Paternal Combined Botanicals Contribute to the Prevention of Estrogen Receptor-Negative Mammary Cancer in Transgenic Mice.
Parental nutritional interventions have considerably affected gametogenesis and embryogenesis, leading to the differential susceptibility of offspring to chronic diseases such as cancer. Moreover, combinatorial bioactive diets are more efficacious in ameliorating epigenetic aberrations in tumorigenesis. We sought to investigate the transgenerational influence and epigenetic regulation of paternal sulforaphane (SFN)-rich broccoli sprouts (BSp) and epigallocatechin-3-gallate (EGCG)-rich green tea polyphenols (GTPs) consumption in the prevention of estrogen receptor-negative [ER(-)] mammary cancer in transgenic mice. Human breast cancer cells were used to detect cell viability and epigenetic-related gene expression after treatment with EGCG and/or SFN. Twenty-four C3 or HER2/neu males were randomly assigned into 4 groups and treated with control, 26% BSp (w/w) in food, 0.5% GTPs (w/v) in drinking water or combined BSp and GTPs for 7 wk before mating. Tumor growth of nontreated female pups was monitored weekly for 19 wk (C3) and 25 wk (HER2/neu). Tumor- and epigenetic-related protein expression and enzyme activities in mammary tumors were measured. Sperms were isolated from treated males for RNA sequencing and reduced-representation bisulfite sequencing analysis. Data were analyzed with a 2-factor or 3-factor analysis of variance. EGCG and SFN inhibited breast cancer cell growth via epigenetic regulation. Combined BSp and GTPs synergistically (combination index < 1) suppressed tumor growth over time (P < 0.001) in 2 mouse models. Key tumor-related proteins were found differentially expressed (P < 0.05) along with epigenetic regulations in offspring mammary tumors. The transcriptome profile of sperm derived from dietary-treated males revealed differentially expressed genes correlated with spermatogenesis and breast cancer progression. DNA methylomes of the sperm and further integrated analysis with transcriptomes indicate that DNA methylation alone may not contribute to sufficient regulation in dietary-treated sperm pronucleus, leading to offspring tumor suppression. Collectively, paternal consumption of combined BSp and GTPs shows potential for preventing ER(-) mammary cancer through transgenerational effects. J Nutr 2023;xx:xx-xx.


Development of IC-ELISA for detection of organophosphorus pesticides in water.
Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC50 of 0.428 micro g/mL with a detection limit of 0.0125 micro g/mL to parathion. The assay also indicated that the IC50 values of pirimiphos and dichlorvos were 0.331 micro g/mL and 1.25 micro g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 micro g/mL and 0.048 micro g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.


The comparative short-term effectiveness of iron dosing and formulations in US hemodialysis patients.
Intravenous iron is used widely in hemodialysis, yet there are limited data on the effectiveness of contemporary dosing strategies or formulation type. We conducted a retrospective cohort study using data from the clinical database of a large dialysis provider (years 2004-2008) merged with administrative data from the US Renal Data System to compare the effects of intravenous iron use on anemia management. Dosing comparisons were bolus (consecutive doses ≥100 mg exceeding 600 mg during 1 month) versus maintenance (all other iron doses during the month); and high (>200 mg over 1 month) versus low dose (≤200 mg over 1 month). Formulation comparison was administration of ferric gluconate versus iron sucrose over 1 month. Outcomes were hemoglobin, epoetin dose, transferrin saturation, and serum ferritin during 6 weeks of follow-up. We identified 117,050 patients for the dosing comparison, and 66,207 patients for the formulation comparison. Bolus dosing was associated with higher average adjusted hemoglobin (+0.23 g/dL; 95% confidence interval [CI], 0.21-0.26), transferrin saturation (+3.31%; 95% CI, 2.99-3.63), serum ferritin (+151 μg/L; 95% CI, 134.9-168.7), and lower average epoetin dose (-464 units; 95% CI, -583 to -343) compared with maintenance. Similar trends were observed with high-dose iron versus low-dose. Iron sucrose was associated with higher adjusted average hemoglobin (+0.16 g/dL; 95% CI, 0.12-0.19) versus ferric gluconate. Strategies favoring large doses of intravenous iron or iron sucrose lead to improved measures of anemia management. These potential benefits should be weighed against risks, which currently remain incompletely characterized.


Effects of ultrasound-assisted low-concentration chlorine washing on ready-to-eat winter jujube (Zizyphus jujuba Mill. cv. Dongzao): Cross-contamination prevention, decontamination efficacy, and fruit quality.
Wash water is circulated for use in the minimal processing industry, and inefficient disinfection methods can lead to pathogen cross-contamination. Moreover, few disinfection strategies are available for ready-to-eat fruits that do not need to be cut. In this study, the use of chlorine and ultrasound, two low-cost disinfection methods, were evaluated to disinfect winter jujube, a delicious, nutritious, and widely sold fruit in China. Ultrasound treatment (28 kHz) alone could not decrease the cross-contamination incidence of Escherichia coli O157:H7, non-O157 E. coli, and Salmonella Typhimurium, and free chlorine treatment at 10 ppm decreased the incidence from 55.00% to 5.00% for E. coli O157:H7, 65.00% to 6.67% for non-157 E. coli, and 70.00% to 6.67% for S. Typhimurium. The cross-contamination incidence was completely reduced (pathogens were not detected in sample) when the treatments were combined. The counts of aerobic mesophiles, aerobic psychrophiles, molds, yeasts, and three pathogens in the group subjected to combination treatment (28 kHz ultrasound + 10 ppm free chlorine) were significantly lower than those in the control, chlorine-treated, and ultrasound-treated groups during storage (0-7 d at 4 °C). Analysis of weight loss, sensory quality (crispness, color, and flavor), instrument color (a*/b*), soluble matter contents (total soluble solids, reducing sugar, total soluble sugar, and titratable acid), and nutritional properties (ascorbic acid and polyphenolic contents) indicated that treatment with ultrasound, chlorine, and their combination did not lead to additional quality loss compared with properties of the control. Additionally, the activities of phenylalanine ammonia-lyase and polyphenol oxidase were not significantly increased in the treatment group, consistent with the quality analysis results. These findings provide insights into disinfection of uncut ready-to-eat fruits using a minimum dose of disinfectant for cross-contamination prevention under ultrasonication. The use of ultrasound alone to decontaminate fresh produce is accompanied by a high risk of pathogen contamination, and the use of sanitizers to decrease cross-contamination incidence is recommended.


Studies on lithium transport across the red cell membrane. VI. Properties of a sulfhydryl group involved in ouabain-resistant Na+-Li+ (and Na+-Na+) exchange in human and bovine erythrocytes.
The reactivity of the SH-group essential for ouabain-resistant Na+-Li+ (and Na+-Na+) exchange and its location within the membrane are studied on human and beef erythrocytes and beef red cell ghosts. N-ethylmaleimide (NEM), 1,6-hexane dimaleimide, and iodoacetamide can induce an irreversible, partial inhibition of Na+-Li+ exchange in erythrocytes of the two species. The development of the inhibition due to the alkylating agents is greatly accelerated by external Na+ and Li+. The inhibition takes 3 min (NEM) and 60 min (iodoacetamide) to come to completion in isotonic Na+ media, but is hardly detectable in choline+, K+ or Mg2+ media. The transport site of the exchange system and the site promoting NEM binding exhibit similar affinities for external Na+. The impermeable, monofunctional glutathione derivative of 1,6-hexane dimaleimide does not inhibit Na+-Li+ exchange. The mercurials PCMBS, PCMB, and Hg2+ inhibit Na+-Li+ exchange in beef, but not in human erythrocytes. The inhibitory action of PCMBS, being slightly accelerated by external Na+, is fully reversed by penetrating thiols such as 2-mercaptoethanol, whilst glutathione, an impermeable thiol, is ineffective. Pretreatment with PCMBS affords partial protection from the irreversible inhibition caused by NEM. Oxidation with copper orthophenanthroline inhibits Na+-Li+ exchange only when performed in the presence of penetrating thiols such as 2-mercaptoethanol. It is concluded that the SH-reagents studied inhibit Na+-Li+ exchange by modifying an essential SH-group of a membrane protein in such a way that the turnover number of the exchange system is reduced. This SH-group is separated from both the red cell exterior and interior by a penetration barrier and seems to be distinct from the cation binding site. The action of external Na+ and Li+ in promoting the reaction of alkylating inhibitors is interpreted to result from a conformational change of the transport protein induced by the binding of external Na+ or Li+.


Introduction to Danish (nationwide) registers on health and social issues: structure, access, legislation, and archiving.
Danish registers contain information on many important health and social issues. Because all Danish citizens have a unique personal identification number, linkage at the individual level between these nationwide registers and other data sources is possible and feasible. In this paper we briefly introduce selected Danish registers and the data structure and requirements forgetting access to data at Statistics Denmark, which is the main provider of register data. We introduce the Danish Data Archive and briefly present the Act on Processing of Personal Data, which is the legal foundation for analyses of register-based data in Denmark.


Myocardial Dysfunction in Cirrhotic Cardiomyopathy is Associated with Alterations of Phospholamban Phosphorylation and IL-6 Levels.
Decreased cardiac contractility has been observed in cirrhosis, but the mechanisms that initiate and maintain cardiac dysfunction are not entirely understood. We test the hypothesis that cirrhotic cardiomyopathy is related to deterioration of myocardial contractility due to alterations in calcium-handling proteins expression. In addition, we evaluated whether cardiac pro-inflammatory cytokine levels are associated with this process. Cirrhosis was induced by thioacetamide (TAA, 100 mg/kg/i.p., twice weekly for eight weeks). The myocardial performance was evaluated in isolated left ventricle papillary muscles under basal conditions and after inotropic challenge. The cardiac calcium handling protein expression was detected by Western blotting. Cardiac TNF-α and IL-6 levels were measured by ELISA. Thioacetamide induced liver cirrhosis, which was associated with cirrhotic cardiomyopathy characterized by in vivo left ventricular diastolic and systolic dysfunction as well as cardiac hypertrophy. In vitro baseline myocardial contractility was lower in cirrhosis. Also, myocardial responsiveness to post-rest contraction stimulus was declined. Protein expression for RYR2, SERCA2, NCX, pPBL Ser16 and L-type calcium channel was quantitatively unchanged; however, pPBL Thr17 was significantly lower while IL-6 was higher. Our study demonstrates that cirrhotic cardiomyopathy is associated with decreased cardiac contractility with alteration of phospholamban phosphorylation in association with higher cardiac pro-inflammatory IL-6 levels. These findings provided molecular and functional insights about the effects of liver cirrhosis on cardiac function.


