Neuroprotective potential of plant derived parenchymal stem cells extract on environmental and genetic models of Parkinson disease through attenuation of oxidative stress and neuroinflammation.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by both motor and non-motor features. The current treatment regimen for PD are dopamine enhancers which have been reported to worsen the disease prognosis after long term treatment, thus, the need for better treatment options. This study sought to investigate the protective action of Double Stem Cell® (DSC), a blend of stem cells extracts from Swiss apples (Malus Domestica) and Burgundy grapes (Vitis vinifera) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinsonism in mice and genetic model of PD in Drosophila melanogaster. Male albino mice were pretreated with MPTP (4 × 20 mg/kg, i.p., two hourly in 8 h), twelve hours before administration of DSC (8, 40, or 200 mg/kg, p.o.). Thereafter, behavioural, biochemical and immunohistochemical assays were carried out. The impact of vehicle or DSC supplementation on α-synuclein aggregation was evaluated in Drosophila melanogaster using the UAS-Gal4 system, female DDC-Gal4 flies were crossed with male UAS-α-synuclein, the progenies were examined for fecundity, locomotion, memory, and lifespan. MPTP-induced motor deficits in open field test (OFT), working memory impairment (Y-maze test (YMT)) and muscle incoordination (rotarod test) were ameliorated by DSC (8, 40 or 200 mg/kg) through dose-dependent and significant improvements in motor, cognitive and motor coordination. Moreso, MPTP exposure caused significant increase in lipid peroxidation and decrease in antioxidant enzymes activities (glutathione, catalase and superoxide dismutase) in the midbrain which were attenuated by DSC. MPTP-induced expression of microglia (iba-1), astrocytes (glia fibrillary acidic protein; GFAP) as well as degeneration of dopamine neurons (tyrosine hydroxylase positive neurons) in the substantia nigra (SN) were reversed by DSC. Supplementation of flies feed with graded concentration of DSC (0.8, 4 or 20 mg/ml) did not affect fecundity but improved climbing activity and lifespan. Findings from this study showed that Double Stem Cell improved motor and cognitive functions in both mice and Drosophila through attenuation of neurotoxin-induced oxidative stress and neuroinflammation.


[Similarity of the HL-A antigens in parents of patients with acute leukemia or aplastic anemia].
The frequency of common HLA-A and -B antigens was determined in 30 couples with a child suffering from acute leukemia (AL), 34 couples with a child suffering from aplastic anemia (AA) and 58 random couples with healthy children. Increased frequency of couples sharing at least two common antigens was observed in parents of both AL and AA children.


Polymorphisms in the promoter region of the alpha1A-adrenoceptor gene are associated with schizophrenia/schizoaffective disorder in a Spanish isolate population.
Animal models have implicated the alpha(1)-adrenergic subtypes in cognitive functions relevant to schizophrenia, but no consensus exists with regard to the status of noradrenergic receptor populations in psychiatric patients. We focused on one alpha(1)-adrenergic subtype, the alpha(1A)-adrenergic receptor, and proposed that genetic variants within the regulatory region of this gene (ADRA1A) alter the expression of this receptor, influencing susceptibility toward schizophrenia. This study examined this proposal by testing the hypothesis that single nucleotide polymorphisms (SNPs) in the promoter region of the alpha(1A)-adrenergic gene were associated with schizophrenia by performing case-control association analysis on SNPs found in a 5' upstream region, which included the putative promoter region and 5' untranslated region. Our sample consisted of 103 schizophrenia and 14 schizoaffective disorder patients and 176 control subjects. All recruits were from a Spanish population isolate of Basque origin that is characterized by low heterogeneity, which was selected with the intent that it might facilitate the identification of disease-related polymorphisms. A total of eight SNPs (-9625 G/A, -7255 A/G, -6274 C/T, -4884 A/G, -4155 C/G, -2760 A/C, -1873 G/A, and -563 C/T) were confirmed at a rare allele frequency of >5%. Association with schizophrenia and schizoaffective disorder was found for the -563 C/T SNP (p = .0005 for allele and p = .007 for genotype, Bonferroni corrected) and -9625 G/A SNP (p = .02 for allele and p = .03 for genotype, Bonferroni corrected). Significant differences in the 54 haplotypes formed by these eight SNPs were also found between patients and control subjects (p = .008, Bonferroni corrected). Because of the strength of these results and the location of these SNPs in the regulatory region of this gene, functional studies investigating the possible influence of these SNPs on receptor expression levels in schizophrenia are warranted.


p38MAPK Signaling Enhances Glycolysis Through the Up-Regulation of the Glucose Transporter GLUT-4 in Gastric Cancer Cells.
Previous studies have shown that p38MAPK is involved in gastric cancer, yet the underlying mechanism remains unclear. q-PCR, Western blot and immunohistochemistry were used to explore the expression of PP2A and the phosphorylation of p38MAPK in gastric cancer tissues and normal gastric tissues. Activated p38MAPK in the gastric cancer cell line MKN45 using activator, then q-PCR, glucose uptake assay and colony formation assay were performed to determine whether p38MAPK promotes gastric cancer through the enhancement of glycolysis. After transfection of p38MAPK dominant negative mutation (p38DN) into MKN45 cells or MKN45 cells treated with an inhibitor of p38MAPK, Western blot was performed to detect the expression of GLUT-4. The knock down of MEF2α in MKN45 cells by siRNA was followed by Western blot and luciferase reporter assay to investigate the underlying mechanism of the role of p38MAPK in the promotion of gastric cancer. Finally, q-PCR, Western blot and immunohistochemistry were performed to examine GLUT-4 expression in gastric cancer tissues and normal gastric tissues. We found that p38MAPK activation significantly increases GLUT-4 expression and promotes glucose uptake and cell growth in gastric cancer cells. Inhibition of p38MAPK abrogates the up-regulation of GLUT-4. MEF2α knockdown abolishes p38MAPK-mediated GLUT-4 up-regulation. PP2A, an inhibitor of p38MAPK, is down-regulated in gastric cancer tissues, which might contribute to the activation of p38MAPK. Our data indicate that the abnormal activation of p38MAPK promotes glycolysis within gastric cancer cells through the upregulation of GLUT-4 in a MEF2a-dependent manner.


BMX activates Wnt/β-catenin signaling pathway to promote cell proliferation and migration in breast cancer.
Breast cancer has become a dangerous killer for the female, which seriously threatened women's life, leading to huge pressures to society. The present study assessed the mechanism underlying the involvement of bone marrow tyrosine kinase on chromosome X (BMX) in breast cancer development. The expression of BMX was examined by qPCR and immunohistochemistry. The effect of BMX on cell proliferation and migration was detected by Clone formation assay and Transwell assay. In vitro study, the correlation of BMX with Wnt/β-catenin pathway was explored by western blot and TOP/FOP flash assay. In the present study, we found that BMX was up-regulated in breast cancer, which was associated with the tumor differentiation and TNM stage. Oncogenic BMX enhanced the ability of breast cancer cell proliferation and migration. Furthermore, BMX could up-regulate the protein expression levels of p-β-catenin (Y142), p-β-catenin(Y654) and inhibit the expression level of p-β-catenin (S33/37), thus activating Wnt/β-catenin pathway in MCF-7 and MDA-MB-231 cells. In addition, we revealed that BMX promoted GSK3β phosphorylation, which suppressed the degradation of β-catenin. In this study, we identified that BMX-activated Wnt/β-catenin signaling pathway, playing an oncogenic role in breast cancer, suggesting that BMX could become a potential treatment target of breast cancer.


Toxic epidermal necrolysis complicating a burn: a case report.
The case of a burned patient, whose fluid management was complicated by the development of toxic epidermal necrolysis, a condition which itself may mimic a burn, is presented. The possible causes, together with the diagnostic and therapeutic problems posed by this combination of ailments, are discussed.


Natural autoantibodies in nude and normal outbred (Swiss) and inbred (BALB/c) mice.
Spleen cells from adult unprimed outbred (Swiss) and inbred (BALB/c) mice, either normal (no) or athymic-nude (nu) as well as spleen cells from Swiss nude mice bearing two different human tumors (BUR and PINQ), were fused with the mouse non-secreting myeloma cell line P3X63 Ag8-653. The supernatants of immunoglobulin secreting hybrids, all containing IgM, were screened for antibody activity against macromolecular antigens (autologous: actin, tubulin, myosin, dsDNA) and haptens (TNP, NP, NIP and NBrP). Furthermore, their idiotypic determinants were analyzed using a rabbit anti-idiotype which recognizes a major cross-reactive idiotype (IdD23) of BALB/c natural polyreactive autoantibodies. In all the mice studied, we identified: (1) hybrids reacting strongly with one or more haptens (10.7 to 37.8%) and (2) hybrids secreting natural monoclonal autoantibodies (NMoAb) with broad reactivities (polyreactive and/or oligoreactive) against autoantigens and/or haptens (11.4 to 26.8%). The results indicate that: (1) cells secreting natural autoantibodies with broad reactivities exist in both normal and nude mice, independently of the genetic background (inbred/outbred) of the mouse. However, in nude mice, the natural autoantibodies exhibit a more restricted pattern of reactivity (oligoreactive) compared to those of normal mice, and do not express the common idiotype IdD23 of natural polyreactive autoantibodies. (2) Tumors grafted into nude mice seem to induce the expression of polyreactive autoantibodies bearing the IdD23.


Role of the hypothalamus in ghrelin effects on reproduction: sperm function and sexual behavior in male mice.
Ghrelin signals to the hypothalamus inhibit reproduction during times of food scarcity. In this study, we demonstrate that ghrelin impairs sperm quality in male mice. Ghrelin (GHRL) is an orexigenic peptide that has been investigated as one of the signals responsible for the reproductive performance of mammals under fluctuating metabolic conditions. Central GHRL administration impairs spermatogenesis in mice by regulating the hypothalamic-pituitary-gonadal axis function. In the present study, the hypothalamus role as a mediator of GHRL effects on sperm fertilizing capacity and male sexual behavior was evaluated. After 42 days of hypothalamic GHRL infusion or artificial cerebrospinal fluid, in vitro and in vivo sperm fertilizing capacity, testicular α-tubulin, speriolin gene expression and spermatic α-tubulin protein were evaluated. Hypothalamic expression of genes Kiss1, Gpr54 and Gnrh was also studied. The second group of animals was infused with one time only GHRL or artificial cerebrospinal fluid into the hypothalamus to evaluate the effects on sexual behavior. Results demonstrated that chronic GHRL administration to male mice significantly increased the percentages of pre-implantation embryo loss and the number of post-implantation embryo loss. In relation to the gene expression, our results show a relative decrease of Kiss1, Gpr54 and Spatc1. Although no significant differences were observed in the quantitative expression of α-tubulin protein, qualitative changes in its expression pattern were observed. In addition, a dual effect on sexual behavior was observed: 40% of the treated animals showed a significant reduction in the number of mounts and intromissions, while a 60% showed a significant decrease in ejaculation latency vs control animals. In conclusion, our results provide evidence that central GHRL administration possibly induces failure in embryo development and/or implantation in the females mated with treated males, possibly because of a negative effect in the α-tubulin pattern.


