MicroRNA-325 targets lipocalin 15 to suppress proliferation, migration and invasion of breast cancer cells.
Recent studies have proved the diverse roles of miRs in different cancer-related processes. This study was undertaken to determine the therapeutic implications of miR-325-3p in breast cancer. Expression analysis was carried out by qRT-PCR. Transfections were performed by Lipofectamine 2000 reagent. MTT assay was used for cell viability. Transwell assays were used for cell migration and invasion. Western blot analysis was used for protein expression analysis. Gene expression analysis revealed miR-325 to be significantly suppressed in breast cancer tissues and cell lines. Nonetheless, ectopic expression of miR-325 resulted in suppression of the growth and colony development potential of the SK-BR-3 and CAMA-1 cells. Transwell assays showed that miR-325 overexpression also resulted in the decline of the migration and invasion of the SK-BR-3 and CAMA-1 cells. Bioinformatic analysis showed that miR-325 targets lipocalin 15 (LNC15) in breast cancer cells. LNC15 was also overexpressed in the breast cancer tissues and cell lines. However, overexpression of miR-325 caused a significant decline in the LNC15 expression in SK-BR-3 cells. Additionally, silencing of LNC15 resulted in inhibition of the growth, migration and invasion of the SK-BR-3 cells. Rescue assay showed that overexpression of LNC15 could promote the growth, migration and invasion of the miR-325 overexpressing effects. Taken together, the evidence shows that miR-325 acts as a tumor suppressor in breast cancer and may be used in the treatment of breast cancer.


[Cytophotometry of myelokaryocyte DNA following a single exposure to low-intensity UHF irradiation].
DNA content of nondifferentiated and blast bone marrow cells was studied cytophotometrically after single exposure to SHF-radiation. It is supposed that cell adaptive reactions in blood system are responsible for phase changes in DNA quantity, the passage of cells through the mitotic cycle and their proliferative activity.


The response of different kinds of rapeseed cultivars to foliar application of nitrogen, sulphur and boron.
The study was based on in 2016-2017, 2017-2018, 2018-2019 field experiment conducted at the University of Natural Sciences and Humanities in Siedlce (Zawady Agricultural Experimental Station), eastern Poland. The studied factors were: I. winter rape cultivars: Monolit-open pollinated cultivar; PT248-F1 hybrid cultivars with traditional growth type; PX115-F1 hybrid cultivars with a semi-dwarf growth type and types of foliar nutrition: (1) control variant without foliar nutrition and amino acids; (2) amino acid; (3) foliar fertilizer sulphur and boron; (4) foliar fertilizer sulphur with foliar fertilizer boron and amino acid. The aim of the study was to determine the effect of foliar application of sulphur, boron, amino acids on the use and feed value of seeds of three winter rape morphotypes. The highest content of crude fat in seeds of the studied morphotypes was found after foliar fertilization with sulphur and boron and amino acids, while the lowest under the influence of amino acids. The highest concentration of total protein was obtained after the application of amino acids, and regardless of the morphotype studied on this object, the same value of this trait was demonstrated. In restored morphotypes, the use of additional foliar fertilization S and B in combination with amino acids did not significantly increase this characteristic compared to the amino acid variant. As a result of the application of amino acids and foliar feeding of S and B, and amino acids, the largest increase in crude fibre content in rapeseeds was obtained, while the application of S and B did not significantly increase this characteristic compared to the control variant. The best feed and use value of seeds were noted in restored morphotypes, with the semi-dwarf variety having the highest fat and crude fibre content. Climatic conditions in the years of research significantly determined the value of seeds. The highest values of the examined traits were obtained in the last year of the study, and the lowest in the growing season 2016-2017.


Surface-tension-induced double emulsion drops via phase separation of polymeric fluid confined in micromolds for capsule templates.
We report a simple and rapid route to produce double emulsion drops by utilizing phase separation of the confined fluid in micromolds and surface-tension-induced drop formation. Specifically, we use cross-shaped micromolds containing prepolymer solution that phase-separates into two compartments upon addition of wetting fluid with separation agent (SA). Subsequently, Laplace pressure-driven flow allows it to form double emulsion drops without use of any surfactants and complex formulations of fluids. The size of each compartment in the emulsion drops can be controlled by tuning composition of the prepolymer solution and separation agent, making the double emulsion drops with varying shell thicknesses. The phase separation creates two compartments with different polarity (i.e. water-soluble and water-insoluble), enabling encapsulation of both hydrophilic and/-or hydrophobic cargoes in desired compartments depending on their solubility. In addition, we produce poly(N-isopropylacrylamide) (pNIPAm) hydrogel microcapsules by solidifying middle phase in the double emulsion drops; thus, hydrophilic large cargo loaded priorly in the core can be encapsulated within hydrogel shells. Finally, by taking advantage of hydrophilic-hydrophobic phase transition behavior of pNIPAm, we achieve encapsulation of small cargo via post-loading approach; the encapsulated cargo can be released by tuning temperature.


Pediatric reference intervals for lymphocyte vitamin C (ascorbic acid).
To establish pediatric reference intervals for lymphocyte vitamin C. This was a prospective study of 194 well children aged 0-7 years old of mixed ethnicity who had blood drawn for the purpose of this study. Blood was collected during elective surgery under general anesthesia and lymphocytes isolated and stored as frozen ascorbic acid lymphocyte lysates for later HPLC analysis by previously described methodology. Reference intervals were established according to the Clinical and Laboratory Standards Institute (CLSI) and the International Federation of Clinical Chemistry (IFCC) guidelines (C28-A3). Horn-Pesce robust method was used to estimate the 95% confidence interval and 95% reference interval. Reference intervals were independent of age or gender and shown to be 12.9-52.8 μg/10(8) cells (lymphocytes). We have defined pediatric reference ranges for lymphocyte vitamin C in healthy, fasted children at a relevant age group (0-7 years). The new reference interval can now be used to more reliably explore possible implications of variation of vitamin C levels on bleeding and other clinical signs.


Functional FCGR3A 158 V/F and IL-6 -174 C/G polymorphisms predict response to biologic therapy in patients with rheumatoid arthritis: a meta-analysis.
The aim of this study was to investigate whether the Fc gamma receptor 3A (FCGR3A) 158 V/F and interleukin-6 (IL-6) promoter -174 G/C polymorphisms can predict the response to biologic-based therapy in patients with rheumatoid arthritis (RA). We conducted a meta-analysis of studies on the association between the FCGR3A V/F polymorphism or the IL-6 -174 C/G polymorphism and non-responsiveness to biologic therapy in RA patients. A total of 10 studies involving 1,427 patients were considered. These studies consisted of seven studies on the FCGR3A polymorphism and three studies on the IL-6 polymorphism. Meta-analysis showed no association between the FCGR3A VV+VF genotype and non-responders to biologic therapy [odds ratio (OR) 0.881, 95 % confidence interval (CI) 0.505-1.537, p = 0.655]. However, stratification by biologic type indicated an association between the FCGR3A VV+VF genotype and non-responders to rituximab (OR 0.566, 95 % CI 0.373-0.857, p = 0.007), but no association was found in non-responders to tumor necrosis factor (TNF)-blockers (OR 1.337, 95 % CI 0.869-2.056, p = 0.186). Meta-analysis revealed no association between the IL-6 CC+CG genotype and non-responders to the biologics (OR 3.233, 95 % CI 0.766-13.64, p = 0.110). However, an association was found between the IL-6 CC+CG genotype and non-responders to anti-TNF therapy (OR 8.030, 95 % CI 1.807-33.68, p = 0.006). This meta-analysis demonstrates that FCGR3A V allele carriers show a better response to rituximab, and individuals carrying the IL-6 -174 C allele show a poorer response to anti-TNF therapy for RA. Genotyping for these polymorphisms may be a useful tool for predicting the response to biologics with respect to personalized medicine.


Carnosine attenuates renal ischemia-reperfusion injury by inhibiting GPX4-mediated ferroptosis.
Increasing evidence and our preliminary work have revealed the significant role of ferroptosis in acute kidney injury (AKI) induced by ischemia/reperfusion (IR). Carnosine (Car), a dipeptide consisting of β-alanine and L-histidine, has been shown to ameliorate HG-induced tubular epithelial cells inflammation. Whether Car exerts protective effects on AKI, and its molecular mechanism have not been clarified. Our in vivo and in vitro IR-AKI mouse models demonstrated that Car alleviates kidney injury, inflammation and ferroptosis. In hypoxia/reoxygenation (HR) induced human renal tubular epithelial cells (HK2), Car treatment reduced lipid peroxidation and iron accumulation, suppressed oxidative stress, and inhibited ferroptosis. Through cellular thermal shift assay (CETSA) and molecular docking, we identified GPX4 as a potential target that binds with Car. Further study showed that overexpressed GPX4 had a comparable protective effect on HK2 cells under HR conditions, similar to Car. Additionally, our findings demonstrated that Car exhibited similar anti-ferroptosis effects in both folic acid (FA)-induced AKI mouse models and Erastin induced HK2 cells. In conclusion, our results highlight that Car alleviate renal IR injury by inhibiting GPX4-mediated ferroptosis. Car shows promise as a potential therapeutic drug for IR-AKI and other diseases associated with ferroptosis.


Immunosensor with a controlled orientation of antibodies by using NeutrAvidin-protein A complex at immunoaffinity layer.
The orientation of antibody was controlled by using NeutrAvidin-protein A complex on the gold surface of SPR biosensor. The surface density of receptor antibody (anti-hIgG) was compared by treatment of receptor antibody to the layer of avidin, NeutrAvidin, protein A, NeutrAvidin-protein A complex and bare gold surface of SPR biosensor. The ligand antibody (hIgG) was injected to each IA layer and the binding ratio of ligand antibody per unit receptor was estimated as a parameter of orientation control. The NeutrAvidin-protein A complex on gold surface of SPR biosensor showed the highest surface density of receptor antibody as well as the binding ratio of ligand antibody per receptor antibody. The NeutrAvidin-protein A complex was also prepared on biotin-labelled SAM, and the binding ratio of ligand per receptor was found to be significantly improved in comparison to the IA layer prepared by chemical coupling of receptor antibody to the SAM layer. The NeutrAvidin-protein A complex which showed the highest efficiency for the binding of ligand antibodies, was applied for the detection of a cancer marker called CEA. By using NeutrAvidin-protein A complex and sandwich assay for signal amplification, sensitivity was improved to be 1.5-fold higher than bare gold surface and the detection of CEA with the detection limit of 30 ng/ml was achieved.


