(S)-homoquisqualate: a potent agonist at the glutamate metabotropic receptor.
The synthetic quisqualate analogue, (S)-homoquisqualate was examined for activity at the glutamate metabotropic receptor, in relation to its ability to stimulate phosphoinositide hydrolysis in rat pup cerebro-cortical slices. The compound produced a robust increase in hydrolysis (EC50 = 50.2 +/- 1.6 microM), which, in common with responses to quisqualate and (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate ((1S,3R)-ACPD), was antagonized uncompetitively by L-2-amino-3-phosphonopropionate (L-AP3). In contrast to quisqualate which exhibits low efficacy, (S)-homoquisqualate behaves as a full agonist at the metabotropic receptor.


Evaluation of standard breast ultrasonography by adding two-dimensional and three-dimensional shear wave elastography: a prospective, multicenter trial.
To reduce the number of biopsies performed on benign breast lesions categorized as BI-RADS 4-5, we investigated the diagnostic performance of combined two-dimensional and three-dimensional shear wave elastography (2D + 3D SWE) with standard breast ultrasonography (US) for the BI-RADS assessment of breast lesions. A total of 897 breast lesions, categorized as BI-RADS 3-5, were subjected to standard breast US and supplemented by 2D SWE only and 2D + 3D SWE analysis. Based on the malignancy rate of less than 2% for BI-RADS 3, lesions assessed by standard breast US were reclassified with SWE assessment. After standard breast US evaluation, 268 (46.1%) participants underwent benign biopsies in BI-RADS 4-5 lesions. By using separated cutoffs for upstaging BI-RADS 3 at 120 kPa and downstaging BI-RADS 4a at 90 kPa in 2D + 3D SWE reclassification, 123 (21.2%) participants underwent benign biopsy, resulting in a 54.1% reduction (123 versus 268). Combining 2D + 3D SWE with standard breast US for reclassification of BI-RADS lesions may achieve a reduction in benign biopsies in BI-RADS 4-5 lesions without sacrificing sensitivity unacceptably. Combining 2D + 3D SWE with US effectively reduces benign biopsies in breast lesions with categories 4-5, potentially improving diagnostic accuracy of BI-RADS assessment for patients with breast lesions. ChiCTR1900026556 KEY POINTS: • Reduce benign biopsy is necessary in breast lesions with BI-RADS 4-5 category. • A reduction of 54.1% on benign biopsies in BI-RADS 4-5 lesions was achieved using 2D + 3D SWE reclassification. • Adding 2D + 3D SWE to standard breast US improved the diagnostic performance of BI-RADS assessment on breast lesions: specificity increased from 54 to 79%, and PPV increased from 54 to 71%, with slight loss in sensitivity (97.2% versus 98.7%) and NPV (98.1% versus 98.7%).


The Mantle Transcriptome of Chamelea gallina (Mollusca: Bivalvia) and Shell Biomineralization.
The striped venus Chamelea gallina is a bivalve mollusc that represents one of the most important fishery resources of the Adriatic Sea. In this work, we investigated for the first time the ability of this species to modulate the expression of genes encoding proteins involved in biomineralization process in response to biotic and abiotic factors. We provided the first comprehensive transcriptome from the mantle tissue of clams collected in two sampling sites located along the Italian Adriatic coast and characterized by different environmental features. Moreover, the assessment of environmental parameters, scanning electron microscopy (SEM), and X-ray diffraction (XRD) measurements on valves were conducted to better contextualize RNA sequencing (RNA-Seq) data. Functional annotation of differentially expressed genes (DEGs) and SEM observations highlighted a different shell mineralization behaviour in C. gallina clams collected from two selected sites characterized by diverse environmental parameters.


The observational study of delayed wound healing after tooth extraction in patients receiving oral bisphosphonate therapy.
In this study, we investigated whether such a discontinuation of oral bisphosphonate (BP) for 3 months might influence the incidence of BP-related osteonecrosis of the jaw (BRONJ) and wound healing after tooth extraction in patients receiving oral BP therapy. There were a total of 434 teeth in 201 patients (18 males and 183 females). The patients were divided into two groups depending on whether or not they underwent a 3-month discontinuation of BP therapy (BP- and BP+) before tooth extraction. In this observational study investigated delayed wound healing after tooth extraction in patients receiving oral BP therapy. In all cases of the BP- group, there were no BRONJ although there was delayed wound healing in two cases. However, in one case of the BP+ group, oral BP was continued because it was deemed high risk to discontinue treatment by the patient's physician. In this case, an intraoral fistula was still present with bone exposure at 120 weeks after extraction (BRONJ stage 1). This study supports the idea of a drug holiday and encourages further clinical research on this topic of tooth extraction in patients receiving oral BP therapy.


Dose to specific subregions of pelvic bone marrow defined with FDG-PET as a predictor of hematologic nadirs during concomitant chemoradiation in anal cancer patients.
To test the hypothesis that irradiated volume of specific subregions of pelvic active bone marrow as detected by (18)FDG-PET may be a predictor of decreased blood cells nadirs in anal cancer patients undergoing concurrent chemoradiation, we analyzed 44 patients submitted to IMRT and concurrent chemotherapy. Several bony structures were defined: pelvic and lumbar-sacral (LSBM), lower pelvis (LPBM) and iliac (IBM) bone marrow. Active BM was characterized employing (18)FDG-PET and characterized in all subregions as the volume having standard uptake values (SUVs) higher than SUVmean. All other regions were defined as inactive BM. On dose-volume histograms, dosimetric parameters were taken. Endpoints included white blood cell count (WBC), absolute neutrophil count (ANC), hemoglobin (Hb) and platelet (Plt) nadirs. Generalized linear modeling was used to find correlations between dosimetric variables and blood cells nadirs. WBC nadir was significantly correlated with LSBM mean dose (β = -1.852; 95 % CI -3.205/-0.500; p = 0.009), V10 (β = -2.153; 95 % CI -4.263/-0.721; p = 0.002), V20 (β = -2.081; 95 % CI -4.880/-0.112; p = 0.003), V30 (β = -1.971; 95 % CI -4.748/-0.090; p = 0.023) and IBM V10 (β = -0.073; 95 % CI -0.106/-0.023; p = 0.016). ANC nadir found to be significantly associated with LSBM V10 (β = -1.878; 95 % CI -4.799/-0.643; p = 0.025), V20 (β = -1.765; 95 % CI -4.050/-0.613; p = 0.030) and IBM V10 (β = -0.039; 95 % CI -0.066/-0.010; p = 0.027). Borderline significance was found for correlation between Plt nadir and LSBM V30 (β = -0.056; 95 % CI -2.748/-0.187; p = 0.060), V40 (β = -0.059; 95 % CI -3.112/-0.150; p = 0.060) and IBM V30 (β = -0.028; 95 % CI -0.074/-0.023; p = 0.056). No inactive BM subsites were found to be correlated with any blood cell nadir. (18)FDG-PET is able to define active bone marrow within pelvic osseous structures. LSBM is the strongest predictor of decreased blood cells nadirs in anal cancer patients undergoing concurrent chemoradiation.


Quantitative measurements on wetted thin layer chromatography plates using a charge coupled device camera.
This paper presents the first study of imaging of spots on thin-layer chromatographic plates whilst still wet with solvent. Imaging and quantification of Sudan II after development with dichloromethane was carried out in both reflectance and transmission modes, using a charge coupled device (CCD) camera. The relationship between peak area and sample loading was established at low sample loading, and found to be linear over an order of magnitude for both wet and dry modes with r2-values > 0.99. All data processing was carried out using the Beer-Lambert equation. Curvature at high loadings in the plots of integrated absorbance as a function of sample loading was accounted for using an empirical expression designed for use with the Kubelka-Munk treatment and apparent absorbance of the stationary phase due to scattering. Results are consistent with an effective pathlength significantly longer than the thickness of the sorbent layer. The limit of detection on a dry plate (0.5 ng) was found to be lower than on a wetted plate (2 ng). Precision was found to be 1-4% RSD intra-plate and 8-14% RSD inter-plate. Results are compared with quantification of the same analyte on dried plates.


Effects of footshocks on anxiety-like behavior and mRNA levels of precursor peptides for corticotropin releasing factor and opioids in the forebrain of the rat.
Corticotropin releasing factor (CRF) and dynorphin are neuropeptides that are associated with the negative emotional states. Experimental evidence indicates that dynorphin neurons located in the nucleus accumbens and CRF neurons in the bed nucleus of the stria terminalis (BST) and the central nucleus of the amygdala (CeA) mediate anxiety-like behaviors immediately after the stressful experience (24-48h). The present study was done to evaluate if changes in the levels of the mRNA for these peptides in the striatum, BST, and CeA were associated with the long-lasting avoidance of novelty, a measure of an anxiety-like state, in a subset of rats exposed to unpredictable and moderately intense footshocks (5×2s of 1.5mA). Shocked rats with enhanced fear to a novel tone 24h after the footshocks (high responders; HR) displayed long-lasting avoidance in the elevated T-maze whereas shocked rats with low levels of acute fear (low responders; LR) had low levels of avoidance similar to nonshocked rats. An increase in the level of proCRF mRNA was detected in the CeA of the HR compared to LR and nonshocked rats but not in other areas of the brain sampled. In contrast, prodynorphin and proenkephalin mRNA levels in the striatum, BST and CeA were not different between HR, LR and nonshocked rats. This study provides evidence that CRF neurons in the CeA may play a role in the anxiety-like state produced in a subset of rats exposed to footshocks.


A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia.
A method to measure cutaneous hyperalgesia to thermal stimulation in unrestrained animals is described. The testing paradigm uses an automated detection of the behavioral end-point; repeated testing does not contribute to the development of the observed hyperalgesia. Carrageenan-induced inflammation resulted in significantly shorter paw withdrawal latencies as compared to saline-treated paws and these latency changes corresponded to a decreased thermal nociceptive threshold. Both the thermal method and the Randall-Selitto mechanical method detected dose-related hyperalgesia and its blockade by either morphine or indomethacin. However, the thermal method showed greater bioassay sensitivity and allowed for the measurement of other behavioral parameters in addition to the nociceptive threshold.