Respiratory and allergic health effects in children living near agriculture: A review.
Exposure to pesticides and agricultural burning are likely to co-occur in agricultural communities, but these exposures have remained distinct bodies of research. We reviewed epidemiological studies to identify the respiratory health effects of children exposed to pesticides and agricultural burning through a systematic evaluation of peer-reviewed publications of children living in industrial agricultural areas. Two academic search databases (PubMed and Scopus) were queried for all available studies published in English before May 31st, 2021. The initial search combining both exposure metrics (pesticides and agricultural burning) yielded zero publications and thus the queries were performed and presented separately. Studies were categorized based on main exposure of interest (i.e., pesticides or agricultural burning) and by respiratory health outcome assessment (i.e., self-reported asthma, acute respiratory symptoms, and lung function measurements). In total we identified 25 studies that focused on pesticide exposures and children's respiratory health, and 12 studies that focused on exposure to agricultural burning and children's respiratory health. A majority of the pesticide studies (18/25) reported a positive association between exposure to pesticides and adverse childhood respiratory health effects. Similarly, most (11/12) of the agricultural burning studies also reported a positive association between exposure to agricultural burning and adverse respiratory health effects. The most frequently studied health outcomes in these publications were acute respiratory symptoms (n = 11 pesticides, n = 3 agricultural burning), followed by asthma (n = 9 pesticides, n = 3 agricultural burning), and lung function measurements (n = 5 pesticides, n = 6 agricultural burning). Although health outcome assessment differed between pesticide studies and agricultural burning studies, similar adverse respiratory health effects were observed across the majority of studies.


Performance of chemically synthesized Mn3O4/rGO nanocomposite for electrochemical supercapacitor: a cost-effective high-performance electrode.
The manganese oxide graphene oxide (Mn3O4/rGO) composite heterojunction with copper oxide is useful for the production of an electrochemical supercapacitor. The graphene oxide and manganese oxide composite have been synthesized by adopting a method of co-precipitation. The composite of Mn3O4/rGO was synthesized with different concentrations of Mn3O4 and rGO. The structural, morphological, electrochemical and supercapacitive properties of Mn3O4/rGO composite have been examined. The electrochemical and supercapacitive properties have been studied with regard to different substrates. The Mn3O4/rGO composite was deposited on different substrates such as steel, copper and brass. The CuO/Mn3O4/rGO shows relatively better specific capacitance (856 F g-1) and better stability (82% retention after 2000 cycles) than other substrates used. The present work describes the development of cost-effective and high-performance CuO/Mn3O4/rGO-based nanomaterials for supercapacitors. The CuO/Mn3O4/rGO composite can be used as a flexible supercapacitor device.


Degradation of gas-liquid gliding arc discharge on Acid Orange II.
The effects of pH value, initial concentration of dye solution and temperature on the degradation efficiency of Acid Orange II (AO7) using gas-liquid gliding arc discharge were investigated. The influences of pH value and temperature on degradation efficiency were not apparent. Increasing initial solution concentration caused the decrease of degradation rate and the increase of absolute degradation quantity. Considering energy efficiency and absolute degradation quantity, the gas-liquid gliding arc discharge is fit for treating high concentration organic wastewater. A possible mineralization pathway was proposed through the analysis of intermediate products detected by gas chromatograph coupled with mass spectrophotometer (GC-MS) and ion chromatograph (IC). Hydroxyl radicals reacted with the azo linkage-bearing carbon of a hydroxy-substituted ring, leading to the cleavage of -C-N- and degradation of AO7. The solution biodegradability was significantly improved (BOD(5)/COD from 0.02 to 0.43). The toxicity of intermediate products was lower than that of the initial Acid Orange II.


Functional characterization of three naturally occurring single nucleotide polymorphisms in the CES2 gene encoding carboxylesterase 2 (HCE-2).
Twelve single nucleotide polymorphisms (SNPs) in the human CES2 gene, which encodes a carboxylesterase, hCE-2 [human carboxylesterase 2 (EC 22.214.171.124)], have been reported in the Japanese. In this report, we have examined functional alterations of three SNPs, a nonsynonymous SNP (100C>T, R34W), an SNP at the splice acceptor site in intron 8 (IVS8-2A>G), and one newly discovered nonsynonymous SNP (424G>A, V142M). For the two nonsynonymous SNPs, the corresponding variant cDNAs were expressed in COS-1 cells. Both the R34W and V142M variants showed little esterase activities toward the anticancer agent irinotecan and two typical carboxylesterase substrates, p-nitrophenol acetate and 4-methylumbelliferyl acetate, although increased levels of cDNA-mediated protein expression were observed by Western blotting as compared with the wild type. To investigate a possible splicing aberration in IVS8-2A>G, an in vitro splicing assay was utilized and transcripts derived from CES2 gene fragments of the wild type and IVS8-2A>G were compared. Sequence analysis of the cloned transcripts revealed that IVS8-2A>G yielded mostly aberrantly spliced transcripts, including a deleted exon or a 32-bp deletion proximal to the 5' end of exon 9, which resulted in truncated hCE-2 proteins. These results suggested that 100C>T (R34W), 424G>A (V142M), and IVS8-2A>G are functionally deficient SNPs.


Ozone uptake by adult urban trees based on sap flow measurement.
The O(3) uptake in 17 adult trees of six urban species was evaluated by the sap flow-based approach under free atmospheric conditions. The results showed very large species differences in ground area scaled whole-tree ozone uptake (F(o)₃), with estimates ranging from 0.61 ± 0.07 nmol m(-2) s(-1) in Robinia pseudoacacia to 4.80 ± 1.04 nmol m(-2) s(-1) in Magnolia liliiflora. However, average F(o)₃by deciduous foliages was not significantly higher than that by evergreen ones (3.13 vs 2.21 nmol m(-2) s(-1), p = 0.160). Species of high canopy conductance for O(3) (G(o)₃) took up more O(3) than those of low G(o)₃, but that their sensitivity to vapour pressure deficit (D) were also higher, and their F(o)₃decreased faster with increasing D, regardless of species. The responses of F(o)₃to D and total radiation led to the relative high flux of O(3) uptake, indicating high ozone risk for urban tree species.


Contrasting long term temporal trends in perfluoroalkyl substances (PFAS) in eggs of the northern gannet (Morus bassanus) from two UK colonies.
We compared long-term (1977 to 2014) trends in concentrations of PFAS in eggs of the marine sentinel species, the Northern gannet (Morus bassanus), from the Irish Sea (Ailsa Craig) and the North Sea (Bass Rock). Concentrations of eight perfluorinated carboxylic acids (PFCAs) and three perfluorinated sulfonates (PFSAs) were determined and we report the first dataset on PFAS in UK seabirds before and after the PFOS ban. There were no significant differences in ∑PFAS or ∑PFSAs between both colonies. The ∑PFSAs dominated the PFAS profile (>80%); PFOS accounted for the majority of the PFSAs (98-99%). In contrast, ∑PFCAs concentrations were slightly but significantly higher in eggs from Ailsa Craig than in those from Bass Rock. The most abundant PFCAs were perfluorotridecanoate (PFTriDA) and perfluoroundecanoate (PFUnA) which, together with PFOA, comprised around 90% of the ∑PFCAs. The ∑PFSAs and ∑PFCAs had very different temporal trends. ∑PFSAs concentrations in eggs from both colonies increased significantly in the earlier part of the study but later declined significantly, demonstrating the effectiveness of the phasing out of PFOS production in the 2000s. In contrast, ∑PFCAs concentrations in eggs were constant and low in the 1970s and 1980s, suggesting minimal environmental contamination, but residues subsequently increased significantly in both colonies until the end of the study. This increase appeared driven by rises in long chain compounds, namely the odd chain numbered PFTriDA and PFUnA. PFOA, had a very different temporal trend from the other dominant acids, with an earlier rise in concentrations followed by a decline in the last 15 years in Ailsa Craig; later temporal trends in Bass Rock eggs were unclear. Although eggs from both colonies contained relatively low concentrations of PFAS, the majority had PFOS residues that exceeded a suggested Predicted No Effect Concentration and ~ 10% of the eggs exceeded a suggested Lowest-Observable-Adverse-Effect.


The homeostasis of iron and suppression of HO-1 involved in the protective effects of nimodipine on neurodegeneration induced by aluminum overloading in mice.
Aluminum intoxication can cause damage to the cognitive function and neurodegenerative diseases. In the present study, we investigated the role of iron homeostasis and heme oxygenase-1 (HO-1) expression in the protective effects of nimodipine on the neurodegeneration induced by aluminum overloading in mice. 2 microl of 0.25% aluminum chloride solution was intracerebroventricularly injected once a day for five days to induce the neurodegeneration of mice. Nimodipine was administered by intragastric gavage (80 mg/kg per day) for 30 days. We observed that nimodipine could improve the performance of behavior test related to the learning and memory function and ameliorate pathological changes of hippocampi caused by aluminum. Results of western blot, immunohistochemistry study, biochemical test and inductively coupled plasma-atomic emission spectrometry showed that nimodipine could suppress the increased expression of HO-1 protein, and decrease the elevation of both HO activity and iron level in hippocampi, induced by aluminum overloading. These results indicate that nimodipine can suppress the neurodegenerative development induced by aluminum overloading and the mechanism of its action is at least partly related to keeping the homeostasis of iron through blunting the expression of HO-1 in hippocampus.


Analysis of the antigen-specific IFN-gamma producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis.
Three 10 months old cattle were infected by the intratracheal route with 10(6)cfu of a field strain of Mycobacterium bovis. Blood samples were regularly collected for in vitro IFN-gamma production after antigenic stimulation. Peripheral blood cells of infected animals produced IFN-gamma in response to crude M. bovis antigens (live and heat-inactivated BCG and protein-purified derivative (PPD)) 3-4 weeks after infection. The ratio of the response to bovine PPD versus avian PPD indicated a specific sensitisation for M. bovis antigens. Three months post-infection (PI), animals were culled and M. bovis was cultured from tubercle lesions. At different time points, the frequency of specific M. bovis IFN-gamma producing CD4+, CD8+ and WC1+ T-cells in the peripheral blood was examined by flow cytometry. Two colour immunofluorescence staining of intracellular IFN-gamma and bovine cell surface molecules showed that both CD4+ and CD8+, but not WC1+, T-cells produced IFN-gamma following stimulation with PPD, live or killed BCG. In two animals analysed, the relative percentage of circulating IFN-gamma producing CD8+ cells decreased between week 5 and week 9 PI. The same evolution was not observed for IFN-gamma secreting CD4+ cells. Magnetic positive selection of T-cells from infected animals showed that CD4+ T-cells produced specific IFN-gamma only in the presence of antigen presenting cells (APCs). Positively selected CD8+ T-cells secreted IFN-gamma only in the presence of recombinant human IL-2 and APCs. In vitro depletion of the CD4+ T-cells, but not the depletion of CD8+ or WC1+ T-cells, resulted in abrogation of the specific IFN-gamma production showing the key role of this cell population for the specific IFN-gamma production.