Diagnosis of invasive candidiasis in the ICU.
Invasive candidiasis ranges from 5 to 10 cases per 1,000 ICU admissions and represents 5% to 10% of all ICU-acquired infections, with an overall mortality comparable to that of severe sepsis/septic shock. A large majority of them are due to Candida albicans, but the proportion of strains with decreased sensitivity or resistance to fluconazole is increasingly reported. A high proportion of ICU patients become colonized, but only 5% to 30% of them develop an invasive infection. Progressive colonization and major abdominal surgery are common risk factors, but invasive candidiasis is difficult to predict and early diagnosis remains a major challenge. Indeed, blood cultures are positive in a minority of cases and often late in the course of infection. New nonculture-based laboratory techniques may contribute to early diagnosis and management of invasive candidiasis. Both serologic (mannan, antimannan, and betaglucan) and molecular (Candida-specific PCR in blood and serum) have been applied as serial screening procedures in high-risk patients. However, although reasonably sensitive and specific, these techniques are largely investigational and their clinical usefulness remains to be established. Identification of patients susceptible to benefit from empirical antifungal treatment remains challenging, but it is mandatory to avoid antifungal overuse in critically ill patients. Growing evidence suggests that monitoring the dynamic of Candida colonization in surgical patients and prediction rules based on combined risk factors may be used to identify ICU patients at high risk of invasive candidiasis susceptible to benefit from prophylaxis or preemptive antifungal treatment.


SCD1 inhibits HBV replication by regulating autophagy under high lipid conditions.
Chronic hepatitis B virus (HBV) infection remains a significant public health concern worldwide. Several metabolic processes regulate HBV DNA replication, including autophagy and lipid metabolism. In this study, we clarified the effect of lipids on HBV replication and elucidated possible mechanisms. We discovered that lipid metabolic gene expression levels were negatively correlated with the HBV DNA in plasma. Our data showed that fatty acid stimulation significantly reduced HBV DNA, hepatitis B surface antigen (HBsAg), and hepatitis B e antigen (HBeAg) levels in HepG2.2.15 cells, which are human hepatoma cell cultures transfected with HBV DNA. The Stearoyl coenzyme A desaturase 1 (SCD1)-autophagy pathway has also been implicated in inhibiting HBV replication by fatty acids stimulation. SCD1 knockdown deregulates the inhibitory effect of fatty acids on HBV by enhancing autophagy. When 3 methyladenine (3MA) was added, the inhibitory effects of specific autophagy inhibitors eliminated the positive effects of SCD1 knockdown on HBV replication. Our results indicate that SCD1 participates in the regulation of inhibition of HBV replication by fatty acids stimulation through regulating autophagy.


A numerical simulation study of the hydrodynamic effects caused by morphological changes in the Guadalquivir River Estuary.
This study presents an analysis of the impacts of the changes in bottom depth along the Guadalquivir Estuary on tidal dynamics. A realistic non-linear 1D numerical model, incorporating changes in both breadth and bottom depth, was employed to investigate the involved effects. The findings reveal a significant amplification of the M2 tidal wave towards the upper region of the Estuary, resulting from the gradual deepening caused by multiple dredging operations. The Estuary exhibits a pronounced tendency towards resonance, which is further enhanced by its deepening, resulting in reduced bottom friction and a smaller decrease in tidal wave amplitude as it propagates through the Estuary. The alterations in depth, particularly in breadth, along the Estuary play a crucial role in determining the magnitude of the resonant response of the M2 tidal wave.


Optimized precision of inversion-recovery T1 measurements for constrained scan time.
Optimization of inversion-recovery (IR) measurements of T1 relaxation times has focused on the efficiency of T1 estimation (precision per unit time), without explicit consideration of the total experimental time. With the modified fast IR method, the repetition time (TR) is constant and each inversion time (TI) must be less than TR. For a fixed total experimental time in the absence of signal averaging, TR and the longest TI time must decrease as more TI times are added. This article investigates the number of TI times and their values to optimize the precision of T1 measurements when time constraints preclude signal averaging and limit the largest TI that can be acquired. If the longest TI is less than approximately three times the longest T1 of interest, then precision is optimum with only four TI times. Suggestions are given for selecting the optimum TI times for 1-16-fold ranges of T1 times relative to a given maximum T1.


Optic disk cupping and pallor measurements of patients with a disk hemorrhage.
We evaluated whether the rate of occurrence of glaucomatous disk damage in eyes with an optic disk hemorrhage differed from the rate of occurrence in control eyes. We compared 24 patients with a disk hemorrhage to 24 control patients, who had been matched for diagnosis, disk appearance, and visual fields. Cup volume and cup-to-disk area ratio of the optic disk were measured using the stereophotogrammetric technique, and area of pallor was measured using computerized image analysis. We found that diabetes increased the probability that a patient would have a disk hemorrhage, and that a hemorrhage was associated with progression of glaucomatous disk changes in half of the cases during the mean 3.1-year follow-up period. However, the mean rate of glaucomatous disk damage in patients with a hemorrhage was not different from matched control patients. A disk hemorrhage is a sign of later disk damage, but it does not alter the rate of glaucomatous disk progression.


Intraarterial digital subtraction angiography vs conventional angiography of abdominal diseases.
Seventy-five patients with various abdominal diseases were studied by intraarterial digital subtraction angiography (IA DSA) during conventional abdominal angiographic examinations. The image quality, tumor stain, and arterial encasements were evaluated and diameters of vessels visualized by IA DSA were measured. The vessels greater than 2.0 mm or more in diameter were equally well demonstrated on IA DSA. IA DSA demonstrated tumor stains and portal system to good advantage. Real time imaging was also advantageous during an embolization procedure.


Function and distribution of three rat 5-hydroxytryptamine7 (5-HT7) receptor isoforms produced by alternative splicing.
Serotonin (5-HT7) receptor pre-mRNA is alternatively spliced in rat tissue to produce three isoforms, 5-HT(7a), 5-HT(7b) and 5-HT(7c), which differ in the amino acid sequences of their carboxyl terminal tails. Substantial species differences in structure and expression patterns exist for 5-HT7 isoforms. We have now compared some of the functional characteristics and level of expression for the three rat 5-HT7 receptor isoforms. Recombinant receptor isoforms were expressed in COS-7 cells for examination of [3H]5-HT binding characteristics and in JEG-3 cells to ascertain their ability to stimulate cAMP production. These studies showed that all three isoforms are functionally active and have similar agonist binding characteristics. Distribution of expression of the three rat receptor isoforms were examined in several brain regions and peripheral tissues using RT-PCR and in situ hybridization. The relative proportions of total 5-HT7 receptor message lent by each isoform varied little between these areas. In contrast to what has been observed in human tissue, the 5-HT(7a) isoform predominated in all regions examined, while the 5-HT(7c) isoform revealed a low level of expression (3% of total transcript). In situ hybridization was used to determine if the overall low level of expression of the 5-HT(7c) isoform by RT-PCR could be attributed to a small localized subpopulation of cells expressing high levels 5-HT(7c) message. In situ hybridization results indicate a generalized low level of expression of the 5-HT(7c) isoform throughout the CNS. These data suggest that while all three known 5-HT7 receptor isoforms in the rat are functionally competent, any functionally important differences between the three isoforms are not likely to involve differences in ligand binding or gross differences in adenylate cyclase coupling. However, differences in receptor phosphorylation, regulation or coupling to other effectors or cell trafficking could still exist.


An actin depolymerizing agent 19,20-epoxycytochalasin Q of Xylaria sp. BCC 1067 enhanced antifungal action of azole drugs through ROS-mediated cell death in yeast.
Multidrug resistance is a highly conserved phenomenon among all living organisms and a major veritable public health problem worldwide. Repetitive uses of antibiotics lead to antimicrobial drug resistance. Here, 19,20-epoxycytochalasin Q (ECQ) was isolated from endophytic fungus Xylaria sp. BCC 1067 and, its chemical structure was determined via chromatographic and spectral methods. ECQ displayed an antifungal activity with low MIC50 of 410 and 55 mg/l in the model yeast Saccharomyces cerevisiae wild-type and ScΔpdr5 strains, respectively. ECQ was a new inducer and potential substrate of key multi-drug efflux pumps S. cerevisiae ScPdr5 and Candida albicans CaCdr1. ECQ targeted actin filament, disrupting actin dynamics of yeast cells. ECQ also sensitized the ScΔsrv2 mutant, lacking suppressor of RasVal19. Overexpression of ScPDR5 or CaCDR1 genes prevented aggregation of actin and alleviated antifungal effect of ECQ. Additionally, ECQ induced high accumulation of reactive oxygen species, caused plasma membrane leakage and decreased yeast cell survival. Importantly, a discovery of ECQ implied a cellular connection between multi-drug resistance and actin stability, an important determinant of transporter mediated-drug resistance mechanism. Combination of ECQ and antifungal azoles displayed promising drug synergy against S. cerevisiae strains expressing multi-drug transporters, thereby providing potential solution for antifungal therapy and chemotherapeutic application.


Construction waste as substrate in vertical subsuperficial constructed wetlands treating organic matter, ibuprofenhene, acetaminophen and ethinylestradiol from low-strength synthetic wastewater.
This study aimed to evaluate the removal of chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), total ammonia nitrogen (TAN), total phosphorus (TP), ibuprofen, acetaminophen and ethinylestradiol of synthetic effluent simulating low-strength sewage by sequencing-batch mode constructed wetlands (CWs). To verify the feasibility of using a floating macrophyte in CWs and compare different substrates, three CWs containing light expanded clay aggregates (CWL), expanded clay with porcelain tiles (CWLP) and bricks (CWB) were planted with Pistia stratiotes. The results showed that CWB achieved the highest removals of TKN (78%), TAN (70%) and TP (46%), and CWLP achieved the highest COD removal (75%). LECA favored the removal of ibuprofen (92%, p < 0.05) when compared to bricks (77%), probably by the combination of biodegradation and sorption in the systems. The highest acetaminophen removal (71% to 96%) was observed in CWL, probably via biodegradation, but no significant differences were found between the CWs (p > 0.05). Ethinylestradiol was removed 76% in CWLP and 73% in CWB, both differing statistically from CWL (p < 0.05), demonstrating that brick and the combination of clay with porcelain were better than just clay in this hormone removal. After 188 days of operation, P. stratiotes was able to uptake nitrogen and phosphorus of approximately 0.28 g and 0.25 g in CWL, 0.33 g and 0.21 g CWLP, and 0.22 g and 0.09 g in CWB of, respectively. Adsorption of nitrogen and phosphorus onto the substrates was 0.48 g and 6.84 g in CWL, 0.53 g and 5.69 g in CWLP, and 0.36 g and 10.18 g in CWB, respectively. The findings on this study suggest that adsorption was possible the main process for TP removal onto the evaluated substrates whereas microbial activity was the most probable mechanism for TN removal in the evaluated CW systems.


