Antioxidant and cyclooxygenase activities of fatty acids found in food.
Several commercially available C-8 to C-24 saturated and unsaturated fatty acids (1-29) were assayed for cyclooxygenase-I (COX-I) and cyclooxygenase-II (COX-II) inhibitory and antioxidant activities. Among the saturated fatty acids tested at 60 microg mL(-1), there was an increase in antioxidant activity with increasing chain length from octanoic acid to myristic acid (C-8-C-14) and a decrease thereafter. All unsaturated fatty acids tested at 60 microg mL(-1) showed good antioxidant activity except for undecylenic acid (12), cis-5-dodecenoic acid (13), and nervonic acid (29). The highest inhibitory activities among the saturated fatty acids tested on cyclooxygenase enzymes COX-I and COX-II were observed for decanoic acid to lauric acid (3-5) at 100 microg mL(-1). Similarly, among the unsaturated fatty acids tested, the highest activities were observed for cis-8,11,14-eicosatrienoic acid (25) and cis-13,16-docosadienoic acid (27) at 100 microg mL(-1).


[Face trauma in young people: medical and social aspects of the problem].
Problem of maxillofacial traumatism in juvenile age group has not only medical but also social aspect. The social aspect of the problem is in traumatism steady growth in this most vulnerable in the social attitude population group that lead not only to considerable disability but to significant society expenditures on patients rehabilitation. Medical aspect of the problem is in the special character of young organism biological reaction upon trauma that should be taken into consideration when organizing medical care to the victims.


Treatment of aggression in children and adolescents with autism and conduct disorder.
The optimal clinical management of aggression in children and adolescents involves both behavioral and pharmacologic intervention strategies. This article reviews medication treatments for youngsters with autistic disorder and conduct disorder, conditions for which the pharmacologic management of aggression is often necessary. Efficacy results and associated adverse effects from selected clinical trials of most classes of psychotropic medications are discussed. While preliminary progress has been made in the development of medication treatments for these serious disorders of youth, additional controlled research and longitudinal studies are needed to better understand the efficacy and tolerability of currently available compounds within each diagnostic group.


Non-glycosylated tandem repeats of MUC1 facilitate attachment of breast tumor cells to normal human lung tissue and immobilized extracellular matrix proteins (ECM) in vitro: potential role in metastasis.
MUC1 is a transmembrane glycoprotein abundantly expressed on the apical surface of human ductal epithelial cells and over entire cell surface of tumors originating from those cells. It is 300 to 500 nm long and has a rigid, rod-like structure protruding from the cell surface. MUC1 expressed by normal cells has heavily O-glycosylated tandem repeat domain while MUC1 on malignant cells is aberrantly O-glycosylated. Substantially reduced (aberrant) glycosylation of the tandem repeat region of tumor MUC1 results in uncovering of the polypeptide core. This new structural feature may play an important role in the attachment of metastasizing tumor cells to tissues at distant sites. We show that MDA-MB-231 cells attaching to the immobilized extracellular matrix proteins (ECM) are higher MUC1 expressers than those non-attaching and that the attachment is inhibited by the addition of non-glycosylated, MUC1 peptide. This 100 a.a. peptide composed of 5 tandem repeats from the tandem repeat domain mimics the forms of MUC1 found in ascites fluid of cancer patients. We also show that this synthetic form of MUC1 inhibited attachment of breast tumor cells to sections of normal human lung tissue and immobilized ECM. We did not find correlation between the expression of Tn (GalNAc-Ser/Thr) epitope and the ability of tumor cells to adhere to the immobilized ECM. These results indicate that the non-glycosylated form of MUC1 plays a role in the initial attachment of carcinoma cells to tissues at distant sites, which may facilitate establishment of metastatic foci.


Photo-redox reactivity of titanium-oxo clusters: mechanistic insight into a two-electron intramolecular process, and structural characterisation of mixed-valent Ti(iii)/Ti(iv) products.
Small titanium-oxo-alkoxide clusters, [TiO(OR)(O2PR'2)]4, synthesised by the stoichiometric reaction of Ti(OiPr)4, phosphinic acid and water, undergo a photo-redox transformation under long-wave UV light. The photo-reaction generates blue coloured, mixed-valence Ti(iii)/Ti(iv)-oxo clusters alongside acetone and isopropanol by-products. This reactivity indicates the ability for photoactivated charge separation to occur in even the smallest of Ti-oxo clusters. EPR and NMR spectroscopic studies support a photo-redox mechanism that occurs via an intramolecular, two-electron pathway, directly relating to current doubling effects observed at TiO2 photoanodes in the presence of alcohols. The rate of photo-reaction is solvent dependent, with donor solvents supporting the formation of low coordinate Ti(iii) sites. The nature of the electronic transition is identified by DFT and TDDFT calculations as an oxygen to titanium charge transfer and it is possible to finetune the UV absorption onset observed by changing the phosphinate ligand. A two-electron photo-reduced cluster, [Ti4O4(O2PPh2)6], forms spontaneously from the photo-reaction and its structure is identified by X-ray crystallography with supporting DFT calculations. These indicate that [Ti4O4(O2PPh2)6] is high-spin and contains two ferromagnetically coupled electrons delocalised over the Ti4 core. [Ti4O4(O2PPh2)6] undergoes rapid oxidation in air in the solid-state and performs a remarkable single-crystal to single-crystal transformation, to form a stable cluster-superoxide salt.


Extended phage locus typing of Salmonella enterica serovar Typhimurium, using multiplex PCR-based reverse line blot hybridization.
Salmonella enterica serovar Typhimurium (S. Typhimurium) is the commonest pathogen causing food-borne disease among humans and animals in Australia. A multiplex PCR-based reverse line blot (mPCR/RLB) system was developed to rapidly identify S. Typhimurium phage types and strains within them. The system comprised 32 biotin-labelled primer sets and 38 amino-labelled probes, based on sequences that were either phage-type-related or derived from temperate phages ST64B, P22, Gifsy-1 or Gifsy-2. The system was developed and evaluated using 168 S. Typhimurium isolates, representing 46 phage types. RLB patterns, based on a combination of positive hybridization and grading of signal intensities, validated by sequencing, differentiated S. Typhimurium isolates into 102 types. Some clusters contained isolates belonging to a single phage type while others contained isolates belonging to more than one. Most phage types exhibited at least two RLB profiles. The feasibility of this system was evaluated during investigations of three outbreaks, due to two different phage types. Within each outbreak, isolates showed identical RLB patterns, whereas sporadic isolates of corresponding phage types showed various patterns. The mPCR/RLB system was compared with multilocus variable-number tandem-repeat analysis (MLVA). The two methods demonstrated similar discriminatory abilities. Based on these preliminary results, the mPCR/RLB system is a promising tool for molecular identification of most common S. Typhimurium phage types. It could be used as an alternative to, or in conjunction with, MLVA for rapid strain typing during outbreaks.


Towards a dynamical network view of brain ischemia and reperfusion. Part IV: additional considerations.
The general failure of neuroprotectants in clinical trials of ischemic stroke points to the possibility of a fundamental blind spot in the current conception of ischemic brain injury, the "ischemic cascade". This is the fourth in a series of four papers whose purpose is to work towards a revision of the concept of brain ischemia by applying network concepts to develop a bistable model of brain ischemia. Here we consider additional issues to round out and close out this initial presentation of the bistable network view of brain ischemia. Initial considerations of the network architecture underlying the post-ischemic state space are discussed. Network and differential equation models of brain ischemia are compared. We offer a first look at applying the bistable model to focal cerebral ischemia. The limitations of the present formulation of the bistable model are discussed. This work concludes with a series of questions by which to direct future efforts.


Preparation and properties of immune-stimulating complexes containing hepatitis B virus surface antigen.
Immune-stimulating complexes (iscoms) have been prepared containing the major S gene products (HBsAg) of the hepatitis B virus genome. Immunization of BALB/c mice with a single dose of hepatitis B iscoms in saline resulted in a high titre antibody response to HBsAg. In contrast, the original HBsAg preparation required an adjuvant to produce equivalent amounts of antibody. Analysis of sera from mice immunized with hepatitis B iscoms revealed antibodies directed against the major a determinants of HBsAg. High secondary antibody responses were observed in immunized animals previously inoculated with a sub-immunogenic dose of HBsAg indicating that hepatitis B iscoms may represent a suitable immunogen for use in individuals in whom a course of immunization with currently licensed hepatitis B vaccines has failed to produce a significant anti-HBs response.


Prognostic significance of the number of postoperative intraperitoneal chemotherapy cycles for patients with advanced epithelial ovarian cancer.
Phase 3 trials have demonstrated a survival advantage for patients with optimally debulked epithelial ovarian cancer who received intravenous (IV) and intraperitoneal (IP) chemotherapy compared with IV therapy alone. This was despite a significant proportion of patients in the IV/IP arms not completing all 6 planned cycles. Our objective was to evaluate the prognostic significance of the number of IV/IP cycles administered. Data were analyzed for all patients with stage III to IV epithelial ovarian cancer who underwent optimal primary cytoreduction followed by 1 or more cycles of IV/IP chemotherapy from January 2005 to July 2011 at our institution. A landmark analysis was performed to associate progression-free survival (PFS) and overall survival (OS) with the number of IV/IP cycles given. We identified 201 patients; 26 (13%) received 1 to 2 cycles of IV/IP chemotherapy, 41 (20%) received 3 to 4 cycles, and 134 (67%) received 5 to 6 cycles. The 5-year PFS for patients who received 1 to 2, 3 to 4, and 5 to 6 cycles was 18%, 29%, and 17%, respectively. The 5-year OS for patients who received 1 to 2, 3 to 4, and 5 to 6 cycles was 44%, 54%, and 57%, respectively. There was no significant difference in PFS (P = 0.31) or OS (P = 0.14) between the 3 groups. The most common reason for discontinuing IV/IP therapy was treatment-related toxicity (77%). Postoperative complications were the most common reason for not initiating IV/IP therapy (42%) in patients who subsequently transitioned to it. We did not detect a significant survival difference between patients who received 1 to 2, 3 to 4, or 5 to 6 IV/IP chemotherapy cycles. Women may still derive a survival benefit if they receive fewer than 6 IV/IP cycles.


