Purification and properties of hyaluronidase from bull sperm.
Hyaluronidase from bull sperm was fractionated by ammonium sulfate and further purified by DEAE-cellulose and Sephadex chromatography. The highly purified hyaluronidase preparation showed 2,370 units per mg of protein (68,730 N.F. units per mg of protein), i.e. 182-fold purification. Disc gel electrophoresis showed one major component. The molecular weight of bull sperm hyaluronidase was 62,000 by sodium dodecyl sulfate gel electrophoresis. Hyaluronidase from bull sperm has optimum activity at pH 3.8 and an absolute requirement for cations. Kplus and Naplus have a greater effect than Ca2plus, Mg2plus, and Mn2plus, whereas Co2plus, Cu2plus, and Zn2plus do not affect the enzyme activity. Purified preparations are less stable than crude extracts stored frozen at minus 15 degrees. Km of hyaluronidase with hyaluronic acid as substrate is 3.7 mg per ml and Vmax is 2.4 mumol per min by Hofstee plot.


Statistical inferences of extended concentration indices for directly standardized rates.
The relative concentration index (RCI) and the absolute concentration index (ACI) have been widely used for monitoring health disparities with ranked health determinants. The RCI has been extended to allow value judgments about inequality aversion by Pereira in 1998 and by Wagstaff in 2002. Previous studies of the extended RCI have focused on survey sample data. This paper adapts the extended RCI for use with directly standardized rates (DSRs) calculated from population-based surveillance data. A Taylor series linearization (TL)-based variance estimator is developed and evaluated using simulations. A simulation-based Monte Carlo (MC) variance estimator is also evaluated as a comparison. Following Wagstaff's approach in 1991, we extend the ACI for use with DSRs. In all simulations, both the TL and MC methods produce valid variance estimates. The TL variance estimator has a simple, closed form that is attractive to users without sophisticated programming skills. The TL and MC estimators have been incorporated into a beta version of the National Cancer Institute's Health Disparities Calculator, a free statistical software tool that enables the estimation of 11 commonly used summary measures of health disparities for DSRs.


Age-related treatment patterns in sickle cell disease patients and the associated sickle cell complications and healthcare costs.
This study explored the blood transfusion patterns, SCD complications, utilization of iron chelation therapies (ICT), healthcare resource use, and costs in pediatric, transitioning (18 years old) and adult patients with SCD. Data from Florida (1998-2009), New Jersey (1996-2009), Missouri (1997-2010), Kansas (2001-2009), and Iowa (1998-2010) state Medicaid were used. Patients with ≥2 SCD diagnoses and ≥1 transfusion event were included. Rates of transfusion events, SCD complications, and proportion of eligible patients receiving ICT were calculated. ICT eligibility was defined as receiving ≥10 transfusions over lifetime. SCD complications included pain, pulmonary event, infection event, renal, cardiovascular, stroke, leg ulcers, and avascular necrosis. Regressions were used to assess risk factors for transfusion and identify the main drivers of costs. The sample included 3,208 patients. The transfusion rate increased from 1-year-old to a peak at 16 years old, then dropped until age 26 and remained stable thereafter. In contrast the frequency of diagnoses for SCD complications increased markedly after age 16. Post-transition patients (≥18 years old) were significantly associated with fewer transfusions (odds ratio: 0.80, P = 0.002). Among eligible patients for ICT, there was no statistically significant difference in total cost between the ICT and no ICT groups (adjusted cost difference, $136, P = 0.114). Patients transitioning to adult care received less transfusions and hydroxyurea, less ICT when eligible for chelation therapy, had higher healthcare costs and suffered from more frequent SCD related complications than pediatric patients. These findings highlight the changes in treatment patterns corresponding to transition to adult care.


Investigating morphological changes in treated vs. untreated stone building materials by x-ray micro-CT.
Calcareous stones have been largely used to build historical buildings. Among these, the calcarenites are usually characterized by a high content of calcite and a high open porosity, which make them very sensitive to the weathering caused by physical and chemical agents. In order to prevent their deterioration and to retard their decay, different protective products-mainly polymers-are applied on the stone artefact surfaces. In this work we apply the methodology tested in a preliminary study to investigate the morphological changes of the internal structure of a biocalcarenite (Lecce stone) by micro x-ray computed tomography (micro-CT). The porosity and other morphological parameters of the rock before and after the conservation treatment were calculated on a significant number of samples. The Student's t test was applied for statistical comparison. The results reveal that the treatment with Paraloid B72 (PB 72) is homogenously distributed and causes small changes to the natural properties of the rock, whereas the application of a fluoroelastomer (NH) causes an appreciable decrease in porosity and variation in terms of wall thickness distribution, probably resulting from its inhomogeneous distribution.


Tracing the sources of nitrate in the Han River watershed in Korea, using delta15N-NO3- and delta18O-NO3- values.
The dissolved nitrate concentrations and their nitrogen and oxygen isotopic ratios were analyzed in seasonal samples from Korea's Han River to ascertain the seasonal and spatial variations of dissolved nitrate and its possible sources. Nitrate concentrations in the South Han River (SHR) were much higher than those in the North Han River (NHR), probably because of the more extensive distribution of agricultural fields, residential areas and animal farms in the SHR drainage basin. The nitrogen isotopic composition of dissolved nitrate indicates that nitrate-nitrogen (NO(3)(-)-N) is derived mainly from atmospheric deposition and/or soil organic matter in the NHR but comes principally from manure or sewage, with only a minor contribution from atmospheric deposition or soil organic matter, in the SHR. The oxygen isotopic compositions of dissolved nitrate suggest that most atmospheric nitrate undergoes microbial nitrification before entering the river.


Scope and perspectives of intravital microscopy--bridge over from in vitro to in vivo.
As observations in vitro become more sophisticated, it is increasingly important to be able to assess their relevance to the intact animal. Intravital microscopy provides an advanced set of tools for the dissection of the in vivo situation.


Strategies to reduce risk factors in hypertensive patients who smoke.
Hypertensive patients who smoke have a high risk of developing premature cardiovascular disease. In addition to encouraging these patients to stop smoking, effective nondrug therapies include weight reduction, salt restriction, and alcohol limitation. For patients whose hypertension is severe or who have other health problems, antihypertensive drug therapy is also used. In the past, diuretics and beta-blockers have proved popular. However, these drugs have produced biochemical disturbances, such as diuretic-induced hypokalemia and both diuretic- and beta-blocker-induced alterations in blood lipids. The hazard of such drug-induced alterations may be greater in hypertensive patients, who may already suffer from hypercholesterolemia. Other drugs are available that can treat hypertension with no or beneficial influence on blood lipids. For smokers, the selective alpha 1-receptor inhibitors may be more attractive, since they also act to counteract the vasoconstriction produced by nicotine. In the future, inhibitors of hydroxymethylglutaryl coenzyme A reductase may offer potential for effective control of blood lipids in hypertensive patients who smoke.


The effect of partially purified dexamethasone receptor on RNA synthesis in rat thymus and liver nuclei.
DEAE Sephadex-A-50 chromatography of rat liver and thymus cytosol charged with dexamethasone (0.1 microM) and heated at 25 degrees C for 30 min, separates two main hormone--receptor complexes, one in the flow through (DE-1) and one eluting with 150 mM [Cl-] (DE-2). Incubation of these two fractions with homologous nuclei results in a 20% stimulation and a 20-25% inhibition of transcription of liver and thymus nuclei, respectively. No difference was observed between the action of DE-1 and DE-2. If the DE-1 and DE-2 fractions of cytosol charged with the glucocorticoid antagonist, cortexolone, are used under the same experimental conditions, no effect on the transcriptional activity of liver or thymus nuclei can be observed. The fact that from nuclei only DE-1 can be isolated, not DE-2, suggests that DE-1 is the intranuclearly active form and that DE-2, with a still unknown mechanism, is transformed to DE-1. A linear relation exists between the number of GR-complexes (DE-1 or DE-2) translocated and the change in RNA synthesis. Transcriptional effects start when a critical number of acceptor sites are occupied in the nucleus (approximately 200-300 sites/nucleus) for both receptor forms.


Stage II/III Rectal Cancer Post-Treatment Surveillance Patterns of Care: A SEER- Medicare Study.
Despite high rectal cancer recurrence rates, knowledge on post-treatment surveillance utilization is limited. Hence, this study aims to estimate patterns of post-treatment surveillance and determine associated factors. Retrospective study of 1,024 SEER-Medicare patients >65 years old diagnosed with stage II/III rectal cancer between 2007-2013. Logistic regression was used to determine factors associated with ≥1 colonoscopy, ≥2 physician visits, ≥2 carcinoembryonic antigen (CEA) tests and ≥2 computed tomographic colonography (CT) within 14 months after primary treatment. Fifty-five percent had ≥1 colonoscopy, 54% had ≥2 physician visits, 47% had ≥2 CEA tests and 20% had ≥2 CTs. In multivariable logistic models, younger age and receipt of chemoradiation therapy (vs none) were significant across all surveillance procedures while clinical factors such as comorbidity were not. Being married (OR=1.69; 95% CI: 1.26-2.26) and proximity to a high-volume hospital (≤15 vs >30 minutes, OR=1.56; 95% CI: 1.00-2.43) were associated with ≥1 colonoscopy. Female gender (OR=1.56; 95% CI: 1.17-2.09), being married (OR=1.56; 95% CI: 1.17-2.08), white race (OR=1.79; 95% CI: 1.23- 2.62) and surgery from high-volume surgeon (OR=1.47; 95% CI: 1.06-2.04) were associated with ≥2 physician visits. Female gender (OR=1.45; 95% CI: 1.08-1.95), being married (OR=1.46; 95% CI: 1.08-1.96) and surgery from high-volume surgeon (OR=1.55; 95% CI: 1.10-2.17) had higher ≥2 CEA tests. Post-treatment surveillance remains low but is more common among younger patients and recipients of chemoradiation. Distinct profiles of patient characteristics and provider volume were associated with individual surveillance procedures suggesting the need for multicomponent strategies to increase surveillance.


Isolation of high-quality total RNA and RNA sequencing of single bovine oocytes.
Studying individual mammalian oocytes has been extremely valuable for the understanding of the molecular composition of oocytes including RNA storage. Here, a detailed protocol for isolation of oocytes, extraction of total RNA from single oocytes followed by full-length cDNA amplification, and library preparation is presented. The procedure permits the production of cost-effective and high-quality sequencing libraries. This protocol can be adapted for transcriptome analysis of oocytes from other species and be used to generate high-quality data from single embryos. For complete details on the use and execution of this protocol, please refer to Biase and Kimble (2018).


