Activation of the nuclear transcription factor kappaB (NFkappaB) and differential gene expression in U87 glioma cells after exposure to the cytoprotector amifostine.
Amifostine has been approved as a therapy to decrease the incidence of moderate-to-severe xerostomia in patients undergoing postoperative radiation treatment for head-and-neck cancer. As a reducing agent capable of participating in intracellular reductive/oxidative processes, it has the potential to affect redox-sensitive transcription factors and gene expression. Amifostine's active free thiol WR-1065 was investigated to determine its effect on nuclear transcription factor kappaB (NFkappaB) activation and subsequent gene expression in U87 glioma cells. The human glioma cell line U87 was grown to confluency and then exposed to WR-1065 at a concentration of 40 microM for times ranging from 30 min to 24 h. Changes in cell cycle were monitored by flow cytometry. The effect of WR-1065 on NFkappaB activation was determined by a gel shift assay. Changes in gene expression as a function of time of exposure to WR-1065 were determined by Northern blot and the Atlas Human cDNA Expression Array (Clontech, Palo Alto, CA). Changes in gene expression using the Atlas Array were verified by reverse transcriptase-polymerase chain reaction (RT-PCR) with gene-specific primers. Exposure of U87 cells to 40 microM WR-1065 resulted in a marked activation of NFkappaB between 30 min and 1 h after treatment. Expression of MnSOD, an NFkappaB-responsive gene, was enhanced by over 2-fold after 16 h of treatment and remained elevated at 24 h. During this period of time, no changes in cell cycle distribution were observed. To assess changes in the expression levels of NFkappaB-responsive genes as a function of WR-1065 exposure, cDNA arrays containing 49 genes identified as having DNA-binding motifs for NFkappaB were used. Only five genes were found to be significantly affected at 1, 4, and/or 16 h of treatment. GST-3 and c-myc were repressed up to 2- and 4-fold, respectively. The expression levels of IL-2Ra, RANTES, and c-myb, in contrast, were enhanced up to 14-, 3-, and 2-fold, respectively. The remaining genes having NFkappaB-responsive elements in their promoter regions were either not expressed (20 genes) or were not affected (24 genes) by exposure to WR-1065. The redox-sensitive transcription factor NFkappaB can be activated in U87 glioma cells by the active thiol form of the cytoprotector amifostine. Activation of NFkappaB by the antioxidant WR-1065 is accompanied by a reduced expression of the oncogene c-myc and an enhanced expression of the antioxidant gene MnSOD, a gene whose expression in tumor cells is relatively low, but when overexpressed has been correlated with a suppression of the malignant phenotype. Activation of NFkappaB by WR-1065, however, results in selective rather than global changes in the expression of genes containing NFkappaB-responsive elements.


Regional distribution of 203PbCl2 in the mouse after intravenous injection.
Lead is a known neurotoxicant, and concentrations of lead in the CNS after acute exposure to low doses have not been studied in detail. In this investigation, the temporal distribution of lead ([203Pb]), injected intravenously at no-carrier-added (NCA) (0.6-0.7 mumol/kg) and at carrier-added (7.0-62.3 mumol/kg) levels was determined in mice. Concentrations of [203Pb] were measured in major organs and in different regions of the brain. Ex-vivo autoradiographic visualization was used to confirm and extend the brain distribution findings. The highest concentrations of NCA [203Pb] were observed initially in the kidneys (52% of the injected dose (ID)/g at 1 hr) and liver (10.5% ID/g at 30 min). Uptake into bone occurred gradually, reaching 25% ID/g at 24 hr. In accord with previous reports, excretion of the tracer was very slow. Approximately 80% of total ID remained in the body after 24 hr and 68% at 48 hr. Interestingly, in the mouse brain, the highest levels of [203Pb] were noted in the area of the hypothalamus. At all times between 30 min and 16 hr postinjection, and at all Pb dose levels injected, the accumulation of [203Pb] in the hypothalamic region exceeded that in all other brain regions examined. Autoradiography performed at the 16 hr time point confirmed the high uptake and strong retention of [203Pb] by the hypothalamus. These studies afford new insight into the distribution of acutely administered lead in the brain, and may have implications for the understanding of some of the neurotoxic effects of lead.


KTI11 and KTI13, Saccharomyces cerevisiae genes controlling sensitivity to G1 arrest induced by Kluyveromyces lactis zymocin.
The Kluyveromyces lactis zymocin and its gamma-toxin subunit inhibit cell cycle progression of Saccharomyces cerevisiae. To identify S. cerevisiae genes conferring zymocin sensitivity, we complemented the unclassified zymocin-resistant kti11 and kti13 mutations using a single-copy yeast library. Thus, we identified yeast open reading frames (ORFs) YBL071w-A and YAL020c/ATS1 as KTI11 and KTI13 respectively. Disruption of KTI11 and KTI13 results in the complex tot phenotype observed for the gamma-toxin target site mutants, tot1-7, and includes zymocin resistance, thermosensitivity, hypersensitivity to drugs and slow growth. Both loci, KTI11 and KTI13, are actively transcribed protein-encoding genes as determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and in vivo HA epitope tagging. Kti11p is highly conserved from yeast to man, and Kti13p/Ats1p is related to yeast Prp20p and mammalian RCC1, components of the Ran-GTP/GDP cycle. Combining disruptions in KTI11 or KTI13 with a deletion in TOT3/ELP3 coding for the RNA polymerase II (RNAPII) Elongator histone acetyltransferase (HAT) yielded synthetic effects on slow growth phenotype expression. This suggests genetic interaction and possibly links KTI11 and KTI13 to Elongator function.


A novel CCM2 variant in a family with non-progressive cognitive complaints and cerebral microbleeds.
Lobar cerebral microbleeds are most often sporadic and associated with Alzheimer's disease. The aim of our study was to identify the underlying genetic defect in a family with cognitive complaints and multiple lobar microbleeds and a positive family history for early onset Alzheimer's disease. We performed exome sequencing followed by Sanger sequencing for validation purposes on genomic DNA of three siblings with cognitive complaints, reduced amyloid-beta-42 in CSF and multiple cerebral lobar microbleeds. We checked for the occurrence of the variant in a cohort of 363 patients with early onset dementia and/or microbleeds. A novel frameshift variant (c.236_237delAC) generating a premature stop codon in the CCM2 gene shared by all three siblings was identified. Pathogenicity of the variant was supported by the presence of cerebral cavernous malformations in two of the siblings and by the absence of the variant exome variant databases. Two siblings were homozygous for APOE-ϵ4; one heterozygous. The cognitive complaints, reduced amyloid-beta-42 in CSF and microbleeds suggest preclinical Alzheimer's disease, but the stability of the cognitive complaints does not. We hypothesize that the phenotype in this family may be due to a combination of the CCM2 variant and the APOE status. © 2016 The Authors. American Journal of Medical Genetics Part B: Neuropsychiatric Genetics Published by Wiley Periodicals, Inc.


Salivary Biomarkers in Periodontitis Patients: A Pilot Study.
Forty subjects were included: 10 periodontally healthy subjects and 30 periodontitis patients. Periodontal examination and saliva sampling were performed in all patients. Levels of salivary cytokines including IL-1β, IL-6, MMP-8, and IL-10 were evaluated by a sandwich ELISA test kit. Data were analyzed by SPSS for Windows. Regarding individual biomarkers, IL-1β, IL-6, and MMP-8 levels were significantly higher in periodontitis patients (p ≤ 0.001, p < 0.05, respectively). The concentration of these proteins in saliva showed a significant association with gingival index and pocket depth measurements and may reflect the clinical status of healthy and diseased periodontium. However, no significant differences were observed for the IL-10 component. IL-1β and IL-6 concentrations were statistically higher in periodontitis patients and may be used as potential tools in periodontitis diagnosis.


Purification and properties of a membrane-bound phospholipase B from baker's yeast (Saccharomyces cerevisiae).
Phospholipase B bound tightly to the membrane fraction of baker's yeast (Saccharomyces cerevisiae) was purified approximately 226-fold by extraction with sodium deoxycholate (DOC), acetone precipitation, ammonium sulfate fractionation, and column chromatographies on Phenyl-Sepharose CL-4B, DEAE-Sepharose CL-6B, and Sepharose 4B. Only one major activity peak with an apparent molecular weight of 330,000 was detected by gel filtration on Sepharose 4B. However, two glycoprotein bands, one major and one minor, were evident on SDS-polyacrylamide gel electrophoresis in the absence of 2-mercaptoethanol. Isoelectric focusing also revealed two activities, pI 3.4 (major) and pI 3.0 (minor). The purified enzyme had phospholipase B activity (in the presence of DOC) and lysophospholipase activity in a ratio of 1:4.8. Both activities had an optimum pH of 3.5-4.0. Phospholipase B activity was appreciably stimulated only by DOC among bile acids, but lysophospholipase activity was markedly inhibited by them. Both activities were not stimulated by Ca2+ and inhibited by SDS, Triton X-100, Fe3+ and Al3+. The purified enzyme hydrolyzed the acyl ester bonds of phosphatidylcholine sequentially, first the 2-acyl and then the 1-acyl groups. It preferentially hydrolyzed 1-acyl-sn-glycero-3-phosphocholine and, to a lesser degree, 2-acyl-sn-glycero-3-phosphocholine.


Evaluation Of Anti-inflammatory Mediated Anti-hemorrhoidal Activity of Lawsonia inermis on Croton Oil Induced Hemorrhoidal Rats.
Hemorrhoids are a common recto-anal disorder commonly known as piles or tissue clumps in the rectum. In normal individuals, they were known as anal cushions. In the anus, they are composed of rectal blood vessels, muscles, and elastic fibres. When bulged, it can cause bleeding, constipation, itching, severe pain, and bleeding in the anus. Inflammation of the anal cushion remains major pathogenesis for the development of hemorrhoids. Inflammatory mediators like neutrophils, TNF-α, and IL-6 seem to play a major role in the development of disease. This study aims to carry out the ethanolic leaf extract of Lawsonia inermis (L. inermis) and evaluate its anti-hemorrhoidal activity both in vitro and in vivo. Furthermore, Molecular Docking was performed on the crystal structure of COX-2 with the selective compound 23d-(R) (PDB ID: 3NTG) protein. The current study is to estimate an anti-inflammatory mediated anti-hemorrhoidal activity of ethanolic leaf extract of L. inermis at different doses of 200 mg/kg/ir and 400 mg/kg/ir in croton oil-induced hemorrhoidal rats. Pilex ointment is taken as a reference standard in the present study. Evan's blue extravasation technique were applied in the study to quantify the proinflammatory protein. From the study results, a dose-dependent effect was found for ethanolic leaf extract of Lawsonia inermis at 200 mg/kg and 400 mg/kg causing a significant reduction of serum Proinflammatory mediators TNF-α, IL-6, and plasma neutrophils in croton oil-induced hemorrhoidal rats. Studies represented that the plant extract can significantly inhibit protein denaturation of egg albumin in in vitro and found to reduce croton oil induced inflammatory mediators in hemorhhoidal rats.