Evaluation of degradation mechanism of chlorhexidine by means of Density Functional Theory calculations.
Chlorhexidine (CHD), a germicidal drug, has degradation products that can be hemotoxic and carcinogenic. However, there is no consensus in literature about the degradation pathway. In order to shed light on that mechanism, we have employed Density Functional Theory to study reactants, in different protonation states, products and intermediates involved in the different pathways. Based on free energy values comparison and frontier molecular orbital analysis, we have obtained the most stable structures in each protonation state. CHD in saturated form has HOMO localized in one p-chloroaniline, and, due to molecule's symmetry, HOMO-1 has contributions from the other side of the molecule, but mainly from the biguanide portion of the molecule, instead of from the p-chloroaniline. For the saturated form, we have studied two possible degradation pathways, starting from the monoprotonated structure, and three pathways starting from the neutral structure. We found out that the mechanisms proposed in literature, whose pathways lead to p-chloroaniline (PCA) formation in a smaller number of steps, are more likely than the mechanisms with more intermediate steps or pathways that do not predict PCA formation. Also, based on free energy results, we have found that the formation of another sub-product (PBG-AU) is favorable as well.


Clinical Outcomes of Computed Tomography-Based Volumetric Brachytherapy Planning for Cervical Cancer.
A report of clinical outcomes of a computed tomography (CT)-based image guided brachytherapy (IGBT) technique for treatment of cervical cancer. Seventy-six women with International Federation of Gynecology and Obstetrics stage IB to IVA cervical carcinoma diagnosed between 2007 and 2014 were treated with definitive external beam radiation therapy (EBRT) with or without concurrent chemotherapy followed by high-dose-rate (HDR) IGBT. All patients underwent planning CT simulation at each implantation. A high-risk clinical target volume (HRCTV) encompassing any visible tumor and the entire cervix was contoured on the simulation CT. When available, magnetic resonance imaging (MRI) was performed at implantation to assist with tumor delineation. The prescription dose was prescribed to the HRCTV. The median follow-up time was 17 months. Thirteen patients (17%) had an MRI done before brachytherapy, and 16 patients (21%) were treated without MRI guidance. The mean EBRT/IGBT sum 2-Gy equivalent dose (EQD2) delivered to the 90% volume of the HRCTV was 86.3 Gy. The mean maximum EQD2s delivered to 2 cm(3) of the rectum, sigmoid, and bladder were 67.5 Gy, 66.2 Gy, and 75.3 Gy, respectively. The 2-year cumulative incidences of local, locoregional, and distant failure were 5.8% (95% confidence interval [CI]: 1.4%-14.8%), 15.1% (95% CI: 5.4%-29.4%), and 24.3% (95% CI: 12.1%-38.9%), respectively. The 2-year overall and disease-free survival rates were 75% (95% CI, 61%-91%) and 73% (95% CI, 60%-90%), respectively. Twenty-nine patients (38%) experienced grade ≥ 2 acute toxicity, with 5 cases of acute grade 3 toxicity and no grade ≥ 4 toxicities. One patient experienced grade 3 gastrointestinal toxicity. No other late grade ≥ 3 events were observed. This is the largest report to date of CT/MRI-based IGBT for the treatment of cervical cancer. The results are promising, with excellent local control and acceptable toxicity. Further investigation is needed to assess the long-term safety and efficacy of this treatment.


Etiologic factors in long-term respiratory function abnormalities following esophageal atresia repair.
Recurrent respiratory illnesses are frequent in infants following repair of esophageal atresia and functional abnormalities of respiratory and esophageal function are often seen in older children. Recurrent aspiration is a potential cause of these respiratory abnormalities, but a relationship between abnormalities of gastrointestinal and respiratory mechanics has not been adequately investigated. We sought an association between lower esophageal sphincter (LES) incompetence, gastroesophageal reflux (GER), and respiratory function abnormalities in 18 subjects (age 12 to 21 years) following repair of esophageal atresia (Vogt type 111B). In each subject, measurements were made of spirometry, lung volumes assessed by plethysmography, esophageal manometry recorded using a constantly infused fluid-filled trilumen catheter to assess LES pressure and esophageal motility, and esophageal pH monitoring to detect GER. Subjects were grouped according to the presence or absence of a radiologically supported diagnosis of pneumonia in the first 4 years of life. Lung volumes were mildly but significantly decreased in the "pneumonia" group compared with the "nonpneumonia" group. There was no association between abnormalities of respiratory function and abnormal LES pressure or the presence of GER. These data suggest that pneumonia in esophageal atresia infants is associated with mild long-term lung damage. LES dysfunction and GER do not appear to play a major role in this process.


Computer based method for the three-dimensional kinematic analysis of combined posterior cruciate ligament and postero-lateral complex reconstructions on cadaver knees.
The aim of this study is to evaluate the effect of combined posterior cruciate ligament (PCL) and postero-lateral corner (PLC) reconstruction on laxity and three-dimensional kinematics of cadaver knees. We performed anatomical double bundle PCL reconstruction, and functional one bundle 'over-the-bottom' PCL reconstruction combined with one type of PLC reconstruction, running from the postero-lateral tibia to an isometric point near the lateral epicondyle of the femur. Our results showed that combined reconstruction was necessary to restore rotatory laxity. PLC reconstruction, according to the technique described, invariably created a shift towards internal rotation of the kinematic curves, compared to the intact knee.


The topological structure and function of Echinococcus granulosus lactate dehydrogenase, a tegumental transmembrane protein.
Lactate dehydrogenase (LDH), a terminal glycolytic enzyme, is generally considered as a cytosolic protein. We cloned lactate dehydrogenase from Echinococcus granulosus (EgLDH) and predicted it may be a membrane protein with two transmembrane regions through bioinformatics analysis. Intact worm immunofluorescence with antibodies prepared against linear B cell epitopes predicted in the region inside or outside of the membrane demonstrated that EgLDH spans the tegumental membrane twice, with the N terminal and C terminal all outside, just consistent with the putative topological structure. Then, the enzymatic characteristics and kinetic parameters of recombinant EgLDH were surveyed and the results suggested that EgLDH is responsible for catalyzing the reduction of pyruvic acid into lactic acid under physiological conditions. The enzymatic activity of the recombinant protein was inhibited by antibodies directed against the intact protein or against epitopes that contain key residues in the catalytic center or substrate binding sites. EgLDH is a potential target for drugs and vaccines against E. granulosus.


Comparison between a submerged membrane bioreactor and a conventional activated sludge system on treating ammonia-bearing inorganic wastewater.
A submerged membrane bioreactor (SMBR) and a conventional activated sludge system (CAS) were compared in parallel over a period of 210 days on treating synthetic ammonia-bearing inorganic wastewater under similar conditions. Except for a short period of pH control failure, almost complete conversion of NH(4)(+)?N to NO(3)(-)?N was constantly achieved over an NH(4)(+)?N concentration range from 180 to 1300mgl(-1) at a hydraulic retention time (HRT) of 24h in the SMBR, compared to an average conversion ratio of 95.0% in the CAS. Scanning electron micrographs (SEMs) demonstrated the accumulation of extracellular polymeric substances (EPSs) in the SMBR. Ubiquinone-8 (UQ-8), followed by UQ-10, UQ-7 and UQ-9, was the dominant ubiquinone in both the systems. The dominant menaquinone in the SMBR was menaquinone-6 (MK-6), while that in the CAS was MK-7, indicating that some differences existed between the two systems in terms of microbial community structure. Soluble microbial products (SMPs) tended to accumulate, and then biodegrade in SMBR.


Inhibitory effects of pomegranate concentrated solution on the activities of hyaluronidase, tyrosinase, and metalloproteinase.
Synopsis Botanical antioxidants have attracted much attention as useful preventatives of skin damage. Pomegranate is consumed throughout the world for its beneficial health effects, including its antioxidant and anti-inflammatory activities. We investigated whether pomegranate concentrated solution (PCS) could serve as a potential functional cosmetic ingredient that exerts a skin-whitening effect and antiwrinkle activity. To investigate the moisturizing effect of PCS, hyaluronidase activity was examined in human keratinocytes (HaCaT). Elastase and procollagenase activities were assessed in normal human primary dermal fibroblast-neonatal (HDF-N) cells to determine their antiwrinkle effects. Metalloproteinase 1 (MMP-1) activity was also assessed following ultraviolet A (UVA) irradiation. Whitening effects were measured by a tyrosinase inhibition assay and melanin formation test in mouse melanocytes (Melan-a). In addition, histopathological analysis was performed to determine the number of microfolds formed on the epithelial surface, mean epithelial thickness, mean number of inflammatory cells infiltrating the dermis, and collagen fiber-occupied regions within the dermis. Hyaluronan synthesis was significantly increased by PCS in HaCaT cells, while procollagenase and elastase activities were decreased in HDF-N cells. A significant decrease in UVA-induced MMP-1 activity was also observed in PCS-treated HDF-N cells, compared with UVA-exposed cells. PCS effectively reduced melanin production and mushroom tyrosinase activity in Melan-a cells. Moreover, UVB-induced histopathological dermal sclerosis and inflammatory signs were significantly attenuated in PCS-administered mice compared with UVB-exposed mice. Conclusions: Our results suggest that PCS prevents signs of aging, including those related to photoaging. These effects are associated with enhanced hyaluronan synthesis, as well as suppressed elastase, collagenase, MMP-1, and tyrosinase activities and melanin production. UVB-induced photoaging, such as histopathological dermal sclerosis and inflammatory signs, were effectively reduced on the addition of PCS. These results also suggest that skin aging can be prevented and reduced by the antioxidant effects of PCS.


