Effectiveness of a LED flashlight technique in reducing livestock depredation by lions (Panthera leo) around Nairobi National Park, Kenya.
The global lion (Panthera leo) population decline is partly a result of retaliatory killing in response to livestock depredation. Nairobi National Park (NNP) is a small protected area in Kenya surrounded by a human-dominated landscape. Communities around the park use flashlights to deter lions from their livestock bomas. We investigated the response by lions to the installation of a LED flashlight technique during 2007-2016.We interviewed 80 owners of livestock bomas with flashlights (n = 43) and without (n = 37) flashlights in the surroundings of NNP and verified reported attacks on bomas against predation data over10 years. The frequency of attacks on bomas equipped with flashlights was significantly lower compared to bomas without flashlights. We also found that after flashlight installation at livestock bomas, lion attacks took place further away from the park edge, towards areas where bomas without flashlights were still present. With increased numbers of flashlight installations at bomas in recent years, we further noticed a shift from nocturnal to more diurnal predation incidences. Our study shows that the LED flashlight technique is effective in reducing nocturnal livestock predation at bomas by lions. Long term studies on the effects as well as expansion of this technique into other communities around NNP are recommended.


NETWORKED 3B: a novel protein in the actin cytoskeleton-endoplasmic reticulum interaction.
In plants movement of the endoplasmic reticulum (ER) is dependent on the actin cytoskeleton. However little is known about proteins that link the ER membrane and the actin cytoskeleton. Here we identified a novel protein, NETWORKED 3B (NET3B), which is associated with the ER and actin cytoskeleton in vivo. NET3B belongs to a superfamily of plant specific actin binding proteins, the NETWORKED family. NET3B associates with the actin cytoskeleton in vivo through an N-terminal NET actin binding (NAB) domain, which has been well-characterized in other members of the NET family. A three amino acid insertion, Val-Glu-Asp, in the NAB domain of NET3B appears to lower its ability to localize to the actin cytoskeleton compared with NET1A, the founding member of the NET family. The C-terminal domain of NET3B links the protein to the ER. Overexpression of NET3B enhanced the association between the ER and the actin cytoskeleton, and the extent of this association was dependent on the amount of NET3B available. Another effect of NET3B overexpression was a reduction in ER membrane diffusion. In conclusion, our results revealed that NET3B modulates ER and actin cytoskeleton interactions in higher plants.


Endoluminal minirobots for transgastric peritoneoscopy.
The performance of surgeries through small incisions or natural orifices minimizes the invasiveness to the patient as compared to open procedures. However, the constraints on visual feedback and dexterity limit the scope of these procedures. Recent robotic technologies attempt to mitigate these constraints for flexible endoscopy and laparoscopy. Much of the current work in flexible endoscopy is in the development of a fully autonomous endoscope capable of providing the surgeon with better control. Advancements in laparoscopic technologies have demonstrated abilities to improve visualization and dexterity through telerobotics and in vivo robotics. The application of new robotic technologies in flexible endoscopy and laparoscopy to natural orifice surgery promises to further benefit the patient by eliminating abdominal incisions, scarring, and the pain associated with these incisions. The safety and feasibility of such robotic technology has been successfully demonstrated for natural orifice transluminal endoscopic surgery (NOTES) in animal models.


Instabilities in a growing system of active particles: scalar and vectorial systems.
The physics of micron-scale biological colonies usually benefits from different out-of-equilibrium sources. In bacterial colonies and cellular tissues, the growth process is among the important active sources that determine the dynamics. In this article, we study the generic dynamical instabilities associated with the growth phenomena that may arise in both scalar and vectorial systems. In vectorial systems, where the rotational degrees of particles play a role, a phenomenological growth-mediated torque can affect the rotational dynamics of individual particles. We show that such a growth-mediated torque can result in active traveling waves in the bulk of a growing system. In addition to the bulk properties, we analyze the instabilities in the shape of growing interfaces in both scalar and vectorial systems.


[Two cases of long-term survival after treatment of recurrent intrahepatic cholangiocarcinoma and intrahepatic metastasis in the remnant liver with repeated hepatectomy].
We report 2 cases of recurrent intrahepatic cholangiocarcinoma (ICC) with intrahepatic metastasis in the remnant liver in patients who underwent repeated hepatectomy and are currently alive and free of the disease. Case 1 involves a 53-year- old woman who underwent extended right hepatectomy for mass-forming type ICC (7-cm in diameter) in the right lobe. After 7 months, she experienced tumor recurrence, and exhibited a 3.8-cm solitary intrahepatic metastasis in the remnant left lateral segment. The patient underwent partial resection of the lateral segment and is currently disease-free at 2 years and 9 months after the recurrence. Case 2 involves a 59-year-old woman who underwent extended left hepatectomy for combined mass-forming and periductal-infiltrating-type ICC( 5-cm in diameter) in the left lobe. After 2 years and 3 months, she experienced tumor recurrence, and exhibited a 3-cm solitary intrahepatic metastasis in the remnant right anterior segment 8. She underwent partial resection of segment 8 and is currently disease-free at 2 years and 11 months after the recurrence. Most of the recurrences of ICC in the remnant liver exhibit multiple lesions or concomitant extrahepatic metastases; therefore, the indication of repeated hepatectomy for intrahepatic recurrence in the remnant liver is restrictive. However, most of the reported cases on repeated hepatectomy for intrahepatic recurrence involved a solitary metastasis in the remnant liver, and revealed a relatively favorable prognosis. Thus, we believe that a solitary recurrence of ICC in the remnant liver could be an indication for repeated hepatectomy.


Analysis of urine cytology at a community hospital.
To review the pattern of urine cytology in our setting of a community hospital with age and gender distribution. Also to discuss the importance of this safe and inexpensive tool for diagnosing and screening urothelial cancers. We desire to encourage more wider and confident use of urine cytology in the routine practice. We reviewed all the consecutive urine cytology specimens received at our institution during time frame of one year. The cytological diagnosis were grouped in four categories; 1: negative, benign or reactive cases; 2: cases showing acute inflammatory changes; 3: inconclusive cases and 4: malignant cases. Only the positive malignant cases were followed histologically to determine the accuracy and efficiency of urine cytological examination for positive cases. A total of 1957 consecutive urine cytology specimens were reviewed. Majority of the cases were negative for tumour (67.19%). The most common diagnostic category was acute inflammatory case (17.3%) while inconclusive cases constituted 11.39% of total. There were 23 (1.18%) positive cases that were diagnosed malignant on urine cytology. These 23 malignant cases were followed histologically and 22 (95.6%) cases turned out to be malignant. Urine cytological examination is an efficient tool that has a good diagnostic yield in detecting malignant urothelial lesions. It should be used routinely for diagnosing, screening and for follow-up of all malignant urothelial lesions.


Functionalized cyclic RGD peptidomimetics: conjugable ligands for αvβ3 receptor imaging.
The α(v)β(3) integrin is an adhesion molecule involved in physiological and pathological angiogenesis as well as in tumor invasion and metastasis, and therefore, there is a strong interest in developing novel agents interacting with this molecule. We report the synthesis and characterization of fluorescent α(v)β(3) integrin probes and their use to visualize integrin α(v)β(3) expression on human normal and cancer cells. The fluorescent probes we describe here may be of use for noninvasive imaging of α(V)β(3) integrin expression also in vivo.


Electrocochleography in children. A retrospective study.
A follow-up study of 35 children subjected to electrocochleography (Ecog) was conducted to determine (1) agreement of Ecog results with other audiometric measures obtained subsequently, and (2) accuracy of recommendations for patient management made on the basis of Ecog results. The latter was determined by parental experience, educational placement and achievement, and information from medical and paramedical personnel currently managing the children. These data indicate that Ecog test results are valuable in helping select between possible management alternatives in children who are not testable by behavioral means. However, discussion of two "problem" cases points to possible errors of management that could be made if Ecog results were the only available diagnostic information.


[GBV-C infection in HIV-infected patients in the Russian Federation].
The spread and genotypical variability of GBV-C virus were determined among the HIV-positive patients in the Russian Federation. More than a fourth (26.2%) of the HIV-infected patients were shown to have GBV-C coinfection; all virus isolates belonged to genotype 2 with a predominance of subtype 2a. Analysis of the impact of GBV-C coinfection on HIV burden and CD4 lymphocyte levels showed no significant impact on these basic characteristics of HIV infection. However, coinfection with GBV-C and HIV was associated with the higher frequency of undetectably low ( < 400 copies/ml) of HIV burden, which enables GBV-C infection to be regarded as a potentially favorable factor in HIV infection.


[Repair of xiangsha liujunzi decoction on damage of interstitial cells of cajal and gap junction in the gastric muscular layer of rats of Pi-Qi deficiency syndrome: an experiment study].
To explore the repair of Xiangsha Liujunzi Decoction (XSLJZD) on interstitial cells of Cajal (ICC) and gap junction (GJ) in the gastric muscular layer of rats of Pi-qi deficiency syn- drome (PQDS). PQDS was established using purgative method with bitter and cold drugs in 30 healthy Wistar rats. After successful modeling they were randomly divided into the treatment group and the model group, 15 in each group. Another 15 healthy Wistar rats were recruited as the healthy control group. Rats in the treatment group were gastric administered with XSLJZD at 2 mL/100 g body weight, once daily for 14 successive days. Equal volume of normal saline was gastrically administered to those in the healthy control group and the model group. The gastric muscle tissues were taken out before modeling, before intervention, and after intervention, respectively. Ultrastructural changes of ICC and GJ were observed using transmission electron microscope (TEM). The number and distribution of Connexin43 (Cx43) were detected using immunohistochemistry. Results of TEM indicated that compared with the healthy control group, both ICC and GJ in the model group showed obvious injury. ICC and GJ were apparently repaired after intervention in the treatment group. Compared with the same group before modeling, the integrated optical density (IOD) of the Cx43 expression significantly decreased in the model group before and after intervention (P <0.05). Compared with before intervention, the IOD of the Cx43 expression significantly increased in the treatment group (P <0.05). Compared with the healthy control group, the IOD of the Cx43 expression significantly decreased in the model group before and after intervention (P <0.05). Compared with the model group, the IOD of the Cx43 expression significantly increased in the treatment group (P <0.05). Ultrastructures of ICC and GJ in the gastric muscular layer of rats of PQDS were obviously damaged. XSLJZD could repair the structural damage of ICC and GJ in the gastric muscle tissues of rats of PQDS.