Awareness of the need for safe storage of methadone at home is not improved by the use of protocols on recording information giving.
Methadone is a synthetic, narcotic analgesic used in the treatment of drug misuse. Tragedies involving children being poisoned by the accidental ingestion of methadone are no longer a rare occurrence. Following an audit of the effectiveness of the provision and recall of information to patients attending an NHS Methadone Clinic a protocol was introduced to ensure that staff documented the provision of such information and patients gave a written confirmation that they had received the information. The study was undertaken in the setting of an NHS methadone clinic with the aim of re- auditing the storage of methadone at home following the introduction of the new protocols. 174 patients completed an anonymous questionnaire regarding where they store methadone at home and whether they recall being given advice about safe storage. Community pharmacists were contacted by telephone to assess the level of advice given to methadone patients regarding safety. Only 49 (28.2%) patients recalled being given advice about safe storage, 24 (13.8%) recalled that information was provided by clinic staff. 170 (97.7%) patients regard methadone as being dangerous. (28.2%). Methadone is most commonly stored in a cupboard (37.9%). All methadone is dispensed in a bottle with a child resistant cap on it. All patients reported they stored their methadone in the original bottle provided by the pharmacist. Recall of information on safety issues is very poor. Provision of written as well as verbal information is needed. The use of printed safety information cards which patients can take away for future reference may be of use. It is the responsibility of health professionals to ensure they provide information and advice to methadone users on the safe storage of their methadone at home.


The long non-coding RNA, GAS5, enhances gefitinib-induced cell death in innate EGFR tyrosine kinase inhibitor-resistant lung adenocarcinoma cells with wide-type EGFR via downregulation of the IGF-1R expression.
Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) are approved for patients with recurrent non-small cell lung cancer (NSCLC). However, the efficacy of EGFR-TKIs in NSCLC therapy is limited by primary and acquired resistance. Recent studies have revealed that long non-coding RNAs (LncRNA) may be involved in EGFR-TKI resistance. Therefore, a better understanding of the interactive mechanisms underlying LncRNA-mediated EGFR-TKIs resistance may help us to improve clinical response rates. To investigate the expression of growth arrest-specific 5 (GAS5) in lung adenocarcinoma, we performed real-time reverse-transcriptase polymerase chain reaction. The correlation between GAS5 expression levels and the samples' clinicopathological features was also analyzed. Primary resistance to EGFR-TKIs was identified in the human lung adenocarcinoma cell line A549. Plasmid vectors were used to overexpress GAS5 in A549 cells. MTT (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide) colony formation assays and EdU (5-ethynyl-2'-deoxyuridine) assays were used to assess cell proliferation, and flow-cytometric analysis was used to evaluate the apoptosis rate. The expression levels of our target proteins, namely, EGFR, p-EGFR, ERK, p-ERK, Akt, p-Akt, IGF-1R (insulin-like growth factor 1 receptor), and p-IGF-1R, were analyzed by western blotting. A549 cells transfected with pcDNA-GAS5 were injected into nude mice. The transplanted mice were treated with gefitinib to study the effect of GAS5 on the resistance to EGFR-TKIs in vivo. Our results showed that GAS5 was significantly downregulated in lung adenocarcinoma tissues compared with the paired adjacent non-tumorous tissue samples. Furthermore, lower GAS5 expression levels were associated with larger tumor sizes, poor tumor differentiation, and advanced pathological stages. However, GAS5 was almost equally expressed between benign tumors compared with the adjacent normal tissues. GAS5 was also overexpressed in EGFR-TKI sensitive cell lines compared with the resistant cell line. Using MTT, EdU incorporation, and colony formation assays, we showed that GAS5-expressing A549 cells displayed an elevated level of cell death. In addition to its pro-apoptotic effect in the A549 cell line, GAS5 overexpression also suppressed the growth of A549-derived tumors in nude mice treated with gefitinib. GAS5 overexpression was inversely correlated with the expression of the EGFR pathway and IGF-1R proteins. Collectively, our results indicated that GAS5 LncRNA may represent a potential biomarker for the diagnosis of lung adenocarcinoma and that GAS5 might play a novel role in the development of the resistance to gefitinib, which could be reversed by overexpressing GAS5.


Survivorship care planning after participation in communication skills training intervention for a consultation about lymphoma survivorship.
A survivorship care plan refers to a written summary of the treatment received and recommendations regarding surveillance and management of late effects. To provide evaluation of a communication skills training (CST) intervention to enhance the transition of lymphoma survivors to cancer survivorship. Nineteen oncologists specializing in lymphoma treatment were recruited and completed a survivorship CST workshop, and two standardized patient assessments (SPAs), one pretraining and one posttraining. Significant improvements in SPA scores were observed in six of the seven SPA assessment categories: use of survivorship care plan, review of disease and treatment details, long-term effects, potential late effects, specific physician recommendations, and additional health maintenance recommendations. The intervention had significant effects on physicians' uptake of new strategies and skills, as measured through pre- and posttraining SPAs, as well as on the physicians' self-efficacy about having these conversations.


An assay to measure antigen-specific immune complexes in food-allergy patients.
An allergen- and isotype-specific assay to quantify the presence of serum immune complexes is described. This assay is based on a two-site recognition system so that only molecular complexes containing both antigen and antibody are counted. In this study affinity-purified antibodies to egg and milk whey were used on paper discs in conjunction with a soluble, labeled anti-immunoglobulin (IgE or IgG specific). To evaluate the assay, immune complexes were prepared in vitro by mixing antigen at different concentrations with serum from a patient with high levels of antibody. Complexes were detectable over a 5 log range of antigen concentration. The lower limit of sensitivity of the assay was estimated to be 5.8 ng of complexed IgE per milliliter of patient serum. To determine if the assay could detect natural in vivo-formed complexes, sera from patients with positive RAST scores to egg white or milk were selected for a preliminary study. Three of the nine egg white-positive sera and two of the nine milk-positive sera were found to contain significant levels of IgE immune complexes. The versatility and sensitivity of the assay should now make it feasible to design experiments to elucidate the role, if any, of circulating immune complexes in the etiology of food allergy.


Surgical anatomy of the medial antebrachial cutaneous nerve.
In standard anatomy textbooks the course and distribution of the medial antebrachial cutaneous nerve and its branches are glossed over in a vague fashion as if they are of little importance. There are, however, clinical circumstances in which a knowledge of the anatomy of this nerve is invaluable. Sacrifice of the posterior branch in a medial approach to the elbow or cubital tunnel surgery can lead to annoying numbness over the olecranon and a symptomatic neuroma. The medial antebrachial cutaneous nerve is also frequently used in nerve grafting, especially in brachial plexus reconstruction, in which it is beneficial to know the available length and size of donor nerve. Fifty fresh cadaveric arms were dissected to define the course, distribution, size, and branches of the nerve.


Cultured Bloom's syndrome substrains: a relationship between growth in low serum and the expression of double minute chromosomes.
Four Bloom's syndrome fibroblast strains were examined for chromosome changes, immortalization, tumorigenicity, anchorage independent growth, and the ability to grow in low serum. Only one strain (GM 1492) exhibited some of these characteristics, which are generally associated with neoplastic cells. Strain GM 1492 also exhibited a low frequency of cells that contained double minute chromosomes. Substrains from GM 1492 were cloned, and showed that some of the above characteristics were expressed as a heritable trait. Karyotype instability (heteroploidy) was seen in all clones studied, with modal chromosome numbers ranging from near-diploid to near-tetraploid, as confirmed by DNA content distributions. Although no clones studied were tumorigenic in nude mice or immortalized, some showed an increased cellular proliferative capacity. Several clones were isolated after growth in 1% fetal bovine serum, and these all showed an increased expression of double minute chromosomes. Two of these 1% serum-isolated clones, (1492/clone 3-1% and clone 8-1%) were further studied by G-banding analysis, and found to show specific chromosomal anomalies. Clone 3 showed monosomy for chromosomes #4, #13, and #19, along with the absence of both copies of chromosome #7 but with both i(7p) and i(7q) chromosomes consistently seen. Clone 8 showed monosomy for chromosome #13. Preliminary experiments using DNA slot blots indicated that erb a and b, v-fes, v-mos, c-myc, n-myc, v-src, and v-sis showed no sequence amplification in GM 1492 cells or subclones.


[Early transjugular intrahepatic portosystemic shunt: When, how and in whom?].
Early TIPS is basically a new application of an old concept. This intervention used to be a useful rescue therapy when other interventions failed but has now become a primary intervention in patients with variceal bleeding and risk factors for poor prognosis. This technique has also been proven to control bleeding and has a definite survival advantage at 6 weeks and 1 year over standard therapy with vasoactive drugs and endoscopy, without increasing the rate of adverse events. In well-trained hands and with appropriate candidate selection, early TIPS is a safe, life-saving and evidenced-based procedure.


Trypsin-catalyzed synthesis of dipeptide containing alpha-aminoisobutyric acid using p- and m-(amidinomethyl)phenyl esters as acyl donor.
Two series of inverse substrates, p- and m-(amidinomethyl)phenyl esters derived from N-(tert-butyloxycarbonyl)amino acid, were prepared as acyl donor components for enzymatic peptide synthesis. They were found to be readily coupled with an acyl acceptor such as L-alanine p-nitroanilide to produce dipeptide. An alpha-aminoisobutyric acid containing dipeptide was especially obtained in satisfactory yield. Streptomyces griseus trypsin was a more efficient catalyst than the bovine trypsin. The optimum condition for the coupling reaction was studied by changing the organic solvent, pH, and acyl acceptor concentration. It was found that the enzymatic hydrolysis of the resulting product was negligible.


Effect of mirex, dechlorinated mirex derivatives and chlordecone on microsomal mixed-function oxidase activity and other hepatic parameters.
The effects of mirex, two monohydrogen and two dihydrogen mirex derivatives, and chlordecone on several hepatic parameters were studied 2 days following a single oral dose of 100 mg/kg in female rats. All compounds increased microsomal cytochrome P-450 content, NADPH-cytochrome c reductase activity, and hepatic ascorbic acid concentration. Microsomal protein concentration was generally increased. All compounds except chlordecone increased relative liver weight and the activities of aminopyrine N-demethylase and p-nitroanisole O-demethylase. Hepatic concentration of protein and glutathione were unaltered. The dechlorinated mirex derivatives caused effects of a magnitude similar to that of mirex, whereas chlordecone was considerably less potent.


Episodic-like and procedural memory impairments in histamine H1 Receptor knockout mice coincide with changes in acetylcholine esterase activity in the hippocampus and dopamine turnover in the cerebellum.
We investigated episodic-like (ELM) and procedural memory (PM) in histamine H1 receptor knockout (H1R-KO) mice. In order to relate possible behavioral deficits to neurobiological changes, we examined H1R-KO and wild-type (WT) mice in terms of acetylcholine esterase (AChE) activity in subregions of the hippocampus and AChE and tyrosine hydroxylase (TH) expression in the striatum. Furthermore, we analyzed acetylcholine (ACh), 5-HT and dopamine (DA) levels, including metabolites, in the cerebellum of H1R-KO and WT mice. The homozygous H1R-KO mice showed impaired ELM as compared with the heterozygous H1R-KO and WT mice. The performance of homozygous H1R-KO mice in the ELM task was primarily driven by familiarity-based memory processes. While the homozygous H1R-KO mice performed similar to the heterozygous H1R-KO and WT mice during the acquisition of a PM, as measured with an accelerating rotarod, after a retention interval of 7 days their performance was impaired relative to the heterozygous H1R-KO and WT mice. These findings suggest that, both, ELM and long-term PM are impaired in the homozygous H1R-KO mice. Neurochemical assays revealed that the H1R-KO mice had significantly lower levels of AChE activity in the dentate gyrus (DG) and CA1 subregions of the hippocampus as compared with the WT mice. The homozygous H1R-KO mice also displayed significantly reduced dihydroxyphenylacetic acid (DOPAC) levels and a reduced DOPAC/DA ratio in the cerebellum, suggesting that the DA turnover in the cerebellum is decelerated in homozygous H1R-KO mice. In conclusion, homozygous H1R-KO mice display severe long-term memory deficits in, both, ELM and PM, which coincide with changes in AChE activity in the hippocampus as well as DA turnover in the cerebellum. The importance of these findings for Alzheimer's (AD) and Parkinson's disease (PD) is discussed.