[Study of toxicity and antineoplastic activity of 14-substituted derivatives of rubomycin and carminomycin].
Toxicity and antitumor activity of five derivatives of rubomycin and carminomycin were studied in animals. The derivatives were prepared by modification of the methyl C-14 group. These were the following: 14-chlorrubomycin, 14-chlorcarminomycin, 14-salicyloyloxyrubomycin, 14-salicyloyloxycarminomycin and 14-quinaldinoyloxyrubomycin. The chemotherapeutic study revealed that, in their activity, all the compounds were inferior to the starting antibiotics. Unlike the other derivatives, 14-chlorcarminomycin induced a significant inhibition of leukemia P-388 development (the average lifespan of the mice amounted to 165 per cent as compared to the control). However, in the magnitude of its effect, the derivative was inferior to carminomycin.


[Guanylate cyclase activity of the rabbit myocardium in normal conditions and in allergic states].
Activity of guanylate cyclase, regulation of the enzyme by means of hormones and other biologically active substances as well as level of cGMP were studied in rabbit myocardium of control animals and in animals with allergic impairment of the tissue. The enzyme was shown to exhibit no sensitivity to acetylcholine and histamine, which appears to occur as a result of the hormone-unspecific desensitization of the corresponding receptors in allergic impairment. Decreased basal activity of guanylate cyclase was accompanied by an increase in content of cGMP in myocardial tissue. This phenomenon was apparently also due to a decrease in phosphodiesterase activity, which is observed under the conditions of this pathology. At the same time, the decrease in the basal activity of guanylate cyclase was caused by impairment in active restoration of the enzyme in the allergic tissue. A decrease in the enzyme activation by means of azide and nitroprusside appears to be due to loss of hem, prosthetic group of the cyclase. Physiological functions of guanylate cyclase are discussed.


Ligand- and species-dependent activation of PPARalpha.
Peroxisome proliferator-activated receptor alpha (PPARalpha) is mainly expressed in liver and involved in lipid metabolism. Oxidation of certain fatty acids in peroxisomes is under PPARalpha control. A wide variety of lipid molecules activate PPARalpha as well as the fibric acid derivative clofibrate. In the present study, we evaluated the differential activation of PPARalpha with several agonist ligands through its expression and DNA binding in both rat (McA-RH7777) and human (HepG2) hepatoma cell lines. In McA-RH7777 cells, clofibrate alone mediated a higher induction of PPARalpha expression than linoleic acid. In contrast, linoleic acid was the most effective ligand in HepG2 cells and treatment with clofibrate plus linoleic acid did not further increase PPARalpha expression. PPRE-binding activity of PPARalpha in ligand-treated cells was also increased in a parallel manner. We suggest that ligand-induced PPARalpha activation might give rise to differential species-dependent responses.


Reovirus cell entry requires functional microtubules.
Mammalian reovirus binds to cell-surface glycans and junctional adhesion molecule A and enters cells by receptor-mediated endocytosis in a process dependent on β1 integrin. Within the endocytic compartment, reovirus undergoes stepwise disassembly, allowing release of the transcriptionally active viral core into the cytoplasm. To identify cellular mediators of reovirus infectivity, we screened a library of small-molecule inhibitors for the capacity to block virus-induced cytotoxicity. In this screen, reovirus-induced cell killing was dampened by several compounds known to impair microtubule dynamics. Microtubule inhibitors were assessed for blockade of various stages of the reovirus life cycle. While these drugs did not alter reovirus cell attachment or internalization, microtubule inhibitors diminished viral disassembly kinetics with a concomitant decrease in infectivity. Reovirus virions colocalize with microtubules and microtubule motor dynein 1 during cell entry, and depolymerization of microtubules results in intracellular aggregation of viral particles. These data indicate that functional microtubules are required for proper sorting of reovirus virions following internalization and point to a new drug target for pathogens that use the endocytic pathway to invade host cells. Screening libraries of well-characterized drugs for antiviral activity enables the rapid characterization of host processes required for viral infectivity and provides new therapeutic applications for established pharmaceuticals. Our finding that microtubule-inhibiting drugs impair reovirus infection identifies a new cell-based antiviral target.


Role of histopathology as an aid to prognosis in rhino-orbito-cerebral zygomycosis.
Rhino-orbito-cerebral zygomycosis is a rapidly progressive opportunistic fungal infection characterized by a set of clinical and radiological findings that help in prognostication. The present study is aimed to evaluate its histopathologic features as an aid to prognosis in order to guide the physician at the stage of tissue diagnosis to optimize surgery, chemotherapy and immunosuppression. The study comprises of a microscopic analysis of specific histopathologic variables on 33 cases of zygomycosis that were diagnosed and treated in a seven-year period. Fungal load in the tissue (graded as mild, moderate and marked), mean diameter of fungus, degree of neutrophilic and granulomatous response, tissue invasion and necrosis were graded and assessed for their prognostic significance. Seasonal variation, signs and symptoms, extent of involvement and laboratory parameters were also analyzed to examine the trend of occurrence of the disease and to associate these with patient's outcome, which was categorized as either survived or expired. The follow-up ranged from 1 month to 7 years. Of all the histological variables, angioinvasion was directly related to the mortality rate. Diameter of the fungal hyphae and its intraorbital or intracranial invasion also proved to be significant indicators of poor prognosis. (P = 0.04 and 0.0037, respectively) though angioinvasion was directly related to the mortality rate. Thus, histopathology could assist the clinician in assessing patient's progress and thus optimize the treatment in such patients.


Pathogenic Roles of the Bacteria carried by Bursaphelenchus mucronatus.
Fifty strains of bacteria were isolated from six isolates of the nematode Bursaphelenchus mucronatus (Bm) from China and Russia and identified using the BioMerieux Vitek 32 system. In bioassay, 3 bacterial strains showed the high levels of phytotoxin production while 19, 16, and 12 strains showed moderately, low and no phytotoxin production, respectively. Inoculation of 2-month-old Pinus thunbergii seedling with each of the six Bm isolates showed that the mean number of days from inoculation to death of 80% of the seedlings was significantly related to the ratio of the total number of bacterial strains for a nematode isolate to the number of pathogenic bacterial strains of the nematode isolate. The results of inoculation of 3-year-old P. thunbergii seedlings showed that inoculation with either axenic Bm (ABm) or axenic B. xylophilus (ABx) and the pathogenic bacterial strain together were essential for inducing pine wilt. These findings demonstrate that wilt symptoms caused by Bm conform to our earlier hypothesis (Zhao et al., 2003) that pine wilt disease, induced by certain Bx or Bm isolates, is caused by a complex of both the nematodes and their associated pathogenic bacteria. The results also account for the variation in pathogenicity of Bm populations from different parts of the world.


A cohort study of MFN2 mutations and phenotypic spectrums in Charcot-Marie-Tooth disease 2A patients.
Charcot-Marie-Tooth disease 2A (CMT2A) is the most common axonal form of peripheral neuropathy caused by a defect in the mitofusin 2 (MFN2) gene, which encodes an outer mitochondrial membrane GTPase. MFN2 mutations result in a large range of phenotypes. This study analyzed the prevalence of MFN2 mutation in Korean families with their assorted phenotypes (607 CMT families and 160 CMT2 families). Direct sequencing of the MFN2 coding exons or whole-exome sequencing has been applied to identify causative mutations. A total of 21 mutations were found in 36 CMT2 families. Comparative genotype-phenotype correlations impacting severity, onset age, and specific symptoms were assessed. Most mutations were seen in the GTPase domain (∼86%). A deletion mutation found in the transmembrane helices is reported for the first time, as well as five novel mutations at other domains. MFN2 mutations made up 5.9% of total CMT families, whereas 22.9% in CMT2 families, of which 27.8% occurred de novo. Interestingly, patient phenotypes ranged from mild to severe even for the same mutation, suggesting other factors influenced phenotype and penetrance. This CMT2A cohort study will be useful for molecular diagnosis and treatment of axonal neuropathy.


Gadobenate dimeglumine and cerebral glucose metabolism. Continuous monitoring of striatal lactate levels in freely moving rats.
Brain contrast-enhanced MR imaging reflects the leakage of contrast material into the brain tissue due to blood-brain barrier (BBB) disruption. The contact between brain tissue and contrast material requires a high level of neurotolerability of the contrast agent (CA). In the present study, we investigated the neurotolerability of the paramagnetic CA gadobenate dimeglumine, locally applied into the corpus striatum of freely moving rats, by evaluating its potential effects on cerebral glucose metabolism based on lactate levels. Lactate levels were monitored using a microdialysis technique coupled with an enzyme reaction. A microdialysis probe for extracellular fluid sampling, together with a stainless steel cannula for CA administration, were inserted into the right corpus striatum of rats. Lactate levels were monitored for 2 h after gadobenate dimeglumine administration at 120 nmol/rat, at fixed volume of 1.2 microl. The same volume of artificial cerebrospinal fluid (aCSF) was administered to control rats. Gadobenate dimeglumine did not induce any significant changes in the lactate striatal levels over the 30-min period after administration. Small, but significant, reductions in lactate concentration were found from the 45-min control point after gadobenate dimeglumine administration. Lactate response showed the same pattern in rats given aCSF. Gadobenate dimeglumine, intracerebrally administered, did not affect cerebral glucose metabolism in rats as it showed the same behaviour as aCSF on cerebral glucose utilization. The gradual attenuation in the endogenous lactate release observed 45 min after test compound administration is possibly due to a slight reduction in the probe recovery. The present findings confirm the neurotolerability of gadobenate dimeglumine previously shown in behavioural and electrophysiological studies.


Transcriptional interference and gene orientation in yeast: noncoding RNA connections.
The compressed nature of genes in yeast requires that transcription units be well defined to prevent read-though transcription from one gene into an adjacent gene. Failure to terminate transcription may result in transcriptional interference of downstream-positioned genes. Transcriptional interference has been studied in several budding yeast gene systems and, interestingly, may be used as a general mechanism of gene regulation. The realization that extensive numbers of noncoding RNAs are transcribed between and across protein-coding genes greatly increases the complexity and subtlety of gene regulation through transcriptional interference. This may be achieved by somehow directly blocking RNA polymerase access to promoters or by the formation of repressive chromatin structures. Furthermore, in fission yeast, read-through transcription from convergent genes may generate double-strand RNAs. These will elicit RNA interference, resulting in heterochromatin formation and consequent gene silencing. Although much remains to be learned from yeast, it is apparent that higher eukaryotes also use related transcriptional interference and gene-silencing strategies. Even though protein-coding genes in mammals are widely separated along chromosomes, extensive numbers of noncoding RNAs are also synthesized. These may well connect distant genes and thus promote transcriptional interference and gene silencing, as is now well established for yeast.


