bluuebunny/crossref_metadata_2025 huggingface dataset:
Peripheral nerve regeneration using composite poly(lactic acid‐caprolactone)/nerve growth factor conduits prepared by coaxial electrospinning
Authors: Liu Jun‐Jian , Wang Chun‐Yang , Wang Jian‐Guang , Ruan Hong‐Jiang , Fan Cun‐Yi
Published: October 2010

Many neurotrophic factors have been shown to promote neurite outgrowth by improving the microenvironment that is required for nerve regeneration. However, the delivery of these bioactive agents to the nerve injury site, as well as effective and local release, remains a challenging problem. We have developed a novel composite nerve conduit comprised of poly(lactic acid‐caprolactone) (P(LLA‐CL)) and nerve growth factor (NGF). This was developed from core‐shell structured biodegradable nanofibers, which were fabricated by coaxial electrospinning of P(LLA‐CL) for the shell and bovine serum albumin (BSA) or BSA/NGF for the core. In rats, gaps of 10‐mm long sciatic nerves were bridged using an autograft, an empty P(LLA‐CL) conduit, a NGF injection P(LLA‐CL) conduit, a P(LLA‐CL)/NGF composite conduit, respectively. Regenerated nerve fibers were harvested and morphological and functional evaluation of nerve regeneration was performed at 12 weeks postsurgery. Although partial biodegradation and small cracks in the conduits were observed, the conduit outlines remained intact for 12 weeks after surgery. Based on functional and histological observations, the number and arrangement of regenerated nerve fibers, myelination, and nerve function reconstruction was similar in the P(LLA‐CL)/NGF conduit group to that of the nerve autograft group (p > 0.05), but was significantly greater to the empty P(LLA‐CL) and injection NGF P(LLA‐CL) conduit groups (both p < 0.05). Therefore, the composite P(LLA‐CL)/NGF conduit, which exhibited favorable mechanical properties and biocompatibility, could effectively promote sciatic nerve regeneration in rats. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.
Chondrogenesis in perfusion bioreactors using porous silk scaffolds and hESC‐derived MSCs
Authors: Tığlı R. Seda , Cannizaro Chris , Gümüşderelioğlu Menemşe , Kaplan David L.
Published: October 2010

Tissue engineered cartilage can be grown in vitro with the use of cell‐scaffold constructs and bioreactors. The present study was designed to investigate the effects of perfusion bioreactors on the chondrogenic potential of engineered constructs prepared from porous silk fibroin scaffolds seeded with human embryonic stem cell (hESC)‐derived mesencyhmal stem cells (MSCs). After four weeks of incubation, constructs cultured in perfusion bioreactors showed significantly higher amounts of glycosaminoglycans (GAGs) (p < 0.001), DNA (p < 0.001), total collagen (p < 0.01), and collagen II (p < 0.01) in comparison to static culture. Mechanical stiffness of constructs increased 3.7‐fold under dynamic culture conditions and RT‐PCR results concluded that cells cultured in perfusion bioreactors highly expressed (p < 0.001) cartilage‐related genes when compared with static culture. Distinct differences were noted in tissue morphology, including polygonal extracellular matrix structure of engineered constructs in thin superficial zones and an inner zone under static and dynamic conditions, respectively. The results suggest that the utility of perfusion bioreactors to modulate the growth of tissue‐engineered cartilage and enhance tissue growth in vitro. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.
Wear behavior and in vitro cytotoxicity of wear debris generated from hydroxyapatite–carbon nanotube composite coating
Authors: Lahiri Debrupa , Benaduce Ana Paula , Rouzaud Francois , Solomon Jonathan , Keshri Anup Kumar , Kos Lidia , Agarwal Arvind
Published: October 2010

This work evaluates the effect of carbon nanotube (CNT) addition to plasma‐sprayed hydroxyapatite (HA) coating on its tribological behavior, biocompatibility of the coating, and cytotoxicity of CNT‐containing wear debris. Biological response of the CNT‐containing wear debris is critical for osteoblasts, the bone‐forming cells, and macrophages, the cells that clear up wear debris from blood stream. The addition of 4 wt % CNTs to HA coating reduces the volume of wear debris generation by 80% because of the improved elastic modulus and fracture toughness. CNT reinforcement has a pronounced effect on the particle size in the wear debris and subsequent biological response. There was a slight increase in the numbers and viability of osteoblasts grown on HA–CNT compared with HA alone. The cytotoxic effect of HA and HA–CNT debris to macrophages and osteoblasts was similar, demonstrating that loose CNT does not pose a problem to these cells. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.
Milk intake and risk of hip fracture in men and women: A meta-analysis of prospective cohort studies
Authors: Bischoff-Ferrari Heike A , Dawson-Hughes Bess , Baron John A , Kanis John A , Orav Endel J , Staehelin Hannes B , Kiel Douglas P , Burckhardt Peter , Henschkowski Jana , Spiegelman Donna , Li Ruifeng , Wong John B , Feskanich Diane , Willett Walter C
Published: October 2010

Milk contains calcium, phosphorus, and protein and is fortified with vitamin D in the United States. All these ingredients may improve bone health. However, the potential benefit of milk on hip fracture prevention is not well established. The objective of this study was to assess the association of milk intake with risk of hip fracture based on a meta-analysis of cohort studies in middle-aged or older men and women. Data sources for this study were English and non-English publications via Medline (Ovid, PubMed) and EMBASE search up to June 2010, experts in the field, and reference lists. The idea was to compare prospective cohort studies on the same scale so that we could calculate the relative risk (RR) of hip fracture per glass of milk intake daily (approximately 300 mg calcium per glass of milk). Pooled analyses were based on random effects models. The data were extracted by two independent observers. The results show that in women (6 studies, 195,102 women, 3574 hip fractures), there was no overall association between total milk intake and hip fracture risk (pooled RR per glass of milk per day = 0.99; 95% confidence interval [CI] 0.96–1.02; Q-test p = .37). In men (3 studies, 75,149 men, 195 hip fractures), the pooled RR per daily glass of milk was 0.91 (95% CI 0.81–1.01). Our conclusion is that in our meta-analysis of cohort studies, there was no overall association between milk intake and hip fracture risk in women but that more data are needed in men. © 2011 American Society for Bone and Mineral Research.
A minimal implementation of the AMBER–GAUSSIAN interface for ab initio QM/MM‐MD simulation
Authors: Okamoto Takuya , Yamada Kenta , Koyano Yoshiyuki , Asada Toshio , Koga Nobuaki , Nagaoka Masataka
Published: October 2010

For applying to a number of theoretical methodologies based on an ab initio quantum mechanical/molecular mechanical (QM/MM) molecular dynamics method connecting AMBER9 with GAUSSIAN03, we have developed an AMBER–GAUSSIAN interface (AG‐IF), which can be one of the simplest architectures. In the AG‐IF, only a few subroutines addition is necessary to retrieve the QM/MM energy and forces, obtained by GAUSSIAN, for solving a set of Newtonian equations of motion in AMBER. It is, therefore, easy to be modified for individual applications since AG‐IF utilizes most of those functions originally equipped not only in AMBER but also in GAUSSIAN. In the present minimal implementation, only AMBER is modified, whereas GAUSSIAN is left unchanged. Moreover, a different method of calculating electrostatic forces of MM atoms interacting with QM region is proposed. Using the AG‐IF, we also demonstrate three examples of application: (i) the QM versus MM comparison in the radial distribution function, (ii) the free energy gradient method, and (iii) the charge from interaction energy and forces. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2011
Theoretical study on the kinetics of OH radical reactions with CH3OOH and CH3CH2OOH
Authors: Luo Jie , Jia Xiujuan , Gao Yang , Song Guicai , Yu Yanbo , Wang Rongshun , Pan Xiumei
Published: October 2010

The mechanisms and kinetics studies of the OH radical with alkyl hydroperoxides CH3OOH and CH3CH2OOH reactions have been carried out theoretically. The geometries and frequencies of all the stationary points are calculated at the UBHandHLYP/6‐311G(d,p) level, and the energy profiles are further refined by interpolated single‐point energies method at the MC‐QCISD level of theory. For two reactions, five H‐abstraction channels are found and five products (CH3OO, CH2OOH, CH3CH2OO, CH2CH2OOH, and CH3CHOOH) are produced during the above processes. The rate constants for the CH3OOH/CH3CH2OOH + OH reactions are corrected by canonical variational transition state theory within 250–1500 K, and the small‐curvature tunneling is included. The total rate constants are evaluated from the sum of the individual rate constants and the branching ratios are in good agreement with the experimental data. The Arrhenius expressions for the reactions are obtained. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2011
The temperature intervals with global exchange of replicas empirical accelerated sampling method: Parameter sensitivity and extension to a complex molecular system
Authors: Li Xianfeng , Latour Robert A.
Published: October 2010

The recently developed “temperature intervals with global exchange of replicas” (TIGER2) algorithm is an efficient replica‐exchange sampling algorithm that provides the freedom to specify the number of replicas and temperature levels independently of the size of the system and temperature range to be spanned, thus making it particularly well suited for sampling molecular systems that are considered to be too large to be sampled using conventional replica exchange methods. Although the TIGER2 method is empirical in nature, when appropriately applied it is able to provide sampling that satisfies the balance condition and closely approximates a Boltzmann‐weighted ensemble of states. In this work, we evaluated the influence of factors such as temperature range, temperature spacing, replica number, and sampling cycle design on the accuracy of a TIGER2 simulation based on molecular dynamics simulations of alanine dipeptide in implicit solvent. The influence of these factors is further examined by calculating the properties of a complex system composed of the B1 immunoglobulin‐binding domain of streptococcal protein G (protein G) in aqueous solution. The accuracy of a TIGER2 simulation is particularly sensitive to the maximum temperature level selected for the simulation. A method to determine the appropriate maximum temperature level to be used in a TIGER2 simulation is presented. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2011
Myoepithelioma of the hand and carpal tunnel: An unusual cause of median nerve compression
Authors: Zamora Carlos Armando , Zamora Moisés Armando , Soto José Domingo , Garcés Miguel Ángel
Published: October 2010

Myoepitheliomas are rare tumors increasingly recognized to occur in the soft tissues. Although the hand and carpal tunnel are exceptional locations, the presence of these lesions in such sites constitutes a potential cause of debilitating symptoms. We report the case of a patient with severe pain secondary to median nerve compression and displacement of flexor tendons caused by a rapidly growing myoepithelioma. This is the first sonographic description of this tumor producing carpal tunnel syndrome and disabling pain. High‐resolution sonography allowed evaluation of gross tumor morphology and real‐time assessment of its interactions with surrounding structures. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2011
Soft tissue volume of upper arm in predicting small‐for‐gestational‐age fetuses using three‐dimensional ultrasound
Authors: Chang Chiung‐Hsin , Tsai Pei‐Yin , Yu Chen‐Hsiang , Ko Huei‐Chen , Chang Fong‐Ming
Published: October 2010

Purpose

To assess the value of fetal soft tissue volume (STV) of the upper arm in predicting small‐for‐gestational‐age (SGA) fetuses using three‐dimensional (3D) ultrasound (US).

Methods

We used 3D US to test the accuracy of fetal STV of the upper arm measurement in predicting SGA in a prospective cross‐sectional study.

Results

Fetal STV of the upper arm assessed by 3D US can differentiate SGA fetuses from appropriate‐for‐gestational‐age (AGA) fetuses. Using the 5th percentile as the cutoff, the sensitivity of fetal upper arm STV in predicting SGA fetuses was 84.1%, specificity, 93.4%, positive predictive value, 71.1%, negative predictive value, 96.8%, and overall accuracy, 91.9%. In addition, the diagnostic accuracy of fetal arm STV was better than that of the biparietal diameter, abdominal circumference, and femur length.

Conclusion

Fetal STV of upper arm assessment by 3D US is a novel method to predict SGA fetuses. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2011
Sonographic appearance of a cholesterol granuloma within a renal epithelioid angiomyolipoma
Authors: Cheng Kai‐Lun , Chang Han , Tsai Chung‐Ming , Tyan Yeu‐Sheng , Tsao Teng‐Fu
Published: October 2010

We present the sonographic characteristics of a rare case of cholesterol granuloma within a renal epithelioid angiomyolipoma (AML) in a 57‐year‐old woman. Sonography showed acoustic shadowing distal to the central region of the AML behind a hyperechoic cholesterol, which was hypoattenuating on CT with no contrast enhancement. Microscopic examination confirmed that this central area was a cholesterol granuloma. The patient was treated successfully with radical nephrectomy. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Automated image analysis for tracking cargo transport in axons
Authors: Zhang Kai , Osakada Yasuko , Xie Wenjun , Cui Bianxiao
Published: October 2010

The dynamics of cargo movement in axons encodes crucial information about the underlying regulatory mechanisms of the axonal transport process in neurons, a central problem in understanding many neurodegenerative diseases. Quantitative analysis of cargo dynamics in axons usually includes three steps: (1) acquiring time‐lapse image series, (2) localizing individual cargos at each time step, and (3) constructing dynamic trajectories for kinetic analysis. Currently, the later two steps are usually carried out with substantial human intervention. This article presents a method of automatic image analysis aiming for constructing cargo trajectories with higher data processing throughput, better spatial resolution, and minimal human intervention. The method is based on novel applications of several algorithms including 2D kymograph construction, seed points detection, trajectory curve tracing, back‐projection to extract spatial information, and position refining using a 2D Gaussian fitting. This method is sufficiently robust for usage on images with low signal‐to‐noise ratio, such as those from single molecule experiments. The method was experimentally validated by tracking the axonal transport of quantum dot and DiI fluorophore‐labeled vesicles in dorsal root ganglia neurons. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.
Analytic differential approach for robust registration of rat brain histological images
Authors: Hsu Wei‐Yen
Published: October 2010

Image registration is an important topic in medical image analysis. It is usually used to reconstruct 3D structure of tissues from a series of microscopic images. However, a variety of inherent factors may result in great differences between acquired slices during imaging even if they are adjacent. The common differences include the color difference and geometry discrepancy, which make the registration problem a difficult challenge. In this study, we propose a robust registration method to automatically reconstruct 3D volume data of the rat brain. It mainly consists of three procedures, including multiscale wavelet‐based feature extraction, analytic robust point matching (ARPM), and registration refinement with feature‐based modified Levenberg‐Marquardt algorithm (FMLM). The product of gradient moduli in multi‐scales is used to decide if extracted feature points are true according to the characteristic that features could exist in multiscale. The ARPM registration algorithm is proposed to speedily accomplish the registration of two point sets with different size by simultaneously evaluating the spatial correspondence and geometrical transformation. In addition, a FMLM method is also proposed to further refine registration results and achieve subpixel accuracy. The FMLM method converges much faster than most other methods due to its feature‐based and nonlinear characteristic. The performance of proposed method is evaluated by comparing it with well‐known thin‐plate spline robust point matching (TPS‐RPM) algorithm. The results indicate that ARPM‐FMLM algorithm is a robust and fast method in image registration. Microsc. Res. Tech., 2010. © 2010 Wiley‐Liss, Inc.
Histological and ultrastructural study of the intracellular behavior of indium in the testicular tissues
Authors: Samira Maghraoui , Ahlem Ayadi , Aouatef Ben Ammar , Habib Jaafoura Mohamed , Leila Tekaya
Published: October 2010

Indium, a IIIA group element of the periodic chart, has many medical uses for diagnostic and clinical investigations in humans. This element is also used in industry and in nuclear fields where released streams can contaminate environment. Consequently, indium can reach humans mainly by natural ways. In this work, we attempted to study the incidence of this element on the food intake and body and testicle weights of rat, as well as the histological and the ultrastructural consequences of its presence in testicles using conventional transmission electron microscopy. Our study showed that this element induced a significant decrease in the food intake and body and testicles weights and caused necrosis and vacuolization in germinal cells. The ultrastructural observations showed the presence of electron‐dense deposits characteristic of indium in the lysosomes of Leydig and Sertoli cells as well as sufferance in mitochondria of indium‐treated rats. Despite the role of lysosome in the protection of living cells, by sequestration and concentration of indium in testicle cells under insoluble form, it is probable that this element has noxious effects on food intake and body and testicles weight and induces necrosis on seminal tissues of treated rats. Microsc. Res. Tech., 2010. © 2010 Wiley‐Liss, Inc.
Immunohistochemical and immunochemical characterization of the distribution of leptin‐like proteins in the gastroenteric tract of two teleosts (Dicentrarchus labrax and Carassius auratus L.) with different feeding habits
Authors: Russo Finizia , De Girolamo Paolo , Neglia Simona , Gargiulo Annagiusi , Arcamone Nadia , Gargiulo Giuliana , Varricchio Ettore
Published: October 2010

Leptin is a modulator of food intake and energy homeostasis both in mammals and in some species of nonmammals vertebrates. In this study, we reported for the first time, using an immunohistochemical and immunochemical approach, the presence and distribution of immunoreactivity to leptin‐like protein in the gastroenteric tract of Dicentrarchus labrax (bass) and Carassius auratus (goldfish), two teleostean species with different feeding and different adaptative morphological organization of the gastroenteric tract. Bass stomach showed intense immunoreactivity to leptin‐like protein in all regions, with immunoreactive cells located at the base of the mucosal plicae and at the apical margin of the gastric crypts. Immunoreactive fibers and neuronal cells were observed close to vascular structures in the pyloric region. In bass and goldfish intestine, rare immunoreactive cells were observed along the mucosal epithelium mostly at the base or the apex of intestinal folds in the proximal and medium intestine; numerous immunoreactive nerve fibers in the circular and longitudinal layers of the tunica muscolaris as well as in the myenteric plexus were observed. Western blot analysis recognized a ∼15 kDa signal with a similar expression pattern for goldfish and sea bass. Our results could contribute to confirm the evolutive conservation of leptin‐like proteins and their probably precocious functional diversification in fish. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.
A simple and rapid dispersive liquid–liquid microextraction method followed by GC‐FID for determination ofN‐methylpyrrolidine in cefepime
Authors: Farajzadeh Mir Ali , Goushjuii Leila , Bashour Yousef
Published: October 2010

A simple, rapid and efficient sample preparation technique, dispersive liquid–liquid microextraction, coupled with gas chromatography‐flame ionization detection has been developed to determineN‐methylpyrrolidine in cefepime. The effect of various experimental factors on the preparation procedure, such as the nature and volume of extraction and disperser solvents, extraction time, the nature of buffer and its pH, and salt effect, was investigated, optimized and the following results were obtained: extraction solvent, chloroform; dispersive solvent and solvent for dissolving cefepime, a mixture of methanol/water (88:12, v/v); salting out agent, NaCl; and buffer, carbonate/bicarbonate (C=0.5 M, pH=12). The optimized conditions were applied to the real sample (cefepime) for the extraction and determination ofN‐methylpyrrolidine. The calibration graph is linear from 0.02 to 850 mg/L with the square of correlation coefficient 0.999. LOD and LOQ are 6.4 and 21.2 μg/L in solution, respectively, and 0.2 (2×10−5) and 0.6 (6×10−5) μg/g (%, w/w) in cefepime powder, respectively, using sample size 50 mg. Repeatability of the method is good and RSD% for six repeated experiments (C=170 mg/L) is 6.35%.
Single‐wall carbon nanotubes assisted photothermal cancer therapy: Animal study with a murine model of squamous cell carcinoma
Authors: Huang Naiyan , Wang Hequn , Zhao Jianhua , Lui Harvey , Korbelik Mladen , Zeng Haishan
Published: October 2010

Background and Objectives

There has been a dramatic increase in photothermal therapy as a minimally invasive treatment modality for cancer treatment due to the development of novel nanomaterials as the light absorption agents. Single‐wall carbon nanotubes (SWNTs) with strong optical absorption in the broad visible and near IR offer unique advantages for photothermal cancer therapy. A broad range of wavelengths can be used for the treatment with SWNTs, whereas conventional photothermal therapeutic agent is designed to absorb light only near one selected wavelength. The objective of this study is to validate the hypothesis that intratumoral injected SWNTs can absorb 785 nm near IR laser light and generate significant local hyperthermia to destroy tumors.

Study Design/Materials and Methods

SCCVII tumor in C3H/HeN mice was exposed to 785‐nm laser after intratumoral injection of SWNTs with different light and SWNTs dose combinations. The temperatures of the tumor with laser irradiation were monitored. In vivo and ex vivo Raman spectra in different organs were obtained with a rapid Raman system. Tumor responses (tumor volume and mouse survival) were documented daily after treatment up to day 45 to assess the effectiveness of the treatment.

Results

The temperature within the tumors increased in a light‐ and SWNTs‐dose dependent manner. Squamous cell carcinomas can be eradicated at a moderate light irradiance and fluence (200 mW/cm2 and 120 J/cm2). This light dose is also comparable to those used with photodynamic therapy. Tissue Raman spectroscopy measurements revealed that SWNTs remained localized in the tumor even 3 months after injection but was not found in other organs.

Conclusions

This animal study represents a significant step forward towards the goal of advancing SWNTs based photothermal cancer therapy into clinical applications. Lasers Surg. Med. 42:798–808, 2010 © 2010 Wiley‐Liss, Inc.
A Personalized Approach to Metabolic Aspects of Obesity
Authors: Brietzke Stephen A.
Published: October 2010

Metabolic syndrome, which is entwined in semantic controversy as to its actual existence as a distinct entity, links several important health conditions with obesity, and more specifically, excessive visceral adiposity. The most common linked disease states include type 2 diabetes mellitus, hypertension, dyslipidemia, obstructive sleep apnea, and cardiovascular and coronary heart disease. Much of the controversy surrounding the metabolic syndrome case definition is the purported centrality of insulin resistance as root cause, there being no universally agreed‐upon standard for measurement of insulin resistance. Over the past decade, the visceral adipocyte itself has emerged as a key contributor rather than passive bystander in the genesis of the metabolic syndrome. Rather than being a simple storage bin for excess triglyceride, the visceral adipocyte is an active endocrine cell secreting a variety of signal hormones known in the aggregate as adipokines. In optimal health, the predominant recognized adipokine is adiponectin, with downstream insulin‐sensitizing, anti‐inflammatory, antithrombotic, provasodilatory effects systemically. By contrast, metabolic syndrome is characterized by reduced adiponectin and increased inflammatory adipokine secretion, with downstream effects of insulin resistance, heightened inflammation, prothrombosis, and vasoconstriction. These alternative metabolic states of the adipocyte are characterized in this review as metabolic “yin” and “yang.” Lifestyle modifications and drug therapies that promote weight loss, increased physical exercise activity, and increased adiponectin production tend to modulate the system favorably toward metabolic “yin.” Mt Sinai J Med 77:499–510, 510, 2010. © 2010 Mount Sinai School of Medicine
Medical Therapy for Obesity
Authors: Bray George A.
Published: October 2010

Obesity results from a prolonged small positive energy imbalance, and treatment needs to reverse this imbalance. Many different diets have been tried to treat obesity, and weight loss occurs with all of them. There is currently no evidence that supports the superiority of one macronutrient composition for diets over any other. The principal effect seems to be the degree of adherence to the prescribed calorie reduction. Obesity drugs have been developed that tap brain mechanisms for controlling feeding and the gastrointestinal tract and its peptides. Orlistat blocks intestinal lipase and produces modest weight loss. Sibutramine is a serotonin‐norepinephrine reuptake inhibitor that has a warning on its label from the US Food and Drug Administration because of cardiovascular risk. Its marketing has been suspended in Europe. Several drug combinations are on the horizon for treatment of obesity.Mt Sinai J Med 77:407–417, 2010.. © 2010 Mount Sinai School of Medicine
Weight Management in Type 2 Diabetes Mellitus
Authors: Siram Amulya T. , Yanagisawa Robert , Skamagas Maria
Published: October 2010

Obesity is a well known risk factor for type 2 diabetes mellitus. Individuals with type 2 diabetes mellitus are at risk for weight gain as a result of multiple influences, including sedentary lifestyle, high‐calorie diet, diabetes medications, sociocultural factors, chronic medical and psychiatric illnesses, and a dysregulated enteroendocrine axis. Because both diabetes mellitus and obesity predispose patients to abnormal cardiometabolic profiles and increased cardiovascular disease, management of diabetes mellitus should focus on weight management and optimizing cardiometabolic parameters, concomitant with glycemic control. Lifestyle modification incorporating healthy, calorie‐appropriate diets and increased physical activity, in addition to metformin, are central components to diabetes management and weight management. These interventions have been shown to improve body weight, glycemic control, and overall cardiometabolic profile. The weight‐neutral and weight‐losing diabetes medications include metformin, alpha‐glucosidase inhibitors, glucagon‐like peptide‐1 analogs, dipeptidyl peptidase‐4 inhibitors, and amylin analogs. It is essential that providers understand the metabolic and weight effects of diabetes medications in order to develop strategies for managing diabetes mellitus while helping patients maintain or lose weight in order to improve their overall health outcomes. Mt Sinai J Med 77:533–548,2010. © 2010 Mount Sinai School of Medicine
miRNAs in human cancer
Authors: Farazi Thalia A , Spitzer Jessica I , Morozov Pavel , Tuschl Thomas
Published: October 2010

Mature microRNAs (miRNAs) are single‐stranded RNA molecules of 20–23 nucleotide (nt) length that control gene expression in many cellular processes. These molecules typically reduce the stability of mRNAs, including those of genes that mediate processes in tumorigenesis, such as inflammation, cell cycle regulation, stress response, differentiation, apoptosis and invasion. miRNA targeting is mostly achieved through specific base‐pairing interactions between the 5′ end (‘seed’ region) of the miRNA and sites within coding and untranslated regions (UTRs) of mRNAs; target sites in the 3′ UTR lead to more effective mRNA destabilization. Since miRNAs frequently target hundreds of mRNAs, miRNA regulatory pathways are complex. To provide a critical overview of miRNA dysregulation in cancer, we first discuss the methods currently available for studying the role of miRNAs in cancer and then review miRNA genomic organization, biogenesis and mechanism of target recognition, examining how these processes are altered in tumorigenesis. Given the critical role miRNAs play in tumorigenesis processes and their disease‐specific expression, they hold potential as therapeutic targets and novel biomarkers. Copyright © 2010 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Metabolomic pattern of childhood neuroblastoma obtained by 1H‐high‐resolution magic angle spinning (HRMAS) NMR spectroscopy
Authors: Imperiale Alessio , Elbayed Karim , Moussallieh François‐Marie , Neuville Agnès , Piotto Martial , Bellocq Jean‐Pierre , Lutz Patrick , Namer Izzie‐Jacques
Published: October 2010

Background

The aim of this preliminary study is to characterize by 1H high‐resolution magic angle spinning NMR spectroscopy (HRMAS) the metabolic content of intact biopsy samples obtained from 12 patients suffering from neuroblastoma (NB).

Procedure

The biochemical NB profile was first compared to normal adrenal medulla. In a second step, the relationship between the tumor metabolic profile and the patients' clinical data was investigated.

Results

A higher level of creatine, glutamine/glutamate, acetate and glycine characterized NB biopsies while healthy adrenal medulla tissue contained adrenaline and a larger amount of ascorbic acid. Adrenaline, which was undetectable in NB spectra, represented the metabolic signature of normal adrenal medulla. NB from patients younger than 12 months contained a higher level of acetate and lysine. Conversely, higher amounts of glutathione, glutamate, myo‐inositol, glycine, serine and ascorbic acid were detected in NB samples belonging to younger children. Glutamine/glutamate, aspartate, creatine, glycine were characteristic of stage I–II NB. Acetate and creatine were characteristic of stage IV NB. Finally, a relatively higher amount of aspartate, succinate, and glutathione was detected in patients alive without active disease after a mean follow‐up of 7 years whereas a higher concentration of acetate and taurine was characteristic of patients with worse prognosis.

Conclusions

Our preliminary results suggest the existence of a complex metabolic reality in NB, probably representative of tumor behavior. However, the real impact of these promising results should be assessed by long‐term prospective studies on a larger cohort of patients. Pediatr Blood Cancer. 2010;56:24–34. © 2010 Wiley‐Liss, Inc.
Pyuria is absent during urinary tract infections in neutropenic patients
Authors: Klaassen I.L.M. , de Haas V. , van Wijk J.A.E. , Kaspers G.J.L. , Bijlsma M. , Bökenkamp A.
Published: October 2010

To investigate the diagnostic significance of a normal urine sediment in the work‐up for fever of unknown origin in neutropenia. Urinary tract infection was defined as ≥105 urinary pathogens in the absence of another focus. Pyuria was found in only 1/23 neutropenic episodes compared to 21/31 in controls (P < 0.0001). Pediatr Blood Cancer 2011;56:868–870. © 2010 Wiley‐Liss, Inc.
Proteome and radioimmunoassay analyses of pituitary hormones and proteins in response to feed restriction of dairy cows
Authors: Kuhla Björn , Albrecht Dirk , Bruckmaier Rupert , Viergutz Torsten , Nürnberg Gerd , Metges Cornelia C.
Published: October 2010

The hypothalamic‐pituitary system controls homeostasis during feed energy reduction. In order to examine which pituitary proteins and hormone variants are potentially associated with metabolic adaptation, pituitary glands from ad libitum and energy restrictively fed dairy cows were characterized using RIA and 2‐DE followed by MALDI‐TOF‐MS. We found 64 different spots of regulatory hormones: growth hormone (44), preprolactin (16), luteinizing hormone (LH) (1), thyrotropin (1), proopiomelanocortin (1) and its cleavage product lipotropin (1), but none of these did significantly differ between feeding groups. Quantification of total pituitary LH and prolactin concentrations by RIA confirmed the results obtained by proteome analysis. Also, feed energy restriction provoked increasing non‐esterified fatty acid, decreasing prolactin, but unaltered glucose, LH and growth hormone plasma concentrations. Energy restriction decreased the expression of glial fibrillary acidic protein, triosephosphate isomerase, purine‐rich element‐binding protein A and elongation factor Tu, whereas it increased expression of proline synthetase co‐transcribed homolog, peroxiredoxin III, β‐tubulin and annexin A5 which is involved in the hormone secretion process. Our results indicate that in response to feed energy restriction the pituitary reservoir of all posttranslationally modified hormone forms remains constant. Changing plasma hormone concentrations are likely attributed to a regulated releasing process from the gland into the blood.
Impact of oxidation on protein therapeutics: Conformational dynamics of intact and oxidized acid‐β‐glucocerebrosidase at near‐physiological pH
Authors: Bobst Cedric E. , Thomas John J. , Salinas Paul A. , Savickas Philip , Kaltashov Igor A.
Published: October 2010

The solution dynamics of an enzyme acid‐β‐glucocerebrosidase (GCase) probed at a physiologically relevant (lysosomal) pH by hydrogen/deuterium exchange mass spectrometry (HDX‐MS) reveals very uneven distribution of backbone amide protection across the polypeptide chain. Highly mobile segments are observed even within the catalytic cavity alongside highly protective segments, highlighting the importance of the balance between conformational stability and flexibility for enzymatic activity. Forced oxidation of GCase that resulted in a 40–60% reduction in in vitro biological activity affects the stability of some key structural elements within the catalytic site. These changes in dynamics occur on a longer time scale that is irrelevant for catalysis, effectively ruling out loss of structure in the catalytic site as a major factor contributing to the reduction of the catalytic activity. Oxidation also leads to noticeable destabilization of conformation in remote protein segments on a much larger scale, which is likely to increase the aggregation propensity of GCase and affect its bioavailability. Therefore, it appears that oxidation exerts its negative impact on the biological activity of GCase indirectly, primarily through accelerated aggregation and impaired trafficking.
TEV protease‐facilitated stoichiometric delivery of multiple genes using a single expression vector
Authors: Chen Xi , Pham Elizabeth , Truong Kevin
Published: October 2010

Delivery and expression of multiple genes is an important requirement in a range of applications such as the engineering of synthetic signaling pathways and the induction of pluripotent stem cells. However, conventional approaches are often inefficient, nonstoichiometric and may limit the maximum number of genes that can be simultaneously expressed. We here describe a versatile approach for multiple gene delivery using a single expression vector by mimicking the protein expression strategy of RNA viruses. This was accomplished by first expressing the genes together with TEV protease as a single fusion protein, then proteolytically self‐cleaving the fusion protein into functional components. To demonstrate this method in E. coli cells, we analyzed the translation products using SDS‐PAGE and showed that the fusion protein was efficiently cleaved into its components, which can then be purified individually or as a binding complex. To demonstrate this method in mammalian cells, we designed a differential localization scheme and used live cell imaging to observe the distinctive subcellular targeting of the processed products. We also showed that the stoichiometry of the processed products was consistent and corresponded with the frequency of appearance of their genes on the expression vector. In summary, the efficient expression and separation of up to three genes was achieved in both E. coli and mammalian cells using a single TEV protease self‐processing vector.
Preparation of a highly translocation‐competent proOmpA/SecB complex
Authors: Nishiyama Ken‐Ichi , Tokuda Hajime
Published: October 2010

Methods for reproducibly preparing highly translocation‐competent proOmpA were developed. Only a competent form of proOmpA was sorted out from incompetent one using SecB, a translocation‐dedicated chaperone, as a probe. Trypsin digestion revealed that the incompetent form of proOmpA was partially folded at its N‐terminus, consistent with the jamming of proOmpA within translocon. Although the incompetent form of proOmpA was not active as to topology inversion of SecG, the isolated proOmpA/SecB complex had recovered the ability of SecG inversion. These results let us prepare a proOmpA/SecB complex both in vivo and in vitro that is highly translocation‐competent. E. coli cells harboring a plasmid, in which ompA and secB were encoded as a synthetic operon, accumulated the proOmpA/SecB complex in the cytosol. The complex, purified by means of a His tag attached to SecB, was found to be translocation‐competent as revealed by the occurrence of SecG inversion, although the signal peptide of proOmpA was sensitive to proteolytic digestion. ProOmpA, in vitro synthesized by means of a continuous exchange cell free system in the presence of SecB‐His, was purified as a complex with SecB, which was active as to SecG inversion as well.
Evidence of epithelial to mesenchymal transition associated with increased tumorigenic potential in an immortalized normal prostate epithelial cell line
Authors: Marian Calin O. , Yang Lin , Zou Ying S. , Gore Crystal , Pong Rey‐Chen , Shay Jerry W. , Kabbani Wareef , Hsieh Jer‐Tsong , Raj Ganesh V.
Published: October 2010

BACKGROUND

The majority of established human prostate cancer cell lines are derived from metastatic lesions and are already tumorigenic in vivo, therefore immortalized normal prostate cell lines may provide a more relevant model to unveil the mechanisms associated with cancer progression and metastasis.

METHODS

PZ‐HPV‐7, an immortalized human prostate epithelial cell line was used to generate xenograft tumors in mice. A subline designated HPV‐PZ‐7T was subsequently derived from the subrenal capsule xenograft of a nude mouse. These cells were further characterized using karyotyping, immunofluorescence, qRT‐PCR, Western blotting, and three‐dimensional cultures in Matrigel.

RESULTS

The PZ‐HPV‐7 cell line possesses a typical epithelial morphology, expresses basal cell markers, and is capable of forming web‐like structures with evidence of budding on Matrigel. PZ‐HPV‐7 is non‐tumorigenic in immunocompromised mice by either subcutaneous injection or subrenal grafting. In contrast, the PZ‐HPV‐7T cells, derived from a xenograft tumor induced by co‐inoculation with matrigel using subrenal grafting, possess a mesenchymal phenotype as well as luminal cell markers and are highly tumorigenic and metastatic in nude mice. Functionally and biochemically, the PZ‐HPV‐7T subline appears to have undergone an epithelial‐to‐mesenchymal transition (EMT) from the parental PZ‐HPV‐7 line.

CONCLUSION

We have developed a novel EMT model using an immortalized normal prostate epithelial cell line and generated a new prostate cancer cell line, PZ‐HPV‐7T, which may represent an excellent system to study mechanisms associated with prostate cancer progression and metastasis. Prostate 71:626–636, 2011. © 2010 Wiley‐Liss, Inc.
Gastrin‐releasing peptide: Predictor of castration‐resistant prostate cancer?
Authors: Heinrich Elmar , Probst Kai , Michel Maurice Stephan , Trojan Lutz
Published: October 2010

BACKGROUND

Neuroendocrine (NE) cells of the prostate are known to be androgen‐independent and NE peptides like gastrin‐releasing peptide (GRP) or neuron‐specific enolase (NSE) can stimulate growth in a paracrine manner, and this is thought to be one of the escape mechanisms in castration‐resistant prostate cancer (CRPCa). In a longitudinal study, we investigated the development of the NE serum factors GRP, NSE, and chromogranin A and their correlation with prostate‐specific androgen (PSA) during hormonal treatment.

MATERIALS AND METHODS

Thirty two patients, with histology‐proven, localized or metastatic prostatic carcinoma (PCa), who were undergoing therapy with LHRH analogue or a combination of LHRH analog and peripheral androgen blockade, took part in the study. In addition, eight healthy volunteers were each tested twice for serum GRP to elicit a “physiological” standard value. Blood samples were taken periodically from each patient within an 18‐month time frame.

RESULTS

We defined the standard value for GRP in the healthy participants as 0.852 ng/ml (mean + 2 SD) and observed that the GRP values for patients with PCa were significantly higher (P = 0.034). There was a positive correlation between PSA and GRP in patients with biochemical failure. CgA correlated with PSA development in the CRPCa patients. NSE values rose steadily over the study period, but with no correlation to PSA.

CONCLUSION

Our data confirm that NE factors are elevated during hormonal treatment of prostate cancer. GRP is higher in PCa patients undergoing androgen deprivation therapy and is possibly involved in the initiation of hormonal escape in PCa. Prostate 71:642–648, 2011. © 2010 Wiley‐Liss, Inc.
LHRH antagonist Cetrorelix reduces prostate size and gene expression of proinflammatory cytokines and growth factors in a rat model of benign prostatic hyperplasia
Authors: Rick Ferenc G. , Schally Andrew V. , Block Norman L. , Halmos Gabor , Perez Roberto , Fernandez Jesus B. , Vidaurre Irving , Szalontay Luca
Published: October 2010

BACKGROUND

Recent findings suggest that BPH has an inflammatory component. Clinical trials have documented that therapy with LHRH antagonist Cetrorelix causes a marked and prolonged improvement in LUTS in men with symptomatic BPH. We investigated the mechanism of action and effect of Cetrorelix in a rat model of BPH.

METHODS

Adult male Wistar rats were used. BPH was induced in rats by subcutaneous injections of TE 2 mg/day for 4 weeks. Control animals received injections of corn oil. After induction of BPH, rats received depot Cetrorelix pamoate at the doses of 0.625, 1.25, and 12.5 mg/kg on days 1 and 22 and TE‐control rats received vehicle injections. Whole prostates were weighed and processed for RNA and protein. Real‐time RT‐PCR assays for numerous inflammatory cytokines and growth factors were performed. Quantitative analyses of prostatic LHRH receptor, LHRH, androgen receptor (AR) and 5α‐reductase 2 were done by real‐time RT‐PCR and immunoblotting; serum DHT, LH, PSA, and IGF‐1 by immunoassays.

RESULTS

mRNA levels for inflammatory cytokines IFN‐γ, IL‐3, IL‐4, IL‐5, IL‐6, IL‐8, IL‐13, IL‐15, and IL‐17 and for growth factors EGF, FGF‐2, FGF‐7, FGF‐8, FGF‐14, TGF‐β1, and VEGF‐A were significantly reduced by Cetrorelix 0.625 mg/kg (P < 0.05). Prostate weights were also significantly lowered by any dose of Cetrorelix.

CONCLUSIONS

This study suggests that Cetrorelix reduces various inflammatory cytokines and growth factors in rat prostate and, at doses which do not induce castration levels of testosterone, can lower prostate weights. Our findings shed light on the mechanism of action of LHRH antagonists in BPH. Prostate 71:736–747, 2011. © 2010 Wiley‐Liss, Inc.
Immunoreactivity of the salivary protein statherin in human male accessory sex glands
Authors: Cossu Margherita , Isola Michela , Solinas Paola , De Lisa Antonello , Massa Denise , Lantini Maria S.
Published: October 2010

BACKGROUND

Statherin is a small phosphoprotein chiefly studied for its protective roles towards teeth and oral tissues. Although generally considered as exclusively secreted by salivary glands, circumstantial evidences suggested that other tissues also produce it. This article first demonstrates statherin immunoreactivity in human prostate and seminal vesicles.

METHODS

Surgical samples of prostate and seminal vesicles were fixed in a mixture of paraformaldehyde and glutaraldehyde, and embedded in Epon resin without previous osmication. Ultrathin sections were treated for the intracellular localization of statherin by means of an immunogold staining method.

RESULTS

Reactive statherin was revealed in secreting cells of both seminal vesicle and prostate epithelia: labeling was found in secretory granules of seminal vesicle cells and in cytoplasmic vesicles of prostatic cells.

CONCLUSIONS

The different staining patterns suggested that the two glands secrete statherin through different pathways. Prostate 71:671–674, 2011. © 2010 Wiley‐Liss, Inc.
Multiple mechanisms account for resistance to sterol 14α‐demethylation inhibitors in field isolates of Mycosphaerella graminicola
Authors: Leroux Pierre , Walker Anne‐Sophie
Published: October 2010

BACKGROUND: Sterol 14α‐demethylation inhibitors (DMIs) have been widely used in many European countries to control septoria leaf blotch, which is caused by Mycosphaerella graminicola (Fückel) J Schrot (anamorph Septoria tritici Berk & MA Curtis). However, treatment efficacy has declined, and significant shifts in population susceptibility have occurred in recent years, with the isolation of particularly highly resistant strains from French, English and Irish populations. The present aim was to determine the phenotypic characteristics of these field isolates and to identify the possible resistance mechanisms.

RESULTS: Target alteration, linked to 11 possible changes in the gene encoding 14α‐demethylase (Cyp51), was the basic resistance mechanism in weakly, moderately and highly resistant strains. Changes in Cyp51 combined with the overexpression of drug efflux transporters probably result in multidrug resistance in some of the most resistant phenotypes. Finally, some moderately or highly resistant isolates were found to harbour an insertion in the Cyp51 promoter and/or new combinations of known mutations in the target gene.

CONCLUSION: An updated overview of M. graminicola field strains displaying low to high resistance to DMIs is provided here. The management of field resistance and efficacy should be adapted to take these findings into account. Copyright © 2010 Society of Chemical Industry