Nanoparticle Uptake in Plants: Gold Nanomaterial Localized in Roots of Arabidopsis thaliana by X-ray Computed Nanotomography and Hyperspectral Imaging.
Terrestrial plants can internalize and translocate nanoparticles (NPs). However, direct evidence for the processes driving the NP uptake and distribution in plants is scarce at the cellular level. Here, NP-root interactions were investigated after 10 days of exposure of Arabidopsis thaliana to 10 mg·L-1 of negatively or positively charged gold NPs (∼12 nm) in gels. Two complementary imaging tools were used: X-ray computed nanotomography (nano-CT) and enhanced dark-field microscopy combined with hyperspectral imaging (DF-HSI). The use of these emerging techniques improved our ability to detect and visualize NP in plant tissue: by spectral confirmation via DF-HSI, and in three dimensions via nano-CT. The resulting imaging provides direct evidence that detaching border-like cells (i.e., sheets of border cells detaching from the root) and associated mucilage can accumulate and trap NPs irrespective of particle charge. On the contrary, border cells on the root cap behaved in a charge-specific fashion: positively charged NPs induced a higher mucilage production and adsorbed to it, which prevented translocation into the root tissue. Negatively charged NPs did not adsorb to the mucilage and were able to translocate into the apoplast. These observations provide direct mechanistic insight into NP-plant interactions, and reveal the important function of border cells and mucilage in interactions of plants with charged NPs.


Long-Term Efficacy, Safety, and Durability of Cabotegravir and Rilpivirine as 2-Drug Oral Maintenance Therapy After 6 Years of Study.
In the LATTE study, daily oral cabotegravir + rilpivirine demonstrated higher rates of efficacy than efavirenz + 2 nucleoside reverse-transcriptase inhibitors (NRTIs) through Week 96 in antiretroviral therapy (ART)-naive adults with human immunodeficiency virus (HIV)-1. We present the results from 6 years of continued treatment with oral cabotegravir + rilpivirine. LATTE was a phase IIb, randomized, multicenter, partially blinded, dose-ranging study in ART-naive adults with HIV-1. After a 24-week induction phase with cabotegravir + 2 NRTIs, participants with HIV-1 ribonucleic acid (RNA) <50 copies/mL were randomized to receive cabotegravir (10, 30, or 60 mg) + rilpivirine (25 mg) in the maintenance phase through Week 96 and switched to cabotegravir 30 mg + rilpivirine 25 mg in the open-label phase through Week 312. Of 160 participants who entered the maintenance phase, 111 completed the study at Week 312. At Week 312, 105 (66%) participants maintained HIV-1 RNA <50 copies/mL, 15 (9%) had HIV-1 RNA ≥50 copies/mL, and 40 (25%) had no virologic data. Eight participants met protocol-defined virologic failure criteria through Week 312, 2 of whom met protocol-defined virologic failure criteria after Week 144. Six participants developed treatment-emergent resistance to 1 or both agents during the study, 3 of whom developed integrase inhibitor resistance substitutions. Two participants (1%) reported drug-related serious adverse events. Few adverse events led to withdrawal during the open-label phase (n = 5, 3%). Oral cabotegravir + rilpivirine demonstrated efficacy in the majority of participants and an acceptable safety profile through 6 years of treatment, demonstrating its durability as maintenance therapy for HIV-1.


Immobilization of peptides with distinct biological activities onto stem cell culture substrates using orthogonal chemistries.
We have used the orthogonal carbodiimide condensation and copper-catalyzed azide-alkyne "click" cycloaddition (CuAAC) reactions to prepare self-assembled monolayers that present distinct peptides to stem cells in a bioinert background. The approach involved first forming mixed SAMs with three components: (i) an azide-terminated hexaethylene glycol alkanethiolate (HS-EG6-N3), (ii) a carboxylate-terminated hexaethylene glycol alkanethiolate (HS-EG6-COOH), and (iii) a triethylene glycol alkanethiolate (HS-EG3). An acetylene-bearing peptide and an amine-terminated peptide were then immobilized to these substrates using a "click" CuAAC reaction and a carbodiimide condensation reaction, respectively. Polarization-modulated infrared reflectance-absorbance spectroscopic analysis demonstrated formation of well-ordered, close-packed self-assembled monolayers (SAMs), chemoselective conjugation of amine-terminated peptides to surface carboxylate groups, and subsequent conjugation of acetylene-terminated peptides to the azide groups on SAMs. Varying the mole fraction of HS-EG6-N3, HS-EG6-COOH, and HS-EG3 during SAM formation allowed for control over the densities of each peptide on the substrate. Substrates presenting varying surface densities of RGESP (a nonfunctional peptide), RGDSP (a cell adhesion peptide), or TYRSRKY (a heparin/heparan sulfate-binding peptide) were then used to characterize the relationship between peptide surface density and human mesenchymal stem cell (hMSC) adhesion. Results demonstrate that RGESP does not influence RGDSP-mediated adhesion of hMSCs, which indicates that a second peptide with distinct bioactivity can be immobilized alongside RGDSP to characterize the influence of two peptides on hMSC behavior. Our results also demonstrate that RGDSP and TYRSRKY act synergistically to promote hMSC adhesion in the absence of serum. Interestingly, heparin sequestered by TYRSRKY inhibits cell adhesion on substrates presenting RGDSP = 0.1% and > or = 0.1% TYRSRKY or RGDSP = 1% and > or = 0.5% TYRSRKY. Taken together, these results indicate that two peptides can be controllably presented to stem cells on the same otherwise bioinert SAM substrate, and that multiple, distinct extracellular moieties act in concert to regulate hMSC adhesion.


A Dietary Pattern of Higher Fish, Egg, Milk, Nut, Vegetable and Fruit, and Lower Salt Intake Correlates With the Prevalence and Control of Hypertension.
The effects of food on the prevalence and control of hypertension are unclear. We aimed to investigate whether a dietary pattern of higher fish, egg, milk, nut, vegetable and fruit consumption, and lower salt intake was associated with hypertension in China. A total of 15,303 subjects were recruited from September 2012 to December 2014. Groups with (n = 1,604) and without (n = 13,660) hypertension were formed for a case-control study. The hypertensive participants were classified into the controlled blood pressure (BP) subgroup (n = 397) and the uncontrolled BP subgroup (n = 1,207). Data on the average weekly intake of fish, eggs, milk, nuts, vegetables, fruit, and salt in the past year were collected. Higher intake was defined as greater than or equal to median food intake. Higher fish, egg, milk, nut, vegetable, and fruit intake correlated with lower hypertension prevalence, and fish and fruit intake were the strongest associated factors. Meanwhile, higher fruit intake, the highest quartile of egg or milk intake, and the lowest quartile of salt intake correlated with better BP control. Furthermore, the dietary pattern was associated with lower hypertension prevalence (odds ratio [OR]: 0.88, 95% confidence interval [CI]: 0.84-0.92; P < 0.001) and better BP control (OR: 1.11, 95% CI: 1.03-1.21; P = 0.011). However, the dietary pattern did not correlate with BP control after excluding fruit intake. The dietary pattern correlated with lower hypertension prevalence and better BP control, and its association with BP control might be driven by higher fruit consumption.


Taxanes in combination with platinum derivatives for the treatment of ovarian cancer during pregnancy: A literature review .
Ovarian cancer is one of the most common types of solid carcinoma diagnosed during pregnancy. Taxane plus a platinum derivative is a combination therapy that is predominantly used in the treatment of ovarian cancer in non-pregnant women. Pregnancy adds various complexities to a course of treatment. In pregnant patients diagnosed with cancer during the first trimester, the risks of fetal malformations and fetal loss increase following the administration of cytotoxic drugs, and this is higher with multi-agent vs. single-agent chemotherapy (~ 25 vs. 10%). Exposure during the second and third trimester has little influence on teratogenic effects but increases the risk of intrauterine growth retardation, prematurity, low birth weight, and bone marrow toxicity. The present study aimed to review the maternal and fetal safety of treatment with taxane plus platinum derivatives for ovarian cancer during pregnancy. Relevant literature was retrieved from the Embase and PubMed databases using the search terms "ovarian cancer", "pregnancy", "taxane", "paclitaxel", "docetaxel", "platinum", "cisplatin", and "carboplatin". All available data up until September 2016 was synthesized, with no language restrictions. A total of 11 articles (including 13 pregnancies and 14 newborns) were retrieved that reported on the use of standard-dose taxane and platinum chemotherapy, including 9 cases treated with paclitaxel and carboplatin, 3 cases treated with paclitaxel and cisplatin, and 1 case treated with docetaxel and cisplatin. In 13 of the 14 (92.9%) births included, a healthy neonate was born, with follow-up ranging from 2 to 160 months. The average weight of the neonates at the time of delivery was 2,442.1 g. In 7 of 9 the case reports that provided survival data, the mother was alive and disease-free at the end of follow-up (ranging from 2 to 40 months). In conclusion, combination therapy with taxanes and a platinum derivative may play a significant role in the management of pregnant patients with ovarian cancer during the second and third trimester. Exposure to this combination of agents during the second and third trimester does not appear to have a significant bearing on fetal mortality and abortion. .


Supramolecular protection from the enzymatic tyrosine phosphorylation in a polypeptide.
Here we report two new artificial pseudopeptidic cages that bind the EYE peptide epitope in pure water at physiological pH (as studied by fluorescence and NMR spectroscopies). The supramolecular complexation of the Tyr residues efficiently precludes their subsequent PTK-catalysed phosphorylation. Our results show a supramolecular modulation of the PTK activity by competitive substrate caging.


Utilising established SDL-screening methods as a tool for the functional genomic characterisation of model and non-model organisms.
The trend for large-scale genetic and phenotypic screens has revealed a wealth of information on biological systems. A major challenge is understanding how genes function and putative roles in networks. The majority of current gene knowledge is garnered from studies utilising the model yeast Saccharomyces cerevisiae. We demonstrate that synthetic dosage lethal genetic array methodologies can be used to study genetic networks in other yeasts, namely the fungal pathogen Candida glabrata, which has limited forward genetic tools, due to the lack of 'natural' mating. We performed two SDL screens in S. cerevisiae, overexpressing the transcriptional regulator UME6 as bait in the first screen and its C. glabrata ortholog CAGL0F05357g in the second. Analysis revealed that SDL maps share 204 common interactors, with 10 genetic interactions unique to C. glabrata indicating a level of genetic rewiring, indicative of linking genotype to phenotype in fungal pathogens. This was further validated by incorporating our results into the global genetic landscape map of the cell from Costanzo et al. to identify common and novel gene attributes. This data demonstrated the utility large data sets and more robust analysis made possible by interrogating exogenous genes in the context of the eukaryotic global genetic landscape.


