Evaluation of cyclic luciferin as a substrate for luminescence measurements in in vitro and in vivo applications.
Bioluminescence imaging (BLI) is a powerful tool for cell tracking, monitoring of gene delivery and expression in small laboratory animals. An alternative luciferase (Luc) substrate cyclic luciferin (Cycluc) was recently advanced for BLI applications as providing a stronger, more stable signal at significantly lower doses than the classical substrate D-luciferin (D-Luc) increasing sensitivity of Luc detection 10 to 100 times. We evaluated benefits of using Cycluc in in vivo studies in mice injected with murine adenocarcinoma 4T1 cells expressing Luc, and in single-cell organisms, the oocytes of Xenopus laevis. No significant increase in the efficacy of detection of the luminescent signal was recorded in either of the systems. Kinetic studies demonstrated that Km for Cycluc was 10000 higher, whereas Vmax was 100 lower than that of D-Luc. Cycluc efficiently bound to the active center of luciferase, but its turnover was extremely low, leading to actual inhibition of bioluminescence. This compromises Cycluc as a substrate for measurement of the activity of the wild-type luciferases, still widely used as reporters for in vivo monitoring microorganisms and tumor cells. It may find better applications with the development of in vivo imaging based on the genetically engineered mutant luciferases with different substrate requirements.


Taxonomic reconsideration of a sequestrate fungus, Octaviania columellifera, with the proposal of a new genus, Heliogaster, and its phylogenetic relationships in the Boletales.
During taxonomic revision of genus Octaviania in Japan we examined herbarium and fresh specimens of O. columellifera and O. asterosperma sensu S. Yoshimi & Y. Doi with morphological and molecular techniques. These two species were identical in both macro- and micromorphological characters and were clearly different from the generally known O. asterosperma. The identity of the two species and their distinctness from O. asterosperma was further supported by both nuclear large subunit and ITS rDNA phylogeny. The molecular analyses also revealed that O. columellifera shares its lineage with the boletoid mushroom-forming Xerocomus chrysenteron complex and that it does not form a monophyletic clade with other Octaviania species. Our morphological reevaluation, including transmission electron microscopic observation of basidiospores, clarified the taxonomic boundary between O. columellifera and other Japanese Octaviania species. Accordingly we propose a new genus, Heliogaster, for O. columellifera with designation of the lectotype. We discuss phylogenetic relationships with Octaviania sensu stricto species and the closely related boletoid (pileate-stipitate) fungi, generic characters of Heliogaster and intraspecific phylogeny.


Apoptosis signal-regulating kinase 1 mediates denbinobin-induced apoptosis in human lung adenocarcinoma cells.
In the present study, we explore the role of apoptosis signal-regulating kinase 1 (ASK1) in denbinobin-induced apoptosis in human lung adenocarcinoma (A549) cells. Denbinobin-induced cell apoptosis was attenuated by an ASK1 dominant-negative mutant (ASK1DN), two antioxidants (N-acetyl-L-cysteine (NAC) and glutathione (GSH)), a c-Jun N-terminal kinase (JNK) inhibitor (SP600125), and an activator protein-1 (AP-1) inhibitor (curcumin). Treatment of A549 cells with denbinobin caused increases in ASK1 activity and reactive oxygen species (ROS) production, and these effects were inhibited by NAC and GSH. Stimulation of A549 cells with denbinobin caused JNK activation; this effect was markedly inhibited by NAC, GSH, and ASK1DN. Denbinobin induced c-Jun phosphorylation, the formation of an AP-1-specific DNA-protein complex, and Bim expression. Bim knockdown using a bim short interfering RNA strategy also reduced denbinobin-induced A549 cell apoptosis. The denbinobin-mediated increases in c-Jun phosphorylation and Bim expression were inhibited by NAC, GSH, SP600125, ASK1DN, JNK1DN, and JNK2DN. These results suggest that denbinobin might activate ASK1 through ROS production to cause JNK/AP-1 activation, which in turn induces Bim expression, and ultimately results in A549 cell apoptosis.


DC-SIGN modulates DC maturation and function in rat renal tubulointerstitial lesions.
The role of DC-SIGN in tubulointerstitial lesions (TILs) and the effect of anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb) were investigated in rat nephrotoxic nephritis (NTN). On Day 4, immature DC-SIGN+DCs infiltrated into renal tubulointerstitium and matured by Day 14, showing increased migratory capacity and ability to induce T cell proliferation. The distribution of DC-SIGN+ DC significantly correlated with crescent formation, TIL severity, and changes in renal function. RANTES and TNF-alpha mRNA were continuously up-regulated from Day 4, while IL-10 mRNA was down-regulated after a marked increase on Day 4. Expression of IFN-gamma and IL-4 mRNA increased on Day 14 due to DC maturation. PsL-EGFmAb suppressed DC maturation, migration and ability to activate T cells. It also down-regulated TNF-alpha and up-regulated IL-10, resulting in a Th1/Th2 bias. The number of crescents decreased and TILs and renal function improved. These results suggest that DC-SIGN mediates DC tubulointerstitial infiltration and is an important regulator of local immune reactions and TILs. PsL-EGFmAb inhibited DC migration, maturation and function by targeting DC-SIGN, and may therefore be a potential treatment for NTN.


A biomarker for the assessment of niacin nutriture as a potential preventive factor in carcinogenesis.
The study of protective cellular responses to DNA damage has led to the working hypothesis that optimal niacin nutriture is a preventive factor in cancer. Described here is the development of a biomarker for determining niacin status termed Niacin Number. The combination of this biomarker with diet and cancer epidemiology will allow evaluation of the possible role of this nutrient in cancer risk.


Ruthenium Complexes Bearing Axially Chiral Bipyridyls: The Mismatched Diastereomer Showed Red Circularly Polarized Phosphorescence.
RutheniumII complexes bearing three axially chiral bipyridyl ligands were synthesized as a new family of chiral complex dyes, and Δ-(S)- and Λ-(S)-diastereomers were obtained. The X-ray crystal structure analyses, spectroscopy, and DFT calculations suggested that all the bipyridyls maintained chirality in both the ground and excited states, and the Δ-(S)- and Λ-(S)-isomers are the matched (more relaxed) and mismatched (more constrained) pairs, respectively. The mismatched Λ-(S)-isomer exhibited red circularly polarized phosphorescence (CPP) both in solution and in the solid state. The solution state CPP is the most intense of ruthenium complexes, while the solid state CPP is the first example of them. It is supposed that, for the Λ-(S)-isomer, the six cumulative CH/π interactions suppress further distortion in the T1 state.


Isolation and mapping of 328 new cosmid markers on human chromosome 8: construction of a high-resolution cytogenetic map of chromosome 8 with 416 markers.
We have determined the chromosomal localizations of newly isolated cosmids by fluorescent in situ hybridization (FISH) on prometaphase R-banded chromosomes and have constructed a high-resolution cytogenetic map for human chromosome 8 with 416 cosmid markers, including 328 new markers and 88 reported previously. Of the 416 markers, 229 were mapped to the long arm of chromosome 8, 181 to the short arm, and 6 to the centromere. Although the clones were scattered throughout the chromosome, they were concentrated in R-positive bands. Since the estimated physical length of chromosome 8 is 135 Mb, the overall average distance between loci is 320 kb, but the average separation of loci on R-positive bands is nearly 130-200 kb. This cytogenetic map will serve as a resource for efforts to characterize chromosomal and molecular aberrations involved in cancers, to clone genes associated with hereditary diseases, and to construct a detailed physical map of large electrophoretic fragments and/or contiguous cosmids and yeast artificial chromosomes.


Comprehensive Analysis of the SBP Family in Blueberry and Their Regulatory Mechanism Controlling Chlorophyll Accumulation.
SQUAMOSA Promoter Binding Protein (SBP) family genes act as central players to regulate plant growth and development with functional redundancy and specificity. Addressing the diversity of the SBP family in crops is of great significance to precisely utilize them to improve agronomic traits. Blueberry is an important economic berry crop. However, the SBP family has not been described in blueberry. In the present study, twenty VcSBP genes were identified through data mining against blueberry transcriptome databases. These VcSBPs could be clustered into eight groups, and the gene structures and motif compositions are divergent among the groups and similar within each group. The VcSBPs were differentially expressed in various tissues. Intriguingly, 10 VcSBPs were highly expressed at green fruit stages and dramatically decreased at the onset of fruit ripening, implying that they are important regulators during early fruit development. Computational analysis showed that 10 VcSBPs were targeted by miR156, and four of them were further verified by degradome sequencing. Moreover, their functional diversity was studied in Arabidopsis. Noticeably, three VcSBPs significantly increased chlorophyll accumulation, and qRT-PCR analysis indicated that VcSBP13a in Arabidopsis enhanced the expression of chlorophyll biosynthetic genes such as AtDVR, AtPORA, AtPORB, AtPORC, and AtCAO. Finally, the targets of VcSBPs were computationally identified in blueberry, and the Y1H assay showed that VcSBP13a could physically bind to the promoter region of the chlorophyll-associated gene VcLHCB1. Our findings provided an overall framework for individually understanding the characteristics and functions of the SBP family in blueberry.


Volume Matters: Improved Outcomes for Patients Presenting to High-Volume Emergency Departments with Atrial Flutter and Fibrillation.
Clinical familiarity plays a role in health outcomes; the relationship between emergency department (ED) volume and outcomes for atrial fibrillation and flutter (AFF) are not clear. We compared ED presentation outcomes for AFF between high (HV) and low volume (LV) EDs in Alberta, Canada. 45,372 AFF presentations for patients aged ≥ 35 years from all 104 EDs in Alberta during 1999 to 2011 using administrative health databases formed a retrospective cohort. EDs were grouped by annual AFF volume: 11 high (>100 presentations) or 93 low (≤100 presentations). Outcomes included hospital admission rate, return to ED for AFF within 30 and 90 days, and death within 30 and 90 days. Analyses included statistical tests and mixed effects modeling. Mean age at ED presentation was 69.8 years (52% male). HV ED presentations were associated with lower admissions (adjusted odds ratio [aOR] = 0.68, 95% confidence interval [CI] 0.64, 0.72; p-value [p]<0.001), ED returns at 90 (aOR = 0.81, 95% CI 0.73, 0.90; p<0.001) days, and a higher likelihood of specialist visits at 30 (aOR = 1.81, 95% CI 1.68, 1.94; p<0.001) and 90 (aOR = 1.82, 95% CI 1.76, 2.03; p<0.001) days. For admitted patients, there were fewer returns to HV EDs at 30 (aOR = 0.37, 95% CI 0.15, 0.87; p = 0.02) and 90 (aOR = 0.48, 95% CI 0.26, 0.89; p = 0.02) days after hospital discharge. There was no difference in death between the two groups. AFF patients presenting to HV EDs experienced fewer admissions and AFF ED revisit and higher specialist referrals compared to LV EDs.


