Cells adapt their shapes with their experienced stiffness: A geometrical approach to differentiation.
The elasticity-directed differentiation of mesenchymal stem cells has been widely studied since mid 2000s. Over nearly linear-elastic materials the differentiation of the stem cells are shown to be related to the Young's modulus of the substrate. While it is found that constraining the stem cells to some pre-patterned shape and size affects their differentiation, it has been also well recognized that the cell morphology intimately relates to its type and functionality such that the cell morphology has been accepted as an indicator for the differentiation path. This paper conjectures the importance of geometry in differentiation and claims that the elasticity indicator for differentiation is indeed "the stiffness" which contains not only the elasticity coefficient but also the geometrical information. An elasticity model is derived for a singular cell with different shapes over a thick substrate, which almost resembles the condition for most of the tests with sparse distribution of the cells. Analysis of principle shapes such as square, rectangle, and hexagon (with and without dendrites) suggest that the larger the aspect ratio i.e. the further the shape to the roundness, the larger the substrate stiffness experienced by the cells. By moving towards a more round shape such as a hexagon with or without dendrites the substrate stiffness falls off rapidly. Then by including the stiffness of the cell body itself we arrive at a more important finding; the cells at free culture condition prefer the shape which best equalizes their experienced stiffness of the substrate and that of their own body. The body-to-substrate stiffness ratio, hence, explains why a slender rectangle is the preferred shape at myogenic differentiation range and a hexagon with dendrites is the preferred one at neurogenic range.


Differential tissue specific expression of Kif23 alternative transcripts in mice with the human mutation causing congenital dyserythropoietic anemia type III.
Kinesin Family Member 23 (KIF23), a cell cycle regulator, has a key task in cytokinesis. KIF23 over-expression in cancer has been associated with tumor growth, progression, and poor prognosis, indicating a potential to be a cancer biomarker. A mutation in KIF23 (c.2747C > G, p.P916R) was shown to cause congenital dyserythropoietic anemia, type III (CDA III). To-date, fifteen KIF23 transcripts have been annotated, but their expression is poorly investigated. We hypothesized that tissue specific expression of a particular transcript can be critical for CDA III phenotype. In this study, we quantified expression of alternative Kif23 transcripts in a mouse model with human KIF23 mutation and investigated its association with a regulator of alternative splicing, serine/arginine-rich splicing factor 3 (Srsf3). We confirmed presence of an additional exon 8 in both human and mouse KIF23 transcripts. A transcript lacking exons 17 and 18 was ubiquitously expressed in mice while other isoforms were common in human tissues however in bone marrow of knock-in mice a transcript without exon 18 was prevalent as it was in bone marrow of a CDA III patient. We conclude that the possibility that the tissue specific expression of KIF23 alternative transcripts influence the CDA III phenotype cannot be neglected.


The direct regulation of Aalbdsx on AalVgR is indispensable for ovarian development in Aedes albopictus.
Aedes albopictus is an important vector with an extensive worldwide distribution. Only female mosquitoes play a significant role in the transmission of pathogens. Doublesex (dsx) is a central nexus gene in the insect somatic sex determination hierarchy. In this study, we characterized the full-length sex-specific splicing forms of the Ae. albopictus dsx (Aalbdsx) gene. Then, we identified 15 direct target genes of DSX in adult females using digital gene expression combined with quantitative real-time polymerase chain reaction (qPCR) by performing a chromatin immunoprecipitation (ChIP) assay with specific DSX antibodies. Knockdown of Aalbdsx suppressed ovarian development and decreased the transcript levels of the Aalbdsx target vitellogenin receptor (VgR) gene, whereas vitellogenin (Vg) expression showed an increase in the fat body. Genes in the major Vg regulatory pathway were also up-regulated. Our results suggest that both Vg and VgR are direct target genes of Aalbdsx and that direct regulation of Aalbdsx on VgR is indispensable for ovarian development in Ae. albopictus, which not only provides a reference for the further elucidation of the evolutionarily conserved role of dsx in Ae. albopictus sexual differentiation but also reveals potential molecular targets for application to the development of sterile male mosquitoes to be released for vector control. © 2020 Society of Chemical Industry.


