Nanohydroxyapatite/Peptide Composite Coatings on Pure Titanium Surfaces with Nanonetwork Structures Using Oyster Shells.

Titanium and its alloys are extensively applied in artificial tooth roots because of their excellent corrosion resistance, high specific strength, and low elastic modulus. However, because of their biological inertness, their surface needs to be modified to improve the osteointegration of titanium implants. The preparation of biologically active calcium-phosphorus coatings on the surface of an implant is one effective method for enhancing the likelihood of bone integration. In this study, osteoinductive peptides were extracted from oyster shells by using acetic acid. Two peptide-containing hydroxyapatite (HA) composite coatings were then prepared: one coating was prepared by hydrothermally synthesizing an HA coating in the presence of peptides (HA/P/M), and the other coating was prepared by hydrothermally synthesizing HA and then immersing the hydrothermally synthesized HA in a peptide solution (HA/P/S). Characterization results indicated that the composite HA coatings containing oyster shell-based peptides were successfully prepared on the alkali-treated pure titanium surfaces. The HA/P/M and HA/P/S composite coatings were found to exhibit excellent hydrophilicity. Protein adsorption tests confirmed that the HA/P/M and HA/P/S coatings had an approximately 2.3 times higher concentration of adsorbed proteins than the pure HA coating.

Actin filaments form a size-dependent diffusion barrier around centrosomes.

The centrosome, a non-membranous organelle, constrains various soluble molecules locally to execute its functions. As the centrosome is surrounded by various dense components, we hypothesized that it may be bordered by a putative diffusion barrier. After quantitatively measuring the trapping kinetics of soluble proteins of varying size at centrosomes by a chemically inducible diffusion trapping assay, we find that centrosomes are highly accessible to soluble molecules with a Stokes radius of less than 5.8 nm, whereas larger molecules rarely reach centrosomes, indicating the existence of a size-dependent diffusion barrier at centrosomes. The permeability of this barrier is tightly regulated by branched actin filaments outside of centrosomes and it decreases during anaphase when branched actin temporally increases. The actin-based diffusion barrier gates microtubule nucleation by interfering with γ-tubulin ring complex recruitment. We propose that actin filaments spatiotemporally constrain protein complexes at centrosomes in a size-dependent manner.

Inactivation of Escherichia coli by dual-functional zerovalent Fe/Al composites in water.

A bimetallic Fe/Al disinfection system was developed to examine the feasibility of inactivation of water borne microorganisms. In this study, the effectiveness and mechanisms of the bimetallic Fe/Al system on the inactivation of model bacteria, Escherichia coli (E. coli), were investigated. Results revealed that the Fe/Al system effectively inactivated E. coli to reach nearly 2 logs (99%) removal within 20 min and 4 logs (99.99%) at 24 h, indicating that the Fe/Al composite was able to sustain a long-term disinfection capacity. The inactivation ability resulted from hydroxyl radicals produced by a Fenton reaction through in-situ self-generated Fe2+ and H2O2 species in the Fe/Al system. In addition to the attack by the radicals, some of E. coli were adsorbed onto the Fe/Al composite (zeta potential of 30-50 mV) as a result of Coulomb interaction. Scanning electron microscope (SEM) images showed that the adsorbed bacteria had damaged pores at the two ends of their rod-like cells. This phenomenon suggested that a micro-electric field between the Fe/Al galvanic couple induced electroporation of the adsorbed E. coli and thus further advanced additional inactivation ability for the bacteria disinfection.

Favorable mortality-to-incidence ratios of kidney Cancer are associated with advanced health care systems.

BACKGROUND: The advancements in cancer therapy have improved the clinical outcomes of cancer patients in recent decades. However, advanced cancer therapy is expensive and requires good health care systems. For kidney cancer, no studies have yet established an association between clinical outcome and health care disparities. METHODS: We used the mortality-to-incidence ratio (MIR) for kidney cancer as a marker of clinical outcome to compare World Health Organization (WHO) country rankings and total expenditures on health/gross domestic product (e/GDP) using linear regression analyses. RESULTS: We included 57 countries based on data from the GLOBOCAN 2012 database. We found that more highly developed regions have higher crude and age-standardized rates of kidney cancer incidence and mortality, but a lower MIR, when compared to less developed regions. North America has the highest crude rates of incidence, but the lowest MIRs, whereas Africa has the highest MIRs. Furthermore, favorable MIRs are correlated with countries with good WHO rankings and high e/GDP expenditures (p < 0.001 and p = 0.013, respectively). CONCLUSIONS: Kidney cancer MIRs are positively associated with the ranking of health care systems and health care expenditures.

