Dynamic Changes in Circulating Tumor DNA During Chemoradiation for Locally Advanced Lung Cancer.
Concurrent chemoradiation therapy (CRT) is the principal treatment modality for locally advanced lung cancer. Cell death due to CRT leads to the release of cell-free DNA (cfDNA) and circulating tumor DNA (ctDNA) into the bloodstream, but the kinetics and characteristics of this process are poorly understood. We hypothesized that there could be clinically meaningful changes in cfDNA and ctDNA during a course of CRT for lung cancer. Multiple samples of plasma were obtained from 24 patients treated with CRT for locally advanced lung cancer to a mean dose of 66 Gy (range, 58-74 Gy) at the following intervals: before CRT, at weeks 2 and 5 during CRT, and 6 weeks after treatment. cfDNA was quantified, and a novel next generation sequencing (NGS) technique using enhanced tagged/targeted-amplicon sequencing was performed to analyze ctDNA. Patients for whom specific mutations in ctDNA were undetectable at the baseline time point had improved survival, and potentially etiologic driver mutations could be tracked throughout the course of CRT via NGS in multiple patients. We quantified the levels of cfDNA from patients before CRT, at week 2, week 5, and at 6 weeks after treatment. No differences were observed at weeks 2 and 5 of therapy, but we noted a significant increase in cfDNA in the posttreatment follow-up samples compared with samples collected before CRT (P = .05). Dynamic changes in both cfDNA and ctDNA were observed throughout the course of CRT in patients with locally advanced lung cancer. Specific mutations with therapeutic implications can be identified and tracked using NGS methodologies. Further work is required to characterize the changes in cfDNA and ctDNA over time in patients treated with CRT and to assess the predictive and prognostic potential of this powerful technology.


Inhibiting mild steel corrosion from sulfate-reducing bacteria using antimicrobial-producing biofilms in Three-Mile-Island process water.
Biofilms were used to produce gramicidin S (a cyclic decapeptide) to inhibit corrosion-causing, sulfate-reducing bacteria (SRB). In laboratory studies these biofilms protected mild steel 1010 continuously from corrosion in the aggressive, cooling service water of the AmerGen Three-Mile-Island (TMI) nuclear plant, which was augmented with reference SRB. The growth of both reference SRB (Gram-positive Desulfosporosinus orientis and Gram-negative Desulfovibrio vulgaris) was shown to be inhibited by supernatants of the gramicidin-S-producing bacteria as well as by purified gramicidin S. Electrochemical impedance spectroscopy and mass loss measurements showed that the protective biofilms decreased the corrosion rate of mild steel by 2- to 10-fold when challenged with the natural SRB of the TMI process water supplemented with D. orientis or D. vulgaris. The relative corrosion inhibition efficiency was 50-90% in continuous reactors, compared to a biofilm control which did not produce the antimicrobial gramicidin S. Scanning electron microscope and reactor images also revealed that SRB attack was thwarted by protective biofilms that secrete gramicidin S. A consortium of beneficial bacteria (GGPST consortium, producing gramicidin S and other antimicrobials) also protected the mild steel.


Stigma and psychosocial wellbeing among children affected by parental HIV in China.
Parental HIV infection presents unique psychosocial challenges for families. Affected children are vulnerable to stigma-related distress from a parent's HIV status and are more likely to experience symptoms of depression and low self-esteem than unaffected peers. This study examined whether HIV-related stigma predicted poorer mental health among children affected by parental HIV and whether psychosocial assets mediated this relationship. A sample of 790 children (ages 6-17) affected by parental HIV in Henan, China reported on HIV-related stigma, social support, emotional regulation, future outlook, and mental health symptoms. Structural equation modeling was used to examine the direct and indirect effects of stigma on mental health. Results indicated that stigma was associated with psychosocial resources, which in turn were associated with mental health symptoms. The indirect pathways from stigma to mental health symptoms through psychosocial resources were consistent across age and gender. The experience of HIV-related stigma was associated with poorer overall mental health among children affected by parental HIV in China. Children's psychosocial resources, including social support, emotional regulation, and future outlook, mediated the relationship between HIV-related stigma and mental health symptoms. Future interventions may wish to target these modifiable aspects of wellbeing to improve social and behavioral outcomes in this vulnerable population.


Robust stability of uncertain fractional order systems of neutral type with distributed delays and control input saturation.
Time delay occurs naturally due to the limited bandwidth of any real-world system. However, this problem can deteriorate the system performance and can even result in system instability. Input saturation is also an essential issue due to the energy constraint in real actuators that makes the control design procedure more difficult. This article concerns with the stability of uncertain fractional order (FO) delay systems of neutral type including structured uncertainties, distributed delays and actuator saturation. A Lyapunov-Krasovskii functional allows the formulation of the conditions to insure the asymptotic robust stability of such systems via the linear matrix inequalities (LMI) and to compute the gain of a state feedback controller. In addition, by using the cone complementarity linearization method, we obtain the controller gains that extend the domain of attraction. Several simulations validate the theoretical analysis.


Site-directed mutagenesis of amino acid residues of D1 protein interacting with phosphatidylglycerol affects the function of plastoquinone QB in photosystem II.
Recent X-ray crystallographic analysis of photosystem (PS) II at 1.9-Å resolution identified 20 lipid molecules in the complex, five of which are phosphatidylglycerol (PG). In this study, we mutagenized amino acid residues S232 and N234 of D1, which interact with two of the PG molecules (PG664 and PG694), by site-directed mutagenesis in Synechocystis sp. PCC 6803 to investigate the role of the interaction in PSII. The serine and asparagine residues at positions 232 and 234 from the N-terminus were mutagenized to alanine and aspartic acid, respectively, and a mutant carrying both amino acid substitutions was also produced. Although the obtained mutants, S232A, N234D, and S232AN234D, exhibited normal growth, they showed decreased photosynthetic activities and slower electron transport from QA to QB than the control strain. Thermoluminescence analysis suggested that this slower electron transfer in the mutants was caused by more negative redox potential of QB, but not in those of QA and S2. In addition, the levels of extrinsic proteins, PsbV and PsbU, were decreased in PSII monomer purified from the S232AN234D mutant, while that of Psb28 was increased. In the S232AN234D mutant, the content of PG in PSII was slightly decreased, whereas that of monogalactosyldiacylglycerol was increased compared with the control strain. These results suggest that the interactions of S232 and N234 with PG664 and PG694 are important to maintain the function of QB and to stabilize the binding of extrinsic proteins to PSII.


2-Bromo-1,3-diisopropyl-4,5-dimethyl-1H-imidazol-3-ium dicyanidoargentate.
The title structure, (C(11)H(20)BrN(2))[Ag(CN)(2))], is built up from an approximately C(2v)-symmetric imidazolium cation and a nearly linear dicyanidoargentate anion [N-Ag-N = 176.6 (9)° and Ag-C-N = 178.8 (9) and 177.2 (11)°]. These two constituents are linked by a remarkably short inter-action between the Br atom of the imidazolium cation [C-Br = 1.85 (3) Å] and one N atom of the cyanidoargentate anion [Br⋯N = 2.96 (2) Å], which is much less than the sum of the van der Waals radii (3.40 Å). The crystal studied was twinned by merohedry.


Amino acid changes outside the G-H loop of capsid protein VP1 of type O foot-and-mouth disease virus confer resistance to neutralization by antipeptide G-H serum.
Antiserum to a peptide corresponding to the 135-154 sequence of capsid protein VP1 of the foot-and-mouth disease virus O1 Kaufbeuren was raised in a pig. Although this serum contained neutralizing antibodies, the pig showed clinical symptoms after challenge. Virus isolated from this pig was identified as a mutant, with changes at positions 50, 198 and 211 of VP1 and at position 209 of VP2. This mutant, as well as a plaque isolate of it, differing from the challenge virus at positions 198 on VP1 (alanine being substituted for glutamic acid) and 209 on VP2 (histidine being substituted for tyrosine) resisted neutralization by the anti-peptide serum also in vitro. The same was observed with the O1 Kaufbeuren-related strain O1 Burgwedel, isolated from cattle in the field. It had substitutions only at positions 43 and 101 on VP1. The results show that neutralization epitopes flanking positions 145-147 on VP1 are modulated by other capsid protein parts. These parts seem to be important for neutralization escape in natural FMDV host species.


Highly repetitive DNA families restricted to germ cells in a Japanese hagfish (Eptatretus burgeri): a hierarchical and mosaic structure in eliminated chromosomes.
It is known that in eight hagfishes chromosome elimination occurs during early embryogenesis. The eliminated chromosomes are mostly C-band positive, so that none of the somatic cells have any C-band-positive chromatin. Recently, some highly repetitive DNA sequences have been reported as eliminated elements in these hagfishes based on molecular biological methods. However, no germline-restricted repetitive DNA have been directly isolated from the Japanese hagfish Eptatretus burgeri, from which approximately 21% of the total DNA is eliminated from presumptive somatic cells. Through electrophoretic investigation after digestion with restriction endonucleases, two DNA families that are restricted to germline DNA were isolated. Molecular cloning and sequence analysis revealed that these families are composed of closely related sequences of 64 and 57bp in length, respectively. Southern blot hybridization revealed that the two DNA families are restricted to germline DNA and were thus named EEEb1 and EEEb2, respectively. Moreover, these eliminated elements were highly and tandemly repeated, and it is predicted that they might amplify by saltatory replication and have evolved in a concerted manner. By densitometric scanning, EEEb1 and EEEb2 were found to amount to make up approximately 18.5 and 0.024% of the total germline genomic DNA, accounting for 88.6% of the total eliminated DNA. A fluorescence in situ hybridization experiment demonstrated that EEEb1 is located on all C-band-positive chromosomes that are limited to germ cells, suggesting that EEEb1 is the primary component of eliminated DNA of E. burgeri.


NAD- and NADP-dependent mitochondrially targeted methylenetetrahydrofolate dehydrogenase-cyclohydrolases can rescue mthfd2 null fibroblasts.
Mouse fibroblasts in which the mthfd2 gene encoding mitochondrial NAD-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase (NMDMC) was previously inactivated were infected with retroviral expression constructs of dehydrogenase/cyclohydrolase cDNA. Cellular fractionation confirmed that the expressed proteins were properly targeted to the mitochondria. Expression of the NAD-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase enzyme in mitochondria corrected the glycine auxotrophy of the null mutant cells. A construct in which the cyclohydrolase activity of NMDMC was inactivated by point mutation also rescued the glycine auxotrophy, although poorly. This suggests that the cyclohydrolase activity is also required to ensure optimal production of 10-formyltetrahydrofolate. The expression of the NADP-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase-synthetase in the mitochondria also reversed the glycine requirement of the null cells demonstrating that the use of the NAD cofactor is not absolutely essential to maintain the flux of one-carbon metabolites. All rescued cells demonstrated a decrease in the ratio of incorporation of exogenous formate to serine in standardized radiolabeling studies. This ratio, which is approximately 2.5 for nmdmc(-/-) cells and 0.3 for the wild type cells under the conditions used, is a qualitative indicator of the ability of the mitochondria of the cells to generate formate.