Synergistic Activity of Fluoroquinolones Combining with Artesunate Against Multidrug-Resistant Escherichia coli.
Multidrug resistance (MDR) is an increasing public health concern worldwide. Artesunate (ART) has been reported to be significantly effective in enhancing the effectiveness of various β-lactam antibiotics against MDR Escherichia coli via inhibiting the efflux pump genes. Apart from β-lactam antibiotics, there is no report regarding the potential synergistic effects of ART combining with fluoroquinolones (FQs). In this study, we investigated whether ART can enhance the antibacterial effects of FQs in vitro. The antibacterial activity of ART and antibiotics against 13 animal-derived E. coli clinical isolates was assessed for screening MDR strains. Then the synergistic activity of FQs with ART against MDR E. coli isolates was evaluated. Daunorubicin (DNR) accumulation within E. coli and messenger RNA (mRNA) expressions of acrA, acrB, tolC, and qnr genes were investigated. The results showed that ART did not show significant antimicrobial activity. However, a dramatically synergistic activity of ART combining with FQs was obsessed with (ΣFIC) = 0.12-0.33. ART increased the DNR accumulation and reduced acrAB-tolC mRNA expression, but enhanced the mRNA expression of qnrS and qnrB within MDR E. coli isolates. These findings suggest that ART can potentiate FQs activity which may be associated with drug accumulation by inhibiting the expression of acrAB-tolC.


[Diagnosis of yeast contamination in urine especially of patients with indwelling catheters (authors transl)].
Mycologic investigations of female patients with permanent catheters have shown that the yeast contamination detected in the urine depends on the time of catheterizing. Yeast from the genital and anal regions are the main sources of infection. Prophylactic therapy with antimycotic agents is recommended.


Upregulation of Glypican-3 expression in hepatocellular carcinoma but downregulation in cholangiocarcinoma indicates its differential diagnosis value in primary liver cancers.
Expression alteration of Glypican-3 (GPC3) is associated with several malignancies and has been identified as an overexpressed gene in hepatocellular carcinoma (HCC). In this study GPC3 expression in intrahepatic chanlangiocarcinoma (ICC), gallbladder cancer and HCC was quantitatively detected. Real-time quantitative reverse transcription polymerase chain reaction was used to detect the expression level of GPC3. GPC3 expression was elevated more than two-fold in HCC compared with adjacent tissue in 90 of 100 HCC cases. The average expression level of GPC3 was significantly higher in HCC than that in adjacent liver tissues (P<0.0001). Only in four of 21 ICC cases GPC3 expression was upregulated more than two-fold in tumor tissues. GPC3 expression was downregulated in gallbladder cancer in 12 of 13 cases and the average expression level was significantly lower than that in normal gallbladder tissues (P<0.05). The different expression patterns of GPC3 in HCC and ICC suggested that it might play a different role in theses tumors and could serve as a biomarker for differential diagnosis of HCC and ICC.


[Diffuse neoplastic infiltration of leptomeninges by malignant pleural mesothelioma].
A 65 year-old-man, diagnosed with malignant pleural mesothelioma, developed polymorphic neurologic symptoms and signs indicating involvement of cranial and spinal nerves. Necropsy disclosed a diffuse neoplastic invasion of the leptomeninges with no other neural metastases.


Setteleia, a New Genus for Four New Species from the Philippines (Insecta: Lepidoptera: Erebidae: Arctiinae).
We describe a new Lithosiini genus Setteleia gen. nov. for four new species from the Philippines: S. witti sp. nov. (Mindanao Island), S. carota sp. nov. (northern Luzon Island), S. lourensi sp. nov. (eastern Luzon Island) and S. bakunawa sp. nov. (northern Luzon Island). The new genus belongs to the Asura Walker, 1854 /Miltochrista Hübner, [1819] generic complex and is related to the genera Moorasura Volynkin & Huang, 2019, Sarbine Volynkin, 2019, Ammatho Walker, 1855 and Cyme Felder, 1861 and also shares some genital characters with such genera as Integrivalvia Volynkin & Huang, 2019, Fossia Volynkin, Ivanova & Huang, 2019 and Asuridia Hampson, 1900, but is characterized by a number of autapomorphic features in both male and female genitalia.


Toxicological effects and reproductive impairments in female perch (Perca fluviatilis) exposed to leachate from Swedish refuse dumps.
We have previously found that leachate from a Swedish refuse dump caused toxicological effects, including endocrine disruption and reproductive failures, in feral female perch (Perca fluviatilis) from Molnbyggen and in brook trout (Salvelinus fontinalis) from Vadbäcken. This raised concerns that leachate-induced toxicity might affect fish in other leachate-contaminated lakes. This study looks at female perch from three different regions in Sweden, focusing on toxicological biomarkers (skin lesions, liver-somatic index (LSI), ethoxyresorufin O-deethylase (EROD) activity and DNA adducts) and reproductive biomarkers (number of mature females, gonadosomatic index (GSI), aromatase (P450arom), progesterone, 17alpha-hydroxyprogesterone (17alpha-OHP), testosterone (T) and 17beta-estradiol (E2)). Five out of seven contaminated locations had lower numbers of mature females and most of them exhibited higher frequencies of fin erosion relative to their comparable reference sites. Females from Lake Nedre Vättern exhibited these effects, as well as body sores, high EROD activity, high levels of DNA adducts in the liver and the intestinal mucosa, low GSI and low plasma levels of T and E2, similar to the anti-estrogenic effects earlier found in SM perch from Molnbyggen in 1997 and in brook trout from Vadbäcken. No effects on LSI, GSI, aromatase, or circulating steroids were found in mature females from Molnbyggen in this study. This indicated less leachate-contamination, but low numbers of mature females in Lake Siljan, at the sewage treatment plant which now processes the leachate that earlier contaminated Molnbyggen and Vadbäcken, suggested insufficient sewage treatment at this site. All females in Lake Simshyttsjön were mature, and their high GSI and LSI, low EROD activity, high level of 17alpha-OHP and low level of T, suggest an estrogenic rather than an anti-estrogenic effect. The results show that our earlier findings in Molnbyggen and Vadbäcken are not common, but neither are they unique. This study also shows that refuse dumps should be considered as potential point-sources for environmental pollutants, and that uncontrolled leachate-contamination of lakes and freshwater reservoirs could be a serious environmental hazard for both wildlife and humans.


Olfactory cleft disease: an analysis of 13 cases.
Olfactory cleft disease was defined as (1) an olfactory disability related with a clinical and/or radiologic abnormality of the olfactory cleft and (2) a pathologic process limited to or predominating in the olfactory cleft. The objectives of this study were to describe the clinical and radiologic findings of this entity and to report the effect of medical therapy on the sense of smell. Thirteen patients with the criteria of olfactory cleft disease were included in this retrospective study. All patients had bilateral olfactory cleft abnormalities. A complete ENT examination with endoscopic evaluation and a computed tomography scan were performed in all cases. An olfactory threshold test was available in 12 cases before and after medical therapy, which included oral and topically administered steroid therapy. Olfactory disability was the major symptom of olfactory cleft disease. Other symptoms of chronic rhinosinus dysfunction was present in 75% of cases. Three pathologic processes were identified in patients with olfactory cleft disease: malformative, inflammatory, and inflammatory associated with anatomical deformities of olfactory cleft boundaries. Medical therapy was effective in lowering olfactory thresholds in 25% of the cases. Olfactory cleft disease involves various pathophysiologic processes. Computed tomography scanning provides the most information for diagnosis. Indications of functional endoscopic surgery remain to be defined after failure of medical therapy.


Strontium ranelate prevents bone loss in a rat model of localized muscle paralysis.
Twenty-one 3.5-month-old female Sprague-Dawley rats were randomly assigned to three groups: BTX group, in which each rat received a single intramuscular injection of 2 U of Clostridium botulinum toxin (BTX) in the quadriceps femoris muscle of the right hind limb; BTX + SR group, in which each rat received a BTX injection and a dose of strontium ranelate (dose level of 625 mg/kg/day); and the control group. All the rats were killed at 9 weeks post-treatment. It was showed that BTX-induced rats a rapid loss of body weight in the first 3 weeks, after which their body weight showed a slow increase similar to that observed in the control rats. The net body weight loss was mainly attributed to muscle atrophy. BTX caused remarkable bone degradation in either the trabecular bone or the cortical bone of the disuse femur. The deteriorations in the bone mass and bone microstructure were locally limited and could be prevented by strontium ranelate treatment. Biomechanical analysis showed that strontium ranelate treatment improved the mechanical performance of the tibia in BTX-treated rats. It was showed that a clinical-corresponding dose of strontium ranelate could prevent bone loss in long-term immobilized rats.


Amentoflavone, a plant biflavone: a new potential anti-inflammatory agent.
Biflavonoid is one of unique classes of naturally-occurring bioflavonoids. Certain biflavonoids including amentoflavone were previously reported to have inhibitory effect on the group II phospholipase A2 activity. Amentoflavone was also found to inhibit cyclooxygenase from guinea-pig epidermis without affecting lipoxygenase. In this study, anti-inflammatory and analgesic activities of amentoflavone were evaluated. When amentoflavone was administered intraperitoneally, it showed a potent anti-inflammatory activity as determined by amelioration of croton-oil induced mouse ear edema. It also showed a potent anti-inflammatory activity in the rat carrageenan paw edema model (ED50 = 42 mg/kg) compared to the activity of prednisolone (35 mg/kg) and indomethacin (10 mg/kg). However, amentoflavone did not show a significant inhibitory activity against rat adjuvant-induced arthritis, a chronic inflammatory model. In addition, amentoflavone was found to possess a potent analgesic activity in the acetic acid writhing test (ED50 = 9.6 mg/kg) compared to the activity of indomethacin (3.8 mg/kg). These results suggest that amentoflavone may be a potential lead for a new type of anti-inflammatory agents having dual inhibitory activity of group II phospholipase A2 and cyclooxygenase.


[Temporary epicardial pacing following cardiac surgery: practical aspects].
To review the practical aspects of temporary epicardial pacing following open heart surgery. Review of articles published in English or French within the last five years and investigating temporary epicardial pacing (indications, pacing modes, epicardial wires and temporary generators). The studies were extracted from the databases ScienceDirect and Pubmed. Temporary epicardial pacing is used to treat severe conduction and/or rhythm disorders, but also to improve hemodynamics by optimizing selected temporary pacing settings. Temporary epicardial pacing consists in choosing the most suitable pacing mode according to the situation (surgery, patient, conduction and/or rhythm abnormalities) and setting its parameters that ensure : i) optimal pacemaker functioning; ii) epicardial electrodes longevity; iii) the most favorable hemodynamic profiles. Management of temporary pacing settings and their regular adjustment, at least daily and based on thresholds, are part of good clinical practices. Nevertheless, the French literature lacks official recommendations for temporary epicardial pacing. Temporary epicardial pacing following cardiac surgery is a simple method, more effective than transcutaneous pacing and easier to implement than transvenous pacing. Its practical management should be known by all physicians (anesthetists, cardiac surgeons) as well as paramedical personnel in order to avoid the risks of suboptimal functioning. A good practice protocol is proposed at the end of the manuscript.