Histology obtained by needle biopsy gives additional information on the prognosis of hepatocellular carcinoma.
Hepatocellular carcinomas (HCC) have a strong biological heterogeneity. Current prognostic scores do not include histology. Information on the behavior of HCC based on histology has been characterized on retrospective data and large tissue specimens. We aimed to assess the additional value of needle biopsy and keratin 19 (K19) assessment in a prospective manner. Between 2003 and 2008, all patients with a confirmed diagnosis of HCC by a percutaneous or laparoscopic needle biopsy at the time of diagnosis, and of Barcelona Clinic Liver Cancer (BCLC) stage A, B or C, were included. The exclusion criterion was a palliative setting. Biopsies were scored for microvascular invasion, differentiation, K19, epithelial cell adhesion molecule and α-fetoprotein staining. Clinical and radiological features were registered at time of biopsy. The added value of K19 was assessed using Cox proportional hazards regression. Of 74 patients screened, we included 58 patients. Based on the BCLC, 41% presented with early disease (BCLC A), 16% with intermediate disease (BCLC B) and 43% with advanced disease (BCLC C). In nine patients (16%), K19 staining was positive. Median follow up was 54 months (range 1-74) and 43 patients (72%) died. BCLC classification predicted the prognosis accurately, but histology offered additional prognostic information. In multivariate analysis, K19 was a strong predictor of overall survival (hazard ratio 4.57, 95% confidence interval 1.86-10.6), which improved predictive performance. No needle tract dissemination was observed. Despite the possible problem of sampling error, needle biopsy offered additional prognostic information. This is especially the case for K19 staining.


A possible role of proopiomelanocortin peptides in self-injurious behavior.
1. The hypothesis that opioids may be involved in self-injurious behavior is supported by fifteen years of basic animal research suggesting that opioid peptides of the brain and spinal cord participate in the modulation of antinociception in animals, and research of animal models for self-injurious behavior utilizing exogenously administered opiate agonists. 2. Clinical biochemical and pharmacological research conducted over the past five years has also suggested the possibility that opioid peptides may play an important etiological role in the elaboration of self-injurious behavior in some individuals. 3. An opioid overactivity self-injurious hypothesis is supported by results of one study indicating elevated Fraction II opioids (enkephalins) in the lumbar-thecal cerebrospinal fluid of self-injurious children compared to controls, and by the five out of six published studies demonstrating statistically significant decreases in the frequency of self-injurious behavior with the opiate antagonist, naltrexone. 4. A very recent investigation has suggested that some self-injurious individuals show abnormalities in their plasma proopiomelanocortin peptide response to naltrexone, thereby indicating a possible dysfunction in the responsitivity of the proopiomelanocortin system of the hypothalamic-pituitary-adrenal axis of these individuals. 5. These data and results of other investigations have resulted in the elaboration of the original opioid hypothesis to a more comprehensive biochemical model that focuses on this proopiomelanocortin dysregulation. 6. Because of biochemical and functional interrelationships between proopiomelanocortin peptides and certain other neurochemical systems in the brain, it is proposed that pineal melatonin and serotonin may also be involved in this dysregulation. Further basic and clinical research will be needed to test the proposed biochemical model for self-injurious behavior.


Polyelectrolyte-modified short microchannel for cation separation.
Three alkali cations, potassium, sodium, and lithium, have been separated within 15 s in a 1 cm long polymer microchip. The separation microchannel is modified by a polycation, poly(allylammonium chloride), which makes the channel surfaces positively charged leading to a reversed electroosmotic flow (EOF) when compared to bare channels. Due to the decreased apparent mobility of the cations, the separation resolution is improved allowing the use of shorter channels.


On the possibility that opiate and ethanol actions are mediated by similar mechanisms.
Opiates and alcohol modify neuronal electrical activity in many sites of the nervous system. Both act as depressants or stimulants of cell firing depending upon the type of cell studied. Alcohol seems to act upon some nerves by changing their membrane ionic conductance, while opiates seem to affect synaptic events. However, all of the known neural actions of these substances involve calcium-dependent mechanisms. A hypothesis is proposed to account for these facts.


The high performance liquid chromatography electrospray ionization mass spectrometry analysis of diverse basic pharmaceuticals on cyanopropyl and pentafluorophenylpropyl stationary phases.
Cyanopropyl (CN) and pentafluorophenylpropyl (PFPP) modified silica columns give good retention and good peak shape for the high performance liquid chromatography/electrospray ionization/mass spectrometry (HPLC/ESI/MS) analysis of several classes of basic drugs. These phases enhance the ESI-MS signal by providing good retention of basic drugs with a mobile phase containing 90% acetonitrile. With C18 columns, in order to achieve good retention of basic drugs, only a mobile phase containing less than 40% acetonitrile can be used. Higher concentrations of acetonitrile produce a larger MS signal in the ESI process; the MS signal was a factor of 9 and 12 times greater on the CN and PFPP phases when compared with the C18 phase for the analysis of codeine. The C18 phase required only 4.0-6.0% acetonitrile to obtain the same retention time for codeine. The CN and PFPP stationary phases can be used for the analysis of a range of basic drugs, including many compounds which are poorly retained on the popular C18 and C8 stationary phases. Applications of CN and PFPP columns in the HPLC/ESI/MS of basic drugs include the analysis of antimalarials, such as quinine, bronchodilators, such as salbutamol and tulobuterol, cardioactive drugs, such as procainamide and beta-blockers, tricyclic antidepressants (TCAs), such as protriptyline and trimipramine and alkaloids, such as morphine and codeine. The CN and PFPP phases are also useful for the analysis of bufuralol and its metabolite, hydroxy-bufuralol. All the above analyses were performed using the same mobile phase, 90% acetonitrile; thus the HPLC method development process was expedited. The CN and PFPP phases also gave reproducible retention times and peak shape after more than 8 h of analyses.


Innovations in proteomic profiling of cancers: alternative splice variants as a new class of cancer biomarker candidates and bridging of proteomics with structural biology.
Alternative splicing allows a single gene to generate multiple RNA transcripts which can be translated into functionally diverse protein isoforms. Current knowledge of splicing is derived mainly from RNA transcripts, with very little known about the expression level, 3D structures, and functional differences of the proteins. Splicing is a remarkable phenomenon of molecular and biological evolution. Studies which simply report up-regulation or down-regulation of protein or mRNA expression are confounded by the effects of mixtures of these isoforms. Besides understanding the net biological effects of the mixtures, we may be able to develop biomarker tests based on the observable differential expression of particular splice variants or combinations of splice variants in specific disease states. Here we review our work on differential expression of splice variant proteins in cancers and the feasibility of integrating proteomic analysis with structure-based conformational predictions of the differences between such isoforms.


Diagnosis and relining techniques for delayed type IIIB endoleaks with the second-generation AFX endograft.
Type IIIB endoleaks resulting from endograft fabric tear are an uncommon but serious late complication of endovascular aortic aneurysm repair. The Strata fabric used in the earlier generation AFX endograft (updated to Duraply in October 2014) has been associated with an increased frequency of these events. Herein we report on two patients exhibiting delayed type IIIB endoleaks after AFX device insertion to treat an abdominal aortic aneurysm and discuss optimal relining techniques.


Coverage of community case management for malaria through CHWs: a quantitative assessment using primary household surveys of high-burden areas in Chhattisgarh state of India.
Community Case Management of Malaria (CCMM) has been implemented through community health workers (CHWs) in many countries. Existing studies have shown that CHWs can be viable means of implementing CCMM. However, not many studies have examined the coverage under large-scale CCMM programmes. India is a big contributor to global malaria burden. Chhattisgarh is a leading state in India in terms of malaria incidence and mortality. CCMM was implemented on a large scale through the 'mitanin' CHWs in rural Chhattisgarh from 2015. Under CCMM, 37,696 CHWs in 84 high-burden administrative blocks of the state were trained and equipped with rapid diagnostic tests (RDT), artemisinin-based combination therapy (ACT) and chloroquine. This descriptive quantitative study assesses coverage of CCMM in detection and treatment of Malaria over three rounds of household surveys-2015, 2016 and 2018. Household-interviews covered more than 15,000 individuals in each round, using multi-stage random sampling across the 84 blocks. The main objectives were to find out the coverage in identification and treatment of malaria and the share of CHWs in them. A 15-days recall was used to find out cases of fever and healthcare sought by them. In 2018, 62% of febrile cases in rural population contacted CHWs. RDT, ACT and chloroquine were available with 96%, 80% and 95% of CHWs, respectively. From 2015 to 2018, the share of CHWs in testing of febrile cases increased from 34 to 70%, while it increased from 28 to 69% in treatment of malaria cases. CHWs performed better than other providers in treatment-completion and administered medication under direct observation to 72% of cases they treated. This study adds to one of the most crucial but relatively less reported area of CCMM programmes, i.e. the extent of coverage of the total febrile population by CHWs, which subsequently determines the actual coverage of case-management in malaria. Mitanin-CHWs achieved high coverage and treatment-completion rates that were rarely reported in context of large-scale CCMM elsewhere. Close to community, well-trained CHWs with sufficient supplies of rapid tests and anti-malarial drugs can play a key role in achieving the desired coverage in malaria-management.