Directed chromosomal integration and expression of the reporter gene gusA3 in Lactobacillus acidophilus NCFM.
Lactobacillus acidophilus NCFM is a probiotic microbe that survives passage through the human gastrointestinal tract and interacts with the host epithelium and mucosal immune cells. The potential for L. acidophilus to express antigens at mucosal surfaces has been investigated with various antigens and plasmid expression vectors. Plasmid instability and antibiotic selection complicate the possibility of testing these constructs in human clinical trials. Integrating antigen encoding genes into the chromosome for expression is expected to eliminate selection requirements and provide genetic stability. In this work, a reporter gene encoding a β-glucuronidase (GusA3) was integrated into four intergenic chromosomal locations. The integrants were tested for genetic stability and GusA3 activity. Two locations were selected for insertion downstream of constitutively highly expressed genes, one downstream of slpA (LBA0169), encoding a highly expressed surface-layer protein, and one downstream of phosphopyruvate hydratase (LBA0889), a highly expressed gene with homologs in other lactic acid bacteria. An inducible location was selected downstream of lacZ (LBA1462), encoding a β-galactosidase. A fourth location was selected in a low-expression region. The expression of gusA3 was evaluated from each location by measuring GusA3 activity on 4-methyl-umbelliferyl-β-d-glucuronide (MUG). GusA3 activity from both highly expressed loci was more than three logs higher than the gusA3-negative parent, L. acidophilus NCK1909. GusA3 activity from the lacZ locus was one log higher in cells grown in lactose than in glucose. The differences in expression levels between integration locations highlights the importance of rational targeting with gene cassettes intended for chromosomal expression.


Alternative Splice in Alternative Lice.
Genomic and transcriptomics analyses have revealed human head and body lice to be almost genetically identical; although con-specific, they nevertheless occupy distinct ecological niches and have differing feeding patterns. Most importantly, while head lice are not known to be vector competent, body lice can transmit three serious bacterial diseases; epidemictyphus, trench fever, and relapsing fever. In order to gain insights into the molecular bases for these differences, we analyzed alternative splicing (AS) using next-generation sequencing data for one strain of head lice and one strain of body lice. We identified a total of 3,598 AS events which were head or body lice specific. Exon skipping AS events were overrepresented among both head and body lice, whereas intron retention events were underrepresented in both. However, both the enrichment of exon skipping and the underrepresentation of intron retention are significantly stronger in body lice compared with head lice. Genes containing body louse-specific AS events were found to be significantly enriched for functions associated with development of the nervous system, salivary gland, trachea, and ovarian follicle cells, as well as regulation of transcription. In contrast, no functional categories were overrepresented among genes with head louse-specific AS events. Together, our results constitute the first evidence for transcript pool differences in head and body lice, providing insights into molecular adaptations that enabled human lice to adapt to clothing, and representing a powerful illustration of the pivotal role AS can play in functional adaptation.


[Biomarkers of endocrine therapy-resistance in breast cancers].
During the past decade, molecular signatures allowed a better classification of breast carcinoma and a better evaluation of their prognosis. However, we still need predictive factors of treatment and /or prognosis factors specific of each patient. Regarding hormonal therapy, expression of hormone receptors is essential, but not sufficient to accurately predict response to treatment for all patients. To date, numerous data have identified different pathways of resistance to hormonal treatment, associated with heterogeneity in response to long term treatment. The challenge remains to identify and to anticipate these changes for each patient.


Can artificial soil be used in the vegetative vigor test for U.S. pesticide registration?
Current testing guidelines for pesticide registration for the protection of nontarget plants calls for the use of sterilized, standardized soil consisting of primarily sandy loam, loamy sand, loamy clay, or clay loam that contains up to 3% organic matter. Low organic matter soils can be difficult to manage in a greenhouse setting because when soils dry, they contract, causing impeded water infiltration, or when overwatered, poor drainage increases the chances of anaerobic conditions. The purpose of this study was to determine if the results for the vegetative vigor test differed when using either natural or artificial soils. The herbicide sulfometuron methyl was applied 14 d after emergence at 0.1 and 0.0032 of the suggested field application rate. Six plant species were tested, 4 of the common test species, Zea mays (corn), Glycine max (soybean), Avena sativa (oat), and Lactuca sativa (lettuce), and 2 native plants of the Willamette Valley, Oregon prairie, Bromus carinatus (California brome) and Ranunculus occidentalis (western buttercup). Herbicide application rate was the most significant factor in the experiment regardless of soil type. The different soils generally produced different results, even though the 2 native soils, one from Oregon and the other from Maryland, are both acceptable soils for the pesticide registration tests. The plants grown on artificial soil produced results generally between the Oregon and Maryland soil results. This study indicates that artificial soils may produce results similar to or more sensitive than soils currently used in the vegetative vigor test.


Sensorimotor Difficulties Are Associated with the Severity of Autism Spectrum Conditions.
Present diagnostic criteria for autism spectrum conditions (ASC) include social communication and interaction difficulties, repetitive behavior and movement, and atypical sensory responsivity. Few studies have explored the influence of motor coordination and sensory responsivity on severity of ASC symptoms. In the current study, we explore whether sensory responsivity and motor coordination differences can account for the severity of autistic behaviors in children with ASC. Thirty-six children participated: 18 (13 male, 5 female) with ASC (ages 7-16: mean age = 9.93 years) and 18 (7 male, 11 female) typically developing (TD) children (ages 6-12; mean age = 9.16 years). Both groups completed a battery of assessments that included motor coordination, sensory responsivity, receptive language, non-verbal reasoning and social communication measures. Children with ASC also completed the Autism Diagnostic Observation Schedule (ADOS) and Autism Diagnostic Interview-Revised (ADI-R). Results showed that children with ASC scored significantly lower on receptive language, coordination, sensory responsivity and a sensorimotor subscale, Modulation of Activity (MoA) compared to the TD group. In the ASC group, MoA significantly predicted ASC severity across all ASC measures; receptive language and sensory responsivity significantly predicted parental reported autism measures; and coordination significantly predicted examiner observed reported scores. Additionally, specific associations were found between the somatosensory perceptive modalities and ASC severity. The results show that sensorimotor skills are associated with severity of ASC symptoms; furthering the need to research sensorimotor integration in ASC and also implying that diagnosis of ASC should also include the assessment of both coordination deficit and atypical sensory responsivity.


Calcium and polyphosphoinositides: their distribution in relation to the membrane changes occurring in the head of boar spermatozoa.
Ejaculated boar spermatozoa, previously incubated in a rigorously Ca++-free medium, were exposed to Ca++ for different incubation times and processed for the detection of Ca++ localization by a pyroantimonate technique. The distribution of polyphosphoinositides, anionic phospholipids natural constituents of membrane known to bind Ca++, was investigated using a specific cytochemical probe, i.e., neomycin conjugated with horseradish peroxidase. The in situ localizations thus obtained revealed: short exposure to Ca++ ions (10 min) evocated a Ca++-induced release of calcium from the nonmitochondrial intracellular store, i.e., the outer acrosomal membrane; a more prolonged exposure (20 min) triggered the occurrence of fusional and exocytotic events, that appeared to be morphologically related to the acrosome reaction; the outer acrosomal membrane, which is the fusigenic sperm membrane, was the elective site of the neomycin/peroxidase labeling. When assayed for the presence of a phospholipase C-like activity, the detergent extract obtained from boar spermatozoa exhibited substantial amount of p-nitrophenyl-phosphorylcholine hydrolyzing activity. The results, on the whole, allow us to suggest a relationship between Ca++ and polyphosphoinositides turnover in the events triggering the acrosome reaction, the exocytotic process peculiar to mammalian spermatozoa.


Early anthropogenic use of hematite on Aurignacian ivory personal ornaments from Hohle Fels and Vogelherd caves, Germany.
The Aurignacian (ca. 43-35 ka) of southwestern Germany is well known for yielding some of the oldest artifacts related to symbolic behaviors, including examples of figurative art, musical instruments, and personal ornaments. Another aspect of these behaviors is the presence of numerous pieces of iron oxide (ocher); however, these are comparatively understudied, likely owing to the lack of painted artifacts from this region and time period. Several Aurignacian-aged carved ivory personal ornaments from the sites of Hohle Fels and Vogelherd contain traces of what appear to be red ocher residues. We analyzed these beads using a combination of macroanalytical and microanalytical methods, including scanning electron microscopy equipped with energy dispersive spectroscopy and Raman spectroscopy. We found that the residue is composed of the iron oxide mineral hematite (Fe2O3). Further analyses on associated archaeological sediments by X-ray diffraction revealed the absence of hematite and other iron oxide mineral phases, suggesting that the hematite residues were intentionally applied to the ivory personal ornaments by human agents. These findings have important implications as they represent evidence for the direct application of ocher on portable symbolic objects by early Homo sapiens in Europe. Furthermore, our results reveal shared behavioral practices from two key Aurignacian sites maintained over several millennia and illuminate aspects of pigment use and symbolic practices during a pivotal time in the cultural evolution of humans.


Patterns in mammalian muscle energetics.
A description of cellular energetics of muscular contraction is given in terms of the rates and extents of high-energy phosphate splitting during contractile activity, in terms of high-energy phosphate resynthesis by respiration and net anaerobic glycolysis, and in terms of the associated uptake and/or release of H+. These chemical changes have been studied quantitatively by rapid freeze-clamping methods and by 31P-NMR methods. The pattern of chemical changes in a fast-twitch glycolytic muscle is rapid depletion of phosphocreatine and later ATP levels, cellular acidification, and a much slower rate of resynthesis of high-energy phosphate compounds during the recovery period afterwards than occurs in the slow-twitch oxidative muscles. In steady-state contractile activity below the maximal, graded levels of high-energy phosphates and of cellular respiration are achieved in both fast-twitch and slow-twitch muscles. Within the metabolic range up to the maximal aerobic capacity, which differs several-fold for different fibre types, this gradation is mediated by the creatine kinase reaction and phosphocreatine stores. Thus while the amount of enzyme present and the content of phosphocreatine differs among muscles of different types, the same general energetic function is seen to occur in all muscle cells. The creatine kinase reaction is both an energy reservoir and a buffer preventing large swings in the ATP/ADP ratios.