Do work-family initiatives improve employee mental health? Longitudinal evidence from a nationally representative cohort.
Work stress and work-family conflict are important correlates of affective disorders. The article explored (1) whether the wide adoption of work-family initiatives improve a national workforce's mental health; (2) whether the potential benefits differ between the initiatives that give employees autonomy over job quality (flexible schedule and telework) or job quantity (work hours); (3) whether the effects depend on employee's perceived availability or actual usage of the initiatives, and if so, what are the respective mechanisms; and (4) whether there are gender differences in the mental health effects. Fixed-effects analyses of five-wave panel surveys from 2010 to 2020 on a probability sample of 34,484 British workers, which measured mental health with the GHQ-12 scale. Job satisfaction and leisure time satisfaction were tested as mediators. Perceived availability of work-family initiatives improved men and women's mental health by increasing their job satisfaction. Actual usage of work-family initiatives improved women's, but not men's, mental health by increasing their job satisfaction and leisure time satisfaction. The mental health benefits of flexible schedule and telework initiatives are larger than reduced work hours initiatives. The exploratory study used a broad mental health outcome and did not measure work-family initiatives' effects on specific affective disorders such as anxiety and depression. The study could not eliminate time-varying confounders. Actual and perceived job quality are important in workplace mental health promotion. Organizational leaders and policymakers can offer flexible work time and place to reduce work-family conflict and prevent employees' affective disorders.


Beta-cell reactive T-cell clones from type I diabetes patients are not beta cell specific and recognize multiple antigens.
Type I diabetes is the result of an autoimmune destruction of pancreatic beta cells. T cells appear to play a key role in this process. Thus far little information is available on the beta cell antigen or antigens recognized by auto-reactive T cells. Previously, we identified a 38 kD T cell antigen that appears to be localized in the membrane of insulin secretory granules and that is recognized by T cells from newly diagnosed type I diabetes patients. Other groups have reported T cell reactivity against glutamic acid decarboxylase (GAD). To obtain an indication of whether or not beta-cell reactive T cells from type I diabetes patients recognize a limited number of beta-cell antigens, we cloned T-cell lines reactive with rat insulinoma (RIN) membranes from two patients and analysed their antigen specificity. We also studied the antigen specificity of one RIN membrane reactive T-cell clone (1C5), previously isolated from a third patient. From the first patient two identical RIN membrane reactive T-cell clones (7A13) were isolated. The second patient yielded two identical (23A19) RIN membrane reactive T-cell clones, and one that was different (234A33). All clones were CD4+ and saw antigen in the context of different HLA class II alleles. The reactivity of the clones was, however, not restricted to beta cells: all clones showed cross-reactivity with one or more rat tissues, with some preference for those of neuroendocrine origin, but the cross-reactivity patterns were all different. All four clones recognized different fractions electro-eluted from RIN membranes: 29-36 kDa (7A13), 120-170 (23A19), 29-41 (23A33) and 56-72 kDa (1C5). The 23A33 clone reacted with the same 38 kDa fraction electro-eluted from insulinoma membranes as a beta-cell reactive clone (1C6) published previously, but none of the other known beta cell antigen preparations tested were recognized by the T-cell clones. Finally, the subcellular localization of the antigens recognized showed at least two different patterns. These data indicate that beta-cell reactive T cells from the peripheral blood of type I diabetes patients are not necessarily beta-cell specific and may be heterogeneous in regard to their antigen specificity and HLA class II restriction.


Spermatinamine, the first natural product inhibitor of isoprenylcysteine carboxyl methyltransferase, a new cancer target.
Isoprenylcysteine methyltransferase (Icmt) catalyzes the carboxyl methylation of oncogenic proteins in the final step of a series of post-translational modifications. The inhibition of Icmt provides an attractive and novel anticancer target. A natural product high-throughput screening campaign was conducted to discover inhibitors of Icmt. The Australian marine sponge, Pseudoceratina sp., yielded spermatinamine, a novel alkaloid with a bromotyrosyl-spermine-bromotyrosyl sequence, as the bioactive constituent. Its structure was determined by 1D and 2D NMR spectroscopy. Spermatinamine is the first natural product inhibitor of Icmt.


Stroke in Qatar: a first prospective hospital-based study of acute stroke.
Stroke is a major cause of morbidity and mortality in Qatar. The aim of our study was to determine types of strokes, the associated risk factors, clinical presentation, outcome, and time to hospital admission among Qatari and non-Qatari patients as well as young and nonyoung patients. We conducted a hospital-based prospective observational study including all patients admitted to Hamad Medical Corporation with first-ever stroke from September 15, 2004, to September 14, 2005. A stroke was defined according to World Health Organization criteria. Stroke was confirmed in 270 patients of whom 217 (80.4%) had ischemic stroke and 53 (19.6%) had hemorrhagic stroke. Male patients predominated in all types of stroke. The main risk factors for stroke were hypertension and diabetes, whereas lacunar infarct was the most common subtype of ischemic stroke. Risk factor profiles were similar between Qatari and non-Qatari patients except for hypercholesterolemia, which was observed with a higher frequency in Qatari compared with non-Qatari patients with ischemic stroke. There were significant differences between the young and nonyoung patients with ischemic stroke with respect to risk factors, ejection fraction, ventricular wall-motion abnormalities, time to hospital admission, and outcome. Most patients arrived at the hospital more than 3 hours from stroke onset because of unawareness of stroke symptoms. The overall in-hospital mortality was 9.3%. Hypertension and diabetes mellitus were the main risk factors for stroke in Qatar, whereas lacunar infarct was the most common subtype. Significant differences between the young and nonyoung patients were observed with respect to risk factors, ejection fraction, ventricular wall-motion abnormalities, time to hospital admission, and outcome.


Further insights into the regulation of human FAAH by progesterone and leptin implications for endogenous levels of anandamide and apoptosis of immune and neuronal cells.
We have recently reported that leptin (L) and progesterone (P) stimulate the activity and the expression of the endocannabinoid-degrading enzyme anandamide hydrolase (fatty acid amide hydrolase, FAAH) in human lymphoma U937 cells, but not in human neuroblastoma CHP100 cells. We have also shown that leptin and progesterone do not affect the proteins of the endocannabinoid system that synthesize and transport AEA. Here, we have summarized these findings, and have extended them by investigating the effect of leptin and progesterone on the endogenous levels of AEA. We show that leptin and progesterone significantly reduce AEA content in U937 cells (down to approximately 20% and approximately 50% of the controls, respectively), whereas they are ineffective on AEA levels in CHP100 cells. In addition, we show that leptin and progesterone prevent the pro-apoptotic activity of AEA in U937 cells, reducing DNA fragmentation by approximately 50% and approximately 35% compared to controls, respectively. Instead, neither hormone affects apoptosis induced by AEA in CHP100 cells. Since the anti-apoptotic activity of leptin and progesterone parallels their effect on FAAH, it can be suggested that enhanced degradation of AEA is the means to protect U937 cells against the toxicity of this compound. Altogether, these data suggest that a cell-specific regulation of FAAH gene might modulate the apoptotic potential of endocannabinoids along the neuroimmune axis. These findings might be relevant for the development of cell-selective drugs targeted towards FAAH.


Factors influencing enteral autonomy after autologous gastrointestinal reconstructive surgery: A two-centre UK perspective.
The implementation of multidisciplinary care and improvements in parenteral nutrition (PN) in patients with short bowel syndrome (SBS) have led to better outcomes and higher survivability. Autologous gastrointestinal reconstructive (AGIR) surgery can reduce the duration on PN and lead to earlier enteral autonomy (EA). Our aim was to investigate the effect of SBS aetiology and other predictors on the achievement of enteral autonomy following AGIR surgery. Retrospective review of all patients undergoing AGIR surgery in two tertiary paediatric surgical units, between 2010 and 2021. Continuous data is presented as median (range). Twenty-seven patients underwent 29 AGIR procedures (20 serial transverse enteroplasties (STEP), 9 longitudinal intestinal lengthening and tailoring (LILT)) at an age of 6.6 months (1.5 - 104.5). EA rate was 44% at 13.6 months after surgery (1 - 32.8). AGIR procedures achieved an increase in small bowel length of 70% (pre-operative 46.5 vs 77 cm, p = 0.003). No difference was found between STEP and LILT (p = 0.84). Percentage of expected small bowel length (based on the child's weight) was a strong predictor of EA (bowel length >15% - EA 80% vs bowel length ≤15% - EA 17%, p = 0.008). A diagnosis of gastroschisis showed a negative non-significant correlation with the ability to achieve EA (25% vs 60%, p = 0.12). Overall survival rate was 96%. AGIR surgery is an important tool in the multidisciplinary management of children with SBS. Percentage of expected small length and aetiology of SBS are likely predictors of achievement of EA in patients undergoing AGIR surgery. IV: Retrospective Case-Series.


β-Cyclodextrin Assisted Liquid-Liquid Microextraction Based on Solidification of the Floating Organic Droplets Method for Determination of Neonicotinoid Residues.
An efficient and environment-friendly microextraction method, namely, β-cyclodextrin assisted liquid-liquid microextraction, based on solidification of the floating organic droplets method coupled with HPLC is investigated for the sensitive determination of trace neonicotinoid pesticide residues. In this method, β-cyclodextrin is used as a disperser solvent, while 1-octanol is selected as an extraction solvent. β-cyclodextrins was found to decrease interfacial tension and increase the contact area between the organic and water phases with the help of centrifugation. A cloudy solution was rapidly formed and then centrifuged to complete phase separation. Various key parameters influencing extraction efficiency were systematically investigated and optimized; they include salt addition, concentration of β-cyclodextrin, and volume of extraction solvent (1-octanol). Under optimum conditions, good linearity was obtained with coefficient for determination (R2) greater than 0.99. A low limit of detection, high enrichment factor, and good recovery (83 - 132) were achieved. This proves that the proposed method can be applied to determine trace neonicotinoid pesticide residues in natural surface water samples.