High-dose oral dextromethorphan versus placebo in painful diabetic neuropathy and postherpetic neuralgia.
N-methyl-D-aspartate (NMDA) receptor antagonists relieve neuropathic pain in animal models, but side effects of dissociative anesthetic channel blockers, such as ketamine, have discouraged clinical application. Based on the hypothesis that low-affinity NMDA channel blockers might have a better therapeutic ratio, we carried out two randomized, double-blind, crossover trials comparing six weeks of oral dextromethorphan to placebo in two groups, made up of 14 patients with painful distal symmetrical diabetic neuropathy and 18 with postherpetic neuralgia. Thirteen patients with each diagnosis completed the comparison. Dosage was titrated in each patient to the highest level reached without disrupting normal activities; mean doses were 381 mg/day in diabetics and 439 mg/day in postherpetic neuralgia patients. In diabetic neuropathy, dextromethorphan decreased pain by a mean of 24% (95% CI: 6% to 42%, p = 0.01), relative to placebo. In postherpetic neuralgia, dextromethorphan did not reduce pain (95% CI: 10% decrease in pain to 14% increase in pain, p = 0.72). Five of 31 patients who took dextromethorphan dropped out due to sedation or ataxia during dose escalation, but the remaining patients all reached a reasonably well-tolerated maintenance dose. We conclude that dextromethorphan or other low-affinity NMDA channel blockers may have promise in the treatment of painful diabetic neuropathy.


The movement protein of cucumber mosaic virus traffics into sieve elements in minor veins of nicotiana clevelandii
The location of the 3a movement protein (MP) of cucumber mosaic virus (CMV) was studied by quantitative immunogold labeling of the wild-type 3a MP in leaves of Nicotiana clevelandii infected by CMV as well as by using a 3a-green fluorescent protein (GFP) fusion expressed from a potato virus X (PVX) vector. Whether expressed from CMV or PVX, the 3a MP targeted plasmodesmata and accumulated in the central cavity of the pore. Within minor veins, the most extensively labeled plasmodesmata were those connecting sieve elements and companion cells. In addition to targeting plasmodesmata, the 3a MP accumulated in the parietal layer of mature sieve elements. Confocal imaging of cells expressing the 3a-GFP fusion protein showed that the 3a MP assembled into elaborate fibrillar formations in the sieve element parietal layer. The ability of 3a-GFP, expressed from PVX rather than CMV, to enter sieve elements demonstrates that neither the CMV RNA nor the CMV coat protein is required for trafficking of the 3a MP into sieve elements. CMV virions were not detected in plasmodesmata from CMV-infected tissue, although large CMV aggregates were often found in the parietal layer of sieve elements and were usually surrounded by 3a MP. These data suggest that CMV traffics into minor vein sieve elements as a ribonucleoprotein complex that contains the viral RNA, coat protein, and 3a MP, with subsequent viral assembly occurring in the sieve element parietal layer.


Audiogenic seizure protection by elevated brain GABA concentration in mice: effects of gamma-acetylenic gaba and gamma-vinyl GABA, two irreversible GABA-T inhibitors.
gamma-Acetylenic GABA and gamma-vinyl GABA, two catalytic irreversible inhibitors of GABA-transaminase, produce marked and sustained elevations in mouse brain GABA concentrations and protect DBA/2 mice against audiogenically induced seizures in a similar dose and time-dependent manner. The acetylenic analog also inhibits GAD activity while the vinyl compound has minimal activity against this enzyme. The increase in brain GABA concentrations induced by these compounds correlates well with attenuation of audiogenic seizure intensity (r = 0.991 and 0.962 for gamma-acetylenic and gamma-vinyl GABA respectively) and with degree of seizure protection (r = 0.974 and 0.834). Seizure intensity is reduced by 50% when brain GABA is increased to 265% and 264% of control values by the two inhibitors and seizure incidence is halved at 322% and 324%. Thus, audiogenic seizure protection in genetically susceptible mice is apparently a function of whole brain GABA concentrations.


Polymorphisms in the HBB gene relate to individual cardiorespiratory adaptation in response to endurance training.
The crucial role of haemoglobin in endurance performance has been well documented. We examined whether polymorphisms in the HBB gene modified aerobic capacity. 102 recruits were trained by running 5000 m three times per week for 18 weeks. Exercise intensity progressively increased from an initial heart rate corresponding to 95% of the individual baseline ventilatory threshold during the first 10 weeks to 105% during the last 8 weeks. The phenotypes measured were running economy and VO(2)max. Running economy was determined by measuring submaximal VO(2) for 5 min at a constant running speed of 12 km.h(-1) and VO(2)max was obtained during an incremental test to exhaustion. Genomic DNA was extracted from white cells of peripheral blood and the -551C/T, intron2,+16C/G and +340 A/T genotypes were examined relative to the TAA site variants by PCR-RFLP. Genotype distributions were in Hardy-Weinberg equilibrium at three loci. None of the running economy and VO(2)max-related traits were associated with the three polymorphisms or haplotypes at baseline, while the training response of running economy was associated with -551C/T and intron2,+16C/G polymorphisms. Subjects homozygous for intron2,+16C/C or -551C/C had decreased oxygen cost of running compared to the other individuals. It was concluded that the -551C/C or intron2,+16C/C genotype might explain part of the individual variation in the cardiorespiratory adaptation to endurance training.


Precision vs flexibility in GPCR signaling.
The G protein coupled receptor (GPCR) rhodopsin activates the heterotrimeric G protein transducin (Gt) to transmit the light signal into retinal rod cells. The rhodopsin activity is virtually zero in the dark and jumps by more than one billion fold after photon capture. Such perfect switching implies both high fidelity and speed of rhodopsin/Gt coupling. We employed Fourier transform infrared (FTIR) spectroscopy and supporting all-atom molecular dynamics (MD) simulations to study the conformational diversity of rhodopsin in membrane environment and extend the static picture provided by the available crystal structures. The FTIR results show how the equilibria of inactive and active protein states of the receptor (so-called metarhodopsin states) are regulated by the highly conserved E(D)RY and Yx7K(R) motives. The MD data identify an intrinsically unstructured cytoplasmic loop region connecting transmembrane helices 5 and 6 (CL3) and show how each protein state is split into conformational substates. The C-termini of the Gtγ- and Gtα-subunits (GαCT and GγCT), prepared as synthetic peptides, are likely to bind sequentially and at different sites of the active receptor. The peptides have different effects on the receptor conformation. While GγCT stabilizes the active states but preserves CL3 flexibility, GαCT selectively stabilizes a single conformational substate with largely helical CL3, as it is found in crystal structures. Based on these results we propose a mechanism for the fast and precise signal transfer from rhodopsin to Gt, which assumes a stepwise and mutual reduction of their conformational space. The mechanism relies on conserved amino acids and may therefore underlie GPCR/G protein coupling in general.


[The significance of the Balint-group work: a review of the literature and an empirical study in Unterfranken].
The group work initiated by Michael Balint aims at a comprehensive understanding of the doctor-patient-relationship and a patient-centered medicine. In a review of the literature some studies are presented, which focus on results of Balint work and changes in the participants of this group method. An empirical study of 436 physicians in Unterfranken shows that about 9% have participated in Balint work. Attitudes and motivations of the participants and setting and results of Balint work are studied as well as notions and knowledge of those, who have not done Balint work yet. The study demonstrates positive results in most of the participants.


Expression of TMPRSS2:ERG gene fusion in prostate cancer cells is an important prognostic factor for cancer progression.
The prostate-specific gene, TMPRSS2, is fused with the transcription factor gene, ERG in a high proportion of prostate cancers. However, the clinical significance of TMPRSS2:ERG gene fusion among prostate cancer patients is unknown. We assayed for the presence of the TMPRSS2:ERG gene fusion product among 26 patients who underwent surgery for clinically localized prostate cancer using RT-PCR and direct DNA sequencing, and evaluated its prognostic significance. All 26 patients had cancers of the same histologic grade (Gleason score 7). The fusion protein was present within prostate cancer tumor cells in eleven patients (42.3%). Nine patients experienced biochemical disease relapse (elevated PSA) after a mean follow-up of 12 months (range 1 to 48 months). Patients with the fusion protein had a significantly higher rate of recurrence (5-year recurrence rate 79.5%) compared to patients who lacked the fusion protein (five-year recurrence rate 37.5%, p = 0.009). The adjusted hazard ratio for disease relapse for patients with the fusion protein was 7.1 (95% C.I.: 1.1-45, p = 0.03) compared to patients without the fusion protein. In multivariate analysis, the presence of gene fusion was the single most important prognostic factor. Our study indicates that the expression of TMPRSS2:ERG fusion gene among prostate cancer patients treated with surgery is a strong prognostic factor for disease relapse, and may have important clinical implications.


Rules ventral prefrontal cortical axons use to reach their targets: implications for diffusion tensor imaging tractography and deep brain stimulation for psychiatric illness.
The ventral prefrontal cortex (vPFC) is involved in reinforcement-based learning and is associated with depression, obsessive-compulsive disorder, and addiction. Neuroimaging is increasingly used to develop models of vPFC connections, to examine white matter (WM) integrity, and to target surgical interventions, including deep brain stimulation. We used primate (Macaca nemestrina/Macaca fascicularis) tracing studies and 3D reconstructions of WM tracts to delineate the rules vPFC projections follow to reach their targets. vPFC efferent axons travel through the uncinate fasciculus, connecting different vPFC regions and linking different functional regions. The uncinate fasciculus also is a conduit for vPFC fibers to reach other cortical bundles. Fibers in the internal capsule are organized according to destination. Thalamic fibers from each vPFC region travel dorsal to their brainstem fibers. The results show regional differences in the trajectories of fibers from different vPFC areas. Overall, the medial/lateral vPFC position dictates the route that fibers take to enter major WM tracts, as well as the position within specific tracts: axons from medial vPFC regions travel ventral to those from more lateral areas. This arrangement, coupled with dorsal/ventral organization of thalamic/brainstem fibers through the internal capsule, results in a complex mingling of thalamic and brainstem axons from different vPFC areas. Together, these data provide the foundation for dividing vPFC WM bundles into functional components and for predicting what is likely to be carried at different points through each bundle. These results also help determine the specific connections that are likely to be captured at different neurosurgical targets.