[Isolation of the RNA-poly A+ fraction from mitochondria of Drosophila melanogaster embryos. Incorporation of methionine-35S into newly-synthesized proteins in whole and lysed mitochondria].
The poly A+ RNA fraction of embryos mitochondria of Drosophila melanogaster was prepared by fractionation by oligo dT-cellulose chromatography. It accounted for 5% of total insoluble RNA. 15 bands were identified by electrophoresis, from 500 to 2,000 base pairs. This poly A+ RNA fraction hybridizes with mitochondrial DNA clones. It contains mitochondrial transcripts: messenger RNA and the large ribosomal RNA. The sum of the transcripts (29 kb) exceed the coding capacities of the mitochondrial DNA. Some of the transcripts may be precursors, greater in size, of the mRNA corresponding to a gene. Our results are compared to those previously published (9, 11, 12). The relationship between mitochondrial transcripts and proteins was studied in whole and lysed isolated mitochondria. In the former, incubated in the presence of 35S-methionine, 20 to 30 radioactive bands were identified (M. Wt. 100 to 16 kd). This incorporation was totally inhibited by chloramphenicol. When the mitochondria were opened in the presence of detergent (lysis). 35S-methionine incorporation was markedly enhanced. In all cases the number of proteins identified was greater than the number of potential genes (12). This finding is discussed.


Understanding the role of magnesium stearate in lowering punch sticking propensity of drugs during compression.
The sticking of active pharmaceutical ingredient (API) to the surfaces of compaction tooling, frequently referred to as punch sticking, causes costly downtime or product failures in commercial tablet manufacturing. Magnesium stearate (MgSt) is a common tablet lubricant known to ameliorate the sticking problem, even though there exist exceptions. The mechanism by which MgSt lowers punch sticking propensity (PSP) by covering API surface is sensible but not yet experimentally proven. This work was aimed at elucidating the link between PSP and surface area coverage (SAC) of tablets by MgSt, in relation to some key formulation properties and process parameters, namely MgSt concentration, API loading, API particle size, and mixing conditions. The study was conducted using two model APIs with known high PSPs, tafamidis (TAF) and ertugliflozin-pyroglutamic acid (ERT). Results showed that PSP decreases exponentially with increasing SAC by MgSt. The composition of material stuck to punch face was also explored to better understand the onset of punch sticking and the impact of possible MgSt-effected punch conditioning event.


MiR-507 inhibits the growth and invasion of trophoblasts by targeting CAMK4.
To elucidate the potential influences of miR-507 and CAMK4 on the progression of preeclampsia (PE). Placental tissues were collected from 24 PE pregnancies and 24 healthy pregnancies. The relative levels of miR-507 and CAMK4 in placental tissues were detected. In addition, expressions of apoptosis-associated genes in collected tissues were examined by both quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The influences of miR-507 and CAMK4 on proliferative and migratory abilities in HTR-8/SVneo cells were assessed by CCK-8 and transwell assay, respectively. The target relationship between miR-507 and CAMK4 was detected by Luciferase assay. MiR-507 was upregulated in placental tissues collected from PE pregnancies. Overexpression of miR-507 suppressed proliferative and migratory abilities, and stimulated apoptosis in HTR-8/SVneo cells. CAMK4 was the target gene of miR-507, which was downregulated in placental tissues collected from PE pregnancies and negatively correlated to miR-507 level. The knockdown of CAMK4 suppressed proliferative and migratory abilities, and stimulated apoptosis in HTR-8/SVneo cells, and these trends were abolished by silence of miR-507. Highly expressed miR-507 in PE pregnancies inhibits proliferative and migratory potentials, and induces apoptosis in trophoblasts by targeting CAMK4.


Effect of aortic bifurcation geometry on pressure and peak wall stress in abdominal aorta: Fluid-structure interaction study.
Geometry of aorto-iliac bifurcation may affect pressure and wall stress in aorta and thus potentially serve as a predictor of abdominal aortic aneurysm (AAA), similarly to hypertension. Effect of aorto-iliac bifurcation geometry was investigated via parametric analysis based on two-way weakly coupled fluid-structure interaction simulations. The arterial wall was modelled as isotropic hyperelastic monolayer, and non-Newtonian behaviour was introduced for the fluid. Realistic boundary conditions of the pulsatile blood flow were used on the basis of experiments in literature and their time shift was tailored to the pulse wave velocity in the model to obtain physiological wave shapes. Eighteen idealized and one patient-specific geometries of human aortic tree with common iliac and renal arteries were considered with different angles between abdominal aorta (AA) and both iliac arteries and different area ratios (AR) of iliac and aortic luminal cross sections. Peak wall stress (PWS) and systolic blood pressure (SBP) were insensitive to the aorto-iliac angles but sensitive to the AR: when AR decreased by 50%, the PWS and SBP increased by up to 18.4% and 18.8%, respectively. Lower AR (as a result of the iliac stenosis or aging), rather than the aorto-iliac angles increases the BP in the AA and may be thus a risk factor for the AAA development.


Physical activity and its association with plasma lipids and lipoproteins in elderly women.
The association of physical activity to plasma lipid and lipoprotein levels was examined cross-sectionally in 634 elderly, postmenopausal women whose mean age was 70.7 years. Leisure-time physical activity in kilocalories per week was assessed by the Paffenbarger Questionnaire, a composite index of sports/recreation, stair climbing, and walking. Subjects were categorized by tertile of kilocalories per week from total physical activity, number of flights of stairs climbed, and number of blocks walked. After adjustment for age, body mass index, education, and oral estrogen use, no significant differences were noted in total cholesterol, high-density-lipoprotein cholesterol (HDL-C), HDL2-C, HDL3-C, low-density-lipoprotein cholesterol (LDL-C), or triglycerides across tertile of total activity or number blocks walked. Total cholesterol was significantly different across tertile of number of flights of stairs climbed (P < 0.05), and only a borderline association was noted with LDL-C (P < 0.08). The study suggests that leisure-time physical activity, as measured by the Paffenbarger Questionnaire, is not associated with a favorable lipid profile in elderly, postmenopausal women. Failure to find an association may reflect the relatively small number of women who engaged in moderate- or high-intensity activity.


The decreased membrane fluidity of in vivo aged, human erythrocytes. A spin label study.
The decreased membrane fluidity of the in vivo aged, human erythrocytes is found, by monitoring the electron paramagnetic resonance (EPR) spectra of fatty acid spin labels incorporated into the membrane. In addition, the decreased cell sizes and the decreased cholesterol and phospholipids contents, without significant changes of the quantity of the membrane proteins, also the decrease of ATP and 2,3-diphosphoglycerate and the increase of ADP and AMP, in the aged cells, were observed. Further the functional impairments of the aged cells, i.e. the increased oxygen affinity and the decreased deformability, were shown. On the basis of these quantitative data, the alteration of the protein-lipid organization, due to decreased lipid/protein ratio, the modified protein-lipid interaction and/or the influences of the diminished ATP content, is suggested to contribute towards the decreased membrane fluidity of the in vivo aged erythrocytes.


A label-free liquid crystal-assisted aptasensor for trace level detection of tobramycin in milk and chicken egg samples.
Developed herein is an aptasensing array based on liquid crystal (LC) for monitoring of tobramycin (TOB) antibiotic. The direction of LC molecules from vertical to a random status was induced by the conformational changes of the specific aptamer due to its selective interaction with the target. The dark view of the aptasensing platform changed to colorful through observation by a polarized light microscope that clarifies the TOB presence. The aptasensor is especially able to determine TOB in the linear concentration range of 0.005-600 pM with a limit of detection (LOD) as 0.0021 pM. The TOB values can be determined successfully in the milk and chicken egg samples that highlights the potential applicability of the designed aptasensor. The proposed sensing approach is facile, operator-independent, label-free, and ultra-sensitive, making it novel for developing real-time portable sensing devices for future.


Non-reproducible sequence artifacts in FFPE tissue: an experience report.
Recent advances in sequencing technologies supported the development of molecularly targeted therapy in cancer patients. Thus, genomic analyses are becoming a routine part in clinical practice and accurate detection of actionable mutations is essential to assist diagnosis and therapy choice. However, this is often challenging due to major problems associated with DNA from formalin-fixed paraffin-embedded tissue which is usually the primary source for genetic testing. Here we want to share our experience regarding major problems associated with FFPE DNA used for PCR-based sequencing as illustrated by the mutational analysis of ERBB4 in melanoma. We want to focus on two major problems including extensive DNA fragmentation and hydrolytic deamination as source of non-reproducible sequence artifacts. Further, we provide potential explanations and possible strategies to minimize these difficulties and improve the detection of targetable mutations. Genomic DNA from formalin-fixed paraffin-embedded tumor samples was isolated followed by PCR amplification, Sanger sequencing and statistical analysis. Analysis of Sanger sequencing data revealed a total of 46 ERBB4 mutations in 27 of 96 samples including the identification of 11 mutations at three previously unknown mutational hotspots. Unfortunately, we were not able to confirm any assumed hotspot mutation within repeated sequencing of relevant amplicons suggesting the detection of sequence artifacts most likely caused by DNA lesions associated with FFPE tissues. Since DNA from FFPE tissue is usually the primary source for mutational analyses, appropriate measures must be implemented in the workflow to assess DNA damage in formalin-fixed tissue to ensure accurate detection of actionable mutations and minimize the occurrence of sequence artifacts.


Determination of phenotypes of phosphoglucomutase (PGM1) in bloodstains by cellulose acetate electrophoresis.
A good separation of PGM1 isoenzymes from bloodstains has been obtained with an adapted Sonneborn method of electrophoresis on cellulose acetate. This method requires very small quantities of the examined material, a short time of electrophoresis, and is cheaper and simpler than hitherto described. A correct determination of PGM1 phenotypes from bloodstains was easy up to 24 weeks. Occasionally it was possible to determine phenotypes in bloodstains as old as 47 weeks, and in one case even in bloodstains 126 weeks old. This method may be useful in the forensic serology for determination of PGM1 phenotypes from bloodstains.


