCB treatment caused a significant decrease in adipocyte size, associated with a decrease in epididymal fat weight. The serum total cholesterol (TC) and triglyceride (TG) levels decreased in response to RCB treatment, whereas HDL cholesterol (HDL-C) increased, indicating that RCB attenuated lipid accumulation in adipose tissue in HFD-induced obese rats. CONCLUSION: Our results demonstrate an inhibitory effect of RCB on adipogenesis through the reduction of the adipogenic factors PPARγ, C/EBPα, and phospho-Akt. RCB had a potent anti-obesity effect, reducing body weight gain in HFD-induced obese rats