Automatic extraction of upper-limb kinematic activity using deep learning-based markerless tracking during deep brain stimulation implantation for Parkinson's disease: A proof of concept study.
Optimal placement of deep brain stimulation (DBS) therapy for treating movement disorders routinely relies on intraoperative motor testing for target determination. However, in current practice, motor testing relies on subjective interpretation and correlation of motor and neural information. Recent advances in computer vision could improve assessment accuracy. We describe our application of deep learning-based computer vision to conduct markerless tracking for measuring motor behaviors of patients undergoing DBS surgery for the treatment of Parkinson's disease. Video recordings were acquired during intraoperative kinematic testing (N = 5 patients), as part of standard of care for accurate implantation of the DBS electrode. Kinematic data were extracted from videos post-hoc using the Python-based computer vision suite DeepLabCut. Both manual and automated (80.00% accuracy) approaches were used to extract kinematic episodes from threshold derived kinematic fluctuations. Active motor epochs were compressed by modeling upper limb deflections with a parabolic fit. A semi-supervised classification model, support vector machine (SVM), trained on the parameters defined by the parabolic fit reliably predicted movement type. Across all cases, tracking was well calibrated (i.e., reprojection pixel errors 0.016-0.041; accuracies >95%). SVM predicted classification demonstrated high accuracy (85.70%) including for two common upper limb movements, arm chain pulls (92.30%) and hand clenches (76.20%), with accuracy validated using a leave-one-out process for each patient. These results demonstrate successful capture and categorization of motor behaviors critical for assessing the optimal brain target for DBS surgery. Conventional motor testing procedures have proven informative and contributory to targeting but have largely remained subjective and inaccessible to non-Western and rural DBS centers with limited resources. This approach could automate the process and improve accuracy for neuro-motor mapping, to improve surgical targeting, optimize DBS therapy, provide accessible avenues for neuro-motor mapping and DBS implantation, and advance our understanding of the function of different brain areas.


[Prophylaxis and therapy of gastropathy caused by non-steroidal anti-inflammatory drugs].
The main disadvantage of the nonsteroidal anti-inflammatory drugs (NSAID) is their toxic effect on the gastroduodenal mucosa, which can cause life-threatening complications. We discuss the possibilities to prevent the harmful effect of the NSAIDs: the use of safer NSAIDs, and the co-therapy with protective mechanisms. Among the safer NSAIDs we discuss the preferentially selective, and the highly selective COX-2 inhibitors, and the new, nitric-oxide releasing NSAIDs. We consider the role of the mucosa-protecting mechanisms: antacids, sucralfate, H2 receptor antagonists, proton-pump inhibitors, and misoprostol. The most effective opportunities to protect the gastroduodenal mucosa--which is highly recommended for the high risk group of patients--are the usage of highly selective NSAIDs, or the concomitant therapy with proton-pump inhibitors or misoprostol. The value of the letter is diminished by it's side effects.


Multiple spontaneous coronary artery dissection associated with Trousseau's syndrome.
Spontaneous coronary artery dissection (SCAD) remains a rare cause of acute coronary syndrome. SCAD has been observed in three groups of patients: those with coronary atherosclerosis, women in the peripartum period, and an idiopathic group. SCAD may also be associated with some other conditions. Herein, we present a 57 year-old man who developed SCAD concomitant with Trousseau's syndrome secondary to colon adenocarcinoma.


Relative mutagenicities of gaseous nitrogen oxides in the supF gene of pSP189.
Gaseous nitric oxide (NO), an environmental pollutant found in cigarette smoke and diesel exhaust, has been shown to generate mutations in aerobic in vitro assays. The objective of this study was to identify which oxides of nitrogen, formed in the gaseous phase from NO, possess mutagenic activity. Samples of the plasmid pSP189, in 1 M HEPES buffer, pH 7.4, were exposed to preparations of nitrogen dioxide (NO2), dinitrogen trioxide (N2O3) or an air control. The gas mixtures were formed in a gas-tight syringe and were then introduced into 1 l flasks. The plasmid solution was introduced immediately afterwards. Transformation of Escherichia coli strain MBM7070 with the treated plasmids allowed analysis of mutation frequencies and the types of mutations induced in the target supF gene. The mutation frequency resulting from NO2 exposure was not different from that of the control. However, N2O3 produced a substantial number of mutations. The mutation frequency and the types of mutations were found to depend on the length of time that the gases were allowed to incubate in the syringe prior to introduction into the 1 l flasks (mutation frequency was maximal at approximately 2 min). The most prevalent mutations were AT-->GC transitions (68%), followed by GC-->AT transitions (30%), similar to previous findings when pure NO was bubbled through pSP189 solutions. These results suggest that it is N2O3, rather than NO2, that is the most likely source of mutagenic potential from gaseous nitrogen oxides.


Targeting Microglial Population Dynamics in Alzheimer's Disease: Are We Ready for a Potential Impact on Immune Function?
Alzheimer's disease (AD) is the most common form of dementia, affecting two-thirds of people with dementia in the world. To date, no disease-modifying treatments are available to stop or delay the progression of AD. This chronic neurodegenerative disease is dominated by a strong innate immune response, whereby microglia plays a central role as the main resident macrophage of the brain. Recent genome-wide association studies (GWASs) have identified single-nucleotide polymorphisms (SNPs) located in microglial genes and associated with a delayed onset of AD, highlighting the important role of these cells on the onset and/or progression of the disease. These findings have increased the interest in targeting microglia-associated neuroinflammation as a potentially disease-modifying therapeutic approach for AD. In this review we provide an overview on the contribution of microglia to the pathophysiology of AD, focusing on the main regulatory pathways controlling microglial population dynamics during the neuroinflammatory response, such as the colony-stimulating factor 1 receptor (CSF1R), its ligands (the colony-stimulating factor 1 and interleukin 34) and the transcription factor PU.1. We also discuss the current therapeutic strategies targeting proliferation to modulate microglia-associated neuroinflammation and their potential impact on peripheral immune cell populations in the short and long-term. Understanding the effects of immunomodulatory approaches on microglia and other immune cell types might be critical for developing specific, effective, and safe therapies for neurodegenerative diseases.


Effect of genetic strain and gender on age-related changes in body composition of the laboratory rat.
Body fat serves as a storage compartment for lipophilic pollutants and affects the pharmacokinetics of many toxic chemicals. Understanding how body fat varies with gender, strain, and age may be essential for development of experimental models to study mechanisms of toxicity. Nuclear magnetic resonance (NMR)-based analysis serves as a noninvasive means of assessing proportions of fat, lean, and fluid in rodents over their lifetime. The aim of this study was to track changes in body composition of male and female Long-Evans (LE), Sprague-Dawley (SD), Fischer (F334), and Brown Norway (BN) rats from postweaning over a >2-yr period. Percent fat of preweaned LE and SD rats was markedly higher compared to the other strains. LE and SD strains displayed marked increases in body fat from weaning to 8 mo of age. Postweaned F344 male and females showed relatively low levels of percent fat; however, at 2 yr of age percent fat of females was equal to that of SD and LE in females. BN rats showed the highest levels of lean tissue and lowest levels of fat. Percent fat of the BN strain rose at the slowest rate as they aged. Percent fluid was consistently higher in males for all strains. Females tended to have higher percent fat than males in LE, SD, and F344 strains. Assessing changes in body fat as well as lean and fluid of various strains of male and female rats over their lifetime may prove useful in many research endeavors, including pharmacokinetics of lipophilic toxicants, mechanisms underlying obesity, and metabolic disorders.


Printable Smart Materials and Devices: Strategies and Applications.
Smart materials are a kind of functional materials which can sense and response to environmental conditions or stimuli from optical, electrical, magnetic mechanical, thermal, and chemical signals, etc. Patterning of smart materials is the key to achieving large-scale arrays of functional devices. Over the last decades, printing methods including inkjet printing, template-assisted printing, and 3D printing are extensively investigated and utilized in fabricating intelligent micro/nano devices, as printing strategies allow for constructing multidimensional and multimaterial architectures. Great strides in printable smart materials are opening new possibilities for functional devices to better serve human beings, such as wearable sensors, integrated optoelectronics, artificial neurons, and so on. However, there are still many challenges and drawbacks that need to be overcome in order to achieve the controllable modulation between smart materials and device performance. In this review, we give an overview on printable smart materials, printing strategies, and applications of printed functional devices. In addition, the advantages in actual practices of printing smart materials-based devices are discussed, and the current limitations and future opportunities are proposed. This review aims to summarize the recent progress and provide reference for novel smart materials and printing strategies as well as applications of intelligent devices.


Gram-scale solvothermal synthesis of Fe-doped CuCoO2 nanosheets and improvement of the oxygen evolution reaction performance.
In this work, we used Cu-BTC-IPA and Co(NO3)2·6H2O as precursors to synthesize CuCoO2 (CCO) nanocrystals with a suitable crystal phase, morphology and high yield by changing the process parameters, such as reactant concentration, reactant ratio, mineralizer dosage, and the type of polyvinylpyrrolidone surfactant. In addition, the effects of different concentrations (1 at%, 3 at%, 5 at%) of Fe doping on the crystal structure and oxygen evolution reaction (OER) performance of CCO were studied. The experimental results show that Fe ions are uniformly doped into the lattice to replace the A-site (Cu+) position, which not only reduces the grain size of CCO, but also increases its specific surface area. We further employed the density functional theory (DFT) method to simulate the OER process of transition metal Fe-doped CCO (A-site substitution) and proposed that Fe doping can reduce the Gibbs free energy of each step and promote the formation of each intermediate, thereby improving its OER catalytic performance. In 1.0 M KOH electrolyte, the 3 at% Fe-doped CCO (Ni@3FCCO) electrode has the best OER performance (η10 = 369 mV, Tafel slope = 69 mV dec-1), and the required overpotential to attain 10 mA cm-2 slightly increased (∼30.2 mV) after 18 hours of continuous OER. The crystal morphology and chemical composition did not change significantly before and after the long-term OER test, indicating that the 3FCCO nanosheets have good OER activity and stability. We have proposed two reasons for the significant improvement of OER performance for Fe-doped CCO nanosheets: (1) the partial substitution of Cu cations by Fe cations not only regulates the electronic structure of CCO, making the catalytically active center no longer a single Co site, but also contains the Fe site, thus increasing the number of overall active sites; (2) the synergistic effect between Fe cations and Co cations in the OER process could enhance the activity of a single active site.


MORPHOLOGICAL EVOLUTION OF THE DROSOPHILA VIRILIS SPECIES GROUP AS ASSESSED BY RATE TESTS FOR NATURAL SELECTION ON QUANTITATIVE CHARACTERS.
Two rate tests for assessing natural selection on quantitative traits are discussed for their usefulness in macroevolutionary and adaptational studies. The underlying assumptions and parameter estimation for the constant-heritability (CH) and mutation-drift-equilibrium (MDE) models, which are the bases for these tests, are discussed. The purpose of these rate tests is to determine whether morphological change has occurred either too fast to be explained by neutral drift, which suggests directional selection, or too slow, which suggests stabilizing selection. Previous formulations of these rate tests have ignored the phylogenetic component. Several models of evolution are considered to help account for phylogeny in the context of rate tests. The MDE rate test for stabilizing selection was performed on nine morphological characters among several species of the Drosophila virilis species group. These tests can be interpreted to suggest that stabilizing selection has probably been a major factor in producing the observed similarity among the Drosophila species examined.


