Induction of triploidy in the newt Pleurodeles waltlii by heat shock or hydrostatic pressure: interpretation of the different types of ploidy using a chromosomal marker.
Eggs of diploid females of Pleurodeles waltii, fertilized artificially by waltlii sperm, have been submitted to increased temperature of hydrostatic pressure during the first hour of development. Of the resulting viable individuals about 95% were triploid, the remainder diploid or tetraploid. Other types of ploidy have been observed in abnormal embryos (n, 2n/4n, 5n, n/5n, 6n). Some of the treated eggs came from females with a marker chromosome (pericentric inversion). The karyotypes of the animals developing from such eggs confirm that triploidy results from retention of the second polar body and make it possible to interpret the origin of the other types of ploidy. From the results it can be envisaged how gynogenetic animals can be obtained from eggs inseminated by inactivated sperm and shock-treated to restore diploidy.


Parecoxib reduces renal injury in an ischemia/reperfusion model in rats.
To evaluate the effect of parecoxib (an NSAID) on renal function by measuring plasma NGAL (serum neutrophil gelatinase-associated lipocalin) levels in an induced-ischemia rat model. Forty male Wistar rats were randomly assigned to one of four groups: Ischemia (I), Ischemia/parecoxib (IP), No-ischemia (NI), and No-ischemia/parecoxib (NIP). Body weight, mean arterial pressure, heart rate, body temperature, NGAL levels, and renal histology were compared across groups. The Ischemia (I) group, which did not receive parecoxib, showed the highest NGAL levels (p=0.001), while the IP group, which received the medication, had NGAL levels similar to those of the non-ischemic (NI and NIP) groups. Parecoxib resulted in renal protection in this experimental model.


Growth hormone responses to repeated bouts of aerobic exercise with different recovery intervals in cyclists.
To characterise the specific GH responses to repeated bouts of standardised aerobic exercise in amateur competitive cyclists, 6 volunteers (mean age +/- SE: 28.7 +/- 2.3 yr, range: 18-35 yr) performed two consecutive 30-min cycling sessions at 80% of individual maximal oxygen uptake on three occasions with different time interval between bouts: 2 h (EXP A), 4 h (EXP B) and 6 h (EXP C). Serum GH concentration was determined in blood samples collected at 15-min intervals during exercise and following 1 h of recovery. In EXP A and EXP B, peak GH concentration in response to the second bout was significantly lower (p < 0.01) than that of the first bout, but in EXP C no difference was detected between bouts. Similarly, the average integrated GH concentration (AUC), determined during the exercise period and in the following 1 h of recovery in the course of the second bout, was significantly lower than that observed during the first bout only in EXP A (p < 0.05) and EXP B (p < 0.01) and not in EXP C, so that the second bout AUC of EXP C was significantly higher than that of EXP A (p < 0.01) and EXP B (p < 0.01). It was concluded that GH responses to subsequent bouts of aerobic exercise are dependent on the time interval between the exercise sessions.


Direct superoxide anion scavenging by a disodium disuccinate astaxanthin derivative: Relative efficacy of individual stereoisomers versus the statistical mixture of stereoisomers by electron paramagnetic resonance imaging.
Carotenoids are a related group of greater than 600 natural compounds, irrespective of geometric- and stereoisomers, with demonstrated antioxidant efficacy. The carotenoids are broadly divided into "carotenes," or non-oxygen substituted hydrocarbon carotenoids, and "xanthophylls," oxygen-substituted carotenoids. The natural compounds are excellent singlet oxygen quenchers as well as lipid peroxidation chain-breakers; this dual antioxidant capacity is generally attributed to the activity of the polyene chain, and increases with the number of conjugated double bonds along the polyene chain length. However, the poor aqueous solubility of most carotenes and the vast majority of xanthophylls limits their use as aqueous-phase singlet oxygen quenchers and direct radical scavengers. A variety of introduction vehicles (e.g., organic solvents, cyclodextrins) have been used to introduce the insoluble carotenoids into aqueous test systems. Hawaii Biotech, Inc. (HBI) successfully synthesized a novel carotenoid derivative, the disodium disuccinate derivative of astaxanthin (3,3(')-dihydroxy-beta,beta-carotene-4,4(')-dione) in all-trans (all-E) form. The novel derivative is a water-dispersible symmetric chiral molecule with two chiral centers, yielding four stereoisomeric forms: 3R,3(')R and 3S,3(')S (enantiomers), and the diastereomeric meso forms (3R,3(')S and 3(')R,3S). The individual stereoisomers were synthesized at high purity (>90% by HPLC) and compared directly for efficacy with the statistical mixture of stereoisomers obtained from the synthesis from the commercial source of astaxanthin (1:2:1 ratio of 3S,3(')S, meso, and 3R,3(')R, respectively). Direct scavenging of superoxide anion was evaluated in a standard in vitro isolated human neutrophil assay by electron paramagnetic resonance (EPR) imaging, employing the spin-trap DEPMPO. Each novel derivative was tested in pure aqueous formulation and in ethanolic formulation shown to completely disaggregate the compounds in solution. In each case, the ethanolic formulation was a more potent scavenging vehicle. No significant differences in scavenging efficiency were noted among the individual stereoisomers and the statistical mixture of stereoisomers, suggesting that the polyene chain alone was responsible for superoxide scavenging. Dose-ranging revealed that the statistical mixture of stereoisomers of the novel derivative, at millimolar (mM) concentrations, could nearly completely eliminate the superoxide anion signal generated in the activated human neutrophil assay. All ethanolic formulations of the novel derivatives exhibited increased scavenging efficiency over equimolar concentrations of non-esterified astaxanthin delivered in a dimethyl sulfoxide (DMSO) vehicle. These novel compounds will likely find utility in applications requiring aqueous delivery of a highly potent direct radical scavenger.


Ethanol tolerance and the variation of plasma membrane composition of yeast floc populations with different size distribution.
The ethanol tolerance of a self-flocculating yeast strain SPSC01 was investigated in an oxygen-limited fed-batch bioreactor. Employing Focused Beam Reflectance Measurement (FBRM) on-line monitoring system, four yeast floc populations with the average size ranging from 100 to 400mum were obtained. It was found that ethanol tolerance increased with the increasing floc size in the 100, 200, and 300mum floc populations, while increasing the average floc size further to 400mum resulted in lower ethanol tolerance. Examination of the membrane composition of different floc populations revealed that the plasma membrane composition of the floc populations was significantly different in the contents of ergosterol, phosphatidylinositol, as well as phospholipid palmitoleic acid. What's more, the plasma membrane of more ethanol tolerant floc population was less permeable when subjected to 15% (v/v) ethanol shock treatment, and the plasma membrane ATPase activities were higher in the floc populations with higher ethanol tolerance. These results indicate that the average size distribution of the floc populations exerted great influence on the physiological status of yeast cells during the ethanol production process, leading to the changes in plasma membrane composition that contributed to improved ethanol tolerance in self-flocculating yeast SPSC01.


The periodate oxidation of sucrose in aqueous N,N-dimethylformamide.
Sucrose has been oxidized with sodium periodate in 0-50% aqueous N,N-dimethylformamide (DMF). In 50% aqueous DMF the reaction is selective for the glucose ring, yielding a dialdehyde. The increased selectivity is not due to conformational factors but is ascribed to the dissociation of water from cyclic periodate ester species which makes the reaction via the acyclic ester on fructose unfavourable.


Understanding the phenomenological approach to research.
In this paper, Christine Hallett considers the ways in which phenomenology may he used to guide nursing research and discusses some of the difficulties and benefits associated with its adoption in interpretive studies.


Implementation of intimate partner violence education for adolescents with spina bifida.
Intimate partner violence (IPV) prevention among adolescent patients with disability is needed, yet rarely discussed in the clinical setting. This study evaluated the feasibility of implementing a brief educational training based on an evidence-based IPV intervention in a pediatric spina bifida clinic. Frequency of IPV discussion was assessed through evaluation of patient feedback and provider surveys. Adolescent patients with spina bifida aged 12-21 completed after visit surveys before (N= 13) and after the provider training (N= 21). Primary outcomes included frequency of provider discussion about IPV and receipt of patient safety cards. Chi-square tests compared patient feedback prior to and two months following the education session. Provider knowledge and attitude changes were assessed with pre-post surveys. More patients discussed IPV with providers following the education session compared to baseline (p= 0.03). Provider feedback, both immediately and at two months after the education session showed increased awareness of IPV, comfort with assessment, disclosure, and referral to resources. The educational intervention increased provider comfort with addressing IPV within a specialty clinical setting. The frequency of IPV communication significantly increased as compared to baseline, patients reported the discussions were beneficial, and providers reported greater comfort discussing IPV and referring patients to resources.


Inclusion of autistic students: promising modalities for supporting a school team.
School staff who work with autistic students are at a high risk of exhaustion. More training and guidance are needed to enable them to respond to these students' needs. The purpose of this study was to design and evaluate a professional development program offered by an occupational therapist to help a team of school staff facilitate the participation of autistic students and their peers. A design-based research approach guided the development and evaluation of the modalities of the program. A case study was conducted in a school with nine school staff members using individual interviews, questionnaires, and a logbook. The program comprised five modalities: regular presence of the occupational therapist, a needs analysis, content focused on the features of an inclusive school, individual coaching, and team coaching. The following elements emerged as particularly helpful for the participants: frequent interactions with the occupational therapist, opportunities to collaborate, personalized support, experimentation of new practices with the occupational therapist, and concrete training content that can benefit all students. The modalities of the program appear promising to help a school team facilitate the participation of all students, including those who are autistic.Implications for rehabilitationOccupational therapists can help school teams to facilitate the participation of autistic students and their peers.A combination of individual and team coaching sessions appears promising in helping school staff adopt practice changes to make their school more inclusive.Using a multi-tiered approach can help therapists collaborate with school staff members to first find solutions that will be applicable with all of their students.


Pseudouridines in rRNA helix 69 play a role in loop stacking interactions.
The (1)H NMR spectra of RNAs representing E. coli 23S rRNA helix 69 with [1,3-(15)N]pseudouridine modification at specific sites reveal unique roles for pseudouridine in stabilizing base-stacking interactions in the hairpin loop region.