Acute Post-Traumatic Aortic Regurgitation.
A 16-year-old girl with a history of blunt chest trauma was admitted because of heart failure symptoms. Transthoracic echocardiography showed severe eccentric aortic regurgitation. Cardiac computed tomography revealed a pseudoaneurysm of the right sinus of Valsalva. We present a rare clinical presentation of a life-threatening condition. (Level of Difficulty: Intermediate.).


Effects of microencapsulation in dairy matrix on the quality characteristics and bioavailability of docosahexaenoic acid astaxanthin.
Compared with free astaxanthin (Asta), docosahexaenoic acid astaxanthin monoester (Asta-C22:6) has higher stability and bioavailability. However, Asta-E is still unable to be used in the water system. Hence it is necessary to build a water-soluble delivery system. In this study, Asta-C22:6 microemulsion and microcapsule using whey protein isolate (WPI) and hydroxypropyl-β-cyclodextrin (HPβ-CD) as composite wall material were prepared. They were added to three dairy products (milk powder, yogurt and flavored dairy product). A dairy product rich in Asta-C22:6 with high bioavailability was designed by measuring quality characteristics, sensory evaluation and in vivo experiments. Compared with spray drying, the freeze-drying microcapsule had a higher encapsulation efficiency (72.5%), water content (4%) and better solubility, and Asta-C22:6 microcapsule (1 g L-1 ) yogurt had the best quality. The bioavailability of Asta-C22:6 microcapsule yogurt was further evaluated. After a single oral dose in mice, the bioavailability of Asta-C22:6 microcapsule in yogurt was significantly increased (Cmax = 0.31 μg mL-1 , AUC0-T = 3.20 h μg mL-1 ). We successfully prepared Asta-C22:6 microcapsule yogurt, which improved the stability and bioavailability of Asta. The present research is meaningful for delivering unstable bioactive small molecules based on WPI and HPβ-CD. It provides an experimental basis for the application of Asta-C22:6 and the development of functional dairy products. © 2022 Society of Chemical Industry.


Cofactor biosynthesis through protein post-translational modification.
Post-translational modifications of amino acids can be used to generate novel cofactors capable of chemistries inaccessible to conventional amino acid side chains. The biosynthesis of these sites often requires one or more enzyme or protein accessory factors, the functions of which are quite diverse and often difficult to isolate in cases where multiple enzymes are involved. Herein is described the current knowledge of the biosynthesis of urease and nitrile hydratase metal centers, pyrroloquinoline quinone, hypusine, and tryptophan tryptophylquinone cofactors along with the most recent work elucidating the functions of individual accessory factors in these systems. These examples showcase the breadth and diversity of this continually expanding field.


To Sandwich Technetium: Highly Functionalized Bis-Arene Complexes [99m Tc(η6 -arene)2 ]+ Directly from Water and [99m TcO4 ].
The labeling of (bio)molecules with metallic radionuclides such as 99m Tc demands conjugated, multidentate chelators. However, this is not always necessary since phenyl rings can directly serve as integrated, organometallic ligands. Bis-arene sandwich complexes are generally prepared by the Fischer-Hafner reaction. In extension of this, we show that [99m Tc(η6 -C6 R6 )2 ]+ -type complexes are directly accessible from water and [99m TcO4 ]- , even using arenes incompatible with Fischer-Hafner conditions. To unambiguously confirm the nature of these unprecedented 99m Tc complexes, their rhenium homologous have been prepared by substituting naphthalene ligands in [Re(η6 -C10 H8 )2 ]+ with the corresponding phenyl groups. The ease with which highly stable [99m Tc(η6 -C6 R6 )2 ]+ complexes are formed under standard labeling conditions enables a multitude of new potential imaging agents based on commercial pharmaceuticals or lead structures.


Health outcomes with catheter ablation or antiarrhythmic drug therapy in atrial fibrillation: results of a propensity-matched analysis.
Patients with atrial fibrillation (AF) face significant risks of stroke and heart failure. The objective of this study was to determine whether AF ablation reduces the long-term risk of stroke or heart failure compared with antiarrhythmic drug therapy. A coding algorithm was used to identify AF patients treated with catheter ablation (n=3194) or antiarrhythmic drugs without ablation (n=6028) between 2005 and 2009 using The MarketScan Research Database from Thomson Reuters From this sample, 801 pairs were propensity matched, based on 15 demographic and clinical characteristics and baseline medication use. Rates of stroke/transient ischemic attack (TIA) and heart failure hospitalizations for up to 3 years were examined. Patients treated with catheter ablation had a significantly lower rate of stroke or TIA (3.4% per year) than a group of patients with AF managed with antiarrhythmic drugs only (5.5% per year), with an unadjusted hazard ratio of 0.62 (95% CI, 0.44-0.86; P=0.005). The rates for heart failure hospitalization were 1.5% per year in the ablation group and 2.2% per year in the antiarrhythmic drug group, with an unadjusted hazard ratio of 0.69 (95% CI, 0.42-1.15; P=0.158). These results were minimally altered in Cox proportional hazards models, which further adjusted for potential confounders not well balanced by the propensity matching. In a large propensity-matched community sample, AF ablation was associated with a reduced risk of stroke/TIA and no significant difference in heart failure hospitalizations compared with antiarrhythmic drug therapy. These findings require confirmation with randomized study designs.


Isolation and characterization of 13 microsatellites for the rare endemic shrub Tetratheca erubescens (Elaeocarpaceae).
Microsatellite markers were developed for the rare Tetratheca erubescens (Elaeocarpaceae) to assess genetic diversity and spatial structuring. We generated ca. 2.7 million sequence reads using a Personal Genome Machine (PGM) semiconductor sequencer. Using the QDD pipeline, we designed primers for >12,000 sequences with PCR product lengths of 80-480 bp. From these, 30 primer pairs were selected and screened using PCR, from which 11 loci were found to be polymorphic and amplified reliably. For a sample of 95 plants from three populations, the number of alleles observed for these 11 loci ranged from two to seven and expected heterozygosity ranged from 0.06 to 0.72. No consistent evidence for null alleles or departure from Hardy-Weinberg equilibrium was found for any of the 11 loci. These markers will enable the quantification of genetic impact of proposed mining activities on the narrow endemic T. erubescens.


Prognostic significance of angina pectoris before first acute myocardial infarction.
To delineate the clinical significance and prognostic importance of a history of chronic or new onset angina pectoris before acute myocardial infarction (AMI), 732 consecutive patients admitted for a first AMI were studied and divided into 3 groups. Two hundred patients (27%) had chronic angina before AMI (greater than 1 month); 247 patients (34%) had new onset angina before AMI (less than 1 month) and the 285 remaining patients (39%) never had angina before AMI. All clinical characteristics were similar in the group of patients with chronic angina and in the group of patients with new onset angina, including in-hospital mortality (10 vs 9%) and 3-year post-hospital mortality (16 vs 16%). Compared to the 285 patients without angina, the 447 patients with angina before AMI were older, more likely to be women, and had a higher frequency of anterior AMI and early post-infarction angina. Both groups had a similar in-hospital mortality (10 vs 8%, not significant), but patients with angina had a higher 3-year post-hospital mortality (16 vs 7%, p less than 0.001). In the group of patients with angina before AMI who were discharged from the hospital, the comparison of nonsurvivors and survivors showed that the patients who died were older, presented more frequently with a non-Q-wave myocardial infarct and more often had left ventricular failure and complete bundle branch block during hospital stay. Chronic and new onset angina before AMI have the same clinical characteristics and deleterious long-term prognostic significance.


Incubation time for AIDS from French transfusion-associated cases.
Although incubation time is a key parameter of the epidemiology of AIDS, statistical estimates based on transfusion-associated AIDS cases have, up to now, used only the single dataset provided by the AIDS program of the Centers for Disease Control (CDC) in Atlanta. Using a new dataset provided by the Direction Générale de la Santé (DGS), of the French Ministry of Health1, we estimate the mean incubation time for AIDS (median in brackets) to be 5.3 years (5.3 years) with a 90% confidence interval ranging from 4.4 to 8.9 years (4.4 to 8.8 years), when a Weibull distribution is postulated for incubation time. The previously encountered problem of very large confidence intervals (range larger than 100 years), is not observed, indicating that an accurate estimate for mean incubation time will be obtainable in the near future.


The resting metabolic rate of people with severe mental illness: a systematic review and meta-analysis.
People with severe mental illness (SMI), including schizophrenia and related psychoses and bipolar disorder, are at greater risk for obesity compared with people without mental illness. An altered resting metabolic rate (RMR) may be a key driving factor; however, published studies have not been systematically reviewed. This systematic review and meta-analysis aimed to determine whether the RMR of people with SMI assessed by indirect calorimetry differs from (i) controls, (ii) predictive equations and (iii) after administration of antipsychotic medications. Five databases were searched from database inception to March 2022. Thirteen studies providing nineteen relevant datasets were included. Study quality was mixed (62 % considered low quality). In the primary analysis, RMR in people with SMI did not differ from matched controls (n 2, standardised mean difference (SMD) = 0·58, 95 % CI -1·01, 2·16, P = 0·48, I2 = 92 %). Most predictive equations overestimated RMR. The Mifflin-St. Jeor equation appeared to be most accurate (n 5, SMD = -0·29, 95 % CI -0·73, 0·14, P = 0·19, I2 = 85 %). There were no significant changes in RMR after antipsychotic administration (n 4, SMD = 0·17, 95 % CI -0·21, 0·55, P = 0·38, I2 = 0 %). There is little evidence to suggest there is a difference in RMR between people with SMI and people without when matched for age, sex, BMI and body mass, or that commencement of antipsychotic medication alters RMR.


Ethidium tetra-phenyl-borate aceto-nitrile disolvate.
In the title solvated salt, (C21H20N3){B(C6H5)4}·2CH3CN (systematic name 3,8-di-amino-5-ethyl-6-phenyl-phenanthridin-5-ium tetra-phenyl-borate aceto-nitrile disolvate), the dihedral angle between the tricyclic fused ring system (r.m.s. deviation = 0.021 Å) and the pendant phenyl group of the ethidium cation is 84.91 (7)°. The {B(C6H5)4}- anion has a typical tetra-hedral structure. The aceto-nitrile solvent mol-ecules do not accept hydrogen bonds from the -NH2 groups of the ethidium ions.


Characteristics of referral to a multidisciplinary palliative home care team.
Until now, in Belgium at least, no scientific information has been available about which patients are referred to multidisciplinary palliative home care teams (MHCT) and by which caregivers, nor about timing of referral and factors associated with it. A prospective cohort study was carried out during a nine-month period in the oldest Flemish MHCT, and included all patients for whom a palliative care trajectory had been initiated. In total, 325 patients (54% male, mean age 71 years, 83% cancer patients) were referred: 25% by their GP, 24% by hospital social workers, and 22% by the patient's family. Median timing of referral was 26 days (IQR 8-78) before death. Patients of 75 years or older were more likely to have been referred later by comparison with younger patients. Timely referral to a MHCT seems to be difficult and not equal for all patients. Referral timing varies largely by age groups and diseases.