Synthesis and order-disorder transition in a novel metal formate framework of [(CH₃)₂NH₂]Na(0.5)Fe(0.5)(HCOO)₃].
We report the synthesis, crystal structure, thermal, dielectric, Raman, infrared, and magnetic properties of [(CH3)2NH2][Na(0.5)Fe(0.5)(HCOO)3] (DMNaFe), the first metal formate framework templated by organic cations, presenting an ABO3 perovskite architecture with NaO6 octahedra as building blocks of the framework. On the basis of Raman and IR data, assignment of the observed modes to respective vibrations of atoms is proposed. We have found that DMNaFe undergoes a structural phase transition at 167 K on cooling. According to the X-ray diffraction, the compound shows R3[combining macron] symmetry at 293 K and triclinic P1[combining macron] symmetry at 110 K. The DMA(+) cations are dynamically disordered in the high-temperature phase and the phase transition is associated with ordering of the DMA(+) cations and distortion of the metal formate framework. The dielectric studies reveal pronounced dielectric dispersion that can be attributed to slow dynamics of the DMA(+) cations. Based on the low-temperature magnetic studies, this compound is a weak ferromagnet with a critical temperature 8.5 K.


Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta.
We report the cloning and characterization in tobacco and Arabidopsis of a Vigna radiata L. (mung bean) promoter that controls the expression of VR-ACS1, an auxin-inducible ACC synthase gene. The VR-ACS1 promoter exhibits a very unusual behavior when studied in plants different from its original host, mung bean. GUS and luciferase in situ assays of transgenic plants containing VR-ACS1 promoter fusions show strong constitutive reporter gene expression throughout tobacco and Arabidopsis development. In vitro quantitative analyses show that transgenic plants harboring VR-ACS1 promoter-reporter constructs have on average 4-6 fold higher protein and activity levels of both reporter genes than plants transformed with comparable CaMV 35S promoter fusions. Similar transcript levels are present in VR-ACS1 and CaMV 35S promoter lines, suggesting that the high levels of gene product observed for the VR-ACS1 promoter are the combined result of transcriptional and translational activation. All tested deletion constructs retaining the core promoter region can drive strong constitutive promoter activity in transgenic plants. This is in contrast to mung bean, where expression of the native VR-ACS1 gene is almost undetectable in plants grown under normal conditions, but is rapidly and highly induced by a variety of stimuli. The constitutive behavior of the VR-ACS1 promoter in heterologous hosts is surprising, suggesting that the control mechanisms active in mung bean are impaired in tobacco and Arabidopsis. The 'aberrant' behavior of the VR-ACS1 promoter is further emphasized by its failure to respond to auxin and cycloheximide in heterologous hosts. VR-ACS1 promoter regulatory mechanisms seem to be different from all previously characterized auxin-inducible promoters.


The endocannabinoid anandamide is a substrate for the human polymorphic cytochrome P450 2D6.
Members of the cytochrome P450 (P450) family of drug-metabolizing enzymes are present in the human brain, and they may have important roles in the oxidation of endogenous substrates. The polymorphic CYP2D6 is one of the major brain P450 isoforms and has been implicated in neurodegeneration, psychosis, schizophrenia, and personality traits. The objective of this study was to determine whether the endocannabinoid arachidonoylethanolamide (anandamide) is a substrate for CYP2D6. Anandamide is the endogenous ligand to the cannabinoid receptor CB1, which is also activated by the main psychoactive component in marijuana. Signaling via the CB1 receptor alters sensory and motor function, cognition, and emotion. Recombinant CYP2D6 converted anandamide to 20-hydroxyeicosatetraenoic acid ethanolamide and 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid ethanolamides (EET-EAs) with low micromolar K(m) values. CYP2D6 further metabolized the epoxides of anandamide to form novel dioxygenated derivatives. Human brain microsomal and mitochondrial preparations metabolized anandamide to form hydroxylated and epoxygenated products, respectively. An inhibitory antibody against CYP2D6 significantly decreased the mitochondrial formation of the EET-EAs. To our knowledge, anandamide and its epoxides are the first eicosanoid-like molecules to be identified as CYP2D6 substrates. Our study suggests that anandamide may be a physiological substrate for brain mitochondrial CYP2D6, implicating this polymorphic enzyme as a potential component of the endocannabinoid system in the brain. This study also offers support to the hypothesis that neuropsychiatric phenotype differences among individuals with genetic variations in CYP2D6 could be ascribable to interactions of this enzyme with endogenous substrates.


'I bet you know more and are nicer too!': what children infer from others' accuracy.
Research has shown that preschoolers monitor others' prior accuracy and prefer to learn from individuals who have the best track record. We investigated the scope of preschoolers' attributions based on an individual's prior accuracy. Experiment 1 revealed that 5-year-olds (but not 4-year-olds) used an individual's prior accuracy at labelling to predict her knowledge of words and broader facts; they also showed a 'halo effect' predicting she would be more prosocial. Experiment 2 confirmed that, overall, 4-year-olds did not make explicit generalizations of knowledge. These findings suggest that an individual's prior accuracy influences older preschoolers' expectations of that individual's broader knowledge as well as their impressions of how she will behave in social interactions.


Adenoviral vector-based strategies against infectious disease and cancer.
Adenoviral vectors are widely employed against infectious diseases or cancers, as they can elicit specific antibody responses and T cell responses when they are armed with foreign genes as vaccine carriers, and induce apoptosis of the cancer cells when they are genetically modified for cancer therapy. In this review, we summarize the biological characteristics of adenovirus (Ad) and the latest development of Ad vector-based strategies for the prevention and control of emerging infectious diseases or cancers. Strategies to circumvent the pre-existing neutralizing antibodies which dampen the immunogenicity of Ad-based vaccines are also discussed.


Cognitive error as the most frequent contributory factor in cases of medical injury: a study on verdict's judgment among closed claims in Japan.
Cognitive errors in the course of clinical decision-making are prevalent in many cases of medical injury. We used information on verdict's judgment from closed claims files to determine the important cognitive factors associated with cases of medical injury. Data were collected from claims closed between 2001 to 2005 at district courts in Tokyo and Osaka, Japan. In each case, we recorded all the contributory cognitive, systemic, and patient-related factors judged in the verdicts to be causally related to the medical injury. We also analyzed the association between cognitive factors and cases involving paid compensation using a multivariable logistic regression model. Among 274 cases (mean age 49 years old; 45% women), there were 122 (45%) deaths and 67 (24%) major injuries (incomplete recovery within a year). In 103 cases (38%), the verdicts ordered hospitals to pay compensation (median; 8,000,000 Japanese Yen). An error in judgment (199/274, 73%) and failure of vigilance (177/274, 65%) were the most prevalent causative cognitive factors, and error in judgment was also significantly associated with paid compensation (odds ratio, 1.9; 95% confidence interval [CI], 1.0-3.4). Systemic causative factors including poor teamwork (11/274, 4%) and technology failure (5/274, 2%) were less common. The closed claims analysis based on verdict's judgment showed that cognitive errors were common in cases of medical injury, with an error in judgment being most prevalent and closely associated with compensation payment. Reduction of this type of error is required to produce safer healthcare.


Black hair follicular dysplasia in Large Münsterländer dogs: clinical, histological and ultrastructural features.
Four Large Münsterländer cross-bred dogs affected with black hair follicular dysplasia (BHFD) and one unaffected control littermate were observed, and skin was sampled weekly over the first 19 weeks of life. Affected dogs were born with silvery grey hair, a consequence of melanin clumping in the hair shafts. Hair bulb melanocytes were densely pigmented, and contained abundant stage IV melanosomes but adjacent matrix keratinocytes lacked melanosomes. Melanin clumping was not prominent in epidermal melanocytes in the haired skin but occurred in the foot pads. Follicular changes progressed from bulbar clumping, clumping in the isthmus/infundibulum and finally to dysplastic hair shafts. Alopecia developed progressively in pigmented areas. Silver-grey hair, melanin clumping, accumulation of stage IV melanosomes within melanocytes and insufficient melanin transfer to adjacent keratinocytes are also classic features of human Griscelli syndrome. The underlying cause in Griscelli syndrome is a defect of melanocytic intracellular transport proteins leading to inadequate and disorganized melanosome transfer to keratinocytes with resultant melanin clumping. In view of the correlation in the phenotype, histology and ultrastructure between both disorders, a defect in intracellular melanosome transport is postulated as the pathogenic mechanism in BHFD.


[Possible use of the growing liver biological set for hepatic recovery after toxic damage (an experimental study)].
The lack of acceptable pharmacological approaches for restoration of the injured liver is associated with complex of mechanisms involved in hepatic regeneration and with difficulty of the target selection. The aim of this research was to study the hepatoprotective function of the extract from both the growing and regenerating liver containing a natural set of factors crucial for the hepatic restoration. Extracts from both regenerating liver of rats after 70% hepatic resection and the growing liver of neonatal pigs were obtained using own original technique. The set of resultant extracts was named as the hepatic regeneration set (HRS). HRS fractionation was carried out using the Toyopearl HW-50S sorbent. The efficiency of HRS and its fractions was estimated using a model of the mouse liver thioacetamide injury and monitoring hepatic enzyme activity in blood serum. The activities of AST and ALT in intact animals were 50 U/l and 80 U/l, respectively; after thioacetamide administration they increased to 2059±212 U/l and 4280±440 E/l, respectively (p<0.05). Treatment of injured animals with HRS from the rat regenerating liver resulted in a significant decrease of transaminase activities to 924±148 U/l (AST; p<0.05) and 1633±308 U/l (ALT; p<0.05). A similar effect was observed after treatment with HRS from the neonatal pig liver: the AST decreased to 937±138 U/l (p<0.05), while ALT activity decreased to 1710±237 U/l (p<0.05). HRs fractionation resulted in identification two active fractions characterized by much higher (8-29) hepatotropic effect that that of the whole extract. These fractions contained peptide/protein components with the range of molecular mass of 3-60 kDa (fraction 1) and 3-25 kDa (fraction 2a). Fraction 1 also contained some polynucleotides in fraction 1. Subsequent studies of these fractions exceeding the hepatotropic effect of original HRS is clearly needed to identify their individual components by immunochromatography methods, ELISA, MRM mass spectrometry and quantitative PCR.