Manipulating Cellular Activities Using an Ultrasound-Chemical Hybrid Tool.

We developed an ultrasound-chemical hybrid tool to precisely manipulate cellular activities. A focused ultrasound coupled with gas-filled microbubbles was used to rapidly trigger the influx of membrane-impermeable chemical dimerizers into living cells to regulate protein dimerization and location without inducing noticeable toxicity. With this system, we demonstrated the successful modulation of phospholipid metabolism triggered by a short pulse of ultrasound exposure. Our technique offers a powerful and versatile tool for using ultrasound to spatiotemporally manipulate the cellular physiology in living cells.

The gender difference and mortality-to-incidence ratio relate to health care disparities in bladder cancer: National estimates from 33 countries.

The variation in the mortality-to-incidence ratio (MIR) between countries and genders reflects the complex etiology and intervention of bladder cancer. In this study, we investigated the MIR variation between genders and health care disparities among countries. Cancer incidence and mortality were obtained from the GLOBOCAN 2012 database. The ranking and the total expenditure on health of countries were obtained from the World Health Organization. Linear regression was used to estimate the significance between variables. We estimated the role of MIRs from 33 countries. Bladder cancer incidence and mortality rates were higher in more developed regions, Europe, and the Americas. The MIRs were higher in less developed regions. Analysis according to country revealed Germany to have the lowest MIR. High relative MIRs (female MIR/male MIR) for bladder cancer were noted in many developed countries. A correlation between MIR and health care disparities among countries was indicated by a significant association between the World Health Organization ranking and total expenditure on health/GDP with the MIR and relative MIR. Low bladder cancer MIR is prone to be more prevalent in countries with good health care system.

Prostate Cancer Mortality-To-Incidence Ratios Are Associated with Cancer Care Disparities in 35 Countries.

The variation in mortality-to-incidence ratios (MIRs) among countries reflects the clinical outcomes and the available interventions for colorectal cancer treatments. The association between MIR of prostate cancer and cancer care disparities among countries is an interesting issue that is rarely investigated. For the present study, cancer incidence and mortality rates were obtained from the GLOBOCAN 2012 database. The rankings and total expenditures on health of various countries were obtained from the World Health Organization (WHO). The association between variables was analyzed by linear regression analyses. In this study, we estimated the role of MIRs from 35 countries that had a prostate cancer incidence greater than 5,000 cases per year. As expected, high prostate cancer incidence and mortality rates were observed in more developed regions, such as Europe and the Americas. However, the MIRs were 2.5 times higher in the less developed regions. Regarding the association between MIR and cancer care disparities, countries with good WHO ranking and high total expenditures on health/gross domestic product (GDP) were significant correlated with low MIR. The MIR variation for prostate cancer correlates with cancer care disparities among countries further support the role of cancer care disparities in clinical outcome.

Prostate Lymphoma Complicated With Prostatorectal Fistula and Multifocal Infection After Chemotherapy.

A 67-year-old man with a biopsy proving prostate diffuse large B-cell lymphoma was referred for an F-FDG PET/CT scan for postchemotherapy treatment monitoring. The FDG PET/CT scan revealed a significant reduction of prostate lymphoma and a prostatorectal fistula formation. The patient developed Klebsiella pneumoniae sepsis, and a follow-up FDG PET/CT 3 months later revealed multifocal lung, bone, and muscle infection. The FDG PET/CT scans of this patient not only demonstrated the effectiveness of chemotherapy for prostate lymphoma but also the complications of a prostatorectal fistula and multiple infectious foci.

Dynamic FDG PET/CT imaging with diuresis demonstrates an enterovesical fistula in a lymphoma patient with repeated colon diverticulitis.

A 43-year-old male patient with follicular B-cell lymphoma was referred for a FDG PET/CT scan due to severe left lower abdominal pain to rule out recurrent cancer. These FDG PET/CT images and previous FDG PET/CT images 5 months ago both revealed an air bubble in the urinary bladder on the CT images. He had a recurrent urinary tract infection history for 6 months. A list-mode dynamic data acquisition with diuresis intravenous injection revealed linear FDG activity extending from the upper-left portion of the bladder to a soft tissue mass in the lower-left pelvic region. An enterovesical fistula was confirmed by surgery.

Foreign body in the ureter: a particle of glue after transarterial embolization of a renal pseudoaneurysm during percutaneous nephrostomy.