Ejaculatory status and fertility rates after primary retroperitoneal lymph node dissection.
We determined the incidence of antegrade emission after primary retroperitoneal lymph node dissection in a large contemporary cohort. Our secondary purpose was to evaluate the fertility rate in this population. We queried the testicular cancer database at our institution from January 1, 2000 to December 31, 2005 and identified all 280 patients who underwent primary retroperitoneal lymph node dissection. Of these patients we contacted 176, and questioned them about ejaculatory and fertility status at 3 to 9 years of followup. Of 176 patients who underwent primary retroperitoneal lymph node dissection 171 (97%) reported preserved antegrade emission. Of the 135 men who underwent a nerve sparing procedure 134 (99%) could ejaculate, as could 33 of 37 (89%) who underwent nonnerve sparing surgery. An attempt to father children was reported by 64 men, of whom 47 (73.4%) were successful. Three other patients fathered children via in vitro fertilization. Most men who undergo modern primary retroperitoneal lymph node dissection maintain antegrade emission and ejaculation.


Acrylamide causes neurotoxicity by inhibiting glycolysis and causing the accumulation of carbonyl compounds in BV2 microglial cells.
Acrylamide is a potent neurotoxin produced during industrial production or food processing at high temperatures, and its effect on glycolytic processes may be critical in triggering neurotoxicity. Our work investigated the neurotoxic effects of acrylamide based on glycolysis using immortalized mouse microglia cell line BV2. The results showed that 1.5 mM acrylamide significantly inhibited the expression and activity of triphosphate isomerase and 3-phosphoglyceraldehyde dehydrogenase. Moreover, acrylamide limited the expression of pyruvate kinase and the decrease of pyruvate and lactate content of glycolysis products. In addition, acrylamide could increase intracellular methylglyoxal content and further affect its detoxification system. Not only that, subsequent cellular responses resulting from methylglyoxal accumulation, such as oxidative stress, activation of ERK, upregulation of NF-κB inflammatory signaling pathway, and elevated pro-inflammatory factor TNF-α, were found in the acrylamide-treated cell model. Therefore, we suggest that acrylamide's perturbation of the glycolytic process leads to methylglyoxal accumulation, which may be responsible for its key to neurotoxicity.


Electric birefringence of myosin subfragments.
Electric birefringence measurements have been made on aqueous solutions of myosin subfragments, heavy meromyosin, subfragments 1 and 2 (S-1 and S-2). All of these showed positive electric birefringence. Heavy meromyosin and S-2 showed a large intrinsic Kerr constant. From the analysis of the build up and decay process of the birefringence, the contribution of the slow induced dipole moment was concluded in heavy meromyosin and S-2, although the existence of the permanent dipole moment was not completely excluded. The decay process of the birefringence of heavy meromyosin was found to consist of two components; the fast one of which had a relaxation time of the same order as that of S-1. This is probably due to the presence of a flexible hinge in heavy meromyosin.


Polygenic risk score for problematic alcohol use predicts heavy drinking and alcohol use disorder symptoms in young adulthood after accounting for adolescent alcohol use and parental alcohol use disorder.
Genome-wide association studies (GWAS) have identified hundreds of loci associated with alcohol-related traits. GWAS permit the calculation of polygenic risk scores (PRS), which aggregate genetic risk for a trait across the genome. To evaluate the usefulness of a PRS for problematic alcohol use (PAU)-which subsumes alcohol use disorder (AUD) and alcohol-related problems-we tested whether this PRS predicted heavy drinking and alcohol problems after accounting for family history of AUD and prior drinking history, robust and established predictors of PAU. Participants (N=665) were European-ancestry members of the Michigan Longitudinal Study, a prospective family study with high rates (65%) of parental AUD. Participants reported their frequency of alcohol use, maximum drinks consumed in a 24-hour period, and alcohol use problems at four assessments in adolescence and young adulthood (11-29 years old). We used polygenic prediction via Bayesian regression and continuous shrinkage priors to create a PAU PRS using summary statistics from a meta-GWAS of PAU. After adjusting for demographic covariates, parental AUD, and drinking and alcohol use problems in early and mid/late adolescence, the PAU PRS was significantly associated with alcohol-related problems in young adulthood (β=.08, p=.047; R2=0.6%). The PAU PRS also had a significant indirect effect on alcohol use problems in young adulthood through earlier drinking and alcohol use problems (β=.02, p=.03). The PAU PRS predicted alcohol problems in young adulthood after accounting for parental history of AUD and alcohol use in adolescence, providing evidence that genetic data uniquely inform the etiology of alcohol problems.


The effect of tetrathiomolybdate upon sheep caeruloplasmin amine oxidase activity in vitro: the influence of substrate on apparent sensitivity to inhibition.
The effect of molybdate, dithiomolybdate, tetrathiomolybdate and diethyldithiocarbamate on caeruloplasmin amine oxidase activity was examined using o-dianisidine as a substrate. Enzyme activity was strongly inhibited by tetrathiomolybdate (1.5 X 10(6)M), oxidation of another substrate, p-phenylenediamine, was inhibited only slightly.


Influence of dopamine infusion on plasma prolactin released by kidney capsule transplanted anterior pituitaries.
The infusion of dopamine into the renal artery resulted in decreased prolactin release from 3 anterior pituitary glands transplanted under the kidney capsule. Prolactin levels continually decreased over a 5 min period after DA infusion was terminated and thereafter approached preinfusion levels by the end of 10 min.


Influence of potassium thiocyanate on thyroid function of rainbow trout, Salmo gairdneri.
Sublethal doses of potassium thiocyanate (KSCN), an iodide competitor, depressed thyroidal radioiodide uptake of fed laboratory rainbow trout at 11 degrees, but did not depress either plasma levels of T4 (L-thyroxine) and T3 (triiodo-L-thyronine) or radioiodide incorporation into plasma iodothyronines. Furthermore, plasma T4 levels of KSCN-treated trout increased to levels comparable to those of control trout in response to bovine TSH. The apparent inability of KSCN to inhibit thyroid hormone synthesis and release may be due to non-carrier-mediated iodide diffusion into the thyroid from the high iodide concentration normally found in plasma of freshwater salmonids. In conclusion, KSCN is not a practical inhibitor of thyroid hormone release in laboratory trout, and thyroid radioiodide uptake is a highly misleading index of thyroidal status.


Multi-Layer π-Stacked Molecules as Efficient Thermally Activated Delayed Fluorescence Emitters.
Multi-layer π-stacked emitters based on spatially confined donor/acceptor/donor (D/A/D) patterns have been developed to achieve high-efficiency thermally activated delayed fluorescence (TADF). In this case, dual donor moieties and a single acceptor moiety are introduced to form two three-dimensional (3D) emitters, DM-BD1 and DM-BD2, which rely on spatial charge transfer (CT). Owing to the enforced face-to-face D/A/D pattern, effective CT interactions are realized, which lead to high photoluminescence quantum yields (PLQYs) of 94.2 % and 92.8 % for the two molecules, respectively. The resulting emitters exhibit small singlet-triplet energy splitting (ΔEST ) and fast reverse intersystem crossing (RISC) processes. Maximum external quantum efficiencies (EQEs) of 28.0 % and 26.6 % were realized for devices based on DM-BD1 and DM-BD2, respectively, which are higher than those of their D/A-type analogues.


Nature-inspired design of motif-specific antibody scaffolds.
Aberrant changes in post-translational modifications (PTMs) such as phosphate groups underlie a majority of human diseases. However, detection and quantification of PTMs for diagnostic or biomarker applications often require PTM-specific monoclonal antibodies (mAbs), which are challenging to generate using traditional antibody-selection methods. Here we outline a general strategy for producing synthetic, PTM-specific mAbs by engineering a motif-specific 'hot spot' into an antibody scaffold. Inspired by a natural phosphate-binding motif, we designed and selected mAb scaffolds with hot spots specific for phosphoserine, phosphothreonine or phosphotyrosine. Crystal structures of the phospho-specific mAbs revealed two distinct modes of phosphoresidue recognition. Our data suggest that each hot spot functions independently of the surrounding scaffold, as phage display antibody libraries using these scaffolds yielded >50 phospho- and target-specific mAbs against 70% of target peptides. Our motif-specific scaffold strategy may provide a general solution for rapid, robust development of anti-PTM mAbs for signaling, diagnostic and therapeutic applications.


The influence of adjuvant therapy on cardiorespiratory fitness in early-stage breast cancer seven years after diagnosis: the Cooper Center Longitudinal Study.
We examined cardiorespiratory fitness (CRF) levels in early stage breast cancer patients and determined whether CRF differs as a function of adjuvant therapy regimen. A total of 180 early breast cancer patients representing three treatment groups (surgery only, single-, and multi-modality adjuvant therapy) in the Cooper Center Longitudinal Study (CCLS) were studied. A non-cancer control group (n = 180) matched by sex, age, and date of the CCLS visit was included. All subjects underwent an incremental exercise tolerance test to symptom limitation to assess CRF (i.e., peak metabolic equivalents [METs] and time to exhaustion). The mean time from breast cancer diagnosis to exercise tolerance testing was 7.4 ± 6.2 years. In adjusted analyses, time to exhaustion and peak METs were incrementally impaired with the addition of surgery, single-, and multi-modality adjuvant therapy compared to those of matched controls (p = 0.006 and 0.028, respectively). CRF was lowest in the multi-modality group compared to all other groups (all p's < 0.05). Despite being 7 years post-diagnosis, asymptomatic early breast cancer survivors have marked reductions in CRF. Patients treated with multi-modal adjuvant therapy have the greatest impairment in CRF.


[Hemodynamic monitoring of critically ill patients : Bedside integration of data].
Hemodynamic monitoring of critically ill patients is a key issue in intensive care medicine. Indication and application of invasive hemodynamic monitoring is a highly complex matter and requires thorough professional education and training. A literature review was performed. A pragmatic approach can be divided into several steps such as medical history, physical examination, imaging, and laboratory results, which support the primary working diagnosis and allow further clarification of the underlying pathophysiology. Invasive arterial blood pressure and cardiac output measurement as well as components of the functional hemodynamic monitoring help to assess fluid responsiveness and to guide volume loading, diuretic therapy as well as administration of vasoactive or positive inotrope substances. All information gathered through medical history, physical examination, imaging, and hemodynamic monitoring help to form an overall picture and should be reevaluated regularly and in individual cases very closely depending on the hemodynamic instability of the patient. Target values are strictly indicative and are not binding taking into account that each patient has its unique pathophysiological profile.