Metabolic transit of N(ε)-carboxymethyl-lysine after consumption of AGEs from bread crust.
Our aim was to investigate carboxymethyl-lysine (CML) intake and excretion after feeding rats with diets containing advanced glycation end-products (AGEs) from bread crust (BC) or its soluble or insoluble fractions, and to identify the factors responsible for the effects observed. CML in serum and different tissues was measured to detect possible accumulations. For 88 days, weanling rats were fed with either a control diet or one containing BC, or its soluble low molecular weight (LMW), soluble high molecular weight (HMW) or insoluble fractions. In the last week of the assay, faeces and urine were collected daily and stored as a 1 week pool. After sacrifice, blood was drawn to obtain serum and some organs were removed. CML analysis was performed by HPLC/MS/MS in diets, faeces, urines, serum and tissues. Faecal excretion of CML was strongly influenced by dietary CML levels and represents the major route of excretion (i.e. 33.2%). However, the urinary elimination of CML was probably limited or saturated, especially when more complex compounds were present in the diet. BC consumption increased CML in the cardiac tissue (170 ± 18 vs. 97 ± 3 μmol per mol lysine for BC and control groups), which correlated with the CML intake. The levels of this AGE in bone were unaffected by the dietary treatment, but in tail tendons CML was greatly increased in the animals that consumed the BC diet (102 ± 13 vs. 51 ± 8 μmol per mol lysine for BC and control groups, P = 0.006), which was associated with the intake of soluble LMW compounds present in BC. Despite the CML accumulation detected in different tissues, serum levels of protein-bound CML were unchanged, indicating the importance of measuring the free CML in this fluid as a real index of dietary CML.


Fine and ultrafine particles of the diet: influence on the mucosal immune response and association with Crohn's disease.
Crohn's disease is a modern Western disease characterised by transmural inflammation of the gastrointestinal tract. It is of unknown aetiology, but evidence suggests that it results from a combination of genetic predisposition and environmental factors. Bacterial-sized microparticles (0.1-1.0 microm) are potent adjuvants in model antigen-mediated immune responses and are increasingly associated with disease. Microparticles of TiO2 and aluminosilicate accumulate in macrophages of human gut-associated lymphoid tissue where the earliest signs of lesions in Crohn's disease are observed. Dietary microparticles are of endogenous or exogenous origin. Endogenous microparticles dominate and are calcium phosphate (most probably hydroxyapatite), which precipitates in the lumen of the mid-distal gastrointestinal tract due to secretion of Ca and phosphate in the succus entericus. Exogenous dietary microparticles are contaminants (soil and/or dust) and food additives. TiO2, for example, is a food colourant, and aluminosilicates are anti-caking agents, although some aluminosilicates occur as natural contaminants. Food additives alone account for ingestion of approximately 10(12) particles/person per d. Possible mechanisms for the role of exogenous and endogenous dietary microparticles in promoting toleragenic or immune responses of gastrointestinal mucosal phagocytosis are discussed. In a double-blind randomised pilot study we have shown that a diet low in Ca and exogenous microparticles appears to alleviate the symptoms of ileal Crohn's disease, with a significant (P= 0.002) improvement in the Crohn's disease activity index. A multi-centre trial and further mechanistic studies at the cellular level are underway.


Microarray analysis of amphotericin B-treated THP-1 monocytic cells identifies unique gene expression profiles among lipid and non-lipid drug formulations.
The effects of deoxycholate amphotericin B (DAmB), amphotericin B lipid complex (ABLC), and amphotericin B colloidal dispersion (ABCD) on mRNA profiles from 218 genes in treated THP-1 monocytes were compared. Sixty-one genes were up-regulated and 8 were down-regulated by one or more of the AmB formulations. Fifty-three genes were up-regulated by DAmB while 24 and 18 genes were up-regulated by ABCD and ABLC, respectively. DAmB and ABCD up-regulated many pro-inflammatory genes, whereas ABLC did not. All three formulations up-regulated multiple categories of genes including the anti-apoptosis gene BIRC3 and the chemokine CXCL9 (MIG). Based on representative genes, DAmB activated more signaling pathways than ABCD or ABLC. Quantitative real time PCR confirmed array up-regulation of representative pro-inflammatory genes IL-8, CCL20 (MIP-3alpha), and CXCL2 (MIP-2alpha). Our study represents the first larger scale comparative gene expression profiling which may provide additional rationales for clinical side effects of each amphotericin B formulation.


Interstitial fibrosis in papillary thyroid microcarcinoma and its association with biological behavior.
Previous studies have revealed that fibrosis may affect the biological behavior of tumors, however associated research on papillary thyroid microcarcinoma (PTMC) is rare. The aim of the present study was to explore the association between interstitial fibrosis (IF) and the biological behavior of PTMC. In the present study, a total of 511 consecutive cases of PTMC were evaluated for the presence of IF and its association with clinical parameters and pathologic biomarkers. IF was identified in 340 of the 511 consecutive cases and it was significantly associated with the age (P=0.033), tumor diameter (P=0.017) and lymphocytic metastasis (P<0.001) of the patient. There were significantly more female in the fibrosis group than in fibrosis-absent group (P=0.024). In the analysis of clinical biomarkers using immunohistochemistical staining, IF was significantly associated with cytokeratin 19 (P=0.008) and galectin-3 (P=0.022). Analysis of patient outcomes indicated that IF was an independent prognostic factor of recurrence (hazard ratio = 2.181; 95% confidence interval = 1.163-4.090; P=0.015). These findings suggest that the combined effect of a patient's age, sex and tumor size may potentially contribute to fibrotic lesions and IF was a factor contributing to poor prognosis in patients with PTMC.


Inhibitory action of silibinin on low density lipoprotein oxidation.
Low density lipoprotein (LDL) oxidation and smooth muscle cell growth represent key events in atherogenesis. Any mean to reduce these two phenomena may decrease the risk of coronary artery disease and atherosclerosis in general. The effects of silibinin (CAS 22888-70-6) on LDL oxidation and proliferation of vascular smooth muscle cells were evaluated in vitro. Silibinin (50-200 mumol/l) prolonged the lag times of both LDL autooxidation and oxidation by copper by > 50%, as assessed by recordings of diene formation. However, silibinin (up to 500 mumol/l) did not interfere with LDL-stimulated radiolabeled thymidine incorporation. These findings indicate that silibinin, apart from its hepatoprotective effects, has inhibitory properties on LDL oxidation in vitro. Therefore silibinin might represent a novel tool in the prevention and therapy of atherosclerosis.


Fatty acid kinetics and carbohydrate metabolism during electrical exercise in spinal cord-injured humans.
Motor center activity and reflexes from contracting muscle have been shown to be important for mobilization of free fatty acids (FFA) during exercise. We studied FFA metabolism in the absence of these mechanisms: during involuntary, electrically induced leg cycling in individuals with complete spinal cord injury (SCI). Healthy subjects performing voluntary cycling served as controls (C). Ten SCI (level of injury: C5-T7) and six C exercised for 30 min at comparable oxygen uptake rates (approximately 1 l/min), and [1-14C]palmitate was infused continuously to estimate FFA turnover. From femoral arteriovenous differences, blood flow, muscle biopsies, and indirect calorimetry, leg substrate balances as well as concentrations of intramuscular substrates were determined. Leg oxygen uptake was similar in the two groups during exercise. In SCI, but not in C, plasma FFA and FFA appearance rate fell during exercise, and plasma glycerol increased less than in C (P < 0.05). Fractional uptake of FFA across the working legs decreased from rest to exercise in all individuals (P < 0.05) but was always lower in SCI than in C (P < 0.05). From rest to exercise, leg FFA uptake increased less in SCI than in C subjects (14 +/- 3 to 57 +/- 20 vs. 41 +/- 13 to 170 +/- 57 micromol x min(-1) x leg(-1); P < 0.05). Muscle glycogen breakdown, leg glucose uptake, carbohydrate oxidation, and lactate release were higher (P < 0.05) in SCI than in C during exercise. Counterregulatory hormonal changes were more pronounced in SCI vs. C, whereas insulin decreased only in C. In conclusion, FFA mobilization, delivery, and fractional uptake are lower and muscle glycogen breakdown and glucose uptake are higher in SCI patients during electrically induced leg exercise compared with healthy subjects performing voluntary exercise. Apparently, blood-borne mechanisms are not sufficient to elicit a normal increase in fatty acid mobilization during exercise. Furthermore, in exercising muscle, FFA delivery enhances FFA uptake and inhibits carbohydrate metabolism, while carbohydrate metabolism inhibits FFA uptake.


Biological mechanisms of action of novel C-10 non-acetal trioxane dimers in prostate cancer cell lines.
The mechanisms of action of three C-10 non-acetal trioxane dimers (TDs) were examined in human (LNCaP) and mouse (TRAMP-C1A and -C2H) prostate cancer cell lines. 1 (AJM3/23), 2 (GHP-TM-III-07w), and 3 (GHP-KB-06) inhibited cell growth with 3 being the most potent in C1A (GI50 = 18.0 nM), C2H (GI50 = 17.0 nM), and LNCaP (GI50 = 17.9 nM) cells. In comparison to a standard cytotoxic agent such as doxorubicin (GI50 = 45.3 nM), 3 (GI50 = 17.9 nM) inhibited LNCaP cell growth more potently. TDs induced G0/G1 cell cycle arrest in LNCaP cells and decreased cells in the S phase. These changes correlated with modulation of G1 phase cell cycle proteins including decreased cyclin D1, cyclin E, and cdk2 and increased p21waf1 and p27Kip1. TDs also promoted apoptosis in LNCaP cells with increased expression of proapoptotic bax. These results demonstrate that TDs are potentially useful agents that warrant further preclinical development for treatment of prostate cancer.