Altered appetitive conditioning in overweight and obese women.
Overweight and obese individuals show increased psychological and physiological reactivity to food cues and many of them have difficulties in achieving long-term weight loss. The current study tests whether abnormalities in the learning and extinction of appetitive responses to food cues might be responsible for this. Overweight/obese and healthy weight women completed a differential appetitive conditioning task using food as rewards, while eating expectancies, eating desires, conditioned stimulus evaluations, salivation, and electrodermal responses were assessed during an acquisition and extinction phase. Results suggested reduced discriminative conditioning in the overweight/obese group, as reflected by a worse acquisition of differential eating desires and no successful acquisition of differential evaluative responses. Some evidence was also found for impaired contingency learning in overweight and obese individuals. No group differences in conditioned salivation and skin conductance responses were found and no compelling evidence for differences in extinction was found as well. In sum, the current findings indicate that overweight and obesity may be characterized by reduced appetitive conditioning. It is suggested that this could be causally related to overeating via stronger context conditioning or a tendency towards overgeneralization in overweight and obese individuals.


Modelling extreme risks for green bond and clean energy.
Value at Risk and Expected Shortfall are two traditional tools used to measure extreme risk in financial markets. However, there is little research on measuring extreme risk in emerging markets such as green bonds and clean energy. This paper uses both semi-parametric models with simultaneous excitation functions and traditional models to estimate extreme risk in SP500 Green Bond (GB) and Global Clean Energy (GCE), selecting Expected Shortfall (ES) and Value at Risk (VaR) as the indices of extreme risk. Then, the paper uses a breakpoint scan of the predictions of the different types of models. The results find that the green bond market was relatively stable while the global clean energy market experienced sharp fluctuations after the COVID-19 pandemic outbreak. Representative models in GCE have at least two breakpoints, but those for GB have no breakpoints. The GARCH model with normal innovations performs the best among all target models, and the GARCH-FZ model fits the best among all semi-parametric candidates. Our research could help governments, companies, and investors with risk management and improve model accuracy and mechanisms for measuring extreme risks.


Collection and Analysis of Multimodal Data for SUDEP Biomarker Discovery.
This paper discusses the acquisition and processing of multimodal physiological data from patients with epilepsy in Epilepsy Monitoring Units for the discovery of risk factors for Sudden Expected Death in Epilepsy (SUDEP) that can be combined through integrative analysis for biomarker discovery.


Baseline clusters and the response to positive airway pressure treatment in obstructive sleep apnoea patients: longitudinal data from the European Sleep Apnea Database cohort.
The European Sleep Apnea Database was used to identify distinguishable obstructive sleep apnoea (OSA) phenotypes and to investigate the clinical outcome during positive airway pressure (PAP) treatment. Prospective OSA patient data were recruited from 35 sleep clinics in 21 European countries. Unsupervised cluster analysis (anthropometrics, clinical variables) was performed in a random sample (n=5000). Subsequently, all patients were assigned to the clusters using a conditional inference tree classifier. Responses to PAP treatment change in apnoea severity and Epworth sleepiness scale (ESS) were assessed in relation to baseline patient clusters and at short- and long-term follow-up. At baseline, 20 164 patients were assigned (mean age 54.1±12.2 years, 73% male, median apnoea-hypopnoea index (AHI) 27.3 (interquartile range (IQR) 14.1-49.3) events·h-1, and ESS 9.8±5.3) to seven distinct clusters based on anthropometrics, comorbidities and symptoms. At PAP follow-up (median 210 [IQR 134-465] days), the observed AHI reduction (n=1075) was similar, whereas the ESS response (n=3938) varied: largest reduction in cluster 3 (young healthy symptomatic males) and 6 (symptomatic males with psychiatric disorders, -5.0 and -5.1 units, respectively (all p<0.01), limited reduction in clusters 2 (obese males with systemic hypertension) and 5 (elderly multimorbid obese males, -4.2 (p<0.05) and -3.7 (p<0.001), respectively). Residual sleepiness in cluster 5 was particularly evident at long-term follow-up (p<0.05). OSA patients can be classified into clusters based on clinically identifiable features. Importantly, these clusters may be useful for prediction of both short- and long-term responses to PAP intervention.


Polyisoprenoid amphiphilic compounds as inhibitors of squalene synthesis and other microsomal enzymes.
Analogues of farnesyl pyrophosphate containing a farnesyl moiety and a variety of amine residues replacing the pyrophosphate have been synthesized. Most of these compounds were effective inhibitors of the synthesis of squalene and presqualene pyrophosphate from farnesyl pyrophosphate. 50% inhibition was obtained at concentrations between 50 and 100 micron. These analogues also inhibited other microsomal enzymes so they apparently function as general inhibitors of microsomal enzymes.


Parental satisfaction with inpatient care of children with cerebral palsy.
Children with cerebral palsy (CP) have complex health-care needs. This study examines levels of parental satisfaction with inpatient care for children with CP at a tertiary care hospital to identify areas for improvement. Parents/guardians of children with CP and parents/guardians of children without a disability admitted to hospital completed a custom-designed questionnaire assessing six areas of the hospital admission: (i) the admission process; (ii) the child's personal care; (iii) the child's medical care; (iv) overall care of the child; (v) the parent's experience in hospital; and (vi) keeping up to date in hospital. Differences between the two groups were analysed using Student's t-tests. Parents of children with CP were significantly less satisfied with the inpatient care as compared with parents of children without a disability in four of the six categories: 'my child's personal care' (P = 0.0033), 'my child's medical care' (P = 0.0350), 'overall care' (P = 0.0081) and 'my experience in the hospital' (P = 0.0209). When the overall questionnaire was compared between the two groups, parents of children with CP were less satisfied with care than parents of children without a disability (P = 0.0036). Parents of children with CP are less satisfied with the inpatient care of their child compared with parents of children without a disability. This information should be instrumental in informing change to ensure that parent satisfaction levels improve to a level consistent with other children admitted to a tertiary care setting.


Long-term health-care utilisation in older patients with cancer and the association with the Geriatric 8 screening tool: a retrospective analysis using linked clinical and population-based data in Belgium.
Little evidence is available on the long-term health-care utilisation of older patients with cancer and whether this is associated with geriatric screening results. We aimed to evaluate long-term health-care utilisation among older patients after cancer diagnosis and the association with baseline Geriatric 8 (G8) screening results. For this retrospective analysis, we included data from three cohort studies for patients (aged ≥70 years) with a new cancer diagnosis who underwent G8 screening between Oct 19, 2009 and Feb 27, 2015, and who survived more than 3 months after G8 screening. The clinical data were linked to cancer registry and health-care reimbursement data for long-term follow-up. The occurrence of outcomes (inpatient hospital admissions, emergency department visits, use of intensive care, contacts with general practitioner [GP], contacts with a specialist, use of home care, and nursing home admissions) was assessed in the 3 years after G8 screening. We assessed the association between outcomes and baseline G8 score (normal score [>14] or abnormal [≤14]) using adjusted rate ratios (aRRs) calculated from Poisson regression and using cumulative incidence calculated as a time-to-event analysis with the Kaplan-Meier method. 7556 patients had a new cancer diagnosis, of whom 6391 patients (median age 77 years [IQR 74-82]) met inclusion criteria and were included. 4110 (64·3%) of 6391 patients had an abnormal baseline G8 score (≤14 of 17 points). In the first 3 months after G8 screening, health-care utilisation peaked and then decreased over time, with the exception of GP contacts and home care days, which remained high throughout the 3-year follow-up period. Compared with patients with a normal baseline G8 score, patients with an abnormal baseline G8 score had more hospital admissions (aRR 1·20 [95% CI 1·15-1·25]; p<0·0001), hospital days (1·66 [1·64-1·68]; p<0·0001), emergency department visits (1·42 [1·34-1·52]; p<0·0001), intensive care days (1·49 [1·39-1·60]; p<0·0001), general practitioner contacts (1·19 [1·17-1·20]; p<0·0001), home care days (1·59 [1·58-1·60]; p<0·0001), and nursing home admissions (16·7% vs 3·1%; p<0·0001) in the 3-year follow-up period. At 3 years, of the 2281 patients with a normal baseline G8 score, 1421 (62·3%) continued to live at home independently and 503 (22·0%) had died. Of the 4110 patients with an abnormal baseline G8 score, 1057 (25·7%) continued to live at home independently and 2191 (53·3%) had died. An abnormal G8 score at cancer diagnosis was associated with increased health-care utilisation in the subsequent 3 years among patients who survived longer than 3 months. Stand up to Cancer, the Flemish Cancer Society.


Adrenal cortical 11 beta-hydroxylase and side-chain cleavage enzymes. Requirement for the A- or B-pyridyl ring in metyrapone for inhibition.
The adrenal cortical enzyme systems, 11 beta-hydroxylase, P-450 11 beta, and the side-chain cleavage complex, P-450 scc, differ only in their cytochrome P-450s. Structural modifications of metyrapone, an inhibitor of cytochrome P-450 enzyme systems, have been made to determine the requirement for the A- or B-pyridyl ring for inhibition of P-45011 beta and P-450 scc activities. Three new analogs of metyrapone (A-phenylmetyrapone, B-phenylmetyrapone and diphenylmetyrapone) were synthesized and evaluated as inhibitors using a crude, defatted bovine adrenal cortical mitochondrial preparation. Characterization of the mitochondrial preparation demonstrated: enhancement of both activities by the addition of 15.0 microM adrenodoxin, the addition of 1% ethanol decreased both activities less than 10%, and the apparent Km of deoxycorticosterone for P-45011 beta was 6.8 microM and the apparent Km of cholesterol for P-450 scc was 21.6 microM. Inhibition of P-45011 beta and P-450 scc activities with these compounds demonstrated: the B-pyridyl ring of metyrapone is required for inhibition of both activities whereas requirement for the A-ring is less stringent, and the four metyrapone analogs were more selective inhibitors of P-45011 beta activity. These studies suggest that the A-phenyl metyrapone analog is a good candidate for further development of a selective adrenocortical radiopharmaceutical.


Olfactory function reflects episodic memory performance and atrophy in the medial temporal lobe in individuals at risk for Alzheimer's disease.
We examined cognitive domains and brain regions associated with olfactory performance in cognitively unimpaired older adults (CU-OAs) and individuals with or at risk for Alzheimer's Disease (AD). We compared CU-OAs (N = 55), individuals with subjective cognitive decline (SCD, N = 55), mild cognitive impairment (MCI, N = 101), and AD (N = 45) on measures of olfactory function (Brief Smell Identification Test), cognition (episodic and semantic memory), and medial temporal lobe thickness and volume. Analyses controlled for age, sex, education, and total intracranial volume. Olfactory function decreased from SCD to MCI to AD. CU-OAs outperformed all groups except SCDs on tests of cognition and olfaction. Although these measures did not differ between the CU-OAs and SCDs, olfactory function correlated with episodic memory tests and with entorhinal cortex atrophy only in the SCD group. Olfactory function also correlated with hippocampal volume and right-hemisphere entorhinal cortex thickness in the MCI group. Olfactory dysfunction reflects medial temporal lobe integrity and memory performance in a group at risk for AD with normal cognition and olfaction.