Isolating the interface of an emulsion using X-ray scattering and tensiometry to understand protein-modulated alkylglyceride crystallisation.
Dairy proteins and mono- and diglycerides (MDG) are often used in unison to tailor the properties of dairy-based emulsions. However, there are significant gaps in our understanding of how proteins affect lipid crystallisation at the oil-water interface. We have used a unique combination of interfacially-sensitive techniques to elucidate the impact of dairy proteins on interfacial MDG crystal formation. The formation temperature of interfacial MDG crystals was assessed through interfacial tension studies via drop shape analysis. Small and Wide-Angle X-ray Scattering measurements were performed on isolated oil-water interfaces, allowing for in-situ interrogation of MDG crystal structure and concentration at and near the interface. Dairy proteins are seen to reduce the temperature at which MDG crystals form at the oil-water interface. The displacement of proteins upon interfacial crystal formation was also clearly observed in interfacial tension measurements. For the first time, lipid crystals formed at the oil-water interface have been characterised using X-ray scattering. All scattering studies showed no change to the MDG crystal structures at the oil-water interface in the presence of adsorbed proteins. The results demonstrate that informed selection of emulsifier components is critical to controlling interfacial crystallisation with concomitant impact on emulsion stability.


A prospective, randomized pilot trial of model-based warfarin dose initiation using CYP2C9 genotype and clinical data.
Rapid genetic screening for cytochrome P450 (CYP) 2C9 variants may play a role in improving the efficacy and safety of warfarin in individuals with CYP2C9 variants. The feasibility of prospective CYP2C9 model-based warfarin dosing has not yet been assessed. To evaluate the feasibility of applying a CYP2C9 gene-based warfarin dosing model in clinical practice. Prospective, randomized, single-blinded clinical pilot trial. Large multispecialty group practice. Candidates were recruited from a list of clinic patients eligible for warfarin initiation. This included patients with newly diagnosed thromboembolic disease or atrial arrhythmia, as well as patients anticipating elective valvuloplasty or arthroplasty. Patients who previously received warfarin were excluded. Subjects were randomized to receive either 1) a standard initiation dose of 5 mg warfarin/day, or 2) rapid CYP2C9 genotyping and an initiation dose determined using parameters estimated from a previously published multivariate model [including age, body size, co-morbidity (e.g., diabetes), clinical indication (e.g., valvuloplasty) and CYP2C9 genotype]. Primary outcome measurements were patient willingness to participate, physician willingness to refer, sample processing time, ability to administer calculated dosage and adequacy of follow-up. This pilot trial was designed to assess the feasibility of model-based warfarin dosing. Power was insufficient for statistical comparison of adverse event rates. Forty-three of 117 patients had no prior warfarin treatment and were eligible. Five declined to participate. Twenty patients were randomized to a standard initiation dose of 5 mg daily. Eighteen patients were randomized to model-based dosing. All but one participant received the assigned initiation dose. Blood draw to dosage calculation time (including genotyping) required approximately 4 hours. Six adverse events occurred within the standard dosing group, and two adverse events occurred within the model-based dosing group. Prospective application of a multivariate CYP2C9 gene-based warfarin dosing model is feasible.


Right-sided varicocele associated with situs inversus.
In the overwhelming majority of cases, varicocele appears in the left scrotum, the reason being the anatomical dissimilarity of the two sides. In patients with right-sided or bilateral varicocele, further investigation to rule out a retroperitoneal pathology, especially neoplastic, is indicated. On rare occasions, however, a vascular anomaly as the underlying pathology must be considered. A patient is presented with a complete situs inversus accompanied by a right-sided varicocele. Intraoperative right internal spermatic phlebography revealed that this vein drained into the right renal vein, and valvular incompetence at their confluence was the etiology of the disease. Intraoperative internal spermatic phlebography in patients with right-sided or bilateral varicoceles is recommended as the simplest and safest means to demonstrate the etiology of this disease.


Controlled trial of bright light and negative air ions for chronic depression.
This randomized controlled trial investigates the efficacy of two non-pharmacologic treatments, bright light and high-density negative air ions for non-seasonal chronic depression. Both methods have shown clinical success for seasonal affective disorder (SAD). Patients were 24 (75%) women and 8 (25%) men, ages 22-65 years (mean age +/- S.D., 43.7 +/- 12.4 years), with Major Depressive Disorder, Single Episode (DSM-IV code, 296.2), Chronic (episode duration > or = 2 years). Patients were entered throughout the year and randomly assigned to exposure to bright light (10 000 lux, n = 10), or high-density (4.5 x 10(14) ions/s flow rate, n = 12) or low-density (1.7 x 10(11) ions/s, n = 10, placebo control) negative air ions. Home treatment sessions occurred for 1 h upon awakening for 5 weeks. Blinded raters assessed symptom severity weekly with the Structured Interview Guide for the Hamilton Depression Rating Scale--Seasonal Affective Disorder (SIGH-SAD) version. Evening saliva samples were obtained before and after treatment for ascertainment of circadian melatonin rhythm phase. SIGH-SAD score improvement was 53.7% for bright light and 51.1% for high-density ions v. 17.0% for low-density ions. Remission rates were 50%, 50% and 0% respectively. The presence or severity of atypical symptoms did not predict response to either treatment modality, nor were phase advances to light associated with positive response. Both bright light and negative air ions are effective for treatment of chronic depression. Remission rates are similar to those for SAD, but without a seasonal dependency or apparent mediation by circadian rhythm phase shifts. Combination treatment with antidepressant drugs may further enhance clinical response.


Advantages of using the species Solanum chacoense in breeding virus X-resistant potato forms.
The results of checking the resistance of diploid and polyploid specimens of the species Solanum chacoense to virus X are presented. The advantage of using this species in breeding resistant forms is shown.


Differential effects of stress-related and stress-unrelated humor in remitted depression.
Enhancing emotion regulation among previously depressed people is crucial for improving their resilience and reducing relapse. Therefore, emphasis is placed on determining effective regulation strategies, particularly those that, besides down-regulating negative emotions, also up-regulate positive emotions. One promising strategy, with great potential in both these respects, is humor. It is unclear, however, what type of humor is most adaptive in remitted depression. This study compared two distinct humor-based strategies: stress-related humor and stress-unrelated humor. Outpatients with remitted depression (N = 94) participated in a randomized experiment evoking personal stress and the subsequent application of stress-related humor, stress-unrelated humor, or a non-humorous regulation. They repeatedly reported positive and negative emotions (at four time points) and experienced distress (at three time points). There were also assessments of selective attention, subsequent performance, effort, and intrusive thoughts. Unlike non-humorous regulation, humor-based strategies had adaptive consequences, both immediately and after a delay; however, stress-unrelated humor was most beneficial and was the only effective strategy when attention deficits were present. Humor, especially if unrelated to stressors, might broaden the repertoire of powerful emotion regulation strategies in remitted depression. Humorous focusing on distress can be detrimental for patients with attention impairment.Clinical trial registration: The study was registered under the number ISRCTN86314628 (20/09/2021).


A multi-Chinese blood center study testing serologic-negative donor samples for hepatitis C virus and human immunodeficiency virus with nucleic acid testing.
A multi-blood center study was conducted to evaluate a human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) multiplex nucleic acid testing (NAT) donor screening test and to determine the residual risk for HIV-1 and HCV infection. A commercially available HIV-1 and HCV assay (Procleix, Chiron Corp.) was used for simultaneous detection of HIV-1 RNA and HCV RNA on 89,647 unlinked donor samples. NAT was performed with pools of 16 samples that had passed all routine screening tests. Single-donor NAT was performed for samples that had been disqualified by any reactive screening test result(s). Anti-HCV (Ortho third-generation HCV enzyme immunoassay [EIA]), alanine aminotransferase, and HCV NAT (Roche COBAS Amplicor HCV test) confirmatory tests were used for HCV EIA-nonreactive, HCV NAT-reactive samples. Three HCV NAT yield cases and no HIV-1 yield cases were detected. The yield rate for HCV NAT was 3.4 per 10(5) (95 percent confidence interval [CI], 0.7-9.8). The estimated incidence rate for HCV is 24.2 per 100,000 person-years (95% CI, 3.4-88.0). If minipool NAT is added to routine donor screening, the residual risk for HCV is estimated to be reduced to 1 in 20.4x10(4) (95% CI, 1 in 5.2x10(4)-1 in 165.5x10(4)). The residual risk for transfusion-transmitted HCV infection is still relatively high in China. Incorporating NAT technology into blood donor screening would be estimated to reduce the residual risk of HCV infections eightfold over current EIA screening.


The contributions of handedness and working memory to episodic memory.
Past studies have independently shown associations of working memory and degree of handedness with episodic memory retrieval. The current study takes a step ahead by examining whether handedness and working memory independently predict episodic memory. In agreement with past studies, there was an inconsistent-handed advantage for episodic memory; however, this advantage was absent for working memory tasks. Furthermore, regression analyses showed handedness, and complex working memory predicted episodic memory performance at different times. Results are discussed in light of theories of episodic memory and hemispheric interaction.