Clinical outcomes in international federation of gynecology and obstetrics stage IA endometrial cancer with myometrial invasion treated with or without postoperative vaginal brachytherapy.
To assess the clinical outcomes of patients with Stage IA endometrial cancer with myometrial invasion treated with postoperative vaginal brachytherapy (VBT) with those who received no adjuvant therapy (NAT). All patients treated with hysterectomy for endometrial cancer at Northwestern Memorial Hospital between 1978 and 2005 were identified. Those patients with Stage IA disease with myometrial invasion who were treated with VBT alone or NAT were identified and included in the present analysis. Of 252 patients with Stage IA endometrial cancer with superficial (<50%) myometrial invasion who met the inclusion criteria, 169 underwent VBT and 83 received NAT. The median follow-up in the VBT and NAT groups was 103 and 61 months, respectively. In the VBT group, 56.8% had Grade 1, 37.9% had Grade 2, and 5.3% had Grade 3 tumors. In the NAT group, 75.9%, 20.5%, and 3.6% had Grade 1, 2, and 3 tumors, respectively. Lymphatic or vascular space invasion was noted in 12.4% of the VBT patients and 5.6% of the NAT patients. The 5-year overall survival rate was 95.5%. The 5-year recurrence-free survival rate was 92.4% for all patients, 94.4% for the VBT group, and 87.4% for the NAT group (p = NS). Of the 169 VBT patients and 83 NAT patients, 8 (4.7%) and 6 (7.2%) developed recurrent disease. One vaginal recurrence occurred in the VBT group (0.6%) and three in the NAT group (3.8%). Recurrences developed 2-102 months after surgical treatment. Two of the four vaginal recurrences were salvaged. No Grade 3 or higher acute or late radiation toxicity was noted. The use of postoperative VBT in patients with Stage I endometrial cancer with <50% myometrial invasion yielded excellent vaginal disease control and disease-free survival, with minimal toxicity.


A 308-nm excimer laser for the treatment of scalp psoriasis.
Scalp psoriasis is a frustrating disorder whose treatments can all too often be cumbersome and ineffective. Our objective was to test a combination device involving the 308-nm excimer laser in tandem with a hair blower for treating scalp psoriasis. Adult subjects with scalp psoriasis unresponsive to class I topical steroids used in conjunction with medicated shampoos were treated with 308-nm excimer laser pulses in conjunction with a hair blower that parted the obstructing hair twice a week for up to 15 weeks. Half of the scalp served as a control. Starting doses were based on standard minimal erythema dose (MED)'s with subsequent increments of up to 20%. Thirteen subjects completed the study without adverse events. Two were dropped due to lack of compliance. At the end of the investigation, the difference in the mean modified PASI scores between the control and treated sites was 4.0 (<0.0001). The air blower device in conjunction with the 308-nm excimer laser can safely and effectively treat otherwise refractory scalp psoriasis.


Progressive encephalomyelitis with rigidity and myoclonus (PERM) associated with anti-glycine receptor antibodies and urothelial carcinoma: a case report.
Progressive encephalomyelitis with rigidity and myoclonus (PERM) is a rare neurological condition with paraneoplastic etiology in about 20% of cases, usually presenting before or shortly after the oncological diagnosis is established. PERM associated with anti-glycine receptor antibodies is not previously reported in a patient with bladder cancer. A 72-years-old Caucasian male was admitted with acute onset of dysarthria, dysphagia and trismus three years after initial surgical treatment for bladder cancer. The condition was initially diagnosed as tetanus and treated accordingly, but the diagnosis was reconsidered because of progression despite adequate treatment. Diagnostic workup on readmission revealed lung and paraaortal metastases from bladder cancer and anti-glycine receptor (anti-GlyR) antibodies both in the cerebrospinal fluid and in serum, which supplemented with the clinical presentation led to the diagnosis of PERM, presumably related to bladder cancer. The patient showed improvement and stabilization after treatment with intravenous immunoglobulin and chemotherapy against metastatic bladder cancer. To the best of our knowledge, this is the first reported case of anti-GlyR antibody positive PERM related to urothelial carcinoma. The symptoms mimicked tetanus, and responded to chemotherapy and immunotherapy.


Factors Contributing to Persistent Frequent Attendance in Primary Care Among the Oldest Old: Longitudinal Evidence From the AgeCoDe-AgeQualiDe Study.
Since there is a lack of longitudinal studies in this area, our aim was to identify the determinants of persistent frequent attendance in primary care among the oldest old in Germany. Longitudinal data (follow-up wave 7-9) were taken from the multicenter prospective cohort "Study on needs, health service use, costs, and health-related quality of life in a large sample of oldest-old primary care patients (85+)" (AgeQualiDe), covering primary care patients ≥ 85 years (FU7 n = 741, mean age 88.9 years (SD 2.9; 85-100)). Persistent frequent attenders of general practitioner (GP) services (the patients in the top decile of the number of GP consultations in two or more consecutive waves) were our main outcome of interest. Logistic random-effects models were used. Our analysis included 1,891 observations (766 individuals). Across three waves, we identified 56 persistent frequent attenders. Results of random-effects logistic regressions showed that the odds of being persistent frequent attender were higher for widowed individuals (OR = 4.57; 95% CI [1.07-19.45]). Moreover, a one-point increase in the frailty score and having one more chronic condition increased the odds of being a persistent frequent attender by 68% (OR =1.68; 95% CI [1.05-2.69]) and 23% (OR=1.23, 95% CI [1.05-1.44]), respectively. Our study stressed the longitudinal association between frailty and widowhood as well as chronic diseases and persistent frequent attendance among the oldest old in Germany.


Biocatalytic dechlorination of trichloroethylene with bio-palladium in a pilot-scale membrane reactor.
Trichloroethylene (TCE) is a toxic and recalcitrant groundwater pollutant. An innovative technology using microbial produced Pd(0) nanoparticles for the remediation of TCE contaminated groundwater was developed. The nanoscale bio-Pd particles were precipitated on the biomass of Shewanella oneidensis and hydrogen gas, formate, or formic acid were used as hydrogen donors. Ethane turned out to be the only organic degradation product and no intermediate chlorinated reaction products were detected. Subsequently bio-Pd was implemented in a plate membrane reactor (MR) for the treatment of water containing TCE. In a continuous MR system, containing 50 mg L(-1) bio-Pd, removal rates up to 2,515 mg TCE day(-1) g(-1) Pd were achieved with H(2) gas as hydrogen donor. The measured chloride mass balance confirmed the removal rates. This work shows that a complete, efficient and rapid removal of TCE was achieved with bio-Pd and that a MR system containing bio-Pd and supplied with hydrogen gas offers an alternative for the current remediation technologies of water contaminated with TCE.


A variable number of tandem repeats polymorphism in an E2F-1 binding element in the 5' flanking region of SMYD3 is a risk factor for human cancers.
Histone modification is a crucial step in transcriptional regulation, and deregulation of the modification process is important in human carcinogenesis. We previously reported that upregulation of SMYD3, a histone methyltransferase, promoted cell growth in human colorectal and hepatocellular carcinomas. Here we report significant associations between homozygosity with respect to an allele with three tandem repeats of a CCGCC unit in the regulatory region of SMYD3 and increased risk of colorectal cancer (P = 9.1 x 10(-6), odds ratio = 2.58), hepatocellular carcinoma (P = 2.3 x 10(-8), odds ratio = 3.50) and breast cancer (P = 7.0 x 10(-10), odds ratio = 4.48). This tandem-repeat sequence is a binding site for the transcriptional factor E2F-1. In a reporter assay, plasmids containing three repeats of the binding motif (corresponding to the high-risk allele) had higher activity than plasmids containing two repeats (the low-risk allele). These data suggest that the common variable number of tandem repeats polymorphism in SMYD3 is a susceptibility factor for some types of human cancer.


[Change in the shape and volume of the secretory cells of the mammae in the course of the secretory cycle].
Changes occurring in the acini due to oxytocin were studied using the vital photometry and the morphometric analysis of materials obtained by biopsy. At the initial stages of the functional activation, the secretory cells become cylindric, their volumes simultaneously increasing. A study of intercellular interrelations using the atropine blockade of cholinoreceptors, revealed the lack of activation of the secretory cycle which suggests the functional dissociation of contractile and secretory functions of the cell alveolar complex.


Staufen-2 functions as a cofactor for enhanced Rev-mediated nucleocytoplasmic trafficking of HIV-1 genomic RNA via the CRM1 pathway.
Nucleocytoplasmic shuttling of viral elements, supported by several host factors, is essential for the replication of the human immunodeficiency virus (HIV). HIV-1 uses a nuclear RNA export pathway mediated by viral protein Rev to transport its Rev response element (RRE)-containing partially spliced and unspliced transcripts aided by the host nuclear RNA export protein CRM1. The factor(s) interacting with the CRM1-Rev complex are potential antiretroviral target(s) and could serve as a retroviral model system to study nuclear export machinery adapted by these viruses. We earlier reported that cellular Staufen-2 interacts with Rev, facilitating viral-RNA export. Here, we identified the formation of a complex between Staufen-2, CRM1 and Rev. Molecular docking and simulations mapped the interacting residues in the RNA-binding Domain 4 of Staufen-2 as R336 and R337, which were experimentally verified to be critical for interactions among Staufen-2, CRM1 and Rev by mutational analysis. Staufen-2 mutants defective in interaction with CRM1 or Rev failed to supplement the Rev-RNA export activity and viral production, demonstrating the importance of these interactions. Rev-dependent reporter assays and proviral DNA-construct transfection-based studies in Staufen-2 knockout cells in the presence of leptomycin-B (LMB) revealed a significant reduction in CRM1-mediated Rev-dependent RNA export with decreased virus production as compared to Staufen-2 knockout background or LMB treatment alone, suggesting the relevance of these interactions in augmenting RNA export activity of Rev. Our observations provide further insights into the mechanistic intricacies of unspliced viral-RNA export to the cytoplasm and support the notion that abrogating such interactions can reduce HIV-1 proliferation.


Discrepancies in the concordance of self-reported vision status and visual acuity in the Salisbury Eye Evaluation Study.
To examine the association between self-rated vision and distance visual acuity by race and other factors. Cross-sectional analysis within a longitudinal, population-based cohort study. Two thousand five hundred twenty individuals, 65 to 84 years of age, including 666 black participants and 1854 white participants. All participants reported their self-rated vision score. Binocular distance visual acuity was assessed. Based on the level of visual acuity and the self-reported vision score, participants were placed into concordant and discrepant groups. Multinomial logistic regression models were used to examine the characteristics associated with concordant and discrepant groups. Black participants were more likely to be represented in the discordant group that reported good vision but had acuity worse than 20/40. In the multivariate analysis, a per-year decrement in years of education received increased the odds of being in both discrepant groups, one that reported good vision but had a visual acuity worse than 20/40 (odds ratio, 1.21; P<0.0001) and the other discrepant group that reported bad vision but had a visual acuity of 20/40 or better (odds ratio, 1.04; P<0.0001). A per-year decrement in years of education received also seemed to explain the excess risk of black race in the discrepant group that reported good vision but had a visual acuity worse than 20/40. Given the socioeconomically driven discrepancies in self-reported vision status, self-reported vision status should be used cautiously in surveillance surveys, especially when assessing vision inequalities between socioeconomic groups. The author(s) have no proprietary or commercial interest in any materials discussed in this article.