Tissue inhibitor of metalloproteinases-1 (TIMP-1) modulates neuronal death, axonal plasticity, and learning and memory.
The tissue inhibitor of metalloproteinases-1 (TIMP-1) belongs to a family of multifunctional proteins that inhibit matrix metalloproteinases (MMPs), but also regulate cell growth, proliferation, migration and apoptosis in non-nervous tissues. We had previously reported that kainate (KA)-mediated excitotoxic seizures induce the expression of TIMP-1 in resistant neurons and reactive astrocytes of the rat CNS, but the functional implications of these changes had not been elucidated. In the present work we used a targeted gene null mutation in mice to investigate in vivo the involvement of TIMP-1 in neuronal death and axonal sprouting following KA. We found no differences in seizure behaviour between the wild-type (WT) and the TIMP-1 knock-out (KO) mice, without any compensation by other TIMPs, at least at the mRNA level. However, the TIMP-1 KO mice were resistant to excitotoxicity and did not undergo the typical mossy fibre sprouting observed in WT mice. The lack of TIMP-1 paradoxically hampered the increase in the activity of MMPs observed in the seizing WT mice. In addition, we demonstrate that learning and memory are impaired in untreated KO mice. In conclusion, this study provides the first in vivo evidence for the implication of TIMP-1 in neuronal death and axonal sprouting in a pathological situation, but also suggests the involvement of TIMP-1 in the synaptic mechanisms underlying learning and memory in physiological conditions. More generally, these data support the idea that the control of proteolysis is instrumental for pathological and physiological processes in the brain.


Uncoiling of a Cook Resonance Metallic Ureteral Stent.
Introduction and Background: The Cook Resonance® metallic ureteral stent's unique configuration allows adequate urine drainage while providing improved resistance to external ureteral compression. We report a case in which a stent broke and subsequently uncoiled during stent removal under cystoscopy. Case Presentation: A 46-year-old woman with a history of cervical carcinoma treated with radiation therapy and chemotherapy and bilateral ureteral strictures presented for cystoscopic removal of bilateral Cook Resonance ureteral stents. During removal of the right ureteral stent, a snap was felt with subsequent uncoiling of the stent. Firm tension to the inner wire and outside curls allowed eventual complete removal of the stent without any retained fragments. Discussion: The all-metal Cook Resonance stent is an attractive alternative to polymeric stents because of reduced frequency of stent exchange and resistance to external compression. However, the metal stent possesses risks such as increased urothelial hyperplasia and stent embedment.


A study of district nurses' experiences of continuous ambulatory chemotherapy.
As a result of technical developments and policies that promote shorter hospital stays, patients are increasingly receiving high technology treatment in the community. The administration of ambulatory intravenous chemotherapy at home is an example of such treatment. Despite being generalist nurses, district nurses (DNs) are involved in what could be viewed as 'specialist' care - advising and supporting patients while they are receiving treatment. This article reports on a study of 20 DNs from one community trust and examines the sources of the knowledge and skills used in caring for these patients and the communication links with the regional cancer centre.


Peptide and glycopeptide dendrimer apple trees as enzyme models and for biomedical applications.
Solid phase peptide synthesis (SPPS) provides peptides with a dendritic topology when diamino acids are introduced in the sequences. Peptide dendrimers with one to three amino acids between branches can be prepared with up to 38 amino acids (MW ~ 5,000 Da). Larger peptide dendrimers (MW ~ 30,000) were obtained by a multivalent chloroacetyl cysteine (ClAc) ligation. Structural studies of peptide dendrimers by CD, FT-IR, NMR and molecular dynamics reveal molten globule states containing up to 50% of α-helix. Esterase and aldolase peptide dendrimers displaying dendritic effects and enzyme kinetics (k(cat)/k(uncat) ~ 10(5)) were designed or discovered by screening large combinatorial libraries. Strong ligands for Pseudomonas aeruginosa lectins LecA and LecB able to inhibit biofilm formation were obtained with glycopeptide dendrimers. Efficient ligands for cobalamin, cytotoxic colchicine conjugates and antimicrobial peptide dendrimers were also developed showing the versatility of dendritic peptides. Complementing the multivalency, the amino acid composition of the dendrimers strongly influenced the catalytic or biological activity obtained demonstrating the importance of the "apple tree" configuration for protein-like function in peptide dendrimers.


Oxidants, antioxidant nutrients and the athlete.
Strenuous physical exercise induces oxidative stress. There may be a number of sources of this oxidative stress, including mitochondrial superoxide production, ischaemia-reperfusion mechanisms and auto-oxidation of catecholamines. Severe or prolonged exercise can overwhelm antioxidant defences, which include vitamins E and C and thiol antioxidants, which are interlinked in an antioxidant network, as well as antioxidant enzymes. Evidence for oxidative stress and damage during exercise comes from direct measurement of free radicals, from measurement of damage to lipids and DNA, and from measurement of antioxidant redox status, especially glutathione. There is little evidence that antioxidant supplementation can improve performance, but a large body of work suggests that bolstering antioxidant defences may ameliorate exercise-induced damage, suggesting that the benefits of antioxidant intervention may be for the long term rather than the short term.


Pesticide sprayers' knowledge, attitude and practice of pesticide use on agricultural farms of Ethiopia.
Pesticide sprayers from five state-owned agricultural farms took part in a study that evaluated their knowledge, attitude and practice of pesticide use on farms. Demographic features of the sprayers showed that the majority of them had primary-level education. The duration of work as sprayers on the farms ranged from 1 month to approximately 10 years, with 59% of them having worked as sprayers for <5 years. Their knowledge about hazards from pesticides was indicated by the responses they gave to the standard questionnaire. Careful working was considered to be very important by 93% of them, while only 7% suggested the use of personal protective devices (PPD). On some of the farms, sprayers were provided with inappropriate/unfit and worn-out PPD; 18% of the sprayers had unfit goggles and 29% used worn-out gloves. To minimize risk from pesticide application, 63% suggested avoiding applications during windy and sunny weather, 32% suggested the provision and proper use of PPD, while only 3% of them felt medical check-ups and training were important, and 2% suggested risks from spraying were best controlled by leaving their job. The hygiene and sanitation practices of the sprayers require much improvement. An attitudinal change is needed, together with the provision of better facilities and infrastructure. We recommend that pesticide safety education be given to the sprayers. In addition, appropriate PPD should be used, with regular maintenance and timely replacement of worn-out parts.


Localized inflammatory pulmonary disease in subjects occupationally exposed to asbestos.
In reviewing pathology materials from patients occupationally exposed to asbestos, we identified eight patients with either localized nodules in their lung or unusual pathologic changes. The chest radiographs of six patients showed isolated parenchymal nodules thought to represent primary neoplasms. In three cases, pathologic examination of these nodules showed intraluminal fibrosis and inflammation of the distal airways, a pattern of change frequently referred to as "bronchiolitis obliterans organizing pneumonitis." In each instance, asbestos bodies were present in association with the fibroinflammatory tissue. In one case, the nodule showed a desquamative interstitial pneumonitis type pattern, and asbestos bodies were present admixed with the alveolar macrophages and occasionally within their cytoplasm. In one case, the nodule was composed of nonspecific inflammation and fibrosis with focal bronchiolitis obliterans and frequent asbestos bodies scattered throughout the area of inflammation and fibrosis, and in another case, necrotizing inflammation association with Aspergillus fungal organisms was identified. Granulomatous inflammation was the dominant pulmonary pathologic change in one patient, and the other patient's lung biopsy specimen showed a diffuse lymphocyte-plasma cell interstitial pneumonitis. The cases reported suggest that asbestos may cause localized lesions in the lung that clinically and radiographically are misinterpreted as cancer and that pathologically show inflammation and fibrosis of the distal airways. In addition, our observations suggest that asbestos may cause granulomatous inflammation, a desquamative interstitial type pneumonitis, and a lymphocytic interstitial pneumonitis type pattern. Our conclusions that asbestos may cause these pathologic changes are supported by case reports in the clinical and pathologic literature, clinicopathologic studies, and by experimental studies.


Tinea capitis: still an unsolved problem?
Tinea capitis is a fungal infection specifically involving the scalp and hair. It is the most common dermatophyte infection in children under 12 years of age, with a predominance in those of sub-Saharan African descent. Common signs include hair loss, scaling, erythema and impetigo-like plaques. Adults may also be affected, but to a lesser degree. The causative species are from the Microsporum and Trichophyton genera. Limited treatment options and diverse modes of transmission complicate the clinician's ability to address this disease adequately. Although dermatophytes are ubiquitous in our environment and tinea capitis is common, therapeutic options can be utilised to reduce morbidity.


Pregnancy outcomes in women with endometriosis.
The aim of this study was to investigate whether patients with endometriosis who achieved pregnancy have a higher risk for pregnancy complications and adverse perinatal outcomes as compared to patients without endometriosis. The study compared obstetrical and perinatal outcomes of women with and without endometriosis who delivered between 1988-2013 at the Soroka University Medical Center. The study population included 504 deliveries divided into the following groups: 1) women with endometriosis (N.=35); and 2) without endometriosis (N.=467). Endometriosis was diagnosed by laparoscopy or laparotomy. Multiple logistic regression models were used to control for confounders. No significant increase in obstetrical complications was noted in the endometriosis group, but significantly higher rates of cesarean sections. Perinatal outcomes were comparable between the groups. In our population, endometriosis was not found as a risk factor for obstetrical complications or adverse perinatal outcomes. Larger population based cohort studies are needed to clarify the relationship between endometriosis and adverse pregnancy outcomes.


Occult bilateral involvement in stage I epithelial ovarian cancer.
The aim of this study was to determine the incidence of occult bilateral involvement in Stage I epithelial ovarian cancer. We retrospectively reviewed the records of all patients (n = 135) with Stage I epithelial ovarian cancer treated at the University of Pennsylvania Cancer Center between 1985 and 1996. A study group of patients (n = 118) who underwent a bilateral salpingo-oophorectomy as part of their staging laparotomy was identified. Operative notes, pathology reports, and discharge summaries were reviewed to document stage, grade, histology, intraoperative impression, gross pathological description, and microscopic diagnosis. Clinically occult bilateral involvement was defined as pathologically documented ovarian involvement by tumor in an ovary that appeared normal to the surgeon during staging laparotomy. The study group consisted of mostly Stage IA (79/118, 67%), followed by Stage IC (35/118, 30%) and Stage IB (4/118, 3%), tumors. In 9/118 (7.6%) of these cases, bilateral disease was documented in the pathology report. In 3/118 (2.5%) of these cases, the bilateral involvement by tumor was clinically occult at the time of surgery. In only 1/118 (0.85%) of these cases did a clinically abnormal appearing ovary not contain tumor. Occult involvement by early ovarian cancer is uncommon at the time of staging laparotomy. In appropriately counseled patients strongly desiring to preserve fertility, a unilateral salpingo-oophorectomy may be considered as part of a staging procedure for apparent Stage IA epithelial ovarian cancer if the contralateral ovary appears normal to the surgeon. Consideration should be given to a wedge biopsy with intraoperative frozen section to help exclude occult disease in the normal appearing ovary.