Differences in the metabolic and hormonal response to exercise between racing cyclists and untrained individuals.
1. Six well-trained cyclists and six untrained subjects were studied during and immediately after four successive 7 min periods of exercise at 30, 45, 60 and 75% of their maximal work capacity. 2. Venous blood samples were taken at rest, at the end of each exercise period and 5 min following the end of exercise, for estimation of metabolites in blood and plasma insulin, growth hormone, cortisol and catecholamines. 3. The results showed significant differences in the mobilization and utilization of muscle fuels between the athletically fit cyclists and the untrained group. In the cyclists, glucose, glycerol and free fatty acid concentrations were higher, but lactate, pyruvate and alanine were lower than in the untrained subjects during exercise. 4. Plasma catecholamines rose in both groups during exercise but the rise was significantly less in the racing cyclists. Plasma insulin was depressed to a greater extent in the untrained subjects during exercise and plasma glucagon rose to a greater extent during strenuous exercise and remained elevated after the end of exercise in the untrained group. Plasma human growth hormone rose to a greater extent during exercise and remained elevated after the end of exercise in the untrained group. Plasma cortisol fell at low and moderate exercise rates in both groups, but to a smaller extent in the cyclists. Cortisol values rose at higher workloads and were significantly higher in the cyclists at the end of exercise. 5. It is concluded that there are significant differences in the metabolic and hormonal responses to exercise between athletically trained and untrained individuals, even when the physically fit subjects work at the same percentage of their maximal capacity as the unfit subjects.


Highly Resolved Pure-Shift Spectra on a Compact NMR Spectrometer.
Benchtop NMR spectrometers experience a great success for a wide range of applications. However, their performance is highly limited by peak overlaps. Emerging "pure-shift NMR" (PS NMR) methods have been intensively used at high field to enhance the resolution by homodecoupling strategies. Here, different PS methods have been implemented on a compact NMR spectrometer operating at 43 MHz. Among the PS methods, the recent PSYCHE scheme appears more sensitive than Zangger-Sterk (ZS) experiments and offers a substantial resolution improvement as compared to 1D 1 H. On the other hand, despite their slightly lower sensitivity, ZS methods are more efficient to reduce broad signals and are more immune to strong couplings. Finally, the classical J-resolved pulse sequence is more efficient to reduce larger signals for bigger-sized molecules. The three approaches appear relevant for benchtop NMR and their combination forms an efficient toolbox to analyze a great diversity of samples.


An FTIR study of the induction and release of kiwifruit buds from dormancy.
Many deciduous, perennial fruit crops require winter chilling for adequate budbreak and flowering. Recent research has shown that changes in sugar and amino acid profiles are associated with the release of buds from dormancy. This paper uses FTIR spectrometry to provide an alternative mechanism for tracking metabolic changes in the meristems of kiwifruit buds during winter dormancy. The results suggest that the application of multivariate analysis to FTIR spectra has the potential to be a reliable and fast method for detecting structural and compositional changes in fruit crops. Ten wave numbers of the FTIR spectra are used to calculate a bud development function. This function has been validated using data from two seasons and four orchards, and by monitoring the effects of hydrogen cyanamide application, sugar concentrations and soil temperatures on this function. These wave numbers appear to be associated with carbohydrate, pectin and cellulose levels in the meristems. It is expected that this FTIR signature can be used to advance our understanding of the influence of the various environmental and physiological factors on the breaking of bud dormancy and shoot outgrowth, including the optimum timing and concentrations of applications of budbreak regulators, such as hydrogen cyanamide.


Improving the SAFT-γ Mie equation of state to account for functional group interactions in a structural (s-SAFT-γ Mie) framework: Linear and branched alkanes.
The group contribution SAFT-γ Mie EoS is based on the statistical associating fluid theory for fused heteronuclear molecules. While the chain term of the model has been modified to account for the new functional group-specific parameters, it does not impose a bonding order to these functional groups, only considering intergroup interactions in the monomer reference fluid. This leaves the model unable to account for the different physical properties of structural isomers and implicitly introducing modeling bias to species where the molecular structure mimics those used in the parameter regression. In this work, a simple but physically meaningful modification to the chain term in SAFT-γ Mie is proposed that accounts for the number of intergroup bonds, thereby encoding structural information in the model, without introducing an additional regressed parameter. The resulting structural SAFT-γ Mie (s-SAFT-γ Mie) requires reparameterization of the group parameters, which we present for linear and branched alkanes (CH3, CH2, CH, and C groups) here. Following an identical parameterization procedure to the original model, validation showed that the modification actually improves prediction accuracy for linear alkanes while addressing the original inability of the framework to distinguish between structural isomers. The good predictive performance seen in this work, for both pure component and mixture properties, lays a good foundation for expansion to other functional groups in future work.


Oncostatin M in William's E medium is suitable for initiation of hepatocyte differentiation in human induced pluripotent stem cells.
William's E (WE) is a suitable medium for the differentiation of human induced pluripotent stem (iPS) cells to the hepatocyte lineage. The aim of the present study was to investigate various growth factors in their ability to promote hepatocyte differentiation of iPS cells in WE medium. Human iPS 201B7 cells were cultured in WE medium supplemented with growth factors, and mRNA expression levels and promoter activities of α‑fetoprotein (AFP) and albumin were examined by reverse transcription‑quantitative polymerase chain reaction and luciferase assay, respectively. In addition, time course analysis of AFP mRNA expression was performed in 201B7 cells cultured in WE medium supplemented with oncostatin M. The results demonstrated that mRNA expression levels of AFP were significantly elevated by most growth factors tested as supplements in WE medium, except all‑trans retinoic acid, compared with cells cultured in ReproFF (a medium that maintains pluripotency). The highest increase in AFP mRNA expression levels was observed by oncostatin M stimulation. Albumin mRNA expression levels were increased by all‑trans retinoic acid and insulin‑transferrin‑selenium supplementation in WE medium compared with cells cultured in ReproFF. Oncostatin M supplementation significantly stimulated the promoter activity of the AFP gene, but no growth factor tested significantly stimulated the promoter activity of the albumin gene. By time course analysis, significant increase of AFP mRNA expression was observed on the sixth day post‑stimulation, compared with cells cultured in WE medium alone. In conclusion, the present study demonstrated that oncostatin M supplementation in WE medium was sufficient to initiate hepatocyte differentiation in iPS cells.


Synthesis and structure-activity relationships of ferrocenyl tamoxifen derivatives with modified side chains.
We report here the synthesis and cell-proliferation properties of derivatives of the breast cancer drug tamoxifen, in which the -O(CH(2))(2)N(CH(3))(2) side chain, responsible for the drug's antiestrogenic properties, has been modified by a ferrocenyl moiety. We recently reported the diphenol compound 5, in which this amino chain had been replaced with an acyl-ferrocenyl (-O(CH(2))(2)C(O)[(eta(5)-C(5)H(4))FeCp]) group, and which showed antiproliferative effects against both the hormone-dependent MCF-7 and -independent MDA-MB-231 breast cancer cell lines. We now report the results of a structure-activity relationship (SAR) study, in which the lateral chain length has been varied, the ketone group has been omitted, and the number of phenol groups has been varied. Compounds 1-4, with a side chain lacking the carbonyl function (-O(CH(2))(n)[(eta(5)-C(5)H(4))FeCp], n = 1-4) and which show a decreasing affinity for ERalpha (ER = estrogen receptor) with increasing chain length, act as estrogens on MCF-7 cells, and mild cytotoxics on PC-3 prostate cancer cells, with IC(50) values around 10 microM. The two monophenolic derivatives of 2, 2 a and 2 b, which show a reduced affinity for ERalpha compared to 2, are also estrogenic, but are only slightly cytotoxic. Finally, we have reexamined compound 5 and discovered that its antiproliferative effect against the MCF-7 cell line does not arise from antiestrogenicity as we had originally suspected, but by means of a cytotoxic pathway. This compound is also sensitive to the number of phenol groups as cell death is diminished when one of the hydroxyl groups is omitted (5 a and 5 b). Molecular modeling studies of the ligand-ERalpha binding stability are broadly consistent with the experimental binding affinity results for compounds 2, 2 a, 2 b, 5, 5 a, and 5 b. Electrochemical experiments show that 1-4, 2 a, and 2 b are stable to oxidation on the electrochemical timescale, unlike 5, 5 a, and 5 b, and that cytotoxicity is related to less positive phenol oxidation potentials. The SAR study shows that the presence of a ketone group and two phenol groups is necessary for strong receptor binding and cytotoxic effects, and that all compounds are estrogenic, despite the presence of a bulky side chain.


Respiratory changes in subclavian vein diameters predicts fluid responsiveness in intensive care patients: a pilot study.
The present pilot study investigated whether respiratory variation in subclavian vein (SCV) diameters correlates with fluid responsiveness in mechanically ventilated patients. Monocentric, prospective clinical study on fluid responsiveness in adult sedated, mechanically ventilated ICU patient, monitored with the PiCCO™ system (Pulsion Medical System, Germany), and requiring a fluid challenge (FC). A 10-min fluid bolus of 500 mL of 0.9% saline was administered. Cardiac output (CO) and dynamic parameters [stroke volume variation (SVV) and pulse pressure variation (PPV)] measured by transpulmonary thermodilution and pulse contour analysis (PiCCO™) as well as classical hemodynamic parameters were recorded at baseline and after FC. Fluid responsiveness was described as an increase in CO of ≥ 15%. Ultrasound measurements obtained in the subclavian long-axis view were used to calculate the SCVvariability index. A cut-off value for SCV variation for the prediction of fluid responsiveness was determined using receiver operating curve (ROC) analysis. Nine of 20 FCs (45%) induced an increase in CO of ≥ 15%. At baseline, the SCVvariability index was greater in responders than in non-responders (34.0 ± 21.4 vs. 9.0 ± 5.5; p = 0.0005). Diagnostic performance for the SCVvariability index revealed a cut-off value of 14 with a sensitivity of 100% [Confidence interval (CI) 95% (90; 100)] and a specificity of 82% [CI 95% (48; 98)] for the prediction of fluid responsiveness. Other parameters, such as SVV and PPV, could not predict fluid responsiveness. The correlation coefficient between CO variation and the SCVvariability index was 0.73 (p < 0.001). The SCVvariability index was a reliable, non-invasive parameter for the prediction of fluid responsiveness at the bedside of mechanically ventilated, critically ill patients in this pilot study.