Differential expression of genes encoding 1-aminocyclopropane-1-carboxylate synthase in Arabidopsis during hypoxia.
Ethylene plays an essential role in response to hypoxic stress in plants. In most plant species, 1-aminocyclopropane-1-carboxylate synthase (ACS) is the key enzyme that regulates the production of ethylene. We examined the expression of ACS genes in Arabidopsis during hypoxia. Our data showed that the expression of 4 of the 12 Arabidopsis ACS genes, ACS2, ACS6, ACS7, and ACS9, is induced during hypoxia with three distinct patterns. The hypoxic induction of ACS9 is inhibited by aminooxy acetic acid, an inhibitor of ethylene biosynthesis. In addition, the hypoxic induction of ACS9 is also reduced in etr1-1 and ein2-1, two ethylene insensitive mutants in ethylene-signaling pathways, whereas the addition of 1-aminocyclopropane-1-carboxylic acid, a direct precursor of ethylene, does not induce ACS9 under normoxic conditions. These results indicate that ethylene is needed, but not sufficient, for the induction of ACS9 during hypoxia. This pattern of regulation is similar to that of ADH that encodes alcohol dehydrogenase, which we have reported previously. In contrast, the increased ethylene production during hypoxia has an inhibitory effect on ACS2 induction in roots, whereas ethylene has no effect on the hypoxic induction of ACS6 and ACS7. Based on these results, we propose that two signaling pathways are triggered during hypoxia. One pathway leads to the activation of ACS2, ACS6, and ACS7, whereas the other pathway leads to the activation of ADH and ACS9.


Influence of straw-derived humic acid-like substance on the availability of Cd/As in paddy soil and their accumulation in rice grain.
Humic acid amendments have been widely advocated for the remediation of heavy metal-contaminated soil. However, the impacts of straw-derived humic acid-like substances on the remediation of cadmium (Cd) and arsenic (As) co-contaminated paddy soil remain unclear and the potential mechanism required clarification. In this study, we employed a pot experiment and chose a straw-derived humic acid-like substance (BFA) as the amendment with four doses to investigate how BFA affects the availability of Cd and As in soil and their accumulation in rice. The results showed that grain Cd decreased by 25.65-36.03%, while there was no significant change in total As (TAs) with the addition of BFA. The contents of DCB-Fe, DCB-As and DCB-Cd on the root surface decreased by 6.07-40.54% during the whole growth stage. The addition of BFA significantly decreased the pe + pH and enhanced the transformation of crystalline iron oxides (Fed) into amorphous forms (Feo) in the soil. The CaCl2-extractable Cd decreased and the KH2PO4-extractable As increased with the decrease in pe + pH and Fed and the relative increase in Feo. The correlation analysis showed that the decrease in availability of Cd and translocation factor of Cd effectively decreased the grain Cd and the decrease in DCB-Cd may also contribute to decreasing the uptake of Cd by rice. However, the increase in As of roots and shoots might play key roles in restricting the transport of As to rice grains. Consequently, the addition of BFA could effectively reduce the Cd accumulation in rice under flooding conditions, while no risk of As accumulation in rice grain was observed. The present work provides a new perspective for the application of straw-derived humic acid-like substances as amendments on Cd-As co-contaminated soils, which should be advocated as an eco-friendly, economical and effective soil amendment in the future.


A detailed behavioral analysis of the acute motor effects of caffeine in the rat: involvement of adenosine A1 and A2A receptors.
There is no consensus on the contribution of adenosine A(1) and A(2A) receptor blockade to motor-activating effects of caffeine. Our aim was to use a detailed and continuous observational method to compare the motor effects induced by caffeine with those induced by selective A(1) and A(2A) receptor antagonists. The behavioral repertoire induced by systemic administration of caffeine (3, 10, and 30 mg/kg), A(1) receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (CPT; 1.2, 4.8 and 7.2 mg/kg), and A(2A) receptor antagonist 3-(3-hydroxypropyl)-8-(m-methoxystyryl)-7-methyl-1-propargylxanthine phosphate disodium salt (MSX-3; 1, 3, and 10 mg/kg) was analyzed. The effects of pretreatment with the selective A(1) receptor agonist N (6)-cyclopentyladenosine (CPA; 0.1 mg/g) and the selective A(2A) receptor agonist 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxyamidoadenosine (CGS 21680; 0.2 mg/kg) on the pattern of motor activation induced by caffeine, CPT, or MSX-3 were also examined. The pattern of behavioral activation induced by caffeine was better mimicked by CPT than by MSX-3. Coadministration of CPT and MSX-3 gave different results depending on the dose and the type of behavioral response. CPA was more effective at decreasing the activating effects of caffeine and CPT than those of CGS 21680. On the other hand, CGS 21680 was more effective at decreasing the activating effects of MSX-3 than those of caffeine or CPT. Factor analysis revealed a complex three-dimensional behavioral profile for caffeine that was similar to the profile for CPT and was different from the profile for MSX-3. The results indicate a predominant role for A(1) receptors in the motor-activating effects of acutely administered caffeine.


Factors Associated with Ipsilateral Limb Ischemia in Patients Undergoing Femoral Cannulation Extracorporeal Membrane Oxygenation.
Extracorporeal membrane oxygenation (ECMO) is an important life-saving modality for patients with cardiopulmonary failure. Vascular complications, including clinically significant limb ischemia, may occur as a result of femoral artery cannulation for venoarterial (VA) ECMO. This study examines our institutional experience with femoral VA ECMO and the development of ipsilateral limb ischemia. We performed a retrospective review of all consecutive patients undergoing femoral VA ECMO between 2011 and 2016. The primary endpoint was clinical evidence of limb-threatening ischemia. Multivariate logistic regression analysis was used to identify predictors for limb ischemia after cannulation. Between March 2011 and September 2016, 154 patients underwent femoral cannulation for VA ECMO. Overall in-hospital mortality was 59.7%. Clinically significant ipsilateral limb ischemia occurred in 34 (22%) patients; 7 required four-compartment fasciotomy, and 3 of these patients required amputation. On univariate analysis, a history of pulmonary disease, peripheral arterial disease, and stroke or transient ischemic attack was significantly associated with clinical limb ischemia. On multivariate analysis, younger age (odds ratio [OR], 0.96; 95% confidence interval [CI], 0.93-0.99), diabetes (OR, 2.77; 95% CI, 1.08-7.12), pulmonary disease (OR, 3.86; 95% CI, 1.38-10.78), and peripheral arterial disease (OR, 13.68; CI, 2.75-68.01) were associated with limb ischemia. Lack of prophylactic distal perfusion catheter and arterial cannula size were not independently associated with limb ischemia. Femoral ECMO cannulation can be associated with significant limb ischemia necessitating surgical intervention. Younger patients, as well as those with a history of diabetes, pulmonary disease, and peripheral arterial disease, may be at increased risk for this complication.


The interaction between prorenin, renin and the (pro)renin receptor: time to rethink the role in hypertension.
Elevated prorenin levels are seen in diabetics with microvascular disease. The discovery of a receptor capable of binding renin and prorenin [(P)RR] and triggering an intracellular signal in the laboratory setting raised the expectation that prorenin might be directly responsible for these vascular disorders. However, there has been substantial disagreement concerning the signaling properties of renin and prorenin and it has been impossible to inactivate the (P)RR gene in mouse to define its function. Mouse and rat models in which prorenin is highly overexpressed do not demonstrate the glomerulosclerosis typically seen in severe diabetic nephropathy, but do exhibit an increase in blood pressure that is angiotensin II-dependent. (P)RR has been shown to colocalize with other subunits of the vacuolar ATPase in the kidney and heart and to be necessary for Wnt signaling in a renin-independent manner. Although whole-body inactivation of the (P)RR gene is lethal, tissue-specific inactivation results in severe disorders associated with massive cell death. These results do not support a role of direct prorenin or renin signaling through (P)RR in vascular disorders. Rather, they suggest that the main role of (P)RR is as a subunit of the vacuolarATPase complex. Whether or not (P)RR is responsible for the ability of prorenin to generate angiotensin II in tissues has not been resolved.


Chemical induction of gut β-like-cells by combined FoxO1/Notch inhibition as a glucose-lowering treatment for diabetes.
Lifelong insulin replacement remains the mainstay of type 1 diabetes treatment. Genetic FoxO1 ablation promotes enteroendocrine cell (EECs) conversion into glucose-responsive β-like cells. Here, we tested whether chemical FoxO1 inhibitors can generate β-like gut cells. We used Ngn3-or Villin-driven FoxO1 ablation to capture the distinctive developmental effects of FoxO1 on EEC pool. We combined FoxO1 ablation with Notch inhibition to enhance the expansion of EEC pool. We tested the ability of an orally available small molecule of FoxO1 inhibitor, Cpd10, to phenocopy genetic ablation of FoxO1. We evaluated the therapeutic impact of genetic ablation or chemical inhibition of FoxO1 on insulin-deficient diabetes in Ins2Akita/+ mice. Pan-intestinal epithelial FoxO1 ablation expanded the EEC pool, induced β-like cells, and improved glucose tolerance in Ins2Akita/+ mice. This genetic effect was phenocopied by Cpd10. Cpd10 induced β-like cells that released insulin in response to glucose in gut organoids, and this effect was enhanced by the Notch inhibitor, DBZ. In Ins2Akita/+ mice, a five-day course of either Cpd10 or DBZ induced intestinal insulin-immunoreactive β-like cells, lowered glycemia, and increased plasma insulin levels without apparent adverse effects. These results provide proof of principle of gut cell conversion into β-like cells by a small molecule FoxO1 inhibitor, paving the way for clinical applications.


Phosphorylation of doublecortin by protein kinase A orchestrates microtubule and actin dynamics to promote neuronal progenitor cell migration.
Doublecortin (DCX) is a microtubule-associated protein that is specifically expressed in neuronal cells. Genetic mutation of DCX causes lissencephaly disease. Although the abnormal cortical lamination in lissencephaly is thought to be attributable to neuronal cell migration defects, the regulatory mechanisms governing interactions between DCX and cytoskeleton in the migration of neuronal progenitor cells remain obscure. In this study we found that the G(s) and protein kinase A (PKA) signal elicited by pituitary adenylate cyclase-activating polypeptide promotes neuronal progenitor cells migration. Stimulation of G(s)-PKA signaling prevented microtubule bundling and induced the dissociation of DCX from microtubules in cells. PKA phosphorylated DCX at Ser-47, and the phospho-mimicking mutant DCX-S47E promoted cell migration. Activation of PKA and DCX-S47E induced lamellipodium formation. Pituitary adenylate cyclase-activating polypeptide and DCX-S47E stimulated the activation of Rac1, and DCX-S47E interacted with Asef2, a guanine nucleotide exchange factor for Rac1. Our data reveal a dual reciprocal role for DCX phosphorylation in the regulation of microtubule and actin dynamics that is indispensable for proper brain lamination.


