Identifying spatio-temporal seizure propagation patterns in epilepsy using Bayesian inference.
Focal drug resistant epilepsy is a neurological disorder characterized by seizures caused by abnormal activity originating in one or more regions together called as epileptogenic zone. Treatment for such patients involves surgical resection of affected regions. Epileptogenic zone is typically identified using stereotactic EEG recordings from the electrodes implanted into the patient's brain. Identifying the epileptogenic zone is a challenging problem due to the spatial sparsity of electrode implantation. We propose a probabilistic hierarchical model of seizure propagation patterns, based on a phenomenological model of seizure dynamics called Epileptor. Using Bayesian inference, the Epileptor model is optimized to build patient specific virtual models that best fit to the log power of intracranial recordings. First, accuracy of the model predictions and identifiability of the model are investigated using synthetic data. Then, model predictions are evaluated against a retrospective patient cohort of 25 patients with varying surgical outcomes. In the patients who are seizure free after surgery, model predictions showed good match with the clinical hypothesis. In patients where surgery failed to achieve seizure freedom model predictions showed a strong mismatch. Our results demonstrate that proposed probabilistic model could be a valuable tool to aid the clinicians in identifying the seizure focus.


Population genetic structure of the freshwater snail, Bulinus globosus, (Gastropoda: Planorbidae) from selected habitats of KwaZulu-Natal, South Africa.
The freshwater snail Bulinus globosus is an important intermediate host of Schistosoma haematobium, the causative agent of urinary schistosomiasis. This disease is of major health concern, especially in Africa where the majority of cases have been reported. In this study the inter- and intra-genetic diversity and population genetic structure of B. globosus from nine locations in the KwaZulu-Natal province of South Africa was studied using four polymorphic microsatellite loci (BgZ1-BgZ4). Moderate genetic diversity was detected within populations with a mean diversity (HE) of 0.49±0.09. The majority of populations significantly deviated from Hardy-Weinberg equilibrium (p<0.05), due to a deficit of heterozygotes. Such deviations may be due to founder events that were caused by bottlenecks that occurred as a result of frequent droughts and flooding that these snails' habitats are exposed to. Overall, the populations studied seem to be partially inbreeders/selfers with mean estimates of 0.24/0.38. A discernable genetic structure was elucidated among populations as evident by the mean pairwise FST of 0.58±0.13. There was no significant association between genetic and geographical distance among populations, an indication of limited gene flow. This increases the chances of populations losing alleles due to genetic drift. Populations in close proximity demonstrated high genetic differentiation (58.77% total variation) due to allelic differences between them. The sample populations fell into 12 clusters, however, the populations from uMkhanyakude and uThungulu exhibited no discernable genetic structure. Genetically, the Bhobhoyi site found within the uGu district was equidistant to the two main sampling regions.


A spectroelectrochemical study on single-oscillator model and optical constants of sulfonated polyaniline film.
The optical properties of sulfonated polyaniline (SPAN) thin film prepared by electrochemical method have been investigated. Polychromic behavior of SPAN thin film (transparent yellow-green-dark blue) was observed when the cyclic voltammograms were taken between -0.25 V and +1.90 V (vs. Ag/AgCl, sat.) during the growth of polyaniline film. In situ UV-vis spectra of the polymers-indium tin oxide (ITO) glass electrode were taken during the oxidation of the polymers at different applied potentials. The direct band gap values of SPAN thin film changed from 3.771 eV to 3.874 eV with the applied potentials. From in situ UV-vis spectra, the optical constants such as refractive index and dielectric constant of the SPAN thin film were determined. The important changes in absorption edge, refractive index and the dielectric constant were observed due to the applied potentials. The refractive index dispersion curves of the film obey the single-oscillator model and oscillator parameters changed with the applied potentials. The most significant result of the present work is in situ spectroelectrochemical method, which can be used to modify the optical band gaps and constants.


Genomewide Association Study of Simvastatin Pharmacokinetics.
We investigated genetic determinants of single-dose simvastatin pharmacokinetics in a prospective study of 170 subjects and a retrospective cohort of 59 healthy volunteers. In a microarray-based genomewide association study with the prospective data, the SLCO1B1 c.521T>C (p.Val174Ala, rs4149056) single nucleotide variation showed the strongest, genomewide significant association with the area under the plasma simvastatin acid concentration-time curve (AUC; P = 6.0 × 10-10 ). Meta-analysis with the retrospective cohort strengthened the association (P = 1.6 × 10-17 ). In a stepwise linear regression candidate gene analysis among all 229 participants, SLCO1B1 c.521T>C (P = 1.9 × 10-13 ) and CYP3A4 c.664T>C (p.Ser222Pro, rs55785340, CYP3A4*2, P = 0.023) were associated with increased simvastatin acid AUC. Moreover, the SLCO1B1 c.463C>A (p.Pro155Thr, rs11045819, P = 7.2 × 10-6 ) and c.1929A>C (p.Leu643Phe, rs34671512, P = 5.3 × 10-4 ) variants associated with decreased simvastatin acid AUC. Based on these results and the literature, we classified the volunteers into genotype-predicted OATP1B1 and CYP3A4 phenotype groups. Compared with the normal OATP1B1 function group, simvastatin acid AUC was 273% larger in the poor (90% confidence interval (CI), 137%, 488%; P = 3.1 × 10-6 ), 40% larger in the decreased (90% CI, 8%, 83%; P = 0.036), and 67% smaller in the highly increased function group (90% CI, 46%, 80%; P = 2.4 × 10-4 ). Intermediate CYP3A4 metabolizers (i.e., heterozygous carriers of either CYP3A4*2 or CYP3A4*22 (rs35599367)), had 87% (90% CI, 39%, 152%, P = 6.4 × 10-4 ) larger simvastatin acid AUC than normal metabolizers. These data suggest that in addition to no function SLCO1B1 variants, increased function SLCO1B1 variants and reduced function CYP3A4 variants may affect the pharmacokinetics, efficacy, and safety of simvastatin. Care is warranted if simvastatin is prescribed to patients carrying decreased function SLCO1B1 or CYP3A4 alleles.


Inhibition of crosstalk between Bcr-Abl and PKC signaling by PEITC, augments imatinib sensitivity in chronic myelogenous leukemia cells.
Chronic myelogenous leukemia (CML), a clonal hyperproliferation of immature blood cells accounts for 20% of adult leukemia cases. Reciprocal translocation of chromosomes 9 and 22, results into Bcr-Abl fusion and is responsible for expression of a tyrosine kinase protein p210(bcr/abl), which mediates several survival pathways and confer therapeutic resistance. Protein kinase C (PKC), a family of serine threonine kinases play an important role in the process of leukemogenesis. A crosstalk between Bcr-Abl and PKC signaling has been documented. Therefore, targeting p210(bcr/abl) and its associated signaling proteins using non-toxic natural means will be an effective strategy for antileukemic therapy. Aim of the present study is to investigate whether PEITC, a natural isothiocyanate in combination with imatinib mesylate (IM), a tyrosine kinase inhibitor could increase the therapeutic efficacy of IM by modulating the expression of p210(bcr/abl). Enhanced cytotoxic efficacy of IM by PEITC was further validated using another myelogenous leukemia cell line, KU812. It was observed that PEITC in combination with IM efficiently downregulated the expression of p210(bcr/abl) in chronic myelogenous leukemia cell lines (K-562). PEITC inhibited the expressions of PKCα, PKCβII and PKCζ (both phosphorylated and total form). Expression of Raf1 and ERK1/2, two important target proteins in PKC signaling cascade was diminished. The result indicated that PEITC ultimately reduced expression of Raf1 and ERK1/2 through Bcr-Abl and PKC inhibition. This result was further confirmed by UCN-01, a selective PKC inhibitor and IM; indicating an association between p210(bcr/abl) and PKC with Raf1 and ERK1/2. PEITC thus may have enormous potential in synergistic therapy of leukemia by enhancing drug efficacy.


Functions of transcription factors NF-κB and Nrf2 in the inhibition of LPS-stimulated cytokine release by the resin monomer HEMA.
Resin monomers like 2-hydroxyethyl methacrylate (HEMA) interfere with effects induced by stressors such as lipopolysaccharide (LPS) released from cariogenic microorganisms. In this study, mechanisms underlying monomer-induced inhibition of the LPS-stimulated secretion of inflammatory cytokines from immunocompetent cells were investigated. Secretion of pro-inflammatory cytokines TNF-α, IL-6 and the anti-inflammatory IL-10 from RAW264.7 mouse macrophages exposed to LPS and HEMA (0-6-8mM) was determined by ELISA. The formation of reactive oxygen (ROS) and nitrogen species (RNS) was determined by flow cytometry (FACS) after staining of cells with specific fluorescent dyes. Cell viability was analyzed by FACS, and protein expression was detected by Western blotting. Secretion of TNF-α, IL-6 and IL-10 from LPS-stimulated cells increased after a 24h exposure. A HEMA-induced decrease in cytokine secretion resulted from the inhibition of LPS-stimulated NF-κB activation. Nuclear translocation of NF-κB was inhibited possibly as a result of enhanced levels of hydrogen peroxide (H2O2) and nitric oxide (NO) in HEMA-exposed cells. Oxidative stress caused by HEMA-induced formation of H2O2 and LPS-stimulated peroxynitrite (ONOO) also enhanced nuclear expression of Nrf2 as the major regulator of redox homeostasis, as well as Nrf2-controlled stress protein HO-1 to inhibit NF-κB activity. HEMA inhibited the LPS-stimulated expression of NOS (nitric oxide synthase) to produce NO but counteracted the expression of Nox2, which forms superoxide anions that combine with NO to peroxynitrite. Resin monomers like HEMA inhibit LPS-stimulated NF-κB activation essential for cytokine release as a crucial response of immunocompetent cells of the dental pulp to invading cariogenic pathogens.


Mitigating ammonia nitrogen deficiency in dairy wastewaters for algae cultivation.
This study demonstrated that the limiting factor to algae growth on dairy wastewater was the ammonia nitrogen deficiency. Dairy wastewaters were mixed with a slaughterhouse wastewater that has much higher ammonia nitrogen content. The results showed the mixing wastewaters improved the nutrient profiles and biomass yield at low cost. Algae grown on mixed wastewaters contained high protein (55.98-66.91%) and oil content (19.10-20.81%) and can be exploited to produce animal feed and biofuel. Furthermore, algae grown on mixed wastewater significantly reduced nutrient contents remained in the wastewater after treatment. By mitigating limiting factor to algae growth on dairy wastewaters, the key issue of low biomass yield of algae grown on dairy wastewaters was resolved and the wastewater nutrient removal efficiency was significantly improved by this study.