Reports on foreign bodies within the ureter are extremely rare in the literature. Herein, we present a case of a foreign body in a ureter, specifically a particle of glue resulting from transarterial embolization of a renal pseudoaneurysm secondary to percutaneous nephrostomy. Emergent transarterial embolization was required due to life-threatening active bleeding of the pseudoaneurysm. However, the glue material subsequently fell into the ureter where it became a foreign body, resulting in obstructive uropathy. Several surgical interventions, including endoscopic and laparoscopic methods, were performed to retrieve the foreign body, but these attempts were unsuccessful. Finally, the glue material was spontaneously passed out by chance. To the best of our knowledge, this type of complication (a glue particle left over from an embolization procedure migrating into the urinary collecting system) has never been reported. We recommend close follow-up examinations after transarterial embolization for renal pseudoaneurysm in order to avoid possible obstructive uropathy caused by glue materials or coils.

Changes in the endopolyploidy pattern of different tissues in diploid and tetraploid Phalaenopsis aphrodite subsp. formosana (Orchidaceae).

Endopolyploidy is frequently observed during development in plant species. Patterns of endopolyploidy are diverse in the various organs of different plant species. However, little is known about the role of endopolyploidization and its significance in orchids. This study was undertaken to determine the extent of endopolyploidy in different tissues of the diploid and tetraploid genotypes of Phalaenopsis aphrodite subsp. formosana and to examine the factors that contribute to increased ploidy levels. Endopolyploidy occurs in various tissues of diploid and tetraploid orchids, at different developmental stages and under different culture conditions, as determined by flow cytometry. In this study, different patterns of endopolyploidy were observed in parts of the protocorms, leaves, roots and flowers. Endopolyploidy was found in all tissues studied except the pollinia and the tetraploid ovaries. A higher degree of endopolyploidy was observed in mature tissues compared to young tissues, greenhouse-grown plants compared to in vitro plants and diploid plants compared to tetraploid plants. We discuss the relationships between endopolyploidization and several factors related to plant growth, as well as some practical considerations of these findings.

The suppressive effect of Rho kinase inhibitor, Y-27632, on oncogenic Ras/RhoA induced invasion/migration of human bladder cancer TSGH cells.

Urothelial cell carcinoma is the most common type of malignancy found in long-term dialysis patients and kidney transplant recipients in Taiwan. Surgical specimens of tumorous and non-tumorous bladder tissues were collected from 12 patients with bladder cancer. Increased expressions of Ras, RhoA, Akt, PI-3K were demonstrated in the tumors as compared to adjacent control tissues. To understand the impact of Ras over-expression on bladder cancer progression, human bladder cancer TSGH 8301 cells were transfected with Ras DNA. The Ras-transfected cells were then treated with either a PI-3K inhibitor (wortmannin) or Rho kinase inhibitor (Y-27632) and the expressions of Ras, PI-3K, Akt, NF-kappaB, and RhoA were analyzed. Fluorescent phalloidin staining demonstrated more intense F-actin staining in the Ras over-expressed cells than in the control cells, and the intensity of F-actin was inhibited by Y-27632. A gelatin zymography study demonstrated that the MMP-2 and MMP-9 expressions of the Ras-transfected cells were enhanced, and Y-27632 treatment reduced the levels of MMP-2 and MMP-9. Similarly, a wound healing assay revealed that the ability of cell migration was markedly increased by Ras transfection and the healing rate after treatment of Y-27632 was delayed. Our results provide evidence that Ras-induced RhoA and NF-kappaB activation was involved in the invasion/migration of bladder cancer. Through Ras and/or RhoA inhibition, there might be an opportunity for new therapeutic interventions in bladder cancer.

Multiple screening of urolithic organic acids with copper nanoparticle-plated electrode: potential assessment of urolithic risks.

There is yet to be a reliable prediction of urolithiasis. To facilitate early diagnosis, a simple and rapid high performance liquid chromatography method with electrochemical detection using disposable copper-nanoparticle-plated electrodes (Cu(n)-SPE) was developed for multiple detection of creatinine and 4 urolithic organic acids. A total of 206 normal and urolithic human and canine urines and urolith samples were collected for direct analysis of creatinine, cystine, uric acid, oxalic acid, and citric acid without sample cleanup and derivatization processes. Urinary organic acids were separated in 11 min and were devoid of ascorbic acid interference. The detection limits (S/N>3) were at the nanomolar level with linear dynamic ranges spanning 2-3 orders of magnitude. Recoveries in urine ranged from 99.5% for creatinine to 86.5% for citric acid. The analytical variations (RSD) were less than 6.2% in phosphate buffer and 7.7% in urine. Important differences in organic acid levels/profiles between animal species and among normal and urolithic urines/urolith were unveiled and corresponded well (70-90%) with the urolithic risk in a retrospective assessment. The simplicity and reproducibility of this method using disposable Cu(n)-SPE has made routine urine analysis possible and can be of great clinical and diagnostic potential in the screening of urolithiasis and abnormal states related to excess secretion of organic acids and amino acids in humans and animals.