The effect of diabetic control on the width of skeletal-muscle capillary basement membrane in patients with Type I diabetes mellitus.
We studied the relation between the control of blood glucose and the width of skeletal-muscle capillary basement membrane in 23 insulin-dependent (Type I) diabetic patients. After initial measurement of levels of glycosylated hemoglobin and width of skeletal-muscle capillary basement membrane, the patients were divided into two groups: an experimental group of 13 patients who were treated with continuous subcutaneous insulin infusion, and a control group of 10 patients who continued to receive conventional treatment--usually two injections of insulin daily. After two years, the experimental group had a significant decrease in glycosylated hemoglobin levels as compared with base-line values (mean +/- S.E.M., 7.6 +/- 0.4 vs 10.2 +/- 0.7 per cent; P less than 0.001), reflecting improved control of blood glucose, and a significant reduction in the width of skeletal-muscle capillary basement membrane (1293 +/- 68 vs. 1717 +/- 182 A; P less than 0.05). The control group of patients had no significant change in their levels of glycosylated hemoglobin or in the width of their skeletal-muscle capillary basement membranes. If changes in the capillaries in skeletal muscle parallel those in the capillaries in retinal or renal tissue, then meticulous control of blood glucose may be beneficial over time in preventing the microvascular complications of diabetes.


The histologic spectrum of Barrett's esophagus.
To define the histology of the columnarlined esophagus, we obtained esophageal biopsies from various levels with manometric control from 11 patients. There were three types of columnar epithelia above the lower esophageal sphincter: atrophic gastric-fundic-type epithelium with parietal and chief cells; junctional-type epithelium with cardiac mucous glands; and distinctive specialized columnar epithelium with a villiform surface, mucous glands and intestinal-type goblet cells. When present, specialized columnar epithelium was always the most proximal, and gastric fundic epithelium the most distal epithelium. Junctional epithelium was interposed between gastric fundic and specialized columnar or squamous epithelium. Four patients had unequivocal esophagitis in squamous epithelium, but its presence and severity did not correlate with inflammation in or length or type of distal columnar epithelium. Histoligic study of the columnar-lined esophagus demonstrated a spectrum of epithelial patterns. This heterogeneity helps to explain prior discrepant reports.


A randomised trial comparing 5-FU with 5-FU plus cisplatin in advanced pancreatic carcinoma.
Chemotherapy is moderately efficient as a treatment for pancreatic adenocarcinoma, but patient survival and quality of life has improved with this modality in some trials. In a previous phase II trial, 5-fluorouracil (5-FU) plus cisplatin (FUP) yielded a 26.5% response rate and a 29% survival rate at 1 year. The present study aimed to compare FUP with 5-FU alone, which was the control arm in former Mayo Clinic trials. Patients with untreated cytologically or histologically proven metastatic or locally advanced adenocarcinoma of the pancreas were deemed measurable or evaluable. Chemotherapy regimens consisted of a control FU arm (5-FU 500 mg/m(2)/day for 5 days) and the investigational FUP arm (continuous 5-FU 1000 mg/m(2)/day for 5 days plus cisplatin 100 mg/m(2) on day 1 or day 2). In both arms, chemotherapy was repeated at day 29. Two-hundred and seven patients from 18 centres were randomised: 103 in the FU arm and 104 in FUP arm. Treatment arms were balanced with respect to performance status grade 0-1 (83% versus 86%, respectively) and the presence of metastases (92% versus 89%, respectively). The median number of cycles administered was two in both arms (range 0-14). Five patients did not receive any chemotherapy and 45 received only one cycle. Toxicity (WHO grade 3-4) was lower with FU than with FUP (20% versus 48%, P <0.001), as was neutropenia (6% versus 23%), vomiting (4% versus 17%) and toxicity-related deaths (one versus four early in the trial). The response rate was low in both arms, but superior in the FUP arm: 12% versus 0% (intention-to-treat analysis, P <0.01). The survival rates at 6 months were 28% and 38% for the FU and FUP arms, respectively, and 1-year survival rates were 9% and 17% (log-rank test, P = 0.10). One-year progression-free survival was 0% with FU versus 10% with FUP (log-rank test, P = 0.0001). In advanced pancreatic carcinomas with a poor prognosis, FUP was superior to FU in terms of response and progression-free survival, but not in terms of overall survival. The low response rate is partly related to the number of patients who received only one cycle of chemotherapy. A more effective, better tolerated version of this FUP combination is needed.


Stochastic chemical kinetics and the total quasi-steady-state assumption: application to the stochastic simulation algorithm and chemical master equation.
Recently the application of the quasi-steady-state approximation (QSSA) to the stochastic simulation algorithm (SSA) was suggested for the purpose of speeding up stochastic simulations of chemical systems that involve both relatively fast and slow chemical reactions [Rao and Arkin, J. Chem. Phys. 118, 4999 (2003)] and further work has led to the nested and slow-scale SSA. Improved numerical efficiency is obtained by respecting the vastly different time scales characterizing the system and then by advancing only the slow reactions exactly, based on a suitable approximation to the fast reactions. We considerably extend these works by applying the QSSA to numerical methods for the direct solution of the chemical master equation (CME) and, in particular, to the finite state projection algorithm [Munsky and Khammash, J. Chem. Phys. 124, 044104 (2006)], in conjunction with Krylov methods. In addition, we point out some important connections to the literature on the (deterministic) total QSSA (tQSSA) and place the stochastic analogue of the QSSA within the more general framework of aggregation of Markov processes. We demonstrate the new methods on four examples: Michaelis-Menten enzyme kinetics, double phosphorylation, the Goldbeter-Koshland switch, and the mitogen activated protein kinase cascade. Overall, we report dramatic improvements by applying the tQSSA to the CME solver.


Sialidosis and galactosialidosis: chromosomal assignment of two genes associated with neuraminidase-deficiency disorders.
The inherited human disorders sialidosis and galactosialidosis are the result of deficiencies of glycoprotein-specific alpha-neuraminidase (acylneuraminyl hydrolase, EC 18.104.22.168; sialidase) activity. Two genes were determined to be necessary for expression of neuraminidase by using human-mouse somatic cell hybrids segregating human chromosomes. A panel of mouse RAG-human hybrid cells demonstrated a single-gene requirement for human neuraminidase and allowed assignment of this gene to the (pter----q23) region of chromosome 10. A second panel of mouse thymidine kinase (TK)-deficient LM/TK- -human hybrid cells demonstrated that human neuraminidase activity required both chromosomes 10 and 20 to be present. Analysis of human neuraminidase expression in interspecific hybrid cells or polykaryocytes formed from fusion of mouse RAG (hypoxanthine/guanine phosphoribosyltransferase deficient) or LM/TK- cell lines with human sialidosis or galactosialidosis fibroblasts indicated that the RAG cell line complemented the galactosialidosis defect, but the LM/TK- cell line did not. This eliminates the requirement for this gene in RAG-human hybrid cells and explains the different chromosome requirements of these two hybrid panels. Fusion of LM/TK- cell hybrids lacking chromosome 10 or 20 (phenotype 10+,20- and 10-,20+) and neuraminidase-deficient fibroblasts confirmed by complementation analysis that the sialidosis disorder results from a mutation on chromosome 10, presumably encoding the neuraminidase structural gene. Galactosialidosis is caused by a mutation in a second gene required for neuraminidase expression located on chromosome 20.


Routine obstetric ultrasound: effectiveness of cross-sectional screening for fetal growth retardation.
Sonographic detection of fetal growth retardation was evaluated on a sample of 778 patients routinely scanned over a 16-month period. From the 29th week to term, the sensitivity of abdominal circumference measurement progressively increased from 41% to 88% (P less than 0.01). Higher rates were achieved by using the biparietal diameter but false-positive diagnoses were twice as frequent as with the abdominal measurements. When the limitations in precision of abdominal circumference were considered, the screening rate of this parameter increased about 16%. Moreover, 50% of the false-negative diagnoses were made on fetuses weighing between the fifth and 10th centiles at birth. These diagnostic indices for biparietal diameter and abdominal circumference evaluated on a retrospective series proved to be lower than those reported for the prospective selected series. However, the results yielded by routine ultrasound were better than those obtained with clinical or traditional biochemical methods. These findings underline the importance of sonography not only in serial control of high-risk pregnancies but also in cross-sectional screening programs.


Determination of bioavailability and identification of collagen peptide in blood after oral ingestion of gelatin.
Gelatin has long been widely used in foods, pharmaceuticals, cosmetics, and other products. However, there are few reports on its bioavailability and bioavailable forms. In this study, the bioavailability of gelatin was indirectly evaluated by the determining the bioavailability of total hydroxyproline in gelatin using a pharmacokinetic method after oral administration to rats. The relative and absolute bioavailability of gelatin were 74.12% and 85.97%, respectively. The amino acid profile of plasma indicated that 41.91% of the digested gelatin was absorbed from the intestine in the form of peptide, and there was a good linear correlation between the absorbed amount of an amino acid and its content in gelatin (R(2) = 0.9566). Moreover, 17 types of collagen peptide were purified by multi-step chromatography and identified with ultra-performance liquid chromatography-electrospray ionisation-mass spectrometry. Gelatin had high oral bioavailability. Nearly half of digested gelatin was absorbed from the intestine in the form of various collagen peptides.


Peroxisome proliferator-activated receptors and the control of inflammation.
Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors which form a subfamily of the nuclear receptor gene family. This subfamily consists of three isotypes, alpha (NR1C1), gamma (NR1C3), and beta/delta (NRC1C2) with a differential tissue distribution. PPARalpha is expressed primarily in tissues with a high level of fatty acid catabolism such as liver, brown fat, kidney, heart and skeletal muscle. PPARbeta is ubiquitously expressed, and PPARgamma has a restricted pattern of expression, mainly in white and brown adipose tissues, whereas other tissues such as skeletal muscle and heart contain limited amounts. Furthermore, PPARalpha and gamma isotypes are expressed in vascular cells including endothelial and smooth muscle cells and macrophages/foam cells. PPARs are activated by ligands, such as naturally occurring fatty acids, which are activators of all three PPAR isotypes. In addition to fatty acids, several synthetic compounds, such as fibrates and thiazolidinediones, bind and activate PPARalpha and PPARgamma, respectively. In order to be transcriptionally active, PPARs need to heterodimerize with the retinoid-X-receptor (RXR). Upon activation, PPAR-RXR heterodimers bind to DNA specific sequences called peroxisome proliferator-response elements (PPRE) and stimulate transcription of target genes. PPARs play a critical role in lipid and glucose homeostasis, but lately they have been implicated as regulators of inflammatory responses. The first evidence of the involvement of PPARs in the control of inflammation came from the PPARalpha null mice, which showed a prolonged inflammatory response. PPARalpha activation results in the repression of NF-kappaB signaling and inflammatory cytokine production in different cell-types. A role for PPARgamma in inflammation has also been reported in monocyte/macrophages, where ligands of this receptor inhibited the activation of macrophages and the production of inflammatory cytokines (TNFalpha, interleukin 6 and 1beta), although part of the anti-inflammatory effects of these ligands seems to be mediated by a mechanism not involving PPARgamma. All these findings suggest a role of PPARs in the control of the inflammatory response with potential therapeutic applications in inflammation-related diseases.