Calcium-mediated hemagglutination by serum amyloid P component and the inhibition by specific glycosaminoglycans.
Human serum amyloid P component (SAP) was found to agglutinate erythrocytes in the presence of calcium ion. The hemagglutination was strongly inhibited by hyaluronic acid as well as by heparan sulfate and dermatan sulfate, but not by chondroitin 4-sulfate and keratan sulfate. A specific binding of SAP to hyaluronic acid, heparan sulfate, and dermatan sulfate was also confirmed by the fact that these glycosaminoglycans blocked the binding of SAP to agarose, a specific ligand of SAP.


Maize transposable elements contribute to long non-coding RNAs that are regulatory hubs for abiotic stress response.
Several studies have mined short-read RNA sequencing datasets to identify long non-coding RNAs (lncRNAs), and others have focused on the function of individual lncRNAs in abiotic stress response. However, our understanding of the complement, function and origin of lncRNAs - and especially transposon derived lncRNAs (TE-lncRNAs) - in response to abiotic stress is still in its infancy. We utilized a dataset of 127 RNA sequencing samples that included total RNA datasets and PacBio fl-cDNA data to discover lncRNAs in maize. Overall, we identified 23,309 candidate lncRNAs from polyA+ and total RNA samples, with a strong discovery bias within total RNA. The majority (65%) of the 23,309 lncRNAs had sequence similarity to transposable elements (TEs). Most had similarity to long-terminal-repeat retrotransposons from the Copia and Gypsy superfamilies, reflecting a high proportion of these elements in the genome. However, DNA transposons were enriched for lncRNAs relative to their genomic representation by ~ 2-fold. By assessing the fraction of lncRNAs that respond to abiotic stresses like heat, cold, salt and drought, we identified 1077 differentially expressed lncRNA transcripts, including 509 TE-lncRNAs. In general, the expression of these lncRNAs was significantly correlated with their nearest gene. By inferring co-expression networks across our large dataset, we found that 39 lncRNAs are as major hubs in co-expression networks that respond to abiotic stress, and 18 appear to be derived from TEs. Our results show that lncRNAs are enriched in total RNA samples, that most (65%) are derived from TEs, that at least 1077 are differentially expressed during abiotic stress, and that 39 are hubs in co-expression networks, including a small number that are evolutionary conserved. These results suggest that lncRNAs, including TE-lncRNAs, may play key regulatory roles in moderating abiotic responses.


Profiling Glycoproteins on Functionalized Reverse Phase Protein Array.
Reverse phase protein array (RPPA), a high-throughput, parallel immunoassay in a dot-blot format, is a powerful tool to quantitatively profile protein expression in multiple samples simultaneously using small amounts of material. Despite its success, analysis of post-translationally modified (PTM) proteins has been limited in RPPA assays, primarily due to relatively low availability of antibodies specific to proteins of PTMs, e.g., glycosylation. Moreover, the high matrix complexity, with tens of thousands of proteins in cell lysates or tissue extracts and the low abundance of proteins with PTMs, makes it extremely challenging to detect these proteins with PTMs. Therefore, there is an urgent need to fill this gap, which would greatly contribute to the analysis of a specific PTM by RPPA. In this chapter, we introduce a novel RPPA platform, termed polymer-based reverse phase glycoprotein array (polyGPA), to measure the variation of glycosylation patterns on a three-dimensionally functionalized RPPA. Without the need of specific antibody towards glycosylation, polyGPA represents a highly sensitive strategy to analyze protein glycosylation in multiple complex biological samples in parallel.


Identification and characterization of antioxidant peptides from Chinese dry-cured mutton ham.
Chinese mutton ham is a dry-cured meat product with a long ripening time. The aim of this study was to identify and characterize antioxidant peptides from Chinese mutton ham. Mutton ham peptides (MHPs) were purified by gel filtration, anion exchange and reversed-phase high-performance liquid chromatography steps. The 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) free radical scavenging capacity was used to guide the purification of MHPs. Three antioxidant peptides were identified by liquid chromatography with tandem mass spectrometry (LC-MS/MS) as Met-Trp-Thr-Asp (MWTD), Ala-Pro-Tyr-Met-Met (APYMM) and Phe-Trp-Ile-Ile-Glu (FWIIE), with molecular weights 551.61, 611.76, and 706.84 Da, respectively. Among them, APYMM exhibited the highest ABTS radical scavenging activity. The three peptides had the ability to inhibit lipid oxidation and Fenton's reagent-induced protein oxidation and DNA damage. After simulated gastrointestinal digestion, FWIIE and APYMM showed increased antioxidant activity, while MWTD showed decreased activity. Three novel peptides isolated from Chinese mutton ham had strong biological activity. Chinese mutton ham is potentially a functional food and an excellent source of natural antioxidants. © 2019 Society of Chemical Industry.


Treatment decision and estimation of growth of head and neck paragangliomas.
Head and neck paragangliomas are slow growing tumors where observation has become more widely accepted. Tumor growth rate as well as predictors of increased tumor growth were analyzed with the goal to identify factors to better predict disease progression and counsel patients. Multi-institutional retrospective cohort study from 2011 to 2020. 130 head and neck paragangliomas in 125 patients were analyzed. 38 were observed (30.4%), 16 radiated (12.8%), and 71 underwent surgery (56.8%). Surgical patients were significantly younger (p = 0.038) and with more genetically mediated paragangliomas (p = 0.026). Significantly more patients were asymptomatic in the observation group (p = 0.005). Of the 39 observed tumors, 43.6% (n = 17) grew with a tumor doubling time of 5.67 years. More than half of the observed paragangliomas had no growth. When examining symptoms postoperatively and at follow-up, the surgical cohort had significantly more worsening symptoms (p = 0.007) and new cranial neuropathies (p = 0.031). Head and neck paragangliomas have slow growth rates if they grow at all. Patients in the surgical cohort had more clinical symptoms at presentation and worsening postoperative symptoms.


Relationship between myocardial bridges and reduced coronary atherosclerosis in patients with angina pectoris.
A myocardial bridge (MB) is a band of myocardium covering a coronary artery segment, typically located in the left anterior descending coronary artery (LAD). Bridged segments of the coronary artery are isolated from the influence of perivascular adipose tissue. The aims of this study were to investigate the relationship between MBs and atherosclerosis in bridged LAD segments and to evaluate whether perivascular adipose tissue is involved in this relationship. MBs were identified in the coronary arteries of patients referred for diagnostic cardiac CT. The calcium score of MBs of the LAD or, in patients without LAD-MBs, of a corresponding LAD segment at the same distance from its origin and over the same length was measured. Of 128 patients, 56 (44%) had in total 73 MBs. The mean MB length was 22 ± 14 mm and the median MB thickness was 0.8mm (interquartile range 0.3-2.1mm). MBs in the LAD were present in 40 patients (31%). The calcium score was 0 in 95% of the LAD segments with MBs compared with 52% of the corresponding LAD segments without MBs. The association between LAD-MBs and calcium score (OR 0.06, 95% CI 0.01-0.25) was not influenced by age and gender, but was attenuated by local perivascular adipose tissue thickness (OR 0.35, 95% CI 0.04-2.70). Coronary artery segments covered with an MB have a lower calcium score than segments without an MB. The association between MBs and calcium scores was influenced by local perivascular adipose tissue thickness.


Medical therapy of achalasia: A benefit reserved for few.
The rationale for medical therapy of initial achalasia and the results obtained over the last 20 years in our laboratory are presented. Achalasic patients were selected as candidates for medical therapy on the basis of the presence of a slight esophageal dilation (<5 cm) on X-rays and a good manometric response to nifedipine administration. These patients were asked to take 10-20 mg of nifedipine sublingually 30-45 min before each meal for 2 weeks. Chronic medical therapy was continued only in those with an 'excellent' or 'good' clinical response to nifedipine and a lack of severe side effects. X-ray controls were planned every 6 months and manometric examination after the first 6 months. Of the 56 patients selected in the above-mentioned manner, 17 had an insufficient clinical response or severe side effects during the initial trial and did not continue medical therapy. Of the 39 patients who started chronic medical treatment, 13 are still on therapy and 26 stopped after an average of 2.8 years: 17 because they underwent dilation or myotomy; 4 for unknown reasons, and 5 apparently recovered. Esophageal manometry was carried out in 4 of the latter patients and revealed that the achalasic motor pattern had been replaced by a near-normal pattern. We believe that medical treatment of achalasia should be carried out not only in those patients who cannot undergo invasive procedures or do not respond well to them, but also in patients with initial achalasia selected using the above-mentioned criteria, because regression of the disease could take place in some of them.


Multiresidue method for the simultaneous determination of veterinary medicinal products, feed additives and illegal dyes in eggs using liquid chromatography-tandem mass spectrometry.
A multiclass method was developed for the simultaneous determination of 120 analytes in fresh eggs. The method covers the analytes from the groups of tetracyclines (6), fluoroquinolones (11), sulphonamides (17), nitroimidazoles (9), amphenicols (2), cephalosporins (7), penicillins (8), macrolides (8), benzimidazoles (20), coccidiostats (14), insecticides (3), dyes (12) and others (3). Samples were extracted using 0.1% formic acid in acetonitrile:water (8:2) with the addition of EDTA and cleaned using solid phase extraction with Hybrid SPE cartridges. The chromatographic separation was achieved on C8 column using mobile phase consisting of (A) methanol:acetonitrile (8:2) - (B) 0.1% formic acid in a gradient mode. Validation results according to the Commission Decision 2002/657/EC are as follows: linearity (r⩾0.99), recovery (75-108%), repeatability (CV 1.60-15.9%), reproducibility (CV 2.60-15%), decision limit (CCα 2.25-1156 μg/kg) and detection capability (CCβ 2.04-1316 μg/kg). The presented method was used for analysis of 150 real eggs samples taken from monitoring control program.


Structure of the genome of Moloney murine leukemia virus: a terminally redundant sequence.
The genome of the Moloney strain of murine leukemia virus (Mo-MuLV) has been analyzed by digestion with ribonuclease T1 and separation of the digestion products by two-dimensional gel electrophoresis. Thirty large oligonucleotides isolated from such a fingerprint have been characterized. One of these oligonucleotides (number 21) was found to be present in twice the molar yield of the rest. The 30 oligonucleotides were mapped on the genome by determining their yields in various size classes of 3' terminal fragments of Mo-MuLV RNA. The physical map obtained in this way suggested that oligonucletoide 21 was present very near the 3' end of the geome as well as in another location near or at the 5' end. The genome structure suggested by these results was confirmed by analyzing oligonucleotides in Mo-Mulv RNA complementary to strong stop DNA, which is shown to be a copy of the 5' terminal 134 nucleotides of the MoMuLV genome. Some of the oligonucleotides in the RNA protected from RNAase digestion by hybridization to this DNA, including oligonucleotide 21, were present near both the 3' and 5' ends. Comparison of these with the nucleotide sequence of strong stop DNA shows that there is a terminal redundancy of 49-60 nucleotides in the Mo-MuLV genome RNA.