Mast cell maturation is driven via a group III phospholipase A2-prostaglandin D2-DP1 receptor paracrine axis.
Microenvironment-based alterations in phenotypes of mast cells influence the susceptibility to anaphylaxis, yet the mechanisms underlying proper maturation of mast cells toward an anaphylaxis-sensitive phenotype are incompletely understood. Here we report that PLA2G3, a mammalian homolog of anaphylactic bee venom phospholipase A2, regulates this process. PLA2G3 secreted from mast cells is coupled with fibroblastic lipocalin-type PGD2 synthase (L-PGDS) to provide PGD2, which facilitates mast-cell maturation via PGD2 receptor DP1. Mice lacking PLA2G3, L-PGDS or DP1, mast cell-deficient mice reconstituted with PLA2G3-null or DP1-null mast cells, or mast cells cultured with L-PGDS-ablated fibroblasts exhibited impaired maturation and anaphylaxis of mast cells. Thus, we describe a lipid-driven PLA2G3-L-PGDS-DP1 loop that drives mast cell maturation.


Density-dependent flowering phenology, outcrossing, and reproduction in Impatiens capensis.
We investigated the effects of plant density on cleistogamous (CL) and chasmogamous (CH) flowering phenology and seed production in a natural Impatiens capensis population, by censusing individually marked plants at experimentally reduced and natural densities. CL flowering was earlier at natural density. This plastic density response may have resulted from a stress-related threshold for CL flowering; slower growing plants at natural density flowered earlier. Although apparently triggered by slow early growth, early CL flowering also involved an additional cost for later growth rate. In contrast, CH flowering was unrelated to relative growth rate, but apparently required a size threshold. Experimental density reduction resulted in earlier CH flowering and a dramatic increase in the percentage of plants producing CH flowers. Individual CL and CH flowering duration and flower production were greater at reduced density. These density-dependent effects caused differences between treatments in the shape and location of population flowering phenology curves. Moreover, the percentage of CH seeds produced per individual was much higher at reduced density. At natural density total seed production per plant was lower and more hierarchical than at lower density, suggesting that dominance and suppression shape jewelweed fitness distributions.


Sediment pore water distribution coefficients of PCB congeners in enriched black carbon sediment.
More than 2300 sediment pore water distribution coefficients (KPCBi ds) of 93 polychlorinated biphenyls (PCBs) were measured and modeled from sediments from Indiana Harbor and Ship Canal. KPCBi ds were calculated from previously reported bulk sediment values and newly analyzed pore water. PCBs in pore waters were measured using SPME PDMS-fiber and ∑PCB ranged from 41 to 1500 ng L(-1). The resulting KPCBi ds were ~1 log unit lower in comparison to other reported values. A simple model for the KPCBi d consisted of the product of the organic carbon fraction and the octanol-water partition coefficient and provided an excellent prediction for the measured values, with a mean square error of 0.09 ± 0.06. Although black carbon content is very high in these sediments and was expected to play an important role in the distribution of PCBs, no improvement was obtained when a two-carbon model was used.


Application of artificial neural networks for quantitative analysis of image data in chest radiographs for detection of interstitial lung disease.
The authors have developed an automated computeraided diagnostic (CAD) scheme by using artificial neural networks (ANNs) on quantitative analysis of image data. Three separate ANNs were applied for detection of interstitial disease on digitized chest images. The first ANN was trained with horizontal profiles in regions of interest (ROIs) selected from normal and abnormal chest radiographs for distinguishing between normal and abnormal patterns. For training and testing of the second ANN, the vertical output patterns obtained from the 1st ANN were used for each ROI. The output value of the second ANN was used to distinguish between normal and abnormal ROIs with interstitial infiltrates. If the ratio of the number of abnormal ROIs to the total number of all ROIs in a chest image was greater than a specified threshold level, the image was classified as abnormal. In addition, the third ANN was applied to distinguish between normal and abnormal chest images. The combination of the rule-based method and the third ANN also was applied to the classification between normal and abnormal chest images. The performance of the ANNs was evaluated by means of receiver operating characteristic (ROC) analysis. The average Az value (area under the ROC curve) for distinguishing between normal and abnormal cases was 0.976 +/- 0.012 for 100 chest radiographs that were not used in training of ANNs. The results indicate that the ANN trained with image data can learn some statistical properties associated with interstitial infiltrates in chest radiographs.


Iron-catalyzed cross-dehydrogenative coupling esterification of unactive C(sp3)-H bonds with carboxylic acids for the synthesis of α-acyloxy ethers.
An iron-catalyzed oxidative esterification reaction between unactivated C(sp(3))-H bonds from symmetric and asymmetric ethers and carboxylic acids using di-tert-butyl peroxide (DTBP) as the oxidant via a cross dehydrogenative coupling (CDC) reaction was established, which tolerates a wide range of cyclic ether substrates to react with aromatic acids and phenylacetic acid, providing an efficient method for the preparation of α-acyloxy ethers with good to excellent yields. Intermolecular competing kinetic isotope effect (KIE) experiments were also carried out, which indicate that C(sp(3))-H bond cleavage may be the rate-determining step of this CDC reaction.


One-dimensional/two-dimensional hybridization for self-supported binder-free silicon-based lithium ion battery anodes.
A unique silicon-based anode for lithium ion batteries is developed via the facile hybridization of one-dimensional silicon nanowires and two-dimensional graphene sheets. The resulting paper-like film holds advantages highly desirable for not only accommodating the volume change of silicon, but also facilitating the fast transport of electron and lithium ions.


Neurological signs in 23 dogs with suspected rostral cerebellar ischaemic stroke.
In dogs with ischaemic stroke, a very common site of infarction is the cerebellum. The aim of this study was to characterise neurological signs in relation to infarct topography in dogs with suspected cerebellar ischaemic stroke and to report short-term outcome confined to the hospitalisation period. A retrospective multicentre study of dogs with suspected cerebellar ischaemic stroke examined from 2010-2015 at five veterinary referral hospitals was performed. Findings from clinical, neurological, and paraclinical investigations including magnetic resonance imaging were assessed. Twenty-three dogs, 13 females and 10 males with a median age of 8 years and 8 months, were included in the study. The Cavalier King Charles Spaniel (n = 9) was a commonly represented breed. All ischaemic strokes were located to the vascular territory of the rostral cerebellar artery including four extensive and 19 limited occlusions. The most prominent neurological deficits were gait abnormalities (ataxia with hypermetria n = 11, ataxia without hypermetria n = 4, non-ambulatory n = 6), head tilt (n = 13), nystagmus (n = 8), decreased menace response (n = 7), postural reaction deficits (n = 7), and proprioceptive deficits (n = 5). Neurological signs appeared irrespective of the infarct being classified as extensive or limited. All dogs survived and were discharged within 1-10 days of hospitalisation. Dogs affected by rostral cerebellar ischaemic stroke typically present with a collection of neurological deficits characterised by ataxia, head tilt, and nystagmus irrespective of the specific cerebellar infarct topography. In dogs with peracute to acute onset of these neurological deficits, cerebellar ischaemic stroke should be considered an important differential diagnosis, and neuroimaging investigations are indicated. Although dogs are often severely compromised at presentation, short-term prognosis is excellent and rapid clinical improvement may be observed within the first week following the ischaemic stroke.


CYP24 inhibition preserves 1α,25-dihydroxyvitamin D(3) anti-proliferative signaling in lung cancer cells.
Human lung tumors aberrantly express the 1α,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3))-catabolizing enzyme, CYP24. We hypothesized that CYP24 reduces 1,25(OH)(2)D(3)-mediated transcription and allows lung cancer cells to escape its growth-inhibitory action. To test this, H292 lung cancer cells and the CYP24-selective inhibitor CTA091 were utilized. In H292 cells, CTA091 reduces 1,25(OH)(2)D(3) catabolism, significantly increases 1,25(OH)(2)D(3)-mediated growth inhibition, and increases 1,25(OH)(2)D(3) effects on induced and repressed genes in gene expression profiling studies. Pathway mapping of repressed genes uncovered cell cycle as a predominant 1,25(OH)(2)D(3) target. In H292 cells, 1,25(OH)(2)D(3) significantly decreases cyclin E2 levels and induces G(0)/G(1) arrest. A broader set of cyclins is down-regulated when 1,25(OH)(2)D(3) is combined with CTA091, and cell cycle arrest further increases. Effects of CTA091 on 1,25(OH)(2)D(3) signaling are vitamin D receptor-dependent. These data provide evidence that CYP24 limits 1,25(OH)(2)D(3) anti-proliferative signaling in cancer cells, and suggest that CTA091 may be beneficial in preserving 1,25(OH)(2)D(3) action in lung cancer.


A urinary profile study of dietary phytoestrogens. The identification and mode of metabolism of new isoflavonoids.
The metabolic fate of the dietary isoflavones daidzein and genistein was investigated in human volunteers challenged with soya. Urinary diphenols, isolated by partition chromatography on Sephadex LH-20, were characterized and identified by profile capillary gas chromatography (GC) and electron ionization mass spectrometry (GC-EIMS) analysis of the trimethylsilyl ether (TMS) derivatives. Novel isoflavonic phytoestrogens found in the urine of volunteers were those of tetrahydrodaidzein, dihydrogenistein, 6'-hydroxy-O-demethylangolesin and 2-dehydro-O-demethylangolensin. Other known diphenols identified were those of equal, dehydrodaidzein, O-demethylangolensin, daidzein, genistein, glycitein, and the lignan enterolactone. Two other urinary isomers with a fragmentation pattern closely resembling that of the persilylated TMS ethers of cis/trans-isomers of tetrahydrodaidzein, were characterized based on the elucidation of fragments associated with the loss of a non-phenolic-OTMS functional group in ring-C. These are fragments presented in the persilylated mass spectra of isoflavan-4-ols and isoflav-3-ene-4-ols, demonstrated here by a combination of simple and tandem mass spectrometry study of the deuterated persilylated TMS ethers of dihydrodaidzein. In a similar study we also present the data on the structural identification and fragment elucidation of the keto/enol tautomers of the TMS ether derivatives of the dihydro derivatives of daidzein and genistein, observed in the urine of volunteers and considered probable products of the derivatization process. Finally, the GC and GC-MS data of two unknown isoflavonoids and that of a lignan-like compound are presented together with those of dihydrodaidzein, dihydrogenistein, tetrahydrodaidzein and 2-dehydro-O-demethylangolensin. The latter four were obtained here as products of small scale chemical synthesis in a preliminary study on the tentative identification of urinary isoflavonoids in human volunteers challenged with soya.