Metabonomics uncovers a reversible proatherogenic lipid profile during infliximab therapy of inflammatory bowel disease.
One-third of inflammatory bowel disease (IBD) patients show no response to infliximab (IFX) induction therapy, and approximately half of patients responding become unresponsive over time. Thus, identification of potential treatment response biomarkers are of great clinical significance. This study employs spectroscopy-based metabolic profiling of serum from patients with IBD treated with IFX and healthy subjects (1) to substantiate the use of spectroscopy as a semi-invasive diagnostic tool, (2) to identify potential biomarkers of treatment response and (3) to characterise the metabolic changes during management of patients with tumour necrosis factor-α inhibitors. Successive serum samples collected during IFX induction treatment (weeks 0, 2, 6 and 14) from 87 IBD patients and 37 controls were analysed by 1H nuclear magnetic resonance (NMR) spectroscopy. Data were analysed with principal components analysis and orthogonal projection to latent structures discriminant analysis using SIMCA-P+ v12 and MATLAB. Metabolic profiles were significantly different between active ulcerative colitis and controls, active Crohn's disease and controls, and quiescent Crohn's disease and controls. Metabolites holding differential power belonged primarily to lipids and phospholipids with proatherogenic characteristics and metabolites in the pyruvate metabolism, suggestive of an intense inflammation-driven energy demand. IBD patients not responding to IFX were identified as a potentially distinct group based on their metabolic profile, although no applicable response biomarkers could be singled out in the current setting. 1H NMR spectroscopy of serum samples is a powerful semi-invasive diagnostic tool in flaring IBD. With its use, we provide unique insights into the metabolic changes taking place during induction treatment with IFX. Of distinct clinical relevance is the identification of a reversible proatherogenic lipid profile in IBD patients with active disease, which partially explains the increased risk of cardiovascular disease associated with IBD.


Utilization of the Family Health Program in metropolitan regions: a methodological approach.
To present the methodological approach used to define the profile of health services utilization by the population enrolled in the Family Health Program. Three patterns of services utilization accessed by the population were considered: residual, partial and full. These patterns were identified based on the range of actions of the Family Health Program that are accessed by the population. An enquiry was conducted in 2006 with two-stage sampling in an area characterized by high levels of social exclusion in the city of São Paulo, Brazil. In the first stage, 960 people participating in family health teams were drawn and classified by the community-based health agents as "full use" or not of the health services. In the second stage, 173 drawn subjects were then classified according to the patterns of services utilization. Subjects were classified as full users (16%), partial users (57%) and residual users (26%), and had different social and demographic characteristics. There was a selective and focused utilization of the services offered by the Family Health Program. Being male, having a level of schooling above the fifth grade of elementary school, having a paid job and accessing medical care systems implied lower adhesion to the services, even though the study focused on regions with few options of healthcare services. Even in areas of high social exclusion and low offer of health services, 25% of the enrolled population did not use the services offered at the Family Health Units, receiving only home visits. Methodologies that are capable of capturing distinct patterns of health services utilization by the population may contribute to improve services evaluation.


New-onset versus prior history of atrial fibrillation: Outcomes from the AFFIRM trial.
There are limited data on prognosis and outcomes of patients with new-onset atrial fibrillation (AF) compared with those with a prior history of AF. We conducted a comparison of these 2 groups in the AFFIRM trial. New-onset AF was the qualifying arrhythmia in 1,391 patients (34%). Compared with patients with prior history of AF, patients with new-onset AF were more likely to have a history of heart failure. There was no mortality difference between rate control (RaC) and rhythm control (RhC) among patients with new-onset AF (17% vs 20%, P = .152). In the univariate model, new-onset AF was associated with increased risk of mortality compared with history of prior AF (RaC unadjusted hazard ratio [HR] 1.36 [P = .010], RhC unadjusted HR 1.39 [P = .003]). However, after multivariate adjustments, new-onset AF did not carry an increased risk of mortality (RaC adjusted HR 1.14 [P = .370], RhC adjusted HR 1.16 [P = .248]). Subjects with new-onset AF randomized to the RhC arm were more likely to remain in normal sinus rhythm at follow-up (adjusted HR 0.79, P = .012) compared with patients with prior history of AF. In a multivariable analysis adjusting for confounders, new-onset AF was not associated with increased mortality compared with prior history of AF regardless of the treatment strategy. Patients with new-onset AF treated with the rhythm control strategy were more likely to remain in normal sinus rhythm on follow-up.


Type of delivery and perinatal outcomes in pregnant women diagnosed with COVID-19: A systematic review and meta-analysis.
There is paucity in the literature that provides a comprehensive and evidence-based conclusion regarding this topic. To compare perinatal outcomes of vaginal and caesarean section delivery in women diagnosed with COVID-19 by meta-analysis of literature. The search was conducted on MEDLINE, EMBASE, LILACS, CINAHL, Scopus, Web of Science and Cochrane Database of Systematic Reviews by 25th May 2022. The inclusion criteria involved pregnant women diagnosed with COVID-19 who underwent caesarean section and those who had vaginal deliveries. The included studies were meta-analyzed for various outcomes including: Gestational age, maternal intensive care unit admission, maternal death, prematurity, newborn birth weight, newborn intensive care unit admission, Apgar scores, newborn death, and newborn vertical transmission of COVID-19. The meta-analysis was conducted using Comprehensive Meta-Analysis (CMA) software, version 3.3.070. The meta-analyses included 2,566 deliveries (866 caesarean sections and 1,700 vaginal deliveries) and identified that caesarean section was significantly associated with increased prematurity (OR 2.5 [1.7; 3.6], p < 0.001), lower birth weight (-118 g [-170; -66], p < 0.001), and a higher need for maternal (OR 9.54 [5.22; 17.43], p < 0.01) and neonatal intensive care unit intervention (OR 3.67 [2.71; 4.96], p < 0.01) compared to vaginal delivery. COVID-19 infection alone should not be an indication for caesarean section as there is insufficient evidence that caesarean section reduces mortality, improves birth conditions, or prevents vertical transmission. Additionally, caesarean section is associated with poorer perinatal outcomes compared to vaginal delivery.


Impact of Truncation on Optoelectronic Properties of Azaborole Helicenes.
Herein, we report configurationally stable singly-truncated (ST) and structurally flexible doubly-truncated (DT) helically chiral compounds derived from azabora[7]helicenes by a hypothetical removal of a single or two C=C double bonds. The singly-truncated constitutional isomers were synthesized from either benzoisoquinoline (BIQ) or phenantherene building blocks and the corresponding biaryls in excellent yields to give azabora[5]helicenes with a pendant phenyl ring at a sterically hindered position. These systems highlight the electronic impact of the nitrogen donor substitution position. The compounds with a disrupted BIQ moiety (STN) possess remarkable photoluminescence quantum yields of up to 0.53 in the solid state and a blue emission in solution with dissymmetry factors of up to ca. 3×10-3 . Upon cooling to 79 K all compounds exhibit phosphorescence with lifetimes of up to ca. 0.5 s. A methyl complex of azabora[7]helicene showing excellent configurational stability was used as a chiral inducer embedded in an emissive polymer (F8BT) to produce circularly polarized organic light emitting diodes with an electroluminescence dissymmetry factor gEL of up to 0.54.


[Clinical characteristics of angina pectoris in relation to disease of the coronary arteries].
Clinical and coronary angiographic comparison conducted in 164 patients with typical angina pectoris demonstrated that the nature of the pains may serve as a criterion of the degree of stenosis and the number of diseased arteries. The development of typical anginal seizures indicates the presence of clear changes in the coronary arteries. A daily intake of over 10 pills of nitroglycerine is a sign of a narrowing of the diameter of the coronary arteries exceeding 75%. A growing number of daily taken pills of nitroglycerine indicates an increase in number of diseased coronaries. Pains occurring during or after meals, when lying down, awakening the patient at night are typical for coronary narrowings exceeding 75% of their diameter. Usually such pains are observed in patients with 2 and more cornary arteries affected by the disease. Rightward irradiation and especially strictly localized pains are often associated with the lesion of the right coronary artery.


Immunization with infective larvae of Strongyloides ratti (Nematoda) exposed to microwave radiation.
Microwaves have not been tested previously for possible application in producing immunogenic preparations of parasites. This study examines the immunizing capacity of microwave-irradiated, infective larvae of Strongyloides ratti in rats. Rats were inoculated subcutaneously with untreated, microwaved, or microwaved and homogenized larvae, or distilled water, and challenged with untreated larvae. Data were collected on egg production and worm number/rat during primary infections and on egg production, worm number/rat, worm size, and eggs in utero/worm following challenge. Our results demonstrated that microwaved, infective larvae (intact or homogenized) of S. ratti were immunogenic for rats, even though they were incapable of reaching the intestine and maturing to adult worms. The immunity elicited by exposure to microwaved larvae was characterized on challenge by a significant reduction in the number of eggs produced/worm, by the formation of perioral plugs, and by reductions in worm numbers and size. These results suggest that microwave radiation may provide a valuable new tool for parasitic vaccine production. In addition, we have demonstrated the occurrence of a feature of the immune response of rats to S. ratti that may have been overlooked previously; i.e., a gut-level response that was elicited by larvae, but manifested against adult worms in the intestine.


Genetic data provide evidence for wind-mediated transmission of highly pathogenic avian influenza.
Outbreaks of highly pathogenic avian influenza in poultry can cause severe economic damage and represent a public health threat. Development of efficient containment measures requires an understanding of how these influenza viruses are transmitted between farms. However, the actual mechanisms of interfarm transmission are largely unknown. Dispersal of infectious material by wind has been suggested, but never demonstrated, as a possible cause of transmission between farms. Here we provide statistical evidence that the direction of spread of avian influenza A(H7N7) is correlated with the direction of wind at date of infection. Using detailed genetic and epidemiological data, we found the direction of spread by reconstructing the transmission tree for a large outbreak in the Netherlands in 2003. We conservatively estimate the contribution of a possible wind-mediated mechanism to the total amount of spread during this outbreak to be around 18%.


Current status of human conception in vitro.
The scientific and medical advances culminating in the introduction of in vitro fertilization of human oocytes into clinical practice are reviewed. Current methods that use clomiphene, human menopausal gonadotrophin, and both as follicular stimulants, and the endogenous LH surge or an injection of human chorionic gonadotrophin to induce ovulation are described. The effects of multifolliculation, the diurnal rhythm of the LH surge, and the collection of oocytes from the ovary are related to current clinical practice. The success of in vitro fertilization for infertile men and women is considered in relation to the nature of embryonic growth in vitro. Investigations into blastulation, hatching from the zona pellucida and the use of DNA probes for typing embryos are described. The implantation of embryos is the major remaining problem, and physiological and statistical analyses of implantation are given, comparing results from different clinics. The possibility of embryo 'helping' and factors leading to multipregnancy are considered, and details are given of the incidence of abortion and the birth of children. The use of immature oocytes, the frozen-storage of embryos and methods of raising the chance of implantation are described briefly.