Extent of altered white matter in unilateral and bilateral periventricular white matter lesions in children with unilateral cerebral palsy.
To investigate the extent of white matter damage in children with unilateral cerebral palsy (UCP) caused by periventricular white matter lesions comparing between unilateral and bilateral lesions; and to investigate a relationship between white matter microstructure and hand function. Diffusion MRI images from 46 children with UCP and 18 children with typical development (CTD) were included. Subjects were grouped by side of hemiparesis and unilateral or bilateral lesions. A voxel-wise white matter analysis was performed to identify regions where fractional anisotropy (FA) was significantly different between UCP groups and CTD; and where FA correlated with either dominant or impaired hand function (using Jebsen Taylor Hand Function Test). Children with unilateral lesions had reduced FA in the corticospinal tract of the affected hemisphere. Children with bilateral lesions had widespread reduced FA extending into all lobes. In children with left hemiparesis, impaired hand function correlated with FA in the contralateral corticospinal tract. Dominant hand function correlated with FA in the posterior thalamic radiations as well as multiple other regions in both left and right hemiparesis groups. Periventricular white matter lesions consist of focal and diffuse components. Focal lesions may cause direct motor fibre insult resulting in motor impairment. Diffuse white matter injury is heterogeneous, and may contribute to more global dysfunction.


Central venous oxygen saturation for the control of automatic rate-responsive pacing.
A rate-adjusting pacemaker system is described which uses central venous oxygen saturation (SO2) for the regulation of the pacing rate. This system was tested externally in ten patients with chronically implanted VVI units. There was an average increase in cardiac output of 18% with the SO2-regulated pacemaker as compared to the situation during fixed rate stimulation. Central venous oxygen saturation appears to be an ideal biological parameter for autoregulating the pacing rate. It represents the only sensor suitable for the realization of a closed feedback loop concept.


Splenic Artery Embolization for Treatment of Refractory Ascites After Liver Transplantation.
Post-transplantation refractory ascites is uncommon; however, it can be a serious problem, increasing both morbidity and mortality in patients. Despite scant literature available, splenic artery embolization (SAE) has been shown to be an effective treatment for refractory ascites after cadaveric orthotopic liver transplantation (OLT). We report a successful use of therapeutic SAE for refractory ascites post-OLT.


Comparing sustainable product hashtags: Insights from a historical twitter dataset.
This data article describes the process of data collection and analysis of Twitter conversations about sustainable products. The dataset contains the IDs of tweets tagged with the hashtags #sustainableproducts, #ecoproducts, #ecofriendlyproducts, and #greenproducts. The time period spans 10 years and includes a total of over 140 thousand tweets from around the world. The article describes the process of obtaining the data using Twarc and the Twitter developer's academic researcher API and describes the preprocessing techniques used to identify keywords, hashtags, topics, and sentiments expressed in the conversations. The analysis identifies key attributes of each sustainable product category as well as commonalities and differences within and across categories. The data have the potential to be reused in future research related to sustainable consumption and production, including further analysis of the sentiments and attitudes expressed in the Twitter conversations and comparison with other social media platforms or survey data. In addition, the data can serve as a basis for marketing strategies and product design by enterprises or organizations seeking to promote sustainable products.


Coincidence of two novel arylsulfatase A alleles and mutation 459+1G>A within a family with metachromatic leukodystrophy: molecular basis of phenotypic heterogeneity.
In a family with three siblings, one developed classical late infantile metachromatic leukodystrophy (MLD), fatal at age 5 years, with deficient arylsulfatase A (ARSA) activity and increased galactosylsulfatide (GS) excretion. The two other siblings, apparently healthy at 12(1/2) and 15 years, respectively, and their father, apparently healthy as well, presented ARSA and GS values within the range of MLD patients. Mutation screening and sequence analysis disclosed the involvement of three different ARSA mutations being the molecular basis of intrafamilial phenotypic heterogeneity. The late infantile patient inherited from his mother the frequent 0-type mutation 459+1G>A, and from his father a novel, single basepair microdeletion of guanine at nucleotide 7 in exon 1 (7delG). The two clinically unaffected siblings carried the maternal mutation 459+1G>A and, on their paternal allele, a novel cytosine to thymidine transition at nucleotide 2435 in exon 8, resulting in substitution of alanine 464 by valine (A464V). The fathers genotype thus was 7delG/A464V. Mutation A464V was not found in 18 unrelated MLD patients and 50 controls. A464V, although clearly modifying ARSA and GS levels, apparently bears little significance for clinical manifestation of MLD, mimicking the frequent ARSA pseudodeficiency allele. Our results demonstrate that in certain genetic conditions MLD-like ARSA and GS values need not be paralleled by clinical disease, a finding with serious diagnostic and prognostic implications. Moreover, further ARSA alleles functionally similar to A464V might exist which, together with 0-type mutations, may cause pathological ARSA and GS levels, but not clinical outbreak of the disease.


Consistency assessment and visualization of recommendations on gastroesophageal reflux disease: a scoping review of clinical practice guidelines.
Gastroesophageal reflux disease (GERD) is a highly prevalent gastrointestinal disorder that causes diverse esophageal and extraesophageal symptoms. Many clinical practice guidelines (CPGs) have been issued around the world to provide practical evidence regarding GERD management. However, some of the recommendations discussed in various CPGs are inconsistent across individual documents. We aimed to summarize the evidence from CPGs on GERD and assess the consistency of the recommendations. In this scoping review, we identified current CPGs on the clinical management of GERD, which were comprehensively searched for in electronic databases and on relevant scientific websites. The recommendations were extracted using the population‑intervention‑comparison framework and were subsequently categorized into tables. Ultimately, 24 CPGs were identified. They included 86 recommendations, which were classified into 5 categories: definition, epidemiology, diagnosis, treatment, and complications. Among the identified recommendations, 68 were proposed in at least 2 CPGs, and they were assessed for the consistency of direction and strength. Overall, 32.4% of the recommendations (22/68) were consistent in direction and strength, whereas 60.3% (41/68) were consistent in direction but inconsistent in strength. Moreover, 7.4% (5/68) were inconsistent in direction. These referred to the relationship between GERD and tobacco consumption, Helicobacter pylori infection, diagnostic utility of the 2‑week proton pump inhibitor test, cessation of special food, and antireflux surgery for GERD with extraesophageal symptoms. Most CPG recommendations regarding GERD were consistent in direction, except for 5 discrepancies, for which further well‑designed, large‑scale research is required to explain the inconsistency.


EEG-based methods for recovery prognosis of patients with disorders of consciousness: A systematic review.
Disorders of consciousness (DoC) are acquired conditions of severely altered consciousness. Electroencephalography (EEG)-derived biomarkers have been studied as clinical predictors of consciousness recovery. Therefore, this study aimed to systematically review the methods, features, and models used to derive prognostic EEG markers in patients with DoC in a rehabilitation setting. We conducted a systematic literature search of EEG-based strategies for consciousness recovery prognosis in five electronic databases. The search resulted in 2964 papers. After screening, 15 studies were included in the review. Our analyses revealed that simpler experimental settings and similar filtering cut-off frequencies are preferred. The results of studies were categorised by extracting qualitative and quantitative features. The quantitative features were further classified into evoked/event-related potentials, spectral measures, entropy measures, and graph-theory measures. Despite the variety of methods, features from all categories, including qualitative ones, exhibited significant correlations with DoC prognosis. Moreover, no agreement was found on the optimal set of EEG-based features for the multivariate prognosis of patients with DoC, which limits the computational methods applied for outcome prediction and correlation analysis to classical ones. Nevertheless, alpha power, reactivity, and higher complexity metrics were often found to be predictive of consciousness recovery. This study's findings confirm the essential role of qualitative EEG and suggest an important role for quantitative EEG. Their joint use could compensate for their reciprocal limitations. This study emphasises the need for further efforts toward guidelines on standardised EEG analysis pipeline, given the already proven role of EEG markers in the recovery prognosis of patients with DoC.


The in vivo metabolites of [14C] progesterone in bovine muscle and adipose tissue.
Muscle and adipose tissue were obtained from steers and dairy cows following subcutaneous administration of [14C] progesterone. Following extraction, purification and separation by column, thin layer and gas-liquid chromatography, various radioactive residues from these tissues were identified by their chromatographic mobility, crystallization to constant specific activity and mass spectra. Progesterone constituted 54% of free radioactivity extracted from muscle and 69 and 73% of radioactivity in the free and conjugated portions of extracts, respectively, from fat. Metabolites identified were: 5alpha-pregnane-3,20-dione, 9%, 0%, 0%, 20beta-hydroxy-4-pregnen-3-one, 8%, 11%, 3%; 3alpha-hydroxy-5beta-pregnan-20-one, 13%, 2%, 2%, 3alpha-hydroxy-5alpha-pregnan-20one, 3%, 3%, 6%; 20 alpha-hydroxy-5alpha-pregnan-3-one, 0%, 2%, 3%; of radioactivity in muscle (free) and fat (free and conjugated fractions), respectively. Tentatively identified in fat extracts by chromatographic mobility were: 20alpha-hydroxy-4-pregnen-3-one, 1%, 1% and 3beta-hydroxy-5beta-pregnan-20-one, 0%, 2% of radioactivity in free and conjugated fractions, respectively. The average concentration of steroid in these animals due solely to treatment, calculated from the specific activity of the [14C] progesterone administered, was 3.4 and 18.1 ng/g in muscle and subcutaneous fat, respectively.


Variable absorption of carbidopa affects both peripheral and central levodopa metabolism.
Carbidopa (CD), a competitive inhibitor of aromatic l-amino acid decarboxylase that does not cross the blood-brain barrier, is routinely administered with levodopa (LD) to patients with Parkinson disease (PD) to reduce the peripheral decarboxylation of LD to dopamine. Using a stable isotope-labeled form of LD, the authors examined in 9 PD patients the effects of variable CD absorption on peripheral and central LD metabolism. Subjects were administered orally 50 mg of CD followed in 1 hour by a slow bolus intravenous infusion of 150 mg stable isotope-labeled LD (ring 1',2',3',4',5',6'-13C). Eight patients underwent a lumbar puncture 6 hours following the infusion. Blood and cerebrospinal fluid (CSF) samples were analyzed for labeled and unlabeled metabolites using a combination of high-performance liquid chromatography and mass spectrometry. When patients were divided into "slow" and "rapid" CD absorption groups, significantly greater peripheral LD decarboxylation (as measured by area under the curve [AUC]-labeled serum HVA) was noted in the poor absorbers (p = 0.05, Mann-Whitney U test). Elimination half-lives for serum LD did not differ between groups, suggesting a further capacity for decarboxylation inhibition in the "rapid" absorbers. A significant correlation between AUC serum CD and percent-labeled HVA in CSF was found for all patients (R = 0.786, p = 0.02). "Rapid" as compared to "slow" CD absorbers had significantly more percent-labeled CSF HVA (60 vs. 49, p = 0.02, Mann-Whitney U test), indicating greater central-labeled DA production in the better CD absorbers. The data suggest that peripheral aromatic l-amino acid decarboxylase activity is not saturated at CD doses used in current practice. The authors believe that future studies to better examine a dose dependence of CD on peripheral LD decarboxylation and LD brain uptake are warranted.