Gram staining apparatus for space station applications.
A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space.


A cytomegalovirus protein with properties of herpes simplex virus ICP8: partial purification of the polypeptide and map position of the gene.
We demonstrated the presence of a single-stranded DNA-binding protein in human cytomegalovirus (CMV)-infected cells with properties analogous to those of herpes simplex virus (HSV) ICP8. Using monoclonal antibody specific for the CMV protein, we analyzed its fluorescence pattern and time of synthesis, mapped the gene encoding it by using a lambda gt11 library of CMV DNA fragments, and monitored its purification by phosphocellulose and DNA-Sepharose chromatography. In all characteristics we examined, the CMV protein behaved analogously to HSV ICP8. Our results are consistent with a functional role of CMV ICP8 in viral replication that is similar to that of HSV ICP8 and with the evolutionary conservation of the gene of interest in two divergent herpesviruses.


Intraoperative aortic dissection.
Intraoperative aortic dissection is a rare but fatal complication of open heart surgery. By recognizing the population at risk and by using a gentle operative technique in such patients, the surgeon can usually avoid iatrogenic injury to the aorta. Intraoperative transesophageal echocardiography and epiaortic scanning are invaluable for prompt diagnosis and determination of the extent of the injury. Prevention lies in the strict control of blood pressure during cannulation/decannulation, construction of proximal anastomosis, or in avoiding manipulation of the aorta in high-risk patients. Immediate repair using interposition graft or Dacron patch graft is warranted to reduce the high mortality associated with this complication.


Organizational and expressional uniqueness of a testis-specific mRNA transcript of protooncogene c-kit receptor in water buffalo Bubalus bubalis.
Protooncogene c-kit receptor is implicated with spermatogenesis, melanogenesis, and hematopoeisis, and undergoes tissue/stage specific alternate splicing. We have isolated 2973-bp full-length cDNA sequence (CDS) of this gene from testis and other tissues of water buffalo Bubalus bubalis. Upon comparison, the c-kit sequences showed tissue specific nucleotide changes resulting in novel truncated peptides. These peptides lacked intracellular and/or transmembrane domains in all the tissues except testis. Other alternately spliced tissue-specific transcripts were also detected, which are the integral parts of the open reading frame and have been reported in other mammals. Phylogenetic analysis of the sequences revealed unique tyrosine kinase domain in buffalo. Copy number calculation and expressional analysis of c-kit using real-time PCR established its single copy status and highest expression (137-177 folds) in testis compared to that (least) in liver. c-kit expression was detected in semen samples although 10 times lesser compared to that in testis. The highest expression of c-kit in testis and the presence of mRNA transcript in sperms substantiate its predominant role in spermatogenesis. This study establishes unequivocal involvement of an autosomal gene c-kit receptor in testicular function.


Pain experienced by electric-powered chair users: a pilot exploration using pain drawings.
Many wheelchair users experience pain or discomfort, and little is known about its severity or nature. The present study explored the feasibility of using visual analogue scales and pain drawings to assess the intensity and distribution of pain in users of electric-powered indoor and outdoor wheelchairs, and its changes over time. Five electric-powered indoor and outdoor chair (EPIOC) users, mean age 62 years (range 43-84 years) completed pain drawings and visual analogue scales two-hourly during one day. Pain intensity increased throughout the day in all users. It was most severe in the neck, back and buttocks. Pain drawings gave visual impact to the extent of the pain. Visual analogue scales and pain drawings may be useful in assessing pain experienced by EPIOC users, which may be severe and appears to increase in severity and distribution during the day.


Polishing the tarnished silver bullet: the quest for new antibiotics.
We are facing a potential catastrophe of untreatable bacterial infections, driven by the inexorable rise of extensively drug-resistant bacteria, coupled with a market failure of pharmaceutical and biotech companies to deliver new therapeutic options. While global recognition of the problem is finally apparent, solutions are still a long way from being implemented. In addition to drug stewardship programmes and better diagnostics, new antibiotics are desperately needed. The question remains as to how to achieve this goal. This review will examine the different strategies being applied to discover new antibiotics.


Lead levels in paint flakes from buildings in Nigeria: a preliminary study.
Lead is a malleable metal previously used to improve the durability and color luster of paint applied in homes and on industrial structures such as bridges. Lead has deleterious effects on multiple organs in humans. There is paucity of information on the extent of the use of lead-based paint in Nigerian houses. This study has attempted to estimate the extent of use of lead-based paint in buildings in Eastern Nigeria using 168 buildings. Flaked paint samples were collected from residential, church, commercial, and school buildings from four most populous cities in Eastern Nigeria namely Enugu, Onitsha, Aba, and Port Harcourt, and they were digested using conc HNO(3):HCLO(4) (1:1) and analyzed using atomic absorption spectrophotometer. The lead levels from buildings in Enugu ranged from 39.385 +/- 1.111-69.843 +/- 4.886 mg/kg. The highest level of lead was found in Onitsha ranging from 49.503 +/- 0.000-74.352 +/- 0.571 mg/kg. Residential buildings, which mainly serve the under privileged populations, has the highest lead level in this study. In Aba, the highest lead level (66.432 +/- 0.013 mg/kg) was found in commercial buildings aged 5-10 years. The lead levels in paint flakes from buildings in the four cities tended to decrease with increasing age of the buildings. Taken together all the building paint flakes from the four cities had lead levels higher than the United States Environmental Protection Agency permissible level of 5 mg/kg. There is a need for primary intervention strategy to reduce the paint lead levels in Nigeria.


Fe2TiO5/Fe2O3 (Shell/Shell) and (Shell/Core) Heterostructured for Efficient Oxygen Evolution.
A delicate synthesis strategy of Fe2TiO5/Fe2O3 (shell/shell (s/s)) and (shell/core (s/c)) heterostructures was proposed to realize efficient photocatalytic water oxidation. The hierarchical structure increases the light absorption depth of material, the strongly coupled components promoted the separation of photoinduced excitons, and the high surface area with adequate porosity allowed the diffusion of electrolyte to adsorb at rich active sites. Furthermore, Fe2TiO5/Fe2O3 (s/s) was coated by graphitic carbon nitride (g-CN) subunits and exhibited good photocatalytic water reduction, resulting from the contact interface accelerating the effective separation of electrons and holes.


Testicular transposition in children before scrotal external radiotherapy.
Testicular transposition (TT) before scrotal external radiotherapy (RT) is poorly reported in children with cancer, with only rare case reports published. TT surgical techniques, dosimetric parameters, and testicular functions are retrospectively reported in 12 children, median age 12.8 years, after scrotal RT for sarcomas. TT has low morbidity and allows a dramatic RT dose decrease in the healthy testicle. Endocrine functions seem preserved while more follow-up is needed to assess fertility. Though a rare situation, TT should be discussed in children and young adult cases when a scrotal high-dose RT is needed.


[Screening for thyroid disorders in elderly patients].
In elderly patients, thyroid diseases may remain undiagnosed due to the lack of specificity of the clinical presentation. Thyroid function alterations seem to be more common in older persons than in adults. The aim of our study was to evaluate the incidence of thyroid function alterations in 300 elderly patients admitted to the division of Internal Medicine of the local hospital in Levanto, in a one-year period. Thyroid function alterations were discovered in 12.6% of the patients and considering the group of patients in whom a thyroid function alteration was demonstrated, 45% of them were affected by hypothyroidism (10.7% overt primary hypothyroidism, 28.9% sub-clinical hypothyroidism, 5.4% hypothyroidism secondary to hypopituitarism), 15.6% by hyperthyroidism (overt 7.8% , subclinical 7.8%), and 39.4% showed a low T3 syndrome. Our data confirm the high incidence of previously unrecognized thyroid diseases in the elderly patients admitted to an hospital and the profit that these patients can receive from the appropriate diagnosis at the admission and justify the cost of routine testing for FT4 and TSH in every person at hospital admission.


Nonspecificity of intraparenchymal blood-fluid levels in cerebral computed tomography.
In previous reports statements have been made that blood-fluid levels seen on cerebral CT were indicative of preexisting tumor or arteriovenous malformations. Three cases are presented which indicate that an intraparenchymal blood-fluid level is a nonspecific finding that may be encountered in other lesions than those previously reported.


Type-II external fixation, using new clamps and positive-profile threaded pins, for treatment of fractures of the radius and tibia in dogs.
A new external fixation system for repair of fractures of the radius and tibia was developed that uses positive-profile threaded pins. This system allows for addition of a fixation clamp between 2 installed clamps, predrilling of pin holes through a drill sleeve, use of positive-profile threaded pins in all locations, and easier application of full pins. Type-II external fixators were applied, using this system, to fractures in 10 client-owned dogs, and outcome was evaluated. All fractures healed without complications. Duration of surgery, mean time until radiographic evidence of a bridging callus, and mean time until removal of the external fixator were shorter, and frequency of pin loosening were less than with other techniques. Pin loosening was uncommon. This system provides an important improvement in external skeletal fixation.


Associations of NR5A2 gene polymorphisms with the clinicopathological characteristics and survival of gastric cancer.
The orphan nuclear receptor (NR5A2), which belongs to the NR5A subfamily of nuclear receptors, is expressed in developing and adult tissues of endodermal origin, and can contribute to the development of several cancers through regulating cell proliferation. NR5A2 (rs3790843 and rs3790844) single nucleotide polymorphisms (SNPs) genotyping were examined in DNA samples, extracted from paraffin-embedded cancer tissue. Clinicopathologic and follow-up data were collected from 944 patients with gastric cancer (GC). Associations of the 2 SNPs with the progression and prognosis in gastric cancer patients were analyzed using the SPSS version 18.0. We found that NR5A2 rs3790843 polymorphism was significantly associated with the risk of GC which had regional lymph node metastasis (p = 0.044) or distant metastasis (p = 0.020). Our results also indicated that rs3790844 polymorphism was associated with the increased overall survival (OS) of GC patients in the dominant model (GG vs. GA/AA, HR (hazard ratio) = 0.823, 95% CI (confidence interval) = 0.679-0.997), suggesting a potential protective role of the variant A allele. Additionally, in the stratified analysis, both NR5A2 rs3790843 and rs3790844 polymorphism were associated with significantly lower risk of death in the groups of female, tumor size >5 cm in a dominant model. Our results represent the first demonstration that the NR5A2 rs3790844 polymorphism is associated with increased OS of GC patients in the dominant model, and similar results were found among the female group and tumor size >5 cm group for NR5A2 rs3790843 polymorphism. Further validation in other larger studies with different ethnic populations and functional evaluations are needed.