Hypoxia and vascular endothelial growth factor acutely up-regulate angiopoietin-1 and Tie2 mRNA in bovine retinal pericytes.
The angiopoietin/Tie2 system is a predominant regulator of vascular development. This vascular development appears to be controlled and completed by the coordinated actions of two vascular cells, endothelial cells and their surrounding supporting cells, smooth muscle cells, or pericytes. The role of the angiopoietin/Tie2 system has been studied, but these studies are limited mostly to endothelial cells. In this study, using bovine retinal pericytes (BRP), we investigated the effect of two known angiogenic stimuli, hypoxia and vascular endothelial growth factor (VEGF) treatment, on the regulation of the angiopoietin/Tie2 system. Hypoxia (2% O(2) concentration) was acquired by a hypoxia chamber. Both hypoxia and VEGF (10 ng/ml) treatment significantly increased angiopoietin-1 (Ang1) mRNA expression. This marked augmentation occurred acutely (maximal increase at 2 h) and subsequently decreased. In contrast, angiopoietin-2 (Ang2) mRNA expression was unaltered in BRP upon both treatment. Significant up-regulation of Tie2 mRNA expression was found and lasted up to 12 h. However, using bovine aortic endothelial cells (BAEC), we found that only Ang2 expression, but neither Ang1 nor Tie2, responded to these two angiogenic stimuli, which was consistent with many previous reports. In conclusion, our data suggest that both hypoxia and VEGF treatment differentially regulate the angiopoietin/Tie2 system in the two vascular cells and that, particularly in BRP, the regulation of Ang1, but not Ang2, and Tie2 expression may play an important role in vascular development.


Acceptability and tolerability of and adherence to HIV preexposure prophylaxis among Toronto gay and bisexual men: a pilot study.
Preexposure prophylaxis is efficacious at preventing HIV infection, but concerns persist about adherence and sexually transmitted infections (STIs). We assessed preexposure prophylaxis acceptability, adherence and clinical outcomes in a pilot demonstration project. HIV-uninfected adult gay and bisexual men who scored 10 or higher on a validated HIV risk score (HIV Incidence Risk Index for MSM) and reported condomless receptive anal sex were sequentially enrolled into a 1-year open-label single-arm pilot study of daily oral therapy with tenofovir disoproxil fumarate/emtricitabine in Toronto. The primary outcome was acceptability of preexposure prophylaxis. Secondary outcomes were preexposure prophylaxis adherence (4-d recall, pill count and dried blood spot analysis), HIV seroconversion, STIs and adverse events. Of the 86 men screened, 52 were enrolled. Participants were mostly young (median age 33 yr [interquartile range (IQR) 28-37 yr) white (38 [73%]) gay (49 [94%]) men. Preexposure prophylaxis acceptability was high: all participants reported their experience as "good" or "very good." The median adherence rate was high, at 100% (IQR 95%-100%) by self-report and 96.9% (IQR 93.4%-98.4%) by pill count. Dried blood spot analysis suggested that doses were taken 4-7 days/week at 88.7% (173/195) of month 3-12 visits. No cases of HIV seroconversion occurred, but 25 participants (48%) experienced at least 1 bacterial STI, with incidence rates per 100 person-years of 32.8, 32.8, 8.2 and 8.2 for chlamydia, gonorrhea, syphilis and lymphogranuloma venereum, respectively. No adverse events led to discontinuation of prophylaxis, but the estimated glomerular filtration rate declined by 0.22 mL/min per month. Preexposure prophylaxis was associated with high adherence and acceptability and no HIV infections in this study. Frequent STIs and clinically unapparent toxic renal effects reinforce the need for ongoing vigilance. ClinicalTrials. gov, no. NCT02149888.


Acute exposure to long-chain fatty acids impairs {alpha}2-adrenergic receptor-mediated antilipolysis in human adipose tissue.
The acute in vitro and in vivo effects of long-chain fatty acids (LCFAs) on the regulation of adrenergic lipolysis were investigated in human adipose tissue. The effect of a 2 h incubation, without or with LCFA (200 mumol/l), on basal and hormonally induced lipolysis was tested in vitro on isolated fat cells. The lipolytic response to epinephrine was enhanced by suppression of the antilipolytic alpha(2)-adrenergic effect. Then, healthy lean and obese male subjects performed a 45 min exercise bout at 50% of their heart rate reserve either after an overnight fast or 3 h after a high-fat meal (HFM: 95% fat, 5% carbohydrates). Subcutaneous adipose tissue lipolysis was measured by microdialysis in the presence or absence of an alpha-antagonist (phentolamine). In vivo, a HFM increased plasma levels of nonesterified fatty acids in lean and obese subjects. In both groups, the HFM did not alter hormonal responses to exercise. Under fasting conditions, the alpha(2)-adrenergic antilipolytic effect was more pronounced in obese than in lean subjects. The HFM totally suppressed the alpha(2)-adrenergic antilipolytic effect in lean and obese subjects during exercise. LCFAs per se, in vitro as well as in vivo, suppress alpha(2)-adrenergic-mediated antilipolysis in adipose tissue. LCFA-mediated suppression of antilipolytic pathways represents another mechanism whereby a high fat content in the diet might increase adipose tissue lipolysis.


Donnan equilibrium and pH gradient in isolated tracheal apical membrane vesicles.
Apical plasma membrane vesicles isolated from bovine tracheal epithelium were found to possess both an electrical potential gradient (delta psi) and a transmembrane pH gradient (delta pH). The delta psi was calculated from the distribution of 86Rb+ in the presence of the ionophore valinomycin, and delta pH was determined from the distribution of [14C]methylamine under conditions of apparent equilibrium. Maximal values for delta psi of -54.3 +/- 2.5 mV and for delta pH of 0.75 +/- 0.07 pH units were obtained under low ionic strength conditions. Increasing the ionic strength by the addition of 50 mM of either permeant or impermeant electrolytes reduced delta psi to near zero values. The delta pH varied in parallel with the delta psi. The results suggest that both 86Rb+ in the presence of valinomycin and H+ are in a Donnan equilibrium with impermeant negative charge in the vesicle interior.


Living-skin Detection using Multi-layer Skin Property Perceived by the Structured Light.
Living-skin detection has been used to prevent the attack of face fraud in a face recognition system. In this paper, we propose a new concept that exploits the multi-layer structure property of skin for living-skin detection. We observe a significant difference in the blur of the laser spot created by the structured light on the skin and non-skin due to the characteristic properties of laser photons in skin penetration and reflection. Based on this observation, we designed a new living-skin detection algorithm to differentiate skin and non-skin based on the blur detection of laser spots. The experimental results show that the proposed setup and method have a promising performance with an averaged precision of 96.7%, averaged recall of 82.2%, and averaged F1-score of 88.6% on a dataset of 20 adult subjects. This demonstrates the effectiveness of the new concept that uses multi-layer properties of skin tissues for living-skin detection, which may lead to new solutions for face anti-spoofing.


In vitro cytotoxicity of maxillofacial silicone elastomers: effect of accelerated aging.
The purpose of this in vitro study was to evaluate the cytotoxicity of three maxillofacial silicone elastomers at 24, 48, and 72 h on L-929 cells and to determine the effect of accelerated aging on the cytotoxicity of these silicone elastomers. Disc-shaped test samples of maxillofacial silicone elastomers (Cosmesil, Episil, Multisil) were fabricated according to manufacturers' instructions under aseptic conditions. Samples were then divided into three groups: (1) not aged; (2) aged for 150 h with an accelerated weathering tester; and (3) aged for 300 h. Then the samples were placed in Dulbecco's Modified Eagle Medium/Ham's F12 (DMEM/F12) for 24, 48, and 72 h. After the incubation periods, cytotoxicity of the extracts to cultured fibroblasts (L-929) was measured by MTT assay. The degree of cytotoxicity of each sample was determined according to the reference value represented by the cells with a control (culture without sample). Statistical significance was determined by repeated measurement ANOVA (p < 0.01) followed by Duncan's test (p < 0.05). All test materials in each group demonstrated high survival rates in MTT assay (Episil; 93.84%, Multisil; 88.30%, Cosmesil; 87.50%, respectively); however, in all groups, Episil material demonstrated significantly higher cell survival rate after each of the experimental incubation periods (p < 0.05). Accelerated aging for 150 and 300 h had no significant effect on the biocompatibility of maxillofacial silicone elastomers tested (p > 0.05).


Hypo-osmotic Stress Increases Permeability of Individual Barriers in Escherichia coli Cell Envelope, Enabling Rapid Drug Transport.
Survival of foodborne Gram-negative bacteria during osmotic stress often leads to multidrug resistance development. However, despite the concern, how osmoadaptation alters drug penetration across the Gram-negative bacterial cell envelope has remained inconclusive for years. Here, we have investigated drug permeation and accumulation inside hypo-osmotically shocked Escherichia coli. Three different quaternary ammonium compounds (QACs) are used as cationic amine-containing drug representatives; they also serve as envelope permeability indicators in different assays. Propidium iodide fluorescence reveals cytoplasmic accumulation and overall envelope permeability, while crystal violet sorption and second harmonic generation (SHG) spectroscopy reveal periplasmic accumulation and outer membrane permeability. Malachite green sorption and SHG results reveal transport across both the outer and inner membranes and accumulation in the periplasm as well as cytoplasm. The findings are found to be complementary to one another, collectively revealing enhanced permeabilities of both membranes and the periplasmic space in response to hypo-osmotic stress in E. coli. Enhanced permeability leads to faster QACs transport and higher accumulation in subcellular compartments, whereas transport and accumulation both are negligible under isosmotic conditions. The QACs' transport rates are found to be highly influenced by the osmolytes used, where phosphate ion emerges as a key facilitator of transport across the periplasm into the cytoplasm. E. coli is found viable, with morphology unchanged under extreme hypo-osmotic stress; i.e., it adapts to the situation. The outcome shows that the hypo-osmotic shock to E. coli, specifically using phosphate as an osmolyte, can be beneficial for drug delivery.