Early findings and strategies for successful implementation of SIMPL workplace-based assessments within vascular surgery residency and fellowship programs.
As medical education systems increasingly move toward competency-based training, it is important to understand the tools available to assess competency and how these tools are utilized. The Society for Improving Medical Professional Learning (SIMPL) offers a smart phone-based assessment system that supports workplace-based assessment of residents' and fellows' operative autonomy, performance, and case complexity. The purpose of this study was to characterize implementation of the SIMPL app within vascular surgery integrated residency (0+5) and fellowship (5+2) training programs. SIMPL operative ratings recorded between 2018 and 2022 were collected from all participating vascular surgery training institutions (n = 9 institutions with 5+2 and 0+5 programs; n = 4 institutions with 5+2 program only). The characteristics of programs, trainees, faculty, and SIMPL operative assessments were evaluated using descriptive statistics. Operative assessments were completed for 2457 cases by 85 attendings and 86 trainees, totaling 4615 unique operative assessment ratings. Attendings included dictated feedback in 52% of assessments. Senior-level residents received more assessments than junior-level residents (postgraduate year [PGY]1-3, n = 439; PGY4-5, n = 551). Performance ratings demonstrated increases from junior to senior trainees for both resident and fellow cohorts with "performance-ready" or "exceptional performance" ratings increasing by nearly two-fold for PGY1 to PGY5 residents (28.1% vs 40.6%), and from first- to second-year fellows (PGY6, 46.7%; PGY7, 60.3%). Similar gains in autonomy were demonstrated as trainees progressed through training. Senior residents were more frequently granted autonomy with "supervision only" than junior residents (PGY1, 8.7%; PGY5, 21.6%). "Supervision only" autonomy ratings were granted to 21.8% of graduating fellows. Assessment data included a greater proportion of complex cases for senior compared with junior fellows (PGY6, 20.9% vs PGY7, 26.5%). Program Directors felt that faculty and trainee buy-in were the main barriers to implementation of the SIMPL assessment app. This is the first description of the SIMPL app as an operative assessment tool within vascular surgery that has been successfully implemented in both residency and fellowship programs. The assessment data demonstrates expected progressive gains in trainees' autonomy and performance, as well as increasing case complexity, across PGY years. Given the selection of SIMPL as the assessment platform for required American Board of Surgery and Vascular Surgery Board Entrustable Professional Activities assessments, understanding facilitators and barriers to implementation of workplace-based assessments using this app is imperative, particularly as we move toward competency-based medical education.


Optimization of carboxymethylcellulase production from Bacillus amyloliquefaciens SS35.
In this paper, we have attempted optimization of production of enzyme carboxymethylcellulase or endoglucanase from the bacterium Bacillus amyloliquefaciens SS35. Optimization has been carried out in two stages using statistical experimental design, viz. medium optimization and optimization of fermentation parameters. For medium optimization, Plackett-Burman design followed by central composite design (CCD) was used, while for optimization of fermentation parameters one-variable-at-a-time method followed by CCD was used. Carbon and nitrogen sources in the medium have been revealed to be the significant factors for enzyme production (carboxymethylcellulose 18.05 g/L; yeast extract 8 g/L and peptone 2 g/L). The inorganic salts have been found to be insignificant components of medium. Optimum fermentation parameters for optimized medium were: initial medium pH 5.65, incubation temperature = 40 °C, shaking speed = 120 rpm, and inoculum size = 6.96 %, v/v. Interestingly, the influence of all four parameters was almost independent with no interlinks. Secondly, the overall effect of all parameters was also low, as indicated by linear, square and interaction regression coefficients that were at least one order of magnitude lower than the intercept in the model equation. These results essentially meant that medium components dominate overall enzyme production process in comparison to fermentation parameters.


TGFβ3 (TGFB3) polymorphism is associated with male infertility.
Factors affecting the blood-testis barrier function may be involved in testicular damage and male infertility. Two cytokines play an important role in the barrier regulation, namely transforming growth factor beta 3 (TGF-β3) and tumor necrosis factor (TNF-α). The aim of this study was to investigate the potential association between TGF-β3 (TGFB3) and TNF-α (TNF) gene polymorphisms and male infertility. A total of 846 subjects, 423 diagnosed with male infertility and 423 fertile men were enrolled. TGFB3 (rs2268626:T > C, rs3917158:C > T, rs2284792:A > G, rs2268625:T > C, rs3917187:C > T) and TNF (rs1800629:-308G > A) gene polymorphisms were genotyped. No association between TNF genotype and infertility was observed. As for TGFB3, the genotypes distribution was similar in infertile and fertile men. However, rs2284792 minor allele frequency was significantly higher among infertile subjects. Heterozygous rs2284792 AG genotype was associated with increased odds for infertility [OR = 1.40 (95% CI 1.05-1.86), p = 0.021] and similar results were observed for G allele carrier status [OR = 1.40 (95% CI 1.06-1.84), p = 0.017]. Heterozygosity in TGFB3 rs3917158 was also associated with the infertility [OR = 1.37 (95% CI 1.01-1.87), p = 0.041]. The TGFB3 variant genotypes were associated with lower spermatozoa motility parameters in fertile men. The results indicate that variants in TGFB3 gene may be associated with male infertility. However, the findings require further replication and validation.


The role of neutrophil elastase in aortic valve calcification.
Calcific aortic valve disease (CAVD) is the most commonly valvular disease in the western countries initiated by inflammation and abnormal calcium deposition. Currently, there is no clinical drug for CAVD. Neutrophil elastase (NE) plays a causal role in inflammation and participates actively in cardiovascular diseases. However, the effect of NE on valve calcification remains unclear. So we next explore whether it is involved in valve calcification and the molecular mechanisms involved. NE expression and activity in calcific aortic valve stenosis (CAVD) patients (n = 58) and healthy patients (n = 30) were measured by enzyme-linked immunosorbent assay (ELISA), western blot and immunohistochemistry (IHC). Porcine aortic valve interstitial cells (pVICs) were isolated and used in vitro expriments. The effects of NE on pVICs inflammation, apoptosis and calcification were detected by TUNEL assay, MTT assay, reverse transcription polymerase chain reaction (RT-PCR) and western blot. The effects of NE knockdown and NE activity inhibitor Alvelestat on pVICs inflammation, apoptosis and calcification under osteogenic medium induction were also detected by RT-PCR, western blot, alkaline phosphatase staining and alizarin red staining. Changes of Intracellular signaling pathways after NE treatment were measured by western blot. Apolipoprotein E-/- (APOE-/-) mice were employed in this study to establish the important role of Alvelestat in valve calcification. HE was used to detected the thickness of valve. IHC was used to detected the NE and α-SMA expression in APOE-/- mice. Echocardiography was employed to assess the heat function of APOE-/- mice. The level and activity of NE were evaluated in patients with CAVD and calcified valve tissues. NE promoted inflammation, apoptosis and phenotype transition in pVICs in the presence or absence of osteogenic medium. Under osteogenic medium induction, NE silencing or NE inhibitor Alvelestat both suppressed the osteogenic differentiation of pVICs. Mechanically, NE played its role in promoting osteogenic differentiation of pVICs by activating the NF-κB and AKT signaling pathway. Alvelestat alleviated valve thickening and decreased the expression of NE and α-SMA in western diet-induced APOE-/- mice. Alvelestat also reduced NE activity and partially improved the heart function of APOE-/-mice. Collectively, NE is highly involved in the pathogenesis of valve calcification. Targeting NE such as Alvelestat may be a potential treatment for CAVD.


Targeting megalin to enhance delivery of anti-clusterin small-interfering RNA nanomedicine to chemo-treated breast cancer.
The goal of this study is to evaluate a new targeting strategy to improve nanomedicine delivery to breast cancer cells that survive prior exposure to chemotherapy. These cells are particularly difficult to treat because they often develop drug resistance by upregulation of chemoresistant factors such as clusterin and should be preferably eradicated before they further spread out. In this study, the surface endocytotic receptor megalin was studied for the first time for targeted delivery of anti-clusterin small-interfering RNAs (siRNAs) to these chemo-treated cells. Lipid-polyethylenimine hybrid nanocarriers decorated with apolipoprotein E (Ap-LPNs) were developed for this purpose. Using immunoblotting, we demonstrated induction of both megalin and clusterin in MCF-7 cells by previous paclitaxel treatment. The siRNA transfection of these megalin-rich chemo-treated cancer cells was improved by three-fold when the siRNAs were delivered by Ap-LPNs. This trend was translatable into enhanced clusterin knockdown and improved chemosensitization to subsequent paclitaxel treatment (both p<0.05 versus uncoated LPNs). This proof-of-principle study has validated a novel "chemoresistance-targeting" strategy for siRNA delivery to the cancer cell subpopulation that begins to acquire chemoresistance and is in strong need for chemosensitization.


Mercury toxicology as assessed through fish scales.
Increasing pollution in water bodies is directly or indirectly related to increasing urbanization and indiscriminate disposal of agrochemical & industrial effluents. Heavy metals are one of the important categories of such pollutants and are causing serious hazards to non target species (Mwachiro and Druve1997). Their salts constitute a serious type of pollution in fresh water and being stable compounds; they are not readily removed by oxidation, precipitation or other processes and affect the activity in recipient animal (Jagadeesan and Vijayalakshmi 1998). Increase in the concentration of heavy metals has been reported in water of Vasai Creek, Maharastra and surface as well as groundwater of Delhi (Lokhande and Kelker 1999; Dixit et al. 2003). Recently, Food and Drug Adminstration (USFDA) and Environmental Protection Agency (USEPA) (2004) in their combined report recommended that pregnant women and lactating mothers should not eat shark, sword fish and king Mackerel as these fishes contain high levels of mercury.


The medical and dental attendance pattern of patients attending general dental practices in Warwickshire and their general health risk assessment.
The dental team could have an important role to play in general health risk assessment within primary and community healthcare.Aims To describe medical and dental attendance patterns, demographics and health profiles of patients routinely attending general dental practices in Warwickshire. To identify whether a subgroup attend dental practices regularly but attend medical practices infrequently and discuss whether preventive healthcare interventions delivered in general dental practice would be appropriate. A self-completion questionnaire was administered to patients attending 16 dental practices in Warwickshire. Eight hundred and eleven completed questionnaires were returned (74% response). Seven hundred and eighty-nine (98%) respondents visited their dentist every one to two years or more frequently and of these a subgroup of 121 (15.3%) visited their general medical practice surgery or health centre less often than every two years. In the subgroup 9.5% reported high blood pressure, 17.6% currently smoked, 22% drank above recommended guidelines, 32.1% were overweight and 7.3% obese. The data suggest there may be a role for dental practitioners in identifying patients at risk of having undiagnosed or future general health problems and providing appropriate general health advice, screening or signposting the patient to relevant general healthcare facilities either within or external to the dental practice.