Multi-level regulation of coelimycin synthesis in Streptomyces coelicolor A3(2).
Despite being a yellow pigment visible to the human eye, coelimycin (CPK) remained to be an undiscovered secondary metabolite for over 50 years of Streptomyces research. Although the function of this polyketide is still unclear, we now know that its "cryptic" nature is attributed to a very complex and precise mechanism of cpk gene cluster regulation in the model actinomycete S. coelicolor A3(2). It responds to the stringent culture density and timing of the transition phase by the quorum-sensing butanolide system and to the specific nutrient availability/uptake signals mediated by the global (pleiotropic) regulators; many of which are two-component signal transduction systems. The final effectors of this regulation cascade are predicted to be two cluster-situated Streptomyces antibiotic regulatory proteins (SARPs) putatively activating the expression of type I polyketide synthase (PKS I) genes. After its synthesis, unstable, colorless antibiotic coelimycin A reacts with specific compounds in the medium losing its antibacterial properties and giving rise to yellow coelimycins P1 and P2. Here we review the current knowledge on coelimycin synthesis regulation in Streptomyces coelicolor A3(2). We focus on the regulatory feedback loop which interconnects the butanolide system with other cpk cluster-situated regulators. We also present the effects exerted on cpk genes expression by the global, pleiotropic regulators, and the regulatory connections between cpk and other biosynthetic gene clusters.


Analysis of medication off-odors using an electronic nose.
Packaging materials have been implicated as a source for off-odors in pharmaceutical products. A new instrumentation method employing an array of conducting polymer gas sensors was used to identify the offending packaging components in the canister of a pharmaceutical inhalant. A case study is described in which tainted inhalers as well as elastomeric components of the canisters were 'sniffed' by the electronic nose. The electronic nose was able to differentiate between tainted and untainted canisters. Signal processing algorithms performed on the raw data from the sensors suggested that specific elastomeric components were responsible for the off-odor. A further experiment suggested that the propellant (Freon) extracted the odor from the elastomeric components as the medication was expelled from the canister. These data indicate that the electronic nose is a potential tool to solve odor problems in which human odor assessment is not feasible due to excess exposure to the medically active ingredient.


Release of leukotrienes during rapid expulsion of Trichinella spiralis from immune rats.
Rapid expulsion of the nematode Trichinella spiralis from immune rats is associated with an increase in volume of intestinal exudate and the presence of large numbers of tissue mucosal mast cells (MMC) and eosinophils. We have measured the concentrations of leukotrienes (LT) C4 (LTC4) and B4 (LTB4) in gut perfusates and mucosal homogenates at 30 min, 1, 3, 6 and 20 hr after challenge with larvae. Leukotrienes were identified by radioimmunoassay (RIA) combined with reverse-phase high-pressure liquid chromatography (RP-HPLC). There were significant elevations at 30 min and 1 hr in the concentrations of LTC4 in the perfusates from the gut of challenged immune rats compared to controls (infected unchallenged and uninfected naive rats). Similar increases in immunoreactive LTC4 and LTB4 were observed in mucosal homogenates from the gut of immune challenged animals. A second peak of LTB4 was also observed at 20 hr in both immune and naive challenged rats. There were also elevations in serum concentration of the MMC-associated specific serine protease, rat mast cell protease II (RMCPII). Since LTC4 causes smooth muscle contraction, increased vascular permeability and stimulation of mucus hypersecretion, and LTB4 recruits and activates inflammatory cells, leukotrienes may participate in the process of rapid expulsion of T. spiralis.


Mechanism of interactions between soyasaponins and soybean 7S/11S proteins.
In this study, the proteins glycinin (11S) and β-conglycinin (7S) were mixed with soyasaponin (Ssa) Ab/Bb to form a composite system. We used fluorescence and synchronous fluorescence spectra to demonstrate the changes in the surrounding environment and the structure of the proteins. Dynamic interface behavior analysis showed the possible interface behavior induced by the composite system. The interactions between Ssa and the proteins, along with the mode of action, were analyzed by molecular docking. The interactions between Ssa and soy protein increased with the change in concentration. The interactions between the two proteins were mediated by tryptophan (Trp) and primarily involved hydrogen bonds, which changed the microenvironment and loosened the protein structure. These results helped in understanding the mechanism underlying the interactions between Ssa Ab/Bb and 7S/11S. Furthermore, these results highlighted the theoretical fundamentals for the future applications of composite systems as surfactants in the food industry.


A case study using a patient satisfaction survey to improve the delivery and effectiveness of drug addiction treatment services: marketing implications and organizational impact.
Drug abuse and addiction continues to negatively impact many lives in this country. The United States health care system has grappled with how to best serve this vulnerable population. Since the personal and societal costs of addiction are high, all recent iterations of the United States strategic health plans (such as Healthy People 2010) have prioritized this area for improvement. At the local level, health care providers who care for those with addictions are challenged with shrinking insurance coverage for services, a difficult patient population, lack of treatment options, growing ranks of indigent patients, as well as a plethora of additional management challenges. It is known that successful treatment is integrally linked with patient satisfaction with services. The most critical factors in successful addiction treatment (from a patient's perspective) are (1) their belief that the counselor cares about them and, (2) their belief that they can recover. This paper reports a case study in the use of a patient satisfaction survey as a quality management/service refinement tool within a methadone treatment setting. Results indicate that the use of the survey itself provides patients with a tangible cue supporting the presence of the critical success factors. Further, the use of a survey provides a baseline for future measurements and trending. The paper concludes with a discussion of the marketing and organizational implications of incorporating the patient satisfaction survey into the ongoing delivery program for addiction services.


Interaction of Human OATP1B1 with PDZK1 Is Required for Its Trafficking to the Hepatocyte Plasma Membrane.
Uptake of xenobiotics by hepatocytes is mediated by specific proteins, including organic anion transporting polypeptides (OATPs), residing on the basolateral (sinusoidal) plasma membrane. Many of the OATPs have PDZ consensus binding sites, determined by their C-terminal 4 amino acids, while others do not. Mouse and rat OATP1A1 are associated with PDZK1, which is necessary for their trafficking to the plasma membrane. humanOATP1B1 (hOATP1B1) is a major drug transporter in human liver. Although localized to the plasma membrane, it was thought to lack a PDZ consensus motif, suggesting that the trafficking paradigm for murine OATPs is not applicable to human liver. The aim of the present study was to determine whether hOATP1B1 is a ligand for hPDZK1. hOATP1B1 immunoprecipitates with hPDZK1 following co-expression in 293T cells as well as in normal human liver. Co-expression with each of the 4 PDZ domains revealed interaction with domain 1 only. A truncated version of hOATP1B1 that lacks its terminal 4 amino acid PDZ binding motif as well as hOATP1B3, which does not contain a PDZ binding consensus motif, failed to interact with hPDZK1. Immunofluorescence microscopy of hOATP1B1 in stably transfected HeLa cells that endogenously express hPDZK1 showed that it distributes predominantly along the plasma membrane whereas hOATP1B1 lacking its terminal 4 amino acids distributes primarily intracellularly with little plasma membrane localization. Similar to findings in rats and mice, human OATP1B1 is a ligand for PDZK1 and requires interaction with PDZK1 for optimal trafficking to the hepatocyte plasma membrane. SIGNIFICANCE: Previous studies suggested that OATP1B1, a major xenobiotic transporter in human liver, does not have a PDZ binding consensus motif and does not follow the paradigm for subcellular trafficking and function that was established for OATP1A1 in murine liver. We now demonstrated that OATP1B1 but not OATP1B3 has a PDZ binding consensus motif that mediates binding to PDZK1 and is required for its trafficking to the plasma membrane. Such interaction could be an important previously unrecognized modulator of transport function.


Acute-phase blood pressure trajectories and clinical outcomes in ischemic stroke.
High blood pressure (BP) is frequent in acute ischemic stroke (IS). However, the impact of BP change patterns during acute phase on clinical outcomes is not conclusive. This study aims to investigate the association between the acute-phase BP trajectories and clinical outcomes in IS patients with high admission BP. The cohort consisted of 316 IS patients with admission systolic BP (SBP) ≥160 mm Hg. SBP trajectories during the first 7 days after onset were characterized using a random effects model. The patients were classified into three groups based on the SBP trajectory curve parameters: sustained high SBP (T1), moderate decrease (T2), and rapid decrease in SBP (T3). Poor outcomes were defined as modified Rankin scale score ≥3 in 3 months after onset. The relationship between SBP trajectory groups and the outcome was examined in multivariable logistic regression models. The decreasing trend was greater in the favorable than in the poor outcome group (P = 0.028 for difference in linear slopes). The incidence of poor outcomes was 25.9%, 13.5%, and 9.8% in T1 (n = 54), T2 (n = 170), and T3 (n = 92) groups, respectively. Compared with T1 group, the decrease in SBP in T2 and T3 groups was significantly associated with lower risk of the poor outcome (odds ratio = 0.25, 95% confidence interval = 0.10-0.67, P = 0.006). These findings suggest that a decrease in BP in the acute phase is predictive of favorable outcomes in IS patients. BP trajectories have a greater power to detect the association than individual BP values at one time-point.


Effect of the 2014 Clinical and Laboratory Standards Institute urine-specific breakpoints on cefazolin susceptibility rates at a community teaching hospital.
Enterobacteriaceae, which include Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis, are identified as the infectious etiology in the majority of urinary tract infections (UTIs) in community hospitals across the United States. The minimum inhibitory concentration (MIC) is a useful tool when choosing an appropriate antibacterial agent. Recent changes to the 2014 Clinical and Laboratory Standards Institute (CLSI) guidelines included reporting a urine-specific cefazolin breakpoint for enterobacteriaceae (susceptible ≤16 mcg/mL). The purpose of this study was to determine the clinical and financial impact of implementing the 2014 CLSI urine-specific breakpoints for cefazolin in a community-based teaching hospital in the Southern U.S.A. A retrospective review of patients hospitalized from January 1, 2010 through October 1, 2014 was performed. Patients that met inclusion criteria had a documented initial clinical isolate of E. coli, K. pneumoniae, or P. mirabilis from urine cultures during each year. Descriptive statistics and two-proportion test of hypothesis were used in the analysis to compare susceptibility rates before and after implementation of the updated CLSI breakpoints for cefazolin. A total of 190 clinical isolates from patients were included in the study. E. coli was the most common organism isolated (63.7%), followed by K. pneumoniae (22.1%), and P. mirabilis (14.2%). 86% of the included isolates were susceptible to cefazolin using the 2010 breakpoints. Implementation of the 2014 breakpoints did not significantly impact susceptibility results for E. coli, K. pneumoniae, or P. mirabilis. Modification of breakpoints did not significantly impact susceptibility rates of cefazolin. Substituting cefazolin may decrease the overall drug cost by 77.5%. More data is needed to correlate in vitro findings with clinical outcomes using cefazolin for UTIs.