Oral manifestations of celiac disease: a clinical guide for dentists.
Celiac disease (gluten sensitive enteropathy) is a common disorder affecting both children and adults. As many people with celiac disease do not present with the classic malabsorptive syndrome, delays in diagnosis are common. Dental enamel defects and recurrent aphthous ulcers, which may occur in patients with celiac disease, may be the only manifestation of this disorder. When dentists encounter these features, they should enquire about other clinical symptoms, associated disorders and family history of celiac disease. In suspected cases, the patient or family physician should be advised to obtain serologic screening for celiac disease and, if positive, confirmation of the diagnosis by intestinal biopsy. Dentists can play on important role in identifying people who may have unrecognized celiac disease. Appropriate referral and a timely diagnosis can help prevent serious complications of this disorder.


[Molecular structure of hybrid phage phi80hy43].
The phage hybrid phi80hy43 derived from a vegetative cross phi80cIhlambda x lambdacIc17 was constructed for discrimination phi80mono- and polylysogens. Molecular structure of this hybrid was established using heteroduplex analysis and restriction endonuclease EcoRI. It is found that the hybrid phi80hy43 represents a phage phi80 containing a foreign piece of DNA between genes cI and 0. The length of this piece of DNA comprises 0.7%, corresponding to the length of cy-cII region of th phage lambda. So it is believed that the hybrid phi80hy43 was formed due to insertion of the lambdacy region with the mutation c17 into phi80hlambda phage genome.


Estimating the effects of lockdown timing on COVID-19 cases and deaths in England: A counterfactual modelling study.
During the first wave of the COVID-19 pandemic, the United Kingdom experienced one of the highest per-capita death tolls worldwide. It is debated whether this may partly be explained by the relatively late initiation of voluntary social distancing and mandatory lockdown measures. In this study, we used simulations to estimate the number of cases and deaths that would have occurred in England by 1 June 2020 if these interventions had been implemented one or two weeks earlier, and the impact on the required duration of lockdown. Using official reported data on the number of Pillar 1 lab-confirmed cases of COVID-19 and associated deaths occurring in England from 3 March to 1 June, we modelled: the natural (i.e. observed) growth of cases, and the counterfactual (i.e. hypothetical) growth of cases that would have occurred had measures been implemented one or two weeks earlier. Under each counterfactual condition, we estimated the expected number of deaths and the time required to reach the incidence observed under natural growth on 1 June. Introducing measures one week earlier would have reduced by 74% the number of confirmed COVID-19 cases in England by 1 June, resulting in approximately 21,000 fewer hospital deaths and 34,000 fewer total deaths; the required time spent in full lockdown could also have been halved, from 69 to 35 days. Acting two weeks earlier would have reduced cases by 93%, resulting in between 26,000 and 43,000 fewer deaths. Our modelling supports the claim that the relatively late introduction of social distancing and lockdown measures likely increased the scale, severity, and duration of the first wave of COVID-19 in England. Our results highlight the importance of acting swiftly to minimise the spread of an infectious disease when case numbers are increasing exponentially.


Econazole selectively induces cell death in NF1-homozygous mutant tumor cells.
Cutaneous neurofibromas (cNFs) are tumors that develop in more than 99% of individuals with neurofibromatosis type 1 (NF1). They develop in the dermis and can number in the thousands. cNFs can be itchy and painful and negatively impact self-esteem. There is no US Food and Drug Administration (FDA)-approved drug for their treatment. Here, we screen a library of FDA-approved drugs using a cNF cell model derived from human induced pluripotent stem cells (hiPSCs) generated from an NF1 patient. We engineer an NF1 mutation in the second allele to mimic loss of heterozygosity, differentiate the NF1+/- and NF1-/- hiPSCs into Schwann cell precursors (SCPs), and use them to screen a drug library to assess for inhibition of NF1-/- but not NF1+/- cell proliferation. We identify econazole nitrate as being effective against NF1-/- hiPSC-SCPs. Econazole cream selectively induces apoptosis in Nf1-/- murine nerve root neurosphere cells and human cNF xenografts. This study supports further testing of econazole for cNF treatment.


Two perspectives on the needs of individuals bereaved by suicide.
To qualitatively explore the needs of suicidally bereaved individuals, researchers interviewed 11 suicide bereaved individuals and 4 mental health workers. Common themes of bereaved persons included the suicide grief experience, coping, interpersonal domain, struggle with meaning, self-reflection, and moving forward. Mental health workers emphasized the nature of the helping relationship, the need to emotionally process the grief, the centrality of meaning making, importance of support groups, and the role of individual counseling. These 2 perspectives informed "best practices" for postvention (e.g., understanding the need for meaning-making while recognizing when to move on).


Analysis of accretion and deletion at boundaries in dynamic scenes.
In dynamic scenes, the presence of object boundaries is often signaled by the appearance or disappearance of occluded surfaces over time. Such regions of surface accretion or deletion can be found using matching techniques similar to those used to determine optical flow in an image sequence. Regions in one frame that are not adequately matched by any region in previous frames correspond to accretion. Regions that have no matches in subsequent frames correspond to deletion. In either case, an occlusion boundary is present. Furthermore, by associating accretion or deletion regions with a surface on one side of a boundary, it is possible to determine which side of the boundary is being occluded. This association can be based purely on visual motion-the accretion or deletion region moves with the same image velocity as the remaining visible surface to which it is attached.


Obesity and the microbiome.
Obesity constitutes a significant and rapidly increasing public health challenge and is associated with significant co-morbidities and healthcare costs. Although undoubtedly multifactorial, research over the last decade has demonstrated that the microbes that colonize the human gut may contribute to the development of obesity through roles in polysaccharide breakdown, nutrient absorption, inflammatory responses and gut permeability. Studies have consistently shown that the Firmicutes to Bacteroidetes ratio, in particular, is increased in obesity and reduces with weight loss. In addition, we and others have shown that the methanogenic Archaea may also contribute to altered metabolism and weight gain in the host. However, much remains to be learned about the roles of different gut microbial populations in weight gain and obesity and the underlying mechanisms before we can begin to approach targeted treatments.


Screening for proteins with polyglutamine expansions in autosomal dominant cerebellar ataxias.
Expansion of trinucleotide CAG repeats coding for polyglutamine has been implicated in five neurodegenerative disorders, including spinocerebellar ataxia (SCA) 1 and SCA3 or Machado-Joseph disease (SCA3/MJD), two forms of type I autosomal dominant cerebellar ataxias (ADCA). Using the 1C2 antibody which specifically recognizes large polyglutamine tracts, particularly those that are expanded, we recently reported the detection of proteins with pathological glutamine expansions in lymphoblasts from another form of ADCA type I, SCA2, as well as from patients presenting with the distinct phenotype of ADCA type II. We now have screened a large series of patients with ADCA or isolated cases with cerebellar ataxia, for the presence of proteins with polyglutamine expansions. A 150 kDa SCA2 protein was detected in 16 out of 40 families with ADCA type I. This corresponds to 24% of all ADCA type I families, which is much more frequent than SCA1 in this series of patients (13%). The signal intensity of the SCA2 protein was negatively correlated to age at onset, as expected for an expanded and unstable trinucleotide repeat mutation. The disease segregated with markers closely linked to the SCA2 locus in all identified SCA2 families. In addition, a specific 130 kDa protein, which segregated with the disease, was detected in lymphoblasts of patients from nine families with ADCA type II. It was also visualized in the cerebral cortex of one of the patients, demonstrating its translation in the nervous system. Finally, no new disease-related proteins containing expanded polyglutamine tracts could be detected in lymphoblasts from the remaining patients with ADCA or isolated cases with cerebellar ataxia.


A protocol for quantifying hydrogen evolution by dye-sensitized molecular photocathodes and its implementation for evaluating a new covalent architecture based on an optimized dye-catalyst dyad.
A protocol that combines gas chromatography and a high-sensitivity micro Clark-type electrode is described to quantify hydrogen production across gas and solution phases for systems operating at very low currents such as dye-sensitized H2-evolving photocathodes. Data indicate that a significant fraction of H2 remains in aqueous solution even after several hours of experiments. Using this protocol, re-evaluation of a dye-sensitized H2-evolving photocathode based on a dye-catalyst dyad showed a reproducible 66% increase of the faradaic efficiency compared with previously reported headspace GC measurements [Kaeffer et al., J. Am. Chem. Soc., 2016, 138, 12308-12311]. This dyad was based on an organic push-pull dye where donor and acceptor are separated by one thiophene group. Insertion of a second thiophene group between the donor and acceptor led to a more efficient system with 30% improved faradaic efficiency for H2 evolution.


Neoadjuvant chemotherapy followed by surgery for advanced-stage endometrial cancer.
Data showing the value of neoadjuvant chemotherapy (nact) followed by interval debulking surgery (ids) in the management of advanced-stage serous endometrial carcinoma (eca) are limited; the aim of the present study was to expand the knowledge about that treatment strategy in patients with advanced eca, including endometrioid eca. Data were collected retrospectively from all patients with advanced-stage eca treated with nact between 2005 and 2014 at 3 oncology referral centres. Primary outcomes were the radiologic response to nact and achievement of optimal or complete ids. Secondary outcomes were recurrence rate and progression-free and overall survival. Of 102 eca cases included, a complete radiologic response was achieved in only 4 cases, with a partial response being achieved in 72% (64% of endometrioid cases, 80% of serous cases). Complete ids was achieved in 62% of the endometrioid cases and in 56% of the serous eca cases, with optimal ids achieved in 31% and 28% of those cases respectively. Survival rates were calculated for all patients with complete and optimal ids; recurrence was observed in 56% and 67% of the cases respectively, and progression-free survival was 18 months and 13 months respectively. Median survival duration was 24 months for endometrioid eca and 28 months for serous eca. For patients with advanced eca who are not suitable for primary debulking, nact followed by ids can be considered regardless of histologic subtype. The treatment options for this group of patients are limited and have to be explored.


Allegations of sexual misconduct against physicians: a lawyer's perspective.
This article provides recommendations to physicians about avoiding claims of sexual misconduct. No physician is immune to such a claim, and the consequences can be significant. The article outlines these consequences, including law enforcement and Department of Professional Regulation action, media attention, criminal and civil proceedings, and litigation with the physician's insurance company. Recommendations on avoiding claims of sexual misconduct include chaperoned patient examinations, enhanced physician-patient communication, and thorough charting in the patient's record.