Deficient priming activity of newborn cord blood-derived polymorphonuclear neutrophilic granulocytes with lipopolysaccharide and tumor necrosis factor-alpha triggered with formyl-methionyl-leucyl-phenylalanine.
Newborn infants are more susceptible to bacterial infections than adults. This susceptibility has been attributed to defects in humoral and cellular activity. Host cellular activity can be modified by factors produced by bacteria or the host in response to infection. We assessed the effect of two factors associated with gram-negative bacterial infection, lipopolysaccharide (LPS) and TNF-alpha, on polymorphonuclear neutrophilic granulocytes (PMN) obtained from adult or newborns (umbilical cord blood). PMN were primed in vitro with LPS (10 micrograms/L) or TNF-alpha (10(-9) M) for 45 min and then assessed, using a chemiluminescence (CL) assay as an indicator of oxidative radical production with formyl-methionyl-leucyl-phenylalanine as the trigger for CL initiation. CL activity of unprimed PMN was similar for adults and newborns (13.3 and 13.7 CL units, respectively). After priming with LPS, CL activity was increased to 43.4 CL units for PMN from adults but to only 17.6 CL units for PMN from newborns (p < 0.001, adults versus newborn increment). Priming of PMN with LPS was most effective when autologous plasma was present. Using FITC-conjugated LPS and a flow cytometry assay, we could demonstrate no difference between the binding affinity of LPS for adult and newborn PMN. However, formyl-methionyl-leucyl-phenylalanine binding studies indicated that adult PMN had a higher number of binding sites. TNF-alpha priming of newborn PMN was also ineffective. Adult PMN increased CL activity by 3.9-fold when primed with TNF-alpha, whereas newborn PMN increased by only 1.75-fold (p < 0.005). This priming deficiency was not attributable to TNF-alpha receptors because phycoerythrin-conjugated TNF-alpha was associated with PMN from adults and newborns equally. Thus, PMN from newborns are not primed effectively in vitro with LPS or TNF-alpha. This defect may contribute to neonatal susceptibility to bacterial infection.


Evidence for the interchangeability of an avoidance behavior and a negative occasion setter.
Recent research on avoidance behavior provided evidence that such behavior can function as a negative occasion setter. We tested this hypothesis further by investigating whether the modulatory function of a stimulus occasion setter transfers selectively to a relation previously modulated by an avoidance behavior, and whether the modulatory function of an avoidance behavior transfers selectively to a relation previously modulated by a stimulus occasion setter. The three experiments reported in this article provided evidence to support this hypothesis. Furthermore, the results of Experiment 1 suggested that the presence of counterconditioning trials is not a necessary condition for an avoidance behavior to function as a negative occasion setter. All three reported experiments support the occasion-setting account of avoidance behavior.


Total hip arthroplasty for failed internal fixation of femoral neck fracture: a retrospective study with 2-14 years' follow-up of 345 patients.
The purpose of this study was to analyze mid- to long-term outcomes of total hip arthroplasty (THA) following failed internal fixation of femoral neck fracture. This study retrospectively analyzed 345 patients with femoral neck fracture who underwent THA after failure of internal fixation at our hospital between January, 2003 and December, 2019. Patients older than 55 years (n = 175) and patients no older than 55 years (n = 170) were compared in terms of complications and survival rates during follow-up, which lasted a mean of 6 years. The two age groups showed similarly low incidence of complications and similarly long periods of survival without revision surgery. Only three younger patients and two older patients underwent revision surgery during follow-up. The two groups showed similarly high survival rates at the end of follow-up (> 93%). Younger patients showed significantly bettter Harris hip score at last follow-up (90.2 vs. 88.1 points, p < 0.001) without clinically significant difference, but they required THA significantly earlier after internal fixation (4.4 vs. 6.8 years, p < 0.001). THA after failed internal fixation of femoral neck fracture is a well tolerated and effective procedure in older and younger patients.


Physiological variables affecting collagen lattice contraction by human dermal fibroblasts.
Normal human dermal fibroblasts cultured in collagen lattices can compact that matrix by the process known as lattice contraction. That process is a model of the pathological one of scar contracture or wound contraction and is affected by several factors. Lattice contraction is promoted by the addition of adequate amounts of fetal bovine serum to the medium (maximum contraction with 10% serum). The process requires energy, of which glucose and pyruvate have been shown to be adequate sources. When glucose is used as the substrate, the major pathway of energy generation appears to be anaerobic metabolism. When pyruvate is the only substrate, aerobic metabolism may be crucial. The synthesis of DNA is not required for lattice contraction, while protein synthesis is, although the identities of the specific proteins are unknown. Impairment of calcium ion transport inhibits lattice contraction, and the specific inhibition of calmodulin-calcium interactions by W-7 blocks contraction. W-7 at a concentration of 6 x 10(-6) M blocks lattice contraction completely, while it has no effect at any lower concentration. Impairing dynamic microtubule activity impairs contraction. Disrupting microfilaments by cytochalasin B completely blocks lattice contraction. Microfilament function and calcium-calmodulin may be linked by a mechanism involving myosin-ATPase. The process of cell-mediated lattice contraction requires the production of energy, protein synthesis, and a functional cytoskeleton.


Chronic intraparaventricular nuclear administration of orexin A in male rats does not alter thyroid axis or uncoupling protein-1 in brown adipose tissue.
Orexin A, synthesised in the posterolateral hypothalamus, has widespread distribution including the paraventricular nucleus (PVN), which is rich in thyrotropin-releasing hormone (TRH) neurones. Nerve fibres in the PVN synapse on neurones that send polysynaptic projections to brown adipose tissue (BAT), which is important in thermogenesis. A number of observations suggests orexin A may be involved in regulation of metabolism and thermogenesis. We investigated the effect of orexin A injected intracerebroventricularly (ICV) on thyroid-stimulating hormone (TSH) and thyroid hormones in male rats. We then examined the effect of chronic iPVN injections of orexin A on plasma TSH and uncoupling protein-1 (UCP-1) protein in BAT. Orexin A (3 nmol) administered ICV significantly suppressed plasma TSH at 10 and 90 min. Orexin A (0.3 nmol) administered into the PVN twice daily for 3 days significantly increased day-time 2-h food intake, but did not significantly alter nocturnal food intake. Though chronic iPVN orexin A altered diurnal food intake, there was no effect on 24-h food intake or body weight. Furthermore, orexin A administered chronically into the PVN did not alter UCP-1 level in BAT, or plasma hormones relative to saline injected animals. Chronic iPVN orexin A does not appear to influence thermogenesis through activation of UCP-1 or the thyroid axis.


One-Pot Purification and Immobilization of Phenylalanine Dehydrogenase from Bacillus nanhaiensi by Functional Reduced Graphene Oxide.
The one-pot immobilization of halophilic phenylalanine dehydrogenase from marine microorganism with metal ions modified reduced graphene oxide (CRGO) material was studied. Phenylalanine dehydrogenase was from Bacillus nanhaiensi and expressed with a C-terminal His-tag. Investigation of CRGO, CRGO-PEI, CRCO-Mn, and CRGO-PEI-Mn for one-pot purification and immobilization of phenylalanine dehydrogenase from crude enzyme solution was carried out. Enzyme activity yield rate achieved 80.0% by immobilization with CRCO-Mn, and the loading capacity was 6.7 mg/mg. Manganese ion coordination greatly improved the selectivity of the CRGO for the target His-tagged enzyme. Furthermore, the effect of NaCl concentration on the immobilization was investigated, which the loading capacity of CRGO-PEI and CRGO-Mn-PEI was increased by 10.7% and 30.6% with 1 M NaCl, respectively. The adsorption curves of crude enzyme one-pot immobilized by CRGO-Mn and purified enzyme immobilized by CRGO-Mn were similar. Therefore, one-pot immobilization strategy is promising for industrial application with advantages such as high efficiency and low cost, which shorten the pipelines for enzyme discovery towards industrial applications through the establishing of marine enzyme collections.


ReporTree: a surveillance-oriented tool to strengthen the linkage between pathogen genetic clusters and epidemiological data.
Genomics-informed pathogen surveillance strengthens public health decision-making, playing an important role in infectious diseases' prevention and control. A pivotal outcome of genomics surveillance is the identification of pathogen genetic clusters and their characterization in terms of geotemporal spread or linkage to clinical and demographic data. This task often consists of the visual exploration of (large) phylogenetic trees and associated metadata, being time-consuming and difficult to reproduce. We developed ReporTree, a flexible bioinformatics pipeline that allows diving into the complexity of pathogen diversity to rapidly identify genetic clusters at any (or all) distance threshold(s) or cluster stability regions and to generate surveillance-oriented reports based on the available metadata, such as timespan, geography, or vaccination/clinical status. ReporTree is able to maintain cluster nomenclature in subsequent analyses and to generate a nomenclature code combining cluster information at different hierarchical levels, thus facilitating the active surveillance of clusters of interest. By handling several input formats and clustering methods, ReporTree is applicable to multiple pathogens, constituting a flexible resource that can be smoothly deployed in routine surveillance bioinformatics workflows with negligible computational and time costs. This is demonstrated through a comprehensive benchmarking of (i) the cg/wgMLST workflow with large datasets of four foodborne bacterial pathogens and (ii) the alignment-based SNP workflow with a large dataset of Mycobacterium tuberculosis. To further validate this tool, we reproduced a previous large-scale study on Neisseria gonorrhoeae, demonstrating how ReporTree is able to rapidly identify the main species genogroups and characterize them with key surveillance metadata, such as antibiotic resistance data. By providing examples for SARS-CoV-2 and the foodborne bacterial pathogen Listeria monocytogenes, we show how this tool is currently a useful asset in genomics-informed routine surveillance and outbreak detection of a wide variety of species. In summary, ReporTree is a pan-pathogen tool for automated and reproducible identification and characterization of genetic clusters that contributes to a sustainable and efficient public health genomics-informed pathogen surveillance. ReporTree is implemented in python 3.8 and is freely available at https://github.com/insapathogenomics/ReporTree .


Music of infant-directed singing entrains infants' social visual behavior.
Infant-directed singing is a culturally universal musical phenomenon known to promote the bonding of infants and caregivers. Entrainment is a widely observed physical phenomenon by which diverse physical systems adjust rhythmic activity through interaction. Here we show that the simple act of infant-directed singing entrains infant social visual behavior on subsecond timescales, increasing infants' looking to the eyes of a singing caregiver: as early as 2 months of age, and doubling in strength by 6 months, infants synchronize their eye-looking to the rhythm of infant-directed singing. Rhythmic entrainment also structures caregivers' own cueing, enhancing their visual display of social-communicative content: caregivers increase wide-eyed positive affect, reduce neutral facial affect, reduce eye motion, and reduce blinking, all in time with the rhythm of their singing and aligned in time with moments when infants increase their eye-looking. In addition, if the rhythm of infant-directed singing is experimentally disrupted-reducing its predictability-then infants' time-locked eye-looking is also disrupted. These results reveal generic processes of entrainment as a fundamental coupling mechanism by which the rhythm of infant-directed singing attunes infants to precisely timed social-communicative content and supports social learning and development.