Effect of cryoprotective diluent and method of freeze-thawing on survival and acrosomal integrity of ram spermatozoa.
A multifactorial study analyzed the effects of freezing method, cryoprotective diluent, semen to diluent ratio, and thawing velocity on post-thaw motility, progressive status, and acrosomal integrity of ram spermatozoa. Although semen to diluent ratio (1:3 vs 1:6, v/v) had no effect (P greater than 0.05), overall post-thaw spermatozoal viability was highly dependent on freezing method and cryoprotectant. Improved results were obtained by freezing semen in 0.5-ml French straws compared to dry ice pelleting. Manually freezing straws 5 cm above liquid nitrogen (LN2) was comparable to cooling straws in an automated, programmable LN2 unit. Of the two cryoprotective diluents tested, BF5F (containing the surfactant component sodium and triethanolamine lauryl sulfate) yielded approximately 50% fewer (P less than 0.05) spermatozoa with loose acrosomal caps compared to TEST. Thawing straws in a water bath at a higher velocity (60 degrees C for 8 sec) had no effect (P greater than 0.05) on spermatozoal motility, progressive status ratings, or acrosomal integrity when compared to a lower rate (37 degrees C for 20 sec). For the TEST group, thawing pellets in a dry, glass culture tube promoted (P less than 0.05) percentage sperm motility at 3 and 6 hr post-thawing, but for BF5F diluted semen this approach decreased the % of spermatozoa with normal apical ridges. The results suggest that the poor fertility rates often experienced using thawed ram semen likely result not only from reduced sperm motility, but also from compromised ultrastructural integrity. This damage is expressed by an increased loosening of the acrosomal cap, a factor which appears insensitive to freezing method but markedly influenced by the cryoprotective properties of the diluents tested.


Quantitative assessment of shear-wave ultrasound elastography in thyroid nodules: diagnostic performance for predicting malignancy.
To evaluate the predictability of shear-wave ultrasound elastography (SWE) for thyroid malignancy and to compare the diagnostic performances of SWE and B-mode US. Retrospective review of 99 patients who underwent SWE before US-guided fine-needle aspiration of thyroid nodules was performed. SWE elasticity indices of the mean (Emean ), maximum (Emax), and minimum (Emin) of nodules were measured. Diagnostic performance of SWE was compared with that of B-mode US. Among a total of 99 nodules, 21 were papillary thyroid carcinoma (PTC) and 78 were benign. Emean, Emax, and Emin were significantly higher in PTCs than in benign nodules (P < 0.001). Sensitivity and specificity for predicting malignancy were 76.1 %, 64.1 % with Emax (65 kPa), 61.9 %, 76.1 % with Emin (53 kPa), and 66.6 %, 71.6 % with Emean (62 kPa). Areas under the ROC curves (Az) of SWE values were not significantly different from those of US categories on B-mode US. However, combining Emean and Emin with B-mode US of probably benign lesions improved the specificity (P = 0.02, 0.007) for predicting PTC. The quantitative parameter of SWE was significantly higher in PTC than in benign nodules, and combined use of quantitative SWE and B-mode US provided higher specificity for predicting malignancy. • Quantitative shear-wave elastography (SWE) helps differentiate benign from malignant nodules • SWE and conventional ultrasound have comparable diagnostic performance for predicting thyroid malignancy • Combined quantitative SWE and B-mode ultrasound is highly specific for thyroid malignancy.


Compassionate Use of Contezolid for the Treatment of Tuberculous Pleurisy in a Patient with a Leadless Pacemaker.
We report the case of an 87-year-old woman with tuberculous pleurisy. She developed adverse effects in the form of thrombocytopenia and gastrointestinal hemorrhage with isoniazid, and thrombocytopenia with linezolid. Her treatment was switched to contezolid plus cycloserine for a 4-week antibiotic duration, with a favorable outcome.


Malignant triton tumor of the rectum - A case report and review of the literature.
Malignant triton tumors (MTT) are rare but highly aggressive tumors that originate from the Schwann cells. These tumors can occur in any part of the body, mostly present late and carry poor prognosis. We present a 24-year-old man with a rectal MTT causing non-specific abdominal pain and recurring ileus. The MRI showed a rectal mass near the urinary bladder with compression on the seminal vesical. A complete surgical resection of the tumor was performed. The immunohistological report confirmed a rectal MTT. Because of persistent ileus during the post-operative palliative chemotherapy, another tumor debulking was performed. The patient died 9 months after the diagnosis of MTT due to local recurrence under chemotherapy. MTTs are uncommon tumors in young age with high morbidity and mortality because of local recurrence also after complete resection.


The use of agarose microwells for scalable embryoid body formation and cardiac differentiation of human and murine pluripotent stem cells.
In most pluripotent stem cell differentiation protocols the formation of embryoid bodies (EBs) is an important step. Here we describe a rapid, straightforward soft lithography approach for the preparation of hydrophilic silicon masters from different templates and the subsequent production of patterned agarose-DMEM microwell surfaces for scalable well standardized stem cell aggregation and EB formation. The non-adhesive agarose microwell plates represent an accurate replication of the templates' topography and were used for aggregation of murine induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs). Direct microscopic assessment by time-lapse analysis demonstrated rapid formation of uniformly shaped EBs from murine iPSCs with similar or even more consistent results concerning size distribution and harvesting efficiency compared to the commonly used but time-consuming hanging drop technique. For human ESCs, homogenous aggregates were obtained after single cell inoculation on agarose microwells with efficient differentiation into the cardiac lineage using state-of-the-art protocols for directed differentiation via small molecules. With this soft lithography-based strategy, sufficient and reproducible numbers of stem cell-derived cardiomyocytes necessary for tissue engineering purposes can be realized in a highly controllable manner. Moreover, it might be useful for different cell types in any application that requires scalable and highly standardized aggregation.


Prevalence of Parental Violence Toward Teachers: A Meta-Analysis.
A growing literature on parental violence toward teachers has examined the prevalence of these incidents, yet there is considerable variation across studies. There is a need for a systematic and comprehensive review to assess the extent of parent-perpetrated violence toward teachers. Using a meta-analytic approach, we examined the prevalence of violence directed against teachers by parents and how these rates vary by reporting timeframe and type of violence. We identified 5,340 articles through our initial screening process, and our final analysis included 8 studies that met criteria for this meta-analysis. Our findings show that teachers are more likely to experience non-physical forms of violence as compared to physical violence and that rates are lower as the severity and intrusiveness of the violent act increases. Implications for research and practice are discussed.


Versatility of pedicled anterolateral thigh flap in gynecologic reconstruction after vulvar cancer extirpative surgery.
Ablative surgery for vulvar cancer can involve the resection of perineum, vagina, urethra, groins, mons pubis, and abdominal wall creating complex defects. In our opinion, ALT flap is an ideal flap for reconstruction, because of low incidence of complications, long pedicle outside the radiotherapy field, capability of carrying fascia and muscle, possibility of sensate reconstruction, and low donor site morbidity. The purpose of this report is to describe our experience with ALT flap for reconstruction after vulvar cancer extirpative surgery, discussing our indications for complex defects and focusing on its versatility. We performed a retrospective review of 15 cases of complex postoncologic gynecological wounds, reconstructed with 16 pedicled ALT flap, after vulvar cancer ablative surgery between 2014 and 2016. Age of the patients ranged from 44 to 77, with mean age of 62.6. Postoperative outcomes were evaluated. The flap size ranged from 12 × 8 cm to 22 × 15 cm., in two cases we harvested vastus lateralis to fill the dead space after pelvic exenteration. In nine flaps, we could include only 1 perforator, in 7 we could use two, 5 were septocutaneous and 18 myocutaneous with a pedicle length ranging from9cm.to15cm.We had no flap necrosis.3 flaps showed dehiscence of a part of the wound that healed by dressings. In one case of obese patient the donor site showed partial wound edge necrosis, and required secondary grafting. During follow-up that ranged from 1 month to 2 years, cancer relapsed locally in 5 patients. Global performance and specific pain improved after surgery. ALT flap should be included in the first line options for reconstructions of complex defects resulting from vulvar cancer surgery because of its reliability and versatility. © 2016 Wiley Periodicals, Inc. Microsurgery 37:516-524, 2017.


Lipovitellin is a non-self recognition receptor with opsonic activity.
Lipovitellin (Lv), a glycolipoprotein, is a major component of the egg yolk, which is usually regarded as an energy reserve of nutrients essential for growth and development. We have purified Lv from ovulated eggs of the rosy barb Puntius conchonius by two-step chromatography and characterized it by staining with periodic acid/Schiff reagent and Sudan black B, amino acid composition analysis, and peptide mass fingerprinting. The results of ligand and bacterial binding assays, an enzyme-linked immunosorbent assay (ELISA), and the phagocytosis test revealed, for the first time, that the purified native form of P. conchonius Lv acts as a pattern recognition molecule with multiple specificities capable of identifying pathogen-associated molecular patterns (PAMPs), including those of lipopolysaccharide, lipoteichoic acid, and peptidoglycan, rather than self components and that it can bind Gram-negative and -positive bacteria, such as Escherichia coli and Staphylococcus aureus. These tests also showed that the P. conchonius Lv functions as an opsonin capable of enhancing macrophage phagocytosis. Taken together, these characteristics suggest that in developing embryos/larvae of P. conchonius, the native form of Lv may be physiologically involved in the sensing of invading pathogens via interaction with PAMPs and in the recruitment of the primitive macrophages that appear in early embryos to phagocytose and digest the pathogens, thereby protecting them from pathogenic attacks.


"MINES" method for genomic DNA extraction from deep biosphere biofilms.
Successful and efficient extraction of high quality, high molecular weight genomic DNA from the environmental samples is an essential primary step to understand the genetic, metabolic and evolutionary characteristics of the microbial communities. Deep mine biofilm samples that contain high amounts of mucoid exopolysaccharide often pose difficulties to obtaining refined community DNA. To circumvent this hindrance, we report our "MINES" method which we developed for optimal biofilm DNA recovery suitable for all types of high-resolution downstream applications. The method is also suitable for samples collected from landfill compost, kitchen digest (KD), and for Gram-positive Geobacillus sp. strain WSUCF1 and Gram-negative E. coli DH5α strains. In one form of the method, use of a gentle preprocessing technique to loosen the mucoid layer, combined with a multi-lytic polyzyme treatment to maximize yields from all cell types in the biofilm sample, yielded >1 μg of high molecular weight DNA (16-20 kb) per gram of the biofilm sample, with an A260/280 and A260/230 ratio of about 2. Furthermore, amplification of 16S rRNA genes as well as restriction digestion with BamHI and HindIII suggest that the newly developed method can minimize any inhibitory effects of contaminants. Results indicate that it is an appropriate methodology for the extraction of total genomic DNA for functional metagenomic studies and may be applicable to other environmental samples from which DNA extraction is challenging. IMPORTANCE: Our present knowledge of microorganisms and their enzymes from deep mine subsurfaces is based largely on laboratory studies of pure microbial cultures. These methods tend only to hit nearly 1% of the entire microbial community. In this regard, metagenomics, has emerged as a strategic approach to explore unculturable microbes through the sequencing and analysis of DNA extracted from the environmental samples. This research paper discusses our "MINES" method for genomic DNA extraction from deep biosphere biofilm samples.