Diagnosis of subtle thoracolumbar burst fractures. A new radiographic sign.
A previously undescribed radiographic sign, the posterior vertebral body angle is successful in distinguishing subtle burst type fractures from compression fractures of the thoracolumbar spine. Correct diagnosis is imperative for proper treatment and accurate prognosis. When positive, this sign will alert the orthopaedist so that further diagnostic testing with computed tomography or sagittal tomograms may be performed. The sensitivity of this sign in diagnosing subtle burst type injuries is 75%.


Differences in the phospholipid, cholesterol, and fatty acyl composition of 3T3 and SV3T3 plasma membranes.
An analysis of the phospholipid, cholesterol, and phospholipid fatty acyl composition of isolated plasma membranes of 3T3 and SV3T3 mouse mebryo cells has been performed. The results show that the plasma membrane of SV3T3 cells contain relatively less phosphatidylethanolamine and sphingomyelin and more cholesterol than 3T3 plasma membranes. The fatty acyl composition of individual phospholipid classes as determined by gas liquid chromatography also showed differences between 3T3 and SV3T3 plasma membranes. The plasma membranes of SV40 transformed 3T3 cells contain: (a) a higher percentage of 18:1 and less 20:3 and 20:4 in phosphatidylethanolamine; (b) a higher percentage of 18:1 in phosphatidylserine; and (c) a higher percentage of 18:2 and 20:4 in phosphatidylinositol.


Predicting vaccine uptake during COVID-19 crisis: A motivational approach.
The present research examined which motivational factors contribute to individuals' intention to take a vaccine that protects against SARS-CoV-2-virus and their self-reported vaccine uptake several months later. The role of different types of motivation was investigated (i.e., autonomous and controlled regulation) as well as vaccine distrust and effort to obtain a vaccine. Across two large-scale cross-sectional (N = 8887) and longitudinal (N = 6996) studies and controlling for various covariates, autonomous motivation and distrust-based amotivation contributed positively and negatively, respectively, to a) concurrent vaccination intentions, b) self-reported vaccination and c) subsequent subscription to a waitlist to obtain a vaccine. Participants' infection-related risk perception predicted more positive vaccination outcomes through fostering greater autonomous motivation for vaccination and lower distrust, whereas pandemic-related health concerns failed to yield such adaptive effects. The results emphasize the importance of fostering autonomous motivation for vaccination and handling distrust.


Leydig cell hypoplasia: cases with new mutations, new polymorphisms and cases without mutations in the luteinizing hormone receptor gene.
Defective male sex differentiation in patients with hypoplasia of Leydig cells (LCH) is caused by deficient LH receptor signal transduction. To further investigate the variety of LH receptor gene mutations present in LCH patients and their influence on the phenotype, we examined 10 nonrelated patients with the clinical presentation of LCH. Ten patients with a clinical phenotype of LCH were analysed for mutations in the complete coding region of the LH receptor gene. Exons 1-10 and two overlapping fragments of exon 11 of the LH receptor gene including all intron-exon boundaries were amplified by polymerase chain reaction and sequenced. To screen for frequencies of DNA changes, mutation analysis was performed on 45-59 healthy persons using denaturation high-performance liquid chromatography. Six new DNA alterations were identified. Three of them appear to be new polymorphisms. A G to C change at the 28th nucleotide of intron 1 on one allele and a heterozygous CGA to CAA transition at codon 124 (R124Q) were found. Both findings in these two patients are polymorphisms that occur with a frequency of 17% and 1.7%, respectively. A silent heterozygous CTA to TTA change at codon 204 was identified. In a patient with micropenis, the analysis revealed a homozygous missense mutation at codon 625 (I625K). As reported previously, this alteration significantly impaired signal transduction and explains the partial phenotype. Finally, in one compound heterozygous patient, two different mutations were discovered. At the polymorphic site in exon 1, a 27-bp insertion (CTG)2 AAG (CTG)5 CAG and a premature stop codon in the transmembrane segment 4 (W491*) were found. Both mutations disrupt signal transduction and explain the complete phenotype of this patient. In five patients, no DNA alterations could be identified. Three mutations (33 bp insertion in exon 1; W491* and I625K) were identified that explain the phenotype in two patients. In addition, most of the patients with the clinical phenotype of LCH did not have causative mutations, suggesting that changes in other regions of the LH receptor gene, such as the large introns or the promoter region, may be responsible for the majority of cases. Alternatively, the displayed phenotype may be the result of other genetic defects. Our work further underscores the importance of thorough clinical analysis of patients before molecular analysis of a particular gene is performed.


[Osteoporosis in glucocorticoid-treated asthmatic patients. Part II. Dependence on asthma duration and its treatment as well as on type of applied steroids].
The purpose of this study was an attempt at determination whether the risk of osteoporosis in glucocorticoid-treated asthmatic patients depends on duration and type of steroid therapy. The examinations were performed in 82 patients aged 29-70 years, treated with different types of steroids for 1-25 years. In the analysed patients osteoporosis was established by measuring bone mineral density (BMD) of lumbar spine, assessed by quantitative computer tomography (QCT). Increased osteoporosis incidence and decrease of bone mass with the duration of bronchial asthma and corticoid therapy were demonstrated, and significantly higher osteoporosis incidence and severity were found in patients treated exclusively or predominantly with intramuscular steroid preparations.


Proceedings of the International Summit on Human Gene Editing: a global discussion-Washington, D.C., December 1-3, 2015.
The US Academies of Sciences and Medicine, the Royal Society, and the Chinese Academy of Sciences convened a summit of experts in biology, medicine, law, ethics, sociology, and journalism, in December 2015 to review the state of the art in gene editing technology and discuss the medical and social ramifications of the technologies. The summit concluded with the following consensus recommendations: (1) intensive basic and preclinical research in animal and human models should proceed with appropriate legal and ethical oversight; (2) clinical applications in somatic cells must be rigorously evaluated within existing and evolving regulatory frameworks for gene therapy; (3) it would be irresponsible to proceed with any clinical use of germline editing until relevant safety and efficacy issues have been resolved and there is broad societal consensus about such a use; and (4) the international community should strive to establish generally acceptable uses of human germline editing.


Structural elucidation of the cis-prenyltransferase NgBR/DHDDS complex reveals insights in regulation of protein glycosylation.
Cis-prenyltransferase (cis-PTase) catalyzes the rate-limiting step in the synthesis of glycosyl carrier lipids required for protein glycosylation in the lumen of endoplasmic reticulum. Here, we report the crystal structure of the human NgBR/DHDDS complex, which represents an atomic resolution structure for any heterodimeric cis-PTase. The crystal structure sheds light on how NgBR stabilizes DHDDS through dimerization, participates in the enzyme's active site through its C-terminal -RXG- motif, and how phospholipids markedly stimulate cis-PTase activity. Comparison of NgBR/DHDDS with homodimeric cis-PTase structures leads to a model where the elongating isoprene chain extends beyond the enzyme's active site tunnel, and an insert within the α3 helix helps to stabilize this energetically unfavorable state to enable long-chain synthesis to occur. These data provide unique insights into how heterodimeric cis-PTases have evolved from their ancestral, homodimeric forms to fulfill their function in long-chain polyprenol synthesis.


Drug abuse in South Carolina: comparison with three studies.
This report discusses results of a state study on drug abuse in South Carolina and compares them with those of a similar state study conducted the previous year, as well as with three national studies. Since the latter did not deal with all the specific aspects of the state study, only the relevant portions are compared. The first of the national studies is on assessment of the diffusion of heroin abuse to medium-sized American cities. The second is on teenage drug abusers admitted to treatment. The third is on students and drugs in colleges and high schools.


Zinc regulates primary ovarian tumor growth and metastasis through the epithelial to mesenchymal transition.
The trace element zinc plays an indispensable role in human health and diseases including cancer due to its antioxidant properties. While zinc supplements have been used for cancer prevention, zinc is also a risk factor for cancer development. It is still unclear how zinc plays a role in ovarian cancer. To understand how zinc contributes to ovarian tumor growth and metastasis, we examined whether zinc contributes to tumor metastasis by regulating epithelial to mesenchymal transition (EMT) using ovarian cancer cells in vitro. Cell migration and invasion were examined using transwell plates and EMT markers were examined using Western blot. Primary ovarian tumor growth and metastasis were assessed using orthotopic ovarian cancer mouse models in vivo. Zinc promoted EMT, while TPEN (N, N, N', N'-tetrakis-(2-pyridylmethyl)-ethylenediamine), a membrane-permeable selective zinc chelator, inhibited EMT in a dose dependent manner in ovarian cancer cells. Moreover, zinc promoted ovarian cancer cell migration and invasion, while TPEN inhibited cell migration and invasion. Zinc activated expression of the metal response transcriptional factor-1 (MTF-1), while TPEN inhibited MTF-1 expression in a dose dependent manner. Knockout of MTF-1 inhibited zinc-induced cell migration, invasion and augmented the inhibitory effect of TPEN on cell migration and invasion. Loss of MTF-1 attenuated zinc-induced ERK1/2 and AKT activation and augmented the effect of TPEN in attenuating the ERK1/2 and AKT pathways. TPEN effectively inhibited primary ovarian tumor growth and metastasis in an orthotopic ovarian cancer mouse model by suppressing EMT. zinc contributes to ovarian tumor metastasis by promoting EMT through a MTF-1 dependent pathway. Zinc depletion by TPEN may be a novel approach for ovarian cancer therapy by inhibiting EMT and attenuating the ERK1/2 and AKT pathways.


Correspondence factor analysis of steroid libraries.
The receptor binding of a library of 187 steroids to five steroid hormone receptors (estrogen, progestin, androgen, mineralocorticoid, and glucocorticoid) has been analyzed by correspondence factor analysis (CFA) in order to illustrate how the method could be used to derive structure-activity-relationships from much larger libraries. CFA is a cartographic multivariate technique that provides objective distribution maps of the data after reduction and filtering of redundant information and noise. The key to the analysis of very complex data tables is the formation of barycenters (steroids with one or more common structural fragments) that can be introduced into CFA analyses used as mathematical models. This is possible in CFA because the method uses X2-metrics and is based on the distributional equivalence of the rows and columns of the transformed data matrix. We have thus demonstrated, in purely objective statistical terms, the general conclusions on the specificity of various functional and other groups derived from prior analyses by expert intuition and reasoning. A finer analysis was made of a series of A-ring phenols showing the high degree of glucocorticoid receptor and progesterone receptor binding that can be generated by certain C-11-substitutions despite the presence of the phenolic A-ring characteristic of estrogen receptor-specific binding.