New York 1 and Sin Nombre viruses are serotypically distinct viruses associated with hantavirus pulmonary syndrome.
New York 1 virus (NY-1) and Sin Nombre virus (SN) are associated with hantavirus pulmonary syndrome (HPS). NY-1 and SN are derived from unique mammalian hosts and geographic locations but have similar G1 and G2 surface proteins (93 and 97% identical, respectively). Focus reduction neutralization assays were used to define the serotypic relationship between NY-1 and SN. Sera from NY-1-positive Peromyscus leucopus neutralized NY-1 and SN at titers of >/=1/3,200 and </=1/400, respectively (n = 12). Conversely, SN-specific rodent sera neutralized NY-1 and SN at titers of <1/400 and 1/6,400, respectively (n = 13). Acute-phase serum from a New York HPS patient neutralized NY-1 (1/640) but not SN (<1/20), while sera from HPS patients from the southwestern United States had 4- to >16-fold-lower neutralizing titers to NY-1 than to SN. Reference sera to Hantaan, Seoul, and Prospect Hill viruses also failed to neutralize NY-1. These results indicate that SN and NY-1 define unique hantavirus serotypes and implicate the presence of additional HPS-associated hantavirus serotypes in the Americas.


Tumors of the ear canal.
This discussion will focus on the clinical presentation, diagnostic evaluation, expected biologic behavior, and therapeutic options for tumours of the ear canal in the dog and cat. Particular emphasis will be placed on neoplasia of the ceruminous glands, squamous cell carcinoma, and non-neoplastic mass lesions.


Hazard evaluation of diisononyl phthalate and diisodecyl phthalate in a Japanese medaka multigenerational assay.
Reproductive and developmental effects of diisononyl phthalate (DINP) and diisodecyl phthalate (DIDP) were evaluated in a Japanese medaka (Oryzias latipes) multigeneration protocol. Each phthalate was administered via fish flake diets at a concentration of 20 microg/g (1 microg/g fish/day). Two controls were included, untreated and acetone carrier. The F(0) and F(1) generation adults were reared to sexual maturation and the test was ended prior to sexual maturation of the F(2) generation. Biochemical, individual, and population parameters were evaluated: testosterone metabolism, 7-ethoxyresorufin-o-deethylase (EROD) activity, survival, development, growth, gonadal-somatic index, histopathology, sex ratio, and fecundity. Male fish showed a two-fold induction of several testosterone metabolites in the DINP-treated group compared to the untreated control but not the acetone control. In a similar manner, in female fish only the DIDP-treated group expressed greater testosterone hydroxylase activity. There were neither sex- nor treatment-related differences in the results from the EROD assay. A statistically significant transient delay in red blood cell pigmentation was observed. The male-to-female ratio was consistent across treatments and the phenotypic and histological gender classifications were in agreement. Egg production was not significantly different among treatment groups. Neither phthalate elicited an effect on reproduction or development at various levels of biological organization.


Electrode kinetics at carbon electrodes and the density of electronic states.
Marcus-Hush theory relates the rate of electron transfer to the density of electronic states of the electrode material. Through use of a carbon microelectrode--for which the density of states is expected to vary as a function of potential--this predication is validated for graphitic materials by measurement of a variety of outer-sphere redox systems.


Carbetocin versus oxytocin to reduce additional uterotonic use at non-elective caesarean section: a double-blind, randomised trial (.).
We compared the efficacy of Carbetocin (long-acting oxytocin receptor agonist) versus Oxytocin given at non-elective caesarean section. We performed a double-blind, randomised, single-centre study. Eligible women were ≥37 weeks of gestation undergoing emergency caesarean section. Participants received either carbetocin of 100 μg or oxytocin 5 international units. The primary outcome was the need to administer additional uterotonics, as determined by the clinician. Secondary outcomes included estimated blood loss, haemoglobin drop pre-post operation and the need for a blood transfusion Results: From August 2012 to February 2013, 114 women were enroled. Two were excluded from analysis as they received a general anaesthetic. Fifty-nine patients received 100-μg carbetocin; 53 received 5 international units oxytocin. There was no statistically significant difference in the number of women requiring additional uterotonics between the two groups: Carbetocin group 22% and Oxytocin group 13% (p = 0.323). There were no significant differences in the fall in haemoglobin, estimated blood loss, rates of post-partum haemorrhage or blood transfusions. Oxytocin and carbetocin have similar requirements for additional uterotonics, estimated blood loss, haemoglobin drop and blood transfusions. There was a trend towards requiring additional uterotonics in patients receiving Carbetocin which was not statistically significant. This study found no benefits in using carbetocin over oxytocin.


Endothelial cell damage after intracapsular cataract extraction and primary anterior chamber pseudophakos implantation.
There is a gradual reduction in endothelial cell count with increasing age, but there is no significant difference in endothelial cell loss between aphakic eyes and pseudophakic eyes, the percentage being 33.14 and 26.28%, respectively.


Preoperative Breast Magnetic Resonance Imaging in Women With Local Ductal Carcinoma in Situ to Optimize Surgical Outcomes: Results From the Randomized Phase III Trial IRCIS.
We evaluated the addition of breast magnetic resonance imaging (MRI) to standard radiologic evaluation on the re-intervention rate in women with ductal carcinoma in situ (DCIS) undergoing breast-conserving surgery. Women with biopsy-proven DCIS corresponding to a unifocal microcalcification cluster or a mass less than 30 mm were randomly assigned to undergo MRI or standard evaluation. The primary end point was the re-intervention rate for positive or close margins (< 2 mm) in the 6 months after randomization ( ClinicalTrials.gov identifier: NCT01112254). A total of 360 patients from 10 hospitals in France were included in the study. Of the 352 analyzable patients, 178 were randomly assigned to the MRI arm, and 174 were assigned to the control arm. In the intent-to-treat analysis, 82 of 345 patients with the assessable end point were reoperated for positive or close margins within 6 months, resulting in a re-intervention rate of 20% (35 of 173) in the MRI arm and 27% (47 of 172) in the control arm. The absolute difference of 7% (95% CI, -2% to 16%) corresponded to a relative reduction of 26% (stratified odds ratio, 0.68; 95% CI, 0.41 to 1.1; P = .13). When considering only the per-protocol population with an assessable end point, the difference was 9% (stratified odds ratio, 0.59; 95% CI, 0.35 to 1.0; P = .05). Total mastectomy rates were 18% (31 of 176) in the MRI arm and 17% (30 of 173) in the control arm (stratified P = .93). For 100 lesions seen on MRI, nonmass-like enhancement was more predominant (82%) than mass enhancement (20%). Nevertheless, no specific morphologic and kinetic parameters for DCIS were identified. The study did not show sufficient surgical improvement with the use of preoperative MRI to be clinically relevant in DCIS staging. However, this could be reconsidered with the improvement of new MRI sequences and new modalities in magnetic resonance techniques.


A European nano-registry as a reliable database for quantitative risk assessment of nanomaterials? A comparison of national approaches.
Despite the fact that nanomaterials have been in use for decades and chemicals legislation is largely harmonised within the EU, quantitative and safety-relevant information on nanomaterials is still scarce. In particular, information about production volumes, their unique physicochemical properties (size, specific surface area, etc.) and nanomaterial exposure, which may lead to adverse effects on human health and the environment, is still lacking. While the latest amendments of the REACH Annexes have led to certain improvements, a harmonised EU-wide nano-registry would provide additional quantitative data for risk assessment but is not foreseeable for the near future. Since the European Commission, the European Parliament and some member states take contrasting approaches to the regulation of nanomaterials, France, Belgium, Denmark, Sweden and Norway (as a country of the European Economic Area), launched national mandatory reporting systems to collect quantitative information, thus fostering early risk assessment of nanomaterials. In this study, we compare national registries - based on a literature review and expert interviews - and show differences between the regulations under the respective national laws and REACH regulation. These include, for instance, thresholds for notification and level of detail on the specification of the nanomaterial, mixture and/or product, the definition of exceptions for the requirement to register and the timing of registration. As this heterogenous regulatory framework hinders comparability and potentially creates trade barriers, we argue that a harmonised EU-wide nano-registry would substantially improve the current situation by promoting the safe and sustainable handling of nanomaterials, increasing transparency and trust, and consequently nurturing innovation. Such an EU-wide nano-registry should both cover nanomaterials as substances or mixtures, such as in REACH registration, and the semi-/finished products they will be used in, since the exposure, and thus the hazardous potential of released nanomaterials during their life cycle, depends largely on the scope of application.


Cocrystals "Divorce and Marriage": When a Binary System Meets an Active Multifunctional Synthon in a Ball Mill.
A well-defined and stable "AB" binary system in the presence of "C" a crystalline synthon ground in a ball mill undergoes selective transformation in the solid state according to the equation AB+C→AC+B. When the amount of C is increased two times then the equation AB+2C→AC+BC is valid. The other variants are more complex. The pathway BC+A is allowed and leads to the AC and B products. The pathway AC+B is not preferred, and no transformation is observed. These non-obvious correlations were observed for cocrystal of barbituric acid (BA):thiobarbituric acid (TBA) recently reported by Shemchuk et al. (Chem. Commun. 2016, 52, 11815-11818) in the presence of 1-hydroxy-4,5-dimethyl-imidazole 3-oxide (HIMO). This synthon shows high affinity for the BA0.5 TBA0.5 cocrystal as well for its individual components, BA and TBA. Single-quantum, double-quantum (SQ-DQ) 2D 1 H very fast MAS NMR with a spinning rate of 60 kHz was employed as a basic and most diagnostic tool for the study of cocrystals transformations. Analysis of the experimental data was supported by theoretical calculations, including computation of the stabilization energy, Estab , defined as the energy difference between the energy of a co-crystal and the sum of the energies of particular components in the respective stoichiometric ratios. Two mechanisms of synthon replacement have been proposed. Pathway 1 assumes a concerted mechanism of substitution. In this approach, synthon attack is synchronized in time with the departure of one of the components of the binary system. Pathway 2 implies a non-concerted process, with an intermediate stage in which three separate components are present. Evidence suggesting a preference for Pathway 2 is shown.


Exosome-based strategies for diagnosis and therapy of glioma cancer.
Glioblastoma belongs to the most aggressive type of cancer with a low survival rate that is characterized by the ability in forming a highly immunosuppressive tumor microenvironment. Intercellular communication are created via exosomes in the tumor microenvironment through the transport of various biomolecules. They are primarily involved in tumor growth, differentiation, metastasis, and chemotherapy or radiation resistance. Recently several studies have highlighted the critical role of tumor-derived exosomes against immune cells. According to the structural and functional properties, exosomes could be essential instruments to gain a better molecular mechanism for tumor understanding. Additionally, they are qualified as diagnostic/prognostic markers and therapeutic tools for specific targeting of invasive tumor cells such as glioblastomas. Due to the strong dependency of exosome features on the original cells and their developmental status, it is essential to review their critical modulating molecules, clinical relevance to glioma, and associated signaling pathways. This review is a non-clinical study, as the possible role of exosomes and exosomal microRNAs in glioma cancer are reported. In addition, their content to overcome cancer resistance and their potential as diagnostic biomarkers are analyzed.


