[Morphological manifestations of compensatory-adaptive processes in the liver during aging].
Light and electron microscopy and morphometry revealed an age-related increase in the average size of hepatocytes and their nuclei in 24- and 30-month-old rats compared to 8-month-old animals, the density of hepatocytes distribution per area unit being decreased. In 24-month-old rats the number of binuclear hepatocytes increased with a subsequent decrease in their number in 30-month-old animals, which accounted for the shift in regeneration processes during ageing to predominantly intracellular one. The number of sinusoidal cells per area unit in three age groups was statistically similar. The results of morphometry and electron microscopy suggest that the compensatory-adaptive processes during hepatocyte ageing were mediated by intracellular regeneration, which led to cellular and nuclear hypertrophy similar to that observed in cells of static population (neurons, cardiomyocytes).


Hybridization by cosonication of pigeon erythrocyte membrane with exogenous lipid vesicles.
Concentrated mixtures of lipid vesicles and pigeon erythrocyte membrane were cosonicated in order to produce functional hybrid vesicles. From the properties of the resulting material, we conclude that hybrids were very probably formed. These properties were as follows: (i) The presence of membrane increased the sonic fragmentability of lipid vesicles. Sonic fragmentability was assessed by measuring sonication-induced release of previously trapped [14C]-choline and trapping of external [3H]-choline. (ii) Space enclosed by lipid was served by the membrane-like properties of 36Cl- permeability and ATP-dependent 45Ca++ uptake activity. (iii) 36Cl-permeability was more readily and fully induced into the more easily fragmented lipid vesicles. Further sonication caused loss of the induced 36Cl--permeability. This loss was less rapid with the less easily fragmented lipid vesicles; i.e., less easily fragmented lipids protected 36Cl--permeability better. (iv) Glycine uptake activity was partially protected from sonic damage by the presence of lipid vesicles. (v) On centrifugation in bovine serum albumin density gradients, cosonicated material showed lipid properties (enclosed choline and 32Pi space and [3H]-cholesterol) and membrane properties (36Cl--permeability and ATP-dependent 45Ca2+ uptake) coinciding at a density intermediate between those reached by separately sonicated membrane and lipid vesicles. (vi) Electron micrographs showed the disappearance of pure membrane-like structures and the appearance of large amounts of new vesicles whose appearance is consistent with a hybrid structure.


Thickness of medial layers of arteries prior to and during hibernation.
The thickness of the media of the aorta and the carotid, iliac, and renal arteries was measured in hibernating and nonhibernating marmots. A positive correlation was found between body weight and the thickness of the media of carotid and iliac arteries. The relation noted apparently was not due to the mean arterial pressure prior to and during hibernation, since there was no difference in the thickness of the media between nonhibernating and hibernating females. The difference due to gender could be accounted for by the greater average body weight of males. The media of the renal arteries demonstrated no relation between their thickness and body weight.


[Contribution of radiography and gammagraphy to the monitoring of the development of psychoses].
The authors emphasize the frequency of intracranial tumors in psychiatric patients. They make a distinction between the rare cases of psychiatric patients with intracranial tumors and the even most-frequent cases of tumors that express themselves through psychiatric disorders. According to the authors, the most important detection-tests are the brain scan and, more recently, the EMI scan or tomodensitometer.


Determination of the cyclic depsipeptide FK228 in human and mouse plasma by liquid chromatography with mass-spectrometric detection.
An analytical method was developed and validated for the quantitative determination of the cyclic depsipeptide FK228 (romidepsin, formerly FR901228; NSC 630176), a histone deacetylase inhibitor, in human and mouse plasma. Calibration curves were linear in the concentration range of 2-1000 ng/mL. Sample pretreatment involved a liquid-liquid extraction of 0.1 mL aliquots of plasma with ethyl acetate. FK228 and the internal standard, harmine, were separated on a Zorbax SB C18 column (75 mm x 2.1mm, 3.5 microm), using a mobile phase composed of methanol and 0.2% formic acid. The column eluent was monitored by mass spectrometry with electrospray ionization. Accuracy and precision of four concentrations of quality control samples ranged from 101.5 to 106.4% and 0.7 to 3.5% in human plasma and 93.6 to 100.6% and 0.6 to 6.5%, in mouse plasma, respectively. This method represents a significant improvement over our previously published analytical assay for this agent, decreasing the sample volume requirements, increasing the accuracy and precision (through addition of a suitable internal standard), expanding the analytical range and validating in additional biological matrices. The developed method was applied to study the pharmacokinetics of FK228 in over 1000 clinical and preclinical samples.


Age-dependent conformational changes in acid deoxyribonuclease of chick brain.
Acid deoxyribonuclease (EC 184.108.40.206) (DNase) from young (16 days of incubation) and old (1.5 years) chick cerebral hemispheres was purified to apparent homogeneity. Throughout the purification schedule, the behavior of "young" and "old" enzymes was similar. However, the specific activity of the purified enzyme from old brain was only one-tenth that of young enzyme. Polyacrylamide gel electrophoresis of the purified acid DNase gave a single band. Antisera against both "young" and "old" enzyme were raised and double immunodiffusion experiments revealed cross-reaction of young antigen with old antiserum and vice versa, although precipitin bands with young antigen against young antiserum and old antigen against old antiserum were more sharp. Both young and old acid DNase preparations showed an apparent molecular weight of 62,000 and many other properties like heat stability, effect of various exogenous compounds like Hg2+, Zn2+, Mg2+, etc., were also similar. The old enzyme showed slightly higher Km and decreased Vmax compared with the young enzyme. Dansylation of N-terminal amino acids and their analysis following tryptic digestion of both "young" and "old" acid DNase revealed a similar pattern. Immunotitration experiments showed that the old enzyme requires more antiserum prepared against "young" enzyme to achieve 50% inactivation, thus pointing out the presence of completely or partially inactive molecules in "old" acid DNase preparation. Circular dichroism spectra of the enzyme preparations indicated that the "old" acid DNase molecules are more rigid and have more alpha-helical structure, compared with the "young" enzyme. From these data, it is suggested that the reduction in the specific activity of old acid DNase may be, apart from other possibilities, due to conformational changes in the enzyme molecules.


Association of renal adenocarcinoma and BK virus nephropathy post transplantation.
While most BK virus infections are asymptomatic, immunosuppression has been associated with BK virus reactivation and impaired graft function or ureteric ulceration in renal transplant patients and hemorrhagic cystitis in bone marrow transplant patients. Oncogenicity is also postulated and this is the first report of a child with a carcinoma of the donor renal pelvis following BK virus allograft nephropathy. Removal of the primary tumor and cessation of immunosuppression led to regression of secondary tumors and a return to health.


Studies on the expression of Cu,Zn superoxide dismutase in human tissues during development.
The developmental expression of Cu,Zn superoxide dismutase in human lung and erythrocytes has been studied using activity measurements, immunoblotting and immunohistochemistry. Enzyme activity in erythrocytes increased significantly during gestation but no developmental trend was seen in lung. Immunoblotting identified a single enzyme form that was present in a variety of tissues and immunohistochemistry showed the enzyme to have widespread distribution in lung tissue. These data indicate that Cu,Zn superoxide dismutase is consistently expressed during human development and that, unlike in other species, no late-fetal surge in expression occurs.


Phenotypic characterization of chondrosarcoma-derived cell lines.
Gene expression profiling of three chondrosarcoma derived cell lines (AD, SM, 105KC) showed an increased proliferative activity and a reduced expression of chondrocytic-typical matrix products compared to primary chondrocytes. The incapability to maintain an adequate matrix synthesis as well as a notable proliferative activity at the same time is comparable to neoplastic chondrosarcoma cells in vivo which cease largely cartilage matrix formation as soon as their proliferative activity increases. Thus, the investigated cell lines are of limited value as substitute of primary chondrocytes but might have a much higher potential to investigate the behavior of neoplastic chondrocytes, i.e. chondrosarcoma biology.


Cholesterogenesis and cell division in phytohemagglutinin-stimulated human lymphocytes: a comparative study with several inhibitors.
It has been shown that when lymphocytes are stimulated by phytohemagglutinin the expected stimulation of DNA synthesis is preceded by stimulation of cholesterol synthesis. This confirms the existence of a relation between cell division and cholesterol synthesis. We studied the effect on cell division of six inhibitors of cholesterol biosynthesis, previously shown to interfere with different steps of the process: 7 beta-hydroxycholesterol, 25-hydroxycholesterol, lanost-7-en-3 beta, 32-diol, mevinolin, propiconazole, dodecylimidazole. Since experiments were performed in the presence of a high percentage of human serum, which provided cells with exogenous cholesterol via the LDL-receptor pathway, our investigation was focused on the role of newly synthesized cholesterol. The biosynthesis was evaluated by labeling cells with [14C]sodium acetate; to take into account variations of cell permeability to sodium acetate, the results were expressed as the percentage of total cellularly incorporated radioactivity transformed into cholesterol, after separation from all other labeled metabolites. These data were compared with the percentage of transformation into nonsaponifiable lipids, which varied in parallel with HMG-CoA reductase activity, as confirmed by direct enzymatic measurement. Cell division was assessed by simultaneous measurements of three parameters: thymidine incorporation into DNA, cell proliferation and cellular protein content. All the effectors strongly inhibited the conversion of labeled acetate into cholesterol, but cell division was not inhibited by two of them: propiconazole and 7 beta-hydroxycholesterol. These compounds only slightly inhibited the synthesis of nonsaponifiable lipids, which mainly consisted of methylsterols resulting from a blockage of lanosterol demethylation. Thus, it can be concluded that the nonsaponifiable metabolite essential for cell growth is not newly synthesized cholesterol. It was also found that inhibitors affected cell division only when they were added to the culture medium before the decline of cholesterol synthesis stimulation.


Structure-activity relationship of ligands of dihydrouracil dehydrogenase from mouse liver.
One hundred and five nucleobase analogues were screened as inhibitors of dihydrouracil dehydrogenase (DHUDase, EC 126.96.36.199) from mouse liver. 5-Benzyloxybenzyluracil, 1-deazauracil (2,6-pyridinediol), 3-deazauracil (2,4-pyridinediol), 5-benzyluracil, 5-nitrobarbituric acid and 5,6-dioxyuracil (alloxan) were identified as potent inhibitors of this activity, with apparent Ki values of 0.2, 0.5, 2.1, 3.4, 3.8 and 6.6 microM respectively. Both 5-benzyloxybenzyluracil and 1-deazauracil were also potent inhibitors of DHUDase from human livers. These findings along with an extensive review of literature allowed the formulation of a structure-activity relationship. The binding to DHUDase required intact C2 and C4 oxo groups. Replacement of N1 or N3 by an endocyclic carbon enhanced binding. In contrast, replacement of C5 or C6 by an endocyclic nitrogen abolished binding. Addition of a charged group to C5 and/or C6, and of a hydrophobic group to C5 but not C6 improved the binding.