Assay of detergents by rocket electrophoresis in agarose gels containing red blood cells: "rocket hemolysis".
A method is described for quantitation of charged detergents using their hemolytic property in an electrophoresis assay in agarose gels containing red blood cells. After electrophoresis the zone of hemolysis is directly proportional to the concentration of detergent in the sample. Using this technique we have determined the smallest detectable concentration for the negatively charged detergents, sodium dodecyl sulfate (SDS) and Quil A to about 10 micrograms/ml and 25 micrograms/ml, respectively and the positively charged cetyltrimethylammonium bromide (CTAB) to about 10 micrograms/ml.


The effect of clipping frequency on the competitive interaction between two perennial grass species.
The effect of clipping frequency on competition between Lolium perenne and Agrostis tenuis was investigated. The yield of clippings of both species increased and then declined during the 12-week period of the experiment, but the clip yield of Lolium was always significantly greater than that of Agrostis. Lolium was clearly the better competitor in unclipped controls. The proportion of the biomass contributed to the mixture by Agrostis increased as the interval between clips decreased. Tiller production was unaffected by increased clipping frequency in Lolium but was increased in Agrostis. Total yield was much more drastically reduced by frequent clipping in Lolium than in Agrostis, where yield was practically unaffected by wide variations in clipping frequency.These results are in agreement with the field distributions of the two species. They also suggest that differences in height and response to clipping are likely to confound any attempt to monitor the progress of competition experiments by measuring the yield of clippings.


Spatial and temporal predictability of the parasite community structure of a benthic marine fish along its distributional range.
The search for consistent patterns of organisation in parasite communities remains a central theme in parasite community ecology. However, to date, much evidence comes from studies without replication in both space and time; when replicate communities are examined, repeatable patterns are rarely observed. Here we determine, using nested subset analyses, whether the infracommunities of ectoparasites and endoparasites of a benthic marine fish (Sebastes capensis) show non-random structure. Then we examine the spatial repeatability of parasite community structure across the host's distribution in the southern Pacific, and the temporal repeatability of ectoparasite community structure from one locality. In total, 537 fish were captured from different latitudes (between 11 degrees S and 52 degrees S) along the Pacific coast of South America; a further 122 specimens were captured in two other years from one of the sampling localities, Valdivia (40 degrees S). In spite of variation in fish sizes among samples, fish size generally did not correlate with either ecto- or endoparasite species richness. The ecto- and endoparasite species richness of the component communities were also not correlated with fish sample size across the nine localities. Significant nested patterns were found in the ectoparasite communities of S. capensis at all eight localities, except at latitude 52 degrees S. Significant nested patterns were also found in the endoparasite infracommunities of S. capensis at seven of the nine localities, the exceptions being those from latitudes 11 degrees S and 20 degrees S. On a temporal scale, significant nestedness was observed in the ectoparasite infracommunities of S. capensis during each of the 3 years of sampling at Valdivia. In general, the same parasite species are responsible for the repeatability of nested patterns, though their importance varies among localities. The spatial and temporal predictability of the parasite community structure in S. capensis may be associated with the fish's benthic habitat and territorial behavior, suggesting that host biology may be a key determinant of the structure of parasite communities.


In vivo evaluation of Adipose-Derived Stem Cells (ADSCs) using Nanofat technique and chitosan conduit for peripheral nerve defect repair.
Nerve autograft is the gold standard for reconstruction of peripheral nerve loss. The alternative is the interposition of a synthetic regeneration conduit. The purpose of the study was to evaluate the in vivo feasibility and interest of Adipose-Derived Stem Cells (ADSCs) using the Nanofat technique and chitosan conduit for peripheral nerve defect repair. In vivo, after the creation of a 10 mm defect of the sciatic nerve, 2 groups were defined according to the nerve repair in rats: "chitosan" group (n = 10) and "chitosan and ADSCs" group (n = 10) with a clinical and paraclinical evaluation at 7 weeks. The in vivo results seem to show that the adjunction of ADSCs was favorable clinically, histologically and functionally compare to a chitosan reconstruction alone. Peripheral nerve repair with defect using a chitosan conduit associated with ADSCs would constitute a surgical alternative in a single surgical step.


Improving radiologists' recommendations with computer-aided diagnosis for management of small nodules detected by CT.
To evaluate how computer-aided diagnosis (CAD) can improve radiologists' recommendations for management of possible early lung cancers on CT. Twenty-eight lung cancers and 28 benign lesions were employed. Each group of 28 lesions was classified into subgroups of two sizes (9 between 6 and 10 mm and 19 between 11 and 20 mm) and three patterns (8 with pure ground glass opacity [GGO], 12 with mixed GGO and 8 solid lesions). Sixteen radiologists participated in the observer study, first without and then with CAD. Radiologists' recommendations, including (1) follow-up in 12 months, (2) in 6 months, (3) in 3 months, or (4) biopsy, were compared at three levels of their malignancy probability ratings (low: 1%-33%; medium: 34%-66%; high: 67%-99%) for 896 observations (56 lesions by the 16 radiologists) in the two size subgroups and three patterns. The number of recommendations changed by radiologists by use of CAD was 163 (18%) among all 896 observations. Among these changed recommendations, the fraction showing a beneficial effect from CAD was 68% (111/163), and the fraction showing a beneficial effect regarding biopsy recommendations was 69% (48/70). With CAD, the radiologists' performance regarding biopsy recommendations was significantly improved for 43 lung cancers (31 changed to biopsy versus 12 changed away from biopsy; P = .003) and was also improved for 27 benign lesions (10 changed to biopsy versus 17 changed away from biopsy; P = .18). Most of the cancers with improved recommendations were solid lesions or mixed GGO and relatively large. CAD has the potential to improve the appropriateness of radiologists' recommendations for small malignant and benign lesions on CT scans.


Signs and symptoms associated with uterine cancer in pre- and postmenopausal women.
Uterine cancer is the most and second most common gynecological malignancy in developed and developing countries, respectively. The majority of endometrial cancers are diagnosed early due to the presence of abnormal uterine bleeding. The existing literature however contains only little data regarding the prevalence of such symptoms compared to patients with no or benign pathology. Therefore, a systematic review was conducted in order to determine the significance of various clinical signs and symptoms predicting uterine cancer. Embase, Web of Science and Medline databases were searched from inception until 18 June 2019. Studies eligible for selection inclusion assessed the diagnostic accuracy of clinical signs and symptoms in pre- and postmenopausal women aged 18-99 years old with uterine malignancy. Case reports, case series and studies of which full text was not available, were excluded. The risk of bias was assessed using the QUADAS-2 tool by two independent reviewers. Results were visualized by forest plots using RevMan(5.3). Forty-one studies were eventually included in this systematic review. Abnormal uterine bleeding occurring in pre-, post- and perimenopausal women was proven to be the most widely investigated symptom in relation to cancer of the uterus. Thirty-two articles examined patients with postmenopausal bleeding of which sensitivity and specificity varied between 0.28 to 0.86 and 0.63 to 0.84, respectively. Abnormal bleeding in pre- and perimenopausal women on the other hand showed a sensitivity ranging from 0.63 to 0.81. Its specificity could not be calculated due to missing data. Other symptoms appeared not sufficiently examined to assess their diagnostic accuracy range. This review highlights the current lack of knowledge regarding the diagnostic accuracy of several signs and symptoms for uterine cancer. After a thorough in-depth review of the literature, meta-analysis could not be performed due to the absence of control populations in the majority of articles. Further research is needed to establish the rule-in or rule-out value of specific clinical signs to identify patients at risk for uterine malignancy prompting further clinical assessment.


Cascade reaction biosensor based on Cu/N co-doped two-dimensional carbon-based nanozyme for the detection of lactose and β-galactosidase.
Developing high-performance nanozyme for the construction of high-sensitivity biosensor platform is of great significance. Herein, we have designed and synthesized a Cu/N co-doped carbon-based nanozyme by anchoring Cu atoms on two-dimensional carbon nanosheet (Cu/NC NS). The Cu/NC NS possessed ultra-high Cu loading, superior peroxidase-like catalytic activity, great stability and reusability. Significantly, the Cu/NC NS exhibited high affinity towards H2O2 and 3,3',5,5'-tetramethylbenzidine (TMB) under acidic condition, which could induce obvious color changes of TMB in the presence of low H2O2 concentration. By integrating the Cu/NC NS with β-galactosidase (β-Gal) and galactose oxidase (Gal Ox), a multienzyme cascade colorimetric sensing system was established for the detection of lactose and β-Gal, which realizing the assay of lactose and β-Gal in the linear ranges of 0.1-1.4 mM and 0.025-0.2 U/mL, respectively. And the limit of detection for lactose and β-Gal were 0.03 mM and 0.01 U/mL, respectively. Furthermore, this work achieved the accurate detection of the content of lactose in milk and the activity of β-Gal in human serum, exploiting a novel application for nanozyme in biosensing.


Biochemical and functional alterations associated with hypercholesterolemia in platelets from hypertensive patients.
Hypercholesterolemia and hypertension are two of the major risk factors associated with increased atherosclerotic vascular disease. An abnormal platelet function is one of the mechanisms proposed to participate in atherogenesis. This study was undertaken to find out whether hypercholesterolemia in hypertensive patients can change platelet lipid composition and reactivity. Twenty-nine untreated hypertensive patients were distributed into 3 age, body mass index and blood pressure-matched groups according to their plasma cholesterol levels (normal, borderline or elevated, group NC, BC and HC respectively). Their platelet lipid composition, cytosolic Ca2+ concentration, cyclic AMP content and aggregating response to ADP and collagen were determined. Platelet from group HC patients were characterized by reduced cyclic AMP content (evaluated in the presence and absence of a platelet phosphodiesterase inhibitor) and aggregating responses to ADP and collagen, increased palmitic acid content and decreased arachidonic, eicosapentaenoic and docosatetraenoic and pentaenoic acid content, resulting in a lowered polyunsaturated to saturated fatty acid ratio (P less than 0.001). In contrast, platelet cytosolic Ca2+ concentration, DPH steady-state anisotropy and cholesterol to phospholipid molar ratio were not significantly changed. This indicates that hypercholesterolemia is accompanied in hypertensive patients by marked changes in platelet fatty acid composition, cyclic AMP content and response to aggregating agents. These changes, which clearly differ from those induced by in vitro cholesterol loading, could reflect not only the balance between LDL and HDL stimulation but also an adaptation to hemodynamic perturbations.