Sexy ladies sexing ladies: women as consumers in strip clubs.
Recent shifts in the consumer base of the sex industry have involved greater female attendance in strip clubs. This article examines how strip clubs and dancers incorporate female patrons into a sexualized space traditionally designed for men by identifying three interactional processes: passing over, sidestaging, and tailoring. We suggest dancers pass over women because they perceive female patron behavior to include resistance to "buying the game" and spending patterns that diverge from male customers. Drawing on Goffman's dramaturgical analysis, we suggest the dynamic relationship between dancer and female patron involves what we term sidestaging, which refers to both dancers' disclosure and how the club's spatial organization inhibits the construction of women as customers through sharing gendered spaces, such as the bathroom. We argue that when a dancer tailors her lap dance for a female patron, she succeeds in acknowledging the female customer's sexual subjectivity and potential same-sex desires by providing an individualized avenue for exploring an erotic experience. Finally, we discuss data implications for understanding how same-sex desire and sexual identity operate in an environment that eroticizes the female form, and how the strip club becomes a potential space for engaging in same-sex eroticism that includes elements of play.


Immunochemical studies of human prolidase with monoclonal and polyclonal antibodies: absence of the subunit of prolidase in erythrocytes from a patient with prolidase deficiency.
Prolidase was highly purified from human liver and erythrocytes. NaDodSO4/acrylamide gel electrophoresis revealed that these preparations contained a major protein with MW = 56,000. The mass of prolidase was estimated on gel filtration to be MW = 97,000, for both enzyme preparations. A monoclonal antibody was raised against the liver enzyme and a specific antiserum against the erythrocyte enzyme. The monoclonal antibody (EP-2) recognized prolidase from erythrocytes and liver, in equal proportions. The antiserum also recognized the enzyme from erythrocytes and liver. Immunoprecipitation studies with these antibodies suggested only a single species of prolidase in erythrocytes and liver. Using an immobilized monoclonal antibody (EP-2) as an immunoadsorbent, prolidase was partially purified from crude extracts, and the protein of the partially purified enzyme was identified by immunoblotting using antiserum. A protein band with a MW = 56,000 was demonstrated specifically when crude extracts from the liver and erythrocytes were examined using NaDodSO4/acrylamide gel electrophoresis. The subunit protein was absent in erythrocytes from a patient with prolidase deficiency. We propose that the absence of the subunit is one cause of the prolidase deficiency.


High Rates of Central Venous Line Replacement or Revision in Children With Cancer at US Children's Hospitals.
Most children with cancer utilize a central venous line (CVL) for treatment. Complications often necessitate early replacement, revision, or addition (RRA), but the rate of these procedures is not known. This study sought to determine rates of RRA in pediatric oncology patients, and associated risk factors. Data queried from the Pediatric Health Information System including patients ≤18 years old with malignancy and CVL placement. Analysis included: first CVL placement of the calendar year and subsequent procedures for 6 months thereafter. A total of 6553 children met inclusion criteria (55.9% male, median age 6 years, interquartile range: 2 to 12). RRA within 6 months was required in 25.6% of patients, with 1.7% requiring 5 or more lines. Patients with Central Line-Associated Bloodstream Infection (CLABSI) were 2.78 times more likely to require RRA within 6 months of initial CVL placement, but accounted for only 16% of RRA patients. Factors associated with RRA were age below 1 year, CLABSI, hematologic malignancy, malnutrition, clotting disorder, deep vessel thromboembolism, and obesity. Patients with implantable ports as initial CVL (42%) were less likely to need RRA. Twenty-five percent require at least 1 RRA within 6 months, with associated morbidity and costs. Though strongly associated, most revisions were not related to CLABSI episodes.


Correlational patterns of cerebral glucose metabolism in never-medicated schizophrenics.
We studied 18 never-mediated schizophrenic patients and 22 normal control subjects with 18F-deoxyglucose (FDG) positron emission tomography. Patients and controls performed the continuous performance test during FDG uptake. Cortical and subcortical structures comprising two circuits selected on the basis of several theoretical models of schizophrenia were examined. The correlation of glucose metabolic rate (GMR) for each structure in each circuit with connected structures was calculated and tested for two-tailed significance. Schizophrenics showed significantly different patterns of intercorrelations for both circuits. The largest difference was in the correlation of GMR in the anterior thalamus with the frontal cortex, a key element in the thalamo-cortical-striatal circuit suggested to be abnormal in some models of schizophrenia. Correlations of the frontal lobe with other regions were also more positive in normal controls than schizophrenics; controls had three correlational paths from the frontal cortex (to temporal cortex, ventral anterior thalamus, and dorsal medial thalamus) with significantly more positive correlations than schizophrenics perhaps consistent with other findings of frontal cortical dysfunction in schizophrenia. Normal controls also had both more significant positive and more significant negative correlations between the occipital cortex and other brain areas than schizophrenics. Correlations between homologous areas in the right and left hemispheres were prominent in both groups.


Topological dispositions of lysine alpha 380 and lysine gamma 486 in the acetylcholine receptor from Torpedo californica.
The locations have been determined, with respect to the plasma membrane, of lysine alpha 380 and lysine gamma 486 in the alpha subunit and the gamma subunit, respectively, of the nicotinic acetylcholine receptor from Torpedo californica. Immunoadsorbents were constructed that recognize the carboxy terminus of the peptide GVKYIAE released by proteolytic digestion from positions 378-384 in the amino acid sequence of the alpha subunit of the acetylcholine receptor and the carboxy terminus of the peptide KYVP released by proteolytic digestion from positions 486-489 in the amino acid sequence of the gamma subunit. They were used to isolate these peptides from proteolytic digests of polypeptides from the acetylcholine receptor. Sealed vesicles containing the native acetylcholine receptor were labeled with pyridoxal phosphate and sodium [3H]-borohydride. Saponin was added to a portion of the vesicles prior to labeling to render them permeable to pyridoxal phosphate. The effect of saponin on the incorporation of pyridoxamine phosphate into lysine alpha 380 and lysine gamma 486 from the acetylcholine receptor in these vesicles was assessed with the immunoadsorbents. The peptides bound and released by the immunoadsorbents were positively identified and quantified by high-pressure liquid chromatography. Modification of lysine alpha 380 in the native acetylcholine receptor in sealed vesicles increased 5-fold in the presence of saponin, while modification of lysine gamma 486 was unaffected by the presence of saponin. The conclusions that follow from these results are that lysine alpha 380 is on the inside surface of a vesicle and lysine gamma 486 is on the outside surface.(ABSTRACT TRUNCATED AT 250 WORDS)


Locking and unlocking of quantum nonlocality without entanglement in local discrimination of quantum states.
The phenomenon of nonlocality without entanglement (NLWE) arises in discriminating multi-party quantum separable states. Recently, it has been found that the post-measurement information about the prepared subensemble can lock or unlock NLWE in minimum-error discrimination of non-orthogonal separable states. Thus It is natural to ask whether the availability of the post-measurement information can influence on the occurrence of NLWE even in other state-discrimination strategies. Here, we show that the post-measurement information can be used to lock as well as unlock the occurrence of NLWE in terms of optimal unambiguous discrimination. Our results can provide a useful application for hiding or sharing information based on non-orthogonal separable states.


Pigmented Paget disease--a diagnostic pitfall mimicking melanoma.
Pigmented mammary and extramammary Paget disease are rare entities in both males and females that mimic melanoma both clinically and histologically. Furthermore, Paget disease can be associated with increased number of benign melanocytes, thus resulting in additional diagnostic difficulty, especially when only hematoxylin and eosin-stained sections are examined and a limited immunohistochemical study is performed. Using standard hematoxylin and eosin-stained sections and routine immunohistochemical studies, we describe and characterize 7 cases of pigmented extramammary and mammary Paget disease. In all cases, malignant epithelial cells showed intracytoplasmic pigment, along with an immunohistochemical epithelial phenotype. In 2 of the cases, immunohistochemistry revealed numerous dendritic processes positive for melanocytic markers, thus resulting in an initial diagnosis of melanoma. Careful analysis confirmed that the immunolabeling corresponded to cytoplasmic labeling of melanocyte dendrites surrounding tumor cells. The correct diagnosis of pigmented Paget disease can be reached after close histologic examination and detailed evaluation of immunohistochemical studies. The latter are especially important in some extraordinary cases in which there may be an associated intraepithelial melanocytic hyperplasia.


[Therapy of Benign Essential Blepharospasm with Botulinum Toxin].
Blepharospasm is a focal dystonia and a rare disease of the brain that causes involuntary spasms of the muscles responsible for eyelid closure. The diagnosis is made clinically and in unclear cases on the basis of results of electrophysiological tests. Therapy of choice consists of local injections with botulinum toxin that have to be repeated on a regular basis.


Velopharyngeal dysfunction: recommendations for use of nomenclature.
A review of literature relating to velopharyngeal dysfunction reveals that there is no consensus regarding the precise definition of the terms "velopharyngeal incompetence," "velopharyngeal inadequacy," or "velopharyngeal insufficiency." Since this represents an obstacle to communication among members of multidisciplinary teams concerned with rehabilitation of velopharyngeal problems, current usage of these terms is reviewed and recommendations for clarification of the nomenclature are provided.