Novel approach for genome scan meta-analysis of rheumatoid arthritis: a kernel-based estimation procedure.
Genome scan meta-analysis (GSMA) can prove very useful in detecting genetic effects too small to be detected in an individual linkage study and can also lead to more consistent results. In this paper, we propose a new kernel-based estimation procedure for GSMA. Instead of estimating identity by descent between markers, as performed in interval mapping approaches, we estimated directly the nonparametric linkage score between markers using a kernel procedure. The GSMA is then extended to take into account the kernel estimate of the nonparametric linkage score and its variance at a given chromosomal position. The method is applied to the rheumatoid arthritis genome scan data (Genetic Analysis Workshop 15 Problem 2).


Effects of 10 T static magnetic field on the function of sperms and their offspring in Caenorhabditis elegans.
With the wide application of static magnetic fields (SMFs), the risk of living organisms exposed to man-made magnetic fields that the intensity is much higher than geomagnetic field has gradually increased. Reproductive system is highly sensitive to environmental stress; however, the influence of high SMFs on reproduction system is still largely unknown. Here we explored the biological responses of SMFs exposure at an intensity of 10 T on the sperms and their offspring in him-5 male mutants of Caenorhabditis elegans (C. elegans). The size of unactivated sperms was deceased by 10 T SMF exposure, instead of the morphology. Exposure to 10 T SMF significantly altered the function of sperms in him-5 worms including the activation of sperms and the non-transferred ratio of sperms. In addition, the brood size assay revealed that 10 T SMF exposure eventually diminished the reproductive capacity of him-5 male worms. The lifespan of outcrossed offspring from exposed him-5 male mutants and unexposed fog-2 female mutants was decreased by 10 T SMF in a time dependent manner. Together, our findings provide novel information regarding the adverse effects of high SMFs on the sperms of C. elegans and their offspring, which can improve our understanding of the fundamental aspects of high SMFs on biological system.


Structural insight into the selective agonist ST1936 binding of serotonin receptor 5-HT6.
The serotonin receptor 5-HT6R is an important G-protein-coupled receptor (GPCR) that involved in essential functions within the central and peripheral nervous systems and is linked to various psychiatric disorders. Selective activation of 5-HT6R promotes neural stem cell regeneration activity. As a 5-HT6R selective agonist, 2-(5 chloro-2-methyl-1H-indol-3-yl)-N, N-dimethylethanolamine (ST1936) has been widely used to investigate the functions of the 5-HT6R. The molecular mechanism of how ST1936 is recognized by 5-HT6R and how it effectively couples with Gs remain unclear. Here, we reconstituted the ST1936-5-HT6R-Gs complex in vitro and solved its cryo-electron microscopy structure at 3.1 Å resolution. Further structural analysis and mutational studies facilitated us to identify the residues of the Y3107.43 and "toggle switch" W2816.48 of the 5-HT6R contributed to the higher efficacy of ST1936 compared with 5-HT. By uncovering the structural foundation of how 5-HT6R specifically recognizes agonists and elucidating the molecular process of G protein activation, our discoveries offer valuable insights and pave the way for the development of promising 5-HT6R agonists.


The important role of solvent vapor in an organic solid state reaction.
Some organic reactions in the solid state proceeded very efficiently and selectively in the presence of a small amount of solvent vapor.


Impact of intrinsic and extrinsic risk factors on early-onset lung disease in cystic fibrosis.
Although respiratory pathology is known to develop in young children with cystic fibrosis (CF), the determinants of early-onset lung disease have not been elucidated. We aimed to determine the impact of potential intrinsic and extrinsic risk factors during the first 3 years of life, testing the hypothesis that both contribute significantly to early-onset CF lung disease. We studied 104 infants born during 2012-2017, diagnosed through newborn screening by age 3 months, and evaluated comprehensively to 36 months of age. Lung disease manifestations were quantified with a new scoring system known as CFELD for Cystic Fibrosis Early-onset Lung Disease. The variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene were determined and categorized. Whole genome sequencing was performed on each subject and the data transformed to polygenic risk scores (PRS) that aggregate variants associated with lung function. Extrinsic factors included socioeconomic status (SES) indicators and environmental experiences such as exposures to smoking, pets, and daycare. We found by univariate analysis that CFTR genotype and genetic modifiers aggregated by the PRS method were significantly associated with early-onset CF lung disease. Ordinal logistic regression analysis demonstrated that high and stable SES (maternal education ≥community college, stable 2-parent home, and not receiving Medicaid) and better growth (weight-for-age and height-for-age z-scores) reduced risks, while exposure to smoking and daycare ≥20 h/week increased the risk of CFELD severity. Extrinsic, modifiable determinants are influential early and potentially as important as the intrinsic risk factors in the onset of CF lung disease.


The Architecture of the Human RNA-Binding Protein Regulatory Network.
RNA-binding proteins (RBPs) are key players of post-transcriptional regulation of gene expression, relying on competitive and cooperative interactions to fine-tune their action. Several studies have described individual interactions of RBPs with RBP mRNAs. Here we present a systematic network investigation of fifty thousand interactions between RBPs and the UTRs of RBP mRNAs. We identified two structural features in this network. RBP clusters are groups of densely interconnected RBPs co-binding their targets, suggesting a tight control of cooperative and competitive behaviors. RBP chains are hierarchical structures connecting RBP clusters and driven by evolutionarily ancient RBPs. These features suggest that RBP chains may coordinate the different cell programs controlled by RBP clusters. Under this model, the regulatory signal flows through chains from one cluster to another, implementing elaborate regulatory plans. This work thus suggests RBP-RBP interactions as a backbone driving post-transcriptional regulation of gene expression to control RBPs action on their targets.


Effectiveness of a Suicide Prevention Module for Adults in Substance Use Disorder Treatment: A Stepped-Wedge Cluster-Randomized Clinical Trial.
Individuals with substance use disorders (SUDs) are at high risk for suicide. The Preventing Addiction Related Suicide (PARS) module is the first suicide prevention module developed in and for community substance use intensive outpatient programs (IOPs). To evaluate the effectiveness of PARS on suicide-related outcomes (ie, knowledge, attitudes, and help-seeking behavior) compared with usual care. This stepped-wedge cluster-randomized clinical trial was conducted from 2017 to 2020, with follow-up assessments conducted after treatment and at 1, 3, and 6 months. Participants included adult outpatients in SUD treatment at community IOPs across western Washington state. Data were analyzed from July 1, 2020, to January 20, 2022. The intervention, PARS, was a 1-session secondary prevention module administered by trained SUD counselors consisting of didactic presentations and group discussions about suicide risk factors, warning signs, and actions to take if suicide risk is observed in self or others. The control group received usual care. Primary outcomes were suicide knowledge, attitudes about suicide, and help-seeking behavior among patients enrolled in an IOP. A total of 906 participants (mean [SD] age, 37.5 [12.0] years; 540 [59.6%] men) were included, with 478 participants receiving usual care and 428 participants receiving PARS. In intent-to-treat analysis from baseline to after treatment, there was a greater improvement in suicide knowledge (d = 0.15; 95% CI, 0.08 to 0.23; P < .001) and a greater reduction in maladaptive attitudes (d = 0.18; 95% CI, 0.14 to 0.25; P < .001) for PARS participants compared with those receiving usual care. Improvements were maintained at follow-up for suicide knowledge (1 month: d = 0.16; 95% CI, 0.07 to 0.22; P < .001; 3 months: d = 0.12; 95% CI, 0.05 to 0.19; P = .001; 6 months: d = 0.13; 95% CI, 0.06 to 0.20; P < .001) and reductions in maladaptive attitudes (1 month: d = 0.20; 95% CI, 0.12 to 0.23; P < .001; 3 months: d = 0.10; 95% CI, 0.05 to 0.16; P < .001; 6 months: d = 0.14; 95% CI, 0.09 to 0.19; P < .001), with 788 participants (87.0%) of the sample responding across time points. From baseline to 6 months, there was a greater improvement in help-seeking in the PARS group vs usual care (d = 0.16; 95% CI, 0.01 to 0.32; P = .04). This stepped-wedge cluster-randomized clinical trial found that PARS was superior to usual care in improving suicide knowledge, maladaptive attitudes, and help-seeking in adults undergoing community addiction treatment. As a 1-session IOP module developed in partnership with community addiction agencies, PARS has the potential for wide impact in the national suicide prevention strategy. ClinicalTrials.gov Identifier: NCT03166709.


Piezoelectric quartz crystal sensor for rapid analysis of pirimicarb residues using molecularly imprinted polymers as recognition elements.
A new piezoelectric quartz crystal (PQC) sensor using molecularly imprinted polymer (MIP) as sensing material has been developed for fast and onsite determination of pirimicarb in contaminated vegetables. Three MIPs particles have been prepared by conventional bulk polymerization (MIP-B) and precipitation polymerization in either acetonitrile (MIP-P1) or chloroform (MIP-P2). MIP-P2, with uniform spherical shape and mean diameter at about 50 nm, has shown the best performance as the sensing material for PQC sensor. The sensor fabricated with MIP-P2 can achieve a steady-state response within 5 min, a very short response time as compared to MIPs-coated PQC sensor reported in the literature. The sensor developed exhibits good selectivity (low response to those pesticides with similar structures to pirimicarb, such as atrazine, carbaryl, carbofuran and aldicarb) and high sensitivity to pirimicarb with a linear working range from 5.0 x 10(-6) to 4.7 x 10(-3) mol L(-1) (following a regression equation (r=0.9988) of -DeltaF=0.552+1.79 x 10(6) C), a repeatability (R.S.D., n=5) of 4.3% and a detection limit (S/N=3, n=5) of 5 x 10(-7) mol L(-1). The MIP-coated PQC sensor developed is shown to provide a sensitive and fast method for onsite determination of pirimicarb in aqueous extract from contaminated vegetables with satisfactory recoveries from 96 to 103% and repeatability (R.S.D., n=5) from 4.6 to 7.1% at pirimicarb concentrations ranging from 8.0x10(-6) to 2.0 x 10(-4) mol L(-1).