NANOG: a promising target for digestive malignant tumors.
NANOG has been extensively researched since its discovery by Chambers et al. NANOG is a homeodomain transcription factor and an essential regulator of embryonic stem cell (ESC) self-renewal, which inhibits differentiation. Cancer stem cells (CSCs) are a small subset of cells that are thought to drive uncontrolled tumor growth; CSCs retain the tumor capabilities of self-renewal and propagation. The existence of CSCs was recently shown by direct experimental evidence. NANOG is expressed in CSCs and ESCs, although it remains unclear whether ESCs and CSCs share similar mechanisms in the regulation of physical and biological processes. Several studies suggest that the expression level of NANOG is high in cancer tissues and low or absent in normal tissues. High levels of NANOG expression are associated with advanced stages of cancer and a poor prognosis, indicating that it plays a vital role in tumor transformation, tumorigenesis, and tumor metastasis. NANOG is part of a complex regulatory network that controls cell fate determination, proliferation, and apoptosis. NANOG cooperates with other regulators, such as microflora, transcription factors, and kinases, in cancer cells. NANOG might have a promising future in anti-cancer and other therapeutic treatments, which could improve human health.


Root-Lesion Nematodes Affecting Dryland Cereals in the Semiarid Pacific Northwest U.S.A.
Root-lesion nematodes (Pratylenchus spp.) are parasites that invade and deteriorate roots, thereby reducing the efficiency of water and nutrient uptake. Pratylenchus neglectus and P. thornei are the two species that are most prevalent and cause reduced yields of rainfed wheat and barley in semiarid regions of the Pacific Northwest. They are particularly damaging where wheat and barley are produced without irrigation in areas receiving less than 450 mm (18 in.) of precipitation annually. This review is focused on the biology and management of P. neglectus and P. thornei in semiarid rainfed agriculture. Characteristics of climates, soils, and crop production systems are described as a preface to constraints placed upon management options. Discussions include the economic importance, host ranges, and protocols for sampling and species identification. Discussion of disease management options include crop rotation, genetic resistance and tolerance, planting date, trap and biofumigant crops, crop nutrition, chemical and biological nematicides, and tillage. Predictions for rainfed agriculture in a period of changing climate are presented, as are suggestions for important areas of research including crop genetics, nematode testing, and communication of results, Pratylenchus biology, mechanisms of resistance, the phytobiome, and closing the "yield gap" between actual and attainable yields.


Clinical Characteristics and Risk of Relapse for Patients with Stage I-II Diffuse Large B-cell Lymphoma Treated in First Line with Immunochemotherapy.
Diffuse large b-cell lymphoma (DLBCL) is an aggressive and potentially curable lymphoma that presents itself as stage I-II in 30% of all cases. It is known that in these localized stages, 15-20% of patients treated without rituximab eventually relapse, but less data exist regarding rituximab era. We have analyzed clinico-pathological features and risk of relapse in 98 patients with I-II stage DLBCL in complete response (CR) or unconfirmed CR (CRu) after first-line treatment consisting of immunochemotherapy. Twelve patients (12.2%) eventually relapsed. Late relapse, more than two years after diagnosis, occurred in three patients, and early relapse, less than two years after diagnosis, was documented in nine patients. Median time from diagnosis to relapse was 0.61 years for patients with early relapse and 3.66 years for patients with late relapse. The second CR rate obtained was similar in the late and in early relapsing patients, being 33% versus 44% (p = 0.072), respectively. Three-year overall survival (OS) was 22% for early relapsing patients and 33% for late relapsing patients (p = 0.65). In conclusion, patients who are diagnosed with stage I-II DLBCL and achieve a CR/CRu with first line immunochemotherapy have a good prognosis. However, a proportion of patients relapse, and this is less frequent in patients treated with first line with immunochemotherapy. These patients have a poor prognosis.


[FACTORS PREDICTING OVERALL AND DISEASE-FREE SURVIVAL AFTER SURGERY FOR INVASIVE BLADDER CANCER].
To evaluate the prognosis of patients who underwent surgery for invasive bladder cancer and to search for prognostic factors. The files of all the patients who underwent radical or partial cystectomy for invasive bladder cancer between 1992 and 2014 were reviewed. The effect of various prognostic factors was evaluated by uni- and multivariate analyses. A total of 160 patients were included in the study and were followed for a median period of 25.5 months after surgery. The overall 2 years and 5 years survival rates were 70% and 61.2% respectively. The disease-free 2 years and 5 years survival rates were 64.4% and 61.9% respectively. The overall 2 years and 5 years survival rates of patients with disease limited to the bladder (≥T2N0) were 88.2% and 82.4% and of patients with disease extending beyond the bladder (≤T3N0) 56.5% and 45.7% respectively. Factors that were found to be significantly associated with overall survival were: TNM stage, co-morbidity (Charlson 6-11) and the tumor's diameter. No association was found between: disease presentation, smoking habits, positive cytology, the tumor being primary or secondary, variant histology, the presence of endophytic growth pattern, the presence of CIS, hydronephrosis, positive lymph nodes on pre-operative imaging, surgery type (radical or partial cystectomy) and adjuvant chemotherapy. The survival rates of the locally treated patients match the reported rates in the literature. Tumors' T stage were found to be the strongest prognostic factor. Tumors' diameter was found to be an independent prognostic factor. This is reported here for the first time in the literature.


In vivo significance of T cells in the development of Coxsackievirus B3 myocarditis in mice. Immature but antigen-specific T cells aggravate cardiac injury.
Acute myocardial damage similar to that seen in human myocarditis occurs in BALB/c mice after infection with coxsackievirus B3 (CB3) or encephalomyocarditis virus (EMC). To investigate the role of antigen-specific T cells in the pathogenesis of this disorder, we compared CB3 disease expression in T cell-deficient, athymic nude (nu/nu) mice, in heterozygote (nu/+) mice with normal T cell function, and in nu/nu mice reconstituted with spleen cells from CB3- or EMC-infected nu/+ mice. Acute myocarditis occurred in both nu/nu and nu/+ mice. Severe myocarditis, however, developed only in nu/+ and nu/nu mice reconstituted with CB3-sensitized T cells, but not in those reconstituted with EMC-sensitized T cells. Myocardial virus titer and serum anti-CB3 antibody production were similar in nu/+ and nu/nu groups. Additionally, the presence of Thy 1.2 (pan T), Ly 1 (precursor of other T cell subsets), and Ly 2 (suppressor/cytotoxic T) positive cells was demonstrated in the myocardium in nu/+ and nu/nu mice reconstituted with CB3-sensitized T cells, but not with T cells sensitized by another virus, EMC. These results indicate that immature but antigen-specific T cells play a role in the pathogenesis of ongoing myocarditis.


The effects of yellow fever immunization (17DD) inadvertently used in early pregnancy during a mass campaign in Brazil.
This study describes the consequences of the inadvertent immunization of pregnant women during a mass vaccination campaign in the Campinas region of the state of São Paulo, Brazil, in February and March 2000. The study was carried out by the Women's Comprehensive Healthcare Center (CAISM), at the State University of Campinas (UNICAMP). The objective of the study was to evaluate the possible effects of the vaccine on pregnancy and conceptus, and to assess congenital infection resulting from immunization. Pregnant women who received the YF vaccine were identified at primary health clinics and referred to the study site, a public reference, high-risk clinic, serving 42 towns in a region with a population of 3,000,000. A 12-month serological follow-up for newborns (PRNT), and an examination to detect congenital abnormalities was offered to pregnant women, who signed a consent form. In a sub-sample of women who were delivered at the study site, additional exams were proposed: neonatal fontanel ultrasound, funduscopy, audiometry, neuro-pediatric follow-up until 12 months of age, and IgM detection at birth. Fifteen blood samples from placentas and umbilical cords were tested for PCR. A total of 480 pregnant, immunized women were identified, who had received the vaccine at a mean of 5.7 weeks (95% CI 5.2-6.2) of gestation. The great majority of women were unaware of their pregnancy at the time they were vaccinated, and only 46.7% were counseled to avoid immunization if pregnant. After a minimum 6-week interval, 98.2% pregnant women were IgG positive. A total of 19.6% of women reported mild adverse events (headache, fever or myalgia). No IgM antibodies were detected at birth and no placental or umbilical cord blood was positive according to PCR. The frequency of malformations (2.3% or 7/304 babies), miscarriages (2.5% or 11/441 pregnancies), stillbirths (0.7%) and premature delivery (7.8%) was similar to that found in the general population. At 12 months follow-up, 7% of samples were reactive to PRNT. However, after 12 months, only one child was seropositive. Contrary to a previous study, maternal seroconversion was very high when immunization was carried out in early pregnancy. Vaccine applied during the first trimester does not appear to cause malformations, complications to the central nervous system, nor adverse perinatal results as represented by premature deliveries or perinatal deaths. The 12-month serological follow-up is inconclusive and should be extended to 24 months. Evaluation of the risk of miscarriage was hindered by late presentation at the study clinic.


Limb reconstruction by free-tissue transfer combined with the Ilizarov method.
In four complex cases of extremity reconstruction, we have been able to overcome the problems of combined bone and soft-tissue loss and length discrepancy by a combination of free-tissue transfer and the Ilizarov method of slow distraction. It is our observation that gradual distraction of a free tissue is a safe and viable procedure; the free tissue tolerates the pins of the circular external fixator well, and there is an equal degree of distraction and regeneration of the transferred free tissue and the native recipient tissue without evidence of wound dehiscence. Corticotomy through free tissue and in close proximity to vascularized bone is safe, with the subsequent bone regeneration not unlike that of normal bone. Manipulation by slow distraction does not appear to compromise the vasculature of the recipient bed for later microsurgical procedures or endanger the axial flow pattern of the transferred free tissue.