[Expression of the gene encoding human leukocyte differentiation antigen 5D4].
To make sure that 5D4 cDNA is a full length cDNA and to prepare antigens to make monoclonal antibodies against different epitopes in 5D4 protein. Recombinant expression vectors pGEX-5X-3-5D4 (full length) and pGEX-5X-3-5D4 (5'fragment) were constructed. 5D4 polypeptides were expressed in Escherichia coli DH5 alpha. 5D4 mRNA expression was detected in various tissues and cell lines with Northern blot assay and RT-PCR. Northern blot demonstrated five mRNA transcripts of 1.9, 2.4, 3.9, 4.5, and 10.0 kb in length existing and expressing of various extent in various tissues. The 1.9 kb mRNA transcript was observed only in the spleen and the small intestinal tissues. The expression of 5D4 mRNA was different in various cell lines. 5D4 polypeptides were expressed in Escherichia coli DH5 alpha. The 1,846 bp 5D4 cDNA is a full length cDNA. A full length 5D4 protein and a 5D4 polypeptide which is the NH2 side to the transmembrane region fused with GST with a molecular weight of 66,000 and 60,000 expressing in Escherichia coli DH5 alpha.


CD137 deficiency because of two novel biallelic TNFRSF9 mutations in a patient presenting with severe EBV-associated lymphoproliferative disease.
Increasing evidence indicates that some germline genetic mutations that impair pathways required for robust host immune surveillance against EBV infection may result in an extremely high susceptibility to EBV-associated lymphoproliferative disease (EBV+ LPD). TNFRSF9 encodes a vital costimulatory molecule that enhances CD8+ T-cell proliferation, survival and cytolytic activity. To date, no relevant case resulting from TNFRSF9 heterozygous mutations has been identified. Here, we report the first case of CD137 deficiency caused by two novel biallelic heterozygous TNFRSF9 mutations [NM_001561.5: c.208 + 1->AT and c.452C>A (p.T151K)] in a patient presenting with severe EBV+ LPD. Immunophenotyping and in vitro assays of lymphocyte function and NK cell activity were performed. Biallelic TNFRSF9 mutations resulted in markedly reduced or abrogated expression of CD137 on activated T, B and NK cells. CD8+ T cells from the patient had impaired activation, reduced expression/release of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), perforin and granzyme B, and diminished cytotoxic activity. Functional experiments identified both variations were hypomorphic mutations and played a contributing role in CD137 deficiency and the development of EBV+ LPD. Our study expands the genetic spectrum and clinical phenotype of patients with CD137 deficiency and provides additional evidence that the TNFRSF9 gene plays a critical role in host immune responses to EBV infection.


Light and electron microscopic immunocytochemical demonstration of intraepidermal CGRP-containing nerves in human skin.
The distribution of nerves immunoreactive to calcitonin gene-related peptide (CGRP) was studied in human skin with special reference to the intraepidermal nerve fibres. It was shown by light and electron microscopy that a small proportion of the CGRP immunoreactive nerves entered the epidermis. The majority of the nerves ended as free nerve terminals subepidermally. Intraepidermal nerve endings were mainly seen in skin areas, which have a dense sensory innervation, such as fingertips and foreskin. The role of the peptidergic nerves in human skin is discussed.


A problem with clinical T factor in the 8th TNM edition: Prognosis and EGFR mutation status of small sized lung cancers with difficulty to measure the diameter of solid component in part-solid tumor.
Clinical T factors in the 8th TNM classification of lung cancer have a practical problem. In some cases, it is difficult to measure the size of the solid components in part-solid tumors, and the classification of these tumors is controversial. We evaluated 590 resected cT1N0M0 stage IA non-small-cell lung cancers based on the 7th edition between 2009 and 2012. Tumor and solid component diameters were measured using thin-section computed tomography (CT). We defined tumors with difficulty in measuring the size of the solid components as lung cancers with scattered or mixed consolidation (LCSMCs). LCSMCs were observed in 79 (13.4%) patients. Other tumors were classified as cTis, cT1mi, cT1a, cT1b, and cT1c, according to the 8th edition. We compared prognosis and epidermal growth factor receptor mutations (EGFRm) status of LCSMCs with those of cT1a, cT1b, and cT1c. The difference in overall survival (OS) among cT1a, cT1b, and cT1c was significant (5-year-OS: 96.9% vs. 76.8% vs. 65.0%). There was no significant difference in prognosis between LCSCs and cT1a (5-year-OS: 92.4% vs. 96.9%). A significant difference was observed in the frequency of EGFRm between cT1a, cT1b, and cT1c (52.4%, 42.4%, and 29.8%). The incidence of EGFRm in LCSMCs was 54.8% and there was no significant difference between LCSMCs and cT1a. The prognosis and frequency of EGFRm in LCSMCs were close to those in cT1a. As we cannot measure the diameter of the solid component in subsolid lung cancers, it may be appropriate to classify these tumors as cT1a tumors.


Inter- and intra-observer reliability of a smartphone application for measuring hallux valgus angles.
Measurement of radiological angles can be useful in the planning of the management of patients with hallux valgus. A smartphone application offers an alternative way of measuring these angles in a clinic setting. We compared the reliability (inter- and intra-observer) of this method to the use of PACS. Radiographs of 30 feet from new patients referred with hallux valgus were examined and angles (HVA, IMA, and DMAA) recorded using the smartphone application and PACS. The smartphone application provided good inter-observer reliability for HVA and IMA (r=0.93 and r=0.79 respectively). Intra-observer reliability for HVA and IMA was also found to be good (r=0.93-0.97 r=0.82-0.93 respectively). The inter- and intra-observer reliability for using this method to measure DMAA fell below useful levels (r<0.60 in each case). This smartphone application provides a reliable method to measure HVA and IMA but we would not recommend it to measure DMAA.


Enhanced photocatalytic hydrogen generation over ZrO2-TiO2-CdS hybrid structure.
Hybrid photocatalysts with suitable band structures are expected to show enhanced photocatalytic activity as compared to their constituent single phase compounds due to their improved physico-chemical properties. Here, we report an enhanced photocatalytic activity of a new composite photocatalyst comprising of ZrO2, TiO2, and CdS. This hybrid catalyst exhibits increased photocatalytic activity for hydrogen generation from water as compared to their constituent compounds. The photocatalytic activity decreases in the order: ZrO2-TiO2-CdS>TiO2-CdS>ZrO2-CdS>CdS>ZrO2-TiO2≈TiO2>ZrO2. An apparent quantum efficiency of 11.5% is obtained for ZrO2-TiO2-CdS with Pd as co-catalyst. Absorption edge of the composite is slightly blue shifted compared with that of pure CdS. Photoluminescence lifetime studies indicate an increased lifetime for the charge carriers in the composite sample as compared to that of pure CdS. Transmission electron microscopy images reveal that the particle size of the composite is much less than that of single phase CdS. The enhanced photocatalytic activity of the composite is attributed to the decreased particle size of CdS and increased lifetime of the charge carriers resulting from the efficient interfacial transfer of photogenerated electrons at the CdS/TiO2 and CdS/ZrO2 interface.


Suppression of pancreatic tumor growth by targeted arsenic delivery with anti-CD44v6 single chain antibody conjugated nanoparticles.
Arsenic trioxide (As2O3) is a promising anticancer agent for solid tumors. However, the high toxicity to normal tissues resulting from the lack of tumor specificity remains a huge challenge in its systemic application. Targeted vectors enabling drug delivery to specific cancer cells bring about great potential for better therapeutic efficacy whereas low side effects in cancer treatments. Our previous work has demonstrated that the anti-CD44v6 single chain variable fragment (scFv(CD44v6)) screened out from the human phage-displayed scFv library possesses high specificity and affinity to membrane antigen CD44v6 over-expressing in a subset of epithelium-derived cancers, such as pancreatic, hepatocellular, colorectal and gastric cancers. Herein, a maleimide-functionalized amphiphilic diblock copolymer of poly (ethylene glycol) and poly (D, L-lactide) (mal-PEG-PDLLA) was synthesized and assembled to vesicles with arsenite ion (As) encapsulated in their cores (As-NPs). Conjugation of scFv(CD44v6) with mal-PEG-PDLLA (scFv-As-NPs) enabled more efficient delivery of As and exhibited higher cytotoxic activity than non-targeted ones (As-NPs) in human pancreatic cancer cells PANC-1. Furthermore, the targeted delivery of As induced more significant gene suppression in terms of the expression of anti-apoptotic Bcl-2 protein. Consequently, the expression level of cleaved caspase-3 which is a molecular indicator of cell apoptosis was remarkably elevated. In animal tests, scFv-As-NPs were found to greatly increase accumulation of drug in tumor site and potentiate the efficacy of As in inhibiting tumor growth owing to the enhanced cell apoptosis. These results imply that our tumor specific nanocarriers provide a highly efficient and safe platform for pancreatic cancer therapy.


Extracellular acidosis and very low [Na+ ] inhibit NBCn1- and NHE1-mediated net acid extrusion from mouse vascular smooth muscle cells.
The electroneutral Na+ , HCO3- cotransporter NBCn1 and Na+ /H+ exchanger NHE1 regulate acid-base balance in vascular smooth muscle cells (VSMCs) and modify artery function and structure. Pathological conditions - notably ischaemia - can dramatically perturb intracellular (i) and extracellular (o) pH and [Na+ ]. We examined effects of low [Na+ ]o and pHo on NBCn1 and NHE1 activity in VSMCs of small arteries. We measured pHi by 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein-based fluorescence microscopy of mouse mesenteric arteries and induced intracellular acidification by NH4+ prepulse technique. NBCn1 activity - defined as Na+ -dependent, amiloride-insensitive net base uptake with CO2 /HCO3- present - was inhibited equally when pHo decreased from 7.4 (22 mm HCO3-/5% CO2 ) by metabolic (pHo 7.1/11 mm HCO3-: 22 ± 8%; pHo 6.8/5.5 mm HCO3-: 61 ± 7%) or respiratory (pHo 7.1/10% CO2 : 35 ± 11%; pHo 6.8/20% CO2 : 56 ± 7%) acidosis. Extracellular acidosis more prominently inhibited NHE1 activity - defined as Na+ -dependent net acid extrusion without CO2 /HCO3- present - at both pHo 7.1 (45 ± 9%) and 6.8 (85 ± 5%). Independently of pHo , lowering [Na+ ]o from 140 to 70 mm reduced NBCn1 and NHE1 activity <20% whereas transport activities declined markedly (25-50%) when [Na+ ]o was reduced to 35 mm. Steady-state pHi decreased more during respiratory (ΔpHi /ΔpHo = 71 ± 4%) than metabolic (ΔpHi /ΔpHo = 30 ± 7%) acidosis. Extracellular acidification inhibits NBCn1 and NHE1 activity in VSMCs. NBCn1 is equivalently inhibited when pCO2 is raised or [HCO3-]o decreased. Lowering [Na+ ]o inhibits NBCn1 and NHE1 markedly only below the typical physiological and pathophysiological range. We propose that inhibition of Na+ -dependent net acid extrusion at low pHo protects against cellular Na+ overload at the cost of intracellular acidification.


