Epidermal growth factor-induced ANGPTL4 enhances anoikis resistance and tumour metastasis in head and neck squamous cell carcinoma.

Epidermal growth factor (EGF) is important for cancer cell proliferation, angiogenesis and metastasis in many types of cancer. However, the mechanisms involved in EGF-induced head and neck squamous cell carcinoma (HNSCC) metastasis remain largely unknown. In this study, we reveal that angiopoietin-like 4 (ANGPTL4) plays an important role in the regulation of EGF-induced cancer metastasis. We showed that EGF-induced ANGPTL4 expression promoted anoikis resistance and cancer cell migration and invasion in HNSCC. In addition, depletion of ANGPTL4 inhibited EGF-induced cancer cell invasion. Autocrine production of EGF-induced ANGPTL4 regulated the expression of matrix metalloproteinases (MMPs). The induction of MMP-1 gene expression by ANGPTL4-activated integrin ß1 signalling occurred through the AP-1 binding site in the MMP-1 gene promoter. Furthermore, down-regulation of MMP-1 impeded EGF- and recombinant ANGPTL4-enhanced HNSCC cell migration and invasion. Depletion of ANGPTL4 significantly blocked EGF-primed extravasation and metastatic seeding of tumour cells and MMP-1 expression in lungs. However, no effect of ANGPTL4 on tumour growth was observed. These results suggest that EGF-induced expression and autocrine production of ANGPTL4 enhances HNSCC metastasis via the up-regulation of MMP-1 expression. Inhibition of ANGPTL4 expression may be a potential strategy for the treatment of EGFR-mediated HNSCC metastasis.

Cell-to-Cell Spread of HIV and Viral Pathogenesis.

Human immunodeficiency virus type 1 (HIV-1) gives rise to a chronic infection that progressively depletes CD4(+) T lymphocytes. CD4(+) T lymphocytes play a central coordinating role in adaptive cellular and humoral immune responses, and to do so they migrate and interact within lymphoid compartments and at effector sites to mount immune responses. While cell-free virus serves as an excellent prognostic indicator for patient survival, interactions of infected T cells or virus-scavenging immune cells with uninfected T cells can greatly enhance viral spread. HIV can induce interactions between infected and uninfected T cells that are triggered by cell surface expression of viral Env, which serves as a cell adhesion molecule that interacts with CD4 on the target cell, before it acts as the viral membrane fusion protein. These interactions are called virological synapses and promote replication in the face of selective pressure of humoral immune responses and antiretroviral therapy. Other infection-enhancing cell-cell interactions occur between virus-concentrating antigen-presenting cells and recipient T cells, called infectious synapses. The exact roles that these cell-cell interactions play in each stage of infection, from viral acquisition, systemic dissemination, to chronic persistence are still being determined. Infection-promoting immune cell interactions are likely to contribute to viral persistence and enhance the ability of HIV-1 to evade adaptive immune responses.

Phenotype and function of nasal dendritic cells.

Intranasal (i.n.) vaccination generates immunity across local, regional, and distant sites. However, nasal dendritic cells (DCs), pivotal for the induction of i.n. vaccine-induced immune responses, have not been studied in detail. Here, by using a variety of parameters, we define nasal DCs in mice and humans. Distinct subsets of "classical" DCs, dependent on the transcription factor zbtb46 were identified in the murine nose. The murine nasal DCs were Fms-related tyrosine 3 kinase ligand responsive and displayed unique phenotypic and functional characteristics, including the ability to present antigen, induce an allogeneic T-cell response, and migrate in response to lipopolysaccharide or live bacterial pathogens. Importantly, in a cohort of human volunteers, BDCA-1(+) DCs were observed to be the dominant nasal DC population at steady state. During chronic inflammation, the frequency of both BDCA-1(+) and BDCA-3(hi) DCs was reduced in the nasal tissue, associating the loss of these immune sentinels with chronic nasal inflammation. The present study is the first detailed description of the phenotypic, ontogenetic, and functional properties of nasal DCs, and will inform the design of preventative immunization strategies as well as therapeutic modalities against chronic rhinosinusitis.