Immunochemical and molecular characterization of GBC4 as a tanycyte-like cell line derived from grouper brain.

A clonal cell line, GBC4, derived from grouper (Epinephelus coioides) brain is proposed to represent an immature astroglial cell line because it expresses glial fibrillary acidic protein (GFAP), cytokeratin and vimentin. In teleost brain, tanycytes are the most abundant GFAP-expressing cell type, suggesting that GBC4 cells are derived from tanycytes. To test this hypothesis, protein and mRNA expression profiles of GBC4 cells were evaluated. We detected protein and/or mRNA expression of aromatase B, brain lipid binding protein, connexin43 protein, glutamine synthetase, S100 protein and Sox2. These proteins/mRNAs are also expressed in fish tanycytes. GBC4 cells also contained oligodendroglia proteins, including A2B5, galactocerebroside, myelin basic protein, proteolipid protein and platelet-derived growth factor receptor alpha as well as certain neuronal protein markers such as connexin35 protein and tyrosine hydroxylase. Our results indicate that GBC4 cells may be multipotent neural progenitor cells similar to tanycytes. Because GBC4 expresses several neural-specific genes, this line will be useful for studies on gene expression/regulation and neural development.

Effect of single botulinum toxin A injection to the external urethral sphincter for treating detrusor external sphincter dyssynergia in spinal cord injury.

OBJECTIVE: To evaluate the effect of a single cystoscopic injection of botulinum toxin to the external urethral sphincter in treating detrusor external sphincter dyssynergia. DESIGN: An open treatment trial with pre- and post-treatment evaluations. SUBJECTS: A total of 20 suprasacral spinal cord injured patients with pure detrusor external sphincter dyssynergia. METHODS: A single dose of 100 IU botulinum toxin A was applied into the external urethral sphincter via cystoscopy. Outcome measurements included maximal detrusor pressure, maximal urethral pressure, detrusor leak point pressure, integrated electromyography, and maximal pressure on static urethral pressure profilometry obtained 4 weeks post-treatment. Post-voiding residuals were checked pre- and post-injection. RESULTS: There were significant reductions in integrated electro notmyography and static and maximal urethral pressure, but not in maximal detrusor pressure and detrusor leak point pressure after treatment. Post-voiding residuals were significantly decreased at all evaluation periods. In the sub-group analysis, patients who showed good effects of treatment had significantly lower baseline integrated electro-myography (p<0.05). CONCLUSION: This study demonstrates the effect of a single cystoscopic injection of botulinum toxin in detrusor external sphincter dyssynergia. Integrated electromyography is a good evaluation tool for the net effect and dosage of botulinum toxin. Patients with severe spasticity over the external urethral sphincter may require repeated injections or higher doses.

Splenogonadal fusion.

Splenogonadal fusion (SGF) is a rare congenital non-malignant anomaly characterized by fusion of splenic tissue to the gonad, and can be continuous or discontinuous. Very few cases have been diagnosed preoperatively, and many patients who present with testicular swelling undergo unnecessary orchiectomy under the suspicion of testicular neoplasm. A 16-year-old boy presented with a left scrotal mass and underwent total excision of a 1.6-cm tumor without damaging the testis, epididymis or its accompanying vessels. Pathologic examination revealed SFG (discontinuous type). If clinically suspected before surgery, the diagnosis may be confirmed by Tc-99m sulfur colloid imaging, which shows uptake in both the spleen and accessory splenic tissue within the scrotum. Frozen section should be considered if there remains any doubt regarding the diagnosis during operation.

Acute anal stretch inhibits NMDA-dependent pelvic-urethra reflex potentiation via spinal GABAergic inhibition in anesthetized rats.