Characterization of the anti-fungal activity of a Bacillus spp. associated with sclerotia from Sclerotinia sclerotiorum.
Sclerotinia sclerotiorum fruiting bodies (sclerotia) were found to harbour bacteria that possess anti-fungal activity. Among 1,140 bacterial isolates collected, 32 were found to inhibit the growth of four common fungal pathogens of canola, S. sclerotiorum, Rhizoctonia solani, Alternaria brassicae and Leptosphaeria maculans. One of these broad-spectrum isolates, LEV-006, was found to be closely related to Bacillus subtilis based on 16S rRNA analysis. The anti-fungal activities were purified and found to be associated with a low molecular weight peptide complex consisting mostly of the cyclic lipopeptide fengycin A and B, as revealed by matrix-assisted laser desorption/ionization time-of-flight and post-source decay analysis, as well as two proteins of 20 and 55 kDa. Peptide mass fingerprinting revealed that the 55-kDa protein was similar to vegetative catalase 1; however, when the enzyme was expressed in Escherichia coli, it exhibited catalase but not anti-fungal activity. The sequences of several peptides from the 20-kDa protein were obtained and indicated that it was a unique anti-fungal protein.


Clinical Significance of Tumor Deposits in Gastric Cancer: a Retrospective and Propensity Score-Matched Study at Two Institutions.
Gastric cancer (GC) treatment is largely determined by tumor stage. Despite improvements in the mode of treatment of various types of advanced disease, staging is still evolving. The role of tumor deposits (TDs) in staging remains debated. The purpose of this research is to investigate the relationship between TDs and prognosis in GC. A total of 3098 patients were considered eligible for prognostic analysis (2706 patients in the TDs-negative group and 392 patients in the TDs-positive group). A one-to-one propensity score-matching analysis was performed using a logistic regression mode and the following covariates: age, gender, tumor location, size, differentiation, perineural invasion, lymphovascular invasion, pTNM stage, type of gastrectomy, and the number of lymph nodes retrieved between TDs-negative and TDs-positive group, then 323 patients in each group were analyzed. Univariate and multivariate analyses of prognostic factors were conducted accordingly. The predictive ability of different staging system incorporating TDs was evaluated. TDs were present in 14.5% cases and almost all of the patients (99%) suffered from advanced GC. Multivariate analysis showed that pN stage, chemotherapy, and TDs were the independent prognostic factors. The TDs-positive group showed a lower rate of 5-year disease-free survival compared with the TDs-negative group in all patients, stage II, and stage III patients (p = 0.001, 0.029, and 0.003, respectively). The 5-year disease-free survival for patients with TDs and without TDs was 27.6% and 34.4%, respectively. Our research shows that TDs are closely associated with prognosis in GC. TDs should be incorporated into the TNM staging system, which could then accurately improve the staging reliability and prognostic assessment.


The investigation of storage situation of fish muscle via the analysis of its exudate by MALDI-TOF MS.
The potential of MALDI-TOF MS was investigated in terms of its capability to determine the change of exudates of fish muscle with different freeze-thaw cycles (0, 1 and 2), and frozen storage periods. The exudates were collected from dead chilled marine fish species, large yellow croacker (Larimichthys crocea, LC) and freshly slaughtered freshwater fish species, Japanese seabass (Lateolabrax japonicus) to be studied as models. 109 proteins, in which, 32 are extracellular proteins, and 15 are intracellular proteins, were identified by analyzing exudate of LC using MALDI-TOF MS and HPLC-MS/MS. The results show that the present method may be able to determine the change of fish muscle foods in a more sensitive mode than K value indicated quality control. The feasibility of verifying the storage situation of fish sample was performed by analyzing fish samples obtained from the local market. It is promising to estimate the storage situation of fishery products or other animal muscle foods by analyzing their muscle exudates based on the presently developed strategy.


The valance state of vanadium-key factor in the flexibility of potassium vanadates structure as cathode materials in Li-ion batteries.
Potassium hexavanadate (K2V6O16·nH2O) nanobelts have been synthesized by the LPE-IonEx method, which is dedicated to synthesis of transition metal oxide bronzes with controlled morphology and structure. The electrochemical performance of K2V6O16·nH2O as a cathode material for lithium-ion batteries has been evaluated. The KVO nanobelts demonstrated a high discharge capacity of 260 mAh g-1, and long-term cyclic stability up to 100 cycles at 1 A g-1. The effect of the vanadium valence state and unusual construction of the nanobelts, composed of crystalline and amorphous domains arranged alternately were also discussed in this work. The ex-situ measurements of discharged electrode materials by XRD, MP-AES, XAS and XPS show that during the subsequent charge/discharge cycle the potassium in the K2V6O16·nH2O structure are replacing by lithium. The structural stability of the potassium hexavandate during cycling depends on the initial vanadium valence state on the sample surface and the presence of the "fringe free" domains in the K2V6O16·nH2O nanobelts.


HSF and Hsp Gene Families in sunflower: a comprehensive genome-wide determination survey and expression patterns under abiotic stress conditions.
Sunflowers belong to the Asteraceae family, which comprises nutrimental and economic oilseed plants. Heat shock proteins (Hsps) are protein families vital for all organisms' growth and survival. Besides the ordinary conditions, the expression of these proteins ascends during abiotic stress factors such as high temperature, salinity, and drought. Using bioinformatics approaches, the current study identified and analyzed HSF and Hsp gene family members in the sunflower (Helianthus annuus L.) plant. HSF, sHsp, Hsp40, Hsp60, Hsp70, Hsp90, and Hsp100 domains were analyzed in the sunflower genome, and 88, 72, 192, 52, 85, 49, and 148 genes were identified, respectively. The motif structures of the proteins in the same phylogenetic tree were similar, and the α-helical form was dominant in all the protein families except for sHsp. The estimated three-dimensional structure of 28 sHsp proteins was determined as β-sheets. Considering protein-protein interactions, the Hsp60-09 protein (38 interactions) was found to be the most interacting protein. The most orthologous gene pairs (58 genes) were identified between Hsp70 genes and Arabidopsis genes. The expression analysis of selected genes was performed under high temperature, drought, and high temperature-drought combined stress conditions in two sunflower cultivars. In stress conditions, gene expressions were upregulated for almost all genes in the first half and first hours at large. The expressions of HanHSF-45 and HanHsp70-29 genes were raised in two cultivars under high temperature and high temperature-drought combined stress conditions. This study presents a blueprint for subsequent research and delivers comprehensive knowledge of this vital protein domain.


Fouling characterization and control for harvesting microalgae Arthrospira (Spirulina) maxima using a submerged, disc-type ultrafiltration membrane.
This study characterized the fouling of a novel circular-disc ultrafiltration membrane in a submerged bioreactor system to harvest Arthrospira maxima cells. Flux-stepping study showed that the value of critical flux was below the smallest flux tested at 28.8lm(-2)h(-1), and that the membrane was to operate above the critical flux to sustain the necessary rate of cell concentration. The membrane with similar pore size but greater pore density experienced not only lesser degree of total resistance, but also possessed smaller fraction of irreversible resistance. Membrane fouling was mainly attributed to fragmented cells rather than to soluble or extracellular polymeric substances. Furthermore, flux recovery studies demonstrated that membrane relaxation and surface cleaning could partially recover fluxes for both low (6gl(-1)) and high (40gl(-1)) cell densities, whereas backwashing could fully recover fluxes. Calculation of energy consumption and cell harvesting productivity also favoured membrane filtration with backwashing.


Production of thermostable pectinase and xylanase for their potential application in bleaching of kraft pulp.
A very high level of alkalophilic and thermostable pectinase and xylanase has been produced from newly isolated strains of Bacillus subtilis and Bacillus pumilus respectively. Enzyme production for pectinase was carried out under SSF using combinations of cheap agricultural residues while xylanase was produced under submerged fermentation using wheat bran as substrate to minimize the cost of production of these enzymes Among the various substrates tested, the highest yield of pectinase production was observed by using combination of WB + CW (6592 U/g of dry substrate) supplemented with 4% yeast extract when incubated at 37 degrees C for 72 h using deionized water of pH 7.0 as moistening agent. The biobleaching effect of these cellulase free enzymes on kraft pulp was determined. Both xylanase and pectinase showed stability over a broad range of pH from 6 to 10 and temperature from 55 to 70 degrees C. The bleaching efficiency of the pectinase and xylanase on kraft pulp was maximum after 150 min at 60 degrees C using enzyme dosage of 5 IU/ml of each enzyme at 10% pulp consistency with about 16% reduction in kappa number and 84% reduction in permanganate number. Enzyme treated pulp when subjected to CDED(1)D(2) steps, 25% reduction in chlorine consumption and upto 19% reduction in consumption of chlorine dioxide was observed for obtaining the same %ISO brightness. Also an increase of 22 and 84% in whiteness and fluorescence respectively and a decrease of approximately 19% in the yellowness of the biotreated pulp were observed by pretreatment of the pulp with our enzymatic mixture.


Binge Drinking, Non-injection Drug Use, and Sexual Risk Behaviors among Adolescent Sexual Minority Males, 3 US Cities, 2015.
In 2016, more than 90% of HIV diagnoses among young men aged 13-19 years were attributed to male-male sexual contact. Little is known about how binge drinking and non-injection drug use may be associated with risky sexual behavior among adolescent sexual minority males (ASMM). Using data from the National HIV Behavioral Surveillance, we examined how binge drinking and non-injection drug use may be associated with sexual risk among ASMM. ASMM were recruited for interviews in 3 cities: Chicago, New York City, and Philadelphia. Among 16-18-year-olds (N = 488), we evaluated the association between binge drinking (≥ 5 drinks in one sitting in the past 30 days), non-injection drug use (past 12-month use of methamphetamines, powder cocaine, downers, painkillers, ecstasy, poppers, and "other"), and two past 12-month sexual risk outcomes: condomless anal intercourse with a casual partner and having multiple sex partners. We used log-linked Poisson regression models with robust standard errors to estimate prevalence ratios (PR) and 95% confidence intervals (CI). Overall, 26% of 16-18-year-old ASMM binge drank, and 21% reported non-injection drug use. Among ASMM who binge drank, 34% reported condomless anal intercourse with a casual partner compared with 22% of those who did not (PR: 1.53, 95% CI: 1.04-2.26). Similarly, 84% of ASMM who binge drank reported having multiple partners compared with 61% of those who did not (PR: 1.38, 95% CI: 1.09-1.76). Among ASMM who used non-injection drugs, 37% reported condomless anal intercourse compared with 22% of those who did not (PR: 1.70, 95% CI 1.09-2.50), while 86% of those who used non-injection drugs had multiple partners compared with 62% of those who did not (PR: 1.40, 95% CI: 1.06-1.80). Our findings suggest that the prevalence of substance misuse is high among sexual minority youth and is associated with sexual risk in this population. Our findings highlight the need for high-quality HIV prevention programs for ASMM especially as HIV prevention programs for this population are scarce.