Mechanisms in simulated torsade de pointes.
A mechanism of torsade de pointes consisting of moving sites of reentrant excitation has been proposed on the basis of findings with a computer model. Substantial additions to that mechanism are now proposed based on further studies with the same model. The model simulated propagation, cycle length-dependent recovery of excitability, and slow propagation during incomplete recovery. Regions of relatively short and long recovery were assigned because of evidence of regional prolongation of recovery in long QT syndromes in which torsade de pointes is frequent. As previously reported, premature excitation in the short recovery region initially propagated independently, then entered the long recovery region and reentered the short recovery region distal to the site of origin. Reentrant excitation initiated a similar series of events, and serial reentry at systematically changing locations resulted in changing patterns of excitation compatible with the changing QRS waveform in torsade de pointes. Episodes terminated when reentrant excitation reached the end of unclosed short recovery paths, collided in closed paths, or encountered refractoriness in the presence of nonuniform short recovery. In this study, it was shown that excitation preceding reentry had important effects on the mechanism. These included reversal of the direction of serial reentry, bidirectional serial reentry, reentry at multiple sites from the same parent conditions, and occurrence of reentry without the requirement of slow propagation. Evidence for a Doppler shift of cycle lengths in regions from which serial reentry was receding or approaching was obtained. Sustained serial reentry was also demonstrated and is a possible mechanism for polymorphic ventricular tachycardia. Findings further define a possible mechanism of torsade de pointes.


Interactions of different inhibitors with active-site aspartyl residues of HIV-1 protease and possible relevance to pepsin.
The importance of the active site region aspartyl residues 25 and 29 of the mature HIV-1 protease (PR) for the binding of five clinical and three experimental protease inhibitors [symmetric cyclic urea inhibitor DMP323, nonhydrolyzable substrate analog (RPB) and the generic aspartic protease inhibitor acetyl-pepstatin (Ac-PEP)] was assessed by differential scanning calorimetry. DeltaT(m) values, defined as the difference in T(m) for a given protein in the presence and absence of inhibitor, for PR with DRV, ATV, SQV, RTV, APV, DMP323, RPB, and Ac-PEP are 22.4, 20.8, 19.3, 15.6, 14.3, 14.7, 8.7, and 6.5 degrees C, respectively. Binding of APV and Ac-PEP is most sensitive to the D25N mutation, as shown by DeltaT(m) ratios [DeltaT(m)(PR)/DeltaT(m)(PR(D25N))] of 35.8 and 16.3, respectively, whereas binding of DMP323 and RPB (DeltaT(m) ratios of 1-2) is least affected. Binding of the substrate-like inhibitors RPB and Ac-PEP is nearly abolished (DeltaT(m)(PR)/DeltaT(m)(PR(D29N)) > or = 44) by the D29N mutation, whereas this mutation only moderately affects binding of the smaller inhibitors (DeltaT(m) ratios of 1.4-2.2). Of the nine FDA-approved clinical HIV-1 protease inhibitors screened, APV, RTV, and DRV competitively inhibit porcine pepsin with K(i) values of 0.3, 0.6, and 2.14 microM, respectively. DSC results were consistent with this relatively weak binding of APV (DeltaT(m) 2.7 degrees C) compared with the tight binding of Ac-PEP (DeltaT(m) > or = 17 degrees C). Comparison of superimposed structures of the PR/APV complex with those of PR/Ac-PEP and pepsin/pepstatin A complexes suggests a role for Asp215, Asp32, and Ser219 in pepsin, equivalent to Asp25, Asp25', and Asp29 in PR in the binding and stabilization of the pepsin/APV complex.


A comparison of the effects of the three isomers of dinitrobenzene on the testis in the rat.
Sexually mature Alpk/AP (Wistar derived) rats were killed 5 days after a single oral dose of 50 mg/kg of the 1,2-, 1,3-, or 1,4-isomers of dinitrobenzene. Testis weight reductions accompanied by testicular lesions were observed in the animals dosed with 1,3-dinitrobenzene (1,3-DNB) while the 1,2- and 1,4-isomers were without effect on the testis. However, 1,4-dinitrobenzene, but not 1,2-dinitrobenzene, was of a potency similar to that of 1,3-DNB in producing cyanosis and splenic enlargement in these animals, indicating that different mechanisms are probably responsible for these two toxic effects. In a subsequent study the pathogenesis of the testicular damage resulting from a single oral dose of 5, 10, 15, or 25 mg 1,3-DNB/kg was studied in sexually mature rats. Animals were killed 6, 12, 24, 48, and 96 hr after dosing and a detailed histopathological examination of the testes and selected tissues was made. At 12 hr after a single oral dose of 25 mg/kg, 1,3-DNB produced testicular lesions limited to Stages VIII to XI of the spermatogenic cycle. By 24 hr widespread Sertoli cell damage was evident and in some tubules was associated with degeneration of primary spermatocytes. Ultrastructural examination at this time confirmed that there were effects on Sertoli cells in the absence of germ cell damage. Similar effects were seen 48 hr after a single oral dose of 15 mg 1,3-DNB/kg. Doses of 5 or 10 mg 1,3-DNB/kg were without effect on the testis. The Sertoli cell is implicated as the prime target for the toxic action of 1,3-DNB with germ cell damage a secondary event.