A new modelling approach for predicting process evolution of cork-rubber composites slabs vulcanization.
In order to predict the evolution of the vulcanization process of cork-rubber slabs, a numerical approach was developed combining heat transfer by conduction and kinetics models. A one-dimensional model was applied to predict the evolution of temperature and degree of cure at different stages of the vulcanization of a cork-rubber composite. Also, due to the degradation verified by the compound, an existent reversion model was added to the problem. Based on rheometer data, cure and reversion parameters were determined. Experimental data were used to determine the thermal properties of the compound, assuming a constant value or according to its degree of cure and temperature. The results obtained by simulation showed a good correspondence with experimental results, even when assuming constant thermal properties. The application of the proposed methodology provides information about the optimum process parameters for each thickness slab, without compromising the homogeneity and characteristics of the final product, which can be a valuable tool during the development and product stages of cork-rubber composites.


[Synthesis of periodic mesoporous ethanesilica and its application in high performance liquid chromatography].
The periodic mesoporous ethanesilica (PME) was synthesized in acidic medium by the condensation of 1,2- bis (trimethoxysily) ethane (BTME) using triblock copolymer EO20PO70EO20 (P123) as template, cetyltrimethylammonium bromide (CTAB) as co-surfactant and ethanol as co-solvent. The results of nitrogen-sorption measurement, powder X-ray diffraction (XRD), and scanning electron microscopy (SEM) show that the PME has high surface area (1152 m2/g), narrow pore-size distribution, ordered pore structure, and spherical morphology. The spherical PME without further chemical modification was used as the stationary phase in reversed-phase high performance liquid chromatography. Highly efficient and rapid separation of the mixture of polycyclic aromatic hydrocarbons (benzene, naphthalene, biphenyl, phenanthrene and pyrene) was obtained on the column packed with the PME.


Language network self-inhibition and semantic similarity in first-episode schizophrenia: A computational-linguistic and effective connectivity approach.
A central feature of schizophrenia is the disorganization and impoverishment of language. Recently, we observed higher semantic similarity in first-episode-schizophrenia (FES) patients. In this study, we investigate if this aberrant similarity relates to the 'causal' connectivity between two key nodes of the word production system: inferior frontal gyrus (IFG) and the semantic-hub at the ventral anterior temporal lobe (vATL). Resting-state fMRI scans were collected from 60 participants (30 untreated FES and 30 healthy controls). The semantic distance was measured with the CoVec semantic tool based on GloVe. A spectral dynamic causal model with Parametrical Empirical Bayes was constructed modelling the intrinsic self-inhibitory and extrinsic-excitatory connections within the brain regions. We estimated the parameters of a fully connected model with the semantic distance as a covariate. FES patients chose words with higher semantic similarity when describing the pictures compared to the HC group. Among patients, an increased semantic similarity was related with an increase in intrinsic connections within both the vATL and IFG, suggesting that reduced 'synaptic gain' in these regions likely contribute to aberrant sampling of the semantic space during discourse in schizophrenia. Lexical impoverishment relates to increased self-inhibition in both the IFG and vATL. The associated reduction in synaptic gain may relate to reduced precision of locally generated neural activity, forcing the choice of words that are already 'activated' in a lexical network. One approach to improve word sampling may be via promoting synaptic gain via supra-physiological stimulation within the Broca's-vATL network; this proposal needs verification.


Efficacy and Safety Profile of Diclofenac/Cyclodextrin and Progesterone/Cyclodextrin Formulations: A Review of the Literature Data.
According to health technology assessment, patients deserve the best medicine. The development of drugs associated with solubility enhancers, such as cyclodextrins, represents a measure taken in order to improve the management of patients. Different drugs, such as estradiol, testosterone, dexamethasone, opioids, non-steroidal anti-inflammatories (NSAIDs; i.e. diclofenac), and progesterone are associated with cyclodextrins. Products containing the association of diclofenac/cyclodextrins are available for subcutaneous, intramuscular, and intravenous administration in doses that range from 25 to 75 mg. Medicinal products containing the association of progesterone/cyclodextrins are indicated for intramuscular and subcutaneous injection at a dose equal to 25 mg. The effects of cyclodextrins have been discussed in the solubility profile and permeability through biological membranes of drug molecules. A literature search was performed in order to give an overview of the pharmacokinetic characteristics, and efficacy and safety profiles of diclofenac/hydroxypropyl-β-cyclodextrin (HPβCD) and progesterone/HPβCD associations. The results of more than 20 clinical studies were reviewed. It was suggested that the new diclofenac/HPβCD formulation gives a rapid and effective response to acute pain and, furthermore, has pharmacokinetic and efficacy/safety profiles comparable to other medicinal products not containing cyclodextrins. One of the principal aspects of these new diclofenac formulations is that in lowering the dose (lower than 50 mg) the drugs could be more tolerable, especially in patients with comorbid conditions. Moreover, results of studies investigating the characteristics of progesterone and cyclodextrins showed that the new formulation (progesterone/HPβCD 25 mg solution) has the same bioavailability as other products containing progesterone. It is more rapidly absorbed and allows the achievement of peak plasma concentrations in a shorter time. Finally, the new formulation of progesterone was shown to be safe and not inferior to other products already on the market, with the exception of progesterone administered vaginally. As shown by the results of clinical studies presented in this review, the newly approved medicines containing cyclodextrins have been found to be as effective and as well-tolerated as other medicinal products that do not contain cyclodextrins. Moreover, the newly approved lower dose of diclofenac associated with cyclodextrins is consistent with the European Medicines Agency recommendations reported in the revision of the Assessment Report for Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) and Cardiovascular Risk. Finally, the use of cyclodextrins led to significant increases in solubility and bioavailability of drugs, such as diclofenac and progesterone, and improvement in the efficacy and safety of these drugs.


Dimethylbutyrylated Dibenzocyclooctadiene Lignans from the Fruits of Schisandra cauliflora Inhibit NO Production in LPS-Activated RAW264.7 Cells.
From the fruits of Schisandra cauliflora, five new dimethylbutyrylated dibenzocyclooctadiene lignans, named schisandracaurins A-E, were isolated using separation and chromatographic techniques. Their structures were determined by extensive analyses of HR-ESI-MS, NMR, and ECD spectra. The schisandracaurins A-E potentially inhibited NO production in LPS-activated RAW264.7 cells with their IC50 values from 21.4 to 30.3 μM.


Different pattern of carotid and myocardial changes according to left ventricular geometry in hypertensive patients.
The relation between left ventricular (LV) hypertrophy and LV function is well known. However, less is known about the vascular changes influenced by LV geometry. We sought to investigate the relationship of LV geometry to carotid arterial and LV function. A total of 476 hypertensive patients were prospectively recruited. All subjects underwent echocardiography and carotid ultrasound. LV geometry is categorized into four groups according to relative wall thickness (RWT) and LV mass index (LVMI). Concentric LV geometry was associated with increased carotid intima-media thickness (IMT), β-stiffness, and lower strain. All of the carotid parameters showed a stepwise change according to RWT of LV, whereas LV function was worse in hypertrophic geometry, as reflected by significantly lower systolic mitral annular velocity, higher left atrial volume index and E/E' ratio (P<0.001). By multivariate analysis after adjustment for clinical and laboratory parameters, IMT was independently associated with RWT, whereas myocardial function was independently associated with LVMI. Carotid arterial function and IMT showed worse values in concentric geometry, whereas LV systolic and diastolic function were worse in hypertrophic geometry, suggesting a discrepancy between carotid arterial and LV function in hypertensive patients.


The role of brain natriuretic peptide in systolic heart failure.
Heart failure (HF) is a progressive multisystem disease that involves neurohormonal activation, dysfunction of cardiac and skeletal musculature, and a host of other pathological changes. The neurohormonal activation in HF triggers the release of the natriuretic peptides. One peptide of particular interest is brain natriuretic peptide (BNP). It is primarily released by the ventricles of the heart and has adaptive function in counteracting the effects of neurohormonal activation in patients with HF. The focus of this article is the discussion of the physiology of BNP as well as its role in systolic HF, although it also plays a role in diastolic HF.


The phenotypic diversity in the production performance of Alabio ducks (Anas platyrhynchos Borneo) in South Kalimantan, Indonesia.
This study aimed to examine the diversity of phenotypic characteristics of female Alabio ducks (Anas platyrhynchos Borneo) and their eggs in South Kalimantan, Indonesia. A total of 200 18-month-old ducks and 300 eggs were selected using a survey method with multistage random sampling. These samples were obtained from two districts in the province of South Kalimantan [Banjar (BJ) districts and Tanah Laut (TL)]. The observed data were analyzed descriptively with variations using the independent t-test for each location. Principal component analysis (PCA) was deployed to assess the overall variance and define variables with greater discriminatory power between individuals. The observations on the body's physical characteristics and the eggs of Alabio ducks showed significant differences between ducks from BJ and TL areas (p < 0.05) except for yolk diameter, albumen height, albumen index, and Haugh unit (HU) (p > 0.05). Most egg quality traits from BJ were greater than those from TL, which include egg weight, egg length, egg width, yolk weight, albumen weight, shell weight, shell thickness, and egg shape index. The PCA revealed medium-to-high communalities in the phenotypic body characteristics of Alabio ducks and their eggs. Significant physical characteristics and egg quality differences were found between the two locations, except for the albumen index and HU, with substantial variability within each trait. These traits could explain the total variation in the phenotypic characteristics of female Alabio ducks.


Genome-wide identification of the HDAC family proteins and functional characterization of CsHD2C, a HD2-type histone deacetylase gene in tea plant (Camellia sinensis L. O. Kuntze).
The histone deacetylases (HDACs) are involved in growth, development and stress responses in many plants. However, the functions of HDACs in tea plant (Camellia sinensis L. O. Kuntze) and other woody plants remain unclear. Here, 18 CsHDAC genes were identified by genome-wide analysis in tea plant. The phylogenetic analysis demonstrated that the CsHDAC proteins were divided into three subfamilies, namely, the RPD3/HDA1 subfamily (8 members), the SIR2 subfamily (4 members) and the plant specific HD2 subfamily (6 members). The expression patterns showed that most members of CsHDACs family were regulated by different abiotic stress. High correlation was found between the expression of the CsHDACs and the accumulation of theanine, catechin, EGCG and other metabolites in tea plant. Most of the CsHDAC proteins were negative regulators. We further studied a specific gene CsHD2C (NCBI-ID: KY364373) in tea plant, which is the homolog of AtHD2C, encoded a protein of 306 aa. CsHD2C was highly expressed in leaves, young buds and stems. The transcription of CsHD2C was inhibited by ABA, NaCl and low temperature. It was found localized in the nucleus when fused with a YFP reporter gene. Overexpression of CsHD2C can rescue the phenotype related to different abiotic stresses in the mutant of AtHD2C in Arabidopsis. The stress-responsive genes RD29A, RD29B, ABI1 and ABI2 were also investigated to understand the regulating role of CsHD2C under abiotic stresses. We also found that CsHD2C could renew the change of acetylation level for histone H4 and the RNAP-II occupancy accumulation in the promoter of abiotic stress responses gene in the hd2c Arabidopsis mutant. Together, our results suggested that CsHD2C may act as a positive regulator in abiotic stress responses in tea plant.