The genetics of tritan disturbances.
Tritan (blue-green) colour vision disturbances have been found in 79 individuals in six families, revealing an autosomal dominant mode of inheritance with a wide variability of test results within families. Evidence is presented that it is--in contradistinction to the X-chromosomally inherited red-green defects--incorrect to make a subdivision between dichromasia (tritanopia) and anomalous trichromasia (tritanomaly). On the basis of three small screening series, totalling 1900 individuals, the frequency of tritan disturbances is estimated to be around 2 per 1000. Seven males have been observed carrying both inherited tritan and red-green defects.


The Relationship of Ready-to-Eat Cereal Intake and Body Weight in Adults: A Systematic Review of Observational Studies and Controlled Trials.
Results from observational studies indicate that consumption of ready-to-eat cereal (RTEC) is associated with higher diet quality and lower incidence of overweight and obesity in adults compared with other breakfasts or skipping breakfast. However, randomized controlled trials (RCTs) have had inconsistent results regarding effects of RTEC consumption on body weight and composition. This systematic review aimed to evaluate the effect of RTEC intake on body weight outcomes in observational studies and RCTs in adults. A search of PubMed and Cochrane Central Register of Controlled Trials (CENTRAL) databases yielded 28 relevant studies, including 14 observational studies and 14 RCTs. Results from observational studies demonstrate that frequent RTEC consumers (usually ≥4 servings/wk) have lower BMI, lower prevalence of overweight/obesity, less weight gain over time, and less anthropometric evidence of abdominal adiposity compared with nonconsumers, or less frequent consumers. RCT results suggest that RTEC may be used as a meal or snack replacement as part of a hypocaloric diet, but this approach is not superior to other options for those attempting to achieve an energy deficit. In addition, RTEC consumption was not associated with significantly less loss of body weight, or with weight gain, in any of the RCTs. RTEC intake is associated with favorable body weight outcomes in adults in observational studies. RTEC does not hinder weight loss when used as a meal or snack replacement within a hypocaloric diet. Additional long-term RCTs (≥6 mo) in both hypocaloric and ad libitum conditions are recommended to evaluate further the potential effects of RTEC consumption on body weight outcomes. PROSPERO (CRD42022311805).


Inducible nitric oxide synthase and argininosuccinate synthetase: co-induction in brain tissue of patients with Alzheimer's dementia and following stimulation with beta-amyloid 1-42 in vitro.
In Alzheimer's disease (AD), amyloid plaques within the brain are surrounded by activated glial cells (microglia and astrocytes). The mechanisms of glial activation and its effect on disease progression are not fully understood. Growing evidence suggests that beta-amyloid (Abeta) peptide, a major constituent of the amyloid plaque, is critically involved in the induction of an inflammatory response. The goal of this study was to examine the role of Abeta in the pathogenesis of local inflammation and neuronal cell death. We found increased mRNA levels of inducible nitric oxide synthase (iNOS) and the arginine regenerating enzyme argininosuccinate synthetase (ASS) within cortices of AD patients suggesting high output NO production. In vitro, synthetic Abeta1-42 and to a lesser extent Abeta1-40 induced iNOS and ASS transcription with consecutive NO overproduction in mixed rat neuronal-glial cultures. Furthermore, Abeta-stimulation lead to an increased release of inflammatory cytokines interleukin (IL)-1beta, IL-6 and tumor necrosis factor-alpha. Again, Abeta1-42 had a much more pronounced effect as compared to Abeta1-40. Our data suggest that Abeta1-42 is a key mediator of glial activation and via the induction of inflammatory mediators may be a critical component of the neurodegenerative process in AD.


A comparison of the 'Sheffield' Prognostic Index with forearm muscle dynamometry in patients from Sheffield undergoing major abdominal and urological surgery.
An increased awareness of the nutritional status of patients undergoing major elective surgery has lead to the finding that certain measurements of nutritional status either alone (dynamometry) or in combination (Prognostic Nutritional Index or the 'Sheffield' modified index) can identify patients at high risk from developing complications. This study presents a comparison of forearm muscle dynamometry versus the 'Sheffield' index in 70 patients undergoing major elective abdominal and urological surgery. Both tests successfully identified patients at risk of major post operative complications and death, but they failed to predict those patients who developed minor post operative complications. These data suggest that only major postoperative complications relate to the pre-operative nutritional status of patients undergoing major elective surgery. Identification of a patient as being at high risk of developing major postoperative complications by dynamometry or the 'Sheffield' index should alert the surgeon to the possible requirement for perioperative nutritional support.


Toxicity of isolated Lantana (Lantana camara L) constituents to male and female guinea pigs.
A toxin fraction was obtained from Lantana camara L (red variety) leaves by batch extraction and column chromatography on silica gel (60-120 mesh). The main constituents of the toxin preparation were lantadene A and lantadene B and it was devoid of reduced lantadene A. Oral administration (125 mg/kg bwt) of the toxin to male and female guinea pigs caused icterus and photosensitization within 48 hr. All the affected animals had hepatomegaly and significant increases in conjugated and unconjugated bilirubin in blood plasma. The intoxicated animals of either sex had marked increases in acid phosphatase activity which was inhibited 45.77% and 49.35% by 1 mM tartrate in male and female animals respectively. The corresponding inhibition of acid phosphatase activity in control male and female guinea pigs was 15.91% and 20.33% respectively.


Accumulation of cyclic ADP-ribose measured by a specific radioimmunoassay in differentiated human leukemic HL-60 cells with all-trans-retinoic acid.
Cyclic adenosine diphosphoribose (cADPR) is a novel candidate for the mediator of Ca2+ release from intracellular Ca2+ stores. The formation of this cyclic nucleotide is catalyzed by not only Aplysia ADP-ribosyl cyclase but also an ecto-form enzyme of NAD+ glycohydrolase (NADase), which was previously identified as all-trans-retinoic acid (RA)-inducible CD38 in human leukemic HL-60 cells. In the present study, we developed a radioimmunoassay specific for cADPR, by which more than 100 fmol of cADPR could be detected without any interference by other nucleotides. The possible involvement of CD38 in the formation of cellular cADPR was investigated with the radioimmunoassay method. A marked increase in cellular cADPR was accompanied by all-trans-RA-induced differentiation of HL-60 cells. Moreover, a high level of cellular cADPR was observed in other leukemic cell lines, in which CD38 mRNA was expressed. Thus, CD38, which was initially identified as an NADase, appeared to be responsible for the formation of cellular cADPR.


Notch2 regulates matrix metallopeptidase 9 via PI3K/AKT signaling in human gastric carcinoma cell MKN-45.
To clarify the role of activated Notch2 in the invasiveness of gastric cancer. To investigate the invasiveness of silencing Notch2 gene expression, we established a Notch2 small interfering RNA (siRNA) transfected cell line using the MKN-45 gastric cancer cell line. After the successful transfection confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, migration and invasion assays were employed to evaluate the aggressiveness of the gastric cancer. RT-PCR and Western blottings were employed to confirm the down-regulation of Notch2 and to evaluate the expression of epithelial mesenchymal transition-related gene matrix metallopeptidase 9 (MMP9), Akt, p-Akt. To confirm the relationship between PI3K-Akt and MMP9, the PI3K inhibitor LY294002 was used to treat MKN-45 cells. Notch2 expression was dramatically decreased after Notch2 siRNA transfection (100.00% ± 9.74% vs 11.61% ± 3.85%, P < 0.01 by qRT-PCR). There was also a marked reduction of Notch target gene Hes1 (100.00% ± 4.74% vs 61.61% ± 3.58%, P < 0.05) at the mRNA, indicating an inhibition of Notch signaling. Inhibition of Notch signaling was also confirmed by the marked reduction of Notch2 intracellular domain at the protein levels (100.00% ± 9.74% vs 65.61% ± 7.58%, P < 0.05). Down-regulation of Notch2 by siRNA enhanced tumor cell invasion (100.00% ± 21.64% vs 162.22% ± 16.84%, P < 0.05) and expression of MMP9 (1.56 fold, P < 0.05), and activated the pro-MMP9 protein to its active form (1.48 fold, P < 0.05). There was no significant difference in the protein levels of Akt between the two groups (100.00% ± 10.87% vs 96.61% ± 7.33%, P > 0.05), while down-regulation of Notch2 elevated p-Akt expression (100.00% ± 9.87% vs 154.61% ± 13.10%, P < 0.05). Furthermore, p-Akt and MMP9 was down-regulated in response to the inhibitor LY294002 (p-Akt 100.00% ± 8.87% vs 58.27% ± 5.01%, P < 0.05; MMP9 100.00% ± 9.17% vs 50.03% ± 4.88%, P < 0.05). Notch2 may negatively regulate cell invasion by inhibiting the PI3K-Akt signaling pathway in gastric cancer.


RNA interference targeting mutant p53 inhibits growth and induces apoptosis in DU145 human prostate cancer cells.
p53 is the most frequently mutated tumor suppressor gene in human cancer. Recent studies have indicated that p53 mutants not only lose tumor suppression activity but also gain novel oncogenic functions that contribute to tumor malignancy. In this study, we explored mutant p53 as a target for novel anti-cancer treatment in prostate cancer. Using the DU145 human androgen-independent prostate cancer cell line, we show that silencing of mutant p53 gene by RNA interference led to significant inhibition of cell viability and growth, which was associated with cell cycle arrest at G1 and G2/M phase, and ultimately induced massive apoptosis. Mechanistically, p53-siRNA inhibited phosphatidylinositol 3'-kinase/Akt signaling pathway, which might be responsible for the reduced proliferation and apoptosis induction. These findings suggest that RNA interference targeting mutant p53 may serve as a novel therapeutic strategy for the treatment of androgen-independent prostate cancer.