Molecular characterisation of the causative agents of Cryptococcosis in patients of a tertiary healthcare facility in the state of Amazonas-Brazil.
As there are four major molecular types of Cryptococcus neoformans (VNI, VNII, VNIII and VNIV) and four molecular types of Cryptococcus gattii (VGI, VGII, VGIII and VGIV), it is important to identify the specific groups causing cryptococcosis in different geographical regions. Here, we investigated the molecular types of 57 cryptococcal isolates from patients in a tertiary care hospital in the state of Amazonas, Brazil, between 2006 and 2010. The isolates were characterised by PCR fingerprinting using the M13 minisatellite and confirmed by URA5-RFLP analysis, and the presence of specific genes from the mating type locus (MATα and MATa) of these species was analysed by PCR. Most of the patients were male (66.7%), between 16 and 30 years of age (51.7%), and HIV-positive (75.0%). Most isolates were collected from cerebrospinal fluid samples (71.7%). Most of the C. neoformans isolates (n=40) were characterised as members of the VNI molecular group (n=39), a unique isolate was characterised as VNII whereas all isolates of C. gattii (n=17) were members of the VGII molecular group. With regard to mating types, 55 isolates were type 'α', and only two were type 'a'. This study revealed the prevalence of the VNI molecular group and provides the first reported observation of the VNII molecular group in the northern region of Brazil.


Two-stage hepatectomy: who will not jump over the second hurdle?
Two-stage hepatectomy uses compensatory liver regeneration after a first noncurative hepatectomy to enable a second curative resection in patients with bilobar colorectal liver metastasis (CLM). To determine the predictive factors of failure of two-stage hepatectomy. Between 2000 and 2010, 48 patients with irresectable CLM were eligible for two-stage hepatectomy. The planned strategy was a) cleaning of the left hepatic lobe (first hepatectomy), b) right portal vein embolisation and c) right hepatectomy (second hepatectomy). Six patients had occult CLM (n = 5) or extra-hepatic disease (n = 1), which was discovered during the first hepatectomy. Thus, 42 patients completed the first hepatectomy and underwent portal vein embolisation in order to receive the second hepatectomy. Eight patients did not undergo a second hepatectomy due to disease progression. Upon univariate analysis, two factors were identified that precluded patients from having the second hepatectomy: the combined resection of a primary tumour during the first hepatectomy (p = 0.01) and administration of chemotherapy between the two hepatectomies (p = 0.03). An independent association with impairment to perform the two-stage strategy was demonstrated by multivariate analysis for only the combined resection of the primary colorectal cancer during the first hepatectomy (p = 0.04). Due to the small number of patients and the absence of equivalent conclusions in other studies, we cannot recommend performance of an isolated colorectal resection prior to chemotherapy. However, resection of an asymptomatic primary tumour before chemotherapy should not be considered as an outdated procedure.


Pericardial fat necrosis.
Pericardial fat necrosis has a highly characteristic clinical picture that enables a preoperative diagnosis to be made on clinical grounds. Surgical therapy, which remains the treatment of choice for this curable entity, confirms the diagnosis. This report reviews the literature and adds a thirteenth documented case to the previously reported 12 cases.


Well-differentiated adenoma at the anorectal junction.
Twenty-six cases of patients having adenomatous polyps at the anorectal junction are presented. Most polyps in this region were well-differentiated, and atypical polyps were infrequently seen. Well-differentiated adenomatous polyps in the anorectal junction were larger in average size than ordinary colorectal polyps having the same grade of epithelial pseudostratification. The reason adenomatous polyps at the anorectal junction grow to large sizes with only a relatively rare occurrence of malignant degeneration is discussed in relation to their specific location. Some well-differentiated adenomas were considered to develop secondarily in the bases of hemorrhoids or in pre-existing fibrous polyp that were the end result of hemorrhoids.


Correction: Kinetics of antibody responses to PfRH5-complex antigens in Ghanaian children with Plasmodium falciparum malaria.
[This corrects the article DOI: 10.1371/journal.pone.0198371.].


[Study on the function of HSD-3.8 gene encoding a testis-specific protein with yeast two-hybrid system].
To explore the protein factors that could interact with the testis-specific protein encoded by HSD-3.8 gene (GenBank Accession Number AF311312) related with female fertilization. Yeast two-hybrid system was used to screen the human ovary MATCHMAKER cDNA library with constructed "bait plasmid" containing the 0.7 kb fragment (HSD-0.7) of HSD-3.8. The interaction with the positive fragments using a series of truncated bait plasmids was investigated. One positive gene fragment was obtained, which coded for 144 amino acids of the C-terminus of human G protein beta subunit 1. Truncated bait plasmids couldn't interact with the fish protein fragment in yeast. The protein encoded by HSD-3.8 gene may function through G protein signal transduction pathway and the interaction depends on the integration of the bait protein.


Behavioral effects of xylamine-induced depletions of brain norepinephrine: interaction with LSD.
Male rats were treated with a combination of systemic fluoxetine and intraventricular xylamine (under ether anesthesia) to deplete brain norepinephrine (NE) in the projection areas of the locus coeruleus. Four days later, control and lesioned rats were tested following injections of either saline or 80 micrograms/kg LSD in a Behavioral Pattern Monitor which recorded the sequential patterns of their locomotor and investigatory (holepokes) responses. Liquid chromatographic measures of brain monoamines confirmed that xylamine reduced hippocampal NE by 80.8% and hypothalamic NE by 26% without affecting levels of serotonin or dopamine. Relative to controls, NE-depleted rats exhibited repetitive spatial patterns of locomotion with no alteration in the amount or rate of habituation of locomotor activity. Lesioned animals made fewer rearings and holepokes, particularly early in the hour test session. When given 80 micrograms/kg LSD, sham-lesioned rats exhibited the expected decreases in entries into and time spent in the center of the chamber, an increase in time spent in the corners, and fewer holepokes and rearings early in the session. With the exception of the effect on rearings and holepokes, the effects of LSD were diminished in rats depleted of brain NE. These results indicate that this profile of behavioral effects of LSD, which has been interpreted as a potentiation of neophobia, may be dependent upon the noradrenergic projections of the locus coeruleus.


Analysis of the autoimmune epitopes on human testicular NASP using recombinant and synthetic peptides.
The human nuclear autoantigenic sperm protein, NASP, is a testicular histone-binding protein of 787 amino acids to which most vasectomized men develop autoantibodies. In this study to define the boundaries of antigenic regions and epitope recognition pattern, recombinant deletion mutants spanning the entire protein coding sequence and a human NASP cDNA sublibrary were screened with vasectomy patients' sera. Employing panel sera from 21 vasectomy patients with anti-sperm antibodies, a heterogeneous pattern of autoantibody binding to the recombinant polypeptides was detected in ELISA and immunoblotting. The majority of sera (20/21) had antibodies to one or more of the NASP fusion proteins. Antigenic sites preferentially recognized by the individual patients' sera were located within aa 32-352 and aa 572-787. Using a patient's serum selected for its reactivity to the whole recombinant protein in Western blots, cDNA clones positive for the C-terminal domain of the molecule were identified. The number and location of linear epitopes in this region were determined by synthetic peptide mapping and inhibition studies. The epitope-containing segment was delimited to the sequence aa 619-692 and analysis of a series of 74 concurrent overlapping 9mer synthetic peptides encompassing this region revealed four linear epitopes: amino acid residues IREKIEDAK (aa 648-656), KESQRSGNV (aa 656-664), AELALKATL (aa 665-673) and GFTPGGGGS (aa 680-688). All individual patients' sera reacted with epitopes within the sequence IREellipsis.GGS (aa 648-688). The strongest reactivity was displayed by peptides corresponding to the sequence AELALKATL (aa 665-673). Thus, multiple continuous autoimmune epitopes in NASP involving sequences in the conserved C-terminal domain as well as in the less conserved testis-specific N-terminal region comprising the histone-binding sites, as predicted for an antigen-driven immune response, may be a target of autoantibodies in vasectomized men and may provide a relevant laboratory variable to describe more accurately the spectrum of autoantibody specificities associated with the clinical manifestation of vasectomy.


Effect of increased circulating corticosterone in the immature fowl on the blastogenic responses of peripheral blood lymphocytes.
Plasma corticosterone concentrations were increased by giving chickens implants containing corticosterone and the effects on lymphocyte numbers and lymphocyte responses to the mitogens, phytohaemagglutinin (PHA) and concanavalin A (con A), were measured. Raising plasma corticosterone caused a dose-related decrease in numbers of circulating lymphocytes. Blastogenic responses of lymphocytes in whole blood to PHA and con A were reduced and stimulation indices were decreased in those birds with raised circulating corticosterone. When the initial corticosterone-induced lymphocytopenia was taken into account it was evident that corticosterone depressed the responsiveness of the lymphocytes to PHA but not to con A. Additions of corticosterone to cultures of whole blood showed that PHA stimulation was more sensitive than con A stimulation to the inhibitory effects of corticosterone.


Multiple sclerosis and the workplace: report of an industry-based cluster.
Eleven cases of MS occurred within a 10-year period in a zinc-related manufacturing plant. The observed disease incidence was greater than expected from population data, using multiple approaches to statistical analysis (p less than or equal to 0.01). A case-control study, performed to examine several zinc parameters in blood, failed to indicate specific abnormalities among the MS patients, but all subjects (both MS and controls) working in the plant demonstrated higher serum zinc levels than all subjects (MS and controls) not working there.


Predictive factors of outcome after liver transplantation in patients with cirrhosis and hepatocellular carcinoma.
Studies to define the optimal upper limits of tumor size and number as predictors of outcome after orthotopic liver transplantation (OLT) have yielded conflicting results. We analyzed 72 patients with cirrhosis and hepatocellular carcinoma (HCC) who underwent OLT over a 12-year period in a single center. Predictive factors for survival and tumor recurrence, according to the Milan criteria, were also examined. Our cohort included 60 men and 12 women of mean age 54 +/- 8 years and mean follow-up of 40 +/- 39 months. Origin of cirrhosis was postviral in 70% and Child class B or C in two thirds of patients. HCC was multifocal in 61%; about one fifth of patients had micro- or macrovascular involvement or positive nodes upon histologic examination. The cumulative size of the lesions was <3 cm in 17 patients; >3 to < or =5 cm in 28 patients; >5 to < or =8 cm in 14 patients; and >8 cm in 13 patients. According to the number and size of tumor nodules, 49 patients met the Milan criteria. During follow-up 25 patients died, 13 due to tumor recurrence. The 1- and 2-year survivals were 90% and 85% for patients who met the Milan criteria versus 57% and 51% for patients exceeding those limits (P = .006). A cumulative tumor size >8 cm was predictive of survival and tumor recurrence upon multivariate analysis. The adoption of Milan criteria for selection of cirrhotic patients has improved survival and reduced the rate of tumor recurrence. The evaluation of cumulative tumor size might further improve patient selection.