Regulation of protein breakdown by epidermal growth factor in A431 cells.
Addition of epidermal growth factor (EGF) to cultures of A431 human epidermoid carcinoma cells produces an increase in the rate of intracellular protein breakdown that cannot be accounted for by increased proteolysis in lysates from EGF-treated cells. In support of this observation, inhibition of protein synthesis with cycloheximide does not reduce the EGF response in cell monolayers. On the other hand, inhibitors of lysosomal proteolytic function such as leupeptin, vinblastine and especially the weak base, ammonia, are able to block the ability of EGF to increase protein breakdown. Additional results suggest that the EGF effect is mediated via a stimulation of autophagy. First, the autophagocytosis inhibitor, 3-methyladenine, reduces the EGF response, and second, the ability of insulin to inhibit protein breakdown by preventing the formation of autophagic vacuoles is overcome by EGF. Moreover, the actions of inhibitors and competing hormones are similar to those reported for glucagon, a hormone known to increase autophagy. The EGF response on protein breakdown persists for at least 6 h after thorough washing of the A431 monolayers. This result contrasts with the rapid reversal of EGF effects in other cell lines. Examination of the fate of bound EGF in cells washed and incubated for 2 h at 37 degrees C shows that some 500-fold more EGF per mg protein is retained on the surface of A431 cells compared to AG2804-transformed fibroblasts, a difference which probably explains the unusual persistence of the EGF effect on protein breakdown.


Exercise training for competitive tennis.
This article reviews the muscle physiology underlying various forms of activity, as well as the adaptive responses induced by specific forms of training, and thus provides a rational basis for the design of a training program for competitive tennis.


Epidemiology and control of staphylococcal pyoderma among newborn infants: evaluation of a method for routine cord care with 4 per cent chlorhexidine-detergent solution.
The incidence and types of staphylococcal purulent lesions were prospectively studied among 3602 newborn infants. During an epidemic of post-discharge pyoderma, 87 per cent of all Staphylococcus aureus isolates from lesions could be traced to the nursery. The risk of post-discharge disease was related to the degree of staphylococcal skin colonization in the nursery. A method for routine cord care in the nursery with 4 per cent chlorhexidine-detergent solution was evaluated and proved to be effective in reducing staphylococcal colonization in the nursery as well as purulent skin disease at home. The clinical effectiveness of the regimen was followed up for 19 months and a persistent low infection rate was obtained among infants in the post-discharge period.


Validation of reflectance infrared spectroscopy as a quantitative method to measure percutaneous absorption in vivo.
Attenuated total-reflectance infrared (ATR-IR) spectroscopy has been used to follow the penetration of a model compound (4-cyanophenol; CP) across human stratum corneum (SC) in vivo, in man. CP was administered for periods of 1, 2, or 3 hr, either (a) as a 10% (w/v) solution in propylene glycol or (b) in an identical vehicle which also contained 5% (v/v) oleic (cis-9-octadecenoic) acid. At the end of the treatment periods, SC at the application site was progressively removed by adhesive tape-stripping. Prior to the removal of the first tape-strip, and after each subsequent tape-strip, an ATR-IR spectrum of the treated site was recorded. The presence of CP, as a function of position in the SC, was monitored spectroscopically via the intense C = N stretching absorbance at 2230 cm-1. The absolute amount of CP, as a function of SC depth, was determined by "spiking" the applied solutions with 14C-labeled compound and subsequent liquid scintillation counting of the removed tape-strips. The presence of oleic acid in the applied formulation significantly increased the rate and extent of CP delivery as evaluated by either spectroscopy or radiochemical analysis. Furthermore, the ATR-IR and direct 14C analysis of CP as a function of SC position were highly correlated. These data strongly support, therefore, the validation of ATR-IR as a quantitative tool to assess percutaneous penetration in vivo.


The noise absorption prediction of a combined and independent absorber under different conditions and at different frequencies, using the new Engineering Noise Control Software (ENC).
Noise is one of the most harmful factors in the work environment, which is very important to control. There are various techniques to achieve this goal. One of the most important of them is the use of noise absorbers. Absorbent materials are often used to counteract the effects of reflected noise from hard surfaces and reduce their level. This is an experimental-applied study conducted in the physical factor laboratory of the Faculty of Health. The purpose was to predict the Noise absorption rate of combined and independent absorbers under different conditions and frequencies using the new ENC (engineering noise control) software. The sound absorption determination was carried out in 5 stages, including sound frequency analysis for the source, measurement of the dominant frequency, measurement of the absorption coefficient of absorbent materials in different conditions, measurement of the limit frequency (peak frequency of noise absorption) and comparison of the software results with the findings of the impedance tube in Real conditions. The best absorption mode for combined and independent absorbents is using a 5 cm rock wool absorbent with a 2 cm thick air layer behind it without a polyurethane absorbent layer and a 10 cm wide rock wool absorbent with a 1 cm air layer behind it without polyurethane layer. A polyurethane layer on the stone wool absorber decreased the amount of noise absorption for high frequencies. The results obtained from the best absorption conditions in the ENC software were consistent with the findings from the impedance tube device in real situations. The results of this study showed that suitable and optimal conditions of sound absorption could be achieved by using the ENC software, correct design, use of suitable absorbers, changes in the physical parameters of the absorber, and the use of a combined absorber.


Carboxylated acrylonitrile butadiene-natural rubber latex blends with methyl methacrylate grafted natural rubber latex: mechanical properties and morphology.
Commercially available carboxylated acrylonitrile butadiene latex (XNBR) was physically blended with natural rubber latex (NR) at varying blend ratios to investigate its effect on the mechanical properties and morphology. Methyl methacrylate grafted natural rubber latex (MG) was added to the latex blends as a third polymer to study whether it could enhance the mechanical properties of the latex blend films. It was found that the tensile strength of the blend films irrespective of composition decreased when the two latexes were blended as compared to the virgin latex films. The modulus 300 decreased while the elongation at break and tear strength of the blend films increased gradually as the ratio of NR increased in the blend films. It was found that the MG did not enhance the mechanical properties of the XNBR/NR blend films under the current experimental condition. AFM phase imaging analysis revealed enhanced polymer distribution and evidence of NR-MG-XNBR interactions.


Organ- and species-specific accumulation of metals in two land snail species (Gastropoda, Pulmonata).
In order to evaluate the usefulness of terrestrial gastropods as bioindicators there is a need for studies that simultaneously compare (1) concentrations of metals in reference and polluted plots, (2) species within the same polluted habitat, (3) metal accumulation patterns in different organs and (4) metal accumulation patterns in relation to soil physicochemical properties. This study aims to assess metal accumulation patterns in two land snail species. Instead of analyzing an organism as a whole, investigating the partitioning of metals in different organs can provide information on the actual toxicological relevant fractions. Therefore, concentrations of Ag, Cd, Cr, Cu, Ni and Zn were examined in five different organs of Cepaea nemoralis, as well as in the foot and the body of Succinea putris. Snails were sampled at four polluted dredged sediment disposal localities and three adjacent less polluted reference plots situated along waterways in Flanders, Belgium. Due to the small size and problematic dissection of S. putris only the concentrations in the foot of both species could be compared. For this reason only, C. nemoralis can be described as a better bioindicator species that allows a far more detailed analysis of organ metal accumulation. This study showed that organs other than the digestive gland may be involved in the immobilization and detoxification of metals. Furthermore, pH, soil fractionation (clay %, silt %, sand %) and organic matter, correlate with metal accumulation in organs. However, most often the soil metal concentrations did not correlate with the concentrations found in snail organs. Metal concentrations in organs of both species (1) differed among polluted plots but rarely between polluted and reference plots within a locality, (2) were organ-specific (digestive gland>foot>albumen gland=spermoviduct=ovotestis), (3) were species-specific and (4) depended on the metal type (high Cd and Cu concentrations were observed in the digestive gland and foot respectively). Our study emphasizes that background metal levels should be taken into account when using invertebrates as bioindicators of metal contamination and that bioindicators may show substantial differences in accumulation patterns even if they have a highly comparable ecology.


The feasibility of using an American National Board examination for the evaluation of Swiss candidates for licensure.
A test in Internal Medicine of the American National Board of Medical Examiners was given to Swiss medical students as a graduating examination. This study dealing with the feasibility of such an enterprise describes the technical procedures and investigates the problems of translation and the validity of foreign made items. Comparison of item analyses for the 369 Swiss candidates and for a sample of 370 American candidates reveals that most items are of close comparability in difficulty, discrimination and pattern of response to the distractors. In a cross-national comparison of student's responses to the items no systematic content characteristics can be found for items favouring one group or the other. Swiss experts, in judging the validity of the items for testing Swiss students, could indeed identify in advance some of the items that proved to be less valid and more difficult for their--but also the American--students. It is concluded that National Borad examination can be as valid and suited to examine Swiss candidates for licensure as it is to examine American candidates.


Reducing red color intensity of seafood wastewater in facultative pond.
Studies were carried out on the growth of Chromatium sp. on seafood wastewater (SFWW), which under facultative conditions and light exposure produced red pigment. The strain grew and utilized organic matter in both dark and light exposure conditions, but it produced red pigment when exposed to light. The growth was repressed by aerobic condition. The red color intensity was reduced by about 32.5+/-1.5 and 70.8+/-2.8% when kept under dark and static conditions, or aerobic and light exposure conditions, respectively. The COD of SFWW and the number of cells of Chromatium sp. were also rapidly reduced by about 78.6+/-2.7 and 92.0+/-1.0%, respectively, under aerobic and light exposure condition. KNO3 and FeCl3 also reduced red color intensity and maximum removal of organic matter and red color were 30 and 4 mg/l, respectively. Aerobic conditions increased the color removal efficiency with 30 mg/l KNO3 and 4 mg/l FeCl3 treatments up to 96.5+/-1 and 98.9+/-1%, respectively.


Identification and characterization of small molecule modulators of KChIP/Kv4 function.
Potassium channels and their associated subunits are important contributors to electrical excitability in many cell types. In this study, a yeast two-hybrid assay was used to identify inhibitors such as a diaryl-urea compound (CL-888) that binds to and modulates the formation of the Kv4/KChIP complex. CL-888 altered the apparent affinity of KChIP1 to Kv4.3-N in a Biacore assay, but did not dissociate the two proteins in size-exclusion chromatography experiments. Kv4.2/KChIP1 current amplitude and kinetics were altered with compound exposure, supporting the hypothesis of a compound-induced conformational change in the protein complex. Fluorescence spectroscopy of a unique tryptophan residue in KChIP1 was consistent with compound binding to the protein. Molecular modeling using the KChIP1 crystal structure indicates that compound binding may occur in a small tryptophan-containing binding pocket located on the hydrophilic side of the protein.