Computerized tomography of the patellofemoral joint before and after lateral release or realignment.
Computerized tomography (CT) provides an accurate diagnostic tool for evaluating tilt and subluxation of the patella. Using a previously described technique, this study reviews 62 computerized tomograms, including those of 21 patients before and after lateral release or anteromedial tibial tubercle transfer. One patient was evaluated before and after soft-tissue realignment of the patella. Additional CT studies were evaluated to establish the most appropriate reference line for determining patellar tilt. Results show that lateral retinacular release effectively reduces abnormal patellar tilting as determined by CT. Anteromedial tibial tubercle transfer similarly reduces abnormal tilt. Patellar subluxation may improve substantially following either lateral release or anteromedial tibial tubercle transfer, but this study suggests that correction of subluxation is less consistent than reduction of abnormal tilt with tibial tubercle transfer or lateral release alone. Once articular degeneration has progressed to the point of facet collapse, lateral release fails to restore normal tilt.


NF-κB-regulated VentX expression mediates tumoricidal effects of chemotherapeutics at noncytotoxic concentrations.
Limited therapeutic efficacy and severe side effects represent the central hurdles facing cancer chemotherapy. Immune suppression within tumor immune microenvironments (TIME) has been implicated in chemoresistance. In this study, using a TIME-enabling model system (TIME-EMS), we demonstrate that the chemotherapeutic agent doxorubicin has cytocidal effects on tumor cells at high dosage but induces changes in the immune landscape of the TIME at low noncytotoxic concentrations via NF-κB-mediated induction of homeobox protein VentX expression in tumor-associated macrophages (TAMs). We demonstrated that VentX-regulated TAMs drastically promote tumor chemosensitivity >10-fold but exert little effect on chemotoxicity to normal cells through activating cytotoxic T lymphocytes in a tumor-specific manner. Supported by the in vivo synergy of VentX-regulated TAMs and low-dosage noncytotoxic doxorubicin, our data suggest a cell-death-independent immune mechanism for improving the therapeutic index of chemotherapeutic agents.


[A polarographic method in assessing the cultural properties of the EV vaccinal strain].
The kinetic and regulatory characteristics of the oxidation metabolism of glucose, whose differences in collection cultures correspond to their capacity for growth in fluid culture medium under the conditions of aeration and the periodic addition of glucose, have been proposed on the basis of the polarographic method used for the registration of microbial breath in vaccine strain EV. Pronounced metabolic disturbances resulting from the prolonged storage of the stock culture and working standards of strain EV lead to the deterioration of their working properties.


Synthesis, crystal structure and photoluminescence of a cyclometalated Iridium(III) coumarin complex.
In this paper a new cyclometalated iridium(III) coumarin complex, Ir(III)bis(3-(2-benzothiazolyl)coumarinato N,C(4))(acetylacetonate) (Ir(L)2(acac)), was synthesized and characterized. X-ray crystallography demonstrated that the iridium(III) ion is hexacoordinated by two C atoms and two N atoms from 3-(2-benzothiazolyl)coumarinato ligands and two O atoms from acac ligand, displaying distorted octahedral coordination geometry. The Ir(L)2(acac) complex has good thermal stability with less than 2 % weight-reduction occurring at 300 °C, and exhibits strong reddish orange emission. The results shown that Ir(L)2(acac) is useful for fabrication organic light-emitting diodes.


Blood leukocyte responses to extracorporeal circulation. I. Short term extracorporeal circulation in dogs without and with extracorporeal irradiation.
Short term (1 h) extracorporeal circulation without or with irradiation of blood was performed in two normal dogs in a series of experiments. The granulocyte count was constantly diminished, while the lymphocytes did not show any particular change in their concentration. In the majority of the experiments a decrease of the CFU-C content occurred to less than 70% of the initial level. There was no difference in the results of experiments with or without irradiation. In the "bag to bag" procedures, no significant change in the blood leukocyte counts including CFU-C, was established.


Multiple resistance to ACCase and AHAS-inhibiting herbicides in shortawn foxtail (Alopecurus aequalis Sobol.) from China.
Shortawn foxtail (Alopecurus aequalis) is a troublesome grass weed infesting winter wheat and oilseed rape productions in China. Fenoxaprop-p-ethyl and mesosulfuron-methyl failed to control shortawn foxtail of AHSX-1 population collected from a wheat field in Shou County, Anhui province. Molecular analyses revealed that Asp2078Gly mutation of ACCase and Trp574Leu mutation of AHAS were present in plants of the AHSX-1 population. The homozygous plants were isolated and cultured until seed maturity. Whole-plant herbicide bioassays were conducted in the greenhouse using the purified seeds of F1 generation. Dose-response experiments showed that the AHSX-1 population has evolved a very high level resistance to fenoxaprop-p-ethyl (RI = 275) and mesosulfuron-methyl (RI = 788). To determine the sensitivity to other herbicides, assays were conducted at the single recommended rate of each herbicide. Based on the results, the AHSX-1 population was considered to be highly resistant to clodinafop-propargyl, pyroxsulam and flucarbazone-sodium, moderately or highly resistant to quizalofop-p-ethyl, clethodim, sethoxydim and pinoxaden, and susceptible to isoproturon and chlorotoluron. This is the first report of Asp2078Gly mutation in shortawn foxtail and the two robust dCAPS markers designed could quickly detect Asp2078 and Trp574 mutations in ACCase and AHAS gene of shortawn foxtail, respectively.


Rice Hull-Derived Carbon for Supercapacitors: Towards Sustainable Silicon-Carbon Supercapacitors.
A simple and effective mixing carbonization-activation process was developed to prepare rice hull-derived porous Si-carbon materials. The morphologies and pore structures of the materials were controlled effectively without any loading or additions at various carbonization temperatures. The structures of the samples changed from large pores and thick walls after 800 ∘C carbonization to small pores and thin walls after 1000 ∘C carbonization. An additional alkali activation-carbonization process led to coral reef-like structures surrounded by squama in the sample that underwent 900 ∘C carbonization (Act-RH-900). This optimal material (Act-RH-900) had a large specific surface area (768 m2 g-1), relatively stable specific capacitance (150.8 F g-1), high energy density (31.9 Wh kg-1), and high-power density (309.2 w kg-1) at a current density of 0.5 A g-1 in 1 M KOH electrolyte, as well as a good rate performance and high stability (capacitance retention > 87.88% after 5000 cycles). The results indicated that Act-RH-900 is a promising candidate for capacitive applications. This work overcomes the restrictions imposed by the complex internal structure of biomass, implements a simple reaction environment, and broadens the potential applicability of biomass waste in the field of supercapacitors.


A sensitive and simple method for determination of IgM in cerebrospinal fluid by a solid-phase enzyme-immunoassay. Comparison of two different methods.
A sensitive and simple solid-phase enzyme-immunoassay for quantitative determination of IgM in cerebrospinal fluid has been developed. This method is based on a sandwich technique and is much more sensitive than the competition enzyme-immunoassay which we described previously. The IgM values obtained by this sandwich technique were well correlated with those by the competition technique. This new method is considered to be more suitable for the estimation of IgM in cerebrospinal fluid because of its simplicity and good sensitivity.


Fluorescent tracers as potential candidates for double labeling of descending brain neurons in larval lamprey.
In larval lamprey, seven fluorescent tracers were tested as potential candidates for retrograde double labeling of descending brain neurons: Fluoro Gold (FG); fluorescein dextran amine (FDA); True Blue (TB); cascade blue dextran amine (CBDA); Fast Blue (FB); Texas red dextran amine (TRDA); and tetramethylrhodamine dextran amine (RDA). The first tracer (FG, TB, FB, or CBDA) was applied to the spinal cord at 40% body length (BL). In separate experiments, the second tracer (TRDA or RDA) was applied to the spinal cord at 20% BL. The tracer combination FG/TRDA was found to have the best optical properties for double labeling. However, application of FG to the spinal cord with the method used for the other tracers resulted in labeling of 'lateral cells' along the sides of the rhombencephalon that were presumed to be non-neuronal and that obscured some of the descending brain neurons. Control experiments suggested that FG was transported in the circulation to the brain, where the tracer was taken up by lateral cells. Therefore, a special technique was developed for applying FG to the spinal cord without allowing the tracer to enter the circulation. In larval lamprey, this important double-labeling technique that was developed for TRDA and FG is being used to examine axonal regeneration and projection patterns of descending brain neurons.


Social inequalities in breast cancer screening: evaluating written communications with immigrant Haitian women in Montreal.
The province of Quebec (Canada) has implemented a breast cancer screening program to diagnose this cancer at an early stage. The strategy is to refer women 50 to 69 years old for a mammogram every two years by sending an invitation letter that acts as a prescription. Ninety per cent (90%) of deaths due to breast cancer occur in women aged 50 and over. Numerous studies have shown social inequalities in health for most diseases. With breast cancer, a significant paradox arises: its incidence is lower among disadvantaged women and yet, more of them die from this disease. The health care system might play a role in this inequality. The scientific literature documents the potential for creating such inequalities when prevention does not consider equity among social groups. Immigrant women are often disadvantaged. They die of breast cancer more than non-immigrants. Studies attribute this to late-stage diagnosis due to poor adherence to mammography screening programs. The main objective of our research is to assess how Haitian immigrant women in Montreal are reached by the Quebec Breast Cancer Screening Program, and specifically how they perceive the mammogram referral letter sent by the program. The study uses a two-step qualitative method: i) In-depth interviews with influential community workers to identify the most relevant issues; ii) Focus groups with disadvantaged women from Montreal's Haitian community. A mammogram referral letter from the Breast Cancer Screening Program may be a barrier to compliance with mammography by underprivileged Haitian women in Montreal. This might be attributable to a low level of literacy, poor knowledge of the disease, and lack of financial resources. Barriers may be underestimated in underprivileged immigrant and non-immigrant communities. A preventive strategy must be adapted to different sub-groups and must also take into account lower literacy levels. To increase mammography uptake, it is crucial that the benefits of prevention be clearly identified and described in understandable terms. Finally, economic access to follow-up measures should be considered.


USP25 regulates KEAP1-NRF2 anti-oxidation axis and its inactivation protects acetaminophen-induced liver injury in male mice.
Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor responsible for mounting an anti-oxidation gene expression program to counter oxidative stress. Under unstressed conditions, Kelch-like ECH-associated protein 1 (KEAP1), an adaptor protein for CUL3 E3 ubiquitin ligase, mediates NRF2 ubiquitination and degradation. We show here that the deubiquitinase USP25 directly binds to KEAP1 and prevents KEAP1's own ubiquitination and degradation. In the absence of Usp25 or if the DUB is inhibited, KEAP1 is downregulated and NRF2 is stabilized, allowing the cells to respond to oxidative stress more readily. In acetaminophen (APAP) overdose-induced oxidative liver damage in male mice, the inactivation of Usp25, either genetically or pharmacologically, greatly attenuates liver injury and reduces the mortality rates resulted from lethal doses of APAP.