Gastric diverticulum: a contemporary review and update in management.
Gastric diverticula (GD) are the rarest form of gastrointestinal tract diverticulum, with an estimated incidence of 0.013-2.6%. GD are poorly understood and there are no established management guidelines. Only sparse updates have been published since the mid-20th century. This paper reviews the current literature and provides some suggested guidelines for the management of GD. A search of Medline via OvidSP and Google Scholar for 'gastric diverticulum' and associated synonyms from the year 1950 onwards was performed. We included randomized controlled trials (RCTs), cohort and case-control studies, and case series. Full text, English language manuscripts on adult populations were included. A total of 103 manuscripts were included in the final selection - 77 individual case studies, 23 case series and three reviews. No RCTs, cohort or case-control studies were found. The case studies represent 305 patients, 50.8% female with average age 49.2 years (range 18-80). The most common symptom was abdominal pain (48.2%). The average maximum diameter was 3.97 cm (range 0.5-9). One hundred and four patients were managed operatively. Despite persistent recommendations in the literature that GD > 4 cm should be considered for resection, there are no data supporting this approach. The evidence pertaining to the management of GD is sparse. The decision for operative management should be individualized and based primarily on the presence of symptoms or complications which may be directly attributable to the GD. Where surgery is indicated, a laparoscopic approach, potentially with intra-operative gastroscopy, is appropriate.


Kinship and similarity in residency status structure female social networks in black-and-white colobus monkeys (Colobus vellerosus).
Kinship shapes female social networks in many primate populations in which females remain in their natal group to breed. In contrast, it is unclear to which extent kinship affects the social networks in populations with female dispersal. Female Colobus vellerosus show routine facultative dispersal (i.e., some females remain philopatric and others disperse). This dispersal pattern allowed us to evaluate if facultative dispersed females form social networks shaped by an attraction to kin, to social partners with a high resource holding potential, or to similar social partners in terms of maturational stage, dominance rank, and residency status. During 2008 and 2009, we collected behavioral data via focal and ad libitum sampling of 61 females residing in eight groups at Boabeng-Fiema, Ghana. We determined kinship based on partial pedigrees and genotypes at 17 short tandem repeat loci. Kinship influenced coalition and affiliation networks in three groups consisting of long-term resident females with access to a relatively high number of female kin. In contrast, similar residency status was more important than kinship in structuring the affiliation network in one of two groups that contained recent female immigrants. In populations with female dispersal, the occurrence of kin structured social networks may not only depend on the kin composition of groups but also on how long the female kin have resided together. We found no consistent support for females biasing affiliation toward partners with high resource holding potential, possibly due to low levels of contest competition and small inter-individual differences in resource holding potential.


Hepatorenal syndrome.
Hepatorenal syndrome (HRS) is a common complication of advanced cirrhosis, characterised by renal failure and major disturbances in circulatory function. Renal failure is caused by intense vasoconstriction of the renal circulation. The syndrome is probably the final consequence of extreme underfilling of the arterial circulation secondary to arterial vasodilatation in the splanchnic vascular bed. As well as the renal circulation, most extrasplanchnic vascular beds are vasoconstricted. The diagnosis of HRS is currently based on the exclusion of other causes of renal failure. The prognosis is very poor, particularly when there is rapidly progressive renal failure (type 1). Liver transplantation is the best option in patients without contraindications to the procedure, but it is not always possible owing to the short survival expectancy. Therapies introduced during the past few years, such as vasoconstrictor drugs (vasopressin analogues, alpha-adrenergic agonists) or the transjugular intrahepatic portosystemic shunt, are effective in improving renal function. Nevertheless, liver transplantation should still be done in suitable patients even after improvement of renal function because the outcome of HRS is poor. Finally, recent findings suggest that the risk of developing HRS in the setting of spontaneous bacterial peritonitis may be reduced by the administration of albumin together with antibiotic therapy, and that of HRS occurring in severe alcoholic hepatitis can be lowered by administration of pentoxifylline. Although these findings need to be confirmed, these two strategies represent innovative approaches to lower the frequency of HRS in clinical practice.


Validating retinal fundus image analysis algorithms: issues and a proposal.
This paper concerns the validation of automatic retinal image analysis (ARIA) algorithms. For reasons of space and consistency, we concentrate on the validation of algorithms processing color fundus camera images, currently the largest section of the ARIA literature. We sketch the context (imaging instruments and target tasks) of ARIA validation, summarizing the main image analysis and validation techniques. We then present a list of recommendations focusing on the creation of large repositories of test data created by international consortia, easily accessible via moderated Web sites, including multicenter annotations by multiple experts, specific to clinical tasks, and capable of running submitted software automatically on the data stored, with clear and widely agreed-on performance criteria, to provide a fair comparison.


Ancestry and phenotype predictions from touch DNA using massively parallel sequencing.
Direct PCR can be used to successfully generate full STR profiles from DNA present on the surface of objects. STR profiles are only of use in cases where a potential donor profile is available for comparison, and DNA is of sufficient DNA quality and quantity to generate a reliable profile. Often, no donor information is available and only trace DNA is present on items. As a result, alternative techniques are required to generate genetic data that can provide investigative leads. Massively parallel sequencing (MPS) offers the ability to detect trace levels of DNA and improve DNA analysis success from touched items. Here, we present the first application of direct PCR coupled with MPS to generate forensic intelligence SNP data from latent DNA. The panels assessed are (1) the HIrisplex System that targets 24 SNPs to simultaneously predict hair and eye, and (2) the Precision ID Ancestry Panel that targets 165 autosomal SNPs indicative of biogeographic ancestry. For each panel, we analysed 60 touched samples across five individuals and four substrates (glass slide, fuse, zip-lock bag and wire) using Ion AmpliSeq Library Preparation Kit on the automated Ion Chef System and Ion Torrent PGM. We examine the SNP recovery, concordance with reference samples and the genotype reproducibility from different substrates and donors. The results demonstrate the application of this approach for obtaining informative genetic from trace amounts of DNA.


Dual-mode operation of 2D material-base hot electron transistors.
Vertical hot electron transistors incorporating atomically-thin 2D materials, such as graphene or MoS2, in the base region have been proposed and demonstrated in the development of electronic and optoelectronic applications. To the best of our knowledge, all previous 2D material-base hot electron transistors only considered applying a positive collector-base potential (VCB > 0) as is necessary for the typical unipolar hot-electron transistor behavior. Here we demonstrate a novel functionality, specifically a dual-mode operation, in our 2D material-base hot electron transistors (e.g. with either graphene or MoS2 in the base region) with the application of a negative collector-base potential (VCB < 0). That is, our 2D material-base hot electron transistors can operate in either a hot-electron or a reverse-current dominating mode depending upon the particular polarity of VCB. Furthermore, these devices operate at room temperature and their current gains can be dynamically tuned by varying VCB. We anticipate our multi-functional dual-mode transistors will pave the way towards the realization of novel flexible 2D material-based high-density and low-energy hot-carrier electronic applications.


Protective effects of zinc-bearing clinoptilolite on broilers challenged with Salmonella pullorum.
This study was conducted to evaluate the effects of zinc-bearing clinoptilolite (ZnCP) on broilers challenged with Salmonella pullorum for a 21-d feeding trial. A total of 240 one-day-old chickens was allocated into 4 treatment groups: 1) nonchallenged control (CON) fed basal diet; 2) S. pullorum-challenged control (SCC) fed basal diet; 3) Salmonella challenge fed basal diet plus antibiotic (ANT); 4) Salmonella challenge fed basal diet plus 0.2% ZnCP (ZnCP). Compared with CON, Salmonella challenge caused adverse effects on ADG (P<0.05), ADFI, feed/gain ratio in broilers, Salmonella and Lactobacillus colonies at 7 d (P<0.05) and 21 d, and serum diamine oxidase (DAO) activity. Moreover, the relative weight of spleen of 21-d birds, malondialdehyde (MDA) content of jejunual and ileal mucosa at 21 d, and the superoxide dismutase (SOD) activity of ileal mucosa were also significantly negatively affected by SCC treatment (P<0.05). Supplementation of ZnCP in basal diet significantly improved ADG and reduced Salmonella colonies in cecal contents and serum DAO activity both at 7 and 21 d (P<0.05), compared with SCC. Furthermore, supplementation with ZnCP as well as ANT significantly decreased the relative weight of spleen at 21 d, reduced MDA contents of jejunual and ileal mucosa at 21 d (P<0.05), and improved the SOD activity of ileal mucosa at 7 d as compared with SCC (P<0.05). The results suggested that ZnCP addition in feed exerted protective effects on performance and gut health of broilers against S. pullorum infection.


The Era of Cerebellar Therapy.
Major advances in our understanding of the neurology/pathology, anatomy/physiology, and molecular biology of the cerebellum have opened a new door for cerebellar ataxias (CAs). We have now entered in the 'era of therapies'. Cures are knocking at the door. We discuss the hot topics in the therapeutic protocols available for CAs, including aminopyridines, noninvasive cerebellar stimulation, anti-oxidant drugs and therapies for immune-mediated cerbellar ataxias (IMCAs), topics emphasized in this issue. The history of the cerebellum is a typical example of the importance of apparently divergent and multi-disciplinary approaches.


A Comparative Study of Target Engagement Assays for HDAC1 Inhibitor Profiling.
Histone deacetylases (HDACs) are epigenetic modulators linked to diseases including cancer and neurodegeneration. Given their therapeutic potential, highly sensitive biochemical and cell-based profiling technologies have been developed to discover small-molecule HDAC inhibitors. Ultimately, the therapeutic action of these inhibitors is dependent on a physical engagement with their intended targets in cellular and tissue environments. Confirming target engagement in the cellular environment is particularly relevant for HDACs since they function as part of cell type-specific multiprotein complexes. Here we implemented two recently developed high-throughput target engagement technologies, NanoBRET and SplitLuc CETSA, to profile 349 compounds in the Epigenetic-Focused collection for HDAC1 binding. We found that the two HDAC1 target engagement assays correlated well with each other and with orthogonal activity-based assays, in particular those carried out in cellular environments rather than with isolated HDAC proteins. The assays detected a majority of the previously described HDAC1 inhibitors in the collection and, importantly, triaged HDAC inhibitors known to target other HDACs.