Absolute temperature, temperature changes and stroke risk: a case-crossover study.
Studies suggest that there is an association between weather patterns and ischemic stroke risk. Exposure to a sudden decrease in temperature may increase stroke risk through altering blood viscosity and/or by triggering infections. We investigated the association between ischemic stroke risk and change in temperature. We used a case-crossover study design with 303 consecutive patients admitted to Heidelberg University, Department of Neurology over a one and a half year period (Aug 1998-Jan 2000). We used one day before stroke as the hazard (case) period matched to two control periods 2-7 days before and after stroke onset and took both ambient maximum temperature and the 24-hour difference in maximum temperature as exposure. There was no risk associated with ambient maximum temperature at all lag times and in all subgroup analyses. For the 24-hour difference, large changes in temperature (>5 degrees C) were associated with an increased risk of acute ischemic stroke regardless of whether the change was negative or positive. The odds ratio for temperature increases >5 degrees C compared to no change in temperature was 2.0 (95% confidence interval (CI): 0.7-5.9) at a lag time of 3 days. We found no relevant relation between temperature and stroke risk. The results suggest that the risk of ischemic stroke may increase with large day-to-day variations upwards or downwards in temperature.


[What is confirmed in the treatment of chronic inflammatory bowel diseases].
The prevalence of the chronic inflammatory bowel diseases (CIBD) Crohn's disease (CD) and ulcerative colitis (UC) is on the rise worldwide. In Germany CIBDs are also a significant healthcare problem. The pathogenesis is complex and involves genetic factors, environmental aspects and changes in the immunological constitution. Furthermore, the gut microbiota plays a role in the maintenance of intestinal inflammation. Fortunately, several new drugs, in particular biologicals, have been approved for the treatment of CIBDs. The treatment of UC is mainly based on 5‑aminosalicylic acid formulations, preferably as a topical form for distal colitis and proctitis as well as local budesonide formulations. In the case of extensive spread, high disease activity and refractory disease antibodies (biologicals) are successfully used, similar to CD. In addition to anti-tumor necrosis factor antibodies (infliximab, adalimumab, golimumab), vedolizumab, an anti-integrin antibody and the interleukin 12/23 antibody ustekinumab can be successfully used. The intravenous and also subcutaneous administration of antibodies are increasing in importance and are now available for all forms. Furthermore, the Janus kinase inhibitor tofacitinib is an orally administered option for UC. Clinical scores, endoscopy, ultrasound, laboratory parameters and calprotectin determination in stool are employed to evaluate treatment response (treat to target approach). Ultimately, the long-term goal is mucosal healing. Despite advances in the pharmaceutical treatment, a significant number of patients with CIBD still suffer from treatment refractory courses and need surgery at some time during the disease.


Extreme Vocal Effects Distortion, Growl, Grunt, Rattle, and Creaking as Measured by Electroglottography and Acoustics in 32 Healthy Professional Singers.
Vocal effects - also called extreme or extended vocal techniques - with the intention to sound hoarse or rough are widely used as part of many genres and styles of singing, yet scarcely documented in research. Physiological studies detail the involvement of supraglottic structures for the production of vocal effects, yet the acoustic impact of such involvement has not been documented systematically across phonation types. To report acoustic measurements and electroglottography-specific measurements for the five rough-sounding vocal effects Distortion, Growl, Grunt, Rattle, and Creaking across phonation types to demonstrate differences between notes with and without vocal effects added. Thirty-two professional singers and singing teachers produced sustained vowels in each of the four vocal modes with alternations of adding and removing the vocal effects. The singers were recorded with a microphone at a constant distance as well as with EGG. The vocal effects Distortion, Growl, Grunt, Rattle, and Creaking impact the acoustic spectra in separate and systematic ways across genders and phonation types. Each vocal effect impacted the spectrum in specific and particular frequency regions between 0 and 3.5 KHz as well as in higher partials after 12 kHz with statistical significance. EGG-waveforms were un-impacted by most of the vocal effects produced using supraglottic sound sources, whereas Grunt and Creaking conditions did impact EGG-waveform signals, though not consistently between participants. EGG measures confirmed sustained and unchanged Qx and Fx for most conditions, with statically significant changes in noise measurements Harmonic-to-Noise Ratio, Normalised Noise Energy, Relative Average Perturbation, and Cepstral Peak Prominence, despite Sound Pressure Level differing significantly only for a few specific conditions. Singers scored an average of 5,95 on Voice Handicap Index questionnaires and were all reportedly healthy. Vocal effects added to phonation produce specific increases and specific decreases in particular frequency regions in a systematic way and can be produced in a healthy and sustainable manner, as measured by Voice Handicap Index. Vocal effects can be added to different phonation types with differing acoustic output and singers were able to sustain and control involvement of the supraglottic sound source(s) independently of phonation type.


First trimester screening for Down syndrome in rhesus negative women.
To explore the effect of maternal rhesus status on first-trimester screening markers for Down syndrome. We accessed a database of singleton pregnancies undergoing first-trimester genetic screen with maternal Rh status documented and pregnancy outcome information available. Excluded were cases of fetal chromosomal or structural abnormalities, or maternal systemic disease. Results of maternal serum pregnancy-associated plasma protein A (PAPP-A) and beta-human chorionic gonadotrophin (beta-hCG) adjusted for gestational age were compared between Rh-negative and Rh-positive women with p < 0.05 considered significant. Two thousand two hundred and two pregnancies fulfilled the study criteria, and 160 of them (7%) were Rh negative. Only free beta-hCG corrected multiples of the median (MoM) values were statistically increased in Rh-negative women (p < 0.009). Using a cut-off of 1:300, screen-positive rates of maternal serum biochemistry were not significantly different between Rh-negative and Rh-positive women (12.5 vs 10.4%, p = 0.41). The present study focused on measurements of beta-hCG and PAPP-A in the sera of women with Rh-negative blood group. Women with Rh-negative blood type have similar first-trimester serum PAPP-A MoM values as Rh-positive women, but significantly higher beta-hCG MoM values. However, there was no significant difference in the screen-positive rate for Down syndrome between the two groups.


[Chemokine receptor CXCR4 gene silencing with shRNA inhibits breast cancer metastasis to the lung in nude mice].
To construct a CXCR4 specific recombinant plasmid vector and study its inhibiting effect on invasion capacity in vitro of human breast cancer MDA-MB-231 cell line and its metastatic potential to the lung in nude mice. A CXCR4 specific recombinant plasmid vector was constructed and transfected into the cultured MDA-MB-231 cell line with lipofectamine 2000. RT-PCR and Western blot were used to detect the mRNA and protein expression of CXCR4, respectively. Invasion capability in vitro of the cells was evaluated by Boyden chamber. The cell proliferation capacity was detected by MTT method. The nude mouse model of lung metastasis was established by injection of MDA-MB-231 cells into the tail vein. The animals were sacrificed at 6 weeks after the tumor cells injection. Whole lung tissues were harvested, embedded in paraffin, sectioned serially, and the HE-stained paraffin sections were examined pathologically to evaluate the presence and number of metastatic tumors. The CXCR4 mRNA expression rate was 29.5% +/- 3.8% in the CXCR4-shRNA group, significantly lower than that of the control group (69.7% +/- 2.6%, P < 0.01) and mock-control group (67.8% +/- 3.5%, P < 0.01). The CXCR4 protein expression rate was 15.4% +/- 1.1% in the CXCR4-shRNA group, significantly lower than that of the control group (39.0% +/- 2.4%, P < 0.01) and mock-control group (35.9% +/- 3.9%, P < 0.01). Silencing of CXCR4 by shRNA lead to a significant decrease in breast cancer cell invasion and proliferation capacity in vitro. Furthermore, tumor cells with CXCR4 shRNA permanent transfcetion had a much lower lung metastatic potential in nude mice than control cells and mock control cells in vivo. CXCR4 shRNA can inhibit the expression of CXCR4 and decrease the invasion and lung metastatic potential of human breast cancer cells.


Pre-operative visits by ITU nurses: recommendations for practice.
Pre-operative visiting by theatre nurses is now policy in many hospitals, following an explosion of research studies outlining the benefits to the patient. However, the author could find very little available literature on pre-operative visiting by intensive therapy unit (ITU) nurses, to patients electively admitted to ITU following surgery. The purpose of this project is to explore the need for patient information, outline the aims of an ITU nurse's pre-operative visit, and discuss the timing of the proposed visit, the information to give the patient, the inclusion of the patient's family and the provision of printed literature. Finally, recommendations are made for implementing a pre-operative visiting service by ITU nurses.


Orthostatic hypotension from sympathetic denervation in Parkinson's disease.
Patients with PD often have signs or symptoms of autonomic failure, including orthostatic hypotension. Cardiac sympathetic denervation occurs frequently in PD, but this has been thought to occur independently of autonomic failure. Forty-one patients with PD (18 with and 23 without orthostatic hypotension) and 16 age-matched healthy volunteers underwent PET scanning to visualize sympathetic innervation after injection of 6-[(18)F]fluorodopamine. Beat-to-beat blood pressure responses to the Valsalva maneuver were used to identify sympathetic neurocirculatory failure and plasma norepinephrine to indicate overall sympathetic innervation. All patients with PD and orthostatic hypotension had abnormal blood pressure responses to the Valsalva maneuver and septal and lateral ventricular myocardial concentrations of 6-[(18)F]fluorodopamine-derived radioactivity >2 SD below the normal mean. In contrast, only 6 of the 23 patients without orthostatic hypotension had abnormal Valsalva responses (p < 0.0001 compared with patients with orthostatic hypotension), and only 11 had diffusely decreased 6-[(18)F]fluorodopamine-derived radioactivity in the left ventricular myocardium (p = 0.0004). Of the 12 remaining patients without orthostatic hypotension, 7 had locally decreased myocardial radioactivity. Supine plasma norepinephrine was lower in patients with than in those without orthostatic hypotension (1.40 +/- 0.15 vs 2.32 +/- 0.26 nmol/L, p = 0.005). 6-[(18)F]fluorodopamine-derived radioactivity was less not only in the myocardium but also in the thyroid and renal cortex of patients with PD than in healthy control subjects. In PD, orthostatic hypotension reflects sympathetic neurocirculatory failure from generalized sympathetic denervation.


Pulmonary complications of the acquired immunodeficiency syndrome: a clinicopathologic study of 70 cases.
The pulmonary complications of 70 patients with the acquired immunodeficiency syndrome (AIDS) are reviewed. Pneumocystis carinii pneumonia (PCP), present in 67 per cent of the patients, was diagnosed by fiberoptic bronchoscopy with transbronchial biopsies in all of the patients except two adults, who required open lung biopsy, and two children, in whom the infection was detected only at autopsy. Other opportunistic infections, such as cytomegalovirus pneumonitis, mycobacterial infections, invasive candidiasis, toxoplasmosis, cryptococcosis, and histoplasmosis, were more difficult to diagnose by fiberoptic bronchoscopy. In only four cases were these conditions detected during life. Neoplasms and lymphoproliferative processes also presented diagnostic problems, and only one case each of Kaposi's sarcoma and lymphoid interstitial pneumonitis were detected by fiberoptic bronchoscopy. In four other cases these conditions, as well as two pulmonary lymphomas, diffuse large cell immunoblastic type, were detected only at autopsy. Sixty-eight per cent of the patients in this study died, usually with progressive intractable respiratory failure and pulmonary complications that had not been diagnosed during life, including potentially treatable diseases, such as bacterial pneumonias, PCP, nontuberculous mycobacteria, invasive candidiasis, toxoplasmosis, and invasive aspergillosis. The need for earlier detection of pulmonary complications in patients with AIDS is discussed.