Molecular variability of Hop stunt viroid: identification of a unique variant with a tandem 15-nucleotide repeat from naturally infected plum tree.
For this study, 68 plum samples were collected from 12 provinces of China. Low molecular weight RNAs were extracted and used for dot-blot, reverse transcription-polymerase chain reaction (RT-PCR), return-polyacrylamide gel electrophoresis, and biological indexing using cucumber. Results showed that 15 out of the 68 plum samples were positive for Hop stunt viroid (HSVd). Four positive samples were selected for cloning and sequence analysis. Results indicated that most HSVd sequences from the plum in China had 1-3 nucleotide changes from the closest HSVd in GenBank. In addition, the sample PB21 (cv. 'Friar') collected from Hebei province had one sequence with a 15-nt duplication (named HSVd D-15) at the 244/245 position in the lower central region. By biological indexing, cucumber seedlings, an indicator plant for HSVd, were inoculated with RNAs directly extracted from the original plum source (PB21, cv. 'Friar'). Nucleotide sequencing analysis of the progeny showed that HSVd, but not HSVd-D15, was recovered from inoculated cucumbers. Although very unlikely, the possibility that this extra sequence was the result of a PCR artifact cannot be completely ruled out.


Congenital Harlequin syndrome as an isolated phenomenon: A case report and review of the literature.
Harlequin syndrome (HS) is a rare autonomic disorder due to a hemifacial cutaneous sympathetic denervation. It is characterized by unilateral diminished sweating and flushing of the face even though after heat or prolonged exercise. It is typically acquired. Congenital cases only represent a 6% of all individuals with HS. All congenital HS cases reported so far showed a concomitant Horner syndrome, probably due to a stellate ganglion involvement. HS represents an uncommon autonomic disorder due to a hemifacial cutaneous sympathetic denervation. It is clinically characterized by a dramatic alteration in facial appearance: ipsilateral denervated pale and dry half from the other intact red and moist half. We present, to the best of our knowledge, the first case of a patient with a congenital HS as an isolated phenomenon.


Extracellular vesicles derived from Akkermansia muciniphila promote placentation and mitigate preeclampsia in a mouse model.
Preeclampsia (PE) is a multisystem disorder with high maternal morbidity and mortality rates. Currently, no practical therapeutic approach is available to prevent PE progression, except for early delivery. Gut dysbiosis is associated with PE development. Previous data showed that the abundance of Akkermansia muciniphila (Am) was lower in patients with PE than in normotensive pregnant women. Here, in this study, decreased abundance of Am was observed in a PE mouse model. Also, we found that administration with Am could significantly attenuate systolic blood pressure, promote foetal growth and improve the placental pathology in mice with PE. Moreover, Am-derived extracellular vesicles (AmEVs) were transferred from the gastrointestinal (GI) tract to the placenta and mitigated pre-eclamptic symptoms in PE mice. These beneficial effects of AmEVs were mediated by enhanced trophoblast invasion of the spiral artery (SpA) and SpA remodelling through activation of the epidermal growth factor receptor (EGFR)-phosphatidylinositol-3-kinase (PI3K)-protein kinase B (AKT) signalling pathway. Collectively, our findings revealed the potential benefit of using AmEVs for PE treatment and highlighted important host-microbiota interactions.


Monoterpene alcohols induced by sweet potato weevil larvae deter conspecific adults from feeding and oviposition.
Intraspecific competition is shared in the insect world, especially under the condition of limited food and space resources. To avoid intraspecific competition and increase offspring survival, insects have evolved various effective strategies. A widely-accepted tactic is employing chemical cues, which are frequently utilized as indicators of conspecific colonization. The sweet potato weevil (SPW), Cylas formicarius, is a destructive pest of sweet potatoes. Its larvae bore into sweet potatoes and alter the emission of odors. The present study aimed to investigate whether volatiles associated with SPW larvae feeding influence the behavioral preference of conspecific adults. Volatiles from SPW larvae-infested sweet potatoes were collected by a head-space method and analyzed using gas chromatography-electroantennogram detector (GC-EAD) and gas chromatography-mass spectrometry (GC-MS). Five compounds eliciting EAD responses from the antennae of both male and female adult SPW were identified from sweet potatoes with the third-instar larvae, including linalool, citronellol, nerol, geraniol, and ipomeamarone. Four monoterpene alcohols significantly repelled SPW adults from feeding and oviposition at higher doses in the behavioral preference bioassays. Among them, geraniol displayed the strongest repellent activities for SPW feeding and oviposition. These results suggested that SPW larvae could reduce colonization of adult SPWs by inducing monoterpene alcohols, thereby avoiding intraspecific competition. The present study demonstrated that volatile monoterpene alcohols induced by SPW larvae are chemical cues of larvae occupation for SPW adults to change their behavioral preference. Unveiling factors that mediate avoidance of intraspecific competition could help develop repellents or oviposition deterrents for SPW control. © 2023 Society of Chemical Industry.


Patterns of gene expression in pig adipose tissue: insulin-like growth factor system proteins, neuropeptide Y (NPY), NPY receptors, neurotrophic factors and other secreted factors.
Although cDNA microarray studies have examined gene expression in human and rodent adipose tissue, only one microarray study of adipose tissue from growing pigs has been reported. Total RNA was collected at slaughter from outer subcutaneous adipose tissue (OSQ) and middle subcutaneous adipose tissue (MSQ) from gilts at 90, 150, and 210 d (n=5 age(-1)). Dye labeled cDNA probes were hybridized to custom porcine microarrays (70-mer oligonucleotides). Gene expression of insulin-like growth factor binding proteins (IGFBPs), hormones, growth factors, neuropeptide Y (NPY) receptors (NPYRs) and other receptors in OSQ and MSQ changed little with age in growing pigs. Distinct patterns of relative gene expression were evident within NPYR and IGFBP family members in adipose tissue from growing pigs. Relative gene expression levels of NPY2R, NPY4R and angiopoietin 2 (ANG-2) distinguished OSQ and MSQ depots in growing pigs. We demonstrated, for the first time, the expression of IGFBP-7, IGFBP-5, NPY1R, NPY2R, NPY, connective tissue growth factor (CTGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) genes in pig adipose tissue with microarray and RT-PCR assays. Furthermore, adipose tissue CTGF gene expression was upregulated while NPY and NPY2R gene expression were significantly down regulated by age. These studies demonstrate that expression of neuropeptides and neurotrophic factors in pig adipose tissue may be involved in regulation of leptin secretion. Many other regulatory factors were not influenced by age in growing pigs but may be influenced by location or depot.


Future research directions to identify risks and mitigation strategies for neurostructural, ocular, and behavioral changes induced by human spaceflight: A NASA-ESA expert group consensus report.
A team of experts on the effects of the spaceflight environment on the brain and eye (SANS: Spaceflight-Associated Neuro-ocular Syndrome) was convened by NASA and ESA to (1) review spaceflight-associated structural and functional changes of the human brain and eye, and any interactions between the two; and (2) identify critical future research directions in this area to help characterize the risk and identify possible countermeasures and strategies to mitigate the spaceflight-induced brain and eye alterations. The experts identified 14 critical future research directions that would substantially advance our knowledge of the effects of spending prolonged periods of time in the spaceflight environment on SANS, as well as brain structure and function. They used a paired comparison approach to rank the relative importance of these 14 recommendations, which are discussed in detail in the main report and are summarized briefly below.


OSKM Induce Extraembryonic Endoderm Stem Cells in Parallel to Induced Pluripotent Stem Cells.
The reprogramming factors OCT4, SOX2, KLF4, and MYC (OSKM) can reactivate the pluripotency network in terminally differentiated cells, but also regulate expression of non-pluripotency genes in other contexts, such as the mouse primitive endoderm. The primitive endoderm is an extraembryonic lineage established in parallel to the pluripotent epiblast in the blastocyst, and is the progenitor pool for extraembryonic endoderm stem (XEN) cells. We show that OSKM induce expression of endodermal genes, leading to formation of induced XEN (iXEN) cells, which possess key properties of blastocyst-derived XEN cells, including morphology, transcription profile, self-renewal, and multipotency. Our data show that iXEN cells arise in parallel to induced pluripotent stem cells, indicating that OSKM drive cells to two distinct cell fates during reprogramming.


f-MOPSO/Div: an improved extreme-point-based multi-objective PSO algorithm applied to a socio-economic-environmental conjunctive water use problem.
In this paper, a new version of the multi-objective particle swarm optimizer named the Diversity-enhanced fuzzy multi-objective particle swarm optimization (f-MOPSO/Div) algorithm is proposed. This algorithm is an improved version of our recently proposed f-MOPSO. In the proposed algorithm, a new characteristic of the particles in the objective space, which we named the "extremity," is also evaluated, along with the Pareto dominance, to appoint proper guides for the particles in the search space. Three improvements are applied to the f-MOPSO to mitigate its shortcomings, generating f-MOPSO/Div: (1) selecting the global best solution based on the diversity of the extreme solutions, (2) impeding the particles to be trapped in the local optima using a mutation scheme based on the dynamic probability, and (3) removing the pre-optimization process. To validate f-MOPSO/Div, it was compared with some other popular multi-objective algorithms on 14 standard low- and high-dimensional test problem suites. After the comparative results indicated the outperformance of the proposal, the f-MOPSO/Div was applied to solve an optimal conjunctive water use management problem, in a semi-arid study area in west-central Iran, over a 13-year long-term planning period with two main objectives: (1) maximizing the aquifer sustainability as an environmental goal, and (2) maximizing the crop yields as a socio-economic goal. As the results suggest, the cumulative groundwater level drawdown is considerably decreased over the whole planning period to make the aquifer sustainable, while the water productivity is held at a desirable level, demonstrating the superiority of the f-MOPSO/Div when also applied to solve a large-scale real-world optimization problem.


Ecotoxicological implications of leachates from concrete demolition debris on oligochaetes: survival and oxidative stress status.
Urbanization and population growth demand the construction of structures to facilitate the need for space, and old infrastructures must make space for new ones leading to demolition and concrete debris. In addition to demolition, aging and weather are factors leading to concrete deterioration and, thus, a new challenge as an environmental pollutant. Studies on how concrete debris and leachate affect biota in the environment are limited. The present study aimed to understand the effects of leachate from various sizes of concrete debris on the three oligochaete species Enchytraeus crypticus, Tubifex, and Lumbriculus variegatus. Acute toxicity testing was carried out to determine the adverse effects over time. The oligochaetes' survival was monitored as well as the activity of the biotransformation enzyme glutathione S-transferase and the antioxidative enzyme catalase as indicators of the oxidative stress status. Leachate from the smallest concrete particle size (<1 mm) was found to be the most toxic as it caused, on average, 6-fold increased oligochaete mortality compared to the larger pieces (2-5 cm) after 96 h of exposure, potentially due to the larger surface area facilitating the release of toxicants. Substrate buffered the toxic effect of the leachate with 42 ± 12% fewer mortalities and reduced adverse effects on the enzymes. Of the three oligochaetes, E. crypticus was the most resilient to the concrete leachate. The study is the first to investigate the effects of concrete leachate on oligochaetes.