Construction of a bacteriophage-derived recombinase system in Bacillus licheniformis for gene deletion.
Bacillus licheniformis and its related strains have found extensive applications in diverse industries, agriculture, and medicine. However, the current breeding methods for this strain primarily rely on natural screening and traditional mutagenesis. The limited availability of efficient genetic engineering tools, particularly recombination techniques, has hindered further advancements in its applications. In this study, we conducted a comprehensive investigation to identify and characterize a recombinase, RecT, derived from a Bacillus phage. Remarkably, the recombinase exhibited a 105-fold enhancement in the recombination efficiency of the strain. To facilitate genome editing, we developed a system based on the conditional expression of RecT using a rhamnose-inducible promoter (Prha). The efficacy of this system was evaluated by deleting the amyL gene, which encodes an α-amylase. Our findings revealed that the induction time and concentration of rhamnose, along with the generation time of the strain, significantly influenced the editing efficiency. Optimal conditions for genome editing were determined as follows: the wild-type strain was initially transformed with the genome editing plasmid, followed by cultivation and induction with 1.5% rhamnose for 8 h. Subsequently, the strain was further cultured for an additional 24 h, equivalent to approximately three generations. Consequently, the recombination efficiency reached an impressive 16.67%. This study represents a significant advancement in enhancing the recombination efficiency of B. licheniformis through the utilization of a RecT-based recombination system. Moreover, it provides a highly effective genome editing tool for genetic engineering applications in this strain.


Maternal cadmium exposure induces mt2 and smtB mRNA expression in zebrafish (Danio rerio) females and their offspring.
The present study aimed to identify the effects of maternal cadmium (Cd(2+)) exposure on the mRNA expression of mt2 (metallothionein-2) and smtB (similar to metallothionein-B) in female zebrafish (Danio rerio) and their offspring (F1 larvae). Zebrafish females were exposed to 0, 8.9, 17.8, and 35.6 μM Cd(2+) for 72 h, and their ovaries and F1 larvae were collected to measure their Cd(2+) contents and their smtB and mt2 mRNA expression. Cd(2+) contents and the mRNA expression of smtB and mt2 in F1 larvae all showed positive correlations with the maternal Cd(2+) treatment dose. The mt2 was 1.9- to 3.4-fold higher than smtB in F1 larvae. Furthermore, F1 larvae had noticeably enhanced Cd(2+) tolerance after maternal Cd(2+) treatment. These results demonstrate that maternal Cd(2+) was transferred to larval fish and induced mt2 and smtB mRNA expression to protect larva against the impacts of Cd(2+). In female ovaries, mt2 expression showed a noticeable increase after exposure to a metal environment, while smtB did not show exactly the same effect. The study can only conclude that smtB might have a much different role other than just protecting against the impacts of metals.


Late reoperation for systemic atrioventricular valve regurgitation after repair of congenital heart defects.
Since 1979, 17 infants and children have undergone reoperation for systemic atrioventricular (AV) valve regurgitation 6 weeks to 7 years after repair of congenital heart defects. Prior operations were repair of incomplete or complete AV canal (14 patients), Mustard repair of complex transposition of the great arteries including ventricular septal defect closure (2 patients), or first-stage operation for hypoplastic left heart (1 patient). Age ranged from 6 months to 11 years. In 12 of the 17 patients (10, AV canal; 1, transposition; 1, hypoplastic left heart), valve reconstruction was possible. Operative techniques included a combination of septal cleft approximation, leaflet resection, commissural annuloplasty, or ring annuloplasty. There were no operative deaths, and there were no reoperations in the repair group. The condition of these patients has improved. Follow-up ranges from 1 month to 9 years (mean follow-up, 4.1 years). Five of the 17 patients (4, AV canal; 1, transposition) underwent valve replacement. There were no operative deaths. Follow-up ranges from 3 to 8 years. Three patients later underwent re-replacement of the prosthetic valve; there was 1 late death. The condition of all 4 survivors is improved. Substantial AV valve regurgitation can occur months or years after repair of congenital heart defects. A combination of reconstructive techniques may be useful in preserving native valve function and avoiding systemic AV valve replacement.


Construction of an automatic score for the evaluation of speech disorders among patients treated for a cancer of the oral cavity or the oropharynx: The Carcinologic Speech Severity Index.