Global Trends in Marine Plankton Diversity across Kingdoms of Life.
The ocean is home to myriad small planktonic organisms that underpin the functioning of marine ecosystems. However, their spatial patterns of diversity and the underlying drivers remain poorly known, precluding projections of their responses to global changes. Here we investigate the latitudinal gradients and global predictors of plankton diversity across archaea, bacteria, eukaryotes, and major virus clades using both molecular and imaging data from Tara Oceans. We show a decline of diversity for most planktonic groups toward the poles, mainly driven by decreasing ocean temperatures. Projections into the future suggest that severe warming of the surface ocean by the end of the 21st century could lead to tropicalization of the diversity of most planktonic groups in temperate and polar regions. These changes may have multiple consequences for marine ecosystem functioning and services and are expected to be particularly significant in key areas for carbon sequestration, fisheries, and marine conservation. VIDEO ABSTRACT.


Probing the importance of spacial and conformational domains in captopril analogs for angiotensin converting enzyme activity.
A new synthesis of 4,5-methano-L-prolines and the enzymatic activity of the corresponding N-(3-mercapto-2-R-methyl-propionyl) analogs as inhibitors of angiotensin converting enzyme are described.


Cadmium isotope fractionation of materials derived from various industrial processes.
Our study represents ϵ(114/110) Cd NIST3108 values of materials resulting from anthropogenic activities such as coal burning, smelting, refining, metal coating, and the glass industry. Additionally, primary sources (ore samples, pigment, coal) processed in the industrial premises were studied. Two sphalerites, galena, coal and pigment samples exhibited ϵ(114/110) CdNIST3108 values of 1.0±0.2, 0.2±0.2, 1.3±0.1, -2.3±0.2 and -0.1±0.3, respectively. In general, all studied industrial processes were accompanied by Cd isotope fractionation. Most of the industrial materials studied were clearly distinguishable from the samples used as a primary source based on ϵ(114/110) Cd NIST3108 values. The heaviest ϵ(114/110) CdNIST3108 value of 58.6±0.9 was found for slag resulting from coal combustion, and the lightest ϵ(114/110) CdNIST3108 value of -23±2.5 was observed for waste material after Pb refinement. It is evident that ϵ(114/110) Cd NIST3108 values depend on technological processes, and in case of incomplete Cd transfer from source to final waste material, every industrial activity creates differences in Cd isotope composition. Our results show that Cd isotope analysis is a promising tool to track the origins of industrial waste products.


The microsomal prostaglandin E synthase-1/prostaglandin E2 axis induces recovery from ischaemia via recruitment of regulatory T cells.
Microsomal prostaglandin E synthase-1 (mPGES-1)/prostaglandin E2 (PGE2) induces angiogenesis through the prostaglandin E2 receptor (EP1-4). Among immune cells, regulatory T cells (Tregs), which inhibit immune responses, have been implicated in angiogenesis, and PGE2 is known to modulate the function and differentiation of Tregs. We hypothesized that mPGES-1/PGE2-EP signalling could contribute to recovery from ischaemic conditions by promoting the accumulation of Tregs. Wild-type (WT), mPGES-1-deficient (mPges-1-/-), and EP4 receptor-deficient (Ep4-/-) male mice, 6-8 weeks old, were used. Hindlimb ischaemia was induced by femoral artery ligation. Recovery from ischaemia was suppressed in mPges-1-/- mice and compared with WT mice. The number of accumulated forkhead box protein P3 (FoxP3)+ cells in ischaemic muscle tissue was decreased in mPges-1-/- mice compared with that in WT mice. Expression levels of transforming growth factor-β (TGF-β) and stromal cell derived factor-1 (SDF-1) in ischaemic tissue were also suppressed in mPges-1-/- mice. The number of accumulated FoxP3+ cells and blood flow recovery were suppressed when Tregs were depleted by injecting antibody against folate receptor 4 in WT mice but not in mPges-1-/- mice. Recovery from ischaemia was significantly suppressed in Ep4-/- mice compared with that in WT mice. Furthermore, mRNA levels of Foxp3 and Tgf-β were suppressed in Ep4-/- mice. Moreover, the number of accumulated FoxP3+ cells in ischaemic tissue was diminished in Ep4-/- mice compared with that in Ep4+/+ mice. These findings suggested that mPGES-1/PGE2 induced neovascularization from ischaemia via EP4 by promoting the accumulation of Tregs. Highly selective EP4 agonists could be useful for the treatment of peripheral artery disease.


Socio-demographic factors and reasons associated with delay in breast cancer presentation: a study in Nigerian women.
We evaluated the effects of selected socio-demographic factors on late presentation and reasons why our breast cancer patients delay reporting for treatment. All female breast cancer patients referred to one of the general surgery out-patient clinics of Lagos State University Teaching Hospital between January 2009 and December 2010 were interviewed. Relevant socio-demographic and clinical data were obtained and reasons for patient delay documented. Univariate and multivariate logistic regression analyses were conducted to calculate odd ratio for delay. A total of 201 patients were enrolled. Mean duration of symptoms was 12.12 months (SD ± 5.18). Delay for more than 3 months before initial medical consultation was observed in 164 patients (81.6%). Increased risk of late presentation was associated with single women (OR=2.054), primary level of education (OR=3.059), negative history of benign breast disease (OR=1.648) and pre-menopause (OR=1.861). Ignorance of the nature of illness, belief in spiritual healing, fear of mastectomy and belief in herbal treatment were the leading reasons for delay. Women with higher risk of late presentation should be the target group during interventions aimed at raising breast cancer awareness. Reasons for patient delay should also be addressed.


Species-Specific Evolution of Ebola Virus during Replication in Human and Bat Cells.
Ebola virus (EBOV) causes a severe, often fatal disease in humans and nonhuman primates. Within the past decade, EBOV has caused two large and difficult-to-control outbreaks, one of which recently ended in the Democratic Republic of the Congo. Bats are the likely reservoir of EBOV, but little is known of their relationship with the virus. We perform serial passages of EBOV in human and bat cells and use circular sequencing to compare the short-term evolution of the virus. Virus populations passaged in bat cells have sequence markers indicative of host RNA editing enzyme activity, including evidence for ADAR editing of the EBOV glycoprotein. Multiple regions in the EBOV genome appear to have undergone adaptive evolution when passaged in bat and human cells. Individual mutated viruses are rescued and characterized. Our results provide insight into the host species-specific evolution of EBOV and highlight the adaptive flexibility of the virus.


Carbonylative Polymerization of Epoxides Mediated by Tri-metallic Complexes: A Dual Catalysis Strategy for Synthesis of Biodegradable Polyhydroxyalkanoates.
Polyhydroxyalkanoates (PHAs) are a unique class of commercially manufactured biodegradable polyesters with properties suitable for partially substituting petroleum-based plastics. However, high costs and low volumes of production have restricted their application as commodity materials. In this study, tri-metallic complexes were developed for carbonylative polymerization via a dual catalysis strategy, and 17 products of novel PHAs with up to 38.2 kg mol-1 Mn values were discovered. The polymerization proceeds in a sequential fashion, which entails the carbonylative ring expansion of epoxide to β-lactone and its subsequent ring-opening polymerization that occurs selectively at the O-alkyl bond via carboxylate species. The wide availability and structural diversity of epoxide monomers provide PHAs with various structures, excellent functionalities, and tunable properties. This study represents a rare example of the preparation of PHAs using epoxides and carbon monoxide as raw materials.


High expression of 90K (Mac-2 BP) is associated with poor survival in node-negative breast cancer patients not receiving adjuvant systemic therapies.
90K (Mac-2 BP) expression was evaluated by immunohistochemistry in paraffin-embedded tissue from a consecutive series of lymph-node negative breast cancer patients who did not receive adjuvant systemic treatment. An independent series of patients served as validation set. The association of 90K expression with risk of recurrence and death was examined in survival analyses together with known prognostic factors. High levels of 90K expression (IHC score>8) were observed in 43 (25.3%) of 170 tumors examined. We found elevated risks of distant recurrence and overall mortality in patients with high 90K expression compared with patients with low 90K expression in their tumors. This increase persisted after adjusting for other prognostic factors in multivariate analysis (hazard ratio=4.084; p<0.001 for recurrence; hazard ratio=4.298; p<0.001 for death). These findings were confirmed in the validation set. Therefore, evaluation of 90K expression may be beneficial to identify lymph-node negative breast cancer patients at lower risk of disease recurrence and death.


Time to effective antibiotic administration in adult patients with septic shock: a descriptive analysis.
To determine the median time to antibiotic administration following the onset of septic shock at our institution as well as the appropriateness of empiric therapy, sources of delay in antibiotic administration and the effect of delays on survival. Retrospective health record review of 55 patients with septic shock admitted to the intensive care unit (ICU) between July 1, 2008 and December 31, 2009. Nine-bed adult medical-surgical ICU within a 300-bed community acute care hospital. Median time to antibiotic administration, appropriateness of empiric therapy, sources of delay in antibiotic administration. The median (min,max) time to the initiation of antibiotics was determined to be 1.7 (0,31) hours. Only 34% (19/55) of patients received antibiotics within the recommended one hour. Empiric antibiotic therapy was determined to be appropriate in 91% (50/55) of patients. The median (min,max) time to administration of effective antibiotic therapy tended to be faster in the emergency room [1.1 (0,16) hours] compared to the ICU [2.3 (0,13)]. The median time to antibiotic administration at our institution following the onset of septic shock was longer than the evidence-based guideline recommendations of within one hour.


Imaging plants dynamics in heterogenic environments.
Noninvasive imaging sensors and computer vision approaches are key technologies to quantify plant structure, physiological status, and performance. Today, imaging sensors exploit a wide range of the electromagnetic spectrum, and they can be deployed to measure a growing number of traits, also in heterogenic environments. Recent advances include the possibility to acquire high-resolution spectra by imaging spectroscopy and classify signatures that might be informative of plant development, nutrition, health, and disease. Three-dimensional (3D) reconstruction of surfaces and volume is of particular interest, enabling functional and mechanistic analyses. While taking pictures is relatively easy, quantitative interpretation often remains challenging and requires integrating knowledge of sensor physics, image analysis, and complex traits characterizing plant phenotypes.


Derivation and Implementation of a Protocol in Israel for Organ Donation after Cardio-Circulatory Death.
Strategies aimed at expanding the organ donor pool have been sought, which has resulted in renewed interest in donation after cardio-circulatory death (DCCD), also known as non-heart beating donors (NHBDs). To describe the derivation and implementation of a protocol for DCCD in Israel and report on the results with the first six cases. After receiving approval from an extraordinary ethics committee, Ministry of Health, the steering committee of the National Transplant Center defined and reached consensus on the unique challenges presented by a DCCD program. These protocol included medical aspects (construction of a clinical pathway), social and ethical aspects (presentation of the protocol at a public gathering(, legal/ethical aspects (consent for organ preservation procedures being either implied if the donor had signed an organ donor card or received directly from a surrogate decision maker), and logistical aspects (pilot study confined to kidney retrieval and to four medical centers). Data regarding organ donors and recipients were recorded. The protocol was implemented at four medical centers. Consent for organ donation was received from four of the six potential donors meeting criteria for inclusion, in all cases, from a surrogate decision maker. Of the eight kidneys retrieved, only four were suitable for transplantation, which was carried out successfully for four recipients. Graft function remained normal in all cases in 6-12 months follow-up. The DCCD program was successfully implemented and initial results are encouraging, suggesting that expansion of the program might further aid in decreasing the gap between needs and availability of organs.