Deleterious brain cell membrane effects after NMDA receptor antagonist administration to newborn piglets.
Previous studies have shown that administration of the N-methyl-d-aspartate (NMDA) receptor antagonist 3-(2-carboxypiperazin-4-yl)-1-phosphonic acid (CPP) reduces NMDA-mediated neurotoxicity in animal models of hypoxia/ischemia but also may induce brain tissue vacuolization and alter glucose metabolism. The present study tests the hypothesis that CPP administration alters brain cell membrane structure and function in the cerebral cortex of normoxic newborn piglets through the generation of oxygen free radicals and induction of lipid peroxidation. Twenty six anesthetized, ventilated newborn piglets-13 treated with 2 mg/kg i.v. CPP and 13 untreated controls-were studied. ATP and phosphocreatine (PCr) levels were measured as an index of cellular energy metabolism and tissue glucose levels determined. Na+, K+-ATPase activity was measured as an index of brain cell membrane function and the lipid peroxidation products conjugated dienes (CD) and fluorescent compounds (FC) measured. Free radical generation was detected on cortical biopsies homogenized with alpha-phenyl-N-tert-butyl-nitrone (PBN) through electron spin resonance spectroscopy. Signal height of spectrum was divided by dry tissue weight and expressed as mm/g tissue. In the two groups brain tissue ATP and PCr levels were not different. Tissue glucose levels were higher in the CPP group (24+/-5 mg/dl) than in controls (14+/-3 mg/dl), p<0.05, whereas Na+,K+-ATPase activity was lower in the CPP group than in controls (34+/-4 vs. 43+/-6 micromol Pi/mg protein/h), p<0.05. Lipid peroxidation products were higher in the CPP group (CD: 57+/-19 nmol/g brain, FC: 1.5+/-0.3 microg/g brain) than in controls (CD: 0+/-0 nmol/g brain, FC: 0.9+/-0.2 microg/g brain), p<0. 05. Free radical intensity was higher in the CPP group (493+/-397 mm/g tissue) than in controls (51+/-83 mm/g tissue), p<0.05. In vitro administration of CPP to brain cell membranes did not change Na+,K+-ATPase activity or the generation of lipid peroxidation products. The data demonstrate that administration of CPP induces lipid peroxidation, results in free radical generation, decreases brain cell membrane Na+,K+-ATPase activity and alters glucose metabolism in the cerebral cortex of newborn piglets. Since CPP is a potent antagonist of the NMDA receptor, we speculate that CPP generates free radicals through a pathway independent of the NMDA receptor by altering cellular metabolism and possibly glucose utilization during normoxia in newborn piglets.


"I Suck at Everything": Crime, Arrest, and the Generality of Failure.
In this paper we advance the idea that getting arrested amounts to "failing at crime." And akin to the notion of the generality of deviance-where those who engage in any given form of criminal behavior are also likely to engage in a wide array of other problematic behaviors-we examine whether failing at crime (getting arrested) is associated with other forms of life failure. Using data from multiple waves of the National Longitudinal Study of Adolescent to Adult Health, our results reveal that, independent of one's level of self-reported criminal behavior and other key potential confounders (IQ and self-control), being arrested is a significant predictor of a host of life failures related to education, employment, relationships, and health. The key implication of our study is that it highlights the need to develop a theory of the "generality of failure."


In vivo release of cholecystokinin-like immunoreactivity in rat frontal cortex under freely moving conditions.
Release of cholecystokinin-like immunoreactivity (CCK-LI) was measured in rat medial prefrontal cortex in vivo by brain dialysis and enzyme immunoassay under freely moving condition. Perfusion of 50 mM K+ resulted in the increase of CCK-LI in the dialysate. The data on high-performance liquid chromatography (HPLC) of the dialysate showed that the increase of the CCK-LI was mainly due to the increase of CCK octapeptide sulfate itself. In the preliminary experiments, we applied this brain dialysis method for determination of CCK-LI release with drug treatment. After treatment with sulpiride (i.p.), a D2 dopamine receptor antagonist, a significant increase of CCK-LI was observed, indicating that this brain dialysis technique is applicable to detect change in the level of CCK-LI release after a certain drug treatment.


Filling the Organ Donor Pool by Giving Priority.
Many of the alternatives for procuring donor organs are considered either ineffective, unethical, or illegal. One possibility that may not face such challenges is a priority system whereby individuals who register as an organ donor are given priority to receive an organ over those who have not registered. However, providing extrinsic rewards can sometimes paradoxically reduce the target behavior, especially for those who are more altruistically motivated. Two behavioral experiments were employed and data were analyzed using regressions as well as examining open-ended responses. The results suggest that giving priority to receive an organ to those who register to donate postmortem could increase overall registration rates. Further, the effect of providing priority appears to work by inducing anticipated regret, which can be used to overcome common obstacles to registration. Finally, it was found that a priority system is most effective in increasing donor rates for those individuals who are less altruistically motivated and does not reduce registration rates for those who are more altruistically motivated. Given the unabated shortage of transplant organs, the finding that a priority system could increase the willingness to register as a donor without crowding out altruistically motivated individuals is highly encouraging.


Predictive value of intrathecal interleukin-6 for ventriculostomy-related Infection.
Early diagnosis of ventriculostomy-related infection (VRI) is crucial for the early treatment and course of this disease. In neurosurgical patients the diagnostic criteria are equivocal, mostly because of bloodstained cerebrospinal fluid (CSF). The predictive value, sensitivity and specificity of intrathecal interleukin-6 (IL-6 (CSF)) has been proven for VRI compared with classical diagnostic CSF parameters, i.e. cell countCSF (CC (CSF)) and total protein (CSF). We prospectively analyzed the daily clinical data and CSF samples of 75 neurosurgical patients with an external ventricular drainage (EVD), which had been inserted predominantly because of poor-grade subarachnoid hemorrhage (SAH). The intrathecal interleukin-6 concentrations (IL-6 (CSF)) were correlated with the clinical course and VRI incidence, as diagnosed by the classical VRI criteria (CC (CSF), total protein (CSF), clinical symptoms). Based on classical criteria, bacterial meningitis occurred in 26.7% of patients. Patients with VRI manifested significantly (p<0.001) higher median values of IL-6 (CSF) (up to 2,000-fold increase) the day before (day -1) infection was diagnosed by conventional parameters. Using a cut-off value of IL-6 (CSF)>or=2,700 pg/ml [4,050 pg/ml after WHO standardization] on day -1, the relative risk for VRI was 6.09 (95% CI: 2.62-14.18%). A predictive value of IL-6 (CSF)>or=2,700 pg/ml [4,050 pg/ml] for VRI was calculated of 89% (95% CI: 79.6-98.0%), a sensitivity of 73.7% and a specificity of 91.4%. The amount of intrathecal blood was an independent risk factor for VRI occurrence, whereas the mean duration of EVD in place showed no impact on the rate of infection. Our data indicate that IL-6 (CSF) is a reliable marker for predicting VRI prior to clinically manifest meningitis, one day earlier than the common diagnostic criteria of CSF infection (CC (CSF), total protein (CSF), clinical symptoms).


Clinical characteristics and outcomes in patients with metastatic breast cancer and pseudocirrhosis: a single center retrospective cohort study.
Pseudocirrhosis is a term used to describe changes in hepatic contour that mimic cirrhosis radiographically, but lack the classic pathologic features of cirrhosis. This radiographic finding is frequently found in patients with metastatic breast cancer (MBC), but the risk factors and clinical consequences are poorly understood. In this retrospective study, we identified patients with MBC and pseudocirrhosis who were treated at a single center from 2002 to 2021. We used chart extraction and radiology review to determine demographic characteristics, treatment history, imaging features, and complications of pseudocirrhosis. 120 patients with MBC and pseudocirrhosis were identified with the following BC subtypes: hormone receptor (HR) positive, HER2 negative (n = 99, 82.5%), HR+/HER2+ (n = 14, 11.7%), HR- /HER2+ (n = 3, 2.5%), and triple negative (TNBC; n = 4, 3.3%). All patients had liver metastases and 82.5% (n = 99) had > 15 liver lesions. Thirty-six patients (30%) presented with de novo metastatic disease. Median time from MBC diagnosis to pseudocirrhosis was 29.2 months. 50% of patients had stable or responding disease at the time of pseudocirrhosis diagnosis. Sequelae of pseudocirrhosis included radiographic ascites (n = 97, 80.8%), gastric/esophageal varices (n = 68, 56.7%), splenomegaly (n = 26, 21.7%), GI bleeding (n = 12, 10.0%), and hepatic encephalopathy (n = 11, 9.2%). Median survival was 7.9 months after pseudocirrhosis diagnosis. Radiographic ascites was associated with shorter survival compared to no radiographic ascites (42.8 vs. 76.2 months, p = < 0.001). This is the largest case series of patients with MBC and pseudocirrhosis. Nearly all patients had HR+ MBC and extensive liver metastases. Survival was short after pseudocirrhosis and prognosis worse with radiographic ascites.


Diet-induced changes in sex hormone binding globulin and free testosterone in women with normal or polycystic ovaries: correlation with serum insulin and insulin-like growth factor-I.
The purpose of this study was to investigate the effect of calorie restriction on serum concentrations of sex hormone binding globulin (SHBG) in women with normal or polycystic ovaries (PCO) and to examine the possible role of insulin and insulin-like growth factor-I (IGF-I) in mediating changes in SHBG levels. Six normal subjects with mean (SD) body mass index (BMI) 25.5 (2.2) and five subjects with PCO (BMI 36.1 (3.7)) were studied before and after 2 or (PCO only) 4 weeks of a very low calorie diet (330 kcal/day; Cambridge Diet). In both normal women and patients with PCO there was a twofold increase in SHBG concentrations after 2 weeks and this was sustained in the PCO subjects for a further 2 weeks. The rise in SHBG was accompanied by a fall in free testosterone concentrations. There were parallel changes in serum insulin and IGF-I concentrations which decreased during the diet and there were significant negative correlations of SHBG with insulin in both normal subjects (r = -0.62) and women with PCO (r = -0.60). In addition, serum concentrations of an insulin-dependent small molecular weight (34 kDa) binding protein for IGF-I (IGF-BPI) increased significantly during dieting in both groups and were negatively correlated with serum insulin (controls, r = -0.56; PCO, r = -0.68) and positively correlated with serum SHBG levels (controls, r = 0.69; PCO, r = 0.63). In summary, these data indicate that in both normal subjects and those subjects with PCO, calorie restriction results in a highly significant increase in SHBG concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)


Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity.
Development of a highly reproducible and sensitive single-cell RNA sequencing (RNA-seq) method would facilitate the understanding of the biological roles and underlying mechanisms of non-genetic cellular heterogeneity. In this study, we report a novel single-cell RNA-seq method called Quartz-Seq that has a simpler protocol and higher reproducibility and sensitivity than existing methods. We show that single-cell Quartz-Seq can quantitatively detect various kinds of non-genetic cellular heterogeneity, and can detect different cell types and different cell-cycle phases of a single cell type. Moreover, this method can comprehensively reveal gene-expression heterogeneity between single cells of the same cell type in the same cell-cycle phase.