Oral pamidronate in the prevention and treatment of skeletal metastases in patients with breast cancer.
Bone metastases are common in patients with breast cancer and cause considerable morbidity and deterioration of the quality of life. The main pathogenetic mechanism is stimulation of osteoclastic bone resorption by factors produced by the cancer cells. Pamidronate given intravenously suppresses bone resorption and is an effective treatment of malignancy-associated hypercalcemia. Recent data indicate that it can also reduce skeletal morbidity in normocalcemic patients with breast cancer and osteolytic metastases. In a series of studies we examined the long-term efficacy of oral pamidronate in the prevention and treatment of skeletal metastases in patients with breast cancer. In patients with bone metastases oral pamidronate given for a median period of about 20 months reduced significantly skeletal morbidity and had a favourable effect on selective aspects of the quality of life of the patients. Treatment did not alter the radiological course of the disease or the overall survival of the patients. In contrast, oral pamidronate given to patients with advanced breast cancer but no demonstrable bone metastases did not prevent or delay the appearance of the first clinical or radiological manifestation of bone metastases. This treatment is therefore very effective in patients with established metastatic bone disease. More studies are needed to define the place of pamidronate (and of other bisphosphonates) in the prevention of bone metastases in patients at risk.


Number 2 Feibi Recipe Inhibits H2O2-Mediated Oxidative Stress Damage of Alveolar Epithelial Cells by Regulating the Balance of Mitophagy/Apoptosis.
Reactive oxygen species (ROS)-mediated alveolar epithelial cell (AEC) injury and apoptosis are considered to be the initiating link of idiopathic pulmonary fibrosis (IPF), and protecting AECs can alleviate IPF. This study aimed to explore the protective effect of number 2 Feibi recipe (FBR-2) medicated serum on H2O2-mediated oxidative stress injury in AECs and further explore its mechanism. We found that FBR-2 can regulate downstream antioxidant enzymes expression by activating nuclear factor erythroid 2-related factor 2 (Nrf2), reducing the level of intracellular ROS, protecting mitochondrial function and improving cell survival. FBR-2 can also activate mitophagy through the PINK1/Parkin pathway. Moreover, FBR-2 can inhibit apoptosis by blocking the mitochondrial apoptosis mechanism. In summary, these data indicate that FBR-2 medicated serum can inhibit H2O2-mediated oxidative stress damage in AECs by regulating the balance of mitophagy/apoptosis. This study provides new evidence for the antifibrotic effect of FBR-2 and provides new drug candidates for the clinical treatment of IPF.


Cardiovascular disease in COPD: mechanisms.
It is now well established that cardiovascular disease contributes significantly to both morbidity and mortality in COPD. Shared risk factors for cardiovascular disease and COPD, such as smoking, low socioeconomic class, and a sedentary lifestyle contribute to the natural history of each of these conditions. However, it is now apparent that alternative, novel mechanisms are involved in the pathogenesis of cardiovascular disease, and these may play an important role in driving the increased cardiovascular risk associated with COPD. In this article, we discuss the potential mechanisms that link COPD to an increased risk of cardiovascular disease.


Temperature-induced phase transitions in the rock-salt type SiC: a first-principles study.
The phase transitions in the rock-salt type SiC (B1-SiC) under decompression are studied in the framework of first-principles molecular dynamics simulations up to room temperature. The transformation pathways were determined based on an analysis of the symmetry and phonon spectra of high-symmetry transient structures identified in the simulations. The plausible pathways of the transformation of B1-SiC into the 3C-, 2H-, 4H-, 12R-SiC polytypes were suggested. The transformation paths were found to depend on both the availability of soft phonon modes in an unreconstructed phase and the initial conditions of the simulation. It is shown that an increase in cell volume at decompression leads to the condensation of a certain phonon mode. As a result, an intermediate state forms due to the atomic displacements and to subsequent strains related to this mode. All the decompressed structures were compressed back under pressure of 120-250 GPa depending on the type of the decompressed phase and simulation temperature that was in the range of 300-1200 K. The suggested scheme of structural identification can be used to determine the transition paths for the structural transformations of other similar structures under pressure.


Characterization of globin domains: heme binding to the central exon product.
We have prepared and isolated the peptide fragments coded for by the three exons of the human beta-globin gene, using the arginine-specific protease clostripain (EC 126.96.36.199). The region encoded by the central exon (amino acid residues 31-104) contains an arginine at position 40. This site was less susceptible to cleavage than the two sites that correspond to the exon-intron boundaries, and the isolated central fragment was an approximately equimolar mixture of the entire central fragment, beta(o) (31-104), and the somewhat smaller fragment contained within it, beta(o) (41-104). This central fragment mixture bound heme stoichiometrically and tightly at micromolar concentrations, generating a strong Soret absorption band as well as a characteristic absorption band in the visible spectrum. The Soret band occurred at the same wavelength and had the same shape as in hemoglobin, exhibiting an intensity greater than (2/3) that achieved when native intact beta globin is reconstituted with heme. Nearly the full intensity was regained when an equivalent of heme was added to the unfractionated digest, suggesting that the noncovalently associated side fragments add precision to the fit of the heme pocket. Three controls were used in establishing the specificity of heme binding to the central fragment mixture. Similar, but preliminary, experiments have also been undertaken with alpha globin. A clostripain digest containing alpha(o) (1-31) and alpha(o) (32-141) bound heme, yielding a Soret band identical to that observed in alpha subunits reconstituted from the native globin chains and heme. Measurements of circular dichroism spectra as indices of secondary structure suggested a role for the side exon products in the acquisition of the native three-dimensional structure of hemoglobin. These experiments confirm a prediction of W. Gilbert that the product of the central exon of the globin gene is a complete functional domain that binds heme tightly and specifically.


Effect of insulin on caveolin-enriched membrane domains in rat liver.
Compartmentalization of signaling molecules may explain, at least in part, how insulin or growth factors achieve specificity. Caveolae/rafts are specialized lipid compartments that have been implicated in insulin signaling. In the present study, we investigated the role of caveolin-enriched membrane domains (CMD) in mediating insulin signaling in rat liver. We report the existence of at least two different populations of CMD in rat liver plasma membranes (PM). One population is soluble in Triton X-100 and seems to be constitutively associated with cytoskeletal elements. The other population of CMD is located in a membrane compartment insoluble in Triton X-100 with light buoyant density and is hence designated CMD/rafts. We found evidence of rapid actin reorganization in rat liver PM in response to insulin, along with the association of CMD/rafts and insulin signaling molecules with a cell fraction enriched in cytoskeletal elements. The presence of CMD in liver parenchyma cells was confirmed by the presence of caveolin-1 in primary rat hepatocyte cultures. Cholesterol depletion, effected by incubating hepatocytes with 2 mm methyl-beta-cyclodextrin, did not permeabilize the cells or interfere with clathrin-dependent internalization. However, at this concentration, methyl-beta-cyclodextrin perturbed CMD of hepatocyte PM and inhibited insulin-induced Akt activation and glycogen synthesis but did not affect insulin-induced insulin receptor kinase tyrosine phosphorylation. These events, together with the presence of a functional insulin receptor in CMD of rat liver PM, suggest that insulin signaling is influenced by the interaction of caveolae with cytoskeletal elements in liver.


Comparison of absorption after inhalation and instillation of uranium octoxide.
Values for the absorption parameters were compared after inhalation or intratracheal instillation of 1.5 microm mass median aerodynamic diameter (MMAD) 233U3O8 particles into the lungs of HMT strain rats. The two sets of parameter values were similar, as were the calculated dose coefficients and predicted biokinetics for workers. Hence the inhalation and instillation techniques can probably both be used to generate values of the absorption parameters for U3O8.


Identification of novel Coxiella burnetii genotypes from Ethiopian ticks.
Coxiella burnetii, the etiologic agent of Q fever, is a highly infectious zoonotic bacterium. Genetic information about the strains of this worldwide distributed agent circulating on the African continent is limited. The aim of the present study was the genetic characterization of C. burnetii DNA samples detected in ticks collected from Ethiopian cattle and their comparison with other genotypes found previously in other parts of the world. A total of 296 tick samples were screened by real-time PCR targeting the IS1111 region of C. burnetii genome and from the 32 positive samples, 8 cases with sufficient C. burnetii DNA load (Amblyomma cohaerens, n = 6; A. variegatum, n = 2) were characterized by multispacer sequence typing (MST) and multiple-locus variable-number tandem repeat analysis (MLVA). One novel sequence type (ST), the proposed ST52, was identified by MST. The MLVA-6 discriminated the proposed ST52 into two newly identified MLVA genotypes: type 24 or AH was detected in both Amblyomma species while type 26 or AI was found only in A. cohaerens. Both the MST and MLVA genotypes of the present work are closely related to previously described genotypes found primarily in cattle samples from different parts of the globe. This finding is congruent with the source hosts of the analyzed Ethiopian ticks, as these were also collected from cattle. The present study provides genotype information of C. burnetii from this seldom studied East-African region as well as further evidence for the presumed host-specific adaptation of this agent.


Flt-3 ligand increases microchimerism but can prevent the therapeutic effect of donor bone marrow in transiently immunosuppressed cardiac allograft recipients.
C3H (H2k) mice received 50 x 10(6) B10 (H2b) bone marrow (BM) cells either alone or with flt-3 ligand (FL) (10 microg/day), tacrolimus (2 mg/kg/day), or both agents for 7 days. Donor MHC class II+ (IAb+) cells were quantitated in spleens by immunohistochemical analysis, and donor class II DNA detected in BM by PCR. Donor cells were rare in the BM alone and BM + FL groups, whereas there was a substantial increase in chimerism in the BM + tacrolimus group. Addition of FL to BM + tacrolimus led to a further eightfold increase in donor cells and enhanced donor DNA compared with the BM + tacrolimus group. This increase in donor cells was almost 500-fold compared with BM alone. C3H recipients of B10 heart allografts given perioperative B10 BM and tacrolimus (days 0-13) exhibited a markedly extended median graft survival time (MST, 42 days) compared with those given tacrolimus alone (MST, 22 days). Addition of FL (10 microg/day; 7 days) to BM + tacrolimus prevented the beneficial effect of donor BM (MST, 18 days). BM alone or BM + FL resulted in uniform early heart graft failure (MST < 8 days). Functional studies revealed maximal antidonor MLR and CTL activities in the BM- and BM + FL-treated groups, with minimal activity in the tacrolimus-treated groups. Thus, dramatic growth factor-induced increases in chimerism achieved under cover of immunosuppression may result in augmented antidonor T cell reactivity and reduced graft survival after immunosuppressive drug withdrawal. With FL, this may reflect striking augmentation of immunostimulatory dendritic cells.