Key determinants of success in fecal microbiota transplantation: From microbiome to clinic.
Fecal microbiota transplantation (FMT) has achieved satisfactory results in preventing the recurrence of Clostridioides difficile infection, but these positive outcomes have only been partially replicated in other diseases. Several factors influence FMT success, including those related to donors and recipients (including diversity and specific composition of the gut microbiome, immune system, and host genetics) as well as to working protocols (fecal amount and number of infusions, route of delivery, and adjuvant treatments). Moreover, initial evidence suggests that the clinical success of FMT may be related to the degree of donor microbial engraftment. The application of cutting-edge technologies for microbiome assessment, along with changes in the current vision of fecal transplants, are expected to improve FMT protocols and outcomes. Here, we review the key determinants of FMT success and insights and strategies that will enable a close integration of lab-based and clinical approaches for increasing FMT success.


Cultural respect in midwifery service provision for Aboriginal women: longitudinal follow-up reveals the enduring legacy of targeted program initiatives.
Culturally competent maternity care provision to Aboriginal and Torres Strait Islander women was identified as a priority area by Australia's National Maternity Services Plan in 2011. While midwifery programs responded by including core Indigenous content and community placements in curricula, little is known about whether knowledge learned, and insights gained in response to these initiatives have endured and been applied in clinical practice. This follow-up study explores the impact of a compulsory Indigenous unit and a remote clinical placement on two cohorts of non-Indigenous midwives who were participants in an earlier 2012-14 study. Fourteen non-Indigenous participants who were either students or recent graduates in 2012-14 were located and re-interviewed in 2019-20. In-depth interviews based on a semi-structured interview guide were conducted by telephone or face-to face; recordings were transcribed and thematically analysed using standard qualitative procedures. Exposure to Indigenous content and settings during training had an enduring impact on participants' midwifery practice; most felt better prepared to provide culturally safe care, build respectful relationships and advocate for improved services for Aboriginal women. Despite this positive legacy, they also expressed apprehension about causing offence and recognised their own knowledge deficits with regard to Aboriginal cultural practices. Organisational constraints, including restrictions on the number of family members accompanying a birthing mother were identified as barriers to optimal care; some positive organisational initiatives were also described. This follow-up study provides encouraging evidence that well-designed and delivered Indigenous content and community placement opportunities in midwifery programs can have a lasting impact on service provision to Aboriginal women, contribute to a more informed, empathetic and culturally competent maternity workforce and help catalyse health service changes towards more culturally safe care.


Epidermal nevus syndrome: case report and review of clinical manifestations.
The epidermal nevus syndrome is a disorder characterized by epidermal nevi and associated neurologic, skeletal, and other abnormalities. We cared for a 3-month-old male with multiple epidermal nevi and severe central nervous system involvement.


A special case on the use of ultrasonography for evaluation of the spinal canal and its contents in adults.
In this article we present a 70-year-old man with a history of severe trauma in the upper thoracic spine. This patient presented at our emergency department with a leaking wound in the lower neck after removal of osteosynthetic material. He had undergone a laminectomy of the third thoracic vertebra in the past. As an incidental finding, while examining the region of the lower neck by transcutaneous ultrasonography, we diagnosed a post-traumatic syrinx of the spinal medulla at this level.


Nano-hydroxyapatite reinforced polyhydroxybutyrate composites: a comprehensive study on the structural and in vitro biological properties.
Nanocomposites based on polyhydroxybutyrate (PHB) and hydroxyapatite (HAp) have recently been proposed for application in bone repair and regeneration, but very limited studies have investigated the effect of HAp on the rheological and thermal behavior of PHB. More important, the efficiency of a biomaterial depends greatly on its ability to interact with cells, but little is known about this interaction for this kind of nanocomposite. Hence, this paper dealt with some of the characteristics of solution-casted PHB/HAp nanocomposite films, and tried to explore the effect of HAp nanoparticles on cellular responses. The results showed that both rheological and thermal properties can be tailored by incorporating appropriate amounts of nanoparticles. In vitro studies showed a significant increase in proliferation and differentiation of MC3T3-E1 on nanocomposites compared to the neat polymer. Surface examination indicated that topography and chemistry of surface are important factors influencing cellular processes; while no cell differentiation was found on the neat polymer, nanocomposite with 15 wt.% filler content exhibited a pronounced differentiation resulting from high surface roughness and large amount of exposed HAp. These results suggest that HAp particles play a much more important role in determining the biological performance of PHB than has previously been supposed.


[Short term clinical outcomes of laparoscopic fertility preserving radical hysterectomy in the management of early stage cervical cancer].
To investigate the feasibility and effectiveness of laparoscopic radical trachelectomy and lymphadenectomy in the treatment of early-stage cervical cancer. The clinical data of 6 patients (stage 1a2 to 1b1), who underwent laparoscopic fertility-preserving radical operation for cervical cancer in our department from February 2009 to October 2010, were retrospectively analyzed in terms of operation duration, intra-operative blood loss, postoperative pathology, complications, and pregnancy. Both radical resection of cervical and pelvic lymph node dissection were completed under laparoscopy, and only the cervical and vaginal cuffs were closed from vagina. The operation duration ranged 155-210 min (mean: 185 min) and the intra-operative blood loss was approximately 60-120 ml(mean: 105 ml). The average length of hospital stay was 18 days without complications, postoperative infection, and bleeding. Postoperative pathology showed no lymph node metastasis, and no ligament, blood vessels, vaginal cutting margin, or upper part of cervix was invaded by tumor cells. During the 8-20-month follow-up, 1 patient had become pregnant for 4 months and no case experienced tumor recurrence. Laparoscopic fertility-preserving lymphadenectomy and radical trachelectomy is feasible for patients with early-stage cervical cancer who have strong wish to have a child.


Effects of feeding T-2 toxin on RNA, DNA and protein contents of liver and intestinal mucosa of rats.
Young male albino rats were fed 1.5 mg T-2 toxin/kg body weight daily for 4 days and the effects on body weight, liver weight and protein, RNA, and DNA contents of liver and intestinal mucosa were studied. A significant decrease in body weight and an increase in liver weight were observed. Liver protein and DNA and intestinal mucosal protein contents were significantly decreased, whereas intestinal mucosal RNA content was increased. Decreased liver and intestinal mucosal protein synthesis in T-2 toxin-fed rats was inferred from the [14C]leucine incorporation studies.


Electrophysiological evidence of crossmodal correspondence between auditory pitch and visual elevation affecting inhibition of return.
Inhibition of return (IOR) has proved to be weakened by audiovisual integration because of the increased perceptual salience of targets. Although other audiovisual interactions, such as crossmodal correspondence, have also been shown to facilitate attentional processes, to the best of our knowledge, no study has investigated the interaction between crossmodal correspondence and IOR. The present study employed Posner's spatial cueing paradigm and manipulated the cue validity, crossmodal correspondence congruency and time interval of auditory and visual stimuli (AV interval) to explore the effect of crossmodal correspondence on the IOR effect. The behavioral results showed a reduced IOR effect under the correspondence congruency condition in contrast to the correspondence incongruency condition at the AV interval of 200 ms, whereas at an AV interval of 80 ms, the decreased IOR effect under crossmodal correspondence congruency was eliminated. The electrophysiological results showed a reduced amplitude difference in P2 between valid and invalid cue conditions when the crossmodal correspondence effect decreased the IOR effect. The present study provided the first evidence of the weakened effect of the crossmodal correspondence effect on the IOR effect, which could be eliminated by audiovisual integration.


Seizure reduction in TSC2-mutant mouse model by an mTOR catalytic inhibitor.
Tuberous sclerosis complex (TSC) is a neurodevelopmental disorder caused by autosomal-dominant pathogenic variants in either the TSC1 or TSC2 gene, and it is characterized by hamartomas in multiple organs, such as skin, kidney, lung, and brain. These changes can result in epilepsy, learning disabilities, and behavioral complications, among others. The mechanistic link between TSC and the mechanistic target of the rapamycin (mTOR) pathway is well established, thus mTOR inhibitors can potentially be used to treat the clinical manifestations of the disorder, including epilepsy. In this study, we tested the efficacy of a novel mTOR catalytic inhibitor (here named Tool Compound 1 or TC1) previously reported to be more brain-penetrant compared with other mTOR inhibitors. Using a well-characterized hypomorphic Tsc2 mouse model, which displays a translationally relevant seizure phenotype, we tested the efficacy of TC1. Our results show that chronic treatment with this novel mTOR catalytic inhibitor (TC1), which affects both the mTORC1 and mTORC2 signaling complexes, reduces seizure burden, and extends the survival of Tsc2 hypomorphic mice, restoring species typical weight gain over development. Novel mTOR catalytic inhibitor TC1 exhibits a promising therapeutic option in the treatment of TSC.


Role of oxidative DNA damage in mitochondrial dysfunction and Huntington's disease pathogenesis.
Huntington's disease (HD) is a neurodegenerative disorder with an autosomal dominant expression pattern and typically a late-onset appearance. HD is a movement disorder with a heterogeneous phenotype characterized by involuntary dance-like gait, bioenergetic deficits, motor impairment, and cognitive and psychiatric deficits. Compelling evidence suggests that increased oxidative stress and mitochondrial dysfunction may underlie HD pathogenesis. However, the exact mechanisms underlying mutant huntingtin-induced neurological toxicity remain unclear. The objective of this paper is to review recent literature regarding the role of oxidative DNA damage in mitochondrial dysfunction and HD pathogenesis.


Behavioral science at the crossroads in public health: extending horizons, envisioning the future.
The social and behavioral sciences are at a crossroads in public health. In this paper, we attempt to describe a path toward the further integration of the natural and behavioral sciences with respect to the study of behavior and health. Three innovations are proposed. First, we extend and modify the "stream of causation" metaphor along two axes: time, and levels of nested systems of social and biological organization. Second, we address the question of whether 'upstream' features of social context are causes of disease, fundamental or otherwise. Finally, we propose the concept of a risk regulator to advance the study of behavior and health in populations. To illustrate the potential of these innovations, we develop a multilevel framework for the study of health behaviors and obesity in social and biological context.