Radiological approach to basilar invagination type B: Reliability and accuracy.
This study aims to determine the reliability of the radiological tests used in the diagnosis of basilar invagination (BI). Patients diagnosed with type B basilar invagination, who had both magnetic resonance (MR) and computed tomography (CT) imaging between January 2014 and November 2019 were included in this retrospective reliability study. In this study, distance from odontoid apex to Chamberlain's line (OA-CL) was accepted as a reference method for the diagnosis. Forty-two BI cases and 79 controls were included. Three radiologists with different levels of experience individually evaluated OA-CL, Boogard's angle (BoA), clivo-axial angle (CXA), clivo-dens angle (CDA), and clivo-palate angle (CPA) on midsagittal CT and MR images. Statistical analysis was made with the intraclass correlation coefficient (ICC), t-test, and receiver operating characteristic (ROC) curve. The ICC for CT and MR were; 0.977-0.973 (OA-CL), 0.912-0.882 (BoA), 0.845-0.846 (CXA), 0.862-0.864 (CDA), and 0.762-0.747 (CPA) respectively (P < 0.001). The areas under the ROC curve were 0.977 (BoA), 0.832 (CXA), 0.852 (CDA), and 0.719 (CPA) (P < 0.001). The cut-off measures were ≥137.84° (BoA), ≤149.25° (CXA), ≤129.58° (CDA), and ≤61.83° (CPA). The diagnostic accuracies were 0.954 (BoA), 0.664 (CXA), 0.704 (CDA), 0.438 (CPA) (P < 0.001). OA-CL and BoA express excellent inter-rater agreement than CXA, CDA, and CPA, which are limited due to morphological variations and head spatial position. BoA is the second most reliable diagnostic test. CXA, CDA, should only be used for complementary information. CPA was found inadequate for the diagnosis of BI..


Aqueous two-phase systems: A simple methodology to obtain mixtures enriched in main toxins of Bothrops alternatus venom.
Phospholipase A2 (PLA2) and protease (P) are enzymes responsible of myotoxic, edematogenic and hemostasis disorder effects observed in the envenomation by Bothrops alternatus pitviper. Their partitioning coefficient (Kp) in different polyethyleneglycol/potassium phosphate aqueous two-phase systems (ATPSs) was determined in order to both achieve a better understanding of the partitioning mechanism and define optimal conditions for toxin isolation. Polyethyleneglycols (PEGs) of molecular weights 1000; 3350; 6000 and 8000; different temperatures (5, 20 and 37 °C) and phase volume ratios of 0.5; 1 and 2 were assayed. PLA2 partitioned preferentially to the top phase while P mainly distributed to the bottom phase. Either entropically- or enthalpically-driven mechanisms were involved in each case (PLA2 and P). The aqueous two-phase system formed by PEG of MW 3350 (12.20% wt/wt) and KPi pH 7.0 (11.82% wt/wt) with a volume ratio of one and a load of 1.25 mg of venom/g of system showed to be the most efficient to recover both enzymes. It allowed obtaining the 72% of PLA2 in the top phase with a purification factor of 2 and the 82% of P at the bottom phase simultaneously. A further adsorption batch step with DEAE-cellulose was used to remove satisfactorily the PEG from the top phase and recover the active PLA2. The proposed methodology is simple, inexpensive, and only requires professionals trained in handling basic laboratory equipment. It could be easily adoptable by developing countries in which the snakebite accidents cause considerable morbidity and mortality.


Periocardiophrenic Release to Prevent Conduit Compression After Neonatal Truncus Arteriosus Repair.
Sternal approximation postcardiac surgery in neonates can sometimes be challenging. Neonatal truncus arteriosus repair using a right ventricle-to-pulmonary artery homograft conduit is one such surgical procedure wherein there is a risk of developing conduit compression after sternal closure. We describe our technique of pericardiophrenic release at the time of delayed sternal closure to prevent hemodynamic compromise and conduit compression after sternal approximation in such cases.


Preparation of thermo-responsive graft copolymer by using a novel macro-RAFT agent and its application for drug delivery.
A methodology to prepare thermo-responsive graft copolymer by using a novel macro-RAFT agent was proposed. The macro-RAFT agent with pendant dithioester (ZC(S)SR) was facilely prepared via the combination of RAFT polymerization and esterification reaction. By means of ZC(S)SR-initiated RAFT polymerization, the thermo-responsive graft copolymer consisting of poly(methyl methacrylate-co-hydroxylethyl methacrylate) (P(MMA-co-HEMA)) backbone and hydrophilic poly(N-isopropylacrylamide) (PNIPAAm) side chains was constructed through the "grafting from" approach. The chemical compositions and molecular weight distributions of the synthesized polymers were respectively characterized by (1)H nuclear magnetic resonance ((1)H NMR) and gel permeation chromatography (GPC). Self-assembly behavior of the amphiphilic graft copolymers (P(MMA-co-HEMA)-g-PNIPAAm) was studied by transmission electron microscopy (TEM), dynamic light scattering (DLS) and spectrofluorimeter. The critical micelle concentration (CMC) value was 0.052 mg mL(-1). These micelles have thermo-responsibility and a low critical solution temperature (LCST) of 33.5°C. Further investigation indicated that the guest molecule release property of these micelles, which can be well described by a first-order kinetic model, was significantly affected by temperature. Besides, the micelles exhibited excellent biocompatibility and cellular uptake property. Hence, these micelles are considered to have potential application in controlled drug delivery.


Community-based strategies for immunizing the "hard-to-reach" child: the New York State immunization and primary health care initiative.
The 1989-1991 measles epidemic in New York City drew attention to the low immunization coverage rates found in urban neighborhoods. This article describes a joint initiative of the New York State Department of Health and the Columbia University School of Public Health to mobilize parents to fully immunize their children. Eleven community-based organizations (CBOs) used a variety of outreach strategies to identify and enroll underimmunized children in primary care. They enrolled 4,555 children, of whom 75% needed at least one basic vaccine dose to be up-to-date for their age. Enrolled children were followed by CBOs to ensure compliance with appointments. After nine months of program operation, 73% of children in an evaluation sample were up-to-date for age for their immunizations. Immunization coverage increases were greatest for the youngest children, for whom coverage rates more than doubled in the first nine months of program operation. Ninety-one percent of these "hard to reach" children were tracked successfully by CBOs. This article compares the strategies used by the community organizations and concludes with suggestions for improvements of future community-based mobilization programs.


Motivation and incentives of rural maternal and neonatal health care providers: a comparison of qualitative findings from Burkina Faso, Ghana and Tanzania.
In Burkina Faso, Ghana and Tanzania strong efforts are being made to improve the quality of maternal and neonatal health (MNH) care. However, progress is impeded by challenges, especially in the area of human resources. All three countries are striving not only to scale up the number of available health staff, but also to improve performance by raising skill levels and enhancing provider motivation. In-depth interviews were used to explore MNH provider views about motivation and incentives at primary care level in rural Burkina Faso, Ghana and Tanzania. Interviews were held with 25 MNH providers, 8 facility and district managers, and 2 policy-makers in each country. Across the three countries some differences were found in the reasons why people became health workers. Commitment to remaining a health worker was generally high. The readiness to remain at a rural facility was far less, although in all settings there were some providers that were willing to stay. In Burkina Faso it appeared to be particularly difficult to recruit female MNH providers to rural areas. There were indications that MNH providers in all the settings sometimes failed to treat their patients well. This was shown to be interlinked with differences in how the term 'motivation' was understood, and in the views held about remuneration and the status of rural health work. Job satisfaction was shown to be quite high, and was particularly linked to community appreciation. With some important exceptions, there was a strong level of agreement regarding the financial and non-financial incentives that were suggested by these providers, but there were clear country preferences as to whether incentives should be for individuals or teams. Understandings of the terms and concepts pertaining to motivation differed between the three countries. The findings from Burkina Faso underline the importance of gender-sensitive health workforce planning. The training that all levels of MNH providers receive in professional ethics, and the way this is reinforced in practice require closer attention. The differences in the findings across the three settings underscore the importance of in-depth country-level research to tailor the development of incentives schemes.


Classic illustrations of benign and malignant phyllodes breast tumors in two patients.
Phyllodes tumors, World Health Organization fibroepithelial tumors, are classified as benign, borderline, or malignant based on histopathology. Phyllodes must be distinguished from benign fibroadenomas, also WHO fibroepithelial tumors. The distinction of phyllodes from fibroadenomas can be challenging clinically, as these tumors may mirror one another. Here, we present 2 cases, classic clinical and imaging examples of benign and malignant phyllodes, to review the current epidemiology, classification, diagnosis, and treatment of phyllodes tumors.


Highly diastereoselective formation of spirocyclic compounds via 1,5-hydrogen transfer: a total synthesis of (-)-erythrodiene.
[reaction: see text] A highly stereoselective synthesis of (-)-erythrodiene starting from 4-isopropylcyclohexanone is described. The key reactions are an asymmetric methoxycarbonylation of the starting ketone and a highly diastereoselective radical cascade involving addition of a phenylthiyl radical to a terminal alkyne followed by a 1,5-hydrogen transfer and a 5-exo-cyclization.


Validation of the Acoustic Voice Quality Index in the Japanese Language.
The Acoustic Voice Quality Index (AVQI) is a multivariate construct for quantification of overall voice quality based on the analysis of continuous speech and sustained vowel. The stability and validity of the AVQI is well established in several language families. However, the Japanese language has distinct characteristics with respect to several parameters of articulatory and phonatory physiology. The aim of the study was to confirm the criterion-related concurrent validity of AVQI, as well as its responsiveness to change and diagnostic accuracy for voice assessment in the Japanese-speaking population. This is a retrospective study. A total of 336 voice recordings, which included 69 pairs of voice recordings (before and after therapeutic interventions), were eligible for the study. The auditory-perceptual judgment of overall voice quality was evaluated by five experienced raters. The concurrent validity, responsiveness to change, and diagnostic accuracy of the AVQI were estimated. The concurrent validity and responsiveness to change based on the overall voice quality was indicated by high correlation coefficients 0.828 and 0.767, respectively. Receiver operating characteristic analysis revealed an excellent diagnostic accuracy for discrimination between dysphonic and normophonic voices (area under the curve: 0.905). The best threshold level for the AVQI of 3.15 corresponded with a sensitivity of 72.5% and specificity of 95.2%, with the positive and negative likelihood ratios of 15.1 and 0.29, respectively. We demonstrated the validity of the AVQI as a tool for assessment of overall voice quality and that of voice therapy outcomes in the Japanese-speaking population.