Dual Regulatory Role of Chromatin Remodeler ISW1 in Coordinating Cellulase and Secondary Metabolite Biosynthesis in Trichoderma reesei.
The saprophytic filamentous fungus Trichoderma reesei represents one of the most prolific cellulase producers isolated from nature. T. reesei also produces a typical yellow pigment identified as sorbicillinoids during cultivation. Here, we identified an evolutionarily conserved histone remodeling factor, ISW1, in T. reesei that simultaneously participates in regulating cellulase and the yellow pigment biosynthesis. Trisw1 deletion almost abolished vegetable growth, asexual spore formation, and cellulase gene expression. However, its absence significantly enhanced the production of the yellow pigment. The observed dual regulatory role of TrISW1 was dependent on its ATPase activity. We demonstrated that Trisw1 disruption elevated the transcription of ypr1 coding for the transcriptional activator of sor genes encoding the polyketide synthases catalyzing the biosynthesis of sorbicillinoids but compromised that of xyr1 encoding the key transcriptional activator of cellulase genes. Discrete T. reesei homologous ISW1 accessory factors were also found to exert differential effects on the expression of these two types of genes. Further analyses showed that TrISW1 was recruited to cellulase gene promoters, and its absence interfered with loss of histone H4 at the cbh1 and eg1 promoters upon cellulose induction. To the contrary, Trisw1 deletion facilitated loss of H4 at the sor locus. These data indicate that TrISW1 represents an important chromatin remodeler with a dual role in coordinating the cellulolytic response and biosynthesis of the major secondary metabolite in T. reesei. IMPORTANCE Microorganisms, including Trichoderma reesei, constantly face the challenge to outcompete other species to ensure efficient colonization in their natural habitat. They achieve this usually by adopting two alternative strategies by either maintaining fast growth on limited nutrient resources or producing a versatile array of secondary metabolites to fight against competitors. These two strategies, however, have to be subtly controlled to balance the assignment of and thus make the best use of cellular resources. Here, we identified a chromatin remodeling factor, TrISW1, with a dual role in coordinating the cellulolytic response and biosynthesis of the major secondary metabolite in T. reesei. The data also provide a novel insight into how T. reesei takes advantage of a chromatin remodeler to exquisitely balance two different adaptive strategies to ensure an efficient allocation of cellular resources to achieve efficient colonization in a specific environment.


Serum haptoglobin concentration in growing swine after intranasal challenge with Bordetella bronchiseptica and toxigenic Pasteurella multocida type D.
The acute phase reaction, in association with progressive atrophic rhinitis (AR), was monitored for 3 wk using serum haptoglobin (HPT) quantification in thirty-six, 15 kg swine after intranasal challenge with varying doses of Pasteurella multocida type D (toxigenic strain) and Bordetella bronchiseptica. The challenge doses were administered alone or in combination with pigs divided into 9 isolated treatment groups. Increasing doses of B. bronchiseptica were associated with lower serum HPT (P < 0.05), whereas increasing doses of P. multocida tended to increase serum HPT (0.05 < P < 0.10). Significant and positive correlation of mean HPT and AR score was found in these pigs; increased AR scores were associated with elevated mean HPT concentration (r = 0.41, P < 0.01). A significant interaction between P. multocida and B. bronchiseptica dose indicated that increasing the dose of B. bronchiseptica, for a fixed P. multocida dose, was associated with less AR (P < 0.05). The AR scores were greater in pigs given P. multocida, than B. bronchiseptica alone. These results indicate that a complex interaction between Pasteurella multocida and Bordetella bronchiseptica causes progressive atrophic rhinitis and alters serum HPT concentration in swine.


Adverse effects of luteinizing hormone on fertility: fact or fantasy.
High tonic serum concentrations of luteinizing hormone (LH) in the follicular phase, frequently witnessed in polycystic ovary syndrome, have been associated with decreased reproductive function. Impaired rates of fertilization, conception and miscarriage are obtained when LH levels are high before oocytes are collected, during ovulation induction or in women with regular cycles. Conversely, treatment that decreases LH concentrations, such as gonadotrophin-releasing hormone analogue or laparoscopic ovarian puncture, eases induction of ovulation and pregnancy and improves miscarriage rates. Tonic hypersecretion of LH appears to induce premature oocyte maturation, causing the problems with fertilization and miscarriage.


Influence of Afforestation on the Species Diversity of the Soil Seed Bank and Understory Vegetation in the Hill-Gullied Loess Plateau, China.
The Chinese Loess Plateau region has long been suffering from serious soil erosion. Thus, large-scale afforestation has continued during the past decades in order to control soil erosion. Afforestation can dramatically alter nutrient cycles, affect soil-carbon storage, and change hydrology. However, it is unknown how afforestation influences species diversity of the soil seed bank and understory vegetation compared with spontaneous restoration of abandoned land. Forest land with trees planted 30 years ago, abandoned slope land restored spontaneously for 30 years, and the corresponding slopes with remnant natural vegetation were selected as sampling sites. The species richness both in the soil seed bank and vegetation was significantly higher on the afforested slope compared to the spontaneously restored abandoned land. The species similarity between the afforested slope and the remnant slope land was high both in the soil seed bank and standing vegetation compared to the abandoned land. The soil seed bank density varied from 1778 ± 187 to 3896 ± 221 seeds/m², and more than half of it was constituted by annual and biennial species, with no significant difference among sampling habitats. However, the afforested slope had higher seed density of grass and shrub/subshrubs compared to the abandoned slope. The present study indicates that in the study region, characterized by serious soil erosion, afforestation can better facilitate vegetation succession compared to spontaneously restoration of abandoned slope land.


The biological performance of cell-containing phospholipid polymer hydrogels in bulk and microscale form.
The biological performances of a cell-containing phospholipid polymer hydrogel in bulk and miniaturized formats without an additional culture medium support were investigated and compared. The cell-containing hydrogel was formed spontaneously when solutions of commercial polyvinyl alcohol (PVA) and the phospholipid polymer poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-vinylphenylboronic acid (VPBA)] (PMBV) suspended with cells in a cell culture medium are mixed together. Bulk and miniaturized hydrogels, with approximate thicknesses of 3.1 mm and 400 μm, respectively, were prepared in a 96-well microplate and a glass microchip, respectively. In both cases, the hydrogels were homogeneous, and cells were spatially encapsulated. The long-term observation (4 and 8 days) of cell morphology suggested that cells were passively attached to the interface of the hydrogel but were unable to spread and flatten, which inhibited cell growth in both hydrogels. Viability evaluations revealed that cells in both hydrogel formats maintained the same high viability levels after long-term encapsulation. Cytotoxicity assays indicated that the cells in the miniaturized hydrogel maintained a high degree of correlation in cytotoxic sensitivity with the cells in the bulk hydrogel and a routine medium culture. The PMBV/PVA hydrogel not only provides a beneficial cytocompatible microenvironment for long-term cell survival without an additional culture medium support but also creates a static condition for cell sustainment in a microchip similar to that in bulk. The uniform long-term performances of PMBV/PVA hydrogels in bulk and miniaturized formats make them ideal for the development of long-term, flexible, three-dimensional, living cell-based tools for routine cell-based assays and applications on bulk to microscale levels.


Revision Surgery for a Failed Artificial Disc.
This study aimed to present our experience with failures in C-TDR and revision surgery outcomes. We retrospectively examined patients who underwent revision surgery due to the failure of C-TDR between May 2005 to March 2019. Thirteen patients (8 males and 5 females) were included in this study. The mean age was 46.1 years (range: 22-61 years), and the average follow-up period was 19.5 months (range: 12-64 months). The outcome measures of pre- and post-operative neck and arm pain using a visual analogue scale (VAS) and functional impairment were assessed using a modified Japanese Orthopedic Association (JOA) scale and the Neck Disability Index (NDI). The main complaints of patients were posterior neck pain (77%), radiculopathy (62%), and/or myelopathy (62%). The causes of failure of C-TDR were improper indications for the procedure, osteolysis and mobile implant use, inappropriate techniques, and postoperative infection. The most common surgical level was C5-6, followed by C4-5. After revision surgery, the neck and arm pain VAS (preoperative vs. postoperative: 5.46 vs. 1.31; 4.86 vs. 1.08), a modified JOA scale (14.46 vs. 16.69), and the NDI (29.77 vs. 9.31) scores were much improved. C-TDR is good surgical option. However, it is very important to adhere to strict surgical indications and contraindications to avoid failure of C-TDR. The results of reoperations were good regardless of the approach. Therefore, various reoperation options could be considered in patients with failed C-TDR.


Umbilical artery flow velocity waveforms following maternal exercise.
The effect of acute exercise on pregnant women in the last trimester of pregnancy and the changes in fetal umbilical artery flow velocity waveforms were studied. One hundred and forty-three healthy pregnant women underwent an acute exercise test. The exercise was performed on an upright bicycle ergometer for 5 minutes, followed by 30 minutes of recovery. The baseline control values of umbilical artery velocity waveforms were compared with those after exercise. The mean exercise intensity achieved was 67.4% (55-90%) of submaximal exercise. Maternal heart rate rose significantly from a mean of 89 (68-120) beats per minute at rest to a mean of 144 (116-200) beats per minute after exercise (P<0.001). Maternal blood pressure rose significantly after exercise from a mean systolic of 112 (80-150) mmHg to a mean of 147 (120-200) mmHg (P<0.001), and from a mean diastolic of 67 (40-85) mmHg to a mean of 84 (70-100) mm Hg (P<0.001). The mean fetal umbilical pulsatility index (PI) was shown to decrease significantly following exercise at 2 and 4 minutes of recovery. It is concluded that (1) an acute exercise test of this intensity causes a significant decrease in PI index after 2 and 4 minutes of recovery; (2) an exercise intensity between 55 and 75% causes a decrease in the PI after exercise while exercise of 76-90% of submaximal exercise causes a rebound increase after an initial decrease at 2 minutes; (3) it may be possible to use this pattern of exercise as an exercise test to differentiate between low- and high-risk pregnancies. Studies on high-risk pregnancies are being prepared for publication.


Rapid preparative separation of six bioactive compounds from Gentiana crassicaulis Duthie ex Burk. using microwave-assisted extraction coupled with high-speed counter-current chromatography.
A rapid method combining microwave-assisted extraction (MAE) and high-speed counter-current chromatography (HSCCC) was applied for preparative separation of six bioactive compounds including loganic acid (I), isoorientin-4'-O-glucoside (II), 6'-O-β-d-glucopyranosyl gentiopicroside (III), swertiamarin (IV), gentiopicroside (V), sweroside (VI) from traditional Tibetan medicine Gentiana crassicaulis Duthie ex Burk. MAE parameters were predicted by central composite design response surface methodology. That is, 5.0 g dried roots of G. crassicaulis were extracted with 50 mL 57.5% aqueous ethanol under 630 W for 3.39 min. The extract (gentian total glycosides) was separated by HSCCC with n-butanol/ethyl acetate/methanol/1% acetic acid water (7.5:0.5:0.5:3.5, v/v/v/v) using upper phase mobile in tail-to-head elution mode. 16.3, 8.8, 12., 25.1, 40.7, and 21.8 mg of compounds I-VI were obtained with high purities in one run from 500 mg of original sample. The purities and identities of separated components were confirmed using HPLC with photo diode array detection and quadrupole TOF-MS and NMR spectroscopy. The study reveals that response surface methodology is convenient and highly predictive for optimizing extraction process, MAE coupled with HSCCC could be an expeditious method for extraction and separation of phytochemicals from ethnomedicine.