Branhamella catarrhalis activates human B lymphocytes following interactions with surface IgD and class I major histocompatibility complex antigens.
Branhamella catarrhalis initiated DNA synthesis in human blood or spleen cells enriched for B lymphocytes but did not activate T-lymphocyte-enriched fractions. Monoclonal antibodies were used to determine which B-cell surface molecules were of importance for the activation signal. The addition of monoclonal antibodies reactive with IgD, HLA class I antigens, and B2-microglobulin to B lymphocyte cultures selectively inhibited the B-lymphocyte response to B. catarrhalis. Antibody binding to IgD and class I antigens did not inhibit B-cell proliferation following stimulation with anti-IgM beads, Staphylococcus aureus, or Epstein-Barr virus. This suggests that surface IgD is of major importance for B-lymphocyte stimulation by B. catarrhalis. Since B. catarrhalis binds HLA-ABC containing liposomes it is suggested that a similar binding of B. catarrhalis to HLA-ABC on the surface of B lymphocytes serves as an accessory factor that stabilizes the binding of B. catarrhalis to surface IgD. Activation of human B lymphocytes by B. catarrhalis resulted in changes of cell surface molecules that were quantitatively and qualitatively similar to those that resulted from the activation by S. aureus. Therefore although these two bacteria appear to activate B cells in a similar manner, they induce B-cell proliferation through interactions with different cell surface structures.


Parental age and incidence of cleft lip and cleft palate anomalies.
Maternal and paternal age effects have been studied on 90 patients (61 males and 29 females) suffering from cleft lip, cleft palate and, cleft lip and palate anomalies. Younger mothers are at higher risk of getting a child with congenital oral clefts than the older (above 30 years of age). Parental age gaps increase the incidence of congenital malformations.


EMS and emergency department physician triage: injury severity in trauma patients transported by helicopter.
Many trauma patients who are not severely injured arrive at trauma centers via helicopter emergency medical services (HEMS). To compare the injury severity of patients sent to trauma centers by HEMS from community emergency departments (EDs) with the injury severity of those triaged by prehospital providers to HEMS directly from accident scenes. All records were reviewed from trauma-related missions during 1997for a single HEMS system, extracting information on location, time of day, patient age and gender, mechanism of injury, initial vital signs, Revised Trauma Score (RTS), and the extent of care required during transport. These records were then matched with outcome information routinely supplied to the HEMS system by affiliated trauma centers. Information from patients flown directly from scenes was then compared with that for patients flown from community EDs. Information was obtained for 658 patients flown from scenes and 345 flown from community EDs. There were similar proportions of patients in the two groups, with Injury Severity Scale (ISS) scores less than 6 (11.0% vs. 13.5%), between 6 and 14 (47.0% vs. 49.3%), and greater than 15 (42.0% vs. 37.1%); these were not statistically different (p > 0.05). There was also no significant difference between the groups in the RTS, mean ISS score, intensive care unit length of stay, hospital length of stay, or disposition. Scene and interhospital HEMS trauma missions in this system involve patients of similar injury severities. Prehospital providers may triage trauma patients to HEMS transport with proficiency similar to that of community ED physicians.


Segmental and tandem chromosome duplications led to divergent evolution of the chalcone synthase gene family in Phalaenopsis orchids.
Orchidaceae is a large plant family, and its extraordinary adaptations may have guaranteed its evolutionary success. Flavonoids are a group of secondary metabolites that mediate plant acclimation to challenge environments. Chalcone synthase (CHS) catalyses the initial step in the flavonoid biosynthetic pathway. This is the first chromosome-level investigation of the CHS gene family in Phalaenopsis aphrodite and was conducted to elucidate if divergence of this gene family is associated with chromosome evolution. Complete CHS genes were identified from our whole-genome sequencing data sets and their gene expression profiles were obtained from our transcriptomic data sets. Fluorescence in situ hybridization (FISH) was conducted to position five CHS genes to high-resolution pachytene chromosomes. The five Phalaenopsis CHS genes can be classified into three groups, PaCHS1, PaCHS2 and the tandemly arrayed three-gene cluster, which diverged earlier than those of the orchid genera and species. Additionally, pachytene chromosome-based FISH mapping showed that the three groups of CHS genes are localized on three distinct chromosomes. Moreover, an expression analysis of RNA sequencing revealed that the five CHS genes had highly differentiated expression patterns and its expression pattern-based clustering showed high correlations between sequence divergences and chromosomal localizations of the CHS gene family in P. aphrodite. Based on their phylogenetic relationships, expression clustering analysis and chromosomal distributions of the five paralogous PaCHS genes, we proposed that expansion of this gene family in P. aphrodite occurred through segmental duplications, followed by tandem duplications. These findings provide information for further studies of CHS functions and regulations, and shed light on the divergence of an important gene family in orchids.


Liquid phase condensation in cell physiology and disease.
Phase transitions are ubiquitous in nonliving matter, and recent discoveries have shown that they also play a key role within living cells. Intracellular liquid-liquid phase separation is thought to drive the formation of condensed liquid-like droplets of protein, RNA, and other biomolecules, which form in the absence of a delimiting membrane. Recent studies have elucidated many aspects of the molecular interactions underlying the formation of these remarkable and ubiquitous droplets and the way in which such interactions dictate their material properties, composition, and phase behavior. Here, we review these exciting developments and highlight key remaining challenges, particularly the ability of liquid condensates to both facilitate and respond to biological function and how their metastability may underlie devastating protein aggregation diseases.


Similar effects of selective laser trabeculoplasty and prostaglandin analogs on the permeability of cultured Schlemm canal cells.
To evaluate whether selective laser trabeculoplasty and prostaglandin analogs regulate the permeability of cultured human Schlemm canal cells by inducing intercellular junction disassembly. Laboratory investigation. Intercellular junctions were made visible in living cells by making them fluoresce after transfection with a plasmid expressing the zonula occludens 1 protein tagged with green fluorescent protein. Schlemm canal cells were treated by direct laser irradiation; by exposure to media conditioned by either lasered Schlemm canal cells or trabecular meshwork cells; by exposure to the prostaglandin analogs latanoprost, bimatoprost, and travoprost; or by the addition of the nonprostaglandin agents brimonidine, timolol, and dorzolamide. Junction disassembly was monitored using fluorescence microscopy, and permeability alterations were measured as changes in conductivity using flow meters. The direct laser irradiation of Schlemm canal cells caused a 3-fold increase in conductivity. Exposure of the cells to media conditioned by lasered Schlemm canal cells or trabecular meshwork cells induced junction disassembly and a 2- to 4-fold increase in conductivity. Exposure to prostaglandin analogs also induced junction disassembly and a 4- to 16-fold increase in conductivity, whereas the 3 nonprostaglandin agents tested were ineffective in both regards. Exposure to factors secreted by lasered Schlemm canal cells and lasered trabecular meshwork cells and the application of prostaglandin analogs induced junction disassembly while increasing the permeability of Schlemm canal cells. These findings support our hypothesis that selective laser trabeculoplasty and prostaglandin analogs share a common mechanism that likely mediates their pressure-lowering effects.


[Clinical, epidemiological and treatment aspects of 10 cases of saltwater stingray envenomation].
Stingrays are cartilaginous elasmobranches fishes that can cause severe and potentially fatal injuries in humans. A descriptive and prospective survey was conducted to analyze epidemiological, clinical and treatment aspects of injuries caused by marine stingrays in Adicora, Paraguaná peninsula, Falcon State, a northwestern, semiarid region of Venezuela. Between December 2006 and April 2007, patients with saltwater stingray injuries, attended in the ambulatory emergency service of Adicora, were clinically examined and interrogated. Treatment consisted on the application of a topical antiseptic solution (povidone-iodine), intravenous administration of an anti-inflammatory analgesic, antibiotic therapy per os and tetanus immunization. Ten cases of injures caused by the marine stingray Dasyatis guttata (Dasyatidae) were recorded. 6 (60%) of the envenomation cases occurred in males and 4 (40%) in females, being 8 (80%) adults between 20 and 64 years-old and 2 (2%) above that age. Most of the accidents occurred at December and April holidays (90%) and during the late part of the day (80%) when bathing or swimming at or near the beach (70%). The time between ambulatory medical assistance and the accident varied from 5 to 7 minutes, with a mean time of 5.6 +/- 0.84. Injuries were located at lower limbs in all patients with circular lesions of diameters ranging from 1 to 1.53 cm (X: 1.41 +/- 0.22); bleeding was observed in 70% of the cases while 20% showed cellulitis and cutaneous necrosis. Other clinical manifestations were intense local and irradiated pain [Visual analog scale (VAS) = X: 9.5 +/- 0.71)] and erythema (100%). Systemic symptoms included dyspnea (50%) (VAS = X: 5.4 +/- 1.52) and tachycardia (10%). All patients had a satisfactory post-treatment evolution after 20 to 30 minutes (X = 23.1 +/- 3.41) with pain and dyspnea VAS mean values significantly lower (X = 0.55 +/- 0.44; t = 35.5, p = 0.0001; X = 0.1 +/- 0.22; t = 5.57, p = 0.002, respectively). CONCLUSIONS. Injuries caused by the marine stingray D. guttata are a public health problem, appear to have seasonal pattern, and can produce clinical manifestations of significant severity.


[Prodigiozan as an activator of peritoneal macrophages].
The quantitative cytochemical investigation of the prodigiozan effect on the enzymatic activity of the peritoneal macrophages was performed on mice. The drug was administered in a single dose of 150 microgram/kg 24 hours before the specimen collection. Prodigiozan promoted a reliable increase in the activity of the enzymes participating in glycolysis (lactate and cytoplasmic alpha-glycerophosphate dehydrogenases), hexoso-monophosphatic pathway of glucose oxidation (glucoso-6-phosphate dehydrogenase), succinate dehydrogenase, NADP. N-diaphorase and lysosomal enzymes, such as acid phosphatase and non-specific alpha-naphthyl acetate esterase. The changes indicate that activation of the macrophages is one of the significant mechanisms of increasing the host nonspecific resistance with prodigiozan.


Sling Surgery for Male Urinary Incontinence Including Post Prostatectomy Incontinence: A Challenge to the Urologist.
The management of postprostatectomy urinary incontinence (PPI) is still challenging for urologists. In recent decades, various kinds of male sling system have been developed and introduced; however, they have not yet shown as good a result as that of artificial urinary sphincter (AUS). However, a male sling is still in an important position because patients have a high demand for sling implantation, and it can allow the avoidance of the use of mechanical devices like AUS. Recently, the male sling has been widely used in mild-to-moderate PPI patients; however, there are no studies that compare individual devices. Thus, it is hard to directly compare the success rate of operation, and it is impossible to judge which sling system is more excellent. It is expected that many sling options will be available in addition to AUS in the near future with the technological development of various male slings and the accumulation of long-term surgical outcomes. In that in patients with PPI, sling implantation is an option that must be explained rather than an option that need not be explained to them, this review would share the latest outcomes and complications.