[In vitro study of electrospun polycaprolactone/chitosan nano fiber membrane guided bone regeneration].
To explore the effect of electrospining chitosan/polycaprolactone nano fiber membrane on regeneration of bone defects in vitro. Chitosan/polycaprolactone nanofiber membrane was prepared by electrospinning technique. According to alkaline phosphatase activity and Western blot results, the ability of osteogenic differentiation of cells by fiber membrane as carrier was evaluated. The data was analyzed statistically with SPSS17.0 software package. The osteogenic differentiation capacity of electrospining chitosan/polycaprolactone nano fiber membrane was significantly higher than the control (without materials), and the difference was significant (P<0.05). Electrospinning chitosan/polycaprolactone nano fiber membrane can promote osteogenic differentiation of cells in vitro, which provides a scientific basis for clinical regeneration of bone defects.


Clawing back: broadening the notion of metal chelators in medicine.
The traditional notion of chelation therapy is the administration of a chemical agent to remove metals from the body. But formation of a metal-chelate can have biological ramifications that are much broader than metal elimination. Exploring these other possibilities could lead to pharmacological interventions that alter the concentration, distribution, or reactivity of metals in targeted ways for therapeutic benefit. This review highlights recent examples that showcase four general strategies of using principles of metal chelation in medicinal contexts beyond the traditional notion of chelation therapy. These strategies include altering metal biodistribution, inhibiting specific metalloenzymes associated with disease, enhancing the reactivity of a metal complex to promote cytotoxicity, and conversely, passivating the reactivity of metals by site-activated chelation to prevent cytotoxicity.


Sex-related difference in substance P level in rat anterior pituitary: a model of neonatal imprinting by testosterone.
There was a distinct sex-related difference in the anterior pituitary level of substance P-like immunoreactivity (SP-LI) in adult rats; the anterior pituitaries of male rats contained a higher concentration of SP-LI than those of females. Neonatal castration (day 1) resulted in a marked decrease of the SP-LI level in adult males, and this decrease in the SP-LI level was restored by neonatal testosterone replacement (days 2, 4, 6) but not by androgen replacement in adulthood. On the other hand, testosterone injection of neonatal females (days 2, 4, 6) caused a significant elevation of the SP-LI level after maturation. Ovariectomy of adult females failed to alter the SP-LI level. These results indicate that neonatal exposure to testosterone plays a critical role in the sexual differentiation of the substance P level in rat anterior pituitary.


Activation of ATM and phosphorylation of p53 by heat shock.
p53 protein is phosphorylated in response to various stresses. Here we examined phosphorylation of p53 protein in normal human diploid cells after heat shock at 43 degrees C for 2 h. We found that heat shock stimulates phosphorylation of p53 at Ser15 but not at Ser20, while X-irradiation at 4 Gy and 10 J/m(2) of UV induces phosphorylation of p53 at Ser15 and less significantly at Ser20. Increased phosphorylation of Ser15 was also observed in heat shocked GM638, the SV40-transformed human fibroblast cell line. Although X-ray irradiation induced phosphorylation of Ser6, 9, 20, and 37 in GM638 cells, heat shock did not affect the phosphorylation level of these serines. We observed little or no phosphorylation of p53 at Ser15 in two primary ataxia telangiectasia fibroblast cells, that are defective in ATM. Using an in vitro kinase assay, we confirmed that immunoprecipitated ATM from both heat-shocked and X-irradiated normal human diploid cells can phosphorylate p53 at Ser15 to a similar extent. These results indicate that heat shock induces phosphorylation of p53, especially at Ser15, and its phosphorylation is mediated by ATM kinase.


A novel method for salts removal from municipal solid waste incineration fly ash through the molten salt thermal treatment.
The disposal of the hazardous municipal solid waste (MSW) incineration fly ash is a critical environmental issue in China and the high contents of salts in the fly ash make the ash disposal extremely difficult. The present study proposes a novel method for the salts removal from MSW incineration fly ash using molten carbonates and chlorides at moderate temperatures from 773 K to 1073 K. The results showed that molten salts could effectively extract alkali and alkaline earth metals chlorides and sulfates from the fly ash. Other ash components, like Si/Al-compounds, were precipitated from the molten salts and concentrated in residues. By comparison, molten carbonates showed greater capability in the salts extraction while molten chlorides showed better selectivity in chlorides removal from MSW incineration fly ash. These findings suggest that the optimization of molten salts system could further prove the potential applicability of molten salts thermal treatment method for the salts removal from MSW incineration fly ash.


Complete assignments of 1H and 13C NMR spectral data for three new triterpenoid saponins from Ilex hainanensis Merr.
Three new oleanane-type triterpenoid saponins, ilexhainanoside C, D and E, all with 24, 28-dioic acid groups, were isolated from the leaves of Ilex hainanensis. They were 3beta-hydroxyolean-12-ene-24, 28-dioic acid-28-O-beta-D-glucopyranoside(1), 3beta, 19alpha-dihydroxyolean-12-ene-24, 28-dioic acid-28-O-beta-D-glucopyranoside(2) and 3beta, 29-dihydroxyolean-12-ene-24, 28-dioic acid-28-O-beta-D-glucopyranoside(3). The structures of these three new compounds were elucidated and complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments [heteronuclear single quantum coherence (HSQC), HMBC and rotational nuclear Overhauser effect spectroscopy (ROESY)].


Radiographic changes differ between two different short press-fit humeral stem designs in total shoulder arthroplasty.
The purpose of this study was to compare the radiographic changes of the humerus in the short term after total shoulder arthroplasty with two different short-stem humeral components. The hypothesis was that there would be no difference in radiographic changes or functional outcome based on component type. A retrospective review was conducted of primary total shoulder arthroplasties performed with a short press-fit humeral component. Group A included a collarless humeral stem with an oval geometry and curved stem (Ascend or Ascend Flex; Wright Medical, Memphis, TN, USA). Group B included a humeral stem with a metaphyseal collar, rectangular geometry, and straight stem (Apex; Arthrex, Inc., Naples, FL, USA). Radiographic changes and functional outcome were evaluated at a minimum of 2 years postoperatively. There were 42 patients in group A and 35 patients in group B available for analysis. There was no difference in functional outcome between the groups. In group A, the mean total radiographic change score of the humerus was 3.9, with changes classified as low in 38% and high in 62%. In group B, the mean total radiographic change score of the humerus was 2.5, with changes classified as low in 77% and high in 23% (P < .001). Medial calcar osteolysis was present in 71% of group A compared with 28.5% of group B (P < .001). At short-term follow-up, there is no difference in functional outcome or revision between 2 different humeral stem designs. However, bone adaptive changes and the rate of medial calcar osteolysis are significantly different.


NR1D1 Deletion Induces Rupture-Prone Vulnerable Plaques by Regulating Macrophage Pyroptosis via the NF-κB/NLRP3 Inflammasome Pathway.
Vulnerable plaque rupture is the main trigger of most acute cardiovascular events. But the underlying mechanisms responsible for the transition from stable to vulnerable plaque remain largely unknown. Nuclear receptor subfamily 1 group D member 1 (NR1D1), also known as REV-ERB α, is a nuclear receptor that has shown the protective role in cardiovascular system. However, the effect of NR1D1 on vulnerable plaque rupture and its underlying mechanisms are still unclear. By generating the rupture-prone vulnerable plaque model in hypercholesterolemic ApoE-/- mice and NR1D1-/-ApoE-/- mice, we demonstrated that NR1D1 deficiency significantly augmented plaque vulnerability/rupture, with higher incidence of intraplaque hemorrhage (78.26% vs. 47.82%, P = 0.0325) and spontaneous plaque rupture with intraluminal thrombus formation (65.21% vs. 39.13%, P = 0.1392). In vivo experiments indicated that NR1D1 exerted a protective role in the vasculature. Mechanically, NR1D1 deficiency aggravates macrophage infiltration, inflammation, and oxidative stress. Compared with the ApoE-/- mice, NR1D1-/-ApoE-/- mice exhibited a significantly higher expression level of pyroptosis-related genes in macrophages within the plaque. Further investigation based on mice bone marrow-derived macrophages (BMDMs) confirmed that NR1D1 exerted a protective effect by inhibiting macrophage pyroptosis in a NLRP3-inflammasome-dependent manner. Besides, pharmacological activation of NR1D1 by SR9009, a specific NR1D1 agonist, prevented plaque vulnerability/rupture. In general, our findings provide further evidences that NR1D1 plays a protective role in the vasculature, regulates inflammation and oxidative stress, and stabilizes rupture-prone vulnerable plaques.


[Healthcare research in dermatology 2018 - power of the factual : What healthcare research is and who it serves].
In the dermatology practiced in Germany, healthcare research is an established and highly differentiated discipline. In many university dermatological institutions and private practices, studies are carried out on the processes of healthcare research for skin diseases. Commissioned by the German Society of Dermatology and the Professional Association of German Dermatologists, a significant number of the studies and projects are conducted by the German Center for Health Services Research in Dermatology (CVderm) as a nationwide reference center. Important projects include the implementation of patient registers and national care studies, research with secondary data and carrying out methodological studies for the further development of scientific questions. Important results are the national healthcare conferences for psoriasis, skin cancer, wounds and atopic dermatitis, the regional care networks as well as projects and cooperation with health insurance companies, local government and public institutions. In the interests of improved patient care, important factors of care are continuously improved. Future health services research in dermatology will be able to contribute even more efficiently for better care planning and management. Patient registers will facilitate the transfer of innovations into care and help to ensure that new treatment approaches can be implemented. Registers also support the optimization of therapy approaches, which can no longer be carried out on the basis of clinical studies alone, for example in the case of psoriasis, due to the large number of drugs available. In the era of digital medicine, healthcare research has an even more important function.