Estradiol attenuates tau hyperphosphorylation induced by upregulation of protein kinase-A.
Protein kinase A (PKA) plays a crucial role in tau hyperphosphorylation, an early event of Alzheimer disease (AD), and 17beta-estradiol replacement in aging women forestalls the onset of AD. However, the role of estradiol in PKA-induced tau hyperphosphorylation is not known. Here, we investigated the effect of 17beta-estradiol on cAMP/PKA activity and the PKA-induced tau hyperphosphorylation in HEK293 cells stably expressing tau441. We found that 17beta-estradiol effectively attenuated forskolin-induced overactivation of PKA and elevation of cAMP, and thus prevented tau from hyperphosphorylation. These data provide the first evidence that 17beta-estradiol can inhibit PKA overactivation and the PKA-induced tau hyperphosphorylation, implying a preventive role of 17beta-estradiol in AD-like tau pathology.


Effects of increasing age, dosage, and duration of PTH treatment on BMD increase--a meta-analysis.
We studied the effects of increasing age, dosage, and duration of parathyroid hormone (PTH) treatment on changes in bone mineral density (BMD). Randomized placebo controlled trials on PTH treatment in men or women were retrieved from PubMed (1951 to present), Web of Science (1945 to present), or Embase (1974 to present). The search date was November 16, 2010. All studies comparing PTH treatment to either placebo or antiresorptive drugs--for example, bisphosphonates or hormone replacement therapy--were included. A total of 214 studies were identified in the initial search, and 15 of these trials were included. By metaregression analysis, we found that the increase in spine BMD (Z-score) after PTH treatment was blunted by increasing age (R(2) = 0.27; 2p = 0.01, slope -0.023 Z-scores per year, 11 studies). By increasing PTH dosage (μg/d), spine BMD increased significantly (2p = 0.002) with a slope of +0.011 Z-scores/μg/d of teriparatide. Furthermore, the duration of treatment was positively correlated to spine BMD (P < 0.001) with a slope of +0.043 Z-score for each extra month of treatment. We evaluated the BMD effect in hips and found no age dependency (R(2) = 0.04; P = 0.66; 8 studies). However, for the spine, we found a significant relation to daily dosage (P = 0.011), Z-score coefficient 0.0051 ± 0.0020 (2p < 0.01). The treatment duration also correlated positively by a Z-score coefficient of 0.0170 ± 0.0053, 2p < 0.01 per extra month of treatment. PTH treatment alone seems to be able to improve BMD significantly. However, the BMD increase was significantly lower with increasing age in the spine. No age dependency was observed in the hips. In general the effect of treatment was improved with increasing dosage and duration of treatment from 6 to 36 months.


Event segmentation protects emotional memories from competing experiences encoded close in time.
Fear memories are characterized by their permanence and a fierce resistance to unlearning by new experiences. We considered whether this durability involves a process of memory segmentation that separates competing experiences. To address this question, we used an emotional learning task designed to measure recognition memory for category exemplars encoded during competing experiences of fear-conditioning and extinction. Here we show that people recognized more fear-conditioned exemplars encoded during conditioning than conceptually related exemplars encoded immediately after a perceptual event boundary separating conditioning from extinction. Selective episodic memory depended on a period of consolidation, an explicit break between competing experiences, and was unrelated to within-session arousal or the explicit realization of a transition from conditioning to extinction. Collectively, these findings suggest that event boundaries guide selective consolidation to prioritize emotional information in memory-at the expense of related but conflicting information experienced shortly thereafter. We put forward a model whereby event boundaries bifurcate related memory traces for incompatible experiences. This stands in contrast to a mechanism that integrates related experiences for adaptive generalization123, and reveals a potentially distinct organization by which competing memories are adaptively segmented to select and protect nascent fear memories from immediate sources of interference.


Evaluating Changes to Blood-Brain Barrier Integrity in Brain Metastasis over Time and after Radiation Treatment.
The incidence of brain metastasis due to breast cancer is increasing, and prognosis is poor. Treatment is challenging because the blood-brain barrier (BBB) limits efficacy of systemic therapies. In this work, we develop a clinically relevant whole brain radiotherapy (WBRT) plan to investigate the impact of radiation on brain metastasis development and BBB permeability in a murine model. We hypothesize that radiotherapy will decrease tumor burden and increase tumor permeability, which could offer a mechanism to increase drug uptake in brain metastases. Contrast-enhanced magnetic resonance imaging (MRI) and high-resolution anatomical MRI were used to evaluate BBB integrity associated with brain metastases due to breast cancer in the MDA-MB-231-BR-HER2 model during their natural development. Novel image-guided microirradiation technology was employed to develop WBRT treatment plans and to investigate if this altered brain metastatic growth or permeability. Histology and immunohistochemistry were performed on whole brain slices corresponding with MRI to validate and further investigate radiological findings. Herein, we show successful implementation of microirradiation technology that can deliver WBRT to small animals. We further report that WBRT following diagnosis of brain metastasis can mitigate, but not eliminate, tumor growth in the MDA-MB-231-BR-HER2 model. Moreover, radiotherapy did not impact BBB permeability associated with metastases. Clinically relevant WBRT is not curative when delivered after MRI-detectable tumors have developed in this model. A dose of 20 Gy in 2 fractions was not sufficient to increase tumor permeability such that it could be used as a method to increase systemic drug uptake in brain metastasis.


Molecular mechanisms for discrete nitric oxide levels in cancer.
Nitric oxide (NO) has been invoked in nearly every normal and pathological condition associated with human physiology. In tumor biology, nitrogen oxides have both positive and negative affects as they have been implicated in both promoting and preventing cancer. Our work has focused on NO chemistry and how it correlates with cytotoxicity and cancer. Toward this end, we have studied both concentration- and time-dependent NO regulation of specific signaling pathways in response to defined nitrosative stress levels that may occur within the tumor microenvironment. Threshold levels of NO required for activation and stabilization of key proteins involved in carcinogenesis including p53, ERK, Akt and HIF have been identified. Importantly, threshold NO levels are further influenced by reactive oxygen species (ROS) including superoxide, which can shift or attenuate NO-mediated signaling as observed in both tumor and endothelial cells. Our studies have been extended to determine levels of NO that are critical during angiogenic response through regulation of the anti-angiogenic agent thrombospondin-1 (TSP-1) and pro-angiogenic agent matrix metalloproteinase-9 (MMP-9). The quantification of redox events at the cellular level has revealed potential mechanisms that may either limit or potentiate tumor growth, and helped define the positive and negative function of nitric oxide in cancer.


Fulfilling prophecy? Sexually transmitted infections and HIV in Indigenous people in Western Australia.
To compare trends and rates of HIV and sexually transmitted infections in Indigenous and non-Indigenous people of Western Australia. Analysis of WA notification data for chlamydia, gonorrhoea, and primary and secondary syphilis in 2002, and for HIV infections from 1983 to 2002. Rates of HIV and sexually transmitted infection by Indigenous status. In 2002, there were 3046 notifications for chlamydia, 1380 for gonorrhoea and 64 for syphilis. When information on Indigenous status was available, Indigenous people accounted for 41% of chlamydia and 76% of gonorrhoea notifications, with Indigenous:non-Indigenous age-standardised rate ratios of 16 (95% CI, 14-17) and 77 (95% CI, 67-88), respectively. Indigenous people accounted for 90.6% of syphilis notifications (age-standardised Indigenous:non-Indigenous rate ratio, 242 [95% CI, 104-561]). From 1985 to 2002, HIV notification rates for non-Indigenous people in WA declined and rates for Indigenous people increased. From 1994 to 2002, there were 421 notifications of HIV infection in WA residents, 52 (12.4%) in Indigenous people and 369 (87.6%) in non-Indigenous people. Indigenous people accounted for 39% and 6.2% of all notifications in WA females and males, respectively. The Indigenous:non-Indigenous rate ratios were 18 (95% CI, 12-29) for females and 2 (95% CI, 1-3) for males. Indigenous Western Australians are at greater risk of HIV transmission than non-Indigenous people. Strategies to prevent further HIV infection in Indigenous Australians should include control of sexually transmitted infections.


Evidence for an evolutionarily conserved interaction between cell wall biosynthesis and flowering in maize and sorghum.
Factors that affect flowering vary among different plant species, and in the grasses in particular the exact mechanism behind this transition is not fully understood. The brown midrib (bm) mutants of maize (Zea mays L.), which have altered cell wall composition, have different flowering dynamics compared to their wild-type counterparts. This is indicative of a link between cell wall biogenesis and flowering. In order to test whether this relationship also exists in other grasses, the flowering dynamics in sorghum (Sorghum bicolor (L.) Moench) were investigated. Sorghum is evolutionarily closely related to maize, and a set of brown midrib (bmr) mutants similar to the maize bm mutants is available, making sorghum a suitable choice for study in this context. We compared the flowering time (time to half-bloom) of several different bmr sorghum lines and their wild-type counterparts. This revealed that the relationship between cell wall composition and flowering was conserved in sorghum. Specifically, the mutant bmr7 flowered significantly earlier than the corresponding wild-type control, whereas the mutants bmr2, bmr4, bmr6, bmr12, and bmr19 flowered later than their wild-type controls. The change in flowering dynamics in several of the brown midrib sorghum lines provides evidence for an evolutionarily conserved mechanism that links cell wall biosynthesis to flowering dynamics. The availability of the sorghum bmr mutants expands the germplasm available to investigate this relationship in further detail.


Inhibition of mevalonate pathway is involved in alendronate-induced cell growth inhibition, but not in cytokine secretion from macrophages in vitro.
Bisphosphonates are antiresorptive drugs used for the treatment of metabolic bone diseases. They can be divided into two different pharmacological classes: nitrogen-containing and non-nitrogen-containing bisphosphonates. Non-nitrogen-containing bisphosphonates, like clodronate, are metabolised to a toxic ATP-analogue preventing osteoclast mediated bone resorption. Nitrogen-containing bisphosphonates, including alendronate, prevent osteoclast function by inhibiting the mevalonate pathway. Clodronate is known to have anti-inflammatory properties while alendronate induces cytokine secretion from lipopolysaccharide- (LPS) induced macrophages. This study investigates whether the cytotoxicity and cytokine production induced by alendronate and LPS could be counteracted by clodronate or products of mevalonate pathway: oxidized low density lipoprotein (ox-LDL), farnesol and geranylgeraniol. Treatment with alendronate increased LPS-induced secretion of IL-1beta, IL-6 and TNF-alpha from RAW 264 macrophages 2.4-, 1.4- and 1.8-fold, respectively. This treatment was cytotoxic for macrophages as indicated by lowered cell viability. Clodronate and ox-LDL both counteracted the cytokine secretion and cytotoxicity of alendronate. Farnesol and geranylgeraniol did neither reverse the cytokine secretion nor reduce the cytotoxicity of alendronate. Clodronate and ox-LDL were able to counteract the effects of alendronate on macrophages in vitro, probably by their known ability to inhibit DNA binding activity of transcription factors, nuclear factor-kappaB (NF-kappaB) and activating protein-1 (AP-1). These findings suggest that inhibition of mevalonate pathway is not the mechanism responsible for the proinflammatory response caused by alendronate, as it is in alendronate-induced apoptosis and prevention of osteoclast function.