Acute renal infarction from a cardiac thrombus.
A 53-year-old man presented to hospital 2 hours after the abrupt onset of left upper abdominal pain. He was treated with analgesics and discharged after 4 hours of observation, but presented to another hospital 2 hours later with severe left abdominal pain. His past medical history included ischemic dilated cardiomyopathy due to recurrent myocardial infarction. Physical examination, electrocardiography, laboratory investigations, contrast-enhanced computed tomography, and transesophageal echocardiography. Renal artery thromboembolism resulting from dilated cardiomyopathy, severely reduced cardiac function and an intracardiac thrombus. Anticoagulation with unfractionated heparin followed by enoxaparin and warfarin.


"Oxidative stress induced by phthalates in mammals: State of the art and potential biomarkers".
Phthalates, plasticizers that are widely used in consumer products including toys, cosmetics, and food containers, have negative effects in liver, kidney, brain, lung and reproductive system of humans and other mammals. To summarize, describe and discuss the available information on the effects of phthalate exposure in mammals, with emphasis on oxidative stress, and to suggest potential biomarkers of the health risks associated with phthalate exposure. An assessment of scientific journals was performed using the PRISMA model for systematic reviews. Manuscripts reporting effects of phthalate exposure on mammalian health published in the last decade were selected according to originality, content, and association to health hazards. We identified 25 peer-reviewed articles published between January 1st, 2010 and June 1st, 2021 that fit the aims and selection criteria. Phthalates induce oxidative stress and cell degenerative processes by increasing intracellular reactive species. Antioxidant cytoprotective systems decrease with time of exposure; conversely, oxidative damage markers, including thiobarbituric acid-reactive substances (TBARS), 8-hydroxy-2'-desoxyguanosine (8-OHdG) and malondialdehyde (MDA), increase. Phthalates were associated with endocrine system disfunction, metabolic disorders, infertility, nonviable pregnancy, cell degeneration, growth impairment, tumor development, and cognitive disorders. Phthalates can also aggravate health conditions such as asthma, hepatitis, diabetes, allergies, chronic liver and kidney diseases. Among humans, the more vulnerable subjects to phthalate exposure effects were children and individuals with a prior health condition. Chronic exposure to phthalates induces oxidative stress in mammals with concomitant adverse effects in reproductive, respiratory, endocrine, circulatory, and central nervous systems in both in vitro and in vivo trials. Oxidative damage markers and phthalate metabolites levels were the most common biomarkers of phthalate exposure effects. Studies in free-ranging and wild mammals are nil. Further studies on the pathways that lead to metabolic disruption are needed to identify potential treatments against phthalate-induced detrimental effects.


Turkey beta 2-microglobulin--II. Physiological parameters investigated by radioimmunoassay.
A competitive binding radioimmunoassay (RIA) was developed utilizing 125I-labeled turkey beta 2-microglobulin (beta 2m), rabbit anti-turkey beta 2m serum and polyethylene glycol for separation of bound and free antigen. The antibody-antigen reaction was demonstrated to be monospecific by immunofixation electrophoresis. Characteristic parameters of the RIA were: nonspecific binding, 5.5%; linear dose-response range, 0.24-64.0 ng turkey beta 2m; dose-response correlation coefficient, -0.995; assay sensitivity, 0.52 ng; upper limit of precision, 40.4 ng. RIA analysis of adult turkey tissue and organ homogenates revealed highest amounts (greater than 10 micrograms turkey beta 2m/g) in liver and kidney and intermediate levels (2-10 micrograms/g) in lymphoid organ (thymus, spleen, caecal tonsil, bursa), skin and lung extracts. Erythrocytes and peripheral blood lymphocyte extracts contained 8.0 and 5.1 micrograms/10(9) cells respectively whereas all other tissues examined expressed greater than 2.0 micrograms/g. The beta 2m content in turkey serum was 192.5 micrograms/ml. The molecular weight distribution of turkey beta 2m in serum determined by gel permeation chromatography consisted of a minor component of approximately 60,000 daltons and a major fraction eluting at a position characteristic of free beta 2m. A cross-reaction between turkey and mammalian beta 2ms was not observed.


[Guidelines for type 1 Gaucher's disease].
Gaucher's disease (GD) is an inborn error of metabolism of the lysosomal enzyme beta-glucosidase, which induces the deposition of undegraded glycolipid material in organs rich in mononuclear macrophage system including liver, spleen and bone marrow. Beta-glucosidase is suspected in patients with evidence of liver and/or spleen enlargement, anemia and/or thrombocytopenia not attributable to other causes, or bone crisis especially in children. In scarce cases GD is associated with neurological involvement. The diagnosis confirmation is performed by analysis of beta-glucosidase activity in peripheral blood leukocytes or fibroblasts. It is necessary to identify genetic mutations that cause GD. It is also advisable to identify biomarkers including chitotriosidase activity and plasma CCL-18/PARC concentration. The use of a protocol for assessing the intensity of the deposit is mandatory to establish the indication for treatment, especially in rare diseases. Nowadays there are several options for treatment of GD: enzyme replacement therapy and substrate reduction therapy. Regular assessments are needed to establish the response and the degree of achievement of the therapeutic goals recommended through expert consensus.


Dual catalytic role of the metal ion in nickel-assisted peptide bond hydrolysis.
In our previous research we demonstrated the sequence specific peptide bond hydrolysis of the R1-(Ser/Thr)-Xaa-His-Zaa-R2 in the presence of Ni(II) ions. The molecular mechanism of this reaction includes an N-O acyl shift of the R1 group from the Ser/Thr amine to the side chain hydroxyl group of this amino acid. The proposed role of the Ni(II) ion is to establish favorable geometry of the reacting groups. In this work we aimed to find out whether the crucial step of this reaction--the formation of the intermediate ester--is reversible. For this purpose we synthesized the test peptide Ac-QAASSHEQA-am, isolated and purified its intermediate ester under acidic conditions, and reacted it, alone, or in the presence of Ni(II) or Cu(II) ions at pH 8.2. We found that in the absence of either metal ion the ester was quickly and quantitatively (irreversibly) rearranged to the original peptide. Such reaction was prevented by either metal ion. Using Cu(II) ions as CD spectroscopic probe we showed that the metal binding structures of the ester and the final amine are practically identical. Molecular calculations of Ni(II) complexes indicated the presence of steric strain in the substrate, distorting the complex structure from planarity, and the absence of steric strain in the reaction products. These results demonstrated the dual catalytic role of the Ni(II) ion in this mechanism. Ni(II) facilitates the acyl shift by setting the peptide geometry, and prevents the reversal of the acyl shift, by stabilizing subsequent reaction products.


Human-algorithm teaming in face recognition: How algorithm outcomes cognitively bias human decision-making.
In face recognition applications, humans often team with algorithms, reviewing algorithm results to make an identity decision. However, few studies have explicitly measured how algorithms influence human face matching performance. One study that did examine this interaction found a concerning deterioration of human accuracy in the presence of algorithm errors. We conducted an experiment to examine how prior face identity decisions influence subsequent human judgements about face similarity. 376 volunteers were asked to rate the similarity of face pairs along a scale. Volunteers performing the task were told that they were reviewing identity decisions made by different sources, either a computer or human, or were told to make their own judgement without prior information. Replicating past results, we found that prior identity decisions, presented as labels, influenced volunteers' own identity judgements. We extend these results as follows. First, we show that the influence of identity decision labels was independent of indicated decision source (human or computer) despite volunteers' greater distrust of human identification ability. Second, applying a signal detection theory framework, we show that prior identity decision labels did not reduce volunteers' attention to the face pair. Discrimination performance was the same with and without the labels. Instead, prior identity decision labels altered volunteers' internal criterion used to judge a face pair as "matching" or "non-matching". This shifted volunteers' face pair similarity judgements by a full step along the response scale. Our work shows how human face matching is affected by prior identity decision labels and we discuss how this may limit the total accuracy of human-algorithm teams performing face matching tasks.


Uptake and use of recommendations for the diagnosis, severity scoring and management of chronic GVHD: an international survey of the EBMT-NCI Chronic GVHD Task Force.
In 2005, the National Institutes of Health (NIH) consensus conference published a series of papers recommending methods to improve the conduct of clinical trials in chronic GVHD. Although the NIH recommendations were primarily aimed at strengthening research, several papers addressed issues relevant for clinical practice, particularly diagnosis, severity scoring, and ancillary and supportive care practices. We conducted an international survey to assess the uptake of these recommendations, identify barriers to greater use and document the use and perceived effectiveness of available treatments. The response rate for the American survey of 1387 practitioners was 21.8%, and it was 24.6% for 407 centers surveyed in Europe, Asia, Australia and Africa. Most respondents were familiar with the NIH consensus recommendations (94-96%) and used them in practice. Multiple barriers to greater use were reported. Besides lack of time (55-62%), unfamiliarity with the recommendations, scarcity of evidence supporting the impact of recommendations on outcomes, insufficient training/experience in chronic GVHD management and inaccessibility of subspecialists were also endorsed. Systemic corticosteroids were reported to be the most effective treatment for chronic GVHD, but many others were perceived to have moderate or great success. Therapeutic management of steroid-refractory chronic GVHD was identified as the highest priority for research.


Is there standard chemotherapy for metastatic bladder cancer? Quality of life and medical resources utilization based on largest to date randomized trial.
A large multinational phase III trial performed during the late 1990s compared two systemic chemotherapy regimens gemcitabine plus cisplatin and methotrexate, vinblastine, doxorubicin, cisplatin (M-VAC) in more than 400 patients with advanced or metastatic urothelial cancer. This trial has been discussed to landmark the beginning of a new era following M-VAC polychemotherapy which has dominated treatment of advanced urothelial cancer throughout the previous decade. Despite the fact that gemcitabine/cisplatin combination therapy did not surpass M-VAC therapy in regard to patient survival as initially intended, this combination demonstrated a more favourable toxicity profile with improved tolerability and superior cost effectiveness, rendering this combination an attractive alternative to M-VAC. This review on the largest to date phase III trial for advanced or metastatic urothelial cancer will focus on issues of quality of life including indicators such as performance status and changes in body weight. Furthermore, data on medical resources utilization as accounted during systemic polychemotherapy and related toxic events will be reconsidered, particularly under the impression of decreasing health care resources worldwide.