Modulation of 'A'-type K+ current by rodent and human forms of amyloid beta protein.
The Alzheimer's disease related peptide amyloid beta (Abeta) might have a physiological role in upregulating K channel currents in neurones. Earlier studies used the human form of Abeta1-40 on rat neurones. We sought to confirm our hypothesis by use of rat Abeta, which has no Alzheimer's association. In rat cerebellar granule neurones and HEK293 cells expressing Kv4.2 subunits, whole-cell patch clamp of K currents revealed that preincubation of cells with recombinant human or rat Abeta1-40 (10 nM for 24 h) significantly increased K channel current density. This was accompanied by increased mRNA levels for Kv4.2. These data indicate that rodent and human Abeta are effective in modulating K currents. The effectiveness of nonaggregating rat Abeta also strongly supports a physiological role for the peptide.


Training primary health care workers in mental health and its impact on diagnoses of common mental disorders in primary care of a developing country, Malawi: a cluster-randomized controlled trial.
Mental health problems are common in primary care, with prevalence rates of up to 40% reported in developing countries. The study aim was to evaluate the impact of a specially designed toolkit used to train primary health care (PHC) workers in mental health on the rates of diagnosed cases of common mental disorders, malaria and non-specific musculoskeletal pains in primary care in Malawi. Clinics with out-patient services in the designated district were randomly divided into control and intervention arms. Using a two-phase sampling process, Self-Reporting Questionnaire scores, data on diagnoses made by PHC workers and results of the Structured Clinical Interview for DSM-IV for depression were collected from 837 consecutively attending adult patients in the pre-intervention study and 2600 patients in the post-intervention study. The point prevalence rates for probable common mental disorder and depression were 28.8% and 19%, respectively. Rates for both anxiety and depression diagnoses by PHC workers at baseline were 0% in both arms. Following training, there were significant differences between the two arms in the rates of diagnosed cases of depression [9.2% v. 0.5%, odds ratio (OR) 32.1, 95% confidence interval (CI) 7.4-144.3, p ≤ 0.001], anxiety (1.2% v. 0%, p ≤ 0.001) and malaria (31% v. 40%, OR 0.62, 95% CI 0.43-0.89, p = 0.01). The intervention arm had more cases diagnosed with depression and anxiety while the control arm had more cases diagnosed with malaria. Training of PHC workers in mental health with an appropriate toolkit will contribute significantly to the quality of detection and management of patients seen in primary care in developing countries.


High-Yield, Case-Based, Interactive Workshop on Telehealth and Teleneurology With Pediatric Resident Physicians.
Increasing prevalence of neurologic disorders with an aging global population and limited availability of neurologists may lead to worse patient outcomes. As a result of the COVID-19 pandemic, telehealth services surged, and despite easing public health measures, the demand has remained. Telehealth technology has the potential to close the physical gaps in expanding the reach of care. This academic half-day workshop sought to provide a learning opportunity in response to these concerns. The workshop consisted of small- and large-group case discussions among pediatric resident physicians (PGY 1-PGY 3) moderated by two child neurology faculty physicians over Zoom. Participants received a learner document with prereading articles and questions for each case. PowerPoint presentations with video demonstrations were used to introduce the cases and guide discussions. Of the 25 attendees, 14 (56% response rate) answered a nonmandatory postsession survey. Eighty-six percent of the respondents were very or extremely satisfied with the content covered and were similarly satisfied with the effectiveness of content delivery. Seventy-nine percent of the respondents found the content helpful or very helpful in preparation for the board, and 93% anticipated applying the content covered occasionally or frequently in their clinical practice. Small-group discussions with video demonstrations are helpful in increasing proficiency with telehealth technology and in examining board-relevant cases on pediatric patients. There is strong interest in subsequent telehealth half-day workshops that incorporate teaching through group discussions on relevant patient case scenarios.


Chemical and ecotoxicological properties of three bio-oils from pyrolysis of biomasses.
In view of the potential use of pyrolysis-based technologies, it is crucial to understand the environmental hazards of pyrolysis-derived products, in particular bio-oils. Here, three bio-oils were produced from fast pyrolysis of pine wood and intermediate pyrolysis of corn stalk and poultry litter. They were fully characterized by chemical analysis and tested for their biodegradability and their ecotoxicity on the crustacean Daphnia magna and the green alga Raphidocelis subcapitata. These tests were chosen as required by the European REACH regulation. These three bio-oils were biodegradable, with 40-60% of biodegradation after 28 days, and had EC50 values above 100mgL(-1) for the crustacean and above 10mgL(-1) for the alga, showing low toxicity to the aquatic life. The toxic unit approach was applied to verify whether the observed toxicity could be predicted from the data available for the substances detected in the bio-oils. The predicted values largely underestimated the experimental values.


Effectiveness of botulinum toxin injection in the treatment of de novo OAB symptoms following midurethral sling surgery.
Intravesical onabotulinumtoxinA (Botox®) is effective for idiopathic overactive bladder (OAB) symptoms. Our primary objective was to compare the efficacy of onabotulinumtoxinA for women with de novo OAB after midurethral sling (MUS) surgery and women with idiopathic OAB. Women enrolled in this prospective study had idiopathic (n = 53) or de novo (n = 49) OAB symptoms after MUS, with at least one episode of urgency urine incontinence per day. OnabotulinumtoxinA (100 U) was administered in 20 intradetrusor injections. Postvoid residual volumes were checked at 2, 4 and 12 weeks. Participants completed a 3-day bladder diary and the King's Health Questionnaire (KHQ) before and 12 weeks after treatment. After 12 weeks, 22 patients (41.5 %) in the idiopathic OAB and 19 patients (38.8 %) in the de novo OAB groups were completely dry. OnabotulinumtoxinA injections had a significant benefit within both groups (p <0.001) to decrease both the daily numbers of voids (-2.39 and -2.0) and incontinence episodes (-1.38 and -1.44), with no significant difference between groups. We observed an increase of mean voided volume of >90 ml in both groups. Urinary retention was observed in four patients. We observed similar improvement in OAB symptoms after intravesical onabotulinumtoxinA injections within both groups. The rates of retention and requirement for catheterization even for women with a prior MUS were acceptable. These observational data provide evidence that onabotulinumtoxinA can effectively treat patients with OAB following stress urinary incontinence surgery.


A Stable Heterocyclic Amino(phosphanylidene-σ(4)-phosphorane) Germylene.
A new stable heterocyclic germylene, in which the divalent germanium atom lies between a nitrogen atom and a phosphanylidene phosphorane group, was synthesized. Experimental and theoretical studies revealed the peculiar effect of phosphanylidene phosphorane substituent, which is a stronger π-donor towards germanium than an amino group is. Because of the weak phosphorus-germanium π-bond, this new germylene compound shows an enhanced reactivity compared to classical N-heterocyclic germylenes.


Exogenously administered leptin leads to weight loss and increased physical activity in the marsupial Sminthopsis crassicaudata.
The adipose tissue derived cytokine leptin, modifies energy balance via effects on both food intake and energy expenditure. It is not clear, however, whether the component of energy expenditure accounted for by voluntary (nonexercise) physical activity is increased in response to leptin. The aim of this study was to investigate the effect of exogenously administered leptin on physical activity in the marsupial Sminthopsis crassicaudata. Body weight, tail width and food intake, were measured daily and physical activity was measured hourly in normal lean S. crassicaudata (n=8) with ad libitum access to standard laboratory diet. After 5 days baseline the animals were divided into two equal groups (n=4), and either human recombinant leptin (2.5 mg/kg) or placebo was administered twice daily intraperitoneally. Approximately 81% of the total daily activity during the baseline period occurred during the nocturnal phase. After 9 days of leptin administration, there were significant decreases in body weight (P<0.001) and fat content (P<0.01), which were not accompanied by a decrease in total energy intake. Overall daily physical activity increased (P=0.028); this effect was confined to the dark phase (P=0.033). We conclude that in lean S. crassicaudata the exogenous administration of human recombinant leptin results in a decrease in adiposity which occurs in the absence of a measurable effect on food intake and is associated with an increase in non-exercise physical activity at least over the duration of this study.


Pertussis toxoid structure: a collaboration and comparison of two-dimensional liquid chromatography-tandem mass spectrometry, ultraperformance liquid chromatography-mass spectrometryE, and capillary liquid chromatography-matrix-assisted laser desorption ionization-tandem mass spectrometry.
The overall structure of pertussis toxoid has been established by analysis of its tryptic digest using two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS), capillary liquid chromatography-matrix-assisted laser desorption ionization-tandem mass spectrometry (CapLC-MALDI-MS/MS), and ultraperformance liquid chromatography-mass spectrometry(E) (UPLC-MS(E)). In addition to oxidation and hydrolysis of amino acids losses of terminal peptides are observed. On-line UPLC-MS(E) generated a similar sequence coverage as the other two methods that involved off-line fraction collection. In light of recent favorable comparisons to data-dependent acquisition, UPLC-MS(E) should be the initial method of choice for analysis of a peptide mixture of moderate complexity.


Dietary muramidase improved growth performance, feed efficiency, breast meat yield, and welfare of turkeys from hatch to market.
Muramidase is an enzyme that hydrolyzes peptidoglycans of bacterial cell walls and improves performance of broilers in a dose-dependent manner. An experiment was conducted to evaluate muramidase supplementation, at a high or step-down dose, in turkeys from hatch to market. Male, B.U.T. 6 turkey poults were placed in 24 floor pens at 32 birds per pen. Poults were fed 1 of 3 diets from d 1 to 126 of age. There were 8 replicate pens per treatment. The treatments were a control (CTL) diet, the CTL plus muramidase at 45,000 LSU(F)/kg from phase 1 to 6 (BAL45), and the CTL plus muramidase at 45,000 LSU(F)/kg from phase 1 to 3 and decreased to 25,000 LSU(F)/kg from phase 4 to 6 (BAL45-25). Data were analyzed using SAS. The model included treatment and block and means were separated by Fisher LSD test. Birds fed BAL45 were heavier (P < 0.05) and had a greater (P < 0.05) average daily gain compared with birds fed the CTL from hatch to d 126 of age. Birds fed BAL45-25 had a final BW and average daily gain intermediate to or equivalent to birds fed BAL45 at the same phases. Feed conversion ratio was improved (P < 0.05) in birds fed BAL45 compared with birds fed the CTL and intermediate in birds fed BAL45-25. Breast meat yield was greater (P < 0.05) in turkeys fed muramidase, regardless of dose, compared with birds fed the CTL. There was no effect of treatment on muramic acid content in the jejunum digesta or litter scores. The frequency of pododermatitis score 1 was greater (P < 0.05) and score 2 was lower (P < 0.05) in birds fed muramidase, regardless of dose, compared with birds fed the CTL diet. In conclusion, muramidase supplementation improved performance, breast meat yield, feed efficiency and some markers of welfare, proportional to the dose in the diets.