Deletion variants of RABGAP1L, 10q21.3, and C4 are associated with the risk of systemic lupus erythematosus in Korean women.
Several copy number variations (CNVs) have been found to be associated with systemic lupus erythematosus (SLE) through the target gene approach. However, genome-wide features of CNVs and their role in the risk of SLE remain unknown. The aim of this study was to identify SLE-associated CNVs in Korean women. Genome-wide assessments of CNVs were performed in 382 SLE patients and 191 control subjects, using an Illumina HumanHap610 BeadChip genotyping platform. SLE-associated CNV regions that were identified by genome-wide association study (GWAS) were replicated in quantitative polymerase chain reaction (PCR) and deletion-typing PCR analyses in an independent sample set comprising 564 SLE patients and 511 control subjects. Of 144 common CNV regions, 3 deletion-type CNV regions in 1q25.1, 8q23.3, and 10q21.3 were found to be significantly associated with SLE by GWAS analysis. In the independent replication, the CNV regions in 1q25.1 (RABGAP1L) and 10q21.3 were successfully replicated (odds ratio [OR] 1.30, P=0.038 and OR 1.90, P=3.6×10(-5), respectively), and the associations were confirmed again by deletion-typing PCR. The CNV region in the C4 gene, which showed a potential association in the discovery stage, was included in the replication analysis and was found to be significantly associated with the risk of SLE (OR 1.88, P=0.01). Through deletion-typing PCR, the exact sizes and breakpoint sequences of the deletions were defined. Individuals with the deletions in all 3 loci (RABGAP1L, 10q21.3, and C4) had a much higher risk of SLE than did those without any deletions in the 3 loci (OR 5.52, P=3.9×10(-4)). These CNV regions can be useful to identify the pathogenic mechanisms of SLE, and might be used to more accurately predict the risk of SLE by taking into consideration their synergistic effects on disease susceptibility.


Experimental closed system surgical procedures and intraocular pressure fluctuation.
To determine the behavior of intraocular pressure and its relationship with infusion pressure in vitrectomy, phacofragmentation and phacoemulsification surgeries in rabbits. Intraocular pressure fluctuation was measured in 24 eyes of 12 rabbits submitted to vitrectomy, phacofragmentation and phacoemulsification procedures (eight eyes per group). The procedures were divided according to the force of aspiration of the instrument. Intraocular pressure was monitored with a computerized polygraph by means of a cannula introduced into the vitreous chamber. Intraocular pressure showed a mean variation from 33 mmHg (maximum) to 6 mmHg (minimum). Vitrectomy showed the greatest difference between weak and strong aspiration. The greatest fluctuations occurred during procedures with strong aspiration, with phacoemulsification showing the widest variation, with maximum peaks almost reaching 50 mmHg. The infusion pressure varied less than the intraocular pressure, especially at the lowest pressures. The fluctuation of intraocular pressure during ocular surgeries was not great. The higher the aspiration pressure, the higher the intraocular pressure. The infusion pressure did not show a good correlation with intraocular pressure.


Alcohol Content in the 'Hyper-Reality' MTV Show 'Geordie Shore'.
To quantify the occurrence of alcohol content, including alcohol branding, in the popular primetime television UK Reality TV show 'Geordie Shore' Series 11. A 1-min interval coding content analysis of alcohol content in the entire DVD Series 11 of 'Geordie Shore' (10 episodes). Occurrence of alcohol use, implied use, other alcohol reference/paraphernalia or branding was recorded. All categories of alcohol were present in all episodes. 'Any alcohol' content occurred in 78%, 'actual alcohol use' in 30%, 'inferred alcohol use' in 72%, and all 'other' alcohol references occurred in 59% of all coding intervals (ACIs), respectively. Brand appearances occurred in 23% of ACIs. The most frequently observed alcohol brand was Smirnoff which appeared in 43% of all brand appearances. Episodes categorized as suitable for viewing by adolescents below the legal drinking age of 18 years comprised of 61% of all brand appearances. Alcohol content, including branding, is highly prevalent in the UK Reality TV show 'Geordie Shore' Series 11. Two-thirds of all alcohol branding occurred in episodes age-rated by the British Board of Film Classification (BBFC) as suitable for viewers aged 15 years. The organizations OfCom, Advertising Standards Authority (ASA) and the Portman Group should implement more effective policies to reduce adolescent exposure to on-screen drinking. The drinks industry should consider demanding the withdrawal of their brands from the show. Alcohol content, including branding, is highly prevalent in the MTV reality TV show 'Geordie Shore' Series 11. Current alcohol regulation is failing to protect young viewers from exposure to such content.


Investigations into economic returns resulting from foliar fungicides and application timing on management of tar spot in Indiana hybrid corn.
Tar spot, caused by Phyllachora maydis, is the most significant yield-limiting disease of corn (Zea mays L.) in Indiana. Currently, fungicides are an effective management tool for this disease and partial returns from their use under different disease severity conditions has not previously been studied. Between 2019 and 2021, two separate field experiments were conducted in each year in Indiana to assess the efficacy of nine foliar fungicide products and nine fungicide application timings based on corn growth stages on tar spot symptoms and stromata, canopy greenness, yield, and influence on partial returns. All fungicides evaluated significantly suppressed tar spot development in the canopy and increased canopy greenness over the nontreated. Additionally, applications of mefentrifluconazole + pyraclostrobin, metconazole + pyraclostrobin, cyproconazole + picoxystrobin at tassel, and propiconazole + benzovindiflupyr + azoxystrobin between the tassel and dough growth stages were the most effective at significantly reducing disease severity, increasing canopy greenness, protecting yield, and offered the greatest partial return. Fungicide products varied in their ability to protect yield under low and high disease severity conditions relative to the nontreated control. Consistently, positive yield increases were observed when disease severity was high which translated to greater profitability relative to low severity conditions. On average, the yield increases across foliar fungicide products and timed application treatments were 544.6 kg/ha and 1,020.7 kg/ha greater and partial returns using a grain value of $0.17/kg were $92.6/ha and $173.5/ha greater, respectively when high severity conditions occurred. This research demonstrates that foliar fungicides and appropriately timed fungicide applications can profitably be used to manage tar spot in Indiana under high disease severity conditions.


New drug delivery strategies targeting the GnRH receptor in breast and other cancers.
Cancer is the uncontrolled division of abnormal cells in a specific organ. Globally, about one in six deaths is due to cancer. Despite the plethora of research being undertaken worldwide to find a cure for cancer, it remains a significant challenge. Cancer targeting via agents designed to interfere with some specifically or highly expressed molecules in cancer cells has been a shift in the treatment of various forms of cancers. The development of drug delivery systems, specifically to cancer cells, is a common approach that succeeded in increasing the efficacy and reducing the side effects of different anticancer agents. Gonadotropin-releasing hormone (GnRH) is a naturally occurring hormone with receptors overexpressed in many types of cancers related or unrelated to the reproductive system. Several drug delivery systems were developed using GnRH derivatives as targeting agents. In this review, we first discuss the role of GnRH and its receptors in cancer. Then, we provide a detailed insight into different delivery systems developed using GnRH derivatives as targeting agents in various types of GnRH receptor overexpressing cancers. Some promising findings from these studies indicate that GnRH receptor targeting is a potential strategy to efficiently guide anticancer therapeutics, diagnostic agents, and nucleic acids directly to cancer cells. Lastly, some limitations of the current research and suggestions for more successful outcomes in clinical trials of these delivery systems are highlighted.


Prescribing patterns of target-specific oral anticoagulants: an academic hospital perspective.
Target-specific oral anticoagulants have been rapidly adopted into clinical practice for stroke prophylaxis and venous thromboembolism treatment, raising concerns about off-label prescribing practices. We conducted a retrospective review of consecutive patients prescribed dabigatran, rivaroxaban or apixaban prior to inpatient hospitalization over an 18-month period to examine the off-label prescribing frequency, contraindications and related complications. Chart review included baseline demographics, hospital admitting service, outpatient prescribing service, renal function, therapeutic indication, echocardiographic findings, contraindications, major bleeding events and vital status. We identified 160 patients who received a target-specific oral anticoagulant prior to hospitalization. Over half (53.1%) of the patients received rivaroxaban, 43.7% received dabigatran and 3.1% received apixaban. Atrial fibrillation (68.1%) and venous thromboembolism treatment (25.6%) were the most common indications. Ninety percent of patients had a U.S. Foods and Drugs Administration (FDA)-approved indication for therapy. Major bleeding events occurred in 4.4% of patients. Cardiology was the most common prescribing and admitting service (43.8 and 31.3%), and more frequently adhered to FDA-approved indications (97 vs. 84%, P = 0.01). There were no significant differences between prescribing services regarding major contraindications (P = 0.14) and major bleeding events (P = 0.77). Off-label prescription rates for target-specific oral anticoagulants were infrequent and not associated with increased adverse events.


Structure-based discovery of clinically approved drugs as Zika virus NS2B-NS3 protease inhibitors that potently inhibit Zika virus infection in vitro and in vivo.
Zika virus (ZIKV) infection may be associated with severe complications in fetuses and adults, but treatment options are limited. We performed an in silico structure-based screening of a large chemical library to identify potential ZIKV NS2B-NS3 protease inhibitors. Clinically approved drugs belonging to different drug classes were selected among the 100 primary hit compounds with the highest predicted binding affinities to ZIKV NS2B-NS3-protease for validation studies. ZIKV NS2B-NS3 protease inhibitory activity was validated in most of the selected drugs and in vitro anti-ZIKV activity was identified in two of them (novobiocin and lopinavir-ritonavir). Molecular docking and molecular dynamics simulations predicted that novobiocin bound to ZIKV NS2B-NS3-protease with high stability. Dexamethasone-immunosuppressed mice with disseminated ZIKV infection and novobiocin treatment had significantly (P < 0.05) higher survival rate (100% vs 0%), lower mean blood and tissue viral loads, and less severe histopathological changes than untreated controls. This structure-based drug discovery platform should facilitate the identification of additional enzyme inhibitors of ZIKV.


[Changes in cell-mediated immune reactions in Crohn's disease].
In a prospective study the effect on cell-mediated immunity (Mérieux-Multitest), nutritional state and parameters of inflammation was compared in conservatively (n = 21) and surgically (n = 20) treated patients with Crohn's disease. Before therapy both groups showed a significantly lower immune response compared with healthy controls. The therapeutical effect was similar in both groups concerning normalization of parameters of inflammation, CDAI and state of nutrition. In contrast, normalization of immune response in skin test was only observed after bowel resection, while conservatively treated patients remained in hyperergy. In conclusion, the impairment of the cell mediated immunity in Crohn's disease must be secondary. Excluding patients with steroid therapy there was no correlation between preoperative annergy or hyperergy and postoperative complications.


Serological differentiation of avian infectious bronchitis field isolates using an enzyme immunoassay: presence of Dutch strains in West Germany.
Six field virus isolates were identified as the etiologic agent of avian infectious bronchitis (IB) in West Germany. A serological comparison was carried out using the M-41 strain from the Massachusetts serotype and the Dutch variant strains D-207, D-1466, D-3128, and D-3896 in a cross indirect immunoperoxidase test. Three isolates demonstrated similarity to the M-41 strain; the remainder showed a closer antigenic relationship to the Dutch variant strains. These results correspond to differences observed in a hemagglutination-inhibition test. The enzyme immunoassay is proposed as another possible method for differentiating IB virus strains.