Hb Amsterdam-A1 [α32(B13)Met→Ile; HBA1: c.99G>A]: A Hyperunstable Variant Due to a New Mutation on the α1 Gene.
Patients with hyperunstable α chain variants usually present with a thalassemic, rather than hemolytic, phenotype. Electrophoretic, ion exchange and reverse phase separations usually fail to detect the variant and when DNA sequencing identifies a 'silent' substitution it is usually presumed to be hyperunstable. We report the identification of such a variant, α32(B13)Met→Ile; HBA1: c.99G>A, arising from a new mutation on the α1 gene. The hemoglobin (Hb) was unequivocally detected by the isopropanol stability test and confirmed as hyperunstable by mass spectrometry (MS) of the precipitate and lysate, which showed proportions of 55% and 2.5% of α chains, respectively. The instability appears to be driven by perturbation of globin-heme, and possibly α1β1 subunit, interactions.


[Comparison of ELIFA and IGM-immunocapture technics in the diagnosis of 50 cases of congenital toxoplasmosis].
The authors have compared the diagnostic value in congenital toxoplasmosis of two recently developed immunological techniques, Enzyme Linked immunofiltration Assay (ELIFA) and IgM-immunocapture (or immunosorbent agglutination assay). The study involved 50 children suffering from congenital toxoplasmosis and 300 unscathed control children, whose mothers suffered from toxoplasmosis during their pregnancies. The ELIFA technique showed a sensitivity of 74 p. cent at the end of the first month after birth (day 30) and 84 p. cent after two months (day 60), whereas the IgM-immunocapture technique gave a sensitivity of 64 p. cent at day 30 and 70 p. cent at day 60. Both methods were at least 95 p. cent specific. The two techniques seem to be complementary to each other, and their association allowed the diagnosis to be confirmed in 80 p. cent of cases at day 30 and in 90 p. cent of cases at day 60. A serum sample taken on the tenth day after birth (day 10) was important both to eliminate the rare cases (5 p. cent) of transplacental transmission of maternal IgM antibodies and to detect transient neonatal synthesis of IgG or IgM toxoplasmic antibodies.


Permeability of the ovarian follicle of Aedes aegypti mosquitoes.
The passage of tracers of various molecular weights into resting and vitellogenic ovarian follicles of Aedes aegypti mosquitoes was studied ultrastructurally. The outermost layer of the follicular sheath (the basement lamina) is a coarse mechanical filter. It is freely permeable to particles with molecular weights ranging from 12,000 to 500,000 (i.e. cytochrome c, peroxidase, hemoglobin, catalase, ferritin, immunoglobulin (IgG)-peroxidase, iron dextran and Thorotrast) that have dimensions less than 110 A. Molecules as large as carbon (300-500 A) are totally excluded. Whereas proteins and polysaccharide tracers permeate the basement lamina with apparent ease, certain inert particles (e.g. Thorotrast, Fellows-Testager Div., Fellows Mfg. Co., Inc., Detroit, Mich.) penetrate more slowly. With respect to the tracers tested, resting follicles are as permeable as vitellogenic follicles. The follicle epithelium of resting or vitellogenic follicles is penetrated by narrow intercellular channels. Our observations suggest that these spaces are lined with mucopolysaccharide material. After permeating the basement lamina, exogenous tracers fill these channels, while the bulk of material accumulates in the perioocytic space. Within 3 hr after imbibing blood, the pinocytotic mechanism of the oocyte is greatly augmented. Pinocytosis is not selective with regard to material in the perioocytic space, since double tracer studies show that exogenous compounds are not separated, but are incorporated into the same pinocytotic vesicle. During later stages of vitellogenesis, 36-48 hr after the blood-meal, the pinocytotic mechanism of the oocyte is diminished. Simultaneously, the intercellular channels become occluded by desmosomes, and the vitelline membrane plaques separate the oocyte and follicle epithelium.


Microvessel density in patients with gastrointestinal stromal tumors: A systematic review and meta-analysis.
Gastrointestinal stromal tumors (GISTs) are considered the most common mesenchymal tumors of the gastrointestinal tract. Microvessel density (MVD) constitutes a direct method of vascularity quantification and has been associated with survival rates in multiple malignancies. To appraise the effect of MVD on the survival of patients with GIST. This study adhered to Systematic reviews and Meta-Analyses guidelines and the Cochrane Handbook for Systematic Reviews of Interventions. Electronic scholar databases and grey literature repositories were systematically screened. The Fixed Effects or Random Effects models were used according to the Cochran Q test. In total, 6 eligible studies were identified. The pooled hazard ratio (HR) for disease free survival (DFS) was 8.52 (95%CI: 1.69-42.84, P = 0.009). The odds ratios of disease-free survival between high and low MVD groups at 12 and 60 mo did not reach statistical significance. Significant superiority of the low MVD group in terms of DFS was documented at 36 and 120 mo (OR: 8.46, P < 0.0001 and OR: 22.71, P = 0.0003, respectively) as well as at metastases rate (OR: 0.11, P = 0.0003). MVD significantly correlates with the HR of DFS and overall survival rates at 36 and 120 mo. Further prospective studies of higher methodological quality are required.


Successful surgical employment of Impella recovery system for right ventricular failure after previous aortic valve replacement.
A 58-year-old woman underwent aortic valve replacement. On the second postoperative day the patient referred a sharply chest pain, and an emergent coronary angiography revealed total occlusion of the right coronary artery. An intra-aortic ballon pump was placed and the patient underwent emergent off-pump coronary revascularization of the right coronary artery. Five hours later, due to unstable hemodynamic the extracorporeal membrane oxygenation was implanted without improvement of the right ventricular (RV) function. Then we decided to implant the Impella Right Direct (RD). After 9 days of Impella's insertion the RV was recovered and the device was successfully explanted. After 16 days of Impella explanted the patient was discharged. This case suggest that implantation of Impella RD is clinically feasible, associated with hemodynamic improvement, and facilitate successful bridge-to-recovery in patients with post-cardiotomy RV failure due to myocardial infarction unresponsive to coronary artery bypass grafting, maximal medical therapy, contrapulsation and extracorporeal membrane oxygenation.


Temporal trends of human immunodeficiency virus type 1 (HIV-1) infection among inmates entering a statewide prison system, 1985-1987.
Acquired immune deficiency syndrome (AIDS) became the leading cause of death among Maryland State prisoners in 1985. To identify the prevalence, risk factors, and temporal trends for infection with the human immunodeficiency virus type 1 (HIV-1) in the statewide prison system, excess sera were obtained from incoming male inmates during specified periods between April and June 1985, 1986, and 1987. Correctional medical personnel also provided demographic variables of age, race, offense category, sentence, jurisidiction, and an indicator of intravenous drug use. Once rendered anonymous, specimens were assayed for antibody to HIV-1, using ELISA and Western blot techniques. For data from April to June 1985, 1986, and 1987, the crude prevalence of anti-HIV-1 was 7.1, 7.7, and 7.0%, respectively. Although one-third of incoming inmates were identified as intravenous drug users (IVDUs), the drug use variable was missing for 70% of the 1985 sample, and 40% of the 1986 sample. Several strategies were used to examined temporal trends in the context of missing data. Univariate analyses suggested no substantial change over time for either HIV-1 seroprevalence or risk of infection among IVDUs.(ABSTRACT TRUNCATED AT 250 WORDS)


[Palliative surgical treatment of spastic paralysis in the upper extremity].
In this paper we review the main studies conducted on therapy applied to the bony and soft parts in spastic paralysis of the upper extremity. Spasticity presents muscular hypertonia and hyperexcitability of the stretch reflex, which are typical of upper motoneuron syndrome. Physiopathologically, spasticity is due to the medullar and supramedullar alteration of the afferent and efferent pathways. Treatment is multidisciplinary and involves the collaboration of rehabilitators, neurophysiologists, neurologists, paediatricians, orthopaedic surgeons and psychologists, who all contribute with their different therapeutic aspects and characteristics (which can be pharmacological, peripheral neurological blockages, surgical, etc.). The characteristic posture of the upper extremities in spastic cerebral palsy is the inward rotation of the shoulder, flexion of the elbow and pronated forearm, and the deformity of the fingers (swan-neck and thumbs-in-palm). The primary objectives in these patients will be to improve communication with their surroundings, perform activities of daily living, increase mobility and walking. The surgical treatment applied by orthopaedic surgeons in the upper extremities are aimed at achieving an enhanced adaptive functionality rather than morphological normality. Factors to be taken into account include age, voluntary control over muscles and joints, level of severity of the spasticity (Ashworth scale) and stereognostic sensitivity. In general, on soft parts we will use procedures such as dehiscence or lengthening of the flexor muscles of the shoulder and elbow or of the adductor of the thumb; transfer of the pronators in order to adopt the supinating function or of the flexors so as to reinforce the extensors of the forearm, and capsulodesis or tenodesis in the hand. The bony procedures will consist in derotational osteotomies of the humerus and radius and arthrodesis in the wrist or in the metacarpophalangeal joints of the thumb, depending on whether there is greater rigidity or age in the former cases or instability in the latter.


Development of a Virtual Cell Model to Predict Cell Response to Substrate Topography.
Cells can sense and respond to changes in the topographical, chemical, and mechanical information in their environment. Engineered substrates are increasingly being developed that exploit these physical attributes to direct cell responses (most notably mesenchymal stem cells) and therefore control cell behavior toward desired applications. However, there are very few methods available for robust and accurate modeling that can predict cell behavior prior to experimental evaluations, and this typically means that many cell test iterations are needed to identify best material features. Here, we developed a unifying computational framework to create a multicomponent cell model, called the "virtual cell model" that has the capability to predict changes in whole cell and cell nucleus characteristics (in terms of shape, direction, and even chromatin conformation) on a range of cell substrates. Modeling data were correlated with cell culture experimental outcomes in order to confirm the applicability of the virtual cell model and demonstrating the ability to reflect the qualitative behavior of mesenchymal stem cells. This may provide a reliable, efficient, and fast high-throughput approach for the development of optimized substrates for a broad range of cellular applications including stem cell differentiation.