Two genetically distinct lineages of the sika deer, Cervus nippon, in Japanese islands: comparison of mitochondrial D-loop region sequences.
To investigate genetic diversity among populations of the sika deer, Cervus nippon, nucleotide sequences (705-824 bases) of the mitochondrial D-loop regions were determined in animals from 13 localities in the Japanese islands. Phylogenetic trees constructed by the sequences indicated that the Japanese sika deer is separated into two distinct lineages: the northern Japan group (the Hokkaido island and most of the Honshu mainland) and the southern Japan group (a part of the southern Honshu mainland, the Kyushu island, and small islands around the Kyushu island). All sika deer examined in this study shared four to seven units of repetitive sequences (37 to 40 bases each) within the D-loop sequences. The number of tandem repeats was different among the populations, and it was specific to each population. Six or seven repeats occurred in populations of the northern Japan group, while four or five repeats occurred in populations of the southern Japan group. Each repeat unit included several nucleotide substitutions, compared with others, and 26 types were identified from 31 animals. Sequences of the first, second, and third units in arrays were clearly different between the northern and the southern groups. Based on these D-loop data, colonization and separation of the sika deer populations in the Japanese islands were estimated to have occurred less than 0.5 million years before present. Our results provide an invaluable insight into better understanding the evolutionary history, phylogeny, taxonomy, and population genetics of the sika deer.


Strain-specific clearance of seed-dependent tau aggregation by lithium-induced autophagy.
Different conformational strains of tau have been implicated in the clinicopathological heterogeneity of tauopathies. In this study, we hypothesized that distinct strains are degraded in a different manner. Lithium, a drug for bipolar disorder, had previously been reported to reduce aggregation-prone protein content by promoting autophagy. Here, we assessed the effects of lithium on tau aggregates using different tauopathy brain seeds. SH-SY5Y cells were transfected with C-terminal tau fragment Tau-CTF24 (residues 243-441), and Alzheimer's disease (AD), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD) brain seeds were introduced. After 48-h lithium treatment, sarkosyl-insoluble fractions were prepared. Lithium treatment was found to reduce the amount of insoluble tau and p62, and increase LC3-II levels along with the number of autophagic vacuoles in AD-seeded cells. The effects were lower in case of CBD seeds, and comparable between PSP and AD seeds. An inhibitor of myo-inositol monophosphatase (IMPase) also demonstrated similar effects. Overall, the study suggested that aggregated tau protein is degraded by lithium-induced autophagy, influencing IMPase in a strain-specific manner.


Carbon and nitrogen assimilation and partitioning in soybeans exposed to low root temperatures.
Low root temperature effects on vegetative growth of soybean (Harosoy 63 x Rhizobium japonicum USDA 16) were examined in 35 day old plants exposed to temperatures of 15 degrees C (shoots at 25 degrees C) for an 11 day period. Duing this period various aspects of C and N assimilation and partitioning were monitored including shoot night and nodulated root respiration, C and N partitioning to six plant parts, C(2)H(2) reduction, H(2) evolution, leaf area, transpiration, net photosynthesis, and N(2) fixation. The low temperature treatment resulted in a decrease in the net rate of N(2) fixation but nitrogenase relative efficiency increased. In response, the plant retained N in the tissues of the nodulated root and decreased N partitioning to young shoot tissues, thereby inducing the remobilization of N from older leaves, and reducing leaf area development. The leaf area specific rate of net photosynthesis was not affected over the study period; however, shoot and nodulated root respiration declined. Consequently, C accumulated in mature leaves and stems, partly in the form of increased starch reserves. Three possibilities were considered for increasing low temperature tolerance in nodulated soybeans: (a) decrease in temperature optima for nitrogenase, (b) increased development of nodules and N(2) fixation capacity at low temperature, and (c) alterations in the pattern of C and N partitioning in response to low temperature conditions.


[Study on the histochemical staining of boric acid].
The detection of boric acid in the tissue is of significance in investigating its toxicity. Because of this, we have devised a histochemical staining method to detect the presence of boric acid. The outline of this method follows. Frozen 12-14 microns sections, cut by a cryostat, are fixed in anhydrous ethanol and stained for 20 minutes in a protonated curcumin solution. Washing in acetic acid follows, and a red stain results if boric acid is present. This method causes a reaction, in which rosocyanin is formed by the reaction of boric acid and the protonated curcumin, and this principle is now used when an analysis of boric acid is needed. As to procedure, a 1 N concentration of sodium hydroxide is dropped onto a part of the stain to be tested, and the presence of rosocyanin is confirmed if the stain turns blue. Consequently, this staining confirms the presence of boric acid.


Short-term outcomes and major barriers in the management of convulsive status epilepticus in children: a study in Georgia.
Convulsive status epilepticus is the most common childhood neurological emergency in developing countries, where poor healthcare organisation could play a negative role in the management of the condition. Unavailability of second-line injectable anticonvulsants is an additional hindering factor in Georgia. This report reflects the results of the first study aimed at evaluating the epidemiological features of convulsive status epilepticus, as well as identifying obstacles influencing the management of patients with convulsive status epilepticus in Georgia. A prospective, hospital-based study was performed. Paediatric patients with convulsive status epilepticus, admitted to the emergency department of a referral academic hospital from 2007 to 2012, were included in the study. Forty-eight paediatric patients admitted to hospital met the criteria for convulsive status epilepticus. Seizure duration was significantly shorter among the group with adequate and timely pre-hospital intervention. Moreover, patients with appropriate pre-hospital treatment less frequently required mechanical ventilation (p=0.039). Four deaths were detected during the follow-up period, thus the case fatality rate was 8%. Only 31% of patients received treatment with intravenous phenytoin. The study results show that adequate and timely intervention could improve outcome of convulsive status epilepticus and decrease the need for mechanical ventilation. Mortality parameters were comparable to the results from other resource-limited countries. More than one third of patients did not receive appropriate treatment due to unavailability of phenytoin.


Cytokine storm release syndrome and the prospects for immunotherapy with COVID-19, part 4: The role of JAK inhibition.
This review focuses on an alternative strategy utilizing small molecules to inhibit a key signal-transduction pathway, the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway. The JAK-STAT pathway mediates biologic activity for a large number of inflammatory cytokines and mediators.


Does maintenance azithromycin reduce asthma exacerbations? An individual participant data meta-analysis.
Preventing exacerbations is an important goal of asthma treatment. Long-term treatment with azithromycin may help achieve this. Our aim was to conduct a systematic review and individual participant data (IPD) meta-analysis to examine the efficacy of azithromycin in reducing exacerbations in asthma, and in the subphenotypes of noneosinophilic asthma, eosinophilic asthma and severe asthma. We completed a systematic search of Embase, MEDLINE, PubMed, Cochrane Library, ClinicalTrials.gov and reference lists of previous systematic reviews in February 2019. We included parallel-group, double-blind, randomised controlled trials in adults comparing at least 8 weeks of azithromycin treatment with placebo, where the outcome of exacerbations was assessed over at least 6 months. Data were extracted from published sources, Cochrane Risk of Bias Tool was applied and IPD were sought from authors. Reviews were undertaken in duplicate. We conducted an IPD meta-analysis on the primary outcome of exacerbations and a random effects meta-analysis for secondary outcomes. Three studies were identified (n=604). In the IPD meta-analysis, treatment with azithromycin was associated with a reduced rate of exacerbations (oral corticosteroid course due to worsening asthma, antibiotic use for lower respiratory tract infection, hospitalisation and/or emergency department visits) in asthma as well as in the noneosinophilic, eosinophilic and severe asthma subgroups. Examining each exacerbation type separately, patients with eosinophilic asthma reported fewer oral corticosteroid courses, and patients with noneosinophilic and severe asthma reported fewer antibiotic courses. Azithromycin was well tolerated. Maintenance use of azithromycin reduces exacerbations in patients with eosinophilic, noneosinophilic and severe asthma.


Role of Vitamin K in Intestinal Health.
Intestinal diseases, such as inflammatory bowel diseases (IBDs) and colorectal cancer (CRC) generally characterized by clinical symptoms, including malabsorption, intestinal dysfunction, injury, and microbiome imbalance, as well as certain secondary intestinal disease complications, continue to be serious public health problems worldwide. The role of vitamin K (VK) on intestinal health has drawn growing interest in recent years. In addition to its role in blood coagulation and bone health, several investigations continue to explore the role of VK as an emerging novel biological compound with the potential function of improving intestinal health. This study aims to present a thorough review on the bacterial sources, intestinal absorption, uptake of VK, and VK deficiency in patients with intestinal diseases, with emphasis on the effect of VK supplementation on immunity, anti-inflammation, intestinal microbes and its metabolites, antioxidation, and coagulation, and promoting epithelial development. Besides, VK-dependent proteins (VKDPs) are another crucial mechanism for VK to exert a gastroprotection role for their functions of anti-inflammation, immunomodulation, and anti-tumorigenesis. In summary, published studies preliminarily show that VK presents a beneficial effect on intestinal health and may be used as a therapeutic drug to prevent/treat intestinal diseases, but the specific mechanism of VK in intestinal health has yet to be elucidated.


Controlled clinical trial of zolpidem for the treatment of insomnia associated with attention-deficit/ hyperactivity disorder in children 6 to 17 years of age.
The goal was to evaluate the hypnotic efficacy of zolpidem at 0.25 mg/kg per day (maximum of 10 mg/day), compared with placebo, in children 6 through 17 years of age who were experiencing insomnia associated with attention-deficit/hyperactivity disorder. An 8-week, North American, multicenter, double-blind, placebo-controlled, parallel-group study was conducted. Patients underwent stratification according to age (6-11 years [N = 111] or 12-17 years [N = 90]) and were assigned randomly to receive treatment with the study drug or placebo (in a 2:1 ratio). The primary efficacy variable was latency to persistent sleep between weeks 3 and 6. Secondary efficacy variables also were assessed, and behavioral and cognitive components of attention-deficit/hyperactivity disorder were monitored. Safety was assessed on the basis of reports of adverse events, abnormal laboratory data, vital signs, and physical examination findings. The potential for next-day residual effects also was assessed. The baseline-adjusted mean change in latency to persistent sleep at week 4 did not differ significantly between the zolpidem and placebo groups (-20.28 vs -21.27 minutes). However, differences favoring zolpidem were observed for the older age group in Clinical Global Impression scores at weeks 4 and 8. No next-day residual effects of treatment were associated with zolpidem, and no rebound phenomena occurred after treatment discontinuation. Central nervous system and psychiatric disorders were the most-frequent treatment-emergent adverse events (>5%) that were observed more frequently with zolpidem than with placebo; these included dizziness, headache, and hallucinations. Ten (7.4%) patients discontinued zolpidem treatment because of adverse events. Zolpidem at a dose of 0.25 mg/kg per day to a maximum of 10 mg failed to reduce the latency to persistent sleep on polysomnographic recordings after 4 weeks of treatment in children and adolescents 6 through 17 years of age who had attention-deficit/hyperactivity disorder-associated insomnia.