Residual effect of reductions in red blood cell count and haematocrit and haemoglobin levels after 10-month withdrawal of pioglitazone in patients with Type 2 diabetes.

AIM: To investigate the recovery of thiazolidinedione-induced body weight gain and haematopoietic changes after stopping pioglitazone treatment in patients with Type 2 diabetes. METHODS: This retrospective cohort study included 214 patients divided into three groups according to pioglitazone treatment status. The first study arm included patients who received pioglitazone for 38 months then interrupted this for 10 months (pioglitazone-interruption group). The second arm consisted of patients who received pioglitazone throughout the 48 months (pioglitazone-continuous group); the third arm included patients who had never received pioglitazone therapy (control group). RESULTS: Red blood cell count and haematocrit and haemoglobin levels decreased significantly, while body weight increased in the two pioglitazone-treated groups as compared with the control group at 38 months. Multivariate regression analysis showed that the reductions in red blood cell count/haemoglobin levels were associated with pioglitazone use. In the pioglitazone-interruption group, no recoveries of red blood cells, or haematocrit or haemoglobin levels were observed after stopping pioglitazone for 10 months compared with the pioglitazone-continuous group, but body weight gain decreased to a level that was significantly lower than that in the pioglitazone-continuous group and did not differ significantly from the control group. CONCLUSION: In this study, we observed a reversal of body weight gain but no recoveries in red blood cells or haematocrit or haemoglobin levels after stopping pioglitazone for 10 months in patients treated with pioglitazone for 38 months. This finding should prompt a reconsideration of the sustained effect of thiazolidinediones on the haematopoietic system in patients with Type 2 diabetes.

Improved axonal regeneration of transected spinal cord mediated by multichannel collagen conduits functionalized with neurotrophin-3 gene.

Functionalized biomaterial scaffolds targeted at improving axonal regeneration by enhancing guided axonal growth provide a promising approach for the repair of spinal cord injury. Collagen neural conduits provide structural guidance for neural tissue regeneration, and in this study it is shown that these conduits can also act as a reservoir for sustained gene delivery. Either a G-luciferase marker gene or a neurotrophin-3-encoding gene, complexed to a non-viral, cyclized, PEGylated transfection vector, was loaded within a multichannel collagen conduit. The complexed genes were then released in a controlled fashion using a dual release system both in vitro and in vivo. For evaluation of their biological performance, the loaded conduits were implanted into the completely transected rat thoracic spinal cord (T8-T10). Aligned axon regeneration through the channels of conduits was observed one month post-surgery. The conduits delivering neurotrophin-3 polyplexes resulted in significantly increased neurotrophin-3 levels in the surrounding tissue and a statistically higher number of regenerated axons versus the control conduits (P<0.05). This study suggests that collagen neural conduits delivering a highly effective non-viral therapeutic gene may hold promise for repair of the injured spinal cord.

Socioeconomic correlates of inpatient spending for patients with type 2 diabetes mellitus in China: evidence from Hangzhou.

AIMS: We evaluated the factors associated with inpatient costs including total costs, pharmaceutical costs and laboratory costs for diabetes-related admissions. PATIENTS AND METHODS: Using data for 960 adult patients admitted between May 2005 and April 2008 with a primary or secondary diagnosis of type 2 diabetes mellitus (DM) at Sir Run Run Shaw Hospital affiliated with Zhejiang University Medical School (SRRSH) in Hangzhou, China, we evaluate the association between patient characteristics and inpatient costs with multivariable regression analyses. RESULTS: Total inpatient costs were positively associated with age, higher UKPDS stroke risk score, and presence of any complication. A regression that included patient socioeconomic and clinical characteristics explained 21.5% of the variation in total inpatient costs; regression estimates indicate that patients with coronary artery disease, retinopathy, nephropathy, neuropathy, and diabetic foot had inpatient costs that were respectively 93.7%, 14.0%, 17.5%, 11.5% and 89.0% higher than otherwise similar patients without those complications. Pharmaceutical costs did not differ by insurance coverage. Insured patients spent 7-16% more on laboratory tests than otherwise similar patients did. CONCLUSIONS: Clinical factors, especially presence of diabetes-related complications, appear to be the primary determinants of variation in inpatient costs for patients with type 2 DM in China. To mitigate the health costs increases associated with China's DM epidemic, policymakers should focus on cost-effective ways to manage patients in outpatient settings to prevent the complications associated with diabetes.