[Pigmented Paget's disease and pigmented breast cancer metastasis. Clinical and histologic simulation of malignant melanoma of the breast].
Pigmented epidermotropic breast carcinoma has to be included in the differential diagnosis of pigmented lesions of the breast. Two rare cases of pigmented Paget's disease and pigmented metastatic breast carcinoma are presented, which mimicked malignant melanoma in both clinical and histological criteria.


Hyaluronan-induced VEGF-C promotes fibrosis-induced lymphangiogenesis via Toll-like receptor 4-dependent signal pathway.
Hyaluronan (HA), a component of the extracellular matrix, modulates cellular behavior including angiogenesis. However, little is known about the effect of HA on lymphangiogenesis in fibrosis model. In this study, we investigated the roles of HA in lymphangiogenesis of unilateral ureteral obstruction (UUO). We found that HA cooperated synergistically with vascular endothelial cell growth factor-C to stimulate capillary-like tube formation and increase migration of cells in a haptotaxis assay. Accumulation of HA in the cortical interstitial space was positively correlated with the number of lymphatic vessels after UUO. Depletion of macrophages with clodronate decreased UUO-induced HA accumulation and lymphangiogenesis. Additionally, hyaluronan synthase (HAS) mRNA expression and HA production were increased in bone marrow-derived macrophages upon stimulation with TGF-β1. Transfer of mHAS2 and mHAS3 knock-down CD11b-positive macrophages to SCID mice resulted in a partial decrease in UUO-induced lymphangiogenesis. HA increased expression of vascular endothelial cell growth factor-C in macrophages. Vascular endothelial cell growth factor-C expression and LYVE-1-positive lymphatic area was significantly lower in the UUO-kidney from TLR4 null mice than that from TLR4 wild-type mice. Collectively, these results suggest that HA increases lymphangiogenesis in renal fibrosis model and also stimulates vascular endothelial cell growth factor-C production from macrophages through Toll-like receptor 4-dependent signal pathway.


Antihypertensive medication and their impact on cancer incidence: a mixed treatment comparison meta-analysis of randomized controlled trials.
The aim of this article is to determine the association between commonly used antihypertensive agents and the incidence of cancer. We conducted a mixed treatment comparison meta-analysis of randomized, controlled (placebo, active, or untreated control) trials of antihypertensive drugs. A systematic literature search was conducted through June 2007. The primary outcome measure assessed was the incidence of cancer. Mixed treatment comparison meta-analysis was used to combine direct, within-trial, and between-drug comparisons with indirect evidence from the other trials. The indirect comparisons, which safeguard within-trial randomized findings, were constructed from trials with one treatment in common. Results are reported as odds ratios with 95% credible intervals. In total, 27 studies (56 treatment arms, 126137 patients enrolled, and 5868 cancers identified during follow-up) were included in the base-case analysis. They were hypertension, prehypertension, heart failure, coronary artery disease, or renal disease trials and patients were randomized to one of the five major antihypertensive drug classes or to placebo/untreated control. With the placebo/untreated control group as the referent comparison, the odds for developing cancer were 0.99 (0.80-1.24) for angiotensin-converting enzyme inhibitors; 1.12 (0.87-1.47) for angiotensin receptor blockers; 1.00 (0.78-1.32) for beta-adrenergic blockers; 0.94 (0.73-1.19) for diuretics and 0.95 (0.79-1.13) for calcium channel blockers. In sensitivity analyses, the results were not altered to any noteworthy extent including exclusion of studies under 3 years duration. Based on the totality of the clinical trial literature, commonly used antihypertensive drugs are not associated with increased odds of developing cancer.


Asymptomatic Wolff-Parkinson-White Syndrome Should be Ablated.
Wolff-Parkinson-White syndrome (WPW) is associated with a small but lifetime risk of cardiac arrest and/or sudden cardiac death (SCD). However, the exact risk is not well defined, particularly in asymptomatic persons. Over recent years the authors have collected and reported new follow-up data among a large number of asymptomatic WPW patients, particularly children, intensively followed. These data have significantly contributed to the knowledge and definition of the natural history of WPW from childhood to adulthood. The risk of SCD is higher in asymptomatic children than in adults, and early ablation can be offered only to selected subjects after electrophysiologic testing.


The incidence, prevalence, and survival rate of thromboangiitis obliterans in Korea: a retrospective population-based study.
Few studies used nationwide data to assess the age-standardized prevalence rate, incidence rate, 10-year survival rate, and death risk of thromboangiitis obliterans (TAO; Buerger's disease). Data for 24,392 patients who had newly diagnoses related to TAO (I73.1) from 2006 through 2017 were extracted from the National Health Insurance Service in Korea. The age-standardized prevalence rate, incidence rate, 10-year survival rate, and death risk of TAO were analyzed. The mean (standard deviation) age of TAO patients overall was 62.0 (15.7) years; 61.3 (15.4) in males and 63.2 (16.1) in females (P<0.001). The proportion of patients older than 50 years old was about 80% overall. The proportion patients who died of TAO was 28.1%. Overall, the age-standardized prevalence rate of TAO decreased from 11.1 persons per 100,000 persons in 2006 to 8.43 persons in 2017, and overall, the incidence rate of TAO decreased from 6.07 persons in 2006 to 3.38 persons in 2017. The age-standardized prevalence rate and incidence rate in males were higher than that in females. The 10-year survival rate of TAO was about 65% (60.7% in males and 72.5% in females). The adjusted hazard ratio increased significantly with older age, male sex, hypertension, diabetes mellitus, myocardial infarction, heart failure, ischemic stroke, hemorrhagic stroke, chronic kidney disease, and malignant neoplasm. The 10-year survival rate of TAO was about 65%. Even though the age-standardized prevalence rate and incidence rate of TAO is decreased during the study period, the adjusted hazard ratio showed significantly increases with age and with male sex after adjustment for comorbidities.


The microtubule signature in cardiac disease: etiology, disease stage, and age dependency.
Employing animal models to study heart failure (HF) has become indispensable to discover and test novel therapies, but their translatability remains challenging. Although cytoskeletal alterations are linked to HF, the tubulin signature of common experimental models has been incompletely defined. Here, we assessed the tubulin signature in a large set of human cardiac samples and myocardium of animal models with cardiac remodeling caused by pressure overload, myocardial infarction or a gene defect. We studied levels of total, acetylated, and detyrosinated α-tubulin and desmin in cardiac tissue from hypertrophic (HCM) and dilated cardiomyopathy (DCM) patients with an idiopathic (n = 7), ischemic (n = 7) or genetic origin (n = 59), and in a pressure-overload concentric hypertrophic pig model (n = 32), pigs with a myocardial infarction (n = 28), mature pigs (n = 6), and mice (n = 15) carrying the HCM-associated MYBPC32373insG mutation. In the human samples, detyrosinated α-tubulin was increased 4-fold in end-stage HCM and 14-fold in pediatric DCM patients. Acetylated α-tubulin was increased twofold in ischemic patients. Across different animal models, the tubulin signature remained mostly unaltered. Only mature pigs were characterized by a 0.5-fold decrease in levels of total, acetylated, and detyrosinated α-tubulin. Moreover, we showed increased desmin levels in biopsies from NYHA class II HCM patients (2.5-fold) and the pressure-overload pig model (0.2-0.3-fold). Together, our data suggest that desmin levels increase early on in concentric hypertrophy and that animal models only partially recapitulate the proliferated and modified tubulin signature observed clinically. Our data warrant careful consideration when studying maladaptive responses to changes in the tubulin content in animal models.


Protelomerase uses a topoisomerase IB/Y-recombinase type mechanism to generate DNA hairpin ends.
Protelomerases are enzymes responsible for the generation of closed hairpin ends in linear DNA. It is proposed that they use a breaking-and-rejoin type mechanism to affect DNA rearrangement on specific DNA sequences. In doing so, one strand turns around and becomes the complementary strand. Using the purified enzyme from the Escherichia coli phage N15 and the Klebsiella phage phiKO2 and synthetic oligonucleotide substrates, we directly demonstrate the location where the cutting/re-ligation occurs. We identified a pair of transient staggered cleavages six base-pairs apart centered around the axis of dyad symmetry of the target site. Two molecules of the protelomerase form a pair of protein-linked DNA intermediates at each 3' end of the cleaved openings leaving a 5'-OH. Then, in a process not yet clearly defined, the partners of the two initial openings are exchanged, and the transient breaks are resealed to generate hairpin ends. The formation of 3'-covalent DNA-protein intermediates is a hallmark of the topoisomerase IB type reaction, and we have thus shown experimentally that protelomerase is a member of the tyrosine-recombinase superfamily. In addition, by introducing single nicks in the substrates as perturbation, we found that the integrity of the nucleotide chain 4 bp away from the cutting site as well as this nucleotide's complementary location on the stem if the strands were to fold into a cruciform structure are required for activity, suggesting that these locations may be important substrate-protein contacts. We determined that N15 and phiKO2 protelomerases are monomers in solution and two molecules are needed to interact with the substrate to form two closed hairpin products. The target sites of protelomerases invariably consist of inverted repeats. Comparative studies using the related target sites of different protelomerases suggest that these proteins may require both sequence-specific and structure (possibly cruciform)-specific recognition for activity.


Genomewide histone acetylation microarrays.
Histone acetylation and methylation are important regulators of gene activity. Chromatin immunoprecipitation (ChIP or ChrIP) has made it possible to examine not only the state of histone acetylation at a gene but also that of histone methylation and may soon be extended to other histone modifications such as phosphorylation and ubiquitination. In principle such studies are possible as long as an antibody is available to the particular histone modification. Once a target gene is identified it is instructive to see the effect of mutating putative enzymes responsible for the modification to determine how a particular enzyme is responsible for altering chromatin of that gene. Although specific target genes have been studied that contain such modifications recent technical advances have made it possible to study histone modifications genomewide. This not only allows for alternate views of particular paradigms to be investigated, but also uncovers chromosomal patterns of histone modification that would be missed in analyzing individual genes. We describe here an approach to rapidly study histone modifications genomewide by combining chromatin immunoprecipitation and DNA microarrays.