Fe en Accion/Faith in Action: Design and implementation of a church-based randomized trial to promote physical activity and cancer screening among churchgoing Latinas.
To describe both conditions of a two-group randomized trial, one that promotes physical activity and one that promotes cancer screening, among churchgoing Latinas. The trial involves promotoras (community health workers) targeting multiple levels of the Ecological Model. This trial builds on formative and pilot research findings. Sixteen churches were randomly assigned to either the physical activity intervention or cancer screening comparison condition (approximately 27 women per church). In both conditions, promotoras from each church intervened at the individual- (e.g., beliefs), interpersonal- (e.g., social support), and environmental- (e.g., park features and access to health care) levels to affect change on target behaviors. The study's primary outcome is min/wk of moderate-to-vigorous physical activity (MVPA) at baseline and 12 and 24 months following implementation of intervention activities. We enrolled 436 Latinas (aged 18-65 years) who engaged in less than 250 min/wk of MVPA at baseline as assessed by accelerometer, attended church at least four times per month, lived near their church, and did not have a health condition that could prevent them from participating in physical activity. Participants were asked to complete measures assessing physical activity and cancer screening as well as their correlates at 12- and 24-months. Findings from the current study will address gaps in research by showing the long term effectiveness of multi-level faith-based interventions promoting physical activity and cancer screening among Latino communities.


Bilateral simultaneous endophthalmitis after immediately sequential bilateral cataract surgery.
Immediately sequential bilateral cataract surgery (ISBCS) has recently been considered effective due to faster visual rehabilitation and further improvements in quality of life. We report on the treatment course of a case of early postoperative endophthalmitis in both eyes after ISBCS. The patient was a 75-year-old woman undergoing anticancer treatment for lung cancer, who had been receiving eye drops for bilateral uveitis and secondary glaucoma since 2019. She underwent ISBCS at another hospital in 2022. In that surgery, the same instruments were used for the surgery of both eyes, without resterilization between eyes. On the night of the following day, she became aware of vision loss in both eyes, and at the examination the next day, hypopyon and corneal edema were found and she was diagnosed with bilateral postoperative endophthalmitis, and referred to our hospital. On the same day, anterior chamber irrigation, posterior capsulotomy, and vitrectomy were performed on both eyes, and the intraocular lenses were preserved. There was no recurrence of postoperative inflammation, and visual acuity was 20/30 in both eyes 6 months after vitrectomy. ISBCS is recommended to be performed with complete aseptic separation of the patient's two procedures. This important rule was not followed in this case. ISBCS should be performed in accordance with accepted protocols, such as those of the International Society of Bilateral Cataract Surgeons, the Royal College of Ophthalmologists, and the Canadian Ophthalmological Society.


Calorie controlled diet for chronic asthma.
The prevalence of asthma has increased in recent years. Epidemiological studies suggest a correlation between the onset of asthma and dietary nonallergic factors especially high calorie diet. These can be regarded as other potentially important risk factors. To observe the effect of dietary calorie reduction on chronic asthma in adults or children. We searched the Cochrane Airways Group trials register using prespecified terms. We assessed bibliographies from included studies, and contacted authors of known studies for additional information about published and unpublished trials. Date of most recent search: May 2004 Randomised-controlled trials of dietary calorie reduction were included. Three authors assessed each study independently. Disagreement was resolved by consensus. Quality assessment was performed independently. One trial of fair methodological quality with a total of 38 patients suffering from chronic asthma was included. There were significant increases in FEV(1) and FVC in the active treatment group compared with control. No data pertaining serious adverse effects were reported from the interventions. There is currently a very small amount of evidence assessing the effects of dietary interventions intended as part of a wide-ranging weight-loss programme. Whilst we are unable to recommend these strategies as concomitant interventions with drug-based therapy for the specific management of asthma, dietary interventions such as weight-loss programmes may provide benefits in specific patients. However, the impact of a calorie-controlled diet on the signs and symptoms of asthma in the general asthmatic population is yet to be established.


Efficacy and Safety of Stereotactic Body Radiation Therapy in Oligometastatic Uterine Cancer (MITO-RT2/RAD): A Large, Real-World Study in Collaboration With Italian Association of Radiation Oncology, Multicenter Italian Trials in Ovarian Cancer, and Mario Negri Gynecologic Oncology Group Groups.
This retrospective, multicenter study analyzes the efficacy and safety of stereotactic body radiation therapy in a large cohort of patients with oligometastatic/persistent/recurrent uterine cancer. Clinical and radiation therapy data from several radiation therapy centers treating patients by stereotactic body radiation therapy between March 2006 and October 2021 were collected. Objective response rate was defined as complete and partial response, and clinical benefit included objective response rate plus stable disease. Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer and Common Terminology Criteria for Adverse Events scales were used to grade toxicities. Primary endpoints were the rate of complete response to stereotactic body radiation therapy, and the 2-year actuarial local control rate "per-lesion" basis. Secondary endpoints were progression-free survival and overall survival, as well as toxicity. In the study, 157 patients with oligometastatic/persistent/recurrent uterine cancer bearing 272 lesions treated by stereotactic body radiation therapy at 14 centers were analyzed. Lymph node metastases (137, 50.4%) were prevalent, followed by parenchyma lesions (135, 49.6%). Median total dose was 35 Gy (10-75.2), in 5 fractions (range, 1-10). Complete and partial responses were 174 (64.0%), and 54 (19.9%), respectively. Stable disease was registered in 29 (10.6%), and 15 (5.5%) lesions progressed. Type of lesion (lymph node), volume (≤13.7 cc) and total dose (BED10 >59.5 Gy) were significantly associated with a higher probability of achieving complete response. Patients achieving complete response (CR) "per-lesion" basis experienced a 2-year actuarial local control rate of 92.4% versus 33.5% in lesions not achieving complete response (NCR; P < .001). Moreover, the 2-year actuarial progression-free survival rate in patients with CR was 45.4%, and patients with NCR had a 2-year rate of 17.6% (P < .001). Finally, patients who had a CR had a 2-year overall survival rate of 82.7%, compared with 56.5% for NCR patients (P <.001). Severe acute toxicity was around 2%, including one toxic death due to gastric perforation, and severe late toxicity around 4%. The efficacy of stereotactic body radiation therapy in this setting was confirmed. The low toxicity profile and the high local control rate in complete responder patients encourage the wider use of this approach.


Femtogram detection limits for biogenic amines using microbore HPLC with electrochemical detection.
Detection limits for biogenic amines, serotonin (5-HT), 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, norepinephrine, epinephrine, dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC), of 50-200 fg are reported. This was accomplished using microbore HPLC with electrochemical detection. Reproducibility at the 1-5 pg level of 1.6-8.1% was obtained. The utility of this approach is demonstrated for analysis of brain tissues of 1 microgram or less.


Cardiomyocyte-targeting exosomes from sulforaphane-treated fibroblasts affords cardioprotection in infarcted rats.
Exosomes (EXOs), tiny extracellular vesicles that facilitate cell-cell communication, are being explored as a heart failure treatment, although the features of the cell source restrict their efficacy. Fibroblasts the most prevalent non-myocyte heart cells, release poor cardioprotective EXOs. A noninvasive method for manufacturing fibroblast-derived exosomes (F-EXOs) that target cardiomyocytes and slow cardiac remodeling is expected. As a cardioprotective isothiocyanate, sulforaphane (SFN)-induced F-EXOs (SFN-F-EXOs) should recapitulate its anti-remodeling properties. Exosomes from low-dose SFN (3 μM/7 days)-treated NIH/3T3 murine cells were examined for number, size, and protein composition. Fluorescence microscopy, RT-qPCR, and western blot assessed cell size, oxidative stress, AcH4 levels, hypertrophic gene expression, and caspase-3 activation in angiotensin II (AngII)-stressed HL-1 murine cardiomyocytes 12 h-treated with various EXOs. The uptake of fluorescently-labeled EXOs was also measured in cardiomyocytes. The cardiac function of infarcted male Wistar rats intramyocardially injected with different EXOs (1·1012) was examined by echocardiography. Left ventricular infarct size, hypertrophy, and capillary density were measured. Sustained treatment of NIH/3T3 with non-toxic SFN concentration significantly enhances the release of CD81 + EXOs rich in TSG101 (Tumor susceptibility gene 101) and Hsp70 (Heat Shock Protein 70), and containing maspin, an endogenous histone deacetylase 1 inhibitor. SFN-F-EXOs counteract angiotensin II (AngII)-induced hypertrophy and apoptosis in murine HL-1 cardiomyocytes enhancing SERCA2a (sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2a) levels more effectively than F-EXOs. In stressed cardiomyocytes, SFN-F-EXOs boost AcH4 levels by 30% (p < 0.05) and significantly reduce oxidative stress more than F-EXOs. Fluorescence microscopy showed that mouse cardiomyocytes take in SFN-F-EXOs ~ threefold more than F-EXOs. Compared to vehicle-injected infarcted hearts, SFN-F-EXOs reduce hypertrophy, scar size, and improve contractility. Long-term low-dose SFN treatment of fibroblasts enhances the release of anti-remodeling cardiomyocyte-targeted F-EXOs, which effectively prevent the onset of HF. The proposed method opens a new avenue for large-scale production of cardioprotective exosomes for clinical application using allogeneic fibroblasts.


An X Chromosome Transcriptome Wide Association Study Implicates ARMCX6 in Alzheimer's Disease.
The X chromosome is often omitted in disease association studies despite containing thousands of genes which may provide insight into well-known sex differences in the risk of Alzheimer's Disease. To model the expression of X chromosome genes and evaluate their impact on Alzheimer's Disease risk in a sex-stratified manner. Using elastic net, we evaluated multiple modeling strategies in a set of 175 whole blood samples and 126 brain cortex samples, with whole genome sequencing and RNA-seq data. SNPs (MAF>0.05) within the cis-regulatory window were used to train tissue-specific models of each gene. We apply the best models in both tissues to sex-stratified summary statistics from a meta-analysis of Alzheimer's disease Genetics Consortium (ADGC) studies to identify AD-related genes on the X chromosome. Across different model parameters, sample sex, and tissue types, we modeled the expression of 217 genes (95 genes in blood and 135 genes in brain cortex). The average model R2 was 0.12 (range from 0.03 to 0.34). We also compared sex-stratified and sex-combined models on the X chromosome. We further investigated genes that escaped X chromosome inactivation (XCI) to determine if their genetic regulation patterns were distinct. We found ten genes associated with AD at p 0.05, with only ARMCX6 in female brain cortex (p = 0.008) nearing the significance threshold after adjusting for multiple testing (α = 0.002). We optimized the expression prediction of X chromosome genes, applied these models to sex-stratified AD GWAS summary statistics, and identified one putative AD risk gene, ARMCX6.