Mycotic intracavernous carotid artery aneurysm in childhood.
A mycotic aneurysm of the intracavernous carotid artery which presented in an apparently classical manner during childhood, and was treated by surgery is described. The outcome was satisfactory. The literature relating to this rare condition is reviewed.


Measuring thought disorder with clinical rating scales in schizophrenic and nonschizophrenic patients.
The distinction between disorders of thought structure and disorders of thought content in a diagnostically heterogeneous psychiatric population was empirically tested using factor analysis applied to the item scores on two common clinical rating scales, the Brief Psychiatric Rating Scale and Andreasen's Thought, Language, and Communication Scale. The factor structure supported the separation of disordered thinking into disorders of thought content and disorders of thought structure or communication. Additionally, our results indicate that both aspects of disordered thinking are present in schizophrenic and manic patients; neither is specifically characteristic of schizophrenia. Indeed, disordered thought structure appears to be more common or worse in the manic patient than in the schizophrenic patient.


The association between separation individuation process and binge eating disorder in adolescents aged 12-18.
To evaluate the associations between separation individuation (SI) process and binge eating disorder (BED) in adolescence aged 12-18 years old, as most challenges related to SI process and eating disorders (EDs) tend to arise in this age group. This is a cross-sectional study of 30 adolescents diagnosed with BED and 332 healthy adolescents as a control group. All adolescents diagnosed with BED underwent clinical interviews and all adolescents in the study filled in self-reported questionnaires. Our results highlighted higher levels of need denial (p = 0.014) and rejection expectancy (p = 0.008) of SI difficulties in adolescents with BED as compared to the ones without BED. This is the first study of its nature providing good evidence for the association between SI process and BED in adolescents. Although these difficulties in SI process by themselves are not evidence of causative link, these results can contribute to the understanding of the causative factors in BED and underpin further research. The correlation can be used as a consideration in the prevention and treatment of BED. Level III, case-control analytic study.


[Current controversies in the diagnosis and management of cerebral venous and dural sinus thrombosis].
Over the last few years, the results from clinical studies and innovative radiological approaches have significantly altered the management of patients with cerebral venous thrombosis. In contrast to previously held beliefs, cerebral venous thrombosis is considered to be a relatively benign disease with an overall favourable prognosis. Mortality is <10%, and the vast majority of patients recover completely. Although the efficacy of heparin has not been unequivocally proven, it is employed in most centres as the standard therapy. There are not sufficient data supporting long-term anticoagulation. In patients with a proven hypercoagulability syndrome, anticoagulation therapy is generally recommended for a period of 6 months. Whether patients with idiopathic thrombosis should also be treated remains controversial. Non-invasive MR and CT angiography techniques have largely replaced conventional angiography for initial evaluation and follow-up examinations. Both methods have a high sensitivity for cerebral venous thrombosis. Local fibrinolytic therapy or other aggressive recanalizing methods can not be generally recommended.


Imaging analysis of the gliadin direct effect on tight junctions in an in vitro three-dimensional Lovo cell line culture system.
Tight junctions play a pivotal role in maintaining the integrity of the intestinal barrier. Their alteration is involved in the pathogenesis of celiac disease. Our aim was to investigate the gliadin effect on the tight junction proteins in an in vitro three-dimensional cell culture model through imaging analyses. Lovo multicellular spheroids were treated with enzymatically digested (PT) gliadin 500 μg/mL and its effect on actin, occludin and zonula occludens-1, was evaluated by means of confocal laser microscopy, transmission electron microscopy and image capture analysis. Compared to untreated spheroids, PT-gliadin-treated ones showed enlargement of the paracellular spaces (9.0±6.9 vs. 6.2±1.7 nm, p<0.05) at transmission electron microscopy and tight junction protein alterations at confocal microscopy and image analyses. In untreated cell cultures thickness of the fluorescence contour of actin, zonula occludens-1 and occludin appeared significantly larger and more intense than in the treated ones. In occludin planimetric analysis the lengths of the integral uninterrupted cellular contour appeared longer in untreated than in PT-gliadin treated spheroids (71.8±42.8 vs. 23.4±25.9 μm, p<0.01). Our data demonstrated that tight junction proteins are directly damaged by gliadin as shown by means of quantitative imaging analysis.


A phase II clinical trial of flutamide in the treatment of advanced breast cancer.
An antiandrogenic agent flutamide was used in the treatment of one male and 14 female postmenopausal breast cancer patients. Only a mild and transient response was seen in 2 female patients. Therefore, flutamide cannot be used solely in the treatment of female postmenopausal breast cancer.


Nipple-sparing mastectomy: A contemporary perspective.
Increasing numbers of women are undergoing nipple-sparing mastectomy, and evidence to support its use for cancer treatment and prophylaxis is expanding. An understanding of technical aspects and pitfalls of the procedure is paramount to ensure that the best results are attained. J. Surg. Oncol. 2016;113:883-890. © 2016 Wiley Periodicals, Inc.


Diagnosis of gastric cancer up to three years after negative upper gastrointestinal endoscopy.
The degree of accuracy of gastroscopy for the detection of gastric cancer is poorly understood. The aim of this retrospective study was to determine the accuracy of gastroscopy by using cancer registry records. Gastroscopic examinations (n = 37094) conducted between 1984 and 1989 were studied by linking them with hospital-based and population-based (Fukui Prefecture, Japan) cancer registry records between 1984 and 1992. False-negative gastroscopies that had been carried out within the three years preceding the diagnosis of gastric cancer were identified. The numbers of true-positive, false-positive, and false-negative examinations carried out were 659, six and 155, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were 81.0%, 100.0%, 99.1%, and 99.6%, respectively. The overall diagnostic accuracy of gastroscopy was 99.6%. There was little difference in sensitivity results between the patient groups with regard to reason for referral, type of endoscope used, experience of endoscopist, or location of gastric cancer. The percentage of tumours representing early gastric cancer, identified after false-negative gastroscopy, was lower for those situated in the cardia or gastric body than for those in the angular notch or the antrum. The accuracy of gastroscopy in the detection of gastric cancer is satisfactory, but false-negative results are sometimes obtained. We emphasize the importance of repeated endoscopic examination for the detection of gastric cancer.


Detection of the basic fibroblast growth factor low affinity binding site in cardiac sarcolemmal vesicles.
Bovine cardiac sarcolemmal (SL) vesicles contain basic fibroblast growth factor (bFGF)-binding proteins. Binding to native SL vesicles was specific and saturable with a Kd of 6.9 nM and a Bmax of 15.2 pmoles bFGF/mg vesicle protein. Using radioiodinated bFGF as a probe, autoradiography of SL proteins subjected to SDS-PAGE and electroblotting onto nitrocellulose revealed a set of 3-4 bands, of an apparent molecular weight of 100-150 kDa. bFGF binding to these bands was reduced by pretreatment of SL vesicles with heparinase. Binding was abolished by treatment of blot strips with heparinase or high salt concentrations (greater than 0.6 NaCl) but not endoglycosidase F. bFGF-binding activity remained associated with the membrane fraction following an alkaline wash, which removed peripheral membrane proteins. These data suggest that the cardiac SL contains an integral proteoglycan(s) which may be a low affinity binding/storage site of endogenous bFGF.


Parenting, the other oldest profession in the world - a cross-sectional study of parenting and child outcomes in South Africa and Malawi.
Parenting quality is important in child development. In the presence of HIV poverty and life stress, parenting may be challenged and child development affected. This study examines cross-sectional associations of situational factors such as poverty, mental health, HIV status, living with a biological parent, and stigma with good parenting and child outcomes (n=989; age=4-13 years) within the Child Community Care study (South Africa and Malawi). A parenting measure was created from 10 variables comprising 6 child and 4 parent ratings. These were highly correlated. Total parenting score was generated on a 10 point continuous scale, with a good parenting cut off then defined as >=8 out of a possible 10. Five factors were associated with good parenting. Positively associated with good parenting were being the biological parent of the child, parental mental health and dwelling in households with multiple adults. Poverty and stigma were negatively associated with good parenting. Using multiple mediation analysis, a positive direct effect of good parenting was found on child self-esteem, child behaviour and educational risks with a partial mediation via child depression and trauma. These data highlight possible intervention points. Influences on parenting could be seen through being the biological parent, parental mental health, poverty and stigma. In these challenging environments, health, nutrition, mental health, education, and treatment to keep parents alive are all clearly identified as potential pathways to ensure child well-being.


Cytokines and bone loss in a 5-year longitudinal study--hormone replacement therapy suppresses serum soluble interleukin-6 receptor and increases interleukin-1-receptor antagonist: the Danish Osteoporosis Prevention Study.
The proinflammatory cytokines interleukin-1 beta (IL-1 beta) and IL-6 may play a central role in the acceleration of postmenopausal bone loss, but observational studies have led to contradictory results. Estrogen-dependent changes in the production of IL-1 receptor antagonist (IL-1ra) and the soluble IL-6 receptor (sIL-6R) potentially modify cytokine bioactivity. We therefore assessed the impact of menopause and hormone replacement therapy (HRT) on cytokines and activity modifiers in serum within a 5-year longitudinal study. One hundred sixty perimenopausal women (age 50.1 +/- 2.8 years) were randomized to HRT or no treatment. Serum IL-6 increased with age (r = 0.16; p < 0.05), but cytokines did not correlate with baseline bone mineral density (BMD). HRT led to small increases in IL-1ra (p < 0.001) and IL-6 (p < 0.05), with a decrease in sIL-6R (p < 0.01) and no change in IL-1 beta. No changes were observed in the control group. IL-1ra was inversely correlated with bone loss at the ultradistal forearm (r = 0.29; p < 0.05) and to a lesser degree at the spine (r = 0.20; p = 0.09). In addition, there was a weak positive correlation between sIL-6R and bone loss at the ultradistal forearm (r = 0.26; p < 0.05). High IL-6 levels were associated with slower bone loss (spine r = 0.31, p < 0.01) and controlling for age did not diminish this association. The percent change in sIL-6R during HRT was correlated with the bone loss at the femoral neck (r = -0.29; p < 0.01) and weakly with bone loss in the spine (r = -0.16; p = 0.17). In conclusion, serum IL-1ra and sIL-6R are influenced by HRT and are associated with the rate of bone loss in perimenopausal women.