Adhesive strength between fiber-reinforced composites and veneering composites and fracture load of combinations of these materials.
This study examined the influence of the adhesive strength between fiber-reinforced composites (FRC) and veneering composites on the fracture load of combinations of these materials. Six materials were used. An experimental material called BR-100, Vectris, and FibreKor were the types of FRC. Estenia, Targis, and Sculpture were used as veneering composites. With the Estenia/BR-100 combination, the surface of the FRC was subjected to three different conditions before veneering. Ten specimens of each combination were fabricated and divided into two groups: One group was stored in 37 degrees C distilled water for 24 hours, and the other was thermocycled (4 degrees C/60 degrees C, 10,000 cycles). Adhesive strength between FRCs and veneering composites was determined using the compressive shear strength test. In addition, fracture loads of laminate specimens were determined. Good adhesive strength was obtained by leaving an unpolymerized layer on the surface of the FRC or by performing silane and bonding treatment. In the Estenia/BR-100 combination, when the adhesive strength was low, the fracture load of the laminate specimens was also low. However, the difference in fracture load was not as large as that seen in adhesive strength. The fracture load of each laminate specimen was significantly lower after thermocycling. The adhesive strength between the FRCs and veneering composite had an effect on the fracture load of the combination.


Th22, but not Th17 might be a good index to predict the tissue involvement of systemic lupus erythematosus.
T-helper (Th) cells abnormalities are considered to be associated with the pathogenesis of Systemic lupus erythematosus (SLE). Recently, The Th22 cells have been identified and implicated in the pathogenesis of autoimmune diseases such as Rheumatoid arthritis (RA), although therir role in Systemic lupus erythematosus (SLE) remains unclear. The present study intends to investigate their roles in SLE. Clinical data were collected in 65 SLE patients and 30 healthy controls. The patients were divided into active and inactive groups. CD4(+)IFN-γ(-)IL-17(-)IL-22(+)Tcells (Th22 cells),CD4(+) IFN-γ(-)IL-22(-)IL-17(+)T cells (Th17 cells),and CD4(+) IFN-γ(+) (Th1 cells) were assayed by flow cytometry. Serum interleukin-22 (IL-22) and IL-17 levels were measured by enzyme-linked immunosorbent assay. The main observation focused on increased Th22 cells in patients with sole lupus skin disease and decreased Th22 cells in patients with sole lupus nephritis. Likewise, concentrations of serum IL-22 were increased in patients with sole lupus skin disease, and decreased in patients with sole lupus nephritis. Additionally, there was a positive correlation between the percentage of Th22 cells and IL-22 production. The percentage of Th17 cells or concentration of serum IL-17 correlated positively with Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Th22 seems to be a more significant index to predict the tissue involvement of SLE than Th17, although Th17 may play a role in the activity of SLE.


Acute Cholangitis Secondary to a Clogged Biliary Stent: A Review on the Cause of Clogging and the Appropriate Time of Replacement.
Choledocholithiasis is preferably treated by endoscopic retrograde cholangiopancreatography (ERCP) with sphincterotomy and stone removal, to reduce the risk for acute cholangitis. Frequently, patients who are ill, surgically unfit, or unable to undergo stone extraction during the index procedure underwent antibacterial treatment and temporary biliary stenting via ERCP to prevent biliary sepsis and septic shock. After a period of convalescence, a repeat ERCP is scheduled to clear the bile duct and remove the stent, followed by laparoscopic cholecystectomy to complete the treatment circuit. Cholangitis may often recur in patients with an indwelling biliary stent while waiting for definitive treatment. Here, we present a case of a 42-year-old female with choledocholithiasis who developed moderate acute cholangitis 5 months after ERCP and insertion of a biliary plastic stent. She was provisionally diagnosed with obstructive jaundice with concurrent acute cholecystitis. Through intravenous antibacterial therapy, stent exchange, and an interval open cholecystectomy, she had fully recovered. We also discuss the underlying mechanism of stent blockage and the optimal interval for stent exchange after temporary placement for benign cases. Understanding the pathophysiology of stent clogging and recognizing the optimal interval for stent replacement may help reduce the risk of stent clogging and potentially fatal acute cholangitis.


BH3-mimetics- and cisplatin-induced cell death proceeds through different pathways depending on the availability of death-related cellular components.
Owing to their important function in regulating cell death, pharmacological inhibition of Bcl-2 proteins by dubbed BH3-mimetics is a promising strategy for apoptosis induction or sensitization to chemotherapy. However, the role of Apaf-1, the main protein constituent of the apoptosome, in the process has yet not been analyzed. Furthermore as new chemotherapeutics develop, the possible chemotherapy-induced toxicity to rapidly dividing normal cells, especially sensitive differentiated cells, has to be considered. Such undesirable effects would probably be ameliorated by selectively and locally inhibiting apoptosis in defined sensitive cells. Mouse embryonic fibroblasts (MEFS) from Apaf-1 knock out mouse (MEFS KO Apaf-1) and Bax/Bak double KO (MEFS KO Bax/Bak), MEFS from wild-type mouse (MEFS wt) and human cervix adenocarcinoma (HeLa) cells were used to comparatively investigate the signaling cell death-induced pathways of BH3-mimetics, like ABT737 and GX15-070, with DNA damage-inducing agent cisplatin (cis-diammineplatinum(II) dichloride, CDDP). The study was performed in the absence or presence of apoptosis inhibitors namely, caspase inhibitors or apoptosome inhibitors. BH3-mimetic ABT737 required of Apaf-1 to exert its apoptosis-inducing effect. In contrast, BH3-mimetic GX15-070 and DNA damage-inducing CDDP induced cell death in the absence of both Bax/Bak and Apaf-1. GX15-070 induced autophagy-based cell death in all the cell lines analyzed. MEFS wt cells were protected from the cytotoxic effects of ABT737 and CDDP by chemical inhibition of the apoptosome through QM31, but not by using general caspase inhibitors. BH3-mimetic ABT737 not only requires Bax/Bak to exert its apoptosis-inducing effect, but also Apaf-1, while GX15-070 and CDDP induce different modalities of cell death in the absence of Bax/Bak or Apaf-1. Inclusion of specific Apaf-1 inhibitors in topical and well-localized administrations, but not in systemic ones, to avoid interferences with chemotherapeutics would be of interest to prevent chemotherapeutic-induced unwanted cell death which could improve cancer patient care.


[Effects of synoptic type on surface ozone pollution in Beijing].
Ozone (O), influenced by meteorological factors, is a primary gaseous photochemical pollutant during summer to fall in Beijing' s urban ambient. Continuous monitoring during July to September in 2008 was carried out at four sites in Beijing. Analyzed with synoptic type, the results show that the ratios of pre-low cylonic (mainly Mongolia cyclone) and pre-high anticylonic to total weather conditions are about 42% and 20%, illustrating the high-and low-ozone episodes, respectively. At the pre-low cylonic conditions, high temperature, low humidity, mountain and valley winds caused by local circulation induce average hourly maximum ozone concentration (volume fraction) up to 102.2 x 10(-9), negative correlated with atmospheric pressure with a slope of -3.4 x 10(-9) Pa(-1). The time of mountain wind changed to valley wind dominates the diurnal time of maximum ozone, generally around 14:00. At the pre-high anticylonic conditions, low temperature, high humidity and systematic north wind induce average hourly maximum ozone concentration (volume fraction) only 49.3 x 10(-9), the diurnal time of maximum ozone is deferred by continuous north wind till about 16:00. The consistency of photochemical pollution in Beijing region shows that good correlation exists between synoptic type and ozone concentration. Therefore, getting an eye on the structure and evolution of synoptic type is of great significances for forecasting the photochemical pollution.


Dose and image quality optimization in neonatal radiography.
In a special care baby unit, neonates, mainly premature, encounter serious to life-threatening diseases, the timely diagnosis and treatment of which may require a large number of radiographs. Increased neonatal radiosensitivity and longer life expectancy increase the risk of radiation-induced cancer, which emphasizes the importance of minimizing dose while maintaining clinically satisfactory image quality. An optimization study on radiation dose and image quality in neonatal radiography is presented. Neonates were categorized into four groups depending on birthweight. For a total of 378 chest and chest-abdomen radiographs, exposure parameters were recorded. Entrance surface dose (ESD) was estimated and dose-area product (DAP) was measured. Image quality evaluation was performed by two observers and was based on the visibility of certain anatomical features and catheters placed during treatment using a five-grade scale. ESD values increased with neonatal weight and demonstrated wide variation (16.4-76.9 microGy, mean 38.2 microGy). A wide variation was also observed in DAP values (1.2-15.0 mGycm2, mean 7.2 mGycm2). Image quality evaluation revealed the feasibility of achieving a diagnostically satisfactory image (score >70%) using both low and high tube voltage techniques, with the latter resulting in reduced ESDs. The majority of estimated ESDs are in accordance with the reference level of 50 microGy recommended by the National Radiological Protection Board for neonatal radiography. The results suggest that the use of high tube voltage techniques could result in further reductions in neonatal dose, without image quality degradation, underlying the requirement for establishing standard examination protocols for neonatal radiography with respect to neonatal weight.


New formula for quantification of pleural effusions from computed tomography.
The authors develop a method to accurately and easily estimate the volume of pleural effusions with computed tomography (CT). In 15 patients with either simple or loculated pleural effusions (14 right-sided and 11 left-sided), routine helical CT examinations of the thorax were obtained. Two experienced radiologists visually estimated the volume of the effusions. Three-dimensional reconstructions of the pleural effusions were performed from the helical CT examinations, and the volumes of the effusions were calculated. Effusion volumes were also estimated using the formula d2 x l (d = greatest depth of the effusion on a single CT image, l = greatest length of the effusion). The computer calculated the estimated volumes and they were then statistically compared. The coefficient of correlation between the estimation by measurement and calculated volumes of all effusions was 0.908 (p<0.0001) for the right side, and 0.849 for the left side (p<0.002). Excluding the loculated effusions, the coefficient of correlation was 0.969 for the right side and 0.949 for the left side (p<.001). The volume estimation by visual inspection was also correlated to the calculated volumes (0.84 on the left and 0.85 on the right, p<0.008), but resulted in a consistent overestimation by 300 ml to 500 ml. Although pleural effusion volumes can be estimated by visual inspection with good correlation, some overestimation is consistently seen. Use of the formula d2 x l readily enables estimation of pleural effusion volume from CT, from two simple measurements. This formula-based method of volume estimation provides an accurate and easily measured means of estimation that is readily obtained from routine CT of the chest.