Early outcomes of lung transplantation with lung allografts from coronavirus disease 2019 (COVID-19)-positive donors.
Coronavirus disease 2019 (COVID-19) can be detected for extended periods of time with nucleic acid amplification test even after transmissibility becomes negligible. Lung allografts from COVID-19-positive donors have been used for transplantation in highly selected cases. This study aimed to clarify the early outcomes of lung transplantation with COVID-19-positive donors. The Organ Procurement and Transplantation Network/United Network for Organ Sharing database between April 2020 and June 2022 was retrospectively analyzed. In the study period, 1297 COVID-19-positive donors were identified and the lungs were transplanted from 47 donors (3.6%). Of 47 donors, 44 donors were positive for COVID-19 NAT with nasopharyngeal swabs and the other 3 were positive with bronchoalveolar lavage. The COVID-19-positive lung donors were younger than the COVID-19-negative donors (28.4 ± 11.6 years vs 35.4 ± 13.6 years, P < .001). Recipients of the COVID-19-positive lungs (n = 47) were more likely have a greater lung allocation score (57.1 ± 22.9 vs 50.5 ± 19.7, P = .057) than recipients of COVID-19-negative lungs (n = 5501). The posttransplant length of hospital stay (39.8 ± 43.6 days vs 30.6 ± 34.5 days, P = .181), need for extracorporeal membrane oxygenation support at 72 hours after transplantation (2.6% [1/38] vs 10.4% [541/5184], P = .18), and 1-year overall survival rate (85.6% vs 87.1%, P = .63) were comparable between the 2 groups. Carefully selected lung allografts from COVID-19-positive donors had comparable early posttransplant outcomes to lung allografts from COVID-19-negative donors.


Are oral medications effective in the management of acute agitation?
Current expert guidelines recommend treating agitation with oral medications instead of intramuscular medications if possible. Oral medications are sometimes believed to be inappropriate for the emergency department (ED) as they require patient cooperation and may have a slower onset of action. This review examined published literature for the efficacy of oral agents in agitation. Are oral medications effective at managing acute agitation? Structured review of PubMed of articles in which the first timepoints of evaluation were<24 hours (i.e., the typical timecourse in the ED). 11 articles included for final analysis. CONCLUSIONS/CLINICAL BOTTOM LINE: Treatment with oral medications is as effective as intramuscular medications in rapidly reducing psychotic agitation in the ED. Their use is thought to pose less risk to both patient and ED staff and is less coercive. There is little to no evidence about the use of oral medications for ED patients with extreme agitation.


A novel function for BRCA1 in crosslink repair.
In this issue of Molecular Cell, Bunting et al. (2012) provide new evidence that BRCA1 plays an important role in DNA interstrand crosslink repair that is distinct from its established function in promoting DNA end resection during homologous recombination.


Correction: An overview of atmospheric water harvesting methods, the inevitable path of the future in water supply.
[This corrects the article DOI: 10.1039/D2RA07733G.].


MR-CT multi-atlas registration guided by fully automated brain structure segmentation with CNNs.
Computed tomography (CT) is widely used to identify anomalies in brain tissues because their localization is important for diagnosis and therapy planning. Due to the insufficient soft tissue contrast of CT, the division of the brain into anatomical meaningful regions is challenging and is commonly done with magnetic resonance imaging (MRI). We propose a multi-atlas registration approach to propagate anatomical information from a standard MRI brain atlas to CT scans. This translation will enable a detailed automated reporting of brain CT exams. We utilize masks of the lateral ventricles and the brain volume of CT images as adjuvant input to guide the registration process. Besides using manual annotations to test the registration in a first step, we then verify that convolutional neural networks (CNNs) are a reliable solution for automatically segmenting structures to enhance the registration process. The registration method obtains mean Dice values of 0.92 and 0.99 in brain ventricles and parenchyma on 22 healthy test cases when using manually segmented structures as guidance. When guiding with automatically segmented structures, the mean Dice values are 0.87 and 0.98, respectively. Our registration approach is a fully automated solution to register MRI atlas images to CT scans and thus obtain detailed anatomical information. The proposed CNN segmentation method can be used to obtain masks of ventricles and brain volume which guide the registration.


Incarcerated mothers' perceptions and interpretations of their fatally maltreated children.
Fatally maltreated children are an elusive component in the complex interaction that has led to their premature deaths. Retrospective research with women imprisoned for fatal child maltreatment indicated recurring themes of maternal interpretations of their children as rejecting and developmentally abnormal, either advanced or delayed. Separations and difficulties during reunions were critical. The fatality was not a one-time event, but the exit point of a recurrent cycle of abusive interaction.


Now you see it, now you don't: variable hemineglect in a commissurotomized man.
We describe the case of a callosotomized man, D.D.V., who shows unusual neglect of stimuli in the left visual field (LVF). This is manifest in simple reaction time (RT) to stimuli flashed in the LVF and in judging whether pairs of filled circles in the LVF are of the same or different color. It may reflect strong left-hemispheric control and consequent attention restricted to the right side of space. It is not evident in simple RT when there are continuous markers in the visual fields to indicate the locations of the stimuli. In this condition, his RTs are actually faster to LVF than to right visual field (RVF) stimuli, suggesting a switch to right-hemispheric control that eliminates the hemineglect. Neglect is also not evident when D.D.V. responds by pointing to or touching the locations of the stimuli, perhaps because these responses are controlled by the dorsal rather than the ventral visual system. Despite his atypical manifestations of hemineglect, D.D.V. showed evidence of functional disconnection typical of split-brained subjects, including prolonged crossed-uncrossed different in simple reaction time, inability to match colors between visual fields, and enhanced redundancy gain in simple RT to bilateral stimuli even when the stimulus in the LVF was neglected.


[The challenge of acute kidney injury : Cross-sectional study on prevalence and costs in a university intensive care unit].
The prognosis of intensive care patients with acute kidney injury (AKI), which is associated with increased mortality is still poor. Current data on the prevalence and the resulting costs of AKI and an overview of the most common diagnoses associated with AKI in German intensive care units (ICU) are lacking. In this retrospective study all adult admissions (> 18 Jahre) in the five ICUs at the University Clinic Regensburg (in total 78 beds) from1 January 2011 to 31 December 2013 were evaluated. The ICU diagnoses commonly associated with AKI were identified using the international classification of diseases 10 (ICD 10). The length of ICU and hospital stays and AKI-associated hospital costs in the diagnosis-related groups (DRG) based reimbursement system were compared. A total of 891 ICU patients with AKI were classified according to the ICD 10 code. Acute respiratory distress syndrome (ARDS), myocardial infarction (MI) and sepsis were the three most common ICU conditions associated with AKI. A total of 1103 patients were admitted with 1 of these 3 main diagnoses and 249 (22.6 %) of these patients developed AKI. Patients with AKI had significantly longer mean ICU and hospital stays compared to patients without AKI (18.6 vs 5.1 days and 23.8 vs. 10.4 days, respectively, p < 0.001). The presence of AKI in critically ill patients with ARDS, MI and sepsis resulted in additional costs of 2,019,120.42 € at the University Hospital of Regensburg in 2013. Acute kidney injury in critically ill patients represents a significant medical and socioeconomic burden. Early recognition of patients at risk, coordinated research into novel interventions and establishment of the National Acute Kidney Injury Network for implementation of evidence-based therapies may be the next steps to decrease the incidence and severity of AKI and save costs for the national healthcare system.


Time Required for Planned and Unplanned Episodes of Care in Primary Total Joint Arthroplasty: Has Anything Changed With a Growth in Outpatient Arthroplasty?
This study sought to determine the total amount of time committed to planned and unplanned episodes of care related to primary, unilateral total joint arthroplasty (TJA), relative to a growth in outpatient TJA. All primary, unilateral TJA procedures performed over a 7-year period by a single surgeon at a single institution were retrospectively reviewed. Time dedicated to planned work was calculated over each episode of care, from surgery scheduling to 90 days postoperatively. All telephone inquiries and readmissions involving the surgeon's direct input, over the episode of care, constituted time dedicated to unplanned work. Between 2012 and 2018, as the proportion of outpatient TJAs increased, the average planned episode-of-care time per patient decreased from 412 minutes to 361 minutes. Despite a 108% increase in the total number of outpatient TJAs between 2017 and 2018 (51/432 (11.8%) to 106/555 (19.1%); P = .002), neither the average number of unplanned telephone inquiries (4.6 ± 3.8 vs 4.2 ± 3.7; P = .124), nor the mean time per patient required to respond to calls (23.1 ± 19.4 vs 21.2 ± 18 minutes, P = .135) differed. Between 2017 and 2018, the average total episode-of-care time per patient decreased from 403 minutes (376 planned + 27 unplanned) to 387 minutes (361 planned + 26 unplanned). Despite an increase in outpatient TJA, the average time required for planned and unplanned patient care remained relatively constant. The growth of outpatient TJA nationally should not trigger a change in Centers for Medicare and Medicaid Services benchmarks.


Dosing issues with non-vitamin K antagonist oral anticoagulants for the treatment of non-valvular atrial fibrillation: Why we should not underdose our patients.
Non-vitamin K antagonist oral anticoagulants (NOACs) - dabigatran, rivaroxaban, apixaban and edoxaban - are well established in terms of preventing stroke or systemic embolism in patients with non-valvular atrial fibrillation and high thromboembolism risk. When prescribed incorrectly, NOACs are associated with an increased risk of ischaemic events and bleeding. Current NOAC labels explicitly address dose adjustments according to age, body weight, renal function and concomitant treatment with P-glycoprotein inhibitors. The required dose adjustments vary significantly from molecule to molecule, thereby creating a complex dose adjustment environment. Furthermore, recommendations support assessment of individual risk using thromboembolic and bleeding risk scores. Evidence-based medicine also provides data about specific patient profiles. In particular, some patients who are at higher risk of bleeding, such as patients on polymedication, are often at higher risk of stroke. More and more patients are being treated with NOACs. The question of appropriate dosing has become important, as studies are starting to show that reduced doses are being prescribed at very high rates. Although these data have not been evaluated in light of individual risk assessments, in everyday practice, physicians are often more concerned about drug-related bleeding than about the spontaneous evolution of the disease (stroke/systemic embolism), leading to high rates of prescription of inadequately low doses. Recent results have shown that only certain risk criteria justify dose reduction. Thus, the right dose needs to be prescribed for the right patient in order to obtain, in real-life practice, the benefits of NOACs that have been demonstrated in randomized clinical trials.