Comparison of the efficacy of two doses of dexmedetomidine as an adjunct to levobupivacaine in infraclavicular brachial plexus block: prospective double-blinded randomized controlled trial.
This prospective, double-blind, randomized, controlled trial compared the efficacy of two dexmedetomidine doses (50 and 100-μg) combined with levobupivacaine on sensory block duration in infraclavicular brachial plexus block. We hypothesized that perineural dexmedetomidine would extend sensory block duration dose-dependently. The study included 60 patients aged 20 to 60 years of both sex with an ASA I/II undergoing forearm and hand surgery. The patients were randomly assigned into three equal groups (n = 20) for ultrasound-guided infraclavicular brachial plexus block. The L group received 35-mL 0.5% levobupivacaine plus normal saline, the LD50 group received 35-mL 0.5% levobupivacaine plus 50-μg dexmedetomidine, and the LD100 group received 35-mL 0.5% levobupivacaine plus 100-μg dexmedetomidine. Patients were investigated for onset and duration of sensory blockade, time to first postoperative rescue analgesia, and the total 24-h postoperative morphine requirement. The LD100 group had a longer sensory block duration (15.55 ± 1.1 h; 95% confidence interval (CI), 15.04-16.06) than the LD50 group (12.8 ± 1.2 h; 95% CI, 12.24-13.36 h) (p < 0.001) or the L group (9.95 ± 1.05 h; 95% CI, 9.46-10.44 h) (p < 0.001). The LD100 group took longer to request postoperative rescue analgesia and required fewer postoperative morphine doses than the LD50 and L groups (P < 0.001). Sensory block duration was longer with perineural 100-μg dexmedetomidine as an adjunct to levobupivacaine than with 50-μg dexmedetomidine. This study was approved by the Ethics Committee of Aswan University Hospital (approval number: aswu/125/4/17) (date of registration: 04/04/2017). Furthermore, the trial was retrospectively registered at ClinicalTrial.gov (NCT04729868) with a verification date of January 2021.


Almorexant effects on CYP3A4 activity studied by its simultaneous and time-separated administration with simvastatin and atorvastatin.
To characterise further the previously observed cytochrome P450 3A4 (CYP3A4) interaction of the dual orexin receptor antagonist almorexant. Pharmacokinetic interactions were investigated (n = 14 healthy male subjects in two treatment groups) between almorexant at steady-state when administered either concomitantly or 2 h after administration of single doses of simvastatin (40 mg) or atorvastatin (40 mg). Almorexant dose-dependently increased simvastatin exposure (AUC0-∞) when administered concomitantly [geometric mean ratios (90 % CI): 2.5 (2.1, 2.9) (100 mg), 3.9 (3.3, 4.6) (200 mg)], but not Cmax [3.7 (3.0, 4.5) for both doses]. Time-separated administration resulted in relevant reductions of the interaction [AUC0-∞: 1.4 (1.2, 1.7) (100 mg), 1.7 (1.5, 2.0) (200 mg); Cmax: 1.5 (1.3, 1.9) (100 mg), 1.9 (1.6, 2.4) (200 mg)]. Similar results were obtained for hydroxyacid simvastatin. Independent of almorexant dose and relative time of administration, AUC0-∞ and Cmax of atorvastatin increased (ratios ranged from 1.1 to 1.5). AUC0-∞ and Cmax of o-hydroxy atorvastatin decreased dose-independently [AUC0-∞: 0.8 (0.8, 0.9) (100 mg), 0.6 (0.5, 0.6) (200 mg); Cmax: 0.3 (0.3, 0.4) (100 mg), 0.2 (0.2, 0.3) (200 mg)] when atorvastatin was concomitantly administered. Cmax of o-hydroxy atorvastatin slightly decreased (0.8 for both doses) following time-separated administration; AUC0-∞ was unchanged. Whereas almorexant increased simvastatin exposure dose- and relative time of administration-dependently, atorvastatin exposure increased to a smaller extent and irrespective of dose and time. This suggests that the observed interaction of almorexant with simvastatin is mainly caused by intestinal CYP3A4 inhibition, whereas the interaction with atorvastatin is more due to hepatic CYP3A4 inhibition.


A cephalometric assessment of children with fetal alcohol syndrome.
Lateral cephalometric radiographs were quantitatively assessed in a series of 15 American black children with fetal alcohol syndrome (FAS). Although none was profoundly affected, FAS had been diagnosed in all the children at birth. Comparisons with age-, sex-, and race-matched controls disclosed a triad of facial profile differences: (1) frontal bossing, (2) palatal plane tipped up in the front with proclined upper incisors and a sharp nasolabial angle acquired from digit habits, and (3) above-average length of the mandibular corpus. Collectively these generate the perception of midface hypoplasia, although the midface actually is unremarkable in size and position. A high prevalence of chronic digit habits (8 of 15) is a secondary consideration in FAS, leading to localized skeletodental problems.


Confirmation of clorsulon residues in cattle kidney by capillary gas chromatography-negative-ion chemical-ionization mass spectrometry.
A confirmatory assay for residues of the anthelmintic agent clorsulon [4-amino-6-(trichloroethenyl)-1,3-benzenedisulfonamide] in cattle kidney tissue has been developed. The assay involves isolation of a drug-containing fraction by solvent extraction, methylation of the analyte, and fused-silica capillary column gas chromatography-negative-ion chemical-ionization mass spectrometry of the pentamethyl derivative of clorsulon. The intensities of four negative ions [m/z 406 and 408 (trichloro species) and m/z 413 and 415 (dichloro species)] are monitored. Confirmation of the presence of drug in an analyte requires that all four ions appear at the appropriate retention time with their intensity ratios within 10-15% of those arising from analysis of the reference standard, methylated clorsulon; the lower limit of detection is 3 ppb. Quantification of the drug is based on the intensity of the m/z 406 ion. Identification and quantification of residues by the gas chromatographic-mass spectrometric assay gave results in good agreement with those obtained with an electron-capture gas chromatographic assay.


Controversies in diagnostic approaches to the indeterminate follicular thyroid nodule.
In this paper we review the controversies in diagnostic approaches to follicular thyroid nodules with indeterminate cytology. The authors consider some of the controversies surrounding the indeterminate follicular thyroid nodule, including the definition and the postoperative outcome. Among the techniques that may improve preoperative diagnostic accuracy are large needle aspiration biopsy and immunohistochemistry for galectin-3.


Homoharringtonine binds to and increases myosin-9 in myeloid leukaemia.
Homoharringtonine (HHT) is a natural alkaloid isolated from various Cephalotaxus species. HHT has been used to treat acute myeloid leukaemia (AML), chronic myeloid leukaemia (CML), chronic lymphocyte leukaemia and myelodysplastic syndromes. Although HHT inhibits protein synthesis and promotes apoptosis of leukaemia cells in preclinical studies, its molecular target proteins remain unknown. The aim of this study was to identify target proteins of HHT. We have synthesized a biotinylated affinity column and used it to identify targets of HHT and confirmed the results by MS and Western blots. We also examined the effects of HHT on the target protein and determined roles of the target protein in anti-leukaemia activities of HHT through Western blots, flow cytometry and retrovirus transfection. Myosin-9, a member of the myosin super-family, was identified as a direct interactor of HHT. Furthermore, HHT up-regulated the expression level of myosin-9 in both AML and CML cell lines in a time-dependent manner. Thus, HHT-induced apoptosis of leukaemia cells begins in 6 h and continues to increase for 24 h. There is a positive correlation between up-regulated myosin-9 expression level and increased percentage of apoptotic cells mediated by HHT. Overexpression of myosin-9 could increase the sensitivity of the leukaemia cells to the cytotoxicity of HHT and arrest cells in S and G2/M phases. Our results indicated that myosin-9 was the target protein of HHT and played an important role in the HHT-induced apoptosis of leukaemia cells.


Effect of abutment design on fracture resistance of resin-matrix ceramic crowns for dental implant restoration: an in vitro study.
The purpose of this study is to investigate the performance and fracture resistance of different resin-matrix ceramic materials for use in implant-supported single crowns with respect to the abutment design (crown thickness: 1 mm, 2 and 3 mm). Forty-eight abutments and crowns were fabricated on implants in the right lower first molar. Two resin-matrix ceramic materials for dental crowns were selected for study: (1) a glass-ceramic in a resin interpenetrating matrix (Vita Enamic, Vita, Germany) and (2) a resin-based composite with nanoparticle ceramic filler (Lava Ultimate, 3 M ESPE, USA). Three types of abutments were designed: 1 mm thick crown + custom titanium abutment, 2 mm thick crown + custom titanium abutment and 3 mm thick crown + prefabricated titanium abutment. The experiment was divided into 6 groups (n = 8) according to the crown materials and abutment designs. After 10,000 thermocycles, fracture resistance was measured using a universal testing machine. The statistical significance of differences between various groups were analysed with ANOVA followed by a post hoc Tukey's honestly significant difference test. The surfaces of the fractured specimens were examined with scanning electron microscopy (SEM). Two-way ANOVA revealed that the abutment design (F = 28.44, P = 1.52 × 10- 8<0.001) and the crown materials (F = 4.37, P = 0.043 < 0.05) had a significant effect on the fracture resistance of implant crown restoration. The Lava Ultimate-2 mm group showed the highest fracture resistance of 2222.74 ± 320.36 N, and the Vita Enamic-3 mm group showed the lowest fracture resistance of 1204.96 ± 130.50 N. Most of the 1 and 2 mm groups had partial crown fractures that could be repaired directly with resin, while the 3 mm group had longitudinal fracture of the crown, and the crowns were detached from the abutments. Based on the in vitro data of this study, the fracture resistance of the 2 mm thick resin-matrix ceramic crown design was higher than that of the 1 and 3 mm groups. The 2 mm thick resin-matrix ceramic crown and personalized abutment are an option to replace zirconia for implant crown restoration.


Implications of limiting mechanical thrombectomy to patients with emergent large vessel occlusion meeting top tier evidence criteria.
Recent guidelines for endovascular management of emergent large vessel occlusion (ELVO) award top tier evidence to the same selective criteria in recent trials. We aimed to understand how guideline adherence would have impacted treatment numbers and outcomes in a cohort of patients from a comprehensive stroke center. A retrospective observational study was conducted using consecutive emergent endovascular patients. Mechanical thrombectomy (MT) was performed with stent retrievers or large bore clot aspiration catheters. Procedural outcomes were compared between patients meeting, and those failing to meet, top tier evidence criteria. 126 patients receiving MT from January 2012 to June 2015 were included (age 31-89 years, National Institutes of Health Stroke Scale (NIHSS) score 2-38); 62 (49%) patients would have been excluded if top tier criteria were upheld: pretreatment NIHSS score <6 (10%), Alberta Stroke Program Early CT score <6 (6.5%), premorbid modified Rankin Scale (mRS) score ≥2 (27%), M2 occlusion (10%), posterior circulation (32%), symptom to groin puncture >360 min (58%). 26 (42%) subjects had more than one top tier exclusion. Symptomatic intracerebral hemorrhage (sICH) and systemic hemorrhage rates were similar between the groups. 3 month mortality was 45% in those lacking top tier evidence compared with 26% (p=0.044), and 3 month mRS score 0-2 was 33% versus 46%, respectively (NS). After adjusting for potential confounders, top tier treatment was not associated with neurological improvement during hospitalization (β -8.2; 95% CI -24.6 to -8.2; p=0.321), 3 month mortality (OR=0.38; 95% CI 0.08 to 1.41), or 3 month favorable mRS (OR=0.97; 95% CI 0.28 to 3.35). Our study showed that with strict adherence to top tier evidence criteria, half of patients may not be considered for MT. Our data indicate no increased risk of sICH and a potentially higher mortality that is largely due to treatment of patients with basilar occlusions and those treated at an extended time window. Despite this, good functional recovery is possible, and consideration of MT in patients not meeting top tier evidence criteria may be warranted.