Impact of HIV and AIDS on the eldery: a case study of Chiladzulu district.
HIV may affect the elderly in a number of ways. They may become infected themselves; their children may suffer prolonged illness and incapacity, and need the elderly to look after them; the same children may then die from the illness, leaving the elderly without the support of the next generation; this bereavement also leaves the older people to meet funeral costs and then to take care of orphans left behind. HIV has impacts on old people in ways that are social, economic, psychological and physical in nature. We wanted to understand the socio-economic impacts of HIV/ AIDS on lives of older people following the death of their productive children, and to examine how the elderly struggle to take care of HIV/AIDS orphans. While it is generally appreciated that older people undergo such hardships, little research has been done in Malawi to unravel the magnitude of thes problems. We hope that our findings will increase awareness and lead to more being done to alleviate the problems faced by older people as a result of HIV/AIDS. The results reveal that older people have no alternative but to provide care and support to their ill children, and following the death of their children to become parents again in taking care of the orphaned grandchildren. This is a particularly big challenge in a country like Malawi, dependent on hand-hoeing for subsistence agriculture, where food production requires people who are physically strong. 79% of older people taking care of HIV/AIDS patients or orphans were found to have limited or no information about HIV/AIDS. 31% were dependent on relatives for support, while 69% sustained themselves through piece work, small business or farming. Elderly people need more resources and assistance to enable them to fulfil their vital role in caring for HIV-AIDS orphans. They are also in critical need of more information about HIV-AIDS, as many are sexually active with multiple partners, and preventive information is currently not reaching this important section of the community.


Low vitamin D, but not tobacco use or high BMI, is associated with long-term disability progression in multiple sclerosis.
Low vitamin D levels, tobacco use and high body mass index (BMI) have been linked to adverse disease outcomes in multiple sclerosis (MS), but their influence on long-term disability progression remains unclear. Therefore, we explored whether these modifiable lifestyle factors were associated with 10-year clinical disability progression in patients with MS. In this prospective study, a cohort of 88 patients with relapsing-remitting MS completed a randomized controlled study on ω-3 fatty acids between 2004 and 2008. During 24 months, serum 25-hydroxyvitamin D (25(OH)D), serum cotinine (nicotine metabolite), and BMI were repeatedly measured. In 2017, a follow-up study was conducted among 80 of the participants, including disability assessment by the Expanded Disability Status Scale (EDSS). Linear regression was used to explore associations between the lifestyle factors and the EDSS change over 10 years. Higher seasonally adjusted 25(OH)D levels were associated with lower 10-year EDSS progression (change in EDSS per 1 SD increase in 25(OH)D in a model adjusted for sex, age and baseline EDSS: -0.45 point, 95% CI: -0.75 to -0.16, p=0.003). Further adjustments for potential confounders related to lifestyle and disease status gave similar results. The association was mainly driven by low 25(OH)D levels during spring, as well as seasonally adjusted levels below 80 nmol/L. No clear association was found for BMI and cotinine. Lower 25(OH)D levels, but apparently not tobacco use or higher BMI, were significantly associated with worse long-term disability progression in MS.


The challenges of collecting data on race and ethnicity in a diverse, multiethnic state.
Race and ethnicity are commonly used predictor variables in medical and public health research. Including these variables has helped researchers to describe the etiology of certain disease states. Including race and ethnicity in research has been hypothesis generating in terms of the relationship between genetic and environmental factors in the development of disease. Eliminating health disparities among different racial and ethnic groups has become a national priority. However, incorporating race and ethnicity into health research is complex because these variables are difficult to define and individuals often identify with more than one race or ethnicity. As a "minority-majority", multiethnic, multiracial state, Hawai'i faces unique challenges in incorporating race and ethnicity into research. As the demographics of the United States continue to evolve, many of the challenges faced in Hawai'i will apply to the United States as a whole.


Clearance of Shigella flexneri carriers in a zoologic collection of primates.
In 1990, a high prevalence of Shigella flexneri was documented in the gibbon population at the National Zoological Park. The enzootic strain had been determined to be resistant to most antibiotics and had been responsible for animal deaths. Prevalence of Shigella spp was high in the entire primate population. To address the clinical problem, eradication was attempted by use of enrofloxacin. Facilities disinfection comprised a large part of Shigella clearance efforts. After the clearance protocol was completed gibbons, vs other primate species, were closely monitored for 16 months by subjecting their feces to bacteriologic culturing. The program was successful, with eradication of the problematic strain of S flexneri. However, 10 to 12 months after completion of treatments, S sonnei was isolated from feces of 3 animals living in the same housing facility.


Description of two new species of Rissoella Gray, 1847 (Mollusca, Gastropoda, Heterobranchia) from Venezuela, with a key to the Caribbean species known for the genus.
Two new species of the genus Rissoella Gray, 1847 are described from Venezuela, one from the National Park Morrocoy, Rissoella morrocoyensissp. n. and the other from the Wildlife Refuge Isla de Aves, Rissoella venezolanicolasp. n.Rissoella morrocoyensissp. n. has a deep umbilicus (partly closed), preumbilical cord, black head, hypobranchial gland marked by a pale yellow boomerang-shaped ribbon and it lives on the leaves of the seagrass Thalassia testudinum Banks & König, 1805. Rissoella venezolanicolasp. n. has an angled preumbilical cord which extends to the columella delimiting a trapezoid, a hypobranchial gland marked by a yellow quaver-shaped ribbon and protoconch with fuchsia highlights. It lives on the brown alga Dictyota spp. The records of Rissoella in the Caribbean are revised and illustrations, a comparative table and a key to the Caribbean species known for the genus are provided.


Blind prediction of charged ligand binding affinities in a model binding site.
Predicting absolute protein-ligand binding affinities remains a frontier challenge in ligand discovery and design. This becomes more difficult when ionic interactions are involved because of the large opposing solvation and electrostatic attraction energies. In a blind test, we examined whether alchemical free-energy calculations could predict binding affinities of 14 charged and 5 neutral compounds previously untested as ligands for a cavity binding site in cytochrome c peroxidase. In this simplified site, polar and cationic ligands compete with solvent to interact with a buried aspartate. Predictions were tested by calorimetry, spectroscopy, and crystallography. Of the 15 compounds predicted to bind, 13 were experimentally confirmed, while 4 compounds were false negative predictions. Predictions had a root-mean-square error of 1.95 kcal/mol to the experimental affinities, and predicted poses had an average RMSD of 1.7Å to the crystallographic poses. This test serves as a benchmark for these thermodynamically rigorous calculations at predicting binding affinities for charged compounds and gives insights into the existing sources of error, which are primarily electrostatic interactions inside proteins. Our experiments also provide a useful set of ionic binding affinities in a simplified system for testing new affinity prediction methods.


G-POEM in patients with gastroparesis - gambling for healing or bigger armamentarium ? A case series and review of the literature.
Approximately 30% of all affected patients suffering from gastroparesis do not respond to any available treatment modality. Gastric peroral endoscopic myotomiy (G-POEM, antropyloromyotomy) represents a new principle of therapy. In this single center study, G-POEM showed a high technical success rate with a very low procedural complication rate. However, the clinical response beyond a short-term post-interventional improvement did not succeed in a single patient. The heterogeneity of the clinical picture, which represents a spectrum of different pathophysiological, etiological and clinical characteristics, still requires a therapy tailored to the individual patient. G-POEM should be considered especially in patients with pylorus-dominant gastroparesis.


Tracing cell wall biogenesis in intact cells and plants : selective turnover and alteration of soluble and cell wall polysaccharides in grasses.
Cells of proso millet (Panicum miliaceum L. cv Abarr) in liquid culture and leaves of maize seedlings (Zea mays L. cv LH51 x LH1131) readily incorporated d-[U-(14)C]glucose and l-[U-(14)C]arabinose into soluble and cell wall polymers. Radioactivity from arabinose accumulated selectively in polymers containing arabinose or xylose because a salvage pathway and C-4 epimerase yield both nucleotide-pentoses. On the other hand, radioactivity from glucose was found in all sugars and polymers. Pulse-chase experiments with proso millet cells in liquid culture demonstrated turnover of buffer soluble polymers within minutes and accumulation of radioactive polymers in the cell wall. In leaves of maize seedlings, radioactive polymers accumulated quickly and peaked 30 hours after the pulse then decreased slowly for the remaining time course. During further growth of the seedlings, radioactive polymers became more tenaciously bound in the cell wall. Sugars were constantly recycled from turnover of polysaccharides of the cell wall. Arabinose, hydrolyzed from glucuronoarabinoxylans, and glucose, hydrolyzed from mixed-linkage (1-->3, 1-->4)beta-d-glucans, constituted most of the sugar participating in turnover. Arabinogalactans were a large portion of the buffer soluble (cytoplasmic) polymers of both proso millet cells and maize seedlings, and these polymers also exhibited turnover. Our results indicate that the primary cell wall is not simply a sink for various polysaccharide components, but rather a dynamic compartment exhibiting long-term reorganization by turnover and alteration of specific polymers during development.