Modulation of HIV replication in monocyte derived macrophages (MDM) by steroid hormones.
Significant sex specific differences in the progression of HIV/AIDS have been reported. Several studies have implicated steroid hormones in regulating host factor expression and modulating HIV transmission and replication. However, the exact mechanism exerted by steroid hormones estrogen and progesterone in the regulation of HIV-1 replication is still unclear. Results from the current study indicated a dose dependent down regulation of HIV-1 replication in monocyte derived macrophages pre-treated with high concentrations of estrogen or progesterone. To elucidate the molecular mechanisms associated with the down regulation of HIV-1 replication by estrogen and progesterone we used PCR arrays to analyze the expression profile of host genes involved in antiviral responses. Several chemokines, cytokines, transcription factors, interferon stimulated genes and genes involved in type-1 interferon signaling were down regulated in cells infected with HIV-1 pre-treated with high concentrations of estrogen or progesterone compared to untreated HIV-1 infected cells or HIV-1 infected cells treated with low concentrations of estrogen or progesterone. The down regulation of CXCL9, CXCL10 and CXCL11 chemokines and IL-1β, IL-6 cytokines in response to high concentrations of estrogen and progesterone pre-treatment in HIV-1 infected cells was confirmed at the protein level by quantitating chemokine and cytokine concentrations in the culture supernatant. These results demonstrate that a potent anti-inflammatory response is mediated by pre-treatment with high concentrations of estrogen and progesterone. Thus, our study suggests a strong correlation between the down-modulation of anti-viral and pro-inflammatory responses mediated by estrogen and progesterone pre-treatment and the down regulation of HIV-1 replication. These findings may be relevant to clinical observations of sex specific differences in patient populations and point to the need for further investigation.


Understanding the Effects of Smart-Speaker-Based Surveys on Panelist Experience in Immersive Consumer Testing.
Utilizing immersive technologies to reintroduce the environmental context (i.e., visual, auditory, and olfactory cues) in sensory testing has been one area of research for improving panelist engagement. The current study sought to understand whether pairing smart-speaker questionnaires in immersive spaces could positively affect the panelist experience through enhanced ecological validity. To this end, subjects performed an immersive consumer test in which responses were collected using a traditional computer-based survey, a smart-speaker approach incorporating a direct translation of the computer questionnaire into a verbal survey requiring numeric responses, and an optimized smart-speaker survey with alternative question formatting requiring spoken word-based responses. After testing, participants answered the Engagement Questionnaire (EQ) to assess participant engagement during the test, and the System Usability Scale (SUS) survey to understand the ease, and potential adoption, of using the various survey technologies in the study. Results indicated that the traditional computer-based survey was the most engaging (p < 0.001) and usable (p < 0.001), with no differences found between the two smart-speaker surveys (p = 0.803 and p = 0.577, respectively). This suggests that the proposed optimizations for the smart-speaker surveys were not robust enough to influence engagement and usability, and further research is needed to enhance their conversational capabilities.


AX+, BX- discrimination learning in the fear-potentiated startle paradigm: possible relevance to inhibitory fear learning in extinction.
The neural mechanisms of fear suppression most commonly are studied through the use of extinction, a behavioral procedure in which a feared stimulus (i.e., one previously paired with shock) is nonreinforced repeatedly, leading to a reduction or elimination of the fear response. Although extinction is perhaps the most convenient index of fear inhibition, a great deal of behavioral work suggests that postextinction training conditioned stimuli are both excitatory and inhibitory, making it difficult to determine whether a neural manipulation affects inhibition, excitation, or some combination thereof. For this reason we sought to develop a behavioral procedure that would render a stimulus primarily inhibitory while at the same time avoiding some of the issues raised by the traditional conditioned inhibition paradigm, namely second-order conditioning, external inhibition, and configural learning. Using the fear-potentiated startle paradigm, we adapted an AX+, BX- training procedure in which stimuli A and X were presented simultaneously and paired with shock, and stimuli B and X were presented simultaneously in the absence of shock. In testing, high levels of fear-potentiated startle were seen in the presence of A and AX and much lower levels were seen in the presence of B and AB, as would be predicted if stimulus B were a conditioned inhibitor. We believe this method is a viable alternative to the traditional conditioned inhibition training procedure and will be useful for studying the neural mechanisms of fear inhibition.


Identifying atomic geometry and electronic structure of (2 x 3)-Sr/Si(100) surface and its initial oxidation.
We present a joint experimental and theoretical study on the geometric and electronic states and the initial oxidation of the (2x3)-Sr/Si(100) surface. With scanning tunneling microscopy/scanning tunneling spectroscopy (STM/STS) measurements combined with ab initio calculations, the atomic geometry and the electronic states of the (2x3)-Sr/Si(100) surface are identified. The dimerization of the Si atoms in the single atom row based on a (1x3) Si substrate model plays a critical role in stabilization of the surface structure and in determining the electronic properties. At the very initial oxidation of the surface, four features corresponding to the primary adsorption and oxidation sites are determined. Three of them are corresponding to the most favored oxidation sites with single oxygen molecules, whose local density of states gives semiconducting behavior. One is corresponding to the oxidation site with two oxygen molecules, whose local density of states gives metallic behavior. These features all exhibit dark spots with different shapes in the occupied state images but display either dark spots or bright protrusions depending on the different oxidation sites in the empty state images. Compared with the theoretical calculations, the plausible adsorption and oxidation models are proposed.


Familial gastrointestinal stromal tumor syndrome: report of 2 cases with KIT exon 11 mutation.
As with cases of sporadic gastrointestinal stromal tumor (GIST), familial GIST syndrome arises from mutations in KIT or PDGFRA. Only a few dozen such families have been described in the literature. Cases of 2 individuals from 2 different newly described kindreds with familial GIST syndrome were retrospectively reviewed. Pertinent immunohistochemical stains, including CD117, CD34, DOG1, desmin, and S100, were performed. Samples from each case were sent to outside facilities for molecular analysis. A review of the relevant literature was performed and the number of familial GIST syndrome cases reported was updated through July 2014. In case 1, a woman 40 years of age with a family history of GIST presented with abdominal pain and gastrointestinal bleeding. Biopsy of a gastric mass revealed spindle-cell type GIST. Molecular analysis revealed a heterozygous mutation of p.Asp579del in exon 11 of KIT. The patient was placed on imatinib therapy and an initial positive response was demonstrated by imaging. Disease regression was seen on computed tomography, and several GIST tumors were surgically resected. The patient has had stable disease since surgery. In case 2, an asymptomatic woman 29 years of age presented for screening due to a family history of GIST. One small nodule was noted in her stomach and another was noted in the duodenum; both were surgically resected. The patient recovered well following surgery. The GIST in this patient was noted to have similar histological, immunohistochemical, and molecular findings as case 1. Imatinib has often been shown to be an effective therapy in both the familial and sporadic forms of GIST. There is no standard protocol for addressing the surveillance of patients with spindle-cell type GIST seen in the setting of familial GIST syndrome and with a p.Asp579del mutation of exon 11 on KIT.


Effect of interleukin-2 on salivary gland morphology and function in mice.
C57BL mice were injected intraperitoneally with 300,000 Cetus units/day of human recombinant interleukin-2 (rIL-2) for 2, 4, and 5 days to study its effect on salivary gland function and morphology. The pilocarpine-stimulated parotid salivary flow was collected via cannulation of the glandular duct. Total salivary protein was assayed spectrophotometrically, salivary electrolytes were determined by atomic absorption, and glandular lymphoid cell infiltration was evaluated histologically. After 5 days of rIL-2 administration salivary output and total salivary protein concentrations were reduced significantly. Similar changes, albeit to a lesser extent, were observed following 2 and 4 days of rIL-2 treatment. Increased lymphoid infiltration of the salivary glands was observed, and was directly related to the duration of rIL-2 administration. The effect of the lymphokine on the parotid gland gradually dwindled after cessation of treatment: 4 days posttreatment this salivary gland showed signs of recovery, which at 10 days proved to be complete. The possible use of this animal model in the study of lymphocyte-induced salivary gland diseases is discussed.


Adenosine 5'-triphosphate synthesis in Escherichia coli submitted to a microsecond electric pulse.
The total cytoplasmic ATP content (bound and free) increased in Escherichia coli when the bacteria were submitted to electric pulses with field strengths of 1-6 kV/cm and a decay time of 7-20 microseconds. The electron-transport chain was blocked by cyanide, and ATP synthesis was detected by a luminescence assay. The amount of newly formed ATP depends on the field strength. A total of 150 pmol of ATP was formed per milligram of bacteria submitted to a 3 kV/cm pulse. Synthesis was blocked by uncouplers and ionophores (valinomycin). The F1F0-ATP synthase inhibitor dicyclohexylcarbodiimide blocked a large part of this synthesis. Synthesis was not induced in unc mutants (unc B, unc D). The synthesis of ATP is related to the induced transmembrane potential, not to the Joule heating. A minimum 35-50-mV increase in membrane potential must be maintained for at least 12 microseconds to trigger this synthesis. This very fast energy transduction in bacteria is in good agreement with our previous results concerning submitochondrial particles. Because of the localized character of the induced membrane potential, these results are in agreement with the recent hypothesis of "mosaic proton coupling".


Staining of surface antigen on migrating lymphocytes in the chemotaxis membrane. I: Complement C5 attracts helper/inducer T cell much more than suppressor/cytotoxic T cell and NK cell.
We have measured various chemotactic activities for neutrophils (1-3), lymphocytes (4), macrophages (5) and eosinophils (6-11) by the modified Boyden chamber method. In the conventional method, it is necessary to purify the indicator cells in upper chamber because it is impossible to distinguish inflammatory cells on the chemotactic membrane. For example, when we estimate T lymphocyte chemotaxis, we collected nylon-wool nonadherent peripheral blood lymphocytes (PBL) and confirmed their purity by sheep erythrocytes rosette formation assay. Pohajdak et al (12) evaluated NK cell chemotaxis by using large granular lymphocytes (LGL) for indicator cells. LGL fractionation was confirmed by monoclonal antibodies, HNK-1, OKT11, OKT3, OKT4, OKT8, OKM1, and MO2. When we arranged the study to evaluate chemotaxis of lymphocyte subsets which were divided by surface antigens, the conventional method needed a large quantity of PBL for the indicator cells. Moreover, one or two lymphocyte subsets had to be obtained from an identical PBL donor. This was time consuming. So, we tried to identify migrating lymphocytes in the chemotaxis membrane by their surface antigens. In this communication, we report that the staining of surface antigens on migrating lymphocytes in the chemotaxis membrane is effective and useful for evaluation of lymphocyte subset chemotaxis. It prompted us to report the new findings about modification of lymphocyte subset chemotaxis by C5a.