The non-competitive AMPA receptor antagonist (GYKI 52466) blocks quisqualate-induced acetylcholine release from the rat hippocampus and striatum: an in vivo microdialysis study.
The effects of the non-N-methyl-D-aspartate (NMDA) agonist quisqualate (QUIS) and selective AMPA/kainate receptor antagonist 1-(aminophenyl)-methyl-7, 8-methyilendioxy-5H-2,3-benzodiazepine (GYKI 52466) on the release of acetylcholine (ACh) from the hippocampus and striatum of freely moving rats were studied by transversal microdialysis. Acetylcholine level in the dialisate was measured by the high performance liquid chromatography (HPLC) method with an electrochemical detector. The QUIS (100 microM) perfused through the striatum induced an increase of extracellular ACh level (250%) which lasted for over 1h and gradually returned to basal values. Local perfusion of GYKI 52466 (10-100 microM) to the striatum did not change the basal release of ACh. GYKI 52466 (10 microM) administered together with QUIS (100 microM) in he striatum antagonized the stimulant effect of QUIS on the ACh release. Local administration of the QUIS (100 microM) through the microdialysis fiber implanted in the hippocampus, caused a long lasting increase of extracellular hippocampal ACh level (360%) which was reversed when the drug was withdrawn from the perfusion solution. The stimulant effect of QUIS was antagonized by concomitant perfusion of GYKI (10 microM). No effect was seen on the basal ACh release when GYKI (10-100 microM) was perfused through the hippocampus. Local perfusion with tetrodotoxin (1 microM) decrease the basal release of ACh and prevented the QUIS-induced increase of ACh both in the hippocampus and striatum. Our in vivo neurochemical results indicate that hippocampal and striatal cholinergic systems are regulated by non-NMDA (probably AMPA) glutamatergic receptors located in the hippocampus and striatum.


Succinate-Directed Approaches for Warburg Effect-Targeted Cancer Management, an Alternative to Current Treatments?
Approximately a century ago, Otto Warburg discovered that cancer cells use a fermentative rather than oxidative metabolism even though the former is more inefficient in terms of energy production per molecule of glucose. Cancer cells increase the use of this fermentative metabolism even in the presence of oxygen, and this process is called aerobic glycolysis or the Warburg effect. This alternative metabolism is mainly characterized by higher glycolytic rates, which allow cancer cells to obtain higher amounts of total ATP, and the production of lactate, but there are also an activation of protumoral signaling pathways and the generation of molecules that favor cancer progression. One of these molecules is succinate, a Krebs cycle intermediate whose concentration is increased in cancer and which is considered an oncometabolite. Several protumoral actions have been associated to succinate and its role in several cancer types has been already described. Despite playing a major role in metabolism and cancer, so far, the potential of succinate as a target in cancer prevention and treatment has remained mostly unexplored, as most previous Warburg-directed anticancer strategies have focused on other intermediates. In this review, we aim to summarize succinate's protumoral functions and discuss the use of succinate expression regulators as a potential cancer therapy strategy.


Road traffic accidents and diabetes: insulin use does not determine risk.
Progressive restrictions placed on insulin-treated patients with diabetes exclude them from driving group 2 and class C1 and D1 vehicles. This reflects an assumption that an increased risk of hypoglycaemia in these patients will cause road traffic accidents. These restrictions have been implemented without any consistent evidence that this is the case. The aim of the study was therefore to investigate whether the rate of road traffic collisions in insulin-treated patients was higher than that of the non-diabetic population using a population register-based study. A historical cohort study combined information from the Devon and Cornwall Constabulary database on road traffic collisions with the district wide retinal screening database, to provide an anonymized matched database of road traffic collisions in the diabetic population. Accident rates were calculated in the diabetic population and compared to rates in the non-diabetic population using relative risks. The estimated overall annual accident rate for the non-diabetic population was 1469 per 100,000 vs. 856 per 100,000 for the diabetic population as a whole (Chi-squared, P < 0.001). On stratification of the groups by age, within the insulin-treated group there was no significant difference in the accident rate compared to the non-diabetic population, with relative risks between 0.51 [confidence interval (CI) 0.25-1.05] and 1.13 (CI 0.88-1.46). Our findings suggest that insulin-treated patients as a group do not pose an increased risk to road safety. They reiterate the need for an individualized risk-based assessment when considering driving restrictions.


γ Actinorhodin a natural and attorney source for synthetic dye to detect acid production of fungi.
Colors from natural sources are gaining popularity because synthetic colors are carcinogenic. Natural colors are obtained from plants or microorganisms. Pigments produced by microorganisms have advantages over plant pigments, due to their ease of use and reliability. In the present study, a blue pigment producing actinomycete klmp33 was isolated from the Gulbarga region in India. The isolate was identified based on morphologic, microscopic, and biochemical characterization, and 16S rRNA sequencing. Phylogenetic analysis of the isolates showed a close relationship with Streptomyces coelicolor. Pigment produced by the isolate was analyzed using UV-visible spectroscopy, thin-layer chromatography, Fourier transform infrared and liquid chromatography/mass spectroscopy analysis, and was identified as γ actinorhodin. γ-Actinorhodin is used as a pH indicator which deviates from acid to non-acid. Moreover, it subrogates synthetic dye.


The myeloma landscape in 2008.
Highlights From the American Society of Hematology 2007 Annual Meeting, Atlanta, Georgia, December 8-11, 2007.


Suppressive effect of interleukin-1 on pulmonary cytochrome P450 and superoxide anion production.
Interleukin-1 has been shown to prolong the survival of rats exposed to lethal concentrations of oxygen. This oxygen tolerance has been attributed by some workers to an increase of manganese superoxide dismutase. We report here that the administration of interleukin-1 to male adult rats results in (i) significant decrease of pulmonary cytochrome P450 at 24 and 72 hours, (ii) decrease of P450 IIB1 mRNA at 24 and 72 hours and (iii) significant decrease of superoxide anion generation from pulmonary microsomes isolated from treated rats. To the best of our knowledge, this is the first report to demonstrate these effects of interleukin-1 on pulmonary P450 and its oxidase activity (O2- generation). On the basis of these results and several earlier reports in which various P450 depressants have been shown to depress superoxide production from microsomes and to prolong the lives of rodents in hyperoxia, we conclude that oxygen tolerance induced by interleukin-1 administration is likewise mediated, at least in part, by reduced generation of superoxide anion from cytochrome P450 monooxygenase system.


Scalable Synthesis of One-Dimensional Mesoporous ZnMnO3 Nanorods with Ultra-Stable and High Rate Capability for Efficient Lithium Storage.
The cost-efficient ZnMnO3 has attracted increasing attention as a prospective anode candidate for advanced lithium-ion batteries (LIBs) owing to its resourceful abundance, large lithium storage capacity and low operating voltage. However, its practical application is still seriously limited by the modest cycling and rate performances. Herein, a facile design to scalable synthesize unique one-dimensional (1D) mesoporous ZnMnO3 nanorods (ZMO-NRs) composed of nanoscale particles (≈11 nm) is reported. The 1D mesoporous structure and nanoscale building blocks of the ZMO-NRs effectively promote the transport of ions/electrons, accommodate severe volume changes, and expose more active sites for lithium storage. Benefiting from these appealing structural merits, the obtained ZMO-NRs anode exhibits excellent rate behavior (≈454 mAh g-1 at 2 A g-1 ) and ultra-long term cyclic performance (≈949.7 mAh g-1 even over 500 cycles at 0.5 A g-1 ) for efficient lithium storage. Additionally, the LiNi0.8 Co0.1 Mn0.1 O2 //ZMO-NRs full cell presents a practical energy density (≈192.2 Wh kg-1 ) and impressive cyclability with approximately 91 % capacity retention over 110 cycles. This highlights that the ZMO-NRs product is a highly promising high-rate and stable electrode candidate towards advanced LIBs in electronic devices and sustainable energy storage applications.


Does pre-exposure to warming conditions increase Mytilus galloprovincialis tolerance to Hg contamination?
The degree to which marine invertebrate populations can tolerate extreme weather events, such as short-term exposure to high temperatures, and the underlying biochemical response mechanisms are not yet fully understood. Furthermore, scarce information is available on how marine organisms respond to the presence of pollutants after exposure to heat stress conditions. Therefore, the present study aimed to understand how the mussel Mytilus galloprovincialis responds to Hg pollution after pre-exposure to warming conditions. Mussels were exposed to control (17°C) and warming (21°C) conditions during 14days, followed by Hg contamination during 28days under different temperature regimes (17 and 21°C). The results obtained demonstrated significantly higher Hg concentrations in mussels under 17°C during the entire experiment than in organisms exposed to 21°C during the same period, which resulted in higher oxidative stress in mussels under control temperature. Significantly higher Hg concentrations were also observed in mussels pre-exposed to 21°C followed by a 17°C exposure comparing with organisms maintained the entire experiment at 21°C. These results may be explained by higher metabolic capacity in organisms exposed to 17°C after pre-exposure to 21°C that although induced antioxidant defences were not enough to prevent oxidative stress. No significant differences in terms of Hg concentration were found between mussels exposed to 17°C during the entire experiment and organisms pre-exposed to 21°C followed by a 17°C exposure, leading to similar oxidative stress levels in mussels exposed to both conditions. Therefore, our findings demonstrated that pre-exposure to warming conditions did not change mussels' accumulation and tolerance to Hg in comparison to Hg contaminated mussels maintained at control temperature. Furthermore, the present study indicate that organisms maintained under warming conditions for long periods may prevent the accumulation of pollutants by decreasing their metabolism which will limit cellular injuries. Mussels under warming conditions presented reduced metabolic capacity, resulting in lower Hg accumulation, which in turn prevented higher damages and, consequently, physiological impairments.