Screening colonoscopy in 40- to 50-year-old first-degree relatives of patients with colorectal cancer is efficient: a controlled multicentre study.
Persons with a familial risk of colorectal cancer (CRC) account for about 25% of all CRC cases. The adenoma prevalence in relatives of CRC patients 50-60 years of age is 17-34%; data on younger individuals are scarce. Our aim was to prospectively define the adenoma prevalence in 40- to 50-year-old first-degree relatives of CRC patients compared to controls. CRC patients were identified via the regional cancer registry, and their 40- to 50-year-old first-degree relatives (risk group) were invited for screening colonoscopy. Additional probands and controls of the same age were recruited by newspaper articles and radio or television broadcastings. Using high-resolution video colonoscopy, each detected polyp was removed and histopathologically assessed. Each participant completed demographic and epidemiological questionnaires. Of 228 subjects in the risk group 36.4% had polypoid lesions compared to 20.9% of 220 controls (p<0.001). Forty-three (18.9%) subjects in the risk group had adenomas compared to 18 (8.2%) in the control group (p=0.001). High-risk adenomas (>10 mm and/or of villous type) were found in 12 persons in the risk group compared to 5 controls (not significant). In the risk group most lesions (52%) were located proximal to the sigmoid colon compared to 29% in controls. Subjects between 40-50 years with first-degree relatives with CRC demonstrate a significantly higher prevalence of adenomas than controls, with a tendency towards a more proximal location. These data support a screening colonoscopy in persons with familial risk already between 40 and 50 years.


Bringing the Vascular Lab Inside the Procedure Room.
The advantages of the vascular lab (VL) are a specialized knowledgeable team and a dedicated equipment which is transferrable to the procedure room to facilitate the procedure and gather information about the procedure endpoints. In this article, Registered Vascular Technologists (RVT) from various institutions discuss the basic and the complex ways the VL can be used inside a procedure room. The advantages of the vascular lab (VL) are a specialized knowledgeable team and dedicated equipment which is transferrable to the procedure room to facilitate the procedure and gather information about procedure endpoints. Such expertise may improve imaging guided access, enable complicated lesion crossing, and provide information about the success of the procedure. Below are the reports from practicing registered vascular technologists at the forefront of patient vascular care within institutions that have adopted the mobile intraprocedural VL idea in basic and complex real-world ways.


Size and sex allocation in monoecious woody plants.
The female size advantage hypothesis predicts that the allocation ratio of female: male reproductive effort should increase with plant size (total reproductive effort). A male height advantage hypothesis has also been proposed, based on the supposed greater advantage of height to male reproductive success in wind-pollinated plants. These ideas were tested with data for wind-pollinated, monoecious trees and shrubs which exhibit a suitably large range of sizes. Number of male inflorescences increased faster with size than did number of female inflorescences in 2 of 9 species; in the remaining 7 species there was no significant difference. The male:female ratio of inflorescence numbers increased with height in 4 of 7 species and did not change significantly in the remaining 3 species, as shown by regression. Height and size are highly correlated and so their effects could not be distinguished. The fact that many conifers place the female cones uppermost in the crown suggests that size and not height favors increased allocation to male function, as does well-established theory connecting the existence of male versus female size advantage to pollen and seed dispersal chacteristics. Regression analysis of the relation between male and female reproductive effort should be done by reduced major axis regression; ordinary least squares regression underestimates slopes; in this study opposite conclusions could be drawn from ordinary least squares and reduced major axis regressions.


ELL facilitates RNA polymerase II pause site entry and release.
Transcription is a multi-stage process that coordinates several steps within the transcription cycle including chromatin reorganization, RNA polymerase II recruitment, initiation, promoter clearance and elongation. Recent advances have identified the super elongation complex, containing the eleven-nineteen lysine-rich leukaemia (ELL) protein, as a key regulator of transcriptional elongation. Here we show that ELL has a diverse and kinetically distinct role before its assembly into the super elongation complex by stabilizing Pol II recruitment/initiation and entry into the pause site. Loss of ELL destabilizes the pre-initiation complexes and results in disruption of early elongation and promoter proximal chromatin structure before recruitment of AFF4 and other super elongation complex components. These changes result in significantly reduced transcriptional activation of rapidly induced genes. Thus, ELL has an early and essential role during rapid high-amplitude gene expression that is required for both Pol II pause site entry and release.


Induction of Secondary Carotenogenesis in New Halophile Microalgae from the Genus Dunaliella (Chlorophyceae).
We report on the effects of high light irradiance (480 µmol quanta/(m(2)·s)) and salinity (160 and 200 g/liter NaCl) on culture growth as well as on cell lipid pigment and fatty acid (FA) composition in three novel strains of halophile microalga from the genus Dunaliella. Based on the ITS1-5.8S rRNA-ITS2 sequence and on the capability of accumulation of secondary (uncoupled from the photosynthetic apparatus) β-carotene, the strains Dunaliella sp. BS1 and BS2 were identified as D. salina and Dunaliella sp. R5 as D. viridis. Under conditions optimal for growth, chlorophylls and primary carotenoids (mainly lutein) dominated the pigment profile of all investigated strains. The main FA were represented by unsaturated C18 FA typical of thylakoid membrane structural lipids. In all studied cells, stressors caused a decline in chlorophylls and an increase in unsaturated C16 and C18 FA associated with reserve lipids. The carotenogenic species D. salina demonstrated 10-fold increase in carotenoids accompanied by a decline in lutein and a drastic increase in β-carotene (up to 75% of total carotenoids). In D. viridis, only 1.5-fold increase in carotenoid content took place, the ratio of major carotenoids remaining essentially unchanged. The role of the carotenogenic response in mechanisms of protection against photooxidative damage is discussed in view of halophile microalgae stress tolerance and application of the new Dunaliella strains for biotechnological production of β-carotene.


Volume-Function Analysis (LiMAx Test) in Patients with HCC and Cirrhosis Undergoing TACE-A Feasibility Study.
Transarterial chemoembolization (TACE) is an important therapy for hepatocellular carcinoma (HCC) in cirrhosis. In particular in advanced cirrhosis, post-TACE hepatic failure liver (PTHF) failure may develop. Currently, there is no standardization for the periinterventional risk assessment. The liver maximum capacity (LiMAx) test assesses the functional liver capacity, but has not been investigated in this setting. The aim of this study was to prospectively evaluate periinterventional LiMAx and CT volumetry measurements in patients with cirrhosis and HCC undergoing repetitive TACE. From 06/2016 to 11/2017, eleven patients with HCC and cirrhosis undergoing TACE were included. LiMAx measurements (n = 42) were conducted before and after each TACE. Laboratory parameters were correlated with the volume-function data. The median LiMAx levels before (276 ± 166 µg/kg/h) were slightly reduced after TACE (251 ± 122 µg/kg/h; p = 0.08). This corresponded to a median drop of 7.1%. Notably, there was a significant correlation between LiMAx levels before TACE and bilirubin (but not albumin nor albumin-bilirubin [ALBI] score) increase after TACE (p = 0.02, k = 0.56). Furthermore, a significantly higher increase in bilirubin in patients with LiMAx ≤ 150 µg/kg/h was observed (p = 0.011). LiMAx levels at different time points in single patients were similar (p = 0.2). In our prospective pilot study in patients with HCC and cirrhosis undergoing multiple TACE, robust and reliable LiMAx measurements were demonstrated. Lower LiMAx levels before TACE were associated with surrogate markers (bilirubin) of liver failure after TACE. Specific subgroups at high risk of PTHF should be investigated. This might facilitate the future development of strategies to prevent occurrence of PTHF.


High prevalence of Cryptococcus neoformans var. gattii in tropical and subtropical regions.
The global distribution of 96 clinical isolates of Cryptococcus neoformans var. gattii among a total of 725 C. neoformans isolates was investigated. While C. neoformans var. gattii was not found among isolates obtained from Europe (excluding the United Kingdom) and Japan; and it was found only infrequently in North America (excluding Southern California), the United Kingdom, New Zealand, and Argentina; the variety was prevalent in regions with tropical to subtropical climates such as South East Asia, Southern California, Brazil, Mid to South Pacific, and the central part of Africa. Among the isolates of C. neoformans var. gattii in our collection, serotype B predominanted (77 of 96), and the serotype C isolates were almost exclusively from North America.


Insulin, transforming growth factors, and substrates modulate growth of guinea pig pancreatic duct cells in vitro.
Little is known of the physiological mechanisms that control cellular renewal in the pancreatic excretory duct system. This study investigated the effects of potential regulatory substances on the growth of cultured guinea pig pancreatic duct epithelial monolayers. Pancreatic duct explants were cultured for 3 days on plastic and on permeable filters in the presence and absence of different substances. Growth of epithelial monolayers from these explants was measured by 5-bromo-2'-deoxyuridine incorporation and morphometric procedures. Epidermal growth factor and insulin both enhanced monolayer growth and together had an additive effect. Transforming growth factor alpha enhanced and transforming growth factor beta inhibited growth, whereas glucagon, somatostatin, pancreatic polypeptide, secretin, cerulein, bombesin, and dexamethasone had no significant effects. Monolayer growth on type 1 collagen-coated filters was enhanced when compared with that of monolayers grown on tissue culture plastic. Cell growth from explants on filters coated with type IV collagen and fibronectin was comparable with that on plastic, whereas growth on Matrigel- or laminin-coated filters was reduced. Insulin, transforming growth factors, and substrate components modulate growth of pancreatic duct epithelial cells in vitro, suggesting that they are important regulators of cell division in the excretory duct system of the intact pancreas.