The role of motility as a virulence factor in bacteria.
Many bacteria that cause diseases of humans, animals and plants use flagella to move. This review summarises recent studies that have analysed the role of motility and chemotaxis in the host-parasite relationship of pathogenic bacteria. These studies have shown that for many pathogens, motility is essential in some phases of their life cycle and that virulence and motility are often intimately linked by complex regulatory networks. Possibilities to exploit bacterial motility as a specific therapeutic antibacterial target to cure or prevent disease are discussed.


Experimental Chua-plasma phase synchronization of chaos.
Experimental phase synchronization of chaos is demonstrated for two different chaotic oscillators: a plasma discharge and the Chua circuit. Our technique includes real-time capability for observing synchronization-desynchronization transitions. This capability results from a strong combination of synchronization and control, and allows tuning adjustments for search and stabilization of synchronous states. A power law is observed for the mean time between 2pi phase slips for different coupling strenghts. The experimental results are consistent with the numerical simulations.


Non-plastidic, tyrosine-insensitive prephenate dehydrogenases from legumes.
L-Tyrosine (Tyr) and its plant-derived natural products are essential in both plants and humans. In plants, Tyr is generally assumed to be synthesized in the plastids via arogenate dehydrogenase (TyrA(a), also known also ADH), which is strictly inhibited by L-Tyr. Using phylogenetic and expression analyses, together with recombinant enzyme and endogenous activity assays, we identified prephenate dehydrogenases (TyrA(p)s, also known as PDHs) from two legumes, Glycine max (soybean) and Medicago truncatula. The identified PDHs were phylogenetically distinct from canonical plant ADH enzymes, preferred prephenate to arogenate substrate, localized outside of the plastids and were not inhibited by L-Tyr. The results provide molecular evidence for the diversification of primary metabolic Tyr pathway via an alternative cytosolic PDH pathway in plants.


Nonlinear electrical impedance spectroscopy of viruses using very high electric fields created by nanogap electrodes.
Our living sphere is constantly exposed to a wide range of pathogenic viruses, which can be either known, or of novel origin. Currently, there is no methodology for continuously monitoring the environment for viruses in general, much less a methodology that allows the rapid and sensitive identification of a wide variety of viruses responsible for communicable diseases. Traditional approaches, based on PCR and immunodetection systems, only detect known or specifically targeted viruses. We here describe a simple device that can potentially detect any virus between nanogap electrodes using nonlinear impedance spectroscopy. Three test viruses, differing in shape and size, were used to demonstrate the general applicability of this approach: baculovirus, tobacco mosaic virus (TMV), and influenza virus. We show that each of the virus types responded differently in the nanogap to changes in the electric field strength, and the impedance of the virus solutions differed depending both on virus type and virus concentration. These preliminary results show that the three virus types can be distinguished and their approximate concentrations determined. Although further studies are required, the proposed nonlinear impedance spectroscopy method may achieve a sensitivity comparable to that of more traditional, but less versatile, virus detection systems.


Characterization of the Paracoccidioides beta-1,3-glucanosyltransferase family.
The cell wall of pathogenic microbes acts as an initial barrier that is in contact with hostile environments. Several proteins are associated to the cell wall, including the glucanosyltransferases, which are attached through glycosylphosphatidylinositol anchors to the wall. Here, we characterized the Paracoccidioides beta-1,3-glucanosyltranferase ( Gel ) family of proteins that showed significant homology to proteins belonging to the GH72 family. Immunoassays demonstrated Gel1p associated with the cell wall and with the nucleus. For Gel2p, immune labeling was associated with the cell wall and cytoplasm. Genetic complementation studies in Saccharomyces cerevisiae demonstrated that Gel2p is able to participate in the maintenance of fungal cell wall integrity, as it was able to restore the lack of Gas1p activity in a gas1Δ mutant; Gel1p was not able to do the same. On the other hand, Gel1p restores telomeric silencing in a gas1Δ mutant, providing strong support that Gel1p can be involved in transcriptional silencing in Paracoccidioides. Use of the in vivo yeast two-hybrid system revealed proteins that interact with Paracoccidioides Gel proteins, supporting new insights into the function of Gel family members and suggesting that they could play other roles than those established at the fungal cell wall.


Autoantibodies to types I and IV collagen and heart valve disease in systemic lupus erythematosus/antiphospholipid syndrome.
Introduction/objectives autoantibodies to types I and IV collagen have been described in rheumatic fever and infective endocarditis. We tried to elucidate if an autoimmune response against collagens I and IV exists, associated with heart valve disease in systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS). A cohort of 172 patients with SLE (n = 79), primary APS (PAPS, n = 83), and secondary APS (n = 10) were assessed for valvulopathy by transthoracic echocardiograms. Autoantibodies to types I and IV collagen were assessed in patients and 50 controls, setting autoantibody positivity at two standard deviations above the mean antibody level of controls. Positive anticollagen IV antibody rate was significantly higher in SLE patients (17.7%) in respect to the rest of groups (PAPS 2.4%, controls 2%; P = 0.001). Percentage of positive autoantibodies to collagen I was similar in SLE and APS cohort of patients with and without valvular disease (48.4 vs 51.6%, respectively; P = 0.45). Percentage of positive autoantibodies to collagen IV was increased but not significantly in SLE and APS cohort of patients with respect to those without valvular disease (62.5 vs 37.5%, respectively; P = 0.08). Mean (standard deviation) levels of positive anticollagen I and IV antibodies did not differ between patients with and without valvular disease (85.6 ± 55 vs 81 ± 85 U/ml, respectively; P = 0.86 for anticollagen I) (0.05 ± 0.02 vs 0.12 ± 0.16 U/ml, respectively; P = 0.34 for anticollagen IV). Our data indicate a lack of association of autoantibodies to types I and IV collagen with heart valve disease in SLE and APS.


When the spirit leaves: Childhood death, grieving, and bereavement in Islam.
The death of a child has a profound and often long-lasting impact on families. The parent's relationship and their ability to bond with and take care of surviving children may be affected. It is important for healthcare workers to understand the dynamics associated with bereavement, especially when the family comes from a non-Western culture. Islam is one of the three most populous religions along with Christianity and Hinduism and the fastest growing religion in the United States but remains largely misunderstood. This paper seeks to explain what Islam is, who is a Muslim, where they live, and what they believe and practice. It also explains how Islamic beliefs contextualize the meaning of life and death for Muslims and how they are exhorted to grieve upon a child's death. Reading this paper will enable those who care for Muslim families to better attend to the social and emotional needs of Muslim parents and siblings after such a tragic event.


Medicine, old age and public interest in historical perspective. The German case.
This paper outlines the history of social disinterest on old people from the perspective of medical discourse since the end of the 18th century. The following thesis is proposed: since the Enlightenment medical discourse over illnesses associated with old age has been masked by the social valuation that dying is attributable to old age and old people could not make any great demands on medical assistance. Until the end of the 19th century it was theologians, not doctors, who were responsible for old people on the threshold to death. Around 1900 great interest arose in the problem complex "aging" and "rejuvenation" from the view point of bio-chemistry. But the associated cultural circumstances, namely a dominant social interest in youth and fitness, largely prevented an original medical interest in old people. This has only developed since the 1950s, not within the medical profession but in the "social" professions, primarily social work. From this perspective the application of Foucault's paradigm, in which old age, actually a social problem, is defined and addressed by medicine, in other words a "medicalization of old age", seems to be only conditionally meaningful as a conceptual framework for analysis.


Treatment of congestive heart failure. Pathophysiologic and pharmacologic concepts.
Congestive heart failure is a serious health problem in Western society. Understanding of both the basic pathophysiology and the design of rational and effective treatment of this syndrome has increased dramatically in the last ten years. However, congestive heart failure remains a highly lethal disease, which kills at a rate exceeding that of most cancers. Prevention is therefore of the utmost importance and has to come primarily from a reduction in the incidence and damage done from myocardial infarction. Improved understanding of the pathophysiology of idiopathic dilated cardiomyopathy is also needed. Through understanding of pathophysiologic principles, effective therapy based on interference with known mechanisms that are operative in the disease is now available but can be refined in the future.


A Uniaxial Compression Experiment with CO2-Bearing Coal Using a Visualized and Constant-Volume Gas-Solid Coupling Test System.
Injecting carbon dioxide (CO2) into a deep coal seam is of great significance for reducing the concentration of greenhouse gases in the atmosphere and increasing the recovery of coalbed methane. A visualized and constant-volume gas-solid coupling system is introduced here to investigate the influence of CO2 sorption on the physical and mechanical properties of coal. Being able to keep a constant volume and monitor the sample using a camera, this system offers the potential to improve instrument accuracy and analyze fracture evolution with a fractal geometry method. This paper provides all steps to perform a uniaxial compression experiment with a briquette sample in different CO2 pressures with the gas-solid coupling test system. A briquette, cold-pressed by raw coal and sodium humate cement, is loaded in high-pressure CO2, and its surface is monitored in real-time using a camera. However, the similarity between the briquette and the raw coal still needs improvement, and a flammable gas such as methane (CH4) cannot be injected for the test. The results show that CO2 sorption leads to peak strength and elastic modulus reduction of the briquette, and the fracture evolution of the briquette in a failure state indicates fractal characteristics. The strength, elastic modulus, and fractal dimension are all correlated with CO2 pressure but not with a linear correlation. The visualized and constant-volume gas-solid coupling test system can serve as a platform for experimental research about rock mechanics considering the multifield coupling effect.