Perfusion cardiovascular magnetic resonance: Comparison of an advanced, high-resolution and a standard sequence.
Technical advances in perfusion cardiovascular magnetic resonance (CMR), particularly accelerated data acquisition methods, allow myocardial perfusion imaging with unprecedented spatial resolution. However, it is not clear how implementation of these recent advances affects perfusion image quality, signal and contrast to noise ratios (SNR & CNR) and the occurrence of important artefacts in routine clinical imaging. The objective of this study was therefore to compare a standard and an advanced, high-resolution perfusion sequence. A standard ultrafast gradient echo perfusion sequence (st-GrE) was compared with an advanced kt-accelerated steady state free precession sequence (ktBLAST-SSFP) at 1.5 T in healthy volunteers (n = 16) and in patients (n = 32) with known or suspected coronary artery disease. Volunteers were imaged with both sequences at rest and patients underwent stress and rest imaging with either st-GrE or ktBLAST-SSFP prior to X-ray coronary angiography.A blinded expert scored image quality and respiratory artefact severity and also classified patients for the presence of CAD. The extent, transmurality and duration of dark rim artefacts (DRA) as well as signal to noise (SNR) and contrast to noise (CNR) were quantified. In normal hearts ktBLAST-SSFP imaging resulted in significantly improved image quality (p = 0.003), SNR (21.0 ± 6.7 vs. 18.8 ± 6.6; p = 0.009), CNR (15.4 ± 6.1 vs. 14.0 ± 6.0; p = 0.034) and a reduced extent (p = <0.0001) and transmurality (p = 0.0001) of DRA. In patients ktBLAST-SSFP imaging resulted in significantly improved image quality (p = 0.012), and a reduced extent (p = <0.0001), duration (p = 0.004) and transmurality (p = <0.0001) of DRA. Sensitivity and specificity for the detection of CAD against X-ray angiography was comparable with both sequences. There was a non-significant trend towards increased respiratory artefacts with ktBLAST-SSFP in both patients and volunteers. Advanced high resolution perfusion CMR using a k-t-accelerated SSFP technique results in significantly improved image quality, SNR and CNR and a reduction in the extent and transmurality of DRA compared to a standard sequence. These findings support the use of advanced perfusion sequences for clinical perfusion imaging however further studies exploring whether this results in improved diagnostic accuracy are required.


Advances in pediatric MR imaging.
This article describes the considerable technical achievements that have been made in MR imaging in the evaluation of pediatric patients. The latest techniques in improving signal intensity, resolution, and speed are discussed. The multitude of new options for pediatric MR imaging are illustrated, including higher field strength imaging, multi-channel coil technology coupled with parallel imaging, and new pulse sequence designs. Several future directions in the field of pediatric body and musculoskeletal imaging also are highlighted.


Indicators to evaluate quality of care in head and neck cancer in Spain.
This study aimed to develop a set of criteria and indicators to evaluate the quality of care of patients with head and neck cancer (HNC). A systematic literature review was conducted to identify valuable criteria/indicators for the assessment of the quality of care in HNC. With the aid of a technical group, a scientific committee of oncologists specialised in HNC used selected criteria to propose indicators that were evaluated with a two-round Delphi method. Indicators on which consensus was achieved were then prioritised by the scientific committee to develop a final set of indicators. We proposed a list of 50 indicators used in the literature or developed by us to be evaluated with a Delphi method. There was consensus on the appropriateness of 47 indicators in the first round; the remaining 3 achieved consensus in the second round. The 50 indicators were scored to prioritise them, leading to a final selection of 29 indicators related to structure (3), process (22), or outcome (4) and covering diagnosis, treatment, follow-up, and health outcomes in patients with HNC. Easy-to-use index cards were developed for each indicator, with their criterion, definition, formula for use in real-world clinical practice, rationale, and acceptable level of attainment. We have developed a set of 29 evidence-based and expert-supported indicators for evaluating the quality of care in HNC, covering diagnosis, treatment, follow-up, and health outcomes.


[Effect of cyclophosphamide on hematopoietic stem cells in mice with iron overload].
To explore the effect of cyclophosphamide on hematopoietic stem cells (HSCs) in mice with iron overload. Mouse models of iron overload were established in 30 male C57BL/6 mice by intraperitoneal injections of iron dextran at low (0.25 g/kg), moderate (0.5 g/kg), and high (1 g/kg) doses (n=10), with another 10 PBS-treated mice as the control group. The changes in body weight, liver, spleen and bone marrow of the mice were recorded, and serum level of ferritin was detected. The mice receiving a moderate dose of iron dextran were further divided into 8 groups for observation at different time points (D1, D2, D3, D4, D5, D6, D7, and D14 groups) and were given intraperitoneal injection of 50 mg/kg cyclophosphamide (Cy) for 2 consecutive days. Peripheral blood cells, bone marrow mononuclear cells (BMMNCs), and the frequencies of different HSCs (HPCs, HSCs, LT-HSCs) in the BMMNCs were monitored. The cell cycle distribution in the HSCs, level of reactive oxygen species and the microenvironment of the HSCs were analyzed using flow cytometry. Compared with the control mice, the mice with iron overload showed obvious weight loss with significantly increased serum ferritin level, enlargement of the liver and spleen, and iron deposition in the organs (P < 0.05). No significant changes were noted in the peripheral blood of the mice with iron overload. The cyclophosphamide-treated mice exhibited significantly decreased number of WBCs and lymphocyte ratio at days 1 to 4 (P < 0.05). The numbers of BMMNCs and HPCs in mice with iron overload did not show significant changes as compared with those in the control mice, but the numbers of HSCs and LTHSCs decreased significantly in the mice with iron overload (P < 0.05). In cyclophosphamide-treated mice, the number of HSCs increased since day 1 and reached the peak level on day 3 (P < 0.05). Compared with those in the control group, the HSCs did not exhibit significant changes in cell cycle distribution in mice with iron overload, but the proportion of G0/G1 cells decreased significantly in cyclophosphamide group since day 1 and reached the lowest level on day 3 (P < 0.05). Iron deposition in the bone marrow causes long- term damages of the HSCs in the bone marrow but does not induce obvious changes in the peripheral blood. In mice with iron overload, intraperitoneal injection of 50 mg/kg cyclophosphamide for two days promotes cell cycle changes of the resting HSCs to mobilize the HSCs, and this effect is the most obvious on day 4.


Effects of locoweed toxin on rats.
Rats fed pellets containing locoweed developed typical microscopic lesions of locoweed poisoning. Rats fed a water extract of locoweed or urine from sheep given locoweed also developed microscopic lesions. This indicated that the locoweed toxin is water soluble and is excreted in the urine.


The genetics of gout: towards personalised medicine?
Over the last decade, there have been major advances in the understanding of the genetic basis of hyperuricaemia and gout as well as of the pharmacogenetics of urate-lowering therapy. Key findings include the reporting of 28 urate-associated loci, the discovery that ABCG2 plays a central role on extra-renal uric acid excretion, the identification of genes associated with development of gout in the context of hyperuricaemia, recognition that ABCG2 variants influence allopurinol response, and the impact of HLA-B*5801 testing in reducing the prevalence of allopurinol hypersensitivity in high-risk populations. These advances, together with the reducing cost of whole genome sequencing, mean that integrated personalised medicine approaches may soon be possible in clinical practice. Genetic data may inform assessment of disease prognosis in individuals with hyperuricaemia or established gout, personalised lifestyle advice, selection and dosing of urate-lowering therapy, and prevention of serious medication adverse effects. In this article, we summarise the discoveries from genome-wide association studies and discuss the potential for translation of these findings into clinical practice.


Effects of the herbicide isoproturon on metallothioneins, growth, and antioxidative defenses in the aquatic worm Tubifex tubifex (Oligochaeta, Tubificidae).
Metallothioneins (MTs) are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MT contents is considered to be a specific biomarker of metal exposure. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and antioxidative defenses. Therefore, the induction of MTs as biomarkers of exposure to the pesticide isoproturon has been investigated in the aquatic worms Tubifex tubifex. MT levels in exposed worms increased significantly (p < 0.05) after 2, 4, and 7 days of exposure to different concentrations of isoproturon (maximum increase compared to unexposed controls: +148.56% for 10 mg l(-1) after 4 days of exposure). In response to isoproturon, the activity of glutathione-S-transferase (max. +52%), glutathione-reductase (max. +100%), and catalase (max. +117%) increased, demonstrating the occurrence of an oxidative stress response to the herbicide. Thus, the increase in MT contents caused by isoproturon was interpreted as a defense response towards increased oxidative stress generated by the herbicide. Residues of isoproturon and its metabolites, 1-(4-isopropylphenyl)-3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin were detected in the worm growth medium. Half-life of the herbicide was shorter at a low (0.1 mg l(-1)) initial concentration. The herbicide accumulated in T. tubifex but no metabolite could be detected.


Diabetes mellitus promotes susceptibility to periodontitis-novel insight into the molecular mechanisms.
Diabetes mellitus is a main risk factor for periodontitis, but until now, the underlying molecular mechanisms remain unclear. Diabetes can increase the pathogenicity of the periodontal microbiota and the inflammatory/host immune response of the periodontium. Hyperglycemia induces reactive oxygen species (ROS) production and enhances oxidative stress (OS), exacerbating periodontal tissue destruction. Furthermore, the alveolar bone resorption damage and the epigenetic changes in periodontal tissue induced by diabetes may also contribute to periodontitis. We will review the latest clinical data on the evidence of diabetes promoting the susceptibility of periodontitis from epidemiological, molecular mechanistic, and potential therapeutic targets and discuss the possible molecular mechanistic targets, focusing in particular on novel data on inflammatory/host immune response and OS. Understanding the intertwined pathogenesis of diabetes mellitus and periodontitis can explain the cross-interference between endocrine metabolic and inflammatory diseases better, provide a theoretical basis for new systemic holistic treatment, and promote interprofessional collaboration between endocrine physicians and dentists.


Assessing the clinical significance of single items relative to summated scores.
How many items are needed to measure an individual's quality of life (QOL)? This article describes the strengths and weaknesses of single items and summated scores (from multiple items) as QOL measures. We also address the use of single global measures vs multiple subindices as measures of QOL. The primary themes that recur throughout this article are the relationships between well-defined research objectives, the research setting, and the choice single item vs summated scores to measure QOL. The conceptual framework of the study, the conceptual fit with the measure, and the purpose of the assessment should all be considered when choosing a measure of QOL. No "gold standard" QOL measure can be recommended because no "one size fits all." Single items have the advantage of simplicity at the cost of detail. Multiple-item indices have the advantage of providing a complete profile of QOL component constructs at the cost of increased burden and of asking potentially irrelevant questions. The 2 types of indices are not mutually exclusive and can be used together in a single research study or in the clinical setting.