Stability and nonlinear optical properties of ZnO nanoparticles.

Photoluminescence (PL) of ZnO nanoparticles of different surface states and sizes grown by several methods has been measured. The origin of luminescence and dependence of the luminescence spectrum shape and intensity on 325 nm excitation laser power are studied. Strong ultraviolet emission at 3.26 eV, weak violet emission around 3.12 eV and weak green emission at 2.40 eV have been observed in 16 nm nanoparticles capped by octylamine grown by non-hydrolytic method. The nanoparticles are stable under high power laser radiation and their PL intensity increases nonlinearly with an increasing laser power. As the nanoparticle size decreases to 12 nm, high power laser produces nonradiative centers which may quench the luminescence in a degree. Nanoparticles of 8 nm capped by PVP and uncapped nanoparticles of 14 nm are unstable and their luminescence depends on the excitation laser power. High power laser can quench O vacancy emission and enhance ultraviolet emission in PVP capped nanoparticles while vacancy emission can not be quenched in uncapped nanoparticles.

CdO/ZnO core/shell nanostructures grown from single molecular precursors.

Quadrilateral CdO nanoparticles were grown from cadmium cupferronate complex by injecting the precursor dissolved in octylamine into trioctylamine at 220 and 250 degrees C. CdO/ZnO core/shell structures were synthesized with a method similar to that of growing CdO cores by injecting the shell precursor following the growth of cores. The shell growth temperature was adjusted from 160 to 130 degrees C, and the shell precursor supply speed was adjusted from 4 to 8 ml/h. The obtained nanostructures were characterized by X-ray diffraction, high resolution transmission electron microscope, and ultraviolet-visible spectrophotometer. No core/shell structures formed if the shell precursor injection speed was as high as 8 ml/h. A very thin layer of ZnO shells would form on CdO cores if the shell precursor was injected at a speed of 4 ml/h at temperature of 160 degrees C, and the shells had good crystal quality. CdO/ZnO core/shell nanostructures were inclined to be spherical, and no homogeneous formation of ZnO nanoparticles was observed if the shell precursor injection temperature was lowered to 130 degrees C.

Calcineurin-mediated dephosphorylation of c-Jun Ser-243 is required for c-Jun protein stability and cell transformation.

The proto-oncogene c-Jun plays an important role in regulating tumor progression. We previously reported that the serine/threonine phosphatase calcineurin (CaN, also called PP2B) dephosphorylates the C-terminus (Ser-243) of c-Jun, resulting in the increase in c-Jun and Sp1 interaction, and subsequent c-Jun-induced gene expression. Here, we demonstrate the interaction of c-Jun and CaN in the nucleus of living cells by fluorescence resonance energy transfer assay and that this interaction is mediated through the calmodulin-binding domain of CaN. Furthermore, c-Jun protein stability was altered by CaN-mediated dephosphorylation at the Ser-243 site of c-Jun. The half-life of the c-Jun mutant, c-Jun-S243A was longer than that of the wild-type c-Jun. Moreover, silencing of endogenous CaN expression led to increased c-Jun ubiquitination and decreased stability. In 46% of clinical cervical tissue samples obtained from patients with cervical cancer, enhanced c-Jun and CaN expression, as well as decreased phospho-Ser-243 expression levels were detected. Our results suggest that CaN stabilizes c-Jun by dephosphorylating c-Jun at Ser-243 to enhance its tumorigenic ability.

Role of transcriptional factors Sp1, c-Rel, and c-Jun in LPS-induced C/EBPdelta gene expression of mouse macrophages.