Advanced mercury removal from gold leachate solutions prior to gold and silver extraction: a field study from an active gold mine in Peru.
Mercury contamination in the Gold-Cyanide Process (GCP) is a serious health and environmental problem. Following the heap leaching of gold and silver ores with NaCN solutions, portions of the mercury-cyano complexes often adhere to the activated carbon (AC) used to extract the gold. During the electrowinning and retorting steps, mercury can be (and often is) emitted to the air as a vapor. This poses a severe health hazard to plant workers and the local environment. Additional concerns relate to the safety of workers when handling the mercury-laden AC. Currently, mercury treatment from the heap leach solution is nonexistent. This is due to the fact that chelating ligands which can effectively work under the adverse pH conditions (as present in the heap leachate solutions) do not exist. In an effort to economically and effectively treat the leachate solution prior to passing over the AC, a dipotassium salt of 1,3-benzenediamidoethanethiol (BDET2-) has been developed to irreversibly bind and precipitate the mercury. The ligand has proven to be highly effective by selectively reducing mercury levels from average initial concentrations of 34.5 ppm (parts per million) to 0.014 ppm within 10 min and to 0.008 ppm within 15 min. X-ray powder diffraction (XRD), proton nuclear magnetic resonance (1H NMR), Raman, and infrared (IR) spectroscopy demonstrate the formation of a mercury-ligand compound, which remains insoluble over pH ranges of 0.0-14.0. Leachate samples from an active gold mine in Peru have been analyzed using cold vapor atomic fluorescence (CVAF) and inductively coupled plasma optical emission spectroscopy (ICP-OES) for metal concentrations before and after treatment with the BDET2- ligand.


Sex distinctions regarding serum zinc levels in critically ill COVID-19 patients.
Men and women exhibit different presentations in COVID-19. In X chromosome, changes in zinc finger domains cause disorders of sex development. So, we aimed to evaluate sex distinctions regarding serum zinc in severe COVID-19. Data from electronic records of severe COVID-19 patients were correlated with serum zinc. Logistic regression investigated predictors and protectors of hypozincemia in men and women. We assessed 188 medical records (men = 114, women = 74). In men, low zinc was correlated with hypertension (cc = 0.303, p < 0.001), diabetes (cc = 0.198, p = 0.031), hemoglobin (cc = -0.258, p = 0.005), and albumin (cc = -0.219, p = 0.027). Low lymphocyte count (cc = 0.315, p = 0.005), C-reactive protein (cc = -0.248, p = 0.037), and enteral nutrition (cc = 0.269, p = 0.016) were correlated with hypozincemia in women. Age correlated with low zinc in men (c = -0.304, p = 0.001) and women (cc = -0.298, p = 0.010). In men, hypertension (OR = 4.905, p = 0.005) and lymphopenia (OR = -0.999, p = 0.019) were low zinc predictors, while lung injury > 50% was a protective factor (OR = -0.280, p = 0.025). Lymphopenia (OR = -0.999, p = 0.005) and difficult weaning from mechanical ventilation (MV) (OR = 4.359, p = 0.036) were predictors of hypozincemia in women. Difficult weaning from MV (OR = 3.012, p = 0.003) and age (OR = 1.038, p = 0.002) were hypozincemia predictors regardless sex. Hypertension, diabetes, hemoglobin and albumin were correlated with low zinc in men. Lymphopenia, reactive-C protein and enteral nutrition were correlated with low zinc in women. In men, hypertension and low lymphocytes were predictors of hypozincemia. Lymphopenia and difficult weaning from MV were predictors of low zinc in women.


Metabolic profiles of exercise in patients with McArdle disease or mitochondrial myopathy.
McArdle disease and mitochondrial myopathy impair muscle oxidative phosphorylation (OXPHOS) by distinct mechanisms: the former by restricting oxidative substrate availability caused by blocked glycogen breakdown, the latter because of intrinsic respiratory chain defects. We applied metabolic profiling to systematically interrogate these disorders at rest, when muscle symptoms are typically minimal, and with exercise, when symptoms of premature fatigue and potential muscle injury are unmasked. At rest, patients with mitochondrial disease exhibit elevated lactate and reduced uridine; in McArdle disease purine nucleotide metabolites, including xanthine, hypoxanthine, and inosine are elevated. During exercise, glycolytic intermediates, TCA cycle intermediates, and pantothenate expand dramatically in both mitochondrial disease and control subjects. In contrast, in McArdle disease, these metabolites remain unchanged from rest; but urea cycle intermediates are increased, likely attributable to increased ammonia production as a result of exaggerated purine degradation. Our results establish skeletal muscle glycogen as the source of TCA cycle expansion that normally accompanies exercise and imply that impaired TCA cycle flux is a central mechanism of restricted oxidative capacity in this disorder. Finally, we report that resting levels of long-chain triacylglycerols in mitochondrial myopathy correlate with the severity of OXPHOS dysfunction, as indicated by the level of impaired O2 extraction from arterial blood during peak exercise. Our integrated analysis of exercise and metabolism provides unique insights into the biochemical basis of these muscle oxidative defects, with potential implications for their clinical management.


Stem cell transplantation for multiple sclerosis: what is the evidence?
Experimental and clinical observations have indicated that high-dose immunosuppression followed by autologous stem cell transplantation (ASCT) can induce remissions in severe, refractory, autoimmune diseases including multiple sclerosis (MS), a T cell-mediated autoimmune disorder against CNS myelin components, causing severe chronic disability. Control of the disease is unsatisfactory in most of the patients, especially those with rapidly evolving relapsing-remitting course and those with chronic progressive disease. The rationale for treating autoimmune diseases with ASCT is based on the immunosuppressive and immunomodulating effects of ASCT which may shift the immunological balance towards disease quiescence, a hypothesis supported by the results of ASCT in animal models of MS and by clinical observations in MS patients transplanted for concurrent malignancies. A number of phase I-II studies of ASCT in patients with active MS, conducted worldwide since 1995, and a comprehensive analysis of 85 patients, recently reported by the European Group for Blood and Marrow Transplantation (EBMT), have shown the feasibility of the method, a prominent anti-inflammatory effect on magnetic resonance imaging (MRI) disease, and a possible clinical benefit for active and refractory cases. The impact on MRI disease parameters appears superior with ASCT than with conventional therapies but the clinical results, in terms of stabilization of disease and prevention of disability, need to be validated in prospective, controlled trials. The procedure is also associated with a transplant-related mortality risk, of about 5% in high-risk cases, i.e., in older patients, those with high disability scores, those receiving strong myeloablative conditioning regimens and those undergoing intensive in vivo or ex vivo T cell-depletion. Therefore, it could be recommended for the treatment of a chronic, non-lethal, disease like MS only if it proved superior to standard therapies. A randomized trial is now launched by the EBMT to compare ASCT to mitoxantrone, currently regarded as one of the best available treatments, in properly selected patients having high chance of response at minimal mortality risk.


Ho-Ion-Polymer/Graphene Heterojunctions Toward Room-Temperature Ferromagnets.
Organic ferromagnetic materials offer great promise for spintronic devices, carbon-based chips, and quantum communications, but remain as a challenging issue due to their low saturation magnetization and/or unsustainable ferromagnetic properties. To date, magnetic ion polymers have displayed paramagnetism without exception at room-temperature. In this study, it is reported for the first time that, owing to the structural restriction and charge exchange of Ho ion by polymer/graphene π-π stacking heterojunctions, holmium ion polymer composites exhibited typical hysteresis lines of ferromagnetic materials at room temperature. The room-temperature ferromagnetic ion polymer composite presented the highest saturation magnetization value of 3.36 emu g-1 and unprecedented sustainable ferromagnetism, compared to reported room-temperature organic ferromagnetic materials. Accordingly, prepared ferromagnetic composites also achieved impressive wave absorption properties, with a maximum reflection loss of as much as -57.32 dB and a broad absorption bandwidth of 5.05 GHz. These findings may promote the development of room-temperature organic ferromagnetic materials.


Mechanism of pain in osteoid osteomas: an immunohistochemical study.
Osteoid osteoma is a benign bone tumour characterized by pain which is relieved by non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin. To clarify the mechanism of the pain, five osteoid osteomas were studied immunohistochemically using polyclonal antibodies against prostaglandin E2 (PGE2), S-100 protein and protein gene product 9.5 (PGP 9.5). In all five cases, the pain had been relieved by NSAIDs. Nerve fibres positive for S-100 protein and PGP 9.5 were observed in the fibrous zone, especially close to the blood vessels, around the nidus in all the lesions and also within the nidus in three lesions. PGE2 immunoreactivity was variably positive in the nidus of three lesions. In one case a large number of actively proliferating osteoblasts reacted with this antibody. The other cases showed unevenly distributed PGE2 positivity which tended to be prominent in the plump osteoblasts. As control material we examined fibrous dysplasia (3 cases), osteosarcomas (3) and giant-cell tumours of bone (3). The plump osteoblastic tumour cells of three osteosarcomas and the bone-forming cells in two cases of fibrous dysplasia gave a positive reaction for PGE2. No S-100 or PGP 9.5 immunoreactive nerve fibres were seen in these lesions. It is concluded that the presence of nerve fibres alone might play a more important role in mediation of pain in osteoid osteomas than some effects of osteoblast-produced PGE2 on the nerves and proliferated blood vessels.


Use of health services according to income before and after elimination of copayment in Germany and restriction of universal health coverage in Spain.
In Germany copayment for medical consultation was eliminated in 2013, and in Spain universal health coverage was partly restricted in 2012. This study shows the relationship between income and the use of health services before and after these measures in each country. Data were taken from the 2009 and 2014 Socio-Economic Panel conducted in Germany, and from the 2009 and 2014 European Health Surveys in Spain. The health services investigated were physician consultations and hospital admissions, and the measure of socioeconomic position used was household income. The magnitude of the relationship between socioeconomic position and the use of each health service in people from 16 to 74 years old was estimated by calculating the percentage ratio using binary regression. In Germany, after adjusting for age, sex, and need for care, in the model comparing the two lower income categories to the two higher categories, the percentage ratio for physician consultation was 0.97 (95% CI 0.96-0.99) in 2009 and 0.98 (95% CI 0.97-0.99) in 2014, and the percentage ratio for hospitalization was 1.01 (95% CI 0.93-1.10) in 2009 and 1.16 (95% CI 1.08-1.25) in 2014. In Spain, after adjusting for age, sex, and self-rated health, the percentage ratio for physician consultation was 0.99 (95% CI 0.94-1.05) in 2009 and 1.08 (95% CI 1.03-1.14) in 2014, and the percentage ratio for hospitalization was 1.04 (95% CI 0.92-1.18) in 2009 and 0.99 (95% CI 0.87-1.14) in 2014. The results suggest that elimination of the copayment in Germany did not change the frequency of physician consultations, whereas after the restriction of universal health coverage in Spain, subjects with lower incomes had a higher frequency of physician consultations.