Effect of incubating human sperm at room temperature on capacitation-related events.
To determine the effect of human sperm incubation at room temperature (20 degrees C) upon capacitation-related events. Prospective study. Basic research laboratory. Semen samples were obtained from normozoospermic volunteers. Human follicular fluid (hFF) was collected from women undergoing assisted reproductive treatment. Spermatozoa were incubated for up to 18 hours at 20 degrees C and/or 37 degrees C. Protein tyrosine phosphorylation patterns, development of hyperactivated motility, and induction of acrosome reaction (AR) in response to hFF. Spermatozoa incubated for 18 hours at 20 degrees C showed an array of tyrosine phosphorylated proteins similar to noncapacitated cells. After incubation at 20 degrees C, the percentage of spermatozoa displaying hyperactivated motility and undergoing acrosomal loss in response to hFF was significantly lower when compared with cells kept the same time at 37 degrees C. Conversely, spermatozoa incubated overnight at 37 degrees C could respond to hFF, either at 37 degrees C or 20 degrees C. When preincubation at 20 degrees C was followed by sperm exposure to 37 degrees C, capacitation-related events could be activated. In capacitated cells (16 hours at 37 degrees C), 2-hour incubation at 20 degrees C led to a significant decrease in acrosome reaction inducibility, suggesting sperm decapacitation. Human sperm incubation at room temperature does not allow capacitation, although it does not affect hFF-induced acrosome reaction in capacitated cells. The blocking effect is overcome when spermatozoa are exposed to 37 degrees C.


Management of cutaneous malignant melanoma by dermatologists of the American Academy of Dermatology. II. Definitive surgery for malignant melanoma.
During the past few decades there has been increasing interest and training in dermatologic surgery. Our purpose was to determine to what extent members of the American Academy of Dermatology (AAD) are involved in the surgical management of patients with malignant melanomas (MMs), comparing 1982 with 1992. Members of the AAD practicing in the United States (N = 7412) were sent a questionnaire that surveyed their role in the definitive treatment of patients with MMs and the surgical margins of normal-appearing skin that they used or recommended for melanomas of various thicknesses. Sixty-four percent of the respondents stated that they performed the definitive surgery for in situ melanoma in 1992, a 14% increase from 1982. Although a significantly greater percentage of dermatologists were performing the definitive surgery for invasive melanoma in 1992 (28%) compared with 1982 (14%), the majority continued to refer their patients to surgical colleagues for definitive treatment. There has been a narrowing of surgical margins recommended or used for melanomas of all thicknesses. In addition, regional differences of the role of the dermatologist in surgical management of patients with MM were observed. An increasing proportion of dermatologists are involved in the surgical management of patients with MMs. Most dermatologists appear to be in accord with the guidelines for surgical margins currently recommended in the literature.


Carotid Endarterectomy in the Southern California Vascular Outcomes Improvement Collaborative.
The objective of this study was to examine the variation in practice patterns and associated outcomes for carotid endarterectomy (CEA) within the Southern California Vascular Outcomes Improvement Collaborative (So Cal VOICe), a regional quality group of the Vascular Quality Initiative. All cases entered in the CEA registry by the So Cal VOICe were included in the study. From September 2010 through September 2015, 1,110 CEA cases were entered by 9 centers in the So Cal VOICe. Six hundred seventy-seven patients (61%) were male with mean age of 73 years. Nine hundred eighty-eight (89%) were hypertensive, 655 (59%) were prior or current smokers, 389 (35%) were diabetics, and 233 (21%) had coronary artery disease. Eight hundred twenty-one (74%) patients were asymptomatic (no history of ipsilateral neurologic event). The percentage of asymptomatic patients varied across the 9 centers from 57% to 91%. Preoperatively, 344 (31%) underwent cardiac stress test, center variation 13-75%, 500 (45%) underwent only duplex, center variation 11-72%. Intraoperatively, 600 (54%) underwent routine shunting, whereas 67 (6%) were shunted for an indication, and 444 (40%) were not shunted. Wound drainage was used in 422 (38%) cases, center variation 2-98%. Completion imaging by duplex and/or angiogram was performed in 766 (69%) cases, center variation 0-100%. Postoperatively, 11 (1%) patients had a new ipsilateral postoperative neurologic event, center variation 0-1.3%, 6 (0.5%) had a postoperative myocardial infarction, center variation 0-1.3%, and 8 (0.7%) required return to operating room for bleeding, center variation 0-1.3%. Despite wide variation in practice patterns surrounding CEA in the So Cal VOICe, postoperative complications were uniformly low. Further work will focus on identifying practices that can be modified to improve cost-effectiveness while maintaining excellent outcomes.


Rhythmic synthesis of various 5-methoxyindoles in the pineal gland of male adult golden hamsters, kept under the same artificial conditions throughout the year.
Until now the day/night and seasonal rhythmicity in the synthesis of 5-methoxyindoles (MI) is thought to be regulated by environmental factors, especially photoperiod and temperature. Endogenous factors are also implicated in the generation of N-acetyltransferase and hydroxyindole-O-methyltransferase activity rhythms. In the present experiments seasonal rhythmicity in the synthesis of MI in the pineal gland was investigated in hamsters kept under the same artificial conditions throughout the year. Though the environmental conditions were the same, day/night and seasonal rhythmicity in the production of MI in the pineal were observed indicating the existence of endogenous factors influencing the rhythmicities. In November, most of the MI showed the highest synthesis, MA and ML excepted, which were especially produced in July and September. The results obtained sustain the hypothesis that aMT is synthesized from MT rather than from aHT. Moreover, the rhythmicities in aMT synthesis are not identical to those found in aMT concentration as described in the literature. This indicates that synthesis and concentration of a compound are not comparable. At the end of the light period, when aMT injections have an antigonadotropic effect, a peak of aMT synthesis was always present. Although MI synthesis showed seasonal rhythmicity, no reproductive cycle occurred in the hamsters. At present, the concept that the pro- and/or antigonadal effects of the pineal are mediated by aMT seems to be the most acceptable. The present results, however, indicate that aMT and perhaps other MI, often regarded as factors influencing gonadal growth in golden hamsters, are not the only factors involved.


Efficacy and feasibility of the Integrated Psychological Therapy for outpatients with schizophrenia in Greece: Final results of a RCT.
The goal of this study was to evaluate the efficacy and the feasibility of cognitive remediation group therapy in patients with schizophrenia in Greece. For this purpose, the cognitive part of the Integrated Psychological Therapy (IPT), focusing on neuro- and social cognition, was compared in a randomized controlled trial (RCT) with treatment as usual (TAU). 48 outpatients took part in the study. IPT groups received 20 biweekly 1-h-therapy sessions. A test-battery was assessed at baseline, after therapy, and at a 3-month follow-up. Regarding cognitive functioning, significant effects favouring IPT were found in working memory and social perception during therapy and at follow-up. No effects could be found in verbal memory and vigilance. Significant effects favoring IPT were found in negative symptoms, in insight and in general symptoms during therapy and at follow-up using the Positive and Negative Syndrome Scale (PANSS). No effects were evident in positive symptoms and in psychosocial functioning. Significant effects favoring TAU were found in the quality of life assessment at follow-up. The study supports evidence for the feasibility and efficacy of IPT in psychiatric care in Greece and it hopefully will initiate the broader use of evidenced-based treatments like IPT in Greek Psychiatry.


Precise genome editing across kingdoms of life using retron-derived DNA.
Exogenous DNA can be a template to precisely edit a cell's genome. However, the delivery of in vitro-produced DNA to target cells can be inefficient, and low abundance of template DNA may underlie the low rate of precise editing. One potential tool to produce template DNA inside cells is a retron, a bacterial retroelement involved in phage defense. However, little effort has been directed at optimizing retrons to produce designed sequences. Here, we identify modifications to the retron non-coding RNA (ncRNA) that result in more abundant reverse-transcribed DNA (RT-DNA). By testing architectures of the retron operon that enable efficient reverse transcription, we find that gains in DNA production are portable from prokaryotic to eukaryotic cells and result in more efficient genome editing. Finally, we show that retron RT-DNA can be used to precisely edit cultured human cells. These experiments provide a general framework to produce DNA using retrons for genome modification.


Effectiveness of exercise in reducing cerebral stroke risk factors: A systematic review and meta-analysis.
This study aimed to explore the relationship between exercise and cerebral stroke and provide evidence for the prevention of cerebral stroke. All clinical trials of exercise intervention for atherosclerosis were systematically reviewed. Five major databases were searched to retrieve relevant studies from their inception to May 2022. According to the magnitude of heterogeneity, the random and fixed-effect models were used to test reasonably. According to the inclusion and exclusion criteria, 1341 articles were screened and 13 articles involving 825 patients were identified. The result showed that in the randomized controlled trials carotid intima-media thickness index was lower in the exercise group (-0.04 mm, 95% confidence interval: -0.07 to -0.01). All were statistically significant (P < .005) and subgroup analysis showed that the intervention period and paper quality are sources of heterogeneity. The results of this systematic review suggest that exercise is associated with a slow increase in carotid intima-media thickness, which may provide evidence that exercise helps reduce cerebral stroke.