Transcription factor C/EBPs are involved in the regulation of various cellular responses. Here, it was suggested that C/EBPdelta gene was activated by lipopolysaccharide (LPS) through transcription factors Sp1, c-Rel, and c-Jun. Assay of the luciferase reporter vectors containing a 5'-deletion of the C/EBPdelta gene promoter indicated that a LPS-responsive element was positioned between -345 and -35 bp of mouse C/EBPdelta gene promoter. Transcription factors Sp1, c-Rel, and c-Jun bound to this region were identified using both in vivo chromatin immunoprecipitation and in vitro DNA-protein binding assays. LPS enhanced the proteins and DNA binding capacities of c-Rel and c-Jun, and the downstream Sp1 site was essential for LPS-induced C/EBPdelta gene. Treatment of cells with ERK/JNK/p38 inhibitors or NF-kappaB inhibitor inhibited the LPS-induced C/EBPdelta gene expression by inhibiting c-Jun, c-Rel, and p300 binding to DNA. Our findings provide a better understanding of LPS-induced C/EBPdelta gene expression.

3D finite element analyses of insertion of the Nucleus standard straight and the Contour electrode arrays into the human cochlea.

Previous experimental studies of insertion of the Nucleus standard straight and the Contour arrays into the scala tympani have reported that the electrode arrays cause damage to various cochlear structures. However, the level of insertion-induced damage by these electrode arrays to cochlear structures (the spiral ligament, the basilar membrane and the osseous spiral lamina) has not been quantified. Although it has been suggested that rotation can overcome this resistance and prevent the basilar membrane from being pierced by the tip of the Nucleus standard straight array, there has not been any attempt to study the relationship between the rotation and the reduction of damage to the basilar membrane. In this study, 3D finite element analyses of insertions of the Nucleus standard straight array and the Contour array into the scala tympani have been undertaken. The perforation of the basilar membrane by the tip of the Nucleus standard straight array at the region of 11-14 mm from the round window appears to be compounded by the geometry of the spiral passage of the scala tympani. Anti-clockwise rotations between 25 degrees and 90 degrees applied at the basal end of the electrode array (for the right cochlea) were shown to significantly reduce the contact stresses exerted by the tip on the basilar membrane which support the practice of applying small rotation partway through insertion of electrode array to minimize damage to the basilar membrane. Although the Contour array (with its stylet intact) is stiffer than the Nucleus standard straight array, a slight withdrawal of the stylet from the Contour array before insertion was found to significantly reduce damage by the electrode array to the spiral ligament and the basilar membrane.

Determination of frictional conditions between electrode array and endosteum lining for use in cochlear implant models.

Frictional conditions between the electrode array (in cochlear implants) and the endosteum lining covering the walls of the interior scala tympani structure strongly influence the sliding behaviour of the electrode array. Friction coefficients, determined by a simple but effective method based on the impending slippage model of electrode arrays sliding over the endosteum lining are reported in this paper. In this study, friction coefficients of the Nucleus standard straight and the Contour arrays have been determined with and without lubricants applied on the endosteum lining. In the absence of applied lubricants, friction coefficients were found to be 0.19 for the Nucleus standard straight array and 0.12 for the Contour array. Application of lubricants (glycerin and sorbelene) has the potential to lower the friction coefficient for Nucleus standard straight array (0.12 and 0.15) and for the Contour array (0.04 and 0.08). These results are used in finite element models to predict accurately the trajectories of electrode arrays and sliding contact pressures on cochlear structures to evaluate the likelihood of damage sustained during insertion.

Stiffness properties for Nucleus standard straight and contour electrode arrays.