Perivascular spaces mediate a relationship between diabetes and other cerebral small vessel disease markers in cerebrovascular and neurodegenerative diseases.
Type 2 diabetes mellitus (T2DM) and hypertension are risk factors for cerebral small vessel disease (SVD); however, few studies have characterised their relationships with MRI-visible perivascular spaces (PVS). MRI was used to quantify deep (d) and periventricular (p) white matter hyperintensities (WMH), lacunes, PVS in the white matter (wmPVS) or basal ganglia (bgPVS), and diffusion metrics in white matter. Patients with T2DM had greater wmPVS volume and there were greater wmPVS volumes in patients with T2DM and hypertension together. Counterfactual moderated mediation models found indirect effects of T2DM on volumes of other SVD and diffusion markers that were mediated by wmPVS: pWMH, dWMH, periventricular lacunes, and deep lacunes, and progression of deep lacunes over 1 year, in patients with hypertension, but not in patients without hypertension. Studying the regulation of cortical perivascular fluid dynamics may reveal mechanisms that mediate the impact of T2DM on cerebral small vessels.


Balancing the Anteroposterior Diameters of the Nostril Lengths in Cleft Rhinoplasty.
Osteocartilaginous deformities in cleft rhinoplasties may be restored with numerous techniques. However, the anteroposterior (AP) diameter lengths of the nostrils may still be unequal and should also be addressed. A technique was designed to balance nostril AP diameter lengths and apical shapes. Balance between AP nostril diameters was accomplished by reduction of the AP diameter of the nostril at the non-cleft side by medial crural reduction and augmentation of the AP diameter of the nostril at the cleft side by three-parted mini-flap reconstruction at the soft triangle. The cleft-side AP diameter length was divided by the non-cleft-side AP diameter length of the same preoperative, per-operative and 1-year postoperative base view photographs of each patient, and "nostril balancing ratio" was obtained. The more this ratio was near to "one," the more the AP nostril diameters were equal. Seventy-eight unilateral-cleft rhinoplasty were performed between January/2019 and May/2022. Forty-two of them required nostril AP diameter equalization. Twenty-nine patients were female, thirteen were male. Thirty-three of them were operated for a primary cleft rhinoplasty. Nine of them for secondary cleft rhinoplasty. Mean age was 28 years (22-39 years). Mean follow-up was 25 months (6-40 months). The preoperative, per-operative and postoperative mean "nostril balancing ratios" were 0.714 (0.621-0.813), 0.743 (0.721-0.752) and 0.971 (0.943-0.976), respectively. Balancing AP diameter symmetry with MCO at the non-cleft side and three-parted mini skin flap reconstruction at the cleft side may provide the satisfactory results. Three mini-flaps at the soft triangle may remold the nostril apex in an oval shape, which may result in a better shape symmetry. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Removal of microorganic pollutants in aquatic environment: The utilization of Fe(VI).
Microorganic pollutants (MOPs) in aquatic environment with low levels but high toxicity are harmful to ecosystem and human health. Fe(VI) has a dual-functional role in oxidation and coagulation, and can effectively remove MOPs, heavy metal, phosphate, particulates and colloids. Moreover, Fe(VI) can combine with traditional coagulants, or use as a pretreatment for membrane treatment because of its characters to generate nanoparticles by degradation in water. Based on the relevant toxicity experiments, Fe(VI) had been proved to be safe for the efficient treatment of MOPs. For better utilization of Fe(VI), its oxidation and coagulation mechanisms are summarized, and the knowledge about the control parameters, utilization methods, and toxicity effect for Fe(VI) application are reviewed in this paper. pH, different valences of iron, environmental substances, and other parameters are summarized in this study to clarify the important factors in the treatment of MOPs with Fe(VI). In the future study, aiming at cost reduction in Fe(VI) preparation, transportation and storage, enhancement of oxidation in the intermediate state, and better understanding the mechanism between interface and Fe(VI) oxidation will help promote the application of Fe(VI) in the removal of MOPs. This study offers guidelines for the application and development of Fe(VI) for the treatment of MOPs in aquatic environment.


Development of a bead-based multiplex assay to quantify bovine interleukin-10, tumor necrosis factor-α, and interferon-γ concentrations in plasma and cell culture supernatant.
The quantification of cytokines can improve our understanding of immune response and inflammation dynamics in dairy cows. Bead-based assays provide a sensitive, high-throughput platform, allowing for simultaneous quantification of multiple cytokines within a wide linear detection range. Our objective was to develop a multiplex bead-based assay using monoclonal antibodies for simultaneous quantification of bovine tumor necrosis factor (TNF)-α, IL-10, and IFN-γ in plasma and peripheral blood mononuclear cell (PBMC) culture supernatants. Recombinant cytokine standards produced in mammalian cells were used to determine the lower limit of detection and the linear detection range for each cytokine. The lower limit of detection was 110 pg/mL for IL-10, 95 pg/mL for TNF-α, and 20 pg/mL for IFN-γ. The linear quantification range was 110 to 241,000 pg/mL for IL-10, 95 to 620,000 pg/mL for TNF-α, and 20 to 130,000 pg/mL for IFN-γ. All 3 monoclonal capture and detection antibodies were specific for their respective cytokine analyte when using the recombinant IL-10, TNF-α, and IFN-γ standards. Intraassay and interassay coefficients of variation (CV) were <10% and <12%, respectively, for all analytes and samples matrices. Next, concentrations of native cytokines were determined in PBMC culture supernatants (n = 4) and in plasma from whole-blood samples (n = 6) with or without stimulation with Escherichia coli lipopolysaccharide or a mix of phorbol myristate acetate (PMA) and ionomycin. Peak concentrations of all 3 cytokines were secreted from PBMC after PMA/ionomycin stimulation (TNF-α, 8 h, range: 39,266-506,422 pg/mL; IL-10, 18 h, range: 15,770-63,415 pg/mL; IFN-γ 18 h, range: 189,977-492,659 pg/mL). In contrast, the highest concentrations in plasma from whole-blood stimulation were observed for IL-10 and TNF-α after LPS stimulation (TNF-α, 4 h, range: 1,764-13,460 pg/mL; IL-10, 24 h, range: 2,401-6,371 pg/mL), whereas PMA and ionomycin induced the highest secretion of IFN-γ (18 h, range: 53-20,215 pg/mL). In conclusion, the multiplex assay can quantify native IL-10, TNF-α, and IFN-γ across a broad concentration range in bovine plasma and cell culture supernatant, thereby providing a novel tool to evaluate inflammatory profiles in cattle and especially in dairy cows with inflammatory conditions. The existing multiplex assay can be expanded in the future by adding bead assays for additional bovine cytokines.


Comparative efficacy of sorafenib versus best supportive care in recurrent hepatocellular carcinoma after liver transplantation: a case-control study.
The efficacy of sorafenib in the post-liver transplantation (LT) setting has been scarcely studied. The aim of this study was to evaluate the efficacy of sorafenib, compared to best supportive care (BSC), in two cohorts of patients which presented with hepatocellular carcinoma (HCC) recurrence after LT. Data from patients who developed presentation or progression of HCC recurrence after LT not amenable to surgical/locoregional treatments (untreatable presentation/progression, UP) were retrieved. The cohort of patients receiving sorafenib starting from 2007 was compared to that of patients receiving BSC in the previous era. Disease outcome was investigated in terms of survival from recurrence or from UP by means of univariate and multivariate Cox regression models with event times left-truncated at the date of UP. Of a total of 39 patients, 24 received BSC and 15 sorafenib. The two groups were well matched at baseline, with time-related imbalances regarding mTOR-based immunosuppression and median time from LT to recurrence, significantly higher in the sorafenib group. Patients' outcome in the sorafenib group was significantly improved (median survival from recurrence 21.3 vs.11.8 months, HR=5.2, p=0.0009; median survival from UP 10.6 vs. 2.2 months, HR=21.1, p<0.0001). The only factor associated with survival after HCC recurrence in multivariate analysis was treatment with sorafenib (HR=4.0; p=0.0325). No severe adverse event was registered in this post-LT setting. Although the use of historical controls weakens final interpretation, sorafenib seems to be associated with an acceptable safety profile and benefit in survival in HCC patients suffering recurrence after LT.


Deep infiltrating endometriosis: Can magnetic resonance imaging anticipate the need for colorectal surgeon intervention?
To identify magnetic resonance imaging (MRI) features associated with colorectal surgical bowel resection for treatment of deep infiltrating endometriosis (DIE). 122 preoperative pelvic MRIs in women with laparoscopically-proven DIE and subsequent surgery (2006-2015) were identified, and retrospective cohort analysis performed. MRIs were reviewed independently by two radiologists blinded to surgical/histopathological outcomes. Associations between MRI characteristics of middle/posterior compartment endometriosis and surgical outcomes were investigated to identify MRI features associated with colorectal surgical bowel resection. MRI features associated with colorectal surgical intervention were: presence of an MRI bowel lesion (sensitivity 95.3%, specificity 63.3%, ROC-AUC 0.79); MRI bowel lesions ≥20 mm in length (sensitivity 91%, specificity 77%, ROC-AUC 0.84); MRI bowel lesions invading the muscularis or submucosa/mucosa layers (sensitivity 95.3%, specificity 63.3%, ROC-AUC 0.90). This study identifies MRI features that have potential diagnostic utility in identifying the need for colorectal surgical intervention in patients with DIE.


Autophagy and proteins involved in vesicular trafficking.
Autophagy is an intracellular degradation system that, as a basic mechanism it delivers cytoplasmic components to the lysosomes in order to maintain adequate energy levels and cellular homeostasis. This complex cellular process is activated by low cellular nutrient levels and other stress situations such as low ATP levels, the accumulation of damaged proteins or organelles, or pathogen invasion. Autophagy as a multistep process involves vesicular transport events leading to tethering and fusion of autophagic vesicles with several intracellular compartments. This review summarizes our current understanding of the autophagic pathway with emphasis in the trafficking machinery (i.e. Rabs GTPases and SNAP receptors (SNAREs)) involved in specific steps of the pathway.