Melatonin levels and microRNA (miRNA) relative expression profile in the follicular ambient microenvironment in patients undergoing in vitro fertilization process.
Intrafollicular fluid (IFF) melatonin plays a decisive role in maintaining granulosa cells' DNA integrity and protects them against apoptosis. It reduces oxidative stress and improves the oocyte quality with a higher fertilization rate. This prospective study investigated the antioxidant property of IFF melatonin and its impact on IVF outcome parameters. We also explored the relative expression of five microRNAs (miR-663b, miR-320a, miR-766-3p, miR-132-3p, miR-16-5p) and levels of cell-free DNA (cfDNA) by real-time PCR in unexplained infertile patients. We collected 425 follicular fluid (FF) samples containing mature oocytes from 295 patients undergoing IVF. Patients were subgrouped based on IFF melatonin concentration (group A ≤ 30 pg/mL, group B > 70 to ≤ 110 pg/mL, group C > 111 to ≤ 385 pg/mL). Our results showed that patients with ≤ 30 pg/mL IFF melatonin levels have significantly higher oxidative stress markers, cfDNA levels, and lower relative expression of miR-663b, miR-320a, miR-766-3p, miR-132-3p, and miR-16-5p compared to other subgroups (p < 0.001). Similarly, they have a low fertilization rate and a reduced number of high-quality day 3 embryos. Findings suggest that the therapeutic use of melatonin produces a considerable rise in the number of mature oocytes retrieved, fertilization rate, and good-quality embryo selection. Furthermore, miRNA signature enhances the quality of embryo selection, thus, may allow us to classify them as non-invasive biomarkers to identify good-quality embryos.


Efficacy of exogenous fibronectin in wound healing in malnourished rats.
Fibronectin (FN) plays an important role in fibrin matrix formation during the wound healing process. The authors investigated whether exogenous FN increases the bursting strength (BS) of surgical wounds in malnourished rats. Ninety rats were grouped according to three nutritional conditions (n = 30 in each group). All animals underwent a transverse celiotomy after 3 weeks of feeding, and FN (2 mg/body/day) was given postoperatively to 15 rats in each of the three groups. Wound BS was measured on postoperative days (PODs) 3, 5, and 7 after removal of the sutures. BS in normally nourished rats (group N; 142.6+/-23.4 mmHg) was significantly higher than that in protein malnourished rats (group PM, 110.2+/-11.2 mm Hg) and protein/ calorie (Cal)-malnourished rats (group PCM, 76.5+/-10.7 mm Hg) on POD 7 (P<.01). However, BS values for groups PM + FN (147.0+/-21.1 mmHg) and PCM + FN (115.1+/-28.9 mm Hg) were intensified significantly in comparison with groups PM and PCM (P<.01). Plasma FN levels in rats of the three FN nontreated groups were similarly decreased on POD 3 or 5, but returned to the preoperative level on POD 7, whereas those for the other three FN-treated groups increased after POD 3. Intravenous administration of FN might strengthen the weakened wounds of malnourished animals.


Updating stored memory requires adult hippocampal neurogenesis.
Adult hippocampal neurogenesis appears to influence hippocampal functions, such as memory formation for example. While adult hippocampal neurogenesis is known to be involved in hippocampal-dependent learning and consolidation processes, the role of such immature neurons in memory reconsolidation, a process involved in the modification of stored memories, remains unclear. Here, using a novel fast X-ray ablation protocol to deplete neurogenic cells, we have found that adult hippocampal neurogenesis is required to update object recognition stored memory more than to reinforce it. Indeed, we show that immature neurons were selectively recruited to hippocampal circuits during the updating of stored information. Thus, our data demonstrate a new role for neurogenesis in cognitive processes, adult hippocampal neurogenesis being required for the updating of stored OR memories. These findings suggest that manipulating adult neurogenesis may have a therapeutic application in conditions associated with traumatic stored memory, for example.


The effect of acid treatment on the structure and surface acidity of tetraalkylammonium-montmorillonites.
The effect of tetrabutylammonium (Bu4N+) and tetrapentylammonium (Pe4N+) cations on the modification of the organo-montmorillonite structure upon acid-treatment was investigated. Samples were treated with HCl for various times (2-12 h). Structural changes were followed by MAS NMR spectroscopy. The 29Si MAS NMR spectra of initial Na-saturated form (Na-SAz) showed gradual decrease of the intensity of the resonance assigned to SiO4 cross-linked in the tetrahedral sheets Q3(0Al) while signals arising from the reaction product Q31OH and Q4(0Al) became more pronounced upon acid treatment. The Q3(0Al) signal almost completely disappeared for Na-SAz treated for 8 h on contrary to Bu4N-SAz and Pe4N-SAz showing signal of relatively high intensity even after 12 h. The 27Al MAS NMR measurement proved that more than one half of Al remained in the reaction product of Bu4N-SAz and Pe4N-SAz after 8 h treatment, while Al content dropped below 5% for Na-SAz. Formation of acid sites was investigated via pyridine adsorption. Only physically adsorbed and H-bonded pyridine was detected for acid-untreated samples. In contrast, the IR spectra of the samples partially decomposed in HCl revealed bands of pyridine adsorbed on Brønsted acid sites. Strongly bonded pyridine was able to bear up heating even at 230°C.


Considerations when planning and conducting large global observational studies: Experience from the DISCOVER study of patients with type 2 diabetes.
DISCOVER is a global programme of observational research that includes patients with type 2 diabetes initiating second-line glucose-lowering therapy from 38 countries worldwide, including many with little or no previous epidemiological data available. More than 15,000 patients were followed-up for 3 years, and comprehensive data were collected using a standardized electronic case report form at enrolment, and 6, 12, 24 and 36 months. The study has formed the basis for a long-term registry that is intended to expand the geographic and clinical scope of the study and allow data collection beyond 3 years. In this review, critical aspects of study planning and implementation are summarized, along with challenges that were faced, to provide a resource for researchers planning similar studies. In particular, it is essential to set realistic expectations regarding the degree of study representativeness that can be achieved, allow for sufficient time to obtain ethics committee approval, develop tools to help recruit patients effectively, ensure that data collection systems are robust, user-friendly and adaptable, plan adequate remote and on-site monitoring, maximize patient retention through continuous engagement with study sites and ensure that everyone involved in the study forms a friendly and effective team. Observational studies such as DISCOVER are crucial for understanding disease epidemiology and management in real-world settings. They are also increasingly used by governmental, regulatory and payor agencies for post-marketing surveillance and when considering new drug submissions. The development of future studies of similar scope and ambition to DISCOVER is encouraged.


Rapid determination of SARS-CoV-2 nucleocapsid proteins based on 2D/2D MXene/P-BiOCl/Ru(bpy)32+ heterojunction composites to enhance electrochemiluminescence performance.
At the end of 2019, the novel coronavirus disease 2019 (COVID-19), a cluster of atypical pneumonia caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been known as a highly contagious disease. Herein, we report the MXene/P-BiOCl/Ru(bpy)32+ heterojunction composite to construct an electrochemiluminescence (ECL) immunosensor for SARS-CoV-2 nucleocapsid protein (CoVNP) determination. Two-dimensional (2D) material ultrathin phosphorus-doped bismuth oxychloride (P-BiOCl) is exploited and first applied in ECL. 2D architectures MXene not only act as "soft substrate" to improve the properties of P-BiOCl, but also synergistically work with P-BiOCl. Owing to the inimitable set of bulk and interfacial properties, intrinsic high electrochemical conductivity, hydrophilicity and good biocompatible of 2D/2D MXene/P-BiOCl/Ru(bpy)32+, this as-exploited heterojunction composite is an efficient signal amplifier and co-reaction accelerator in the presence of tri-n-propylamine (TPA) as a coreactant. The proposed MXene/P-BiOCl/Ru(bpy)32+-TPA system exhibits a high and stable ECL signal and achieves ECL emission quenching for "signal on-off" recognition of CoVNP. Fascinatingly, the constructed ECL biosensor towards CoVNP allows a wide linear concentration range from 1 fg/mL to 10 ng/mL and a low limit of detection (LOD) of 0.49 fg/mL (S/N = 3). Furthermore, this presented strategy sheds light on designing a highly efficient ECL nanostructure through the combination of 2D MXene architectures with 2D semiconductor materials in the field of nanomedicine. This ECL biosensor can successfully detect CoVNP in human serum, which can promote the prosperity and development of diagnostic methods of SARS-CoV-2.


Effects of antioxidant, OPC-14117, on secondary cellular damage and behavioral deficits following cortical contusion in the rat.
In the present study, we examined the effects of OPC-14117, a superoxide radical scavenger, on the secondary cellular damage and cognitive dysfunction occurring in a rat model of cerebral contusion induced by a controlled cortical impact (CCI). Histological examinations revealed that the contusion necrosis volume reached 13.6+/-5.3 mm(3) in non-treated animals and declined to 1.9+/-0.6 mm(3) in OPC-14117-treated animals (P<0.01). The cell number of the CA3 region was 120.0+/-12.4 cells/mm in the normal controls, 73.6+/-9.9 cells/mm in the non-treated animals, and 111.2+/-10.2 cells/mm in the OPC-14117-treated animals, indicating that CCI-induced selective neuronal cell death in the CA3 region was attenuated by the OPC-14117 administration (P<0.01). The tissue osmolality, as determined with a vapor pressure osmometer, was 314.5+/-15.4 mmol/kg in the normal brain and increased to 426.0+/-20.1 mmol/kg at 12 h following CCI. The increase in tissue osmolality was significantly attenuated by OPC-14117 administration (P<0.01). The OPC-14117 administration also attenuated the CCI-induced cognitive deficits. The OPC-14117-treated animals showed a tendency to improve on the Morris water maze performance test. The impairment of the habituation of exploratory activity elicited by CCI was significantly attenuated by OPC-14117 administration (P<0.05). In conclusion, OPC-14117 may have a potential for decreasing secondary cellular damage due to traumatic brain injury since it is as efficacious as any other compound tested in this model.


The Arabidopsis TFIID factor AtTAF6 controls pollen tube growth.
Initiation of transcription mediated by RNA polymerase II requires a number of transcription factors among which TFIID is the major core promoter recognition factor. TFIID is composed of highly conserved factors which include the TATA-binding protein (TBP) and about 14 TBP-associated factors (TAFs). Recently, the complete Arabidopsis TAF family has been identified. To obtain functional information about Arabidopsis TAFs, we analyzed a T-DNA insertion mutant for AtTAF6. Segregation analysis showed that plants homozygous for the mutant allele were never found, indicating that inhibition of the AtTAF6 function is lethal. Genetic experiments also revealed that the male gametophyte was affected by the attaf6 mutation since significant reduced transmission of the mutant allele through the male gametophyte was observed. Detailed histological and morphological analysis showed that the T-DNA insertion in AtTAF6 specifically affects pollen tube growth, indicating that the transcriptional regulation of only a specific subset of genes is controlled by this basal transcription factor.