The impact of acute anal stretch on the pelvic-urethra reflex potentiation was examined in urethane-anesthetized rats by recording the external urethra sphincter electromyogram activity evoked by the pelvic afferent stimulation. Test stimulation (1 stimulation/30 s) evoked a baseline reflex activity with a single action potential that was abolished by gallamine (5 mg/kg iv). On the other hand, the repetitive stimulation (1 stimulation/1 s) induced spinal reflex potentiation (SRP) that was attenuated by intrathecal 6-cyano-7-nitroquinoxaline-2,4-dione (a glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionat receptor antagonist, 100 microM, 10 microl) and d-2-amino-5-phosphonovalerate [a glutamatergic N-methyl-D-aspartate (NMDA) antagonist, 100 microM, 10 microl]. Acute anal stretch using a mosquito clamp with a distance of 4 mm exhibited no effect, whereas distances of 8 mm attenuated and 12 mm abolished the repetitive stimulation-induced SRP. Intrathecal NMDA (100 microM, 10 microl) reversed the abolition on SRP caused by anal stretch. On the other hand, pretreated bicuculline [gamma-aminobutyric acid (GABA) A receptor antagonist, 100 microM, 10 microl] but not hydroxysaclofen (GABAB receptor antagonist) counteracted the abolition on the repetitive stimulation-induced SRP caused by the anal stretch. All of the results suggested that anal stretch may be used as an adjunct to assist voiding dysfunction in patients with overactive urethra sphincter and that GABAergic neurotransmission is important in the neural mechanisms underlying external urethra sphincter activity inhibited by anal stretch.

Gossypiboma--retained surgical sponge.

Intra-abdominal retained surgical sponge is an uncommon surgical error. Herein, we report a 92-year-old woman who was brought to the emergency room for acute urinary retention. She had a history of vaginal hysterectomy for uterine prolapse 18 years previously, performed at our hospital. Retained surgical sponge in the pelvic cavity was suspected by abdominal computed tomography. The surgical gauze was removed by laparotomy excision and the final diagnosis was gossypiboma.

Signal peptide of eosinophil cationic protein upregulates transforming growth factor-alpha expression in human cells.

Eosinophil cationic protein (ECP) is a major component of eosinophil granule protein that is used as a clinical bio-marker for asthma and allergic inflammatory diseases. Previously, it has been reported that the signal peptide of human ECP (ECPsp) inhibits the cell growth of Escherichia coli (E. coli) and Pichia pastoris (P. pastoris), but not mammalian A431 cells. The inhibitory effect is due to the lack of human signal peptide peptidase (hSPP), a protease located on the endoplasmic reticulum (ER) membrane, in the lower organisms. In this study, we show that the epidermal growth factor receptor (EGFR) is upregulated by the exogenous ECPsp-eGFP as a result of the increased expression of the transforming growth factor-alpha (TGF-alpha) at both transcriptional and translational levels in A431 and HL-60 clone 15 cell lines. Furthermore, the N-terminus of ECPsp fragment generated by the cleavage of hSPP (ECPspM1-G17) gives rise to over threefold increase of TGF-alpha protein expression, whereas another ECPsp fragment (ECPspL18-A27) and the hSPP-resistant ECPsp (ECPspG17L) do not show similar effect. Our results indicate that the ECPspM1-G17 plays a crucial role in the upregulation of TGF-alpha, suggesting that the ECPsp not only directs the secretion of mature ECP, but also involves in the autocrine system.

Activation of Rac1 by phosphatidylinositol 3-kinase in vivo: role in activation of mitogen-activated protein kinase (MAPK) pathways and retinoic acid-induced neuronal differentiation of SH-SY5Y cells.

Rho GTPases such as RhoA, Rac1 and Cdc42 are crucial players in the regulation of signal transduction pathways required for neuronal differentiation. Using an in vitro cell culture model of neuroblastoma SH-SY5Y cells, we demonstrated previously that RhoA is an in vivo substrate of tissue transglutaminase (TGase) and retinoic acid (RA) promoted activation of RhoA by transamidation. Although activation of RhoA promoted cytoskeletal rearrangement in SH-SY5Y cells, it was not involved in induction of neurite outgrowth. Here, we demonstrate that RA promotes activation of Rac1 in SH-SY5Y cells in a transamidation-independent manner. RA-induced activation of Rac1 is mediated by phosphatidylinositol 3-kinase (PI3K), probably because of phosphorylation of the p85 regulatory subunit by Src kinases. Over-expression of constitutively active PI3K or Rac1-V12 induces neurite outgrowth, activation of mitogen activated protein kinases (MAPKs), and expression of neuronal markers. The PI3K inhibitor LY294002, or over-expression of dominant negative Rac1-N17, blocks RA-induced neurite outgrowth, activation of MAPKs, and expression of neuronal markers, suggesting that activation of PI3K/Rac1 signaling represents a potential mechanism for regulation of neuronal differentiation in SH-SY5Y cells.