Malignant colorectal polyps: endoscopic polypectomy and watchful waiting is not inferior to subsequent bowel resection. A nationwide propensity score-based analysis.
The optimal treatment of patients with malignant colorectal polyps is unsettled. The surgical dilemma following polypectomy is selecting between watchful waiting (WW) and subsequent bowel resection (SBR), but the long-term survival outcomes have not been established yet. This nationwide study compared survival of patients after WW or SBR. Danish nationwide study with 100% follow-up of all patients with malignant colorectal polyps (the Danish Colorectal Cancer Group database) in a 10-year period from 2001 to 2011. All patients' charts and histological reports were individually reviewed. Survival rates were calculated with Cox proportional hazard model after propensity score matching. A total of 692 patients were included (WW, 424 (61.3%), SBR, 268 (38.7%)) with a mean follow-up of 7.5 years (3-188 months). Following propensity score matching, there was no significant difference in overall or disease-free survival (p = 0.344 and p = 0.184) or rate of local recurrence (WW, 7.2%, SBR, 2%, p = 0.052) or distant metastases (WW, 3.3%, SBR, 4.6%, p = 0.77). In the SBR group, there was no residual tumor or lymph node metastases in the resected specimen in 82.5% of the patients. Subsequent bowel resection may not be superior to endoscopic polypectomy and watchful waiting with regard to overall and disease-free survival in patients with malignant colorectal polyps.


Delayed emergence from anesthesia caused by an intraoperative cerebral embolism of a malignant peripheral nerve sheath tumor in a neurofibromatosis type 1 patient: a case report.
Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive soft tissue sarcomas which commonly arise from neurofibromatosis type 1. Lung metastases of the tumors are well-known, but intraoperative cerebral tumor embolisms of MPNSTs have not been reported in literature. A 52-year-old female patient with neurofibromatosis type 1 underwent a right lung partial resection for lung tumors. She was extubated after adequate recovery of spontaneous breathing; however, she could not respond to verbal commands. In the intensive care unit, her neurological examination revealed conjugate eye deviation, right hemiparalysis, and aphasia. Magnetic resonance imaging revealed acute cerebral ischemia, so she underwent an endovascular thrombectomy. The histopathological diagnosis of emboli was a MPNST, which was identical with that of the resected lung tumor. We report the first case of delayed emergence caused by a cerebral tumor embolism of MPNST during partial lung resection.


Prospective study of seasonal patterns in hemostatic factors in older men and their relation to excess winter coronary heart disease deaths.
In England and Wales, approximately 20% extra deaths from coronary heart disease (CHD) occur between December and March, among older people. Circulating concentrations of tissue plasminogen activator (t-PA), von Willebrand factor (VWF) and fibrin D-dimer are associated with arterial disease, and tend to peak in winter. The potential contributions of these hemostatic activation measures to excess winter mortality are unknown. To estimate contributions of hemostatic factors to excess winter mortality. Seasonal patterns in t-PA, VWF and D-dimer were investigated in 4088 men aged 60-79 years from 24 British towns. Data on established coronary risk factors were collected by questionnaire, physical examination and blood sampling. The adjusted mean increase in hemostatic markers during winter months, after adjustment for a range of coronary risk factors, was combined with associations of each marker with CHD mortality obtained from 9 years' follow-up of participants, to predict degree of excess CHD winter mortality. Associations of hemostatic markers with CHD incidence from large meta-analyses were also used. All three markers showed peaks in winter; the adjusted mean increases during winter months were 0.21, 0.15 and 0.12 standard deviations for t-PA, VWF and log(D-dimer), respectively. Predicted excess hazard ratios for winter CHD mortality were 3.0%, 2.4% and 3.1%, respectively, in combination, representing an 8.6% excess. This increased to 14% when applying meta-analysis estimates. Seasonal patterns in three hemostatic markers predict at least 8.6% excess CHD mortality in winter in Great Britain, potentially accounting for over half the excess observed in recent years.


Mitochondrial oxidative phosphorylation: pitfalls and tips in measuring and interpreting enzyme activities.
Mitochondrial oxidative phosphorylation (OXPHOS) is fundamental in all aspects of cellular life in aerobic cells and organisms. It is therefore not surprising that a variety of diseases have been attributed to dysfunction of the OXPHOS enzymes. Assessment of OXPHOS in human samples has proved to be a difficult task over years, even when relying on well-established methods. The complexity and the flexibility of the mitochondrial organization in cells account for a large part in the difficulties encountered in assessing OXPHOS activity. Nevertheless, a careful and detailed analysis of OXPHOS enzyme activity in cells or biopsy samples from patients at risk provides diagnosis of potential OXPHOS deficiency. Problems inherent in the use of human material, mostly the small size of the samples to be analysed, are difficult to resolve. However, cautious handling of these samples permits reasonable confidence to be reached in the interpretation of the data.


Mathematical modelling and simulation of leak detection system in crude oil pipeline.
A first-order differential leak detection model that accurately detects leaks in a crude oil pipeline is presented. This model incorporates a leak factor KL in the axial direction, which is simulated by applying the finite element method of numerical solution using COMSOL multi-physics software. Additionally, the model includes the transport equation for turbulent kinetic energy and the rate of kinetic energy model. Eigenvalues for velocities and pressures were determined and plotted against time for various pipe segments. The system is stable when the Eigenvalue is zero, but a leak is declared when the Eigenvalue for pressure or velocity is less than one. The study shows that pressure measurements are more sensitive parameters for detecting leaks than velocity measurements, and the sinusoidal waveform characterizes leak behaviours for velocity.


A novel spectrophotometric assay for lipase activity utilizing cis-parinaric acid.
A new spectrophotometric assay for determining the activity of acylglycerol hydrolases (lipases, E.C. 184.108.40.206) was developed and optimized for yeast lipase (Candida cylindracea). Studies with porcine pancreatic lipase were also conducted and the influence of various detergents and divalent cations on the assay was evaluated. The assay uses cis-parinaric acid (PnA), a naturally occurring fatty acid that has unique spectroscopic properties, and takes advantage of the reversible binding of fatty acids to bovine serum albumin (BSA). Free PnA has an ultraviolet absorption peak at 321.2 nm. When PnA is bound to BSA, however, the peak shifts to 324.2 nm. The assay mixture contains 6 microM PnA, 1 microM BSA, 75 microM triolein, and 0.3 mM taurocholate in a 50 mM tris-HCl buffer with 1 microM EDTA. The release of oleic acid from triolein is monitored over time by measuring the ratio of optical densities (OD) at 319.0 and 329.0 nm. Initially, there is maximum binding of PnA to BSA, and the OD ratio is approximately 1.0. Upon addition of lipase, PnA is displaced from the BSA by oleic acid released from triolein, and the OD ratio increases to a maximum of about 1.8. However, when calcium is present in the reaction mixture an insoluble calcium-PnA complex forms, resulting in a progressive decrease in OD at both 319.0 and 329.0 nm. The kinetic assay described here is simple, rapid, sensitive, reproducible, inexpensive, and it can be adapted to measure the activity of a variety of calcium-independent lipases. Under similar assay conditions, activities for Candida cylindracea lipase obtained with this assay are similar to those obtained with 14C-labelled triolein.


A case of Acinetobacter calcoaceticus pneumonia.
A case of community acquired pneumonia with Acinetobacter calcoaceticus is presented. Acinetobacter must be considered in the differential diagnosis of Gram negative coccobacillary pneumonia.


Intra-articular botulinum toxin A as an adjunctive therapy for refractory joint pain in patients with rheumatoid arthritis receiving biologics: a report of two cases.
We report the use of intra-articular Botulinum toxin A in two RA patients with persistently painful monoarthritis in ankle/feet joints. Both patients had monarticular pain despite a good response of all other joints to a combination therapy that included anti-tumor necrosis factor therapy. Both patients had failed intra-articular corticosteroid injections and declined surgical options. After a single, "off-label", intra-articular injection of Botulinum toxin A into the right ankle (100 unit) and left first metatarsophalangeal joint (25 unit), respectively, pain and function improved significantly (> or =40%) in both patients. Durable response with improvement of pain and function lasting 15-18 months was noted (follow-up continuing). Ankle joint swelling resolved by 15 months and MTP joint swelling by one month. Intra-articular BoNT/A may be an additional therapeutic option in RA patients with persistent monoarthritis. A randomized study is underway to confirm these findings.


Deaf children with additional disabilities (AD+): CODEPEH recommendations.
Approximately 40% of children with deafness have an additional developmental disorder or major medical problem, which may delay the age of diagnosis of hearing loss and/or require intervention by other professionals. This situation is referred to as "deafness with added disability" (AD+). The reason why the population of hearing-impaired children is more likely to have associated added disabilities is that the risk factors for hearing impairment overlap with those for many other disabilities. These factors can influence various aspects of development, including language acquisition. It is important to check that appropriate care is received, the effectiveness of hearing aids or implants, as well speech therapy intervention strategies, and family adherence to sessions and appointments. The challenge posed by AD+ is early detection, to allow early and appropriate intervention, and the need for fluid transdisciplinary collaboration between all professionals involved, together with the involvement of the family.


Seaweed-based natural ingredients: Stability of phlorotannins during extraction, storage, passage through the gastrointestinal tract and potential incorporation into functional foods.
Adding value to seaweed by extracting their different bioactive compounds and incorporating them into foods represent an interesting and strategic approach to diversify the functional foods offer. However, once harvested, fresh seaweed must overcome a sequence of crucial steps to confer their biological activity. Pre-processing operations and extraction processes, as well as long-term storage, play important roles in improving or decreasing the phlorotannins content. In their way to the gut (biological target), phlorotannins are exposed to the human gastrointestinal tract (GIT), where the physiological pH and digestive enzymes can significantly affect the phlorotannins' stability and thus, alter their biological activity. Besides, the subsequent incorporation into foodstuffs could be limited due to sensory issues, as tannins have been associated with astringency and bitter taste, and thus effective phlorotannins doses may negatively affect the sensory attributes of foods. These drawbacks expose the need of applying smart strategies to develop a final product providing the necessary protective mechanisms to maintain the active molecular form of phlorotannins up to the consumption time, also controlling their release upon arrival to the gut. In this context, the impact of these technological processes (from pre-processing to the passage through the GIT) on phlorotannins stability, as well as the innovative developed approaches to overcome these issues will be deeply discussed in this review. Besides, recent findings related to the phlorotannins' health benefits will be pointed out. Special attention on the potential incorporation of phlorotannins into functional foods will be also put it on.