Effect of narcotic prescription limiting legislation on opioid utilization following lumbar spine surgery.
Prescription opioid abuse is a public health emergency. Opioid prescriptions for spine patients account for a large proportion of use. Some states have implemented statutory limits on prescribers, however it remains unclear whether such laws are effective. This investigation compares opioid prescription patterns for patients undergoing lumbar spine surgery before and after the passage of statewide narcotic-limiting legislation in Rhode Island. Retrospective review of prospectively-collected medical and pharmacologic data. Two patient cohorts (pre-law January 1, 2016-June 31, 2016 and post-law June 1, 2017-December 31, 2017) that included all patients undergoing selected lumbar spine surgeries (lumbar discectomy, lumbar decompression without fusion, and posterior lumbar fusion). Demographic and surgical variables were collected from the patient's medical charts, and information on controlled substances was collected from the state prescription drug monitoring program database. Variables collected included the number of pills and total morphine milligram equivalents (MMEs) of the first prescription, number of prescriptions filled within 30 days of surgery, total MMEs filled in the 30-day postoperative period, and total MMEs filled from 30 to 90 days after surgery. For comparison of continuous variables, t test or Mann-Whitney U test were used as appropriate. Chi-squared analysis was utilized for comparison of categorical variables. Independent risk factors for prolonged postoperative opioid use were evaluated using logistic regression. There were no significant differences between pre-law (n = 241) and post-law (n = 311) cohorts in terms of age, sex, preoperative opioid use, or preoperative anxiolytic use (p > .05). A greater than 50% decline was observed among all patients from the pre-law to the post-law period in terms of the number of pills (51.61 vs 23.60 pills, p < .001) and MMEs (525.56 vs 218.77 MMEs, p < .001) provided in the first postoperative opioid prescription. The mean total MMEs provided in the first 30 days decreased significantly (891.26 vs 628.63 MMEs, p < .001) despite an increase in the average number of opioid prescriptions filled (1.75 vs 2.04 prescriptions, p = .002) during this time. There was no significant difference in mean MMEs filled from 30 to 90 days. Upon subgroup analysis, there was a statistically significant decline in both the mean first prescription and total 30-day MMEs regardless of preoperative opioid status (all p < .05) or specific procedure performed (all p < .05). Preoperative opioid use was strongly associated with prolonged postoperative opioid requirements throughout the study period (OR 4.71, 95% CI 3.11-7.13, p < .001). There were no significant differences between cohorts in terms of emergency department (ED) visits or unplanned hospital readmissions at 30 and 90 days following surgery (all p > .05). The institution of mandatory statewide opioid prescription limits has resulted in a significant reduction in initial and 30-day opioid prescriptions following lumbar spine surgery. Decreased opioid utilization was observed in all patients, regardless of preoperative opioid tolerance or procedure performed. No significant change in postoperative ED visits or unplanned hospital readmissions was seen following implementation of the legislation. This investigation provides preliminary evidence that narcotic limiting legislation may be effective in decreasing opioid prescriptions after lumbar spine surgery for both opioid-naïve and opioid-tolerant patients.


Characterization of the mcrBC region of Escherichia coli K-12 wild-type and mutant strains.
We have carried out an analysis of the Escherichia coli K-12 mcrBC locus in order to (1) elucidate its genetic organization, (2) to identify the proteins encoded by this region, and (3) to characterize their involvement in the restriction of DNA containing methylated cytosine residues. In vitro expression of recombinant plasmids carrying all or portions of the mcrBC region revealed that the mcrB and mcrC genes are organized as an operon. The mcrBC operon specifies five proteins, as evident from parallel in vitro and in in vivo expression studies. Three proteins of 53, 35 and 34 kDa originate from mcrB expression, while two proteins of 37 and 16 kDa arise from mcrC expression. Products of both the mcrB and mcrC genes are required to restrict the methylated substrate DNA used in this study. We also determined the nature of mutant mcrBC loci in comparison to the E. coli K-12 wild-type mcrBC locus. A major goal of these studies was to clarify the nature of the mcrB-1 mutation, which is carried by some strains employed in previous analyses of the E. coli K-12 McrBC system. Based on our analyses the mutant strains investigated could be divided into different complementation groups. The mcrB-1 mutation is a nonsense or frameshift mutation located within mcrB. It causes premature termination of mcrB gene product synthesis and reduces the level of mcrC gene expression. This finding helps to understand an existing conflict in the literature. We also describe temperature-sensitive McrA activity in some of the strains analysed and its relationship to the previously defined differences in the tolerance levels of E. coli K-12 mcrBC mutants to cytosine methylation.


Histochemical identification of osteoclasts. Review of current methods and reappraisal of a simple procedure for routine diagnosis on undecalcified human iliac bone biopsies.
Osteoclasts are known to have a high acid phosphatase content. We have adapted the simple simultaneous mono-coupling azo-dye method of Grogg and Pearse to undecalcified bone sections. A cold embedding in a mixture of glycol and methyl methacrylate was shown to well preserve the enzyme activity. Sodium alpha-naphtyl phosphate (1 mg/ml) and fast violet B (2 mg/ml) are used in 0.1 M acetate buffer, pH 5.0. The addition of 1 mM L(+) sodium tartrate selectively inhibits the acid phosphoprotein phosphatase ("osteoblastic acid phosphatase") but not osteoclastic lysosomal acid phosphatase. Counterstaining with phosphomolybdic aniline blue WS leads to well contrasted sections, providing accurate measurements of osteoclast number.


Long-term outcome of renal nerve denervation (RDN) for resistant hypertension.
We aimed to determine the long-term outcome of renal denervation (RDN). All patients with resistant hypertension who underwent RDN between 2012 and 2018 at Siriraj Hospital were included in the study. Patients were followed up at 3, 6, and 12 months and then annually up to 9 years. Effectiveness of the RDN outcome was defined by either (1) a reduction in office systolic BP ≥ 10 mmHg, (2) a reduction in the number of antihypertensive drugs taken, or (3) both outcomes being achieved. In total, 18 RDN procedures were performed during the study period. The mean and longest follow-up periods were 52 months and 104 months, respectively. Heterogeneous BP responses after RDN for resistant hypertension were observed. Effectiveness of the RDN outcome was achieved in 88% of the patients at 1 year and in >80% of the patients during the entire follow-up at each time point up to 9 years.


Surgeon Bias in the Management of Positive Sentinel Lymph Nodes.
The standard of care for clinically node-negative (cN0) patients following positive sentinel lymph node biopsy (SLNB) was completion axillary lymph node dissection (CALND). Publication of ACOSOG Z0011 in 2010 changed this standard for patients undergoing lumpectomy. Clinicians have since expanded this practice to mastectomy patients, and ongoing prospective studies are seeking to validate this practice. Here, we evaluate patient and tumor characteristics that led surgeons to forego a second surgery for CALND in cN0 mastectomy patients with positive SLNB. A single institution, retrospective review of cN0 patients with invasive primary breast cancer and positive SLNB from 2010 to 2016 was performed. Patients with T4 disease, positive preoperative axillary biopsy, prior neoadjuvant therapy or axillary surgery were excluded. Patients with positive SLNB undergoing CALND were compared with patients for whom CALND was omitted. Statistical analysis was performed using Kruskal-Wallis tests for continuous variables and χ2 tests or Fischer exact tests for categorical variables. Of 259 patients with positive SLNB, 180 (69.4%) patients underwent mastectomy. CALND was performed at the time of mastectomy in 54 (30%) patients, at time of second operation in 22 (12.2%) patients, and not performed in 104 (57%) patients. Delayed CALND was significantly associated with younger age, larger tumors, increased number of positive sentinel nodes, invasive lobular carcinoma, extranodal extension, and lymphovascular invasion. The management of cN0 patients with positive SLNB that do not meet ACOSOG Z0011 criteria is evolving and is influenced by tumor and patient characteristics in an attempt to balance the morbidity of CALND with the low rate of local regional recurrence.


Bursting through the cell cycle.
How are cells able to maintain constant levels of mRNA when the number of genes in a cell doubles ahead of cell division?


Mammographic breast density and subsequent risk of breast cancer in postmenopausal women according to the time since the mammogram.
Few studies have shown that the association between mammographic breast density and breast cancer persists for up to 10 years after the mammogram. We investigated associations of percent density, absolute dense, and nondense areas with breast cancer risk according to the time since the mammogram. This study included 1,028 incident breast cancer cases diagnosed within the Nurses' Health Study and 1,780 matched controls. Breast density was measured from digitized film images with computerized techniques. Information on breast cancer risk factors was obtained prospectively from the biennial questionnaires before the date of cancer diagnosis for cases and their matched controls. The data were analyzed with logistic regression. Breast cancer risk increased with increasing percent density and increasing absolute dense area and decreased with increasing nondense area. In multivariate analysis, the magnitude of the association between percent density and breast cancer was similar when the time since the mammogram was <2, 2 to <5, and 5 to <10 years [density ≥50% vs.<10%: ORs, 3.12; 95% confidence interval (CI): 1.55-6.25, 5.35 (95% CI: 2.93-9.76), and 3.91 (95%CI: 2.22-6.88), respectively]. Similarly, the magnitude of association between quartiles of dense and nondense areas and breast cancer risk were similar across the time strata. We found no interactions between the time since the mammogram and breast density measures (Pinteraction > 0.05). Patterns of the associations between percent density, absolute dense, and nondense area with breast cancer risk persist for up to 10 years after the mammogram. A one-time density measure can be used for long-term breast cancer risk prediction.


Cost-of-illness of melanoma in Europe - a modelling approach.
Malignant melanoma is accounting for the vast majority of skin cancer death. The treatment and productivity loss due to morbidity or premature mortality are associated with costs for society. There are few cost-of-illness (COI) studies on malignant melanoma in European countries from societal perspective and currently there is no publication analysing the COI in all European countries. The objective of the present study was to comparatively estimate COI of malignant melanoma in the European countries based on an identical approach. Cost information was obtained from results of a systematic literature research. For countries with no available cost information, a model for imputation of cost data was developed. Country-specific costs were modelled on the national gross domestic product, health expenditures, gross national income and epidemiological data. The adjustment for purchasing power parity allowed a comparison across countries. Crude national costs of malignant melanoma ranged between € 1.1 million in Iceland and € 543.8 million in Germany and resulted in € 2.7 billion for all EU/EFTA states. Estimated crude costs per patient were lowest in Bulgaria (€ 6422) and highest in Luxembourg (€ 50 734). The share of direct costs varied from 3% to 26% across countries. After adjustment for the purchasing power parity costs per patient ranged between € 14 420 in Bulgaria and € 50 961 in Cyprus. Treatment expenses and morbidity costs were markedly lower for countries that entered the EU since 2004. By contrast, mortality costs were lower in countries with a high gross domestic product per capita. In this first estimation, malignant melanoma induces relevant COI in Europe. There was large variation in the costs per patient due to different health care systems and expenses. Beyond decreasing patient burden, early intervention and prevention of melanoma could have a relevant potential to save costs across Europe.