MALAT1 functions as a transcriptional promoter of MALAT1::GLI1 fusion for truncated GLI1 protein expression in cancer.
The long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a cancer biomarker. Furthermore, fusion of the MALAT1 gene with glioma-associated oncogene 1 (GLI1) is a diagnostic marker of plexiform fibromyxoma and gastroblastoma; however, the function of this fusion gene remains unexplored. In this study, we elucidate the structure and function of the MALAT1::GLI1 fusion gene. To this end, we determined a transcriptional start site (TSS) and promoter region for truncated GLI1 expression using rapid amplification of the 5' cDNA end and a luciferase reporter assay in cultured cells transfected with a plasmid harboring the MALAT1::GLI1 fusion gene. We found that the TATA box, ETS1 motif, and TSS were located in MALAT1 and that MALAT1 exhibited transcriptional activity and induced expression of GLI1 from the MALAT1::GLI1 fusion gene. Truncated GLI1, lacking SUMOylation and SUFU binding sites and located in the nucleus, upregulated mRNA expression of GLI1 target genes in the hedgehog signaling pathway. We demonstrate a distinct and alternative function of MALAT1 as a transcriptional promoter for expression of the MALAT1::GLI1 fusion gene. Our findings will aid future research on MALAT1 and its fusion gene partners.


Utilization of concentrate of membrane filtration of bleach plant effluent in brick production.
Utilization potential of membrane filtration retentate (concentrate) of bleach plant effluent from paper industry, in bricks production, was investigated in the present study. Bricks were prepared by using retentate of membrane filtration in place of water. The physical properties measured for bricks were dimensions, density, moisture content, compressive strength, water absorption and porosity. Chemical analysis of the bricks was performed for water leachability and acid leachability, using ASTM Standard methods D 3987-06 and D 5233-92 respectively. The leachate samples were analyzed for Al, Cu, Fe, Co, Cr, Pb, Ni and Zn using AAS (atomic absorption spectrophotometer). Adsorbable organic halides (AOX) of leachate were analyzed by using AOX analyzer model ECS 1200 using column method. It was observed that the physical and chemical properties of bricks especially fire clay bricks were not adversely affected by the use of wastewater. Therefore, the organic matter of the retentate of membrane filtration creates no problems for this application.


Quantifying the surface characteristics and flocculability of Ralstonia eutropha.
The microbial surface and flocculability were qualitatively characterized through the combination of the surface thermodynamic and the extended DLVO approaches, with Ralstonia eutropha, a polyhydroxybutyrate-producing bacterium, as an example. The negativity of the zeta potential of R. eutropha decreased from the initial -19.5 to -11 mV in its cultivation with the consumption of glucose. The total interfacial free energy (DeltaGadh) was changed from -80 to 28.5 mJ m(-2) in its entire growth process. This suggests that the bacterial surface changed from hydrophobic into hydrophilic, resulting in an alteration of its surface characteristics and flocculability in its different growth phases. As a result, the stability ratio of suspensions increased with the increasing cultivation time, indicating that the cell particles became more repulsive with each other and led to a more stable suspension of R. eutropha in its cultivation. The obtained information in this work might be useful for better understanding the surface characteristics and the flocculability and even manipulating its flocculability in the microbial growth process.


Tobacco Smoke Exposure, School Engagement, School Success, and Afterschool Activity Participation Among US Children.
Tobacco smoke exposure (TSE), defined as secondhand smoke (SHS) and thirdhand smoke (THS), is associated with negative health consequences. This study's objective was to assess the associations between home TSE status and school engagement, school success, and afterschool activity participation among school-aged children. We conducted a secondary analysis of 2018-2019 National Survey of Children's Health cross-sectional data. Children ages 6-11 years (N = 17,466) were categorized into home TSE groups: no home TSE; THS exposure only; and SHS and THS exposure. Weighted logistic and Poisson regression models were built. Compared to children with no home TSE, children with home THS exposure only and SHS and THS exposure were at decreased odds of being engaged in school (AOR = 0.69, 95%CI = 0.57, 0.83; AOR = 0.63, 95%CI = 0.41, 0.97, respectively), and at increased odds of having ≥1 school-to-home contact about child problems in school (AOR = 1.83, 95%CI = 1.50, 2.23; AOR = 1.58, 95%CI = 1.05, 2.37, respectively). Children with THS exposure only were at increased odds of missing ≥1 school day (AOR = 1.43, 95%CI = 1.13, 1.81). Children with THS exposure only (ARR = 0.90, 95%CI = 0.83, 0.96) and SHS and THS exposure (ARR = 0.74, 95%CI = 0.61, 0.89) were at reduced likelihood of participating in a higher number of afterschool activities. Children exposed to home tobacco smoke are at unique risk for poorer school engagement and success.


Absorption and excretion of 3H-raubasine in human subjects and dogs.
The pharmacokinetic behavior of (3,5,6-3H)-raubasine (RAU) was investigated in human subjects after oral administration and in dogs after both intravenous and oral administration. By the oral route RAU peak plasma levels appeared in human subjects after 1 h and in dogs after 2 h. Three-day cumulative urinary excretion was 22% by i.v. route and 13% by oral route in dogs, being 29% in human subjects after oral administration. Three-day cumulative faecal excretion in dogs was 51% by i.v. route and 57% by oral route whilst it was 24% in humans. From a comparison between the urinary excretion values observed after i.v. and oral administration, a RAU intestinal absorption of 59% may be obtained.


Screening of Korean medicinal plants for lipase inhibitory activity.
The pancreatic lipase inhibitory activity of the aqueous ethanol extracts obtained from 19 medicinal plants was evaluated in vitro by a continuous-monitoring pH-Stat technique using tributyrin as a substrate. Of the extracts tested, those of Juniperus communis (bark) and Illicium religiosum (wood) exhibited the strongest activity with an IC50 value of 20.4 and 21.9 microg/mL, respectively.


Pitavastatin regulates helper T-cell differentiation and ameliorates autoimmune myocarditis in mice.
Experimental autoimmune myocarditis (EAM) is a mouse model of inflammatory cardiomyopathy, and the involvement of T helper (Th) 1 and Th17 cytokines has been demonstrated. Accumulated evidence has shown that statins have anti-inflammatory and immunomodulatory effects; however, the mechanism has not been fully elucidated. This study was designed to test the hypothesis that pitavastatin affects T cell-mediated autoimmunity through inhibiting Th1 and Th17 responses and reduces the severity of EAM in mice. The EAM model was established in BALB/c mice by immunization with murine α-myosin heavy chain. Mice were fed pitavastatin (5 mg/kg) or vehicle once daily for 3 weeks from day 0 to day 21 after immunization. Pitavastatin reduced the pathophysiological severity of the myocarditis. Pitavastatin treatment inhibited the phosphorylation of signal transducer and activator of transcription (STAT)3 and STAT4, which have key roles in the Th1 and Th17 lineage commitment, respectively, in the heart, and suppressed production of Th1 cytokine interferon-γ and Th17 cytokine interleukin-17 from autoreactive CD4(+) T cells. In in vitro T-cell differentiation experiments, pitavastatin-treated T cells failed to differentiate into Th1 and Th17 cells through inhibiting the transcription of T-box expressed in T-cells (T-bet) and RAR-related orphan receptor γt (RORγT) which have critical roles in the development of Th1 and Th17 cells, respectively, and this failure was rescued by adding mevalonate. Pitavastatin inhibits Th1 and Th17 responses and ameliorates EAM. These results suggest that statins may be a promising novel therapeutic strategy for the clinical treatment of myocarditis and inflammatory cardiomyopathy.


Expression profile of SARS-CoV-2 cellular entry proteins in normal oral mucosa and oral squamous cell carcinoma.
Besides angiotensin converting enzyme 2 (ACE2), an active involvement of proteases (FURIN and/or TMPRSS2) is important for cellular entry of SARS-CoV-2. Therefore, a simultaneous expression profiling of entry proteins in a tissue might provide a better risk assessment of SARS-CoV-2 infection as compared to individual proteins. In an attempt to understand the relative susceptibility of oral squamous cell carcinoma (OSCC) lesions as compared to the normal oral mucosa (NOM) for SARS-CoV-2 attachment/entry, this study examined the mRNA and protein expression profiles of ACE2, FURIN, and TMPRSS2 in the corresponding tissues using public transcriptomic and proteomics datasets. Public transcriptomic and proteomics datasets (the Cancer Genome Atlas (TCGA)/the Genotype-Tissue Expression (GTEx), the Human Protein Atlas (HPA), and two independent microarray datasets) were used to examine the expression profiles of ACE2, TMPRSS2 and FURIN in NOM and OSCC. ACE2, TMPRSS2, and FURIN mRNAs were detected in NOM, however, at lower levels as compared to other body tissues. Except for moderate up-regulation of FURIN, expression levels of ACE2 and TMPRSS2 mRNA were unchanged/down-regulated in OSCC as compared to the NOM. These results indicate that NOM may serve as a possible site for SARS-CoV-2 attachment, however, to a lesser extent as compared to organs with higher expression levels of the SARS-CoV-2 entry proteins. However, the evidence is lacking to suggest that expression status of entry proteins predisposes OSCC lesions to additional risk for SARS-CoV-2 attachment/entry as compared to NOM.


Individual and environmental contingencies associated with multiple suicide attempts among U.S. military personnel.
Suicidal behavior among U.S. military personnel persists as a significant public health issue. Previous research indicates the primary motive for suicide attempts among military personnel is the desire to reduce or alleviate emotional distress, a finding that converges with studies in nonmilitary samples. Much less is understood about the consequences of a first suicide attempt that could influence the occurrence of additional suicide attempts. In order to identify these contingencies, 134 active duty Soldiers who had attempted suicide (n=69 first-time attempters, n=65 multiple attempters) participated in structured interviews focused on their experiences immediately following their first attempt. Soldiers were more likely to have made multiple suicide attempts if they were younger at the time of their first attempt, were not admitted to a hospital or treatment program after their first attempt, or experienced emotional and psychological relief immediately afterwards. Results suggest that Soldiers who experience emotional and/or psychological relief immediately after their first suicide attempt or do not receive treatment are more likely to make additional suicide attempts.