SAR profiles of spirocyclic nicotinamide derived selective HDAC1/HDAC2 inhibitors (SHI-1:2).
A potent family of spirocyclic nicotinyl aminobenzamide selective HDAC1/HDAC2 inhibitors (SHI-1:2) is profiled. The incorporation of a biaryl zinc-binding motif into a nicotinyl scaffold resulted in enhanced potency and selectivity versus HDAC3, but also imparted hERG activity. It was discovered that increasing polar surface area about the spirocycle attenuates this liability. Compound 12 induced a 4-fold increase in acetylated histone H2B in an HCT-116 xenograft model study with acute exposure, and inhibited tumor growth in a 21-day efficacy study with qd dosing.


Inhibitory effect of opiates on male rat sexual behavior may be mediated by opiate receptors outside the central nervous system.
The importance of opiate receptors outside the central nervous system for the inhibitory actions of morphine on male rat sexual behavior was evaluated. Morphine (10 mg/kg) produced an almost complete inhibition of sexual behavior. This inhibition was antagonized by naloxone at a dose of 1 mg/kg but not at a dose of 0.25 mg/kg. The quaternary opioid antagonist methylnaloxone effectively blocked the inhibitory actions of morphine at a dose of 20 mg/kg but not at a dose of 5 mg/kg. Since the affinity of methylnaloxone for opiate receptors is about 5% of that of naloxone, it may be concluded that both antagonists were about equally effective in inhibiting the effects of morphine. Furthermore, the opiate-like drug loperamide was found to inhibit sexual behavior. This drug acts mainly outside the central nervous system. Its effect was blocked by both naloxone and methylnaloxone, suggesting that opiate receptors are involved. It was also shown that methylnaloxone is unable to block the reinforcing effects of morphine in the conditioned place preference procedure. Because the reinforcing effects of opiates seem to be localized to the central nervous system, it may be proposed that methylnaloxone does not antagonize morphine's central effects. Moreover, loperamide had no effect in the place preference procedure, suggesting that this drug does not act at central opioid receptors. Taken together, these data show that peripheral opioid receptors are responsible for at least some of the inhibitory actions of morphine on male sexual behavior. After treatment with morphine + methylnaloxone, ejaculatory mechanisms were facilitated, reflected in a reduced number of preejaculatory intromissions and a shortened ejaculation latency.(ABSTRACT TRUNCATED AT 250 WORDS)


Novel AT2R agonist, β-Pro7Ang III, is cardio- and vaso-protective in diabetic spontaneously hypertensive rats.
Stimulation of the angiotensin II type 2 receptor (AT2R) evokes protective effects in various cardiovascular diseases. Thus, this study aimed to investigate the effects of AT2R stimulation, with or without AT1R blockade, in a model of hypertension with concomitant type 1 diabetes mellitus (T1DM). Spontaneously hypertensive rats (SHRs) were given either citrate or a single dose of streptozotocin (STZ; 55 mg/kg, i.p.) to induce diabetes. After 4 weeks of diabetes, animals were administered either a vehicle (saline), AT2R agonist, β-Pro7Ang III (0.1 mg/kg/day via osmotic mini-pump), AT1R blocker, candesartan (2 mg/kg/day via drinking water), or a combination of both for a further 8 weeks. β-Pro7Ang III treatment had no effect on blood pressure, but attenuated the significant increase in cardiac interstitial collagen and protein expression of fibrotic and inflammatory markers, and superoxide levels that was evident in diabetic SHRs. These effects were not observed with candesartan, despite its blood pressure lowering effects. Although β-Pro7Ang III had no effect on aortic fibrosis, it significantly attenuated MCP-1 protein expression and superoxide levels when compared to both the non-diabetic and diabetic SHRs, to a similar extent as candesartan. In both the heart and vasculature, the effects of β-Pro7Ang III in combination with candesartan were similar to those of β-Pro7Ang III alone, and superior to candesartan alone. It was concluded that in hypertension with concomitant diabetes, AT2R stimulation with a novel ligand alone, or in combination with AT1R blockade, improved the cardiac and vascular structural changes that were strongly associated with inflammation and oxidative stress, independent of blood pressure regulation.


Design of an actively controlled steerable needle with tendon actuation and FBG-based shape sensing.
This work presents a new steerable needle to facilitate active steering toward predefined target locations. It focuses on mechanical aspects and design choices in relation to the observed response in a tissue phantom. Tip steering with two rotational degrees of freedom was achieved by a tendon actuated ball joint mechanism. During insertion, the flexible cannula bends as a result of asymmetric tip-tissue interaction forces. The stylet was equipped with fiber Bragg gratings to measure the needle shape and tip position during use. A PI-controller was implemented to facilitate steering to predefined targets. During the validation study, nine targets were defined at a depth of 100 mm below the gelatin surface. One was located below the insertion point, the others at a radial offset of 30 mm in each of the eight principle steering directions. Per location, six repetitions were performed. The targeting accuracy was 6.2 ± 1.4 mm (mean ± std). The steering precision was 2.6 ± 1.1 mm. The ability to steer with this new needle steering approach is presented and the mechanical characteristics are discussed for this representative subset of steering directions.


Additives to biological substances. V--The stability of lactose as a carrier for Biological Standards.
The stability of freeze-dried lactose has been studied, by accelerated degradation, after being ampouled under the conditions employed for the preparation of International Standards and Reference Preparations and also under less stringent conditions which might facilitate degradation. The possible formation of the reactive product, 5-hydroxymethyl-furfural, has been monitored over a period of 10 years at temperatures up to 56 degrees C. No evidence has been obtained to suggest that the formation of this compound would present a hazard to the stability of standards prepared by the procedures customarily employed.


Review of animal studies on antimicrobial catheters to prevent catheter-related infections.
Various experimental models have been developed to investigate the pathogenesis of intravascular catheter-related infections. Catheters are implanted either subcutaneously or intravenously, and the test animals are inoculated via either the subcutaneous tissue near the catheter exit site or the bloodstream. Catheters containing antimicrobial agents were investigated in various test models and their efficacy in inhibiting bacterial colonisation and reducing catheter-related infections was demonstrated.


Disease-specific differences in the use of traditional Korean medicine in Korea.
Though traditional Korean medicine plays an important role in the Korean parallel health care system, there is limited information about the preference and usage of traditional Korean medicine compared to Western medicine because they have different disease classification systems. The aim of this study is to determine the relative preference for traditional Korean medicine using data acquired nationwide. Data from the 2008 Korea Health Panel were analyzed to determine the preference of medical services by disease. The use of traditional Korean medicine use is defined by the type of medical institution they used. Disease types, number of visits and out of pocket expenditures were analyzed. Traditional Korean medicine was used in only a small number of cases that were emergencies or hospitalization. However, in terms of outpatient services, traditional Korean medicine was used in 7.8% of all cases and represented 9.9% of total medical costs. Among disease groups, traditional Korean medicine use was higher in patients with nervous system and musculoskeletal system diseases. And patients with musculoskeletal and nervous system diseases such as arthrosis were the most likely to use traditional Korean medicine particularly in an outpatient setting. Korean characteristics of service use resemble the complementary and alternative medicine use in other countries in terms of disease group, and the complementary and alternative medicine should be considered to estimate the burden of disease in countries with parallel health care systems, such as Korea. This is the first study determined the actual preference of traditional Korean medicine for specific chronic diseases.


[Stereotactic body radiation therapy for primary kidney cancer].
The incidence of primary renal cancer is increasing, particularly in elderly patients who may have comorbidities and/or a surgical contraindications. Stereotactic body radiotherapy has primarily been evaluated retrospectively to date. The most commonly used dose schedules are 40Gy in five fractions, 42Gy in three fractions, and 26Gy in one fraction. The results in terms of local control and toxicity are very encouraging. The advantages of stereotactic body radiotherapy compared to thermal ablative treatments are its non-invasive nature, absence of general anesthesia, ability to treat larger lesions, and those close to the renal hilum. Prospective evaluations are still necessary.


Effects of self-administered phencyclidine on regional uptake of 2-deoxy-D-[1-14C]glucose in brain.
Phencyclidine profoundly alters cerebral metabolism in the rat. This study explored whether cerebral metabolic effects of phencyclidine differed when the drug was self-administered by trained rats, compared with when it was given acutely to naive rats. The regional cerebral uptake of 2-deoxy-D-[1-(D14C] glucose (DG) was examined following two injections of phencyclidine (0.5 mg/kg/injection, i.v.) or saline in freely-moving, drug-experienced rats. Naive controls received phencyclidine or saline according to an identical dose regimen. In self-administering and naive rats, phencyclidine produced many of the same effects on uptake of DG, including the following: decreases in the habenula, inferior colliculus, sensory cortical areas and corresponding thalamic relay nuclei; and increases in limbic areas (entorhinal and retrosplenial cortices, subicular areas). Some regions (auditory and motor cortices, medial geniculate body, globus pallidus) showed different effects in self-administering and naive rats. Another study, in which rats were not self-administering phencyclidine, but had histories of treatment with drugs similar to those of the self-administering rats, indicated that chronic exposure to drug accounted for some of the differences. Furthermore, differences between the effects of phencyclidine in self-administering, versus non-self-administering rats with similar histories suggested that activity in some regions of the brain may relate to training in drug self-administration and/or behavior.


Improving Salicornia ramosissima photochemical and biochemical resilience to extreme heatwaves through rhizosphere engineering with Plant Growth-Promoting Bacteria.
The anticipated rise in the length, frequency, and intensity of heatwaves (HW) in the Mediterranean region poses a danger to the crops, as these brief but high-intensity thermal stress events halt plant productivity. This arises the need to develop new eco-friendly sustainable strategies to overcome food demand. Halophytes such as Salicornia ramosissima appear as cash crop candidates, alongside with new biofertilization approaches using Plant Growth Promoting Bacteria (PGPB). In the present work, S. ramosissima plants exposed to heatwave (HW) treatments with and without marine PGPB inoculation is studied to evaluate the physiological responses behind eventual thermal adaptation conditions. Plants exposed to HW inoculated with ACC deaminase and IAA-producing PGPB showed a 50% reduction in the photochemical energy dissipation, when compared to their non-inoculated counterparts, indicating higher light-use efficiency. The observed concomitant increase (76-234%) in several pigments indicates improved inoculated HW-exposed individuals' light harvesting and photoprotection under stressful conditions. This reduction of the physiological stress levels in inoculated plants was also evident by the significant reduction of several antioxidant enzymes as well as of membrane lipid peroxidation products. Additionally, improved membrane stability could also be observed, through the regulation of fatty acid unsaturation levels, decreasing the excessive fluidity imposed by HW treatment. All these improved physiological traits associated with specific PGP traits highlight a key potential of the use of these PGPB consortiums as biofertilizers for S. ramosissima cash crop production in the Mediterranean, where increasing frequency in HW-events is a major drawback to plant production, even to warm-climate plants.