Confocal imaging of mouse mandibular condyle cartilage.
Mice are commonly used to study the temporomandibular joint (TMJ) and to model human TMJ disease. However, evaluating TMJ pathology in mice using standard histologic methods is time consuming, labor intensive, and dependent upon investigators' expertise at consistently orienting and sectioning across tiny specimens. We describe a method that uses confocal microscopy to rapidly and reliably assess indicators of mandibular condyle cartilage pathology in mice. We demonstrate the utility of this method for detecting abnormalities in chondrocyte distribution in mice lacking lubricin (Prg4), the major boundary lubricant of articular cartilage. We further show that the method can provide information about recombination sites and efficiency in mandibular cartilage for Cre-driver strains. Because specimen preparation and data acquisition with confocal microscopy are simple and fast, the method can serve as a primary screening tool for TMJ pathology, before proceeding to complicated, time consuming, secondary analyses.


Selective noradrenaline reuptake inhibitors for schizophrenia.
Schizophrenia is frequently a chronic and disabling illness with a heterogeneous range of symptoms. The positive symptoms usually respond to antipsychotics but the cognitive and negative symptoms of schizophrenia are difficult to treat with conventional antipsychotics and significantly impact on quality of life and social outcomes. Selective noradrenaline reuptake inhibitors (NRIs) increase prefrontal dopamine and noradrenaline levels without significantly affecting subcortical dopamine levels, making them an attractive candidate for treating cognitive and negative symptoms. To investigate the effects of selective noradrenaline reuptake inhibitors (NRIs), compared with a placebo or control treatment, for people with schizophrenia. We searched the Cochrane Schizophrenia Group's Trials Register (up to 7 February 2017) which is based on regular searches of MEDLINE, Embase, CINAHL, BIOSIS, AMED, PubMed, PsycINFO, and registries of clinical trials. There are no language, date, document type, or publication status limitation for inclusion of records into the register. We inspected references of all included studies for further relevant studies. We included randomised controlled trials (RCTs) comparing NRIs with either a control treatment or placebo for people with schizophrenia or related disorders (such as schizoaffective disorder) by any means of diagnosis. We included trials that met our selection criteria and provided useable information. We independently inspected all citations from searches, identified relevant abstracts, and independently extracted data from all included studies. For binary data we calculated risk ratio (RR), for continuous data we calculated mean difference (MD), and for cognitive outcomes we derived standardised mean difference (SMD) effect sizes, all with 95% confidence intervals (CI) and using a random-effects model. We assessed risk of bias for the included studies and used the GRADE approach to produce a 'Summary of findings' table which included our prespecified main outcomes of interest. Searching identified 113 records. We obtained the full text of 48 of these records for closer inspection. Sixteen trials, randomising a total of 919 participants are included. The majority of trials included adults with schizophrenia or similar illness who were inpatients, and while they were poorly characterised, most appeared to include patients with a chronic presentation. The intervention NRI in nine of the 16 trials was reboxetine, with atomoxetine and viloxazine used in the remaining trials. 14 trials compared NRIs with placebo. Only two trials provided data to compare NRIs against an active control and both compared reboxetine to citalopram but at 4 weeks and 24 weeks respectively so they could not be combined in a meta-analysis.One trial was described as 'open' and we considered it to be at high risk of bias for randomisation and blinding, three trials were at high risk of bias for attrition, six for reporting, and two for other sources of bias. Our main outcomes of interest were significant response or improvement in positive/negative mental state, global state and cognitive functioning, average cognitive functioning scores, significant response or improvement in quality of life and incidence of nausea. All data for main outcomes were short term.NRIs versus placeboMental state results showed significantly greater rates of improvement in negative symptoms scores (1 RCT, n = 50; RR 3.17, 95% CI 1.52 to 6.58; very low quality evidence) with NRIs on the PANSS negative. No data were reported for significant response or improvement in positive symptoms, but average endpoint PANSS positive scores were available and showed no difference between NRIs and placebo (5 RCTs, n = 294; MD -0.16, 95% CI -0.96 to 0.63; low-quality evidence). Improvement in clinical global status was similar between groups (1 RCT, n = 28; RR 0.99, 95% CI 0.45 to 2.20; very low quality evidence). Significant response or improvement in cognitive functioning data were not reported. Average composite cognitive scores showed no difference between NRIs and placebo (4 RCTs, n = 180; SMD 0.04, 95% CI -0.28 to 0.36; low-quality evidence). Significant response or improvement in quality of life data were not reported, however average endpoint scores from the GQOLI-74 were reported. Those receiving NRIs had better quality of life scores compared to placebo (1 RCT, n = 114; MD 9.36, 95% CI 7.89 to 10.83; very low quality evidence). All-cause withdrawals did not differ between the treatment groups (8 RCTs, n = 401, RR 0.94 95% CI 0.63 to 1.39; moderate-quality evidence). Rates of nausea were not greater with NRIs (3 RCTs, n = 176; RR 0.49, 95% CI 0.10 to 2.41; low-quality evidence). Our results provide tentative very low quality evidence that compared to placebo, NRIs (specifically reboxetine) may have a benefit on the negative symptoms of schizophrenia. Limited evidence also suggests that NRIs have no effect on the positive symptoms of schizophrenia or cognitive functioning. NRIs appear generally well tolerated with no real differences in adverse effects such as nausea noted between NRIs and placebo. However, these results are based on short-term follow-up and are poor quality - there is need for more good-quality evidence. A large RCT of reboxetine over a longer period of time, focusing specifically on negative and cognitive symptoms as well as more detailed and comprehensive reporting of outcomes, including adverse events, is required.


Workup for Perinatal Stroke Does Not Predict Recurrence.
Perinatal stroke, including neonatal and presumed perinatal presentation, represents the age in childhood in which stroke occurs most frequently. The roles of thrombophilia, arteriopathy, and cardiac anomalies in perinatal ischemic stroke are currently unclear. We took a uniform approach to perinatal ischemic stroke evaluation to study these risk factors and their association with recurrent stroke. We reviewed records of perinatal stroke patients evaluated from August 2008 to February 2016 at a single referral center. Demographics, echocardiography, arterial imaging, and thrombophilia testing were collected. Statistical analysis was performed using Fisher exact test. Across 215 cases, the median follow-up was 3.17 years (1.49, 6.46). Females comprised 42.8% of cases. Age of presentation was neonatal (110, 51.2%) or presumed perinatal (105, 48.8%). The median age at diagnosis was 2.9 days (interquartile range, 2.0-9.9) for neonatal stroke and 12.9 months (interquartile range, 8.7-32.8) for presumed perinatal stroke. Strokes were classified as arterial (149, 69.3%), venous (60, 27.9%), both (4, 1.9%), or uncertain (2, 0.9%) by consensus imaging review. Of the 215 cases, there were 6 (2.8%) recurrent ischemic cerebrovascular events. Abnormal thrombophilia testing was not associated with recurrent stroke, except for a single patient with combined antithrombin deficiency and protein C deficiency. After excluding venous events, 155 patients were evaluated for arteriopathy and cardioembolic risk factors; neither was associated with recurrent stroke. Positive family history of thrombosis was not predictive of abnormal thrombophilia testing. Thrombophilia, arteriopathy, or cardioembolic risk factors were not predictive of recurrent events after perinatal stroke. Thrombophilia evaluation in perinatal stroke should only rarely be considered.


Using measured octanol-air partition coefficients to explain environmental partitioning of organochlorine pesticides.
Octanol-air partition coefficients (Koa) were measured directly for 19 organochlorine (OC) pesticides over the temperature range of 5 to 35 degrees C. Values of log Koa at 25 degrees C ranged over three orders of magnitude, from 7.4 for hexachlorobenzene to 10.1 for 1,1-dichloro-2,2-bis(p-chlorophenyl) ethane. Measured values were compared to values calculated as KowRT/H (where R is the ideal gas constant [8.314 J mol(-1) K(-1)], T is absolute temperature, and H is Henry's law constant) were, in general, larger. Discrepancies of up to three orders of magnitude were observed, highlighting the need for direct measurements of Koa. Plots of Koa versus inverse absolute temperature exhibited a log-linear correlation. Enthalpies of phase transition between octanol and air (deltaHoa) were determined from the temperature slopes and were in the range of 56 to 105 kJ mol(-1) K(-1). Activity coefficients in octanol (gamma(o)) were determined from Koa and reported supercooled liquid vapor pressures (pL(o)), and these were in the range of 0.3 to 12, indicating near-ideal solution behavior. Differences in Koa values for structural isomers of hexachlorocyclohexane were also explored. A Koa-based model was described for predicting the partitioning of OC pesticides to aerosols and used to calculate particulate fractions at 25 and -10 degrees C. The model also agreed well with experimental results for several OC pesticides that were equilibrated with urban aerosols in the laboratory. A log-log regression of the particle-gas partition coefficient versus Koa had a slope near unity, indicating that octanol is a good surrogate for the aerosol organic matter.


Association between loneliness, self-esteem and outcome of life satisfaction in Norwegian adolescents aged 15-21.
Adolescence is an important developmental stage for understanding the role of perceived loneliness and self-esteem on life satisfaction. This study investigated the association between loneliness, self-esteem and the outcome of life satisfaction, as well as potential interaction effects in association with life satisfaction, in a sample of Norwegian adolescents. The study was based on a cross-sectional sample of 1816 adolescents aged 15-21 years. Data were collected in September 2016. The participants reported scores on the five-item Satisfaction with Life Scale, the 10-item Rosenberg Self-Esteem Scale and one item assessing loneliness. Control variables included sex, age, perceived family economy, parents' education, place of birth and perceived bullying. The data were analysed with descriptive and multiple linear regression analysis. A significant negative and moderately strong association was found between loneliness and life satisfaction, where the association was stronger for girls than for boys. Self-esteem showed a significant positive and strong association with life satisfaction; however, no significant interaction effect was found. The findings show the significant role of both loneliness and self-esteem in association with adolescents' perception of life satisfaction. The findings support promoting self-esteem, belongingness and social integration in all daily life contexts for adolescents to support their life satisfaction.