Temporal Binding Window of the Sound-Induced Flash Illusion in Amblyopia.
Amblyopia is a neurodevelopmental visual disorder caused by abnormal visual experience in childhood. In addition to known visual deficits, there is evidence for changes in audiovisual integration in amblyopia using explicit tasks. We examined audiovisual integration in amblyopia using an implicit task that is more relevant in a real-world context. A total of 11 participants with amblyopia and 16 controls were tested binocularly and monocularly on the sound-induced flash illusion, in which flashes and beeps are presented concurrently and the perceived number of flashes is influenced by the number of beeps. The task used 1 to 2 rapid peripheral flashes presented with 0 to 2 beeps, at 5 stimulus onset asynchronies, that is, beep (-200 milliseconds, -100 milliseconds) or flash leading (100 milliseconds, 200 milliseconds) or simultaneous (0 milliseconds). Participants reported the number of perceived flashes. Susceptibility was indicated by a "2 flashes" response to "fission" (1 flash, 2 beeps) or "1 flash" to "fusion" (2 flashes, 1 beep). For fission with the beep leading during binocular viewing, controls showed an expected decrease in illusion strength as stimulus onset asynchronies increased, whereas the illusion strength remained constant in participants with amblyopia, indicating a wider temporal binding window in amblyopia (P = 0.007). For fusion, participants with amblyopia showed reduced illusion strength during amblyopic eye viewing (P = 0.044) with the flash leading. Amblyopia is associated with the widening of the temporal binding window, specifically for fission when viewing binocularly with the beep leading. This suggests a developmental adaptation to delayed amblyopic eye visual processing to optimize audiovisual integration.


Sphingolipid biosynthetic pathway genes FEN1 and SUR4 modulate amphotericin B resistance.
Deletants of the sphingolipid biosynthetic pathway genes FEN1 and SUR4 of Saccharomyces cerevisiae, as well as deletants of their orthologs in Candida albicans, were found to be 2- to 5-fold-more sensitive to amphotericin B (AmB) than parent strains. The inhibition of sphingolipid biosynthesis in parent strains by myriocin sensitized them to AmB, which can be reversed by providing phytosphingosine, an intermediate in the sphingolipid pathway. These results indicate that sphingolipids modulate AmB resistance, with implications for mechanisms underlying AmB action and resistance.


Ulinastatin protects against sepsis‑induced myocardial injury by inhibiting NLRP3 inflammasome activation.
Myocardial injury is the primary manifestation of multiple organ dysfunction during sepsis, however, the mechanisms underlying sepsis‑induced myocardial injury remain unclear. Similarly, no effective therapeutics have yet been developed for myocardial injury. In the present study, the role of the NOD‑like receptor 3 (NLRP3) inflammasome on cardiac function were characterized and the effects of different ulinastatin (UTI) doses in protecting a septic rat model from myocardial injury were elucidated. To evaluate UTI efficacy on cardiac function, its effects on anti‑inflammatory mediators were analyzed and its cardioprotective effects were investigated. It was demonstrated that circulatory levels of tumor necrosis factor‑α and interleukin‑1β were elevated during sepsis. It was also observed that NLRP3 and caspase‑1 expression enhanced post‑cecal ligation and puncture (CLP), and that high UTI levels protected against myocardial injury induced by sepsis. To the best of our knowledge, this is the first study to demonstrate that the mechanisms underpinning UTI‑mediated myocardial protection were due to the downregulation of the NLRP3/caspase‑1/IL‑1β signaling pathway. Based on these findings, it is proposed that UTI exerts beneficial effects during sepsis‑induced myocardial injury.


A retrospective, hospital-based study to determine the incidence of rotavirus hospitalizations among children less than 5 years of age over a 10-year period (2001-2011) in Akita prefecture, Japan.
Rotavirus is the most common cause of severe gastroenteritis in children worldwide. This retrospective, cross-sectional study was undertaken in a sentinel hospital that provides the only pediatric beds for the local population with an average of 4,400 children aged <5 years and determined the incidence of rotavirus hospitalizations. Medical charts that recorded acute gastroenteritis cases occurring in children aged <5 years living in the cites of Yuri-Honjo or Nikaho, Akita, Japan between 2001 and 2011 were retrieved and examined to enumerate rotavirus antigen-positive hospitalizations. Of the 1,596 acute gastroenteritis cases retrieved, antigen detection was performed in 834 cases, and 387 were positive; hence, the crude annual incidence rate of rotavirus hospitalizations was 8.8 per 1,000 person-years. The adjusted annual incidence rate of rotavirus hospitalizations was 13.7 per 1,000 person-years when untested samples collected during the peak season were extrapolated to the same rotavirus detection proportion as the tested samples (58.9%). We confirmed a high incidence of rotavirus hospitalizations in Akita Prefecture and revealed a considerable degree of annual fluctuation in the rotavirus hospitalization rates, which exceeded the degree of stochastic fluctuation. Thus, caution must be exercised when interpreting the impact of a rotavirus vaccine on the reduction of the number of rotavirus hospitalizations.


Identification of a novel first exon of prolactin receptor gene expressed in the rat brain.
A novel first exon, E1(4), whose sequence was distinct from those of the three known first exons, E1(1), E1(2), and E1(3), of the rat PRL receptor (PRL-R) gene was identified by cDNA cloning for the 5'-end region of PRL-R mRNA expressed in the rat brain. Sequence analysis revealed the presence of two different length E1(4) cDNAs. The longer cDNA contained the 243-bp E1(4) sequence, and the shorter cDNA lacked the 139-bp sequence at the 5'-end of the longer one. Neither E1(4) cDNA has a second exon sequence, indicating that the E1(4) first exon is extensively spliced to the third exon. E1(4)-containing PRL-R mRNAs were detected only in the brain by RT-PCR and ribonuclease protection assay. The longer E1(4) mRNA was expressed as the major PRL-R mRNA species in the brain and was greatly increased in pregnant (d 18) and lactating (d 5) rats. A genomic clone containing the E1(4) first exon together with its 5'- and 3'-flanking regions was isolated from a rat kidney genomic library. Ribonuclease protection assay revealed that the position corresponding to the 5'-end of the shorter E1(4) cDNA is the major transcription start point for the E1(4) exon. The 5'-flanking region of E1(4) contained a TATA box-like element 23 bp upstream of the major transcription start point. Other putative transcription factor-binding sites, such as CCAAT, Sp1, and glucocorticoid-responsive elements, were observed at further upstream regions. These results suggest that PRL-R gene expression in rat brain is controlled by the promoter for the E1(4) first exon.


'A child with Malpuech-Michels-Mingarelli-Carnevale syndrome and ADHD and major depressive disorder'.
Malpuech-Michels-Mingarelli-Carnevale (3MC) syndrome, is a rare genetic condition resulting from the combination of four autosomal recessive syndromes which were classified as separate syndromes earlier. 3MC syndrome may be accompanied by a range of other conditions including cleft lips and palate, blepharophimosis, blepharoptosis, downward-sloping palpebral fissures, hypertelorism, facial dysmorphism such as high arched eyebrows, growth retardation, hearing impairment, genital anomalies, elongated coccyx, caudal appendage, radioulnar synostosis and skeletal conditions such as craniosynostosis. The prominent causes of 3MC syndrome include homozygous mutations in the MASP1, COLEC10, or COLEC11 genes. Few cases with 3MC syndrome have been reported in the literature. Here we present a case of 11-year-old girl with 3 MC syndrome in comorbidity with attention deficit hyperactivity disorder, oppositional defiant disorder, and major depressive disorder.


Effect of 2-deoxyglucose on Schizosaccharomyces pombe.
Megnet, Roland (Institut für Allgemeine Mikrobiologie der Universität, Bern, Switzerland). Effect of 2-deoxyglucose on Schizosaccharomyces pombe. J. Bacteriol. 90:1032-1035. 1965.-Cultivation of Schizosaccaromyces pombe in a medium containing 2-deoxyglucose (100 mug/ml) results in the death of the cells after an initial period of apparently normal growth. At higher deoxyglucose concentration (400 mug/ml), the cells die immediately after inoculation. Only growing cells are killed, and microscopic inspection of the cultures reveals cell-wall fragments of lysed cells. A mutant resistant to 2-deoxyglucose, which cannot use glucose as a carbon source, was found to be partially deficient in hexokinase. The data constitute evidence for the inhibition of some reaction(s) in the synthesis of cell-wall polysaccharides by metabolites of 2-deoxyglucose in this organism.


Boost glycemic control in type 2 diabetics with stress management training.
Why? Because studies confirm that stress can have an adverse effect on glycemic control. Individualized therapy has been proven effective in such cases, but now for the first time one study shows that a more practical, group approach to stress management can produce impressive results as well.


Usefulness of the convexity apparent hyperperfusion sign in 123I-iodoamphetamine brain perfusion SPECT for the diagnosis of idiopathic normal pressure hydrocephalus.
The gold standard for the diagnosis of idiopathic normal pressure hydrocephalus (iNPH) is the CSF removal test. For elderly patients, however, a less invasive diagnostic method is required. On MRI, high-convexity tightness was reported to be an important finding for the diagnosis of iNPH. On SPECT, patients with iNPH often show hyperperfusion of the high-convexity area. The authors tested 2 hypotheses regarding the SPECT finding: 1) it is relative hyperperfusion reflecting the increased gray matter density of the convexity, and 2) it is useful for the diagnosis of iNPH. The authors termed the SPECT finding the convexity apparent hyperperfusion (CAPPAH) sign. Two clinical studies were conducted. In study 1, SPECT was performed for 20 patients suspected of having iNPH, and regional cerebral blood flow (rCBF) of the high-convexity area was examined using quantitative analysis. Clinical differences between patients with the CAPPAH sign (CAP) and those without it (NCAP) were also compared. In study 2, the CAPPAH sign was retrospectively assessed in 30 patients with iNPH and 19 healthy controls using SPECT images and 3D stereotactic surface projection. In study 1, rCBF of the high-convexity area of the CAP group was calculated as 35.2–43.7 ml/min/100 g, which is not higher than normal values of rCBF determined by SPECT. The NCAP group showed lower cognitive function and weaker responses to the removal of CSF than the CAP group. In study 2, the CAPPAH sign was positive only in patients with iNPH (24/30) and not in controls (sensitivity 80%, specificity 100%). The coincidence rate between tight high convexity on MRI and the CAPPAH sign was very high (28/30). Patients with iNPH showed hyperperfusion of the high-convexity area on SPECT; however, the presence of the CAPPAH sign did not indicate real hyperperfusion of rCBF in the high-convexity area. The authors speculated that patients with iNPH without the CAPPAH sign, despite showing tight high convexity on MRI, might have comorbidities such as Alzheimer's disease.