Detection of autoantibodies to survivin and livin in sera from patients with breast cancer.
Survivin and livin are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. Human antibody responses to tumor-associated antigens have been detected, but little is known about the response to survivin and livin in breast cancer patients. We examined the prevalence of anti-survivin and livin antibodies in breast cancer patients with a specific enzyme-linked immunosorbent assay (ELISA) using recombinant protein. Using a cutoff value for positivity determined as the mean absorbance +2SD for healthy control samples, sera from 11 of 46 breast cancer patients (23.9%) were positive by the ELISA using recombinant survivin protein. Of 46 samples from the same breast cancer patients, 15 (32.6%) were positive for anti-livin antibodies. In addition, 24 (52.2%) were positive for 1 or both ELISAs using the respective proteins. Intensity of anti-livin antibody responses did not correlate with intensity of anti-survivin responses. Anti-livin antibodies were detected in sera from breast cancer patients by an anti-livin ELISA using full-length recombinant livin protein. Like survivin, livin may act as a major cancer antigen in breast cancer patients.


Development and validation of the Slovenian drug-related problem classification system based on the PCNE classification V 6.2.
Background Classifying drug-related problems increases pharmacists' awareness of patients' drug-related needs and supports the development of counselling skills through increased awareness of the nature and frequency of drug-related problems. No standardised classification system was used in daily pharmacy practice in Slovenia. Objective To translate, upgrade and validate the Pharmaceutical Care Network Europe (PCNE) classification V 6.2 for use in Slovenian community pharmacies. Setting Expert panel meetings at the Faculty of Pharmacy and home-based classification validation. Methods The PCNE classification V 6.2 was translated to Slovenian language by forward-backward translation procedure. An expert panel consisting of nine practicing pharmacists upgraded the content of the translated version. Thirty-one community pharmacists validated this version with the PCNE set of 18 patient cases by coding problems, risk factors and interventions they believed were present in each case. The expert panel discussed the results and upgraded the classification accordingly. Afterwards, 33 community pharmacists validated the upgraded version with a set of 40 actual Slovenian pharmacy patient cases. Based on the results, the expert panel formed a final version of the classification. Main outcome measure Coding consistency between community pharmacists. Results The expert panel performed some major modifications to the PCNE classification V 6.2: the potential problem was added as a sub domain to problems domain; the term adverse drug event was used instead of adverse drug reaction; the causes domain was renamed to risk factors and its sub domains were organized into prescribing, dispensing and use of drugs; dispensing errors were specified; use of drugs was organized into intentional and unintentional use of drugs; the sub domains in the interventions domain were divided according to the communication and agreement with the prescriber. The average coding consistencies in the first validation study were 75 % (±16 %) for problems, 74 % (±20 %) for risk factors and 94 % (±10 %) for interventions. The average coding consistencies in the second validation study were 83 % (±16 %) for problems, 85 % (±17 %) for risk factors and 80 % (±20 %) for interventions. Conclusion A drug-related problem classification for use in Slovenian community pharmacies was developed based on The PCNE classification V 6.2. The validation studies demonstrated high coding consistencies.


Public-private partnership in the Portuguese health sector.
Since 2001, in Portugal, constant reforms in hospital management have accompanied the transformations in the management models applied to public administration, intending to ensure a higher quality of services and, simultaneously, a more significant economic efficiency. This study aims to analyse, for the period between 2012 and 2021, the economic and financial results (value-for-money) of the PPP model, compared with the public management hospitals (PMH). It used a mixed research approach based on multiple case studies and archival research. As the main results, it was found that: i) the PPP model, applied to the health sector, appears to be advantageous, not only regarding the economic and financial results but also concerning the quality of service provision; and ii) despite the value-for-money generated by the PPP model, the lower operating costs and the superior performance in comparison with PMH, the government has permanently opted to revert from a PPP model to a PMH model. This study concluded that the hospital management model is instead seen as an instrumentalised political instrument than a management tool that could generate savings for the taxpayers. Several practical implications are presented.


Bone biomarkers in de novo renal transplant recipients.
Successful kidney transplantation (partly) corrects the physiologic and metabolic abnormalities driving chronic kidney disease - mineral and bone disorders. At the same time, renal transplant recipients are exposed to immunosuppressive agents that may affect bone metabolism. Bone biomarkers have been suggested as surrogates of or adjuncts to bone biopsy and imaging techniques to assess bone health and to classify risk of bone loss and fractures. Bone biomarkers may be classified as circulating factors that affect bone metabolism (commonly referred to as bone metabolism markers) or that reflect bone cell number and/or activity (commonly referred to as bone turnover markers). A growing body of evidence shows that successful renal transplantation has a major impact on both bone metabolism and bone turnover. Analytical issues, including the cross-reactivity with fragments, complicate the interpretation of bone biomarkers, especially in the setting of a rapid changing kidney function, as is the case after successful renal transplantation. Overall, bone turnover seems to decline following renal transplantation, but inter-individual variability is substantial. Preliminary evidence indicates that bone biomarkers may be useful in guiding mineral and bone therapy in renal transplant recipients.


Neuroendocrine tumors of the pancreas: a retrospective single-center analysis using the ENETS TNM-classification and immunohistochemical markers for risk stratification.
This study was performed to assess the 2006 introduced ENETS TNM-classification with respect to patient survival and surgical approach for patients who underwent surgery for a neuroendocrine tumor of the pancreas (PNET). Between 2001 and 2010 38 patients after resection of a PNET were investigated regarding tumor localization and size. Further, patient survival with regards to the new TNM-classification, the operation methods and immunohistochemical markers was analyzed. The estimated mean survival time of the 38 patients was 91 ± 10 months (female 116 ± 9, male 56 ± 14 months; p = 0.008). The 5-year survival rate was 63.9%. Patient survival differed significantly depending on tumor size (pT1 107 ± 13, pT2 94 ± 16, pT3 44 ± 7 and pT4 18 ± 14 months; P = 0.006). Patients without lymph node metastasis survived significantly longer compared to patients with positive lymph node status (108 ± 9 vs. 19 ± 5 months; P < 0.001). However, survival in patients with and without distant metastasis did not differ significantly (92 ± 11 vs. 80 ± 23 months; P = 0.876). Further, the tumor grading significantly influenced patient survival (G1 111 ± 12, G2 68 ± 12 and G3 21 ± 14 months; P = 0.037). As part of the TNM-classification especially lymph node status and also tumor size and grading were identified as important factors determining patient survival. Further, gender was demonstrated to significantly influence survival time. If an R0 resection was achieved in patients with distant metastases patient survival was comparable to patients without metastasis.


Synthesis and molecular recognition of carbohydrate-centered multivalent glycoclusters by a plant lectin RCA120.
Water soluble and lectin-recognizable carbohydrate-centered glycoclusters were prepared efficiently by the Huisgen 1,3-cycloaddition reaction of methyl-2,3,4,6-tetra-O-propargyl beta-D-galactopyranoside with 2-azidoethyl glycosides of lactose and N-acetyllactosamine. Their binding by a plant lectin RCA120 was examined by capillary affinity electrophoresis using fluorescence-labeled asialoglycans from human alpha1-acid glycoprotein. The glycoclusters showed 400-fold stronger inhibitory effect than free lactose, manifesting strong multivalency effect.


A highly basic histone H4 domain bound to the sharply bent region of nucleosomal DNA.
A nucleosomal core particle is composed of two each of histones H2A, H2B, H3 and H4 located inside the particle with approximately 47 base pairs (bp) of DNA wrapped around the octamer in about 1.8 turns of a left-handed superhelix. The path of the superhelix is not smooth; the DNA is sharply bent, or kinked, at positions symmetrically disposed at a distance of about one and four double-helical turns in both directions from the nucleosomal dyad axis (designated as sites +/- 1 and +/- 4 respectively). This non-uniform bending is considered archetypal to other DNA-protein complexes, but its mechanism is not clear (reviewed in ref. 4). DNA-histone chemical cross-linking within the core particle has revealed strong binding of each of the two histone H4 molecules to DNA at a distance of 1.5 helical turns either side of the nucleosomal dyad axis (sites +/- 1.5). In each of these sites, a single flexible domain of H4 was previously shown to contact three points, at about nucleotides 55 and 65 on one strand and nucleotide 88 on the complementary strand, numbering from the 5' terminus of each 147-base strand; these three locations are closely juxtaposed across the highly compressed minor and major grooves (Fig. 1). Here we report that the amino-acid residue of histone H4 cross-linked at the 1.5 site is histidine-18, embedded in a highly basic cluster Lys-Arg-His-Arg-Lys-Val-Leu-Arg which is probably involved in the sharp bending of the DNA double helix at the +/- 1 sites.


Drug-Drug Interactions With Novel All Oral Interferon-Free Antiviral Agents in a Large Real-World Cohort.
With the approval of direct-acting antivirals (DAAs), the management of drug-drug interactions (DDIs) has become an important challenge while treating individuals with hepatitis C. To date, the potential of causing DDIs for the recently approved DAAs has not been systematically investigated. We aimed to assess the clinical significance of DDI between the regular outpatient medications and DAA therapies in a large real-world cohort. Overall, 261 hepatitis C virus monoinfected patients who were selected for DAA therapy at 2 intervals between 2011 and 2014 were asked about their regular outpatient medications. The potential for DDIs between all these drugs and sofosbuvir/ribavirin, ledipasvir/sofosbuvir, sofosbuvir/daclatasvir, sofosbuvir/simeprevir, ombitasvir/paritaprevir/ritonavir ± dasabuvir as well as boceprevir and telaprevir triple therapy was assessed using www.hep-druginteractions.org and the relevant prescribing information. The 261 patients took a median number of 2 drugs (range 0-15); 20% of patients did not take any medication. Sofosbuvir/ribavirin had the lowest risk to cause a potentially significant DDI (9.6%). In contrast, for ombitasvir/paritaprevir/ritonavir ± dasabuvir potentially significant DDIs could be expected in 66.3% of the patients. Significant DDIs for sofosbuvir/simeprevir would be expected in 31.4%, for sofosbuvir/daclatasvir in 36.8%, and for sofosbuvir/ledipasvir in 40.2%. Proton pump inhibitors, thyroid hormones, and dihydropyridine derivatives were frequently used and presented a risk of interacting with the antiviral regimen. A significant number of patients are at risk for DDIs if treated with the recently approved DAA regimens. A careful evaluation of potential DDI is essential to prevent adverse effects or unnecessary risk of treatment failure.


Finding the productivity standard in your acuity system.
What is a realistic patient workload for nurses to handle on a given shift? Should the workload vary from shift-to-shift and nurse-to-nurse? This article describes how (1) the authors identified the standard inherent in their patient classification system; (2) the system is used to establish department workload standards; (3) managers comply with the established standard; and (4) nursing costs are affected when compliance with the standard is not achieved.


MicroRNAs and fracture healing: Pre-clinical studies.
During the past several years, pre-clinical experiments have established that microRNAs (miRNAs), small non-coding RNAs, serve as key regulatory molecules of fracture healing. Their easy modulation with agonists and antagonists make them highly desirable targets for future therapeutic strategies, especially for pathophysiologic fractures that either do not heal (nonunions) or are delayed. It is now well documented that these problematic fractures lead to human suffering and impairment of life quality. Additionally, financial difficulties are also encountered as work productivity decreases and income is reduced. Moreover, targeting miRNAs may also be an avenue to enhancing normal physiological fracture healing. Herein we present the most current knowledge of the involvement of miRNAs during fracture healing in pre-clinical studies. Following a brief description on the nature of miRNAs and of the fracture healing process, we present data from studies focusing specifically, on miRNA regulation of osteoblast differentiation and osteogenesis (within the context of known signaling pathways), chondrocytes, angiogenesis, and apoptosis, all critical to successful bone repair. Further, we also discuss miRNAs and exosomes. We hope that this manuscript serves as a comprehensive review that will facilitate basic/translational scientists in the orthopaedic arena to realize and further decipher the biological and future therapeutic impact of these small regulatory RNA molecules, especially as they relate to the molecular events of each of the major phases of fracture healing.


Vasopressin and norepinephrine stimulation of inositol phosphate accumulation in rat hepatocytes are modified differently by protein f1nase C and protein kinase A.
Rat hepatocytes were maintained in primary monolayer culture for 24 h in the presence of serum. Treatment of hepatocytes with 1 microM 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) for 5-15 min increased membrane-associated protein kinase C activity and concomitantly decreased soluble activity. Membrane protein kinase C activity returned to basal values within 1 h then decreased by more than 50% within 2 h. Prolonged (2-18 h) incubation with PMA did not further decrease protein kinase C activity. Pretreatment of hepatocytes with PMA for 5-15 min had little effect on the subsequent actions of 100 nM vasopressin but abolished the stimulation of inositol phosphate accumulation by 3 nM vasopressin and 20 microM norepinephrine. Long-term exposure (2-18 h) of hepatocytes to 1 microM PMA actually enhanced the effects of vasopressin and 20 microM norepinephrine. The stimulation by norepinephrine (20 microM) of inositol phosphate accumulation was abolished by the alpha 1-adrenergic antagonist prazosin (1 microM), whereas the beta-adrenergic antagonist propranolol (30 microM) had little effect. Addition of 8Br-cAMP (100 microM) or glucagon (10 nM) for 5 min or 8 h had no significant effect alone, but enhanced the subsequent vasopressin stimulation of inositol phosphate accumulation. There was no effect of 8Br-cAMP or glucagon on norepinephrine stimulation of phosphoinositide breakdown. These data indicate that the stimulation of phospholipase C activity in rat hepatocytes by 3 nM vasopressin is enhanced by cyclic AMP-dependent kinase but inhibited by protein kinase C. In contrast, down regulation of protein kinase C markedly enhanced the maximal phosphoinositide response due to both vasopressin and norepinephrine.


Stepwise dehydration of Cd-exchanged levyne: thermal stability and structural modifications.
Zeolites show remarkable properties that can be tuned through cation exchange of their original extraframework content. In this respect, the response of the modified zeolite to the heating stimuli, in terms of structural modifications and thermal stability, can drastically change and is, therefore, an important factor to consider. In this study, the dehydration mechanism of a natural levyne previously exchanged with Cd2+ has been monitored in situ by single crystal X-ray diffraction. The initial dehydration trend between 50 and 175 °C is similar to that observed for the pristine material, levyne-Ca. The water loss is accompanied by extraframework cation migration within the zeolitic cavities and the unit-cell volume slightly contracts from 3503.8(1) to 3467.8(6) Å3. From 200 to 250 °C, a pronounced drop of the unit-cell volume (- 7%) is observed. The dehydrated structure at 250 °C corresponds to levyne B topology of natural levyne, characterized by the statistical rupture of the T-O-T bonds of the double six-ring membered cage. However, in contrast to levyne-Ca, the fraction of broken connections reached 50% instead of 37%, and no additional structural modifications were detected up to 350 °C. At 400 °C, diffraction data pointed to the onset of the structural collapse. At this temperature, the measured unit-cell volume was 8% smaller compared to that of the RT structure. The corresponding contracted structure did not rehydrate after exposure to humid conditions for 21 days. The online version contains supplementary material available at 10.1007/s00269-021-01146-6.


Lipid Nanoparticles Optimized for Targeting and Release of Nucleic Acid.
Lipid nanoparticles (LNPs) are currently the most promising clinical nucleic acids drug delivery vehicles. LNPs prevent the degradation of cargo nucleic acids during blood circulation. Upon entry into the cell, specific components of the lipid nanoparticles can promote the endosomal escape of nucleic acids. These are the basic properties of lipid nanoparticles as nucleic acid carriers. As LNPs exhibit hepatic aggregation characteristics, enhancing targeting out of the liver is a crucial way to improve LNPs administrated in vivo. Meanwhile, endosomal escape of nucleic acids loaded in LNPs is often considered inadequate, and therefore, much effort is devoted to enhancing the intracellular release efficiency of nucleic acids. Here, different strategies to efficiently deliver nucleic acid delivery from LNPs are concluded and their mechanisms are investigated. In addition, based on the information on LNPs that are in clinical trials or have completed clinical trials, the issues that are necessary to be approached in the clinical translation of LNPs are discussed, which it is hoped will shed light on the development of LNP nucleic acid drugs.


Influences of morphology and topography of motoneurons and muscle spindle afferents on amplitude of single fiber excitatory postsynaptic potentials in cat.
Excitatory postsynaptic potentials (e.p.s. p.s.) elicited by impulses in single muscle spindle afferent fibers from the medial gastrocnemius (m.g.) muscle were recorded intracellularly from homonymous and heteronymous motoneurons in order to study factors that influence the amplitudes of such responses. Impulses in large afferent fibers elicited larger single-fiber e.p.s.p.s. than those in smaller afferents. Mean e.p.s.p. amplitudes were related exponentially to afferent conduction velocities of both Ia and spindle group II fibers. The closer a motoneuron was to the spinal entry point of an afferent fiber, the larger was the mean e.p.s.p. amplitude evoked in it. Impulses in the same afferent fiber elicited larger e.p.s.p.s in small than large motoneurons when the two cells were located at the same craniocaudal levels. Other factors being equal, the single-fiber e.p.s.p.s evoked in homonymous and heteronymous motoneurons were approximately equal in amplitude. Relatively simple morphological and topographical explanations for the findings were advanced and their functional significance for orderly recruitment in partitioned and unpartitioned reflexes was described.


Unusual group migration and C(sp3)-H activation leading to stable metallacycles in the reactions of Cp*IrS2C2B10H10 and aryl azides.
The thermal or photochemical reactions of Cp*IrS(2)C(2)B(10)H(10) (1) and aryl azides lead to C-C coupling via C(sp(3))-H activation in 2 as well as the formation of C-S bonds and new-type SSN pincer ligands in 3-8 through ortho-substituted electron-withdrawing group migration over an aryl ring.


Alpha 1-microglobulin in urine as an indicator of renal tubular damage caused by environmental cadmium exposure.
Alpha1-Microglobulin (alpha 1-m) was determined in the urine of both the cadmium-exposed and nonexposed subjects and was compared with those of other urinary parameters such as beta 2-microglobulin (beta 2-m), glucose, total protein or amino acids. Large amounts of alpha 1-m were detected in the urine of cadmium-exposed subjects. There were significant correlations between alpha 1-m and other urinary parameters. The sensitivity and specificity of both alpha 1-m and beta 2-m as indices of tubular dysfunction caused by cadmium was similar. It was concluded that alpha 1-m in urine seemed to be a useful indicator of renal tubular dysfunction caused by cadmium.


From measuring to removing to recovering phosphorus in water management in the Netherlands: Challenges for science-based innovation.
The research question of this paper is: Which lessons for enabling a full scale recovery of phosphorus from waste water can be drawn from 1. an analysis of paradigmatic changes in scientific knowledge on P? In what way can these lessons inform scientists and (innovation) policy makers to stimulate a transition towards full scale P recovery from waste water? The paper draws upon a longitudinal case study of water management in the Netherlands and how it has dealt in science and policy with P, since approximately 1960 until now. It discusses and analyzes paradigm shifts in science and changes in the underlying organization of R&D on P. It ends with additional lessons on the most optimal organization of research and innovation infrastructures for the full scale recovery of P. 2. an analysis of the changes in the organization of R&D for phosphorus (P). Challenges relate to 1. the lack of a market for recovered phosphate in Europe, 2. the lack of experience of water managers in commercializing their findings and products 3. the lack of democratic legitimacy of regional water authorities 4. limitations put on the activities of water managers due to legislation and regulatory frameworks.


Studying mucin secretion from human bronchial epithelial cell primary cultures.
Mucin secretion is regulated by extracellular signaling molecules emanating from local, neuronal, or endocrine sources. Quantifying the rate of this secretion is important to understanding how the exocytic process is regulated, and also how goblet/mucous cells synthesize and release mucins under control and pathological conditions. Consequently, measuring mucins in a quantitatively accurate manner is the key to many experiments addressing these issues. This paper describes procedures used to determine agonist-induced mucin secretion from goblet cells in human bronchial epithelial (HBE) cell cultures. It begins with primary epithelial cell culture, offers methods for purifying MUC5AC and MUC5B mucins for standards, and describes five different microtiter plate binding assays which use various probes for mucins. A polymeric mucin-specific antibody is used in standard and sandwich ELISA formats for two assays while the others target the extensive glycosylated domains of mucins with lectin, periodate oxidation, and antibody-based probes. Comparing the data derived from the different assays applied to the same set of samples of HBE cell cultures indicates a qualitative agreement between baseline and agonist stimulated mucin release; however, the polymeric mucin-specific assays yield substantially lower values than the assays using non-specific molecular reporters. These results indicate that the more nonspecific assays are suitable to assess overall secretory responses by goblet cells, but are likely unsuited for specific measurements of polymeric mucins, per se.