Involvement of bradykinin, cytokines, sympathetic amines and prostaglandins in formalin-induced orofacial nociception in rats.
1. This study characterises some of the mechanisms and mediators involved in the orofacial nociception triggered by injection of formalin into the upper lip of the rat, by assessing the influence of various treatments on behavioural nociceptive responses (duration of facial rubbing) elicited either by a low subthreshold (i.e. non-nociceptive; 0.63%) or a higher concentration of the algogen (2.5%). 2. The kininase II inhibitor captopril (5 mg kg(-1), s.c.) and prostaglandin(PG) E(2) (100 ng lip(-1)) potentiated both phases of the response to 0.63% formalin, whereas tumour necrosis factor (TNF alpha; 5 pg lip(-1)), interleukin(IL)-1 beta (0.5 pg lip(-1)), IL-6 (2 ng lip(-1)) and IL-8 (200 pg lip(-1)), or the indirectly acting sympathomimetic drug tyramine (200 microg lip(-1)), each augmented only the second phase of nociception. 3. Conversely, both phases of nociception induced by 2.5% formalin were inhibited by the bradykinin (BK) B(2) receptor antagonist HOE140 (5 microg lip(-1)) or the selective beta(1)-adrenoceptor antagonist atenolol (100 microg lip(-1)). However, the BK B(1) receptor antagonist des-Arg(9)-Leu(8)-BK (1 and 2 microg lip(-1)), antibody and/or antiserum against each of the cytokines, the adrenergic neurone blocker guanethidine (30 mg kg(-1) day(-1), s.c., for 3 days) and the cyclooxygenase(COX)-2 inhibitor celecoxib (50 and 200 microg lip(-1), s.c.; or 1 and 3 mg kg(-1), i.p.) reduced only the second phase of the response. The nonselective COX inhibitor indomethacin and the 5-lipoxygenase activating protein inhibitor MK886 did not change formalin-induced nociception. 4. Our results indicate that BK, TNF-alpha, IL-1 beta, IL-6, IL-8, sympathetic amines and PGs (but not leukotrienes) contribute significantly to formalin-induced orofacial nociception in the rat and the response seems to be more susceptible to inhibition by B(2) receptor antagonist and selective COX-2 inhibitor than by B(1) receptor antagonist or nonselective COX inhibitor.


Can early measles vaccination control both measles and respiratory syncytial virus infections?
Measles virus and respiratory syncytial virus (RSV) are two important global health pathogens causing substantial morbidity and mortality worldwide. The current measles vaccination schedule has the first dose given at 9-12 months of age and the second dose given at 15-18 months of age. Measles outbreaks have been associated with an increase in severe RSV infections in children younger than 6 months, probably as a result of measles-induced immunosuppression. A resurgence in measles cases was already occurring before the COVID-19 pandemic, which has affected global immunisation programmes, resulting in millions of children, mostly in low-income and middle-income countries (LMICs), missing out on their measles vaccine. This will leave many children living in the most vulnerable of circumstances highly susceptible to measles and RSV infections when current COVID-19 public health control measures are lifted. This Viewpoint discusses these issues and highlights the need for urgent action to address this looming crisis. The use of early measles vaccination at 4 months of age could be an effective strategy to prevent severe morbidity and death from both measles and RSV infections in many LMICs.


Simulated effects of tidal inundation and light reduction on Zostera muelleri flowering in seagrass nurseries.
Zostera muelleri is an abundant seagrass species distributed through intertidal and shallow subtidal waters on the subtropical coasts of Australia. The vertical distribution of Zostera is likely defined by tidal influences, particularly desiccation and light reduction stresses. These stresses were expected to affect the flowering of Z. muelleri; however, it is difficult to quantify the effects of tidal inundation with field studies due to multiple confounding environmental factors affecting flowering (e.g., water temperature, herbivory, nutrients). A laboratory aquarium experiment compared the effects of two levels of tidal height (intertidal and subtidal) and light intensity (shaded and unshaded) on flowering timing, abundance, the ratio between flowering shoots and vegetative shoots, the morphology and duration of flower development. The earliest and greatest flowering intensity was recorded in the subtidal-unshaded group, with no flowers observed in the intertidal-shaded group. Notably, the peak flowering time was the same across shaded and unshaded treatments. Shading prolonged the timing of the first flowering and reduced the density of flowering shoots and spathes, while tidal inundation had a more significant effect on the density of flowering shoots and the density of spathes. Results showed that Z. muelleri could flower under low light conditions or tidal stress but not when exposed to both stresses simultaneously in a laboratory 'nursery setting'. Therefore, applying subtidal-unshaded conditions appears to be beneficial for seagrass nurseries aimed at improved flower abundance despite the plants previously being collected from and adapted to intertidal meadows. Further studies that explore the suitable conditions for triggering and optimising the flowering will be beneficial in designing cost-effective seagrass nurseries.


The Impact of Using Different Methods to Assess Completeness of 24-Hour Urine Collection on Estimating Dietary Sodium.
The standard for population-based surveillance of dietary sodium intake is 24-hour urine testing; however, this may be affected by incomplete urine collection. The impact of different indirect methods of assessing completeness of collection on estimated sodium ingestion has not been established. The authors enlisted 507 participants from an existing community study in 2009 to collect 24-hour urine samples. Several methods of assessing completeness of urine collection were tested. Mean sodium intake varied between 3648 mg/24 h and 7210 mg/24 h depending on the method used. Excluding urine samples collected for longer or shorter than 24 hours increased the estimated urine sodium excretion, even when corrections for the variation in timed collections were applied. Until an accurate method of indirectly assessing completeness of urine collection is identified, the gold standard of administering para-aminobenzoic acid is recommended. Efforts to ensure participants collect complete urine samples are also warranted.


A Ca2+-permeable non-selective cation channel activated by depletion of internal Ca2+ stores in single rabbit portal vein myocytes.
In vascular smooth muscle cells many agonists cause the release of Ca2+ ions from internal stores. An important problem concerns the mechanism by which the intracellular stores are refilled subsequent to depletion. In the present study, we describe the properties of a Ca2+-permeable non-selective cation channel current that is activated in rabbit portal vein myocytes by depletion of internal Ca2+ stores. Application of cyclopiazonic acid (CPA), which depletes internal Ca2+ stores, activated whole-cell currents that had a reversal potential (E(r)) of about +50 mV in 1.5 mM external Ca2+ (Ca2+o). In 0 mM Ca2+o, the currents were larger and E(r) was approximately 0 mV. Application of CPA and caffeine during cell-attached recording activated single inward channel currents at negative potentials, which had a slope conductance of 2-3 pS and an E(r) of +20 mV. The slope conductance in 0 and 110 mM Ca2+o was 7 and 1.5 pS, respectively, and E(r) values indicated that these non-selective cation channels are highly permeable to Ca2+ ions. Bath application of the cell-permeant Ca2+ chelator, BAPTA-AM, also activated similar currents, indicating that these channels are not activated by Ca2+. Spontaneous channel currents with similar properties to store-operated channels were observed in some patches. Application of W-7, an inhibitor of the Ca2+-binding protein calmodulin, also activated similar Ca2+-permeable channel currents. In conclusion, it is demonstrated that agents that deplete Ca2+ stores and inhibit calmodulin binding activate Ca2+-permeable non-selective cation channel currents in rabbit portal vein myocytes. These channels may have an important role in vascular smooth muscle in providing an influx of Ca2+ to refill depleted internal Ca2+ stores and appear to possess different characteristics to store-operated channels described in other vascular smooth muscle preparations.


A review of studies on blood lead concentrations of traditional Mexican potters.
Traditional Mexican potters and their families have been occupationally exposed to lead for centuries; however, studies on blood lead levels (BLL) and their adverse health impact on this population are scarce. There is no safe BLL, even at 1 μg/dL there are associated health effects. To systematize and characterize Mexican potters' historic lead exposure through their BLL and associated health outcomes. Using PRISMA guidelines, we conducted a systematic review through January 2021 of published studies on BLL and associated health outcomes in Mexican potters. Fifteen studies containing data from 1980 to 2013 met the inclusion criteria and were published between 1980 and 2018. Study populations ranged from n = 5 to n = 457, and included adult potters (mean BLL 37.9 ± 16.2 μg/dL) and/or their children (mean BLL 22.5 ± 10.5 μg/dL). Studies reported on general lead poisoning symptoms, neurotoxic and nephrotoxic outcomes as well as correlated biomarkers. Our results confirm high occupational and para-occupational lead exposure. Despite governmental and non-governmental initiatives to promote lead-free glazes, lead continues to be used by traditional potters and their families.


Acetylcholinesterase mutation in diazinon-resistant Haematobia irritans (L.) (Diptera: Muscidae).
Acetylcholinesterase (AChE) cDNA from individual field-collected diazinon-resistant horn flies was amplified by RT-PCR. Sequencing of the amplification products revealed that 8/12 of the diazinon-resistant horn flies contained a point mutation previously associated with resistance to organophosphates in house flies and Drosophila, strongly suggesting that this cDNA encodes the AChE that is the target site for organophosphate (OP) pesticide. The point mutation (G262A) resulted in a shift from glycine to alanine in the mature HiAChE amino acid sequence at position 262. Allele-specific PCR and RLFP assays were developed to diagnose the presence or absence of the G262A mutation in individual flies. Use of the allele-specific assays each demonstrated the presence of the G262A mutation in 10 of 12 individual field-collected flies, demonstrating higher sensitivity than direct sequencing of RT-PCR amplification products. The G262A mutation was found in additional fly populations previously characterized as OP-resistant, further supporting that this AChE is the target site for OP pesticide. The allele-specific assay is a useful tool for quantitative assay of the resistance allele in horn fly populations.


Size-exclusion chromatography using deuterated mobile phases.
The effect of deuterated solvents in size-exclusion chromatography (SEC) was studied by comparing intrinsic viscosity measurements, SEC calibration curves, and column efficiency using water-soluble polymers. For aqueous SEC, the use of deuterium oxide slightly increases the SEC elution volume. To verify that adsorption onto the packing was absent, data from exclusion experiments were compared at 35 and 50 degrees C. Our results indicate that adsorption is not occurring for pullulan or polyethylene glycol (PEG)/poly(ethylene oxide) (PEO); for the latter, however, the elution volume increased using both D2O and H2O, indicative of slight hydrodynamic volume contraction of PEG/PEO at higher temperatures. A moderate increase in band broadening (moderate decrease in column efficiency) was observed using D2O. Finally, the effects of chloroform versus deuterated chloroform were evaluated, but no hydrodynamic volume changes were observed.


Comparative ecotoxicity of three polluted industrial soils for the Collembola Folsomia candida.
We tried to quantify the ecotoxicity of three industrial soil samples contaminated by different metals, using the Collembolan Folsomia candida as a biological model and mortality, growth and reproduction as parameters. The observed ecotoxicities are rather normal for the first such soil sample (aluminum factory) but are high for the second sample (ore treatment) and relatively low for the third one (zinc factory) considering its high metal concentrations. For these last two soil samples, an unusual ecotoxicity plotting is observed: two high ecotoxicity recordings fit with a low and high percentage of polluted soil to non-polluted soil and noticeably lower ecotoxicity recordings are observed between them. Chemical analyses of metals in pore waters show that arsenic probably explains part of such an unusual ecotoxicity curve. Otherwise, mortality and growth of the animals are less sensitive parameters than reproduction. Our experiments show that the results of the ecotoxicological assays of polluted soils are complex and difficult to interpret.