Effect of Anterior Tibial Closing Wedge Osteotomy on Coronal Tibial Alignment in Relation to Preoperative Medial Proximal Tibial Angle and Wedge Height.
The posterior tibial slope has been identified as an anatomic risk factor for anterior cruciate ligament insufficiency and reruptures after anterior cruciate ligament reconstruction. Anterior tibial closing wedge osteotomy for correction of sagittal plane deformities has the potential to cause an unintended change in coronal plane alignment. To evaluate the effects of anterior tibial closing wedge osteotomies for correction of posterior tibial slope on coronal plane alignment using an infratuberosity surgical approach and to identify predictive factors for a change in medial proximal tibial angle (MPTA). Case series; Level of evidence, 4. This study reports on retrospectively obtained data from radiographic measurements of 38 anterior tibial closing wedge osteotomies. All patients underwent revision anterior cruciate ligament reconstruction and had undergone ≥1 previous anterior cruciate ligament reconstruction. In all patients, an infratuberosity approach was used with angular stable plate fixation. Pre- and postoperative radiographs were examined retrospectively to detect changes in the sagittal and coronal plane alignment (posterior tibial slope and MPTA). A multivariate regression analysis was used to identify predictors for a change in MPTA. The study group consisted of 14 women and 24 men whose mean ± SD age at the index procedure was 31.6 ± 8.7 years (range, 17-51 years). Posterior tibial slope decreased significantly (by 7.2° ± 2.3°; P < .001) from 14.6° ± 2.0° preoperatively to 7.4° ± 2.1° postoperatively. MPTA decreased significantly by 1.3° ± 1.5° (P = .005) from pre- to postoperative measurement. Mean wedge height was 9.3 ± 1.1 mm. A lower preoperative MPTA (coefficient = 0.32; P = .017; 95% CI, 0.06-0.59) and larger wedge height (coefficient = 0.48; P = .029; 95% CI, 0.05-0.9) were significant predictive factors for a decrease in MPTA. Anterior tibial closing wedge osteotomy for posterior tibial slope reduction resulted in a slight but significant decrease of the MPTA in the coronal plane. These changes were dependent on the preoperative MPTA and the wedge height.


[Evaluation of Mascia Brunelli rapid antigen test in the diagnosis of group A streptococcal pharyngitis].
Pharyngitis in most cases is due to viral microorganisms however drug therapy without the detection of etiological agent leads to unnecessary use of antibiotics. On the other hand, when the etiologic agent is group A beta-hemolytic streptococci (GAS) it is important to identify the etiologic agent rapidly which will guide the treatment with appropriate antibiotics. The use of highly sensitive rapid tests will contribute significantly to early diagnosis and appropriate therapy. The aim of this study is to evaluate the efficacy of Mascia Brunelli rapid antigen test for the detection of GAS in throat swab samples. A total of 833 throat swab samples submitted to our laboratory with pre-diagnosis of pharyngitis were assessed between June 2016 and August 2016. The samples were simultaneously cultured and tested by rapid Mascia Brunelli Strep-A Card (Mascia Brunelli S.p.a, Italy). For identification, bacitracin sensitivity, PYR test and latex agglutination test in addition to Bruker MALDI-TOF MS (Daltonics, Germany) system were used. The density of GAS growth in the culture was noted. The samples that were false negative with Mascia Brunelli test were re-tested with QuickVue + Strep A Test (Quidel Corporation, San Diego, USA) rapid antigen test. A total of 833 patients, 376 (45.2%) female and 457 (54.8%) male were included in the study. The age range was between 0-94 years with a mean value of 7.86 ± 6.72. 125 (15%) and 94 (11.28%) of the samples were positive with culture and rapid antigen test, respectively. Mascia Brunelli antigen test gave negative results for 31 culture positive samples. Of these 31 samples, 28 were found positive by QuickVue + Strep A antigen test. As a result, the sensitivity of the test was found to be independent of the inoculum effect. The culture positivity rate in patients between 5-15 years was 18.4%. The sensitivity, specificity, positive predictive value, negative predictive value and the accuracy of Mascia Brunelli antigen test, with respect to culture, were 75.2%, 100%, 100%, 95.81% and 96.28%, respectively. In conclusion, the selection of rapid antigen tests with high sensitivity in the diagnosis of GAS pharyngitis will contribute to the prevention of resistance development by appropriate use of antibiotics as well as early diagnosis and appropriate treatment. However, confirmation of negative rapid antigen test results by culture is very important in terms of false diagnosis and prevention of incomplete treatment.


Evolution and differentiation of MSHR gene in different species.
Coat color offers some prospects for evolutionary studies due to its large amount of presumably adaptive coat color variation and conserved genetic mechanisms of generating different coat colors in different species. Melanocyte-stimulating hormone receptor (MSHR) gene is responsible for intraspecific and interspecific color variation in mammals and birds. A total number of 206 MSHR gene sequences belonging to 84 species, 58 genera, and 20 families were analyzed to investigate its evolution and differentiation in different species. Most of the species have 954 bp and stop codon TGA. Species in Callithrix and Callimico have a stop codon mutation from TGA to TGG and elongate 81 bp with TAG as stop codon. Species in Phasianidae, Fringillidae, and Lemuridae also use TAG as stop codon. The Sus scrofa had an insertion of AACCAGACC encoding Asn-Gln-Thr from 85 to 93 bp. In Bovidae, a brown strain of cow with 966 bp due to the 12-bp duplication of GGCATTGCCCGG from 670 to 681 bp encoding for Gly-Ile-Ala-Arg was found. Teiidae has the smallest number of total mutations (6), silent mutations (3), nonsynonymous mutations (3), average number of nucleotide differences (1.519), synonymous nucleotide diversity (pi(s) = 0.0030), and nonsynonymous nucleotide diversity (pi(a) = 0.0029), and Hominidae, Lemuridae, Canidae, and Teiidae have higher ratio of pi(a)/pi(s) (0.537-0.973). The reconstructed phylogenetic tree of MSHR gene of families is basically consistent with the taxonomy of National Center for Biotechnology Information.


Oxidative stress in COPD.
Oxidative stress is now recognized as a major predisposing factor in the pathogenesis of COPD. Existing therapies for COPD are ineffective at halting disease progression, with bronchodilators being the mainstay of pharmacotherapy, providing symptomatic relief only. It is, therefore, important for a better understanding of the underlying mechanisms by which oxidative stress drives disease pathogenesis to develop novel and more effective therapies. Antioxidant capacity in COPD is substantially reduced as a result of cigarette smoking and exacerbations, with oxidative stress persisting long after the cessation of cigarette smoking or exacerbation, due to the continued production of reactive oxygen species from endogenous sources. We discuss (1) how oxidative stress arises in the lung, (2) how it is neutralized, (3) what genetic factors may predispose to the development of COPD, and (4) how this impacts inflammation and autoimmunity in the development of emphysema and small airways disease. Finally, various strategies have been considered to neutralize the increased oxidative burden present in COPD. This review highlights why current antioxidant strategies have so far failed and what promising alternatives are on the horizon. Moreover, a number of studies have shown that there is no single "magic bullet" to combat oxidative stress, but instead a combination therapy, targeting oxidative stress in the various subcellular compartments, may prove to be more effective in COPD.


Domiciliation of Triatoma pseudomaculata (Corrêa e Espínola 1964) in the Jequitinhonha Valley, State of Minas Gerais.
Impact of the vector control program was evaluated eight years after implantation of epidemiological surveillance for Chagas disease in Berilo, a municipality in the Jequitinhonha Valley of the Brazilian State of Minas Gerais. In all 5,242 domiciliary units (96% of the total) were inspected and 10 found to be infested by the triatomine bug Triatoma pseudomaculata. Triatomines were found associated with bats inside one house and in the peridomiciles of the other nine. None of the 111 Triatoma pseudomaculata captured was infected with Trypanosoma cruzi. Noireau et al traps were installed in (n=8) and around (n=100) the infested house but no Trypanpsoma cruzi-infected triatomines were found. None bat, opossums (Didelphis albiventris) and rat captured in the peridomicile were infected with Trypanosoma cruzi although 24% of the inhabitants of the house infested by Triatoma pseudomaculata were seropositive for the parasite, based on ELISA, IHA and IIF.


Concussion in facial trauma patients: a retrospective analysis of 100 patients from a UK major trauma centre.
Concussion is a common and potentially debilitating condition. Research has shown that one-third of patients admitted with facial trauma have concurrent concussion. This study aimed to investigate the burden and management of concussion in patients presenting with acute facial trauma, and to identify potential risk factors within this population. A retrospective observational study was conducted at a UK major trauma centre between 1 January 2019 and 1 February2020. One hundred randomly selected patients who attended the acute clinic responsible for managing facial trauma were identified. No parametric data were included. The Mann-Whitney test was used to detect differences for continuous data, the X2 test for categorical data. Clinical significance was defined as p < 0.05. Forty of 100 patients (40%) had evidence of concussion, of which only 4/40 (10%) had evidence that head injury advice had been given. There was no statistically significant difference between the non-concussed and concussed groups for age (p = 0.145), gender (p = 0.921), mechanism of injury (p = 0.158), or location of facial injury (p = 0.451). Clinical features of concussion were found in 40% of patients suffering from facial injury. Despite this, we found that head injury advice was rarely given. In addition, we identified no risk factors for concussion within this population, highlighting the need to screen all patients who present with facial injury. To improve the identification and management of concussion in these patients, future work should focus on the development of simple screening tools for use in clinic, and the signposting of patients to existing written and online concussion resources.


Effectiveness of individualised intervention on older residents with constipation in nursing home: a randomised controlled trial.
To develop and examine the effectiveness of individualised intervention to reduce constipation among older adults in nursing homes. In long-term care facilities, approximately 60-80% of the residents have symptoms of constipation. Constipation may lead to haemorrhoids, faecal impaction, ulcers, intestinal bleeding and can also lead to a decrease in quality of life. Although a high prevalence of constipation in older adults can be seen, there is a lack of empirical evidence for delivering interventions based on individual risk factors of constipation. Many factors cause constipation but the risk factors are different for each individual. A prospective, randomised control trial conducted in northern Taiwan. Nursing home residents (n = 43) were randomly assigned to either the control group or the experimental group. The control group received no extra care from the researcher while the experimental group received an individualised intervention and an eight-week follow-up. Participants were assessed using the Bristol Stool Form Scale, the Patient Assessment of Constipation Symptoms, types and dosages of laxative, and bowel sound observations. Data were taken at baseline, four weeks as well as eight weeks after the intervention. The participants in the experimental group had a significantly higher increase in the frequency of defecation (group effect, p = 0·029) and in bowel sounds (interaction effect, p = 0·010) compared to those in the control group. However, the two groups did not differ significantly in symptoms and the severity of the constipation symptoms, Bristol Stool Form and use of laxatives. The results of this trial suggest that the individualised intervention may be appropriate for decreasing constipation among nursing home residents and encourage further study and confirmation. Using individualised intervention to enhance the self-care ability related to constipation among older adults is recommended.