Aqueous Two-Phase System Rehydration of Antibody-Polymer Microarrays Enables Convenient Compartmentalized Multiplex Immunoassays.
Multiplex immunoassays are rapidly increasing in popularity due to the offered advantages of increased throughput and decreased sample volume requirements. However, a major weakness inherent to multiplex enzyme-linked immunosorbent assays (ELISA) is generation of false signals through reagent-driven cross-talk. Typically, multiplex platforms necessitate bath application of antibody cocktails, increasing probability of nonspecific antibody binding, especially when multiplexing large numbers of analytes. Aqueous two-phase systems (ATPS) exploiting the phase-separating polymers poly(ethylene) glycol (PEG) and dextran (DEX) have been used to compartmentalize antibodies and prevent cross-talk in multliplex, plate-based ELISA. However, the resulting protocol is tedious and lengthy, and requires too many user steps to be practical for widespread use. Here, we report an improved, user-friendly, cross-talk-free multiplex ELISA method in which dehydrated arrays of colocalized capture and detection antibodies in DEX are prepared on multiwell plates. Addition of a PEG-based sample buffer rehydrates antibody/DEX droplets for analysis. In this report, we demonstrate rehydrated ATPS components for multiplex ELISA retain the ability to compartmentalize antibodies and prevent cross-talk, while analytes in sample buffer partition into rehydrated DEX droplets for analysis. Utility of this method was demonstrated through successful quantitative analysis of five inflammatory cytokines in lipopolysaccharide-stimulated ThP-1 cell culture supernatant.


Measurement of sarcoplasmic reticulum Ca2+-ATPase activity and E-type Mg2+-ATPase activity in rat heart homogenates.
The presence of a high and nonlinear Ca2+-independent (or basal) ATPase activity in rat heart preparations makes difficult the reliable measurement of sarcoplasmic reticulum (SR) Ca2+-ATPase activity by usual methods. A spectrophotometric assay for the accurate determination of SR Ca2+-ATPase activity in unfractionated homogenates from rat heart is described. The procedure is based on that reported by Simonides and van Hardeveld (1990, Anal. Biochem. 191, 321-331) for skeletal muscle homogenates. To avoid overestimation of the Ca2+-ATPase activity of cardiac homogenates that occurs when sequential measurements of total and basal ATPase activities are performed, two parallel and independent assays are required: one with low (micromolar) and other high (millimolar) calcium concentration. Addition of thapsigargin (0.2 microM) blocked totally the activity considered as Ca2+-ATPase activity. Using this method, the rat heart homogenate Ca2+-ATPase activity was 10.5 +/- 2.0 micromol. min-1 x g-1 tissue wet weight (n = 8). Likewise, a spectrophotometric assay for measuring E-type Mg2+-ATPase activity in cardiac total homogenates has been developed, comparing the following characteristics of the enzymatic activity in homogenate and a membrane-enriched fraction: first-order rate constant for ATP-dependent inactivation, Km for ATP, and effects of concanavalin A, Triton X-100, and specific inhibitors.


Is bronchial hyperresponsiveness more frequent in women than in men? A population-based study.
To test the hypothesis that a greater proportion of women than men react to methacholine challenge and investigate the possible reasons for any differences observed, we recruited 495 subjects 20 to 44 yr of age (50.9% male) in Paris and 304 subjects (51.3% male) in Montpellier (France), as part of the European Community Respiratory Health Survey. The proportion of responders (PD20 < or = 4 mg methacholine) was 33.7% in women and 11.9% in men (odds ratio = 3.8; 95% confidence interval = 2.4-6.0) in Paris and 43.2% in women and 29.5% in men (odds ratio = 1.8; 95% confidence interval = 1.1-2.9) in Montpellier. These differences could not be explained by asthma, respiratory symptoms, atopy, or lung function parameters. In stepwise logistic regressions including sex, asthma, and asthma-like symptoms, nasal allergies, atopy, baseline FEV1, FEV1%pred, FVC, and FEV1%FVC, the odds-ratios for the effect of female sex on PD20 < or = 4 mg methacholine were 5.2 (3.0-9.0) in Paris and 2.2 (1.2-3.8) in Montpellier. Reacting to low doses of methacholine (PD20 < or = 0.5 mg) was associated with asthma and atopy in both men and women. In contrast, reacting to doses between 0.5 and 4 mg was associated with asthma and atopy only in men and with heavy tobacco consumption only in women. We conclude that the excess of hyperresponsiveness in women is not due to their having smaller lung size or airway caliber than men and may be related to a greater susceptibility to smoking.


Aurora kinase A is essential for correct chromosome segregation in mouse zygote.
Aurora-A kinase (AURKA), a member of the serine/threonine protein kinase family, is involved in multiple steps of mitotic progression. It regulates centrosome maturation, mitotic spindle formation, and cytokinesis. While studied extensively in somatic cells, little information is known about AURKA in the early cleavage mouse embryo with respect to acentrosomal spindle assembly. In vitro experiments in which AURKA was inactivated with specific inhibitor MLN8237 during the early stages of embryogenesis documented gradual arrest in the cleavage ability of the mouse embryo. In the AURKA-inhibited 1-cell embryos, spindle formation and anaphase onset were delayed and chromosome segregation was defective. AURKA inhibition increased apoptosis during early embryonic development. In conclusion these data suggest that AURKA is essential for the correct chromosome segregation in the first mitosis as a prerequisite for normal later development after first cleavage.


Association between posttraumatic stress disorder and nonfatal drug overdose.
North America is in the midst of a growing drug overdose crisis. While prescription opioid misuse and synthetic opioids such as fentanyl have been implicated in the overdose crisis, less attention has been given to the role that posttraumatic stress disorder (PTSD) may play in this crisis. As such, this study sought to examine the relationship between PTSD and risk of nonfatal overdose among people who use drugs (PWUD). Data were derived from three prospective cohorts of PWUD in Vancouver, Canada. For each participant, PTSD was assessed using the PTSD Checklist for the DSM-5. Multivariate logistic regression analysis was used to estimate the relationship between PTSD and nonfatal overdose, adjusting for potential confounders. Between 2016 and 2018 among 1,059 PWUD, including 363 (34%) nonmale participants, 171 (16%) experienced a nonfatal drug overdose in the past 6 months, and 414 (39%) met criteria for a provisional PTSD diagnosis. In multivariate analysis, PTSD (adjusted odds ratio = 1.98, 95% confidence interval [1.4, 2.79]) remained independently associated with nonfatal overdose after adjustment for a range of confounders. Among participants in these community-recruited cohorts of PWUD, having a provisional PTSD diagnosis nearly doubled the risk of nonfatal overdose. The findings from this study support the need to incorporate a trauma-informed approach within the current overdose prevention framework. Education and training relating to trauma and PTSD should be prioritized for health care professionals who work with and treat PWUD. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Significance of LINC00460 in the progression and prognosis in digestive tract tumors.
The long intergic non-protein coding RNA 460 (LINC00460) is abnormally highly expressed in gastrointestinal tumors and plays an important role in promoting tumor formation and development. LINC00460 is mainly distributed in cytoplasm and has many abnormal gene variants of single nucleotide polymorphism in tumors. LINC00460 can promote the proliferation, metastasis, angiogenesis, radiotherapy and chemotherapy resistance, inhibit the apoptosis of tumor cells, and further promote the malignant progression of tumors via involving in chromatin state maintenance, methylation modification, endogenous competition and transcriptional regulation. It may serve as a valuable tumor marker and therapeutic target.


Immunogenicity of amodiaquine in the rat.
Amodiaquine is an antimalarial drug that has been associated with adverse reactions which may be immune mediated. Specific IgG anti-amodiaquine antibodies were detected after administration of the drug to rats (269 mumols/kg for 4 days), using an enzyme-linked immunosorbent assay employing amodiaquine conjugated to metallothionein as an antigen. A positive immune response was observed regardless of the route of administration, but the magnitude of the response in terms of antibody titre was in the order intraperitoneal administration greater than intramuscular administration greater than oral administration. Hapten inhibition experiments with structurally related drugs defined the specificity of the antibody which appears to recognize a conjugate of amodiaquine quinone imine and cysteine residues present in protein. Amodiaquine was converted to a protein-reactive species by activated human polymorphonuclear leucocytes in vitro, and this may provide a mechanism for immunogen formation in vivo. A humoral immune response was observed with doses of amodiaquine that did not produce either direct hepatotoxicity or leucopenia. Thus an animal model has been developed with which to investigate the toxicological consequences of amodiaquine immunogenicity.


Massively parallel computation of absolute binding free energy with well-equilibrated states.
A force field formulator for organic molecules (FF-FOM) was developed to assign bond, angle, and dihedral parameters to arbitrary organic molecules in a unified manner including proteins and nucleic acids. With the unified force field parametrization we performed massively parallel computations of absolute binding free energies for pharmaceutical target proteins and ligands. Compared with the previous calculation with the ff99 force field in the Amber simulation package (Amber99) and the ligand charges produced by the Austin Model 1 bond charge correction (AM1-BCC), the unified parametrization gave better absolute binding energies for the FK506 binding protein (FKBP) and ligand system. Our method is based on extensive work measurement between thermodynamic states to calculate the free energy difference and it is also the same as the traditional free energy perturbation. There are important requirements for accurate calculations. The first is a well-equilibrated bound structure including the conformational change of the protein induced by the binding of the ligand. The second requirement is the convergence of the work distribution with a sufficient number of trajectories and dense spacing of the coupling constant between the ligand and the rest of the system. Finally, the most important requirement is the force field parametrization.


Hybrid Ablation for Atrial Fibrillation: Safety & Efficacy of Unilateral Epicardial Access.
Hybrid ablation combines thoracoscopic epicardial ablation with percutaneous catheter based endocardial ablation for the treatment of AF. The purpose of this study was to evaluate the safety and efficacy of hybrid ablation surgery for the treatment of atrial fibrillation (AF), and to compare outcomes of unilateral vs bilateral thoracoscopic epicardial ablation. Patients with documented AF who underwent hybrid ablation were followed post-operatively for major events. Major events were classified into 2 categories consisting of (1) safety, comprising all-cause mortality and major morbidities, and (2) efficacy, which included recurrence of atrial arrhythmia, cessation of antiarrhythmic drugs (AAD), and completeness of lesion set. A total of 84 consecutive patients were consented for hybrid ablation. Patients presented with an average AF duration of 85.9 months before hybrid ablation. 80 patients underwent successful thoracoscopic epicardial ablation. At 1-year, 87% (60/69) of patients were free from AF and 73% (50/69) were free from AF and off AAD. 63 patients completed both epicardial and endocardial hybrid ablation with posterior wall isolation achieved in 89% (56/63) of patients. Unilateral epicardial ablation was associated with significantly shorter hospital length of stay compared to bilateral surgical approached (3.9 vs 6.7 days, p = 0.002) with no difference in freedom from AF between groups at 1 year. Hybrid ablation for atrial fibrillation is effective for patients at high risk for recurrence after catheter ablation. The unilateral surgical approach may be associated with shorter hospital stay with no appreciable effect on procedure success rates.