Trauma and damage during insertion of electrode arrays into the human cochlea are strongly related to the stiffness of the array. The stiffness properties of electrode arrays, which were determined by three-point flexural bending and buckling tests, are reported in this paper. To date there has been limited publication on mechanical properties of these electrode arrays. Previous studies mainly focused on characterizing the stiffness of the tip of the Nucleus straight array with little emphasis on characterizing the stiffness of its whole length. In this study, stiffnesses of the Nucleus straight and contour electrode arrays have been determined along their length. Young's modulus of elasticity of the Nucleus straight array has been found to increase from the tip (182 MPa) to the rear end (491 MPa), whereas the stiffness of the contour array is greatest near the tip (480 MPa) and is fairly uniform in the middle and rear sections of the electrode array (380-400 MPa). Buckling experiments have shown that the contour array has much higher critical buckling load (about four times) than the Nucleus straight array. The results from three-point flexural bending and buckling experiments provide significant data for the development of electrode arrays, from which new array designs with improved flexibility can be developed. The results of stiffness properties are also important input for use in finite element models to predict the trajectories during insertion and to help evaluate the effects of different electrode array designs on damage sustained during insertion.

Evaluation of trajectories and contact pressures for the straight nucleus cochlear implant electrode array - a two-dimensional application of finite element analysis.

A two-dimensional (2D) finite element analysis has been used in this study to model the insertion of the Nucleus electrode array with different stiffness properties in order to evaluate the propensity of damage by visualizing the predicted trajectories and by comparing the buckling stresses and the contact pressures at the tip (and its distribution along the length) of the electrode array. Previous temporal bone studies have shown that damage during insertion of an electrode array around the basal turn of the cochlear spiral could be related to the design and the stiffness properties of the electrode array. However, it is difficult to evaluate different designs of electrode arrays purely by experimental methods as the experimental conditions and their results are difficult to reproduce. Three electrode arrays with different mechanical properties, i.e. uniform stiffness, graded stiffness, and a soft tip have been modelled. Buckling stress and contact pressure at the tip of the electrode array were found to be highest for the arrays with uniform stiffness. The contact pressures at the tip of the electrode array appeared strongly influenced by the stiffness profile and were optimal for graded stiffness. The results indicate the importance of the electrode array design and stiffness properties in minimizing trauma. However, there are a number of limitations in the present 2D evaluation which will require further analysis using a three-dimensional model to obtain definitive results.

Efficient assembly of an HIV-1/MLV Gag-chimeric virus in murine cells.

In human cells infected by HIV type 1 (HIV-1), the viral Gag protein directs the assembly of nascent viral particles at the plasma membrane. In murine cells, HIV-1 Gag fails to reach the plasma membrane and instead forms nonfunctional intracellular aggregates. The viral determinants of this species incompatibility are previously undefined. To address this problem, we replaced a region of HIV-1 Gag known to direct its localization, the matrix (MA) domain, with functionally homologous regions from Moloney murine leukemia virus (MLV), a murine retrovirus. An HIV-1 clone carrying such a chimeric Gag protein, designated murine HIV (MHIV), assembled more efficiently than nonchimeric HIV-1 and restored plasma membrane localization of Gag in murine cells. Increased efficiency of viral assembly in murine cells was observed from MHIV constructs carrying MLV MA in place of HIV-1 MA. Efficient processing of the HIV-1 capsid protein from the chimeric Gag polyprotein and subsequent infectivity of MHIV required the presence of MLV p12 in addition to MLV MA. These findings strongly suggest that the HIV-1 MA domain of HIV-1 Gag is responsible for the assembly defect in mouse cells. Although these MHIV do not recruit native HIV-1 Env efficiently, they are capable of single-round infection when produced by high-efficiency transfection of human 293 cells and provided with an HIV-1 Env lacking its cytoplasmic tail. With further adaptation, this chimeric MHIV approach may provide the basis for creating an infectious mouse model for HIV/AIDS.

Topical anesthetics in children: agents and techniques that equally comfort patients, parents, and clinicians.