Measuring Use of Evidence-Based Psychotherapy for PTSD in VA Residential Treatment Settings with Clinician Survey and Electronic Medical Record Templates.
Available studies on implementation of evidence-based psychotherapy (EBP) for patients attending Department of Veterans Affairs (VA) residential post-traumatic stress disorder (PTSD) programs rely on therapist self-report of EBP delivery. Patient-level data on receipt of EBP are needed both to corroborate therapist self-report and to understand patient factors that predict receipt of EBPs for PTSD. We identified 159 therapists from 38 VA residential PTSD programs who responded to a survey about EBP implementation during the 2015 fiscal year (FY15). Therapists self-reported their use of two EBPs, including prolonged exposure delivered in an individual format (PE-I) and cognitive processing therapy delivered in individual and group formats (CPT-I and CPT-G). Using electronic medical record (EMR) templates mandated for EBP documentation in FY15, we measured contemporaneous patient-level receipt of EBPs for PTSD. We assessed the degree of correlation between therapist self-reported EBP delivery and patient receipt of EBT as measured by EMR templates using polychoric correlation coefficients. We determined patient and therapist factors that predicted the receipt of EBPs with multivariable logistic regression, using random effects and robust standard error estimation, and controlling for site. The Veterans IRB of Northern New England provided a waiver of informed consent; as this was a retrospective review, no patients or therapists were contacted, and all data were stored, transmitted, and analyzed on secure VA servers. The VA Connecticut Health Care System Human Research Protection Program approved secondary use of therapist survey data for this project. When EMR template use became mandated in FY15, the proportion of patients in residential PTSD programs who received at least one EBP session that was recorded with an EMR template increased dramatically from 8.8% to 33.9%. There was adequate correlation and between survey-based and EMR-based measures of EBP receipt, with polychoric correlation values of 0.77 for PE-I, 0.69 for CPT-I, and 0.82 for CPT-G. Multiple patient factors were positive (e.g., female gender) and negative (e.g., depressive disorders) predictors of receipt of EBPs, even after controlling for site. Among therapist factors, only EBP consultant or trainer status was a positive predictor of EBP provision and only therapist race was a negative predictor of EBT provision after controlling for site. Following a FY15 mandate, EMR templates documenting EBP delivery were widely used by therapists working in VA residential PTSD programs. EBP receipt measured using EMR templates was consistent with therapist self-report of EBT delivery. There were several patient-level predictors of EBP receipt and therapist-level predictors of EBP delivery. However, therapists most likely to deliver EBPs were clustered at a limited number of sites.


Target-site EPSPS Pro-106-Ser mutation in Conyza canadensis biotypes with extreme resistance to glyphosate in Ohio and Iowa, USA.
Documenting the diversity of mechanisms for herbicide resistance in agricultural weeds is helpful for understanding evolutionary processes that contribute to weed management problems. More than 40 species have evolved resistance to glyphosate, and at least 13 species have a target-site mutation at position 106 of EPSPS. In horseweed (Conyza canadensis), this p106 mutation has only been reported in Canada. Here, we sampled seeds from one plant (= biotype) at 24 sites in Ohio and 20 in Iowa, screened these biotypes for levels of resistance, and sequenced their DNA to detect the p106 mutation. Resistance categories were based on 80% survival at five glyphosate doses: S (0×), R1 (1×), R2 (8×), R3 (20×), or R4 (40×). The p106 mutation was not found in the19 biotypes scored as S, R1, or R2, while all 25 biotypes scored as R3 or R4 had the same proline-to-serine substitution at p106. These findings represent the first documented case of target-site mediated glyphosate resistance in horseweed in the United States, and the first to show that this mutation was associated with very strong resistance. We hypothesize that the p106 mutation has occurred multiple times in horseweed and may be spreading rapidly, further complicating weed management efforts.


A Fusarium graminearum xylanase expressed during wheat infection is a necrotizing factor but is not essential for virulence.
Fusarium graminearum is the fungal pathogen mainly responsible for Fusarium head blight (FHB) of cereal crops, which attacks wheat spikes, reducing crop production and quality of grain by producing trichothecene mycotoxins. Several cytohistological studies showed that spike infection is associated with the production of cell wall degrading enzymes. Wheat tissue, as in other commelinoid monocot plants, is particularly rich in xylan which can be hydrolyzed by fungal endo-1,4-β-xylanase. The FG_03624 is one of the most expressed xylanase genes in wheat spikes 3 days after inoculation and was heterologously expressed in the yeast Pichia pastoris. The recombinant protein (22.7 kDa) possessed xylanase activity and induced cell death and hydrogen peroxide accumulation in wheat leaves infiltrated with 10 ng/μl or in wheat lemma surface treated with 20 ng/μl. This effect reflects that observed with other described fungal xylanases (from Trichoderma reesei, Trichoderma viride and Botrytis cinerea) with which the FG_03624 protein shares a stretch of amino acids reported as essential for elicitation of necrotic responses. Several F. graminearum mutants with the FG_03624 gene disrupted were obtained, and showed about 40% reduction of xylanase activity in comparison to the wild type when grown in culture with xylan as carbon source. However, they were fully virulent when assayed by single floret inoculation on wheat cvs. Bobwhite and Nandu. This is the first report of a xylanase able to induce hypersensitive-like symptoms on a monocot plant.


Field-scale rice yield estimation based on UAV-based MiniSAR data with Ku band and modified water-cloud model of panicle layer at panicle stage.
Scientific and accurate estimation of rice yield is of great significance to food security protection and agricultural economic development. Due to the weak penetration of high frequency microwave band, most of the backscattering comes from the rice canopy, and the backscattering coefficient is highly correlated with panicle weight, which provides a basis for inversion of wet biomass of rice ear. To solve the problem of rice yield estimation at the field scale, based on the traditional water cloud model, a modified water-cloud model based on panicle layer and the radar data with Ku band was constructed to estimate rice yield at panicle stage. The wet weight of rice ear scattering model and grain number per rice ear scattering model were constructed at field scale for rice yield estimation. In this paper, the functional area of grain production in Xiashe Village, Xin'an Town, Deqing County, Zhejiang Province, China was taken as the study area. For the first time, the MiniSAR radar system carried by DJI M600 UAV was used in September 2019 to obtain the SAR data with Ku band under polarization HH of the study area as the data source. Then the rice yield was estimated by using the newly constructed modified water-cloud model based on panicle layer. The field investigation was carried out simultaneously for verification. The study results show: the accuracies of the inversion results of wet weight of rice ear scattering model and grain number per rice ear scattering model in parcel B were 95.03% and 94.15%; and the accuracies of wet weight of rice ear scattering model and grain number per rice ear scattering model in parcel C+D+E were over 91.8%. In addition, different growth stages had effects on yield estimation accuracy. For rice at fully mature, the yield estimation accuracies of wet weight of ear and grain number per ear were basically similar, both exceeding 94%. For rice at grouting stage, the yield estimation accuracy of wet weight of ear was 92.7%, better than that of grain number per ear. It was proved that it can effectively estimate rice yield using the modified water-cloud model based on panicle layer constructed in this paper at panicle stage at field scale.


Effects of 2-substituted 3-dimethylamino-5,6-methylenedioxyindenes on calcium-induced arrhythmias.
Pharmacological, biochemical, and electrophysiological studies have demonstrated that the 2-n-propyl (Pr) and 2-n-buty (Bu) 3-dimethylamino-5,6-methylenedioxyindenes (MDIs) are intracellular calcium antagonists. The MDIs exhibit coronary dilating properties in the in vitro rabbit heart and possess antiarrhythmic activity in vivo in the anesthetized ouabain-toxic dog. The results of the present investigation demonstrate that the MDIs possess significant antiarrhythmic activity against calcium-induced arrhythmias in anesthetized rats and dogs. Pretreatment of dogs with an acute dose (30 mg/kg, i.v.) of either MDI significantly delayed both the onset of intravenous infusion of calcium (500 mg/kg/hr, CaCl2 . 2H2O). Pretreatment of rats with an acute dose (3.75 mg/kg, i.v.) of either MDI provided virtually complete protection against bradycardia, arrhythmias, and death induced by an acute intravenous dose of calcium (1 ml/kg of a 10% CaCl2 . 2H2O solution. The antiarrhythmic potency of the MDIs in rats was equivalent to that of verapamil, but the latter drug produced bradycardia, electrocardiographic alterations, and atrioventricular block, whereas the MDIs exhibited no intrinsic deleterious effects of cardiac function. Prenylamine (a membrane calcium antagonist) and diphenylhydantoin demonstrated no antiarrhythmic properties in this rat model at comparable doses to the MDIs.


Transformation of Bacillus subtilis by single-stranded plasmid DNA.
The single-stranded form of a pE194-based plasmid transformed Bacillus subtilis protoplasts at least as efficiently as did the double-stranded plasmid, but the single-stranded form did not detectably transform B. subtilis competent cells.


Improving Oxygen Reduction Performance by Using Protic Poly(Ionic Liquid) as Proton Conductors.
Improving catalytic performance of the oxygen reduction reaction (ORR) of Pt/C catalysts is essential for reducing Pt-loading and the according cost of proton exchange membrane fuel cells (PEMFCs). Herein, we report a new conceptual design of catalyst layers to improve the ORR performance of Pt/C catalysts by replacing perfluorosulfonated ionomers with protic poly(ionic liquid) as a proton conductor. The specific activity of the designed catalyst at 0.9 V under acidic conditions is over three times higher than that of catalyst using Nafion as the proton conductor. Furthermore, the durability test reveals that the introduction of protic poly(ionic liquid) ionomers can protect Pt nanoparticles against aggregation during potential cycles, but it is less durable than Nafion because of the nature of hydrocarbons. Nevertheless, we believe that replacing perfluorosulfonated ionomers with protic poly(ionic liquid) as proton conductors could be a promising strategy to design an efficient cathode for low Pt-loading PEMFCs.


In vitro antiplasmodial activity and chloroquine-potentiating action of three new isoquinoline alkaloid dimers isolated from Hernandia voyronii Jumelle.
Three new isoquinoline alkaloid dimers with pavine and benzyl-tetrahydroisoquinoline moieties have been isolated, by bioassay-guided fractionation, from an alkaloid extract of Hernandia (= Hazomalania) voyronii Jumelle. All had inherent antiplasmodial activity in vitro and two enhanced chloroquine effectiveness in vitro in a dose-dependent manner. Some structure requirements for the chloroquine-potentiating action were identified in the isoquinoline series. These results, if confirmed in vivo, would account for the traditional use of H. voyronii as an adjuvant to complement chloroquine action in Malagasy herbal remedies.