Facile preparation of pH-sensitive chitosan microspheres for delivery of curcumin; characterization, drug release kinetics and evaluation of anticancer activity.
Curcumin (CUR) is a lowly water-soluble natural polyphenol with chemopreventive and chemotherapeutic activities. Hence, to achieve the system with good CUR loading ability, porous MIL-88 (Fe) was prepared in the presence of the presynthesized graphene quantum dots (GQDs) (GQDs@MIL-88 (Fe)). In the following, CUR loaded in the fabricated GQDs@MIL-88 (Fe) nanohybrid. The characterization techniques; Fourier transform infrared (FT-IR), X-ray powder diffraction (XRD), scanning electron microscope (SEM), photoluminescence (PL), and Brunauer-Emmett-Teller (BET) analysis showed success in the synthesis of GQDs@MIL-88 (Fe). Moreover, the FT-IR analysis displayed the loading of CUR and the formation of CUR@GQDs@MIL-88(Fe). Chitosan (CS) was used as a green coating to enhance the biocompatibility of the prepared system (CS/CUR@GQDs@MIL-88(Fe). The fabricated microspheres showed pH-sensitive swelling behavior and released 38.3% of CUR in pH 5.0 which is better fitted with the First-order kinetic model (R2 = 0.9726). In comparison with CUR@GQDs@MIL-88(Fe), the MTT and DAPI assay exhibited less toxic effect for CS/CUR@GQDs@MIL-88(Fe) against MDA-MB 231 cells. Moreover, the safety of the CS/CUR@GQDs@MIL-88(Fe) confirmed after incubation against MCF 10A as a model of the normal cell line. The results conveyed a new concept that the CS/CUR@GQDs@MIL-88(Fe) is a potential candidate for using as a biocompatible carrier with controlled drug delivery ability.


Extraction of DNA from Skeletal Remains.
Acquisition of DNA from skeletal remains can be a delicate process. With the advent of improved extraction buffers that provide complete demineralization of the osseous materials, extraction of total genomic DNA from nearly any skeletal element is possible. This chapter describes both traditional organic and more newly developed inorganic extraction methods for fresh and dried skeletal remains.


Methimazole Treatment and Risk of Acute Pancreatitis: A Population-based Cohort Study.
A warning has been recently issued by the European Medicine Agency (EMA) regarding a potential increased risk of acute pancreatitis (AP) in methimazole (MMI) users. To investigate the association between MMI and the diagnosis of AP in a population-based study. A retrospective analysis of administrative health databases was conducted (2013-2018). Relevant data were obtained from: (1) inhabitants registry, (2) hospital discharge records (ICD-9-CM 577.0), and (3) drug claims registry (ATC H03BB02). We evaluated AP risk in MMI users in 18 months of treatment, stratifying results by trimester. Poisson regression was used to estimate the age- and sex-adjusted rate ratios (RR), and the relative 95% confidence intervals (CI), comparing rates of AP between MMI users and nonusers. The absolute risk of AP in MMI users was also calculated. A total of 23 087 new users of MMI were identified. Among them, 61 hospitalizations occurred during the study period. An increase in AP risk was evident during the first 3 trimesters of therapy (RR 3.40 [95% CI: 2.12-5.48]; RR 2.40 [95% CI: 1.36-4.23]; RR 2.80 [95% CI: 1.66-4.73]), but disappeared thereafter. The AP absolute risk in MMI users during the first 18 months of treatment was less than 0.4% in all sex and age classes. Our results support the EMA warning, suggesting an increased risk of AP associated with MMI use. However, such an increase seems limited to the first months of MMI treatment. Moreover, in absolute terms, the probability of AP is low among patients, well below 1%.


Pediatric Gliosarcoma With and Without Neurofibromatosis Type 1: A Whole-exome Comparison of 2 Patients.
Gliosarcoma is rare among pediatric patients and among individuals with Neurofibromatosis Type 1 (NF1). Here we compare 2 pediatric gliosarcoma patients, one of whom has NF1. We performed whole-exome sequencing, methylation, and copy number analysis on tumor and blood for both patients. Whole-exome sequencing showed higher mutational burden in the tumor of the patient without NF1. Copy number analysis showed differences in chromosomal losses/gains between the tumors. Neither tumor showed O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation. The NF1 patient survived without progression while the other expired. This is the first reported case of gliosarcoma in a child with NF1.


Controlled Synthesis of a Vacancy-Defect Single-Atom Catalyst for Boosting CO2 Electroreduction.
The reaction of precursors containing both nitrogen and oxygen atoms with NiII under 500 °C can generate a N/O mixing coordinated Ni-N3 O single-atom catalyst (SAC) in which the oxygen atom can be gradually removed under high temperature due to the weaker Ni-O interaction, resulting in a vacancy-defect Ni-N3 -V SAC at Ni site under 800 °C. For the reaction of NiII with the precursor simply containing nitrogen atoms, only a no-vacancy-defect Ni-N4 SAC was obtained. Experimental and DFT calculations reveal that the presence of a vacancy-defect in Ni-N3 -V SAC can dramatically boost the electrocatalytic activity for CO2 reduction, with extremely high CO2 reduction current density of 65 mA cm-2 and high Faradaic efficiency over 90 % at -0.9 V vs. RHE, as well as a record high turnover frequency of 1.35×105 h-1 , much higher than those of Ni-N4 SAC, and being one of the best reported electrocatalysts for CO2 -to-CO conversion to date.


Disease modification in epilepsy: from animal models to clinical applications.
Several relevant animal models of epileptogenesis and biomarkers have emerged for evaluating the antiepileptogenic potential of an investigational drug. Although several promising candidate compounds and approaches have been identified in these preclinical models, no treatment has yet successfully navigated the path from preclinical efficacy to clinical validation. Until such an agent can move from preclinical proof of concept to clinical success, the need remains to continually develop and optimize preclinical models and clinical trial design in an effort to guide potential clinical investigations. This review describes several available models of disease modification and/or epileptogenesis, preclinical studies in these models and potential biomarkers useful for evaluating the efficacy of a potential therapeutic agent in the preclinical setting. The results that emerge from such efforts may then guide the clinical evaluation of a candidate compound. This review discusses some of the known limitations and hurdles to moving compounds found effective in these models to clinical practice, in the hope that knowledge of this information will facilitate the design and conduct of clinical studies and effectively facilitate the identification of a first-in-class disease-modifying or antiepileptogenic agent.


Complexation behaviour and removal of organic-Cr(III) complexes from the environment: A review.
Chromium (Cr) is mainly found in the form of organic-Cr(III) complexes in the natural environment and industrial waste. The widespread existence of composite contaminants composed of organic matter (OM) and Cr pose a serious ecological threat, and its potential interaction and removal need to be further summarised. Organic ligands, such as carbohydrates, nitrogen compounds, phenolic compounds, humus substances (HS), and low molecular weight organic acids (LMWOAs), play an important role in governing the speciation, mobility, and absorption and desorption of Cr in the environment. Moreover, growing evidence indicates that oxygen-containing functional groups (e.g., carboxyl, hydroxyl, and phosphate) are closely related to the complexation of Cr(III). Advanced oxidation processes (AOPs) are efficient and widely applicable technologies. However, the re-complexation of oxidation intermediates with Cr(III) and the formation and accumulation of much more toxic Cr(VI) species hinder the possible utilisation of AOPs. In this paper, the sources and harmful effects of organic-Cr(III) complexes are reported in detail. The complexation behaviour and structure of the organic-Cr(III) complexes are also described. Subsequently, the application of AOPs in the decomplexation and degradation of organic-Cr(III) complexes is summarised. This review can be helpful for developing technologies that are more efficient for organic-Cr(III) complex removal and establishing the scientific background for reducing Cr discharge Cr into the environment.


Using low frequency full bottle diamagnetic screening to study collectible wine.
A low frequency ν < 30 MHz spectrometer capable of noninvasively and nondestructively screening the diamagnetic properties of full intact bottles of wine is described, and along with principal component analysis, used to compare and contrast sealed bottles of wine. The sensitivity of this approach to various ionic and molecular wine solutes is established by analyzing standard solutions. The successful application of this full bottle method to a library of collectible wine is discussed and suggests that the method can be used to identify counterfeit wine without violating the bottle.


Improving biomarker list stability by integration of biological knowledge in the learning process.
The identification of robust lists of molecular biomarkers related to a disease is a fundamental step for early diagnosis and treatment. However, methodologies for biomarker discovery using microarray data often provide results with limited overlap. It has been suggested that one reason for these inconsistencies may be that in complex diseases, such as cancer, multiple genes belonging to one or more physiological pathways are associated with the outcomes. Thus, a possible approach to improve list stability is to integrate biological information from genomic databases in the learning process; however, a comprehensive assessment based on different types of biological information is still lacking in the literature. In this work we have compared the effect of using different biological information in the learning process like functional annotations, protein-protein interactions and expression correlation among genes. Biological knowledge has been codified by means of gene similarity matrices and expression data linearly transformed in such a way that the more similar two features are, the more closely they are mapped. Two semantic similarity matrices, based on Biological Process and Molecular Function Gene Ontology annotation, and geodesic distance applied on protein-protein interaction networks, are the best performers in improving list stability maintaining almost equal prediction accuracy. The performed analysis supports the idea that when some features are strongly correlated to each other, for example because are close in the protein-protein interaction network, then they might have similar importance and are equally relevant for the task at hand. Obtained results can be a starting point for additional experiments on combining similarity matrices in order to obtain even more stable lists of biomarkers. The implementation of the classification algorithm is available at the link: http://www.math.unipd.it/~dasan/biomarkers.html.


Regulation of ryanodine receptors by sphingosylphosphorylcholine: involvement of both calmodulin-dependent and -independent mechanisms.
Sphingosylphosphorylcholine (SPC), a lipid mediator with putative second messenger functions, has been reported to regulate ryanodine receptors (RyRs), Ca2+ channels of the sarco/endoplasmic reticulum. RyRs are also regulated by the ubiquitous Ca2+ sensor calmodulin (CaM), and we have previously shown that SPC disrupts the complex of CaM and the peptide corresponding to the CaM-binding domain of the skeletal muscle Ca2+ release channel (RyR1). Here we report that SPC also displaces Ca2+-bound CaM from the intact RyR1, which we hypothesized might lead to channel activation by relieving the negative feedback Ca2+CaM exerts on the channel. We could not demonstrate such channel activation as we have found that SPC has a direct, CaM-independent inhibitory effect on channel activity, confirmed by both single channel measurements and [3H]ryanodine binding assays. In the presence of Ca2+CaM, however, the addition of SPC did not reduce [3H]ryanodine binding, which we could explain by assuming that the direct inhibitory action of the sphingolipid was negated by the simultaneous displacement of inhibitory Ca2+CaM. Additional experiments revealed that RyRs are unlikely to be responsible for SPC-elicited Ca2+ release from brain microsomes, and that SPC does not exert detergent-like effects on sarcoplasmic reticulum vesicles. We conclude that regulation of RyRs by SPC involves both CaM-dependent and -independent mechanisms, thus, the sphingolipid might play a physiological role in RyR regulation, but channel activation previously attributed to SPC is unlikely.