Common concerns among hospital-based dialysis providers.
Studies by Alan Collins, MD, and the U.S. Renal Data System have demonstrated that a proper balance of good dialysis therapies, including appropriate use of medications, leads to improved patient outcomes, including lower hospitalization rates and mortality. Additionally, this reduction in hospitalization and increase in patient longevity will improve the financial performance of both hospital-based and freestanding dialysis providers. Good care for the patients is of financial benefit to the dialysis program for both hospital-based and freestanding dialysis facilities. This article highlights many of the challenges facing hospital-based dialyis providers, but it only scratches the surface. Hospital-based programs, while in the minority of dialysis providers, need to incorporate many of the lessons that the freestanding providers have learned. These lessons are not just modification of staffing models and the monitoring of productivity standards; the focus is on improving quality of care and capturing all of the revenue the program is entitled to receive. For hospital-based dialysis programs, improving patient outcomes also improves financial performance.


Vasorelaxant Effects of the Vitex Agnus-Castus Extract.
This study was undertaken to describe and characterize the relaxing effects of the medicinal plant Vitex agnus-castus (VAC) extract on isolated rabbit arterial rings. The VAC extracts (VACE) were extracted with ethanol and tested in aorta rings (3-4 mm) of rabbits suspended in an organ bath (Krebs, 37°C, 95% O2/5% CO2) under a resting tension of 1 g to record isometric contractions. After the stabilization period (1-2 hours), contractions were induced by the addition of phenylephrine (0.5 μM) or high KCl (80 mM) and VACE was added on the plateau of the contractions. Experiments were performed to determine the effects and to get insights into the potential mechanism involved in VACE-induced relaxations. The cumulative addition of VACE (0.15-0.75 mg/mL) relaxed, in a concentration-dependent manner, the rabbit aorta rings precontracted either with phenylephrine- or with high KCl thus suggesting calcium channel blocking activities. The VACE effect appeared to be endothelium-dependent. The preincubation with L-NAME (the inhibitor of nitric oxide synthases (NOS)), ODQ (the selective inhibitor of guanylyl cyclase), and indomethacin (the cyclooxygenase inhibitor), downregulated VACE-induced relaxation of aorta rings precontracted with phenylephrine, whereas the bradykinin (stimulator of NOS) and zaprinast (phosphodiesterase inhibitor) further upregulated relaxant effects induced by VACE. These results revealed that the aorta relaxation effect of VACE was mainly endothelium-dependent and mediated by NO/cGMP and prostaglandins synthesis. This vasodilator effect of VACE may be useful to treat cardiovascular disorders, including hypertensive diseases.


General Effect of van der Waals Interactions on the Stability of Alkoxy Intermediates on Metal Surfaces.
The critical role of noncovalent van der Waals (vdW) interactions in determining the relative thermodynamic stability of alkoxy intermediates has been demonstrated for the Cu(110) surface using a combination of experiment and theory. The results may be significant for the selectivity control of copper-based reactions of alcohols. Previous examination of this effect on Au(110) was also extended to include higher molecular weight alcohols; on Cu(110) and Au(110) the hierarchy for the strength of binding of the alkoxys was found to be the same within experimental accuracy, with alkoxy species of greater chain length being more stable. The equilibrium constants governing the competition of alcohol pairs for binding sites of the alkoxys are also similar on the two surfaces. These results reveal the generality of such vdW effects. This work expands the understanding of the role of vdW interactions on the binding efficacy of key reactive intermediates on metal surfaces, a key factor in the rational design of complex and selective catalytic processes.


Sequencing of two transgenic early-flowering poplar lines confirmed vector-free single-locus T-DNA integration.
Next-generation sequencing (NGS) approaches are attractive alternatives to the PCR-based characterisation of genetically modified plants for safety assessment and labelling since NGS is highly sensitive to the detection of T-DNA inserts as well as vector backbone sequences in transgenic plants. In this study, two independent transgenic male Populus tremula lines, T193-2 and T195-1, both carrying the FLOWERING LOCUS T gene from Arabidopsis thaliana under control of a heat-inducible promoter (pHSP::AtFT) and the non-transgenic control clone W52, were further characterised by NGS and third-generation sequencing. The results support previous findings that the T-DNA was hemizygously inserted in one genomic locus of each line. However, the T-DNA insertions consist of conglomerations of one or two T-DNA copies together with a small T-DNA fragment without AtFT parts. Based on NGS data, no additional T-DNA splinters or vector backbone sequences could be identified in the genome of the two transgenic lines. Seedlings derived from crosses between the pHSP::AtFT transgenic male parents and female wild type plants are therefore expected to be T-DNA splinter or vector backbone free. Thus, PCR analyses amplifying a partial T-DNA fragment with AtFT-specific primers are sufficient to determine whether the seedlings are transgenic or not. An analysis of 72 second generation-seedlings clearly showed that about 50% of them still reveal the presence of the T-DNA, confirming data already published. To prove if unanticipated genomic changes were induced by T-DNA integration, extended future studies using long-range sequencing technologies are required once a suitable chromosome-level P. tremula reference genome sequence is available.


Frequent methamphetamine use is associated with primary non-nucleoside reverse transcriptase inhibitor resistance.
We determined whether methamphetamine use is associated with the increased prevalence of primary HIV drug resistance among a cohort of men who have sex with men recently infected with HIV. In multivariate analysis, we found that frequent methamphetamine use was strongly associated with primary non-nucleoside reverse transcriptase inhibitor resistance, but not with protease inhibitor or nucleoside reverse transcriptase inhibitor resistance. We postulate that this association may be caused by methamphetamine-associated treatment interruptions among source partners.


A new strategy for seamless gene editing and marker recycling in Saccharomyces cerevisiae using lethal effect of Cwp1.
Technologies development for seamless gene editing and marker recycling has allowed frequent genomic engineering in Saccharomyces cerevisiae for desired laboratory strains and cell factory. Alternative new approaches are still required for complicated scenarios. In this study, we report that inducible overexpression of cell wall protein 1 (Cwp1) by galactose addition confers yeast cells a robust growth inhibition. Direct repeats flanking the Gal-CWP1:selectable marker cassette allow for its homology recombination excision and counter selection upon galactose addition, therefore enable seamless gene editing and marker recycling. We used this strategy and efficiently generated scarless Ade8 deletion mutants. Our results highlight the utility of lethal effect of Cwp1 overexpression a new counter selection strategy and a simple and efficient method for seamless gene editing and marker recycling in S. cerevisiae and potentially other fungi.


Reliability of 31P-magnetic resonance spectroscopy during an exhaustive incremental exercise test in children.
This study examined the reliability of (31)P-magnetic resonance spectroscopy (MRS) to measure parameters of muscle metabolic function in children. On separate days, 14 children (7 boys and 7 girls) completed three knee-extensor incremental tests to exhaustion inside a whole-body scanner (1.5 T, Phillips). The dynamic changes in the ratio of inorganic phosphate to phosphocreatine (Pi/PCr) and intracellular muscle pH were resolved every 30 s. Using plots of Pi/PCr and pH against power output (W), intracellular thresholds (ITs) for each variable were determined using both subjective and objective procedures. The IT(Pi/PCr) and IT(pH) were observed subjectively in 93 and 81% of their respective plots, whereas the objective method identified the IT(Pi/PCr) in 88% of the plots. The IT(pH) was undetectable using the objective method. End exercise (END) END(Pi/PCr), END(pH), IT(Pi/PCr) and IT(pH) were examined using typical error statistics expressed as a % coefficient of variation (CV) across all three exercise tests. The CVs for the power output at the subjectively determined IT(Pi/PCr) and IT(pH) were 10.6 and 10.3%, respectively. Objective identification of the IT(Pi/PCr) had a CV of 16.3%. CVs for END(pH) and END(Pi/PCr) were 0.9 and 50.0%, respectively. MRS provides a valuable window into metabolic changes during exercise in children. During knee-extensor exercise to exhaustion, END(pH) and the subjectively determined IT(Pi/PCr) and IT(pH) demonstrate good reliability and thus stable measures for the future study of developmental metabolism. However, the objectively determined IT(Pi/PCr) and END(Pi/PCr) displayed poor reliability.


Clinical evaluation of the 350-kV chest radiography system.
A chest radiography unit employing 350 kV was evaluated and found to be superior to conventional low-kV systems. Visualization of air/soft-tissue boundaries, mediastinal structures, the retrocardiac area, and lung apices was improved. Other benefits of the high-kV system include reproducibility, reliability, rapid installation, and economy of operation.


Electroacupuncture and postoperative pain.
We studied 20 otherwise healthy women undergoing lower abdominal surgery. Immediately after wound closure, while still anaesthetized, they received either electroacupuncture (EA) or no further treatment. They were allowed pethidine for postoperative analgesia by patient-controlled infusion pump. Signs of postoperative distress (pain, nausea, drowsiness) were evaluated after 2 and 6 h by visual analogue scale scores. The group receiving EA consumed half the quantity of pethidine as that used used by the no treatment group. Two patients in the EA group had no postoperative analgesia in the first 2 h. There was no difference in the assessments of postoperative distress between groups. No patient was aware of having received EA or not.


Cultivation of Green Microalgae in Bubble Column Photobioreactors and an Assay for Neutral Lipids.
There is significant interest in the study of microalgae for engineering applications such as the production of biofuels, high value products, and for the treatment of wastes. As most new research efforts begin at laboratory scale, there is a need for cost-effective methods for culturing microalgae in a reproducible manner. Here, we communicate an effective approach to culture microalgae in laboratory-scale photobioreactors, and to measure the growth and neutral lipid content of that algae. Instructions are also included on how to set up the photobioreactor system. Although the example organisms are species of Chlorella and Auxenochlorella, this system can be adapted to cultivate a wide range of microalgae, including co-cultures of algae with non-algae species. Stock cultures are first grown in bottles to produce inoculum for the photobioreactor system. Algae inoculum is concentrated and transferred to photobioreactors for cultivation in batch mode. Samples are collected daily for the optical density readings. At the end of the batch culture, cells are harvested by centrifuge, washed, and freeze dried to obtain a final dry weight concentration. The final dry weight concentration is used to create a correlation between the optical density and the dry weight concentration. A modified Folch method is subsequently used to extract total lipids from the freeze-dried biomass and the extract is assayed for its neutral lipid content using a microplate assay. This assay has been published previously but protocol steps were included here to highlight critical steps in the procedure where errors frequently occur. The bioreactor system described here fills a niche between simple flask cultivation and fully-controlled commercial bioreactors. Even with only 3-4 biological replicates per treatment, our approach to culturing algae leads to tight standard deviations in the growth and lipid assays.