Impact of physical fitness on salivary stress markers in sedentary to low-active young to middle-aged men.
Salivary cortisol (sCort) and alpha-amylase (sAA) are popular measurement tools for investigating hypothalamic-pituitary-adrenal and autonomic functioning, respectively. Despite their widespread use, only little is known about whether lifestyle factors such as regular exercise and physical fitness impact these two salivary stress markers. The lack of an objectively assessed fitness status is a major drawback of nearly all previous studies in this area. Our main objective was therefore to examine the impact of fitness status on sCort and sAA responses to a validated stressor. Sedentary to low-active men (N=115, 45.7±10.1 years, 26.7±3.4kg/m(2)) were exposed to the Trier Social Stress Test for Groups and provided saliva samples before and repeatedly after the stressor. An ergometer lactate exercise test provided information on physical fitness, that is, power achieved at the individual anaerobic threshold relative to body weight. From age-specific percentiles, a 2- (n=65 low fitness, n=50 high fitness) and a 3-group (n=37 low fitness, n=49 moderate fitness, n=29 high fitness) solution were calculated to elucidate the impact of different fitness levels. Prior to stress testing, sCort and sAA levels were similar in all groups. While both markers increased significantly due to stress exposure, fitness effects appeared rather small. When applying the 3-group solution, we observed attenuated sCort slopes in high- and moderate-fit men. Although sAA response slopes were pronounced in high-fit men, they failed to reach statistical significance. Mean stress reactivity (area under the curve with respect to ground, baseline, and response maximum) did not differ between groups and no association with fitness level was found. Our findings suggest only a weak effect of objectively assessed physical fitness on biological stress responses with attenuated hypothalamic-pituitary-adrenal but (non-significantly) higher autonomic responses in fitter men. Physical fitness is a factor that should not be neglected in future studies employing salivary stress markers, especially in studies interested in acute stress responses.


The anterior basal lobe and control of prey-capture in the cuttlefish (Sepia officinalis).
The predatory behaviour of the cuttlefish comprises several stages: prey-detection, orientation, translation and prey-seizing. In this neuroethological study, lesions to the anterior basal lobe were made by an electrolytic method and the animals were allowed to attack their prey in an unrestricted way so that motor defects, functional recovery and the emergence of new adaptative behavioural strategies could be studied. Lesions to the central region of the anterior basal lobe suppress the orientating behaviour, thus only prey situated in the frontal visual field can be seized. Less extensive lesions in this region are associated with similar defects. Without head orientation, the cuttlefish still rotates with its fins. This rotation, however, is usually underestimated, tentacular ejection thus missing the prey. Dorsal lesions cause an underestimation of tentacular strike often associated with defects in maintaining ocular convergence. These results demonstrate the heterogenous function of the anterior basal lobe and its complex role in the control of predatory behaviour.


A technique for simulating visual field losses in virtual environments to study human navigation.
The following paper describes a new technique for simulating peripheral field losses in virtual environments to study the roles of the central and peripheral visual fields during navigation. Based on Geisler and Perry's (2002) gaze-contingent multiresolution display concept, the technique extends their methodology to work with three-dimensional images that are both transformed and rendered in real time by a computer graphics system. In order to assess the usefulness of this method for studying visual field losses, an experiment was run in which seven participants were required to walk to a target tree in a virtual forest as quickly and efficiently as possible while artificial head and eye-based delays were systematically introduced. Bilinear fits were applied to the mean trial times in order to assess at what delay lengths breaks in performance could be observed. Results suggest that breaks occur beyond the current delays inherent in the system. Increases in trial times across all delays tested were also observed when simulated peripheral field losses were applied compared to full FOV conditions. Possible applications and limitations of the system are discussed. The source code needed to program visual field losses can be found at lions.med.jhu.edu/archive/turanolab/Simulated_Visual_Field_Loss_Code.html.


Genotoxicity, metabolism and blood kinetics of epichlorohydrin in mice.
Epichlorohydrin (ECH), a direct mutagen in vitro, did not induce chromosomal aberrations in bone-marrow cells of CD1 mice given single oral doses of 50 and 200 mg/kg in water. The ECH diol derivative (3-chloro-1,2-propanediol) was tested in vitro by a forward-mutation assay on the yeast Schizosaccharomyces pombe and showed a weak but significant mutagenic effect. The failure of ECH to induce mutagenic effects appears to be due to the rapid metabolic clearance of the compound in vivo. ECH blood kinetics at both doses, and at the same time the concentration of the diol, were determined. ECH rapidly disappeared from mouse blood, being no longer detectable 20 min after treatment. In contrast, 3-chloro-1,2-propanediol was measurable up to 5 h after dosage. No difference was observed in the kinetic and metabolic behavior of ECH after single and repeated doses (50 and 200 mg/kg/day for 7 days). When 3-chloro-1,2-propanediol was tested, neither glutathione depletion nor epoxide hydrolase inhibition (evaluated with both styrene-7,8-oxide and ECH as substrates) could be detected in mouse liver. Finally, no difference in ECH blood kinetics or metabolism were observed in experiments in which the compound was administered (200 mg/kg) intraperitoneally in water or orally as a solution in dimethyl sulfoxide.


The effect of sepsis and septic shock on the viscoelastic properties of clot quality and mass using rotational thromboelastometry: A prospective observational study.
The study purpose was to define changes in coagulation across the sepsis spectrum using rotational thromboelastometry (ROTEM). Sepsis patients were recruited on admission to the Emergency Department and Intensive Care Units of a large teaching hospital in Wales. ROTEM markers of clot development and fibrinolysis were determined, as well as standard coagulation markers. A healthy control group matched for age and gender was also recruited (n=44). 100 patients were recruited (50 sepsis, 20 severe sepsis and 30 septic shock). Maximum clot firmness was significantly higher in the sepsis (p<0.001) and severe sepsis (p=0.012) groups than the healthy control (71.6±4.5 and 70.4±4.1 vs 64.4 respectively). In septic shock there was prolonged clot development; however, maximum clot firmness remained normal. Fibrinolytic function was significantly impaired in septic shock, which was also significantly associated with 28-day mortality (p<0.001). ROTEM indicated significantly enhanced clot structural development in sepsis and severe sepsis, which could be indicative of a hypercoagulable phase. In septic shock, despite there being a prolongation of clotting pathways and impaired fibrinolysis, clot mass was comparably normal, suggestive of the development of a clot with healthy characteristics.


Micromechanical studies of mitotic chromosomes.
Mitotic chromosomes respond elastically to forces in the nanonewton range, a property important to transduction of stresses used as mechanical regulatory signals during cell division. In addition to being important biologically, chromosome elasticity can be used as a tool for investigating the folding of chromatin. This paper reviews experiments studying stretching and bending stiffness of mitotic chromosomes, plus experiments where changes in chromosome elasticity resulting from chemical and enzyme treatments were used to analyse connectivity of chromatin inside chromosomes. Experiments with nucleases indicate that non-DNA elements constraining mitotic chromatin must be isolated from one another, leading to the conclusion that mitotic chromosomes have a chromatin 'network' or 'gel' organization, with stretches of chromatin strung between 'crosslinking' points. The as-yet unresolved questions of the identities of the putative chromatin crosslinkers and their organization inside mitotic chromosomes are discussed.


Nucleotide sequence and structure of the Klebsiella pneumoniae pqq operon.
A 6940 bp Klebsiella pneumoniae chromosomal DNA fragment, containing genes involved in pyrroloquinoline quinone (PQQ) biosynthesis, was sequenced. Six open reading frames, pqqA, pqqB, pqqC, pqqD, pqqE and pqqF were identified in the pqq operon, which coded for polypeptides of 2764 (23 amino acids), 33,464, 28,986, 10,436, 42,881 and 83,616 Da, respectively. The transcription startpoint was mapped by primer extension analysis, upstream of pqqA, and promoter boxes could be identified. The gene products of pqqB, pqqC, pqqE and pqqF were detected in maxi-cells and the molecular weights of the proteins corresponded with the molecular weights deduced from the nucleotide sequence. The gene products of pqqA, pqqB, pqqC, pqqD and pqqE show 49%-64% identity in amino acid sequence with those of pqqIV, pqqV, pqqI, pqqII and pqqIII respectively in the cloned pqq cluster of Acinetobacter calcoaceticus. The 84 kDa protein encoded by pqqF, which is not present in the cloned pqq cluster of A. calcoaceticus but which is essential for PQQ biosynthesis in K. pneumoniae and Escherichia coli, seems to belong to a family of proteases.


Effects of prolonged working hours on heart rate variability in internal medicine physicians.
Prior studies have utilized heart rate variability (HRV) as the assessment tools for psychological and physiological stress during 24-h shift. However, data regarding effects of prolonged working hours > 24 h on HRV are limited. We aimed to compare between pre- and post-call HRV among physicians who worked 24 plus 8 h. The study included 60 physicians in the internal medicine training. All subjects underwent Holter ECG monitoring for HRV assessment. We compared between HRV of an 8-h regular workday (8am to 4 pm) before on-call duty (pre-call HRV) and an 8-h workday after 24-h on-call duty (post-call HRV). The mean age was 26 ± 2.5 years. Mean total sleep time during on-call duty was 238.9 ± 88.3 min. In overall population, the time-domain and frequency-domain HRV parameters were not different between pre- and post-call day. However, the physicians reported their sleep time in the 1st quartile (< 180 min) had significant increase in SDNN, pNN50, high frequency (HF), and decrease in low/high frequency ratio (LF/HF). In contrast, the physicians reported their sleep time in the 4th quartile (> 307.5 min) had significant decrease in pNN50, LF, HF, and increase in heart rate. Multiple linear regression revealed total sleep time as an independent factor associated with pre- and post-call HRV alterations. More sleep during on call (> 5 h) was associated with HRV pattern suggesting both increased sympathetic activity and reduced parasympathetic activity, while less sleep (< 3 h) during on call was associated with post-call parasympathetic rebound HRV pattern.


Congenital flexion deformity of the long, ring, and little fingers with an aberrant origin of the flexor digitorum profundus: case report.
A case of congenital flexion deformity of the long, ring, and little fingers with an aberrant origin of the flexor digitorum profundus is described. The aberrant tendinous band originated from a bone prominence in the medial and anterior aspect of the proximal side of the ulna, which was confirmed in the operation. Resection of the aberrant origin could achieve only partial relief for the flexion contracture of the 3 fingers. Further muscle-sliding procedure was needed to achieve a thorough release.