A phase I clinical trial of intraperitoneal thiotepa for refractory ovarian cancer.
Treatment options for patients with ovarian cancer who have failed systemic and intraperitoneal (ip) cisplatin-based chemotherapy are limited. We conducted a phase I clinical study of ip thiotepa in patients with refractory ovarian cancer to determine the maximum tolerated dose (MTD). Ten patients were given 39 courses of thiotepa (median number of courses per patient, 3.5; range, 1-10+). All patients had received prior ip cisplatin; 7 also had received iv cisplatin, and 5 had three or more prior regimens. Thiotepa (30-80 mg/m2) was given ip in 2 liters normal saline every 4 weeks. The therapy was well tolerated. There was no vomiting, stomatitis, alopecia, or peritonitis. The dose-limiting toxicity was myelosuppression. With repeated doses, patients had a delayed marrow recovery and required a 1- to 2-week delay in treatment. Six patients had stable disease (duration 2-14+ months; median duration 5 months); 1 patient had a 50% decrease in CA-125 level, and 1 patient with no measurable disease remained clinically disease-free. In summary, ip thiotepa had clinical activity in heavily pretreated patients with refractory ovarian cancer with disease stabilization seen in 6 of 9 evaluable patients and a partial response seen in 1 patient. Myelosuppression was the only toxicity encountered. A dose of 60 mg/m2 ip is recommended for phase II studies.


Velocity of axonal transport of labeled protein is not dependent on local concentration.
The velocity of axonal transport of protein labeled with [3H]leucine was determined in rat sciatic nerve sensory axons after reversible cooling of the nerve, which provoked a local accumulation of transported material. The velocity of the wavefront of the labeled protein, as well as the slope of the wavefront, was not affected by the duration of cold-block prior to rewarming. In addition, the velocity and slope did not change as the wavefront moved distally from the site of block. In these axons, therefore, velocity of axonal transport is independent of the local concentration of rapidly transported protein.


Reliability of the axial vertebral rotation measurements of adolescent idiopathic scoliosis using the center of lamina method on ultrasound images: in vitro and in vivo study.
This study aimed to investigate the intra- and inter-observer reliability of the axial vertebral rotation (AVR) measurements of adolescent idiopathic scoliosis (AIS) using the center of lamina (COL) method on ultrasound transverse images. Three cadaver vertebrae were scanned with 42 AVR configurations by both ultrasound and radiograph. In this in vitro study, four observers measured the AVR using the COL method on ultrasound transverse images and three observers measured the AVR using the Stokes' method on radiographs. In the in vivo study, 13 AIS subjects were recruited. Eighteen spinal curvatures were identified and 48 vertebrae were selected for the AVR measurements. Two observers performed the AVR measurements on both the ultrasound images and radiographs. All measurements were performed twice with 1 week interval apart to reduce memory bias. The intraclass correlation coefficient (ICC), mean absolute differences (MAD), and standard deviation (SD) were used to analyze the intra- and inter-observer reliability of the AVR measurements. The Bland-Altman plot was used to analyze the 95 % limit of the differences between the two methods. The proposed COL method had high intra- and inter-observer reliability on both the in vitro and in vivo studies (ICCs > 0.91, MADs < 1.4°) and agreed well with the experimental setup (ICCs > 0.96, MADs < 2.3°). The COL method showed good agreement with the Stokes' method for the in vitro study (ICC 0.84-0.85, MAD 4.5°-5.0°), while poor agreement for the in vivo study (ICC 0.49-0.54, MAD 2.7°-3.5°). The pilot study indicated the proposed COL method was a simple and reliable method to evaluate the AVR on ultrasound images. Standardization of the posture during ultrasound scan and taking radiograph is important.


Myeloperoxidase is a potential molecular imaging and therapeutic target for the identification and stabilization of high-risk atherosclerotic plaque.
As the inflammatory enzyme myeloperoxidase (MPO) is abundant in ruptured human atherosclerotic plaques, we aimed to investigate the role of MPO as a potential diagnostic and therapeutic target for high-risk plaque. We employed the tandem stenosis model of atherosclerotic plaque instability in apolipoprotein E gene knockout (Apoe-/-) mice. To test the role of MPO, we used Mpo-/-Apoe-/- mice and the 2-thioxanthine MPO inhibitor AZM198. In vivo MPO activity was assessed by liquid chromatography-tandem mass spectrometry detection of 2-chloroethidium generation from hydroethidine and by bis-5HT-DTPA-Gd (MPO-Gd) molecular magnetic resonance imaging (MRI), while plaque phenotype was verified histologically. Myeloperoxidase activity was two-fold greater in plaque with unstable compared with stable phenotype. Genetic deletion of MPO significantly increased fibrous cap thickness, and decreased plaque fibrin and haemosiderin content in plaque with unstable phenotype. AZM198 inhibited MPO activity and it also increased fibrous cap thickness and decreased fibrin and haemosiderin in plaque with unstable phenotype, without affecting lesion monocytes and red blood cell markers or circulating leukocytes and lipids. MPO-Gd MRI demonstrated sustained enhancement of plaque with unstable phenotype on T1-weighted imaging that was two-fold greater than stable plaque and was significantly attenuated by both AZM198 treatment and deletion of the Mpo gene. Our data implicate MPO in atherosclerotic plaque instability and suggest that non-invasive imaging and pharmacological inhibition of plaque MPO activity hold promise for clinical translation in the management of high-risk coronary artery disease.


Whole grain benefit: synergistic effect of oat phenolic compounds and β-glucan on hyperlipidemia via gut microbiota in high-fat-diet mice.
Increasing evidence has confirmed that whole grain oats are effective in regulating hyperlipidemia. However, which specific ingredient is crucial remains unclear. This study focused on which whole grain components, oat phenolic compounds (OPC) or oat β-glucan (OBG), can regulate lipid metabolism and gut microbiota. The experiment unveiled that OPC and/or OBG not only reduced the body weight and fasting blood glucose (FBG) but also regulated serum and hepatic lipid levels in high-fat-diet (HFD) fed mice. There was no significant difference in the regulatory effects of OPC and OBG (p > 0.05). The combination of OPC and OBG (OPC + OBG) significantly decreased the body weight (p < 0.01) and reduced the blood glucose (p < 0.01) and lipid profile levels (p < 0.01). The real-time quantitative PCR (RT-qPCR) study revealed that OPC + OBG significantly altered mRNA expression related to lipid metabolism. Histopathological analysis showed that OPC + OBG improved liver lipid deposition as well as liver oxidative stress (p < 0.05). In addition, OPC + OBG combination regulated the gut microbiota community phenotype and increased probiotics. OPC + OBG significantly increased the abundance of Bacteroidetes and reduced the abundance of Firmicutes (p < 0.05) compared with the OPC and OBG fed mice. In conclusion, OPC + OBG has a synergistic effect in alleviating hyperlipidemia via lipid metabolism and gut microbiota composition. This finding also provided a potential justification for the advantages of whole grains in preventing hyperlipidemia.


Regulation of body temperature in postmenopausal women: interactions between bromocriptine and the endogenous opioid system.
The role exerted by the endogenous opioid system on thermoregulation has been studied in nine postmenopausal women before and after the chronic administration of the dopamine agonist bromocriptine (5 mg/day). These women randomly received an infusion of the opioid antagonist naloxone (1.6 mg/h for 4 h) or saline on two consecutive days, before and after 30 days of bromocriptine administration. Body temperature as evaluated by rectal temperature, did not vary during saline infusion performed both before and after 30 days of bromocriptine administration. In untreated women naloxone infusion significantly reduced body core temperature. The hypothermic response to naloxone was significantly greater following chronic bromocriptine administration. These results indicate that bromocriptine seems to increase the activity of the endogenous opioid system on the mechanisms which regulate body temperature in postmenopausal women.


Tamsulosin and tolterodine in the medical expulsive therapy for intramural ureteral stones with vesical irritability: a prospective randomized study.
We performed a randomized, prospective study to assess the possible role of combined tamsulosin and tolterodine therapy for the relief of vesical irritability and in facilitating the spontaneous expulsion of intramural ureteral stones. Patients were randomized to one of three treatment groups. Treatment group 1 patients received tamsulosin 0.4 mg/day, group 2 patients received tamsulosin 0.4 mg/day plus tolterodine 2 mg (twice a day), and group 3 patients received tolterodine 2 mg (twice a day). Subjects rated the urgency associated with each micturition using the Urinary Sensation Scale. Pain descriptions were recorded by the patients using the Visual Analog Scale. Stone expulsion was observed in 30 patients in group 1, 29 patients in group 2 and 18 patients in group 3. Kaplan-Meier curves were plotted to access the expulsion rate of each group over time. A significant difference was shown for the expulsion rate between the tolterodine group and the other two groups. (P = 0.003 by log rank test). Average time to expulsion for groups 1, 2 and 3 was 7.62 ± 2.42, 7.79 ± 2.11 and 10.57 ± 2.71 days, respectively (P = 0.000). In groups 1, 2 and 3, the mean number of pain episodes was 2.27 ± 0.91, 1.39 ± 1.34 and 1.38 ± 1.20, respectively (P = 0.023). Treatment with tamsulosin and tolterodine appears to be beneficial in intramural ureteral stone clearance, particularly in intramural ureter stone with symptoms of vesical irritability.


Low-dose oral vitamin K therapy for the management of asymptomatic patients with elevated international normalized ratios: a brief review.
Asymptomatic elevation of the international normalized ratio (INR) is a common problem associated with hemorrhage. Evidence from randomized controlled trials supports the use of low-dose oral vitamin K therapy as a treatment that promptly reduces the INR. Vitamin K given orally is more effective than subcutaneous vitamin K injection, and as effective as intravenous administration when INR values are compared 24 hours after administration. A 1.0-mg vitamin K dose is likely most appropriate for patients with INR values between 4.5 and 10. The fear of over-correction of the INR has limited the widespread use of vitamin K; however, our review suggests that this occurs infrequently when small doses are administered orally.


Determination of bile acids in pharmaceutical formulations using micellar electrokinetic chromatography.
A micellar electrokinetic chromatography method (MEKC) has been developed and validated for the determination of bile acids (BA) such as ursodeoxycholic acid (UDCA), dehydrocholic acid (DHCA) and deoxycholic acid (DCA) in pharmaceuticals for quality control purpose. The background electrolyte consisted of 20 mM borate-phosphate buffer containing 50 mM sodium dodecylsulfate (SDS), and acetonitrile as additive. UV detection was set at 185 nm. Selectivity, linearity, range, repeatability, intermediate precision and accuracy showed good results. Comparison of the values obtained by MEKC and HPLC methods were in close agreement.