Sociodemographic determinants of knowledge and attitude in the primary prevention of cervical cancer among University Tunku Abdul Rahman (UTAR) students in Malaysia: preliminary study of HPV vaccination.
Cervical cancer is the third most common cancer affecting women around the world in which the Human Papillomavirus (HPV) is the one of the recognized causative agent affecting women health. In response to this health issue, the Malaysian government had officially implemented the HPV immunisation programme for secondary schoolchildren in 2010 at the age of 13 years old and above. The purpose of this study is to investigate the sociodemographic determinants of knowledge and attitude among students of Universiti Tunku Abdul Rahman (UTAR) towards the HPV vaccination programme. A cross-sectional study was conducted using self-administered questionnaires, recruiting 374 UTAR's students as the respondents by using convenience sampling method. Respondents were categorized as having good/poor level of knowledge and positive/negative attitude towards HPV vaccination. Over half of the respondents were females (64.5%) and the majority were aged 20 years old and below (55.8%). Generally, 54.7% of the total respondents had a high level of knowledge towards HPV vaccine while 57.5% of the total respondents showed a negative attitude towards HPV vaccine. Female respondents aged 20 years old and below showed good knowledge (56.4%) and a more positive attitude (55.8%) towards HPV vaccine. Students from the Faculty of Medicine and Health Sciences (FMHS) exhibited higher knowledge (67.3%) and positive attitude (62.4%) as compared to the Faculty of Accountancy and Management (FAM) which showed only 32.7% of knowledge and 37.6% of positive attitude towards the HPV vaccination. The majority of UTAR students possess good knowledge regarding HPV vaccination. Nonetheless, they demonstrated a negative attitude towards HPV vaccination, depicting the necessity to impart and further intensify the sense of health awareness among all students, especially among male students. The judicious use of social media apart from the conventional mass media should be an advantage as to enhance the practice of HPV vaccination among them and thereafter minimize the health and economic burdens of cervical cancer.


Synthesis and application of fluorescent teixobactin analogs.
Teixobactin is a promising new antibiotic that kills a spectrum of Gram-positive pathogens that are considered to be urgent threats by the CDC and the WHO. Better understanding of the novel mechanism of action of teixobactin may assist in developing new antibiotics and furthering our understanding of antibiotic resistance. This chapter describes the synthesis and application of fluorescent teixobactin analogs in fluorescence microscopy to study the mode of action of teixobactin. The first part of this chapter describes the synthesis and purification of fluorescent teixobactin analogs using two synthetic approaches. The second part of this chapter describes the application of the fluorescent teixobactin analogs to visualize their interactions with molecular targets in B. subtilis using fluorescence microscopy. The methods described herein provide synthetic access to chemical probes that may help further the understanding of antibiotic resistance.


Syndecan 4 contributes to osteoclast differentiation induced by RANKL through enhancing autophagy.
Periodontitis is a common chronic disease. Osteoclast differentiation contributes to alveolar bone resorption which is a distinct phenomenon during periodontitis. Syndecan 4 (SDC4), a member of the syndecan family, was found to be highly expressed during periodontitis. However, little is known about its role in periodontitis. Herein, we explored the role of SDC4 in osteoclast differentiation. An experimental periodontitis rat model was established by ligating the right first molar. The SDC4 expression in periodontium was detected by western blot and immunofluorescence. Our study demonstrated that SDC4 was highly expressed in the periodontium of periodontitis rats. It was positively transcriptionally regulated by NF-κB. SDC4 silencing abrogated osteoclast differentiation induced by RANKL, while SDC4 overexpression enhanced osteoclast differentiation. Moreover, SDC4 enhanced autophagy induced by RANKL. 3-MA, an autophagy inhibitor, was employed to explore whether SDC4 impacts osteoclast differentiation through activating autophagy. Treatment with 3-MA abolished osteoclast differentiation which was enhanced by SDC4, indicating that SDC4 promotes osteoclast differentiation through activating autophagy. This study reveals that SDC4 may contribute to osteoclast differentiation during periodontitis through activating autophagy. It sheds light on the important role of SDC4 in periodontitis.


Glibenclamide blocks the relaxant action of pinacidil and cromakalim in airway smooth muscle.
Concentration-relaxation curves for pinacidil and cromakalim were obtained in isolated guinea-pig tracheas contracted by histamine. Pinacidil produced complete relaxation with an EC50 value of 2.8 microM. The antidiabetic sulfonylurea, glibenclamide (0.1, 1, 10 microM), caused concentration-dependent rightward shifts (3- to 16-fold) of the concentration-relaxation curve for pinacidil without changing the maximal relaxant response. Cromakalim produced 85% relaxation with an EC50 value of 1.1 microM. Glibenclamide (0.1 microM) displaced the concentration-relaxation curve to the right and at higher concentrations (1-10 microM) caused nearly complete suppression of the maximal relaxant response to cromakalim. Glibenclamide not only prevented the effects of pinacidil and cromakalim but also produced a concentration-dependent and complete reversal of submaximal relaxations produced by these drugs. Glibenclamide was a selective antagonist of the relaxation of airway smooth muscle induced by pinacidil and cromakalim. Concentration-relaxation curves for theophylline, terbutaline and verapamil were unaffected by glibenclamide.


How Do the Outcomes of Radiation-Associated Pelvic and Sacral Bone Sarcomas Compare to Primary Osteosarcomas following Surgical Resection?
Radiation-associated sarcoma of the pelvis and/or sacrum (RASB) is a rare but challenging disease process associated with a poor prognosis. We hypothesized that patients with RASB would have worse surgical and oncologic outcomes than patients diagnosed with primary pelvic or sacral bone sarcomas. This was a retrospective, multi-institution, comparative analysis. We reviewed surgically treated patients from multiple tertiary care centers who were diagnosed with a localized RASB. We also identified a comparison group including all patients diagnosed with a primary localized pelvic or sacral osteosarcoma/spindle cell sarcoma of bone (POPS). There were 35 patients with localized RASB and 73 patients with POPS treated with surgical resection. Patients with RASB were older than those with POPS (57 years vs. 38 years, p < 0.001). Patients with RASB were less likely to receive chemotherapy (71% for RASB vs. 90% for POPS, p = 0.01). Seventeen percent of patients with RASB died in the perioperative period (within 90 days of surgery) as compared to 4% with POPS (p = 0.03). Five-year disease-specific survival (DSS) (31% vs. 54% p = 0.02) was worse for patients with RASB vs. POPS. There was no difference in 5-year local recurrence free survival (LRFS) or metastasis free survival (MFS). RASB and POPS present challenging disease processes with poor oncologic outcomes. Rates of perioperative mortality and 5-year DSS are worse for RASB when compared to POPS.


[Memento mori: what can be learned from early paintings and woodcuts about death and dying in former times?].
What was the impression on death and dying from people living in the 15th century? To answer this question written information is rare on this topic, as few people were able to read at that time, but paintings and early woodcuts may be helpful. Danses macabres (Totentänze) could be seen in Tallinn (formerly Reval), Lübeck, Bern and other places: parts of the original dance macabre still exist in Tallinn, but those in Bern and Lübeck are destroyed; copies however may give a decent impression of their former appearance. At all these dances macabre the death invites persons for a dance: the pope, the Kaiser, the king, the queen, various noblemen and citizens, even young women and small children; to dance with the death meant to die. The death does not dance with any old person. At the time of these dances macabre epidemics and famines were frequent causes of untimely early death.--A booklet Ars moriendi was published about 1470 and taught people how to behave at their hour of death; various devils appear at the deathbed haggling for the soul of the dying person. Thereafter an angel convinces him to trust in god and to resist those false promises of the devil.Nowadays dying is quite different. Usually persons die at very old age and are frequently demented, they die in hospitals, even in intensive care units and possibly without attendance of family members. They may have suffered for a long time and have spent years in nursing homes. Today dying may be just a release from very long suffering.


Effect of substance P on exocrine secretion by rat submandibular gland cells.
The effects of the undecapeptide, substance P(SP), on the secretion of mucin and proteolytic enzymes from dispersed cells of the rat submandibular gland were studied. The peptide, at a concentration of 1 X 10(-7) M, stimulated the release of 31.9 +/- 3.0% (mean +/- SEM) of intracellular mucin over 40 min, compared with 12.5 +/- 1.5% in untreated controls (p less than 0.01). This effect was duplicated by the homologous peptides, physalaemin, and eledoisin-related peptide. Substance P action was not affected by pre-incubation of cells with phentolamine or propranolol and was therefore independent of adrenergic stimulation. Furthermore, SP did not enhance the intracellular concentrations of cyclic AMP or cyclic GMP, confirming that cyclic nucleotides were not involved in its stimulus-secretion coupling mechanism. The isoproterenol-stimulated secretion of mucin from dispersed cells was reduced to 75.7% of the normal response (p less than 0.01) after a brief exposure to SP. This inhibitory effect was probably mediated by intracellular events rather than by direct effects on cell surface receptors. However, mucin release after treatment with SP followed by norepinephrine (NE) was 161% of that caused by NE alone (p less than 0.01) and may reflect an additive response to the independent stimulation of SP and NE receptors. Substance P and related peptides had no effect on arginine esterase secretion in the experimental model, although a response was elicited by alpha- and beta-adrenergic agonists. It is, therefore, proposed that serous cells of the granular convoluted tubule in the rat submandibular gland lack substance P receptors.


A mutation at the major phosphotyrosine in pp60v-src alters oncogenic potential.
Previously (M.A. Snyder, J.M. Bishop, W.W. Colby, and A.D. Levinson, 1983, Cell 32, 891-901) a mutant was constructed in v-src in which the major phosphotyrosine site, tyr-416, was converted to phenylalanine. This mutant has now been examined both for tumorigenicity and a number of in vitro parameters relating to the transformed state and to the known properties of pp60v-src, the product of v-src. Mouse cells transformed by this mutant gene, which are called RSV-SF1, are tumorigenic only if tested in immunodeficient mice, whereas cells transformed by the wild-type parent are tumorigenic in either syngeneic or immunodeficient animals. When examined in vitro, RSV-SF1-transformed cells are virtually indistinguishable from cells transformed by wild-type pp60v-src. These findings raise the possibility that the protein kinase activity of pp60v-src may not be fully responsible for tumorigenesis by v-src, and moreover suggest that evasion of the host immune response is a necessary step in tumorigenesis by v-src.