Development of Polymeric Micelles of Oleanolic Acid and Evaluation of Their Clinical Efficacy.
Oleanolic acid has been used only as a subsidiary agent in cosmetic products. The aim of the study is to show the effect of oleanolic acid as an active ingredient for the alleviation of wrinkles in humans and to develop a polymeric micelle formulation that enables poorly soluble oleanolic acid to be used as a main ingredient in cosmetic products for reducing wrinkles. The solubility of oleanolic acid was evaluated in solubilizers, surfactants, and polymers. The particle sizes and shapes of polymeric micelles containing oleanolic acid were evaluated by electrophoretic light scattering spectrophotometer and scanning electron cryomicroscopy. Encapsulation efficiency and skin permeation were measured by HPLC. Stability of the polymeric micelles stored at 40 °C for 3 months was evaluated by visual observation, particle size measurement, and oleanolic acid content measurement. Polymeric micelles in final product ampoule form were applied around the eyes of 23 female subjects for 8 weeks. Five skin parameters were evaluated by optical profilometry every 4 weeks for 8 weeks. In addition, professionals made visual observations of the skin and a human skin irritation study was conducted. Polymeric micelles of oleanolic acid with a particle size of less than 100 nm were prepared using Capryol 90® and poloxamer. The skin permeation rate of the oleanolic acid in the polymeric micelles was higher than that in the other solutions made of oleanolic acid dispersed in 2 different surfactants. No significant changes in particle size, color, or oleanolic acid content were observed, and the polymeric micelles stored at 40 °C for 3 months did not undergo phase separation. After 8 weeks of application, skin irritation had not developed and all five parameters evaluated by optical profilometry as well as the visual evaluation scores were significantly improved. This study showed that the polymeric micelles of oleanolic acid prepared in this study were stable and effective at alleviating wrinkles in humans as the principal active ingredient. Based on these findings, it is expected that polymeric micelles of oleanolic acid can be widely used in cosmetic applications.


Triolein breath test: a sensitive and specific test for fat malabsorption.
In order to develop a simple and accurate screening test for steatorrhea, the authors compared quantitative stool fat excretion with breath excretion of 14CO2 after the ingestion of 14C-labeled trioctanoin, tripalmitin, or triolein. The study group included 24 nonobese subjects without steatorrhea but with diarrhea secondary to irritable bowel syndrome and 40 subjects with steatorrhea. The triolein breath test provided the most reliable discrimination, with 100% sensitivity and 96% specificity. The trioctanoin and tripalmitin breath tests were sensitive, but lacked specificity (69% and 58% false-positive results, respectively). Three of the 12 obese patients with irritable bowel syndrome had false-positive triolein breath results. In detecting steatorrhea, the triolein breath test was moderately superior to the measurement of serum carotene and to qualitative stool fat. Thus, the triolein breath test appears to be a sensitive, specific, noninvasive, and relatively simple screening test for the detection of steatorrhea.


Involvement of phospholipase D-related signal transduction in chemical-induced programmed cell death in tomato cell cultures.
Phospholipase D (PLD) and its product phosphatidic acid (PA) are incorporated in a complex metabolic network in which the individual PLD isoforms are suggested to regulate specific developmental and stress responses, including plant programmed cell death (PCD). Despite the accumulating knowledge, the mechanisms through which PLD/PA operate during PCD are still poorly understood. In this work, the role of PLDα1 in PCD and the associated caspase-like proteolysis, ethylene and hydrogen peroxide (H(2)O(2)) synthesis in tomato suspension cells was studied. Wild-type (WT) and PLDα1-silenced cell lines were exposed to the cell death-inducing chemicals camptothecin (CPT), fumonisin B1 (FB1) and CdSO(4). A range of caspase inhibitors effectively suppressed CPT-induced PCD in WT cells, but failed to alleviate cell death in PLDα1-deficient cells. Compared to WT, in CPT-treated PLDα1 mutant cells, reduced cell death and decreased production of H(2)O(2) were observed. Application of ethylene significantly enhanced CPT-induced cell death both in WT and PLDα1 mutants. Treatments with the PA derivative lyso-phosphatidic acid and mastoparan (agonist of PLD/PLC signalling downstream of G proteins) caused severe cell death. Inhibitors, specific to PLD and PLC, remarkably decreased the chemical-induced cell death. Taken together with our previous findings, the results suggest that PLDα1 contributes to caspase-like-dependent cell death possibly communicated through PA, reactive oxygen species and ethylene. The dead cells expressed morphological features of PCD such as protoplast shrinkage and nucleus compaction. The presented findings reveal novel elements of PLD/PA-mediated cell death response and suggest that PLDα1 is an important factor in chemical-induced PCD signal transduction.


The interpretation of serial Johne's disease milk antibody results is affected by test characteristics, pattern of test results and parallel bovine tuberculosis testing.
In the UK, quarterly Johne's disease milk antibody ELISAs (JD-mELISAs) are commonly used to classify animals which are likely to be infectious, termed "red cows". "Red cows" are classified following two positive results from the previous four tests (e.g. + - - +). All cattle are also regularly screened for bovine tuberculosis using intradermal avian and bovine tuberculin, and it is advised to maintain a 60 day interval between a tuberculosis test and JD-mELISA. To evaluate the impact of bovine tuberculosis testing on JD-mELISAs, and to quantify the impact of test specificity and "red cow" classification test pattern on the probability of infection. Four years of individual cow milk records with JD-mELISA results were collated from 735 dairy farms and matched to tuberculosis testing records. A two-level multivariable logistic regression model quantified the effect of tuberculosis testing on JD-mELISA result. The specificity and age-dependent sensitivity of a single JD-mELISA were estimated and used to calculate likelihood ratios following each test. Using Bayes' theorem, the posterior probability of infection with Johne's disease was calculated for different specificities, ages of cow, and patterns of test results. There were increased odds of a positive JD-mELISA if it was ≤30 days (OR: 2.1) or 31-60 days (OR: 1.2) after a tuberculosis test, compared to >90 days. A larger avian skin reaction at the tuberculosis test was also associated with increased odds of a positive JD-mELISA. The proportion of cows which tested exclusively negative after their first positive JD-mELISA was higher if that JD-mELISA was ≤30 days after a tuberculosis test compared to >90 days. The posterior probability of infection reduced substantially when the test specificity was slightly reduced. In "red cows" classified following two consecutive positive tests, if the test specificity was reduced to 0.95, then the posterior probability of infection was only >95 % if the prior probability was >13 %. If the "red cow" classification was due to two non-consecutive positive tests (+ - - +), the posterior probability of infection was only >95 % if the prior probability was >43 %. Testing for Johne's disease within 60 days of a tuberculosis test is associated with a higher chance of a positive JD-mELISA and this may reflect a reduction in the ELISA specificity. Relatively small reductions in JD-mELISA specificity can markedly reduce the posterior probability of infection which also depends on the pattern of test results which classifies "red cows".


The oscillating peroxidase-oxidase reaction in an open system. Analysis of the reaction mechanism.
1. The oscillations in the peroxidase-oxidase reaction in an open system with NADH as the hydrogen donor are caused by the reaction starting and stopping at critical concentrations of the substrates O2 and NADH. The existence of such critical concentrations is typical of branched chain reactions. 2. The critical concentrations of O2 and NADH that determine the initiation of the reaction are mutually dependent. 3. The branching reactions that determine these critical concentrations involve compounds I and II. 4. Superoxide may be involved in the branching reactions by reacting with NADH and ferriperoxidase. At pH 5.1 the rate constant for the latter reaction is determined as 1.5 . 10(5) M-1 . s-1, whereas for the former reaction only an upper limit for the rate constant of 3.5 . 10(4) M-1 . s-1 could be estimated. These relatively low rate constants suggest that alternative branching reactions may also be involved.


Analysis of Factors Influencing Insomnia and Construction of a Prediction Model: A Cross-sectional Survey on Rescuers.
To determine the factors influencing insomnia and construct early insomnia warning tools for rescuers to informbest practices for early screening and intervention. Cluster sampling was used to conduct a cross-sectional survey of 1,133 rescuers from one unit in Beijing, China. Logistic regression modeling and R software were used to analyze insomnia-related factors and construct a PRISM model, respectively. The positive rate of insomnia among rescuers was 2.74%. Accounting for participants' age, education, systolic pressure, smoking, per capita family monthly income, psychological resilience, and cognitive emotion regulation, logistic regression analysis revealed that, compared with families with an average monthly income less than 3,000 yuan, the odds ratio ( OR) values and the [95% confidence interval ( CI)] for participants of the following categories were as follows: average monthly family income greater than 5,000 yuan: 2.998 (1.307-6.879), smoking: 4.124 (1.954-8.706), and psychological resilience: 0.960 (0.933-0.988). The ROC curve area of the PRISM model (AUC) = 0.7650, specificity = 0.7169, and sensitivity = 0.7419. Insomnia was related to the participants' per capita family monthly income, smoking habits, and psychological resilience on rescue workers. The PRISM model's good diagnostic value advises its use to screen rescuer early sleep quality. Further, advisable interventions to optimize sleep quality and battle effectiveness include psychological resilience training and smoking cessation.


Comparison of a stable isotope-labeled and an analog internal standard for the quantification of everolimus by a liquid chromatography-tandem mass spectrometry method.
Everolimus is an immunosuppressant drug used in solid organ transplantation. Immunoassays and liquid chromatography-mass spectrometry (LC-MS) methods have been used for therapeutic drug monitoring of this drug. In LC-tandem mass spectrometry (MS/MS) methods, both 32-desmethoxyrapamycin and everolimus-d4 have been used as internal standards. To compare 2 internal standards (32-desmethoxyrapamycin and everolimus-d4) for the quantification of everolimus by an LC-MS/MS method. Both 32-desmethoxyrapamycin and everolimus-d4 were introduced in the method validation process with 2 transitions simultaneously monitored for everolimus (975.6 → 908.7 as the quantifier and 975.6 → 926.9 as the qualifier) by an established LC-MS/MS method. The key performance characteristics were lower limit of quantification, accuracy, precision, and comparison with an LC-MS/MS method offered by another laboratory. The lower limit of quantification (LLOQ) was 1.0 ng/mL using either internal standard with an analytical recovery of 98.3%-108.1% across the linear range. The total coefficient of variation for everolimus was 4.3%-7.2% with no significant difference between the 2 internal standards. In comparison with an independent LC-MS/MS method, though everolimus-d4 offered a better slope (0.95 versus 0.83), both internal standards showed acceptable results and had a coefficient of correlation r > 0.98 in the tested concentration range of 1.2-12.7 ng/mL. Although everolimus-d4 offered a more favorable comparison with an independent LC-MS/MS method, both everolimus-d4 and 32-desmethoxyrapamycin had acceptable performance as the internal standards for everolimus quantification by the LC-MS/MS method.