Topical anesthetics are increasingly important, as the number of outpatient surgeries for dermatologic problems in infants and children is steadily growing. This noninvasive modality of anesthetic delivery in conjunction with a reassuring environment may minimize the discomfort of otherwise painful procedures. Since the 1880s, when cocaine was first used as a topical ophthalmologic anesthetic, many ester-and amide-based local anesthetics have been developed for a variety of simple and complex procedures. The pediatric dermatologist's arsenal of topical anesthetic preparations is increasing with the development of novel vehicles of transdermal delivery and the use of anesthetics in combination. Eutectic mixture of local anesthetics is currently the most frequently prescribed topical agent, though the use of ELA-max, another lidocaine-containing preparation, is gaining momentum, especially in the neonatal population. Amethocaine, tetracaine, iontophoresis, and the S-caine patch, a product on the horizon for use in the pediatric population, also are included in this discussion.

Tinea capitis update: a continuing conflict with an old adversary.

Infection with tinea capitis in childhood is a common, age-old problem that continues to plague patients and their families. As is true for most infectious diseases, the epidemiology of tinea capitis is in a constant state of flux and varies considerably with respect to geography and specific patient populations. Trichophyton tonsurans is now the most common cause of tinea capitis in the United States. A recent epidemiologic observation is a striking increase in the incidence of tinea capitis, particularly among African-Americans. Clinical studies over the past decade that have investigated the response of tinea capitis to griseofulvin, the mainstay treatment for this condition, suggest a decrease in sensitivity to this pharmacologic agent, in association with this new epidemiology. Important advances in the diagnosis and treatment of tinea capitis include a renewed interest in the use of the cotton swab method of diagnosing fungal cultures in children, and the ongoing investigation of promising new medications for the treatment of tinea capitis, including terbinafine, itraconazole, and fluconazole in this era of resistant organisms.

Symptomatic post-traumatic cyst of the liver: treatment by laparoscopic surgery.

We report a case of a large post-traumatic liver cyst in a symptomatic patient treated by laparoscopic excision in an ambulatory setting. This rare lesion can be treated safely by this alternative modality on an outpatient basis, with minimal morbidity.

Pediatric anesthesia in dermatologic surgery: when hand-holding is not enough.

BACKGROUND: Dermatologic procedures in children may require the use of topical and local anesthetics, sedatives, and general anesthesia. OBJECTIVE: To review developments in topical and local anesthetics, sedatives, and general anesthesia relevant to dermatologic procedures in children. METHODS: Review of the medical literature. RESULTS: Topical anesthetics, including EMLA and liposome-encapsulated lidocaine cream, amethocaine, cetacaine, and benzocaine products may be useful for decreasing the pain of cutaneous procedures including intra-lesional lidocaine infiltration. A variety of sedative and hypnotic agents may be utilized for pediatric dermatology procedures, and guidelines for their appropriate use have been published. General anesthesia for dermatologic procedures in the pediatric population is appropriate for a variety of procedures including laser treatment of capillary malformations. CONCLUSION: A variety of anesthetic, analgesic, and sedatives may be useful for pediatric cutaneous surgery.

Palmitoylation of the HIV-1 envelope glycoprotein is critical for viral infectivity.

Recent studies suggest that HIV-1 budding occurs selectively from detergent-insoluble membrane domains, referred to as lipid rafts. Palmitoylation is thought to be one of the factors responsible for targeting membrane proteins to lipid rafts. The cytoplasmic domain of the HIV-1 envelope glycoprotein (gp160) contains two palmitoylated cysteine residues. In this work, we studied the solubility of gp160 after detergent extraction. We show that wild-type gp160 is mostly insoluble after ice-cold Triton X-100 extraction, but that it becomes almost completely soluble at 37 degrees C. In contrast, we find that a mutant gp160, in which the two palmitoylated cysteine residues are replaced by serine, is Triton X-100 soluble even under ice-cold extraction. These findings are consistent with the properties of proteins that localize to lipid rafts and strongly suggest that gp160 is associated with lipid rafts. Further, removal of both palmitoylation sites results in the formation of virus with low levels of gp160 incorporation as well as a decrease in viral infectivity by 60-fold. Our results strongly support the suggestion that HIV-1 buds from lipid rafts and point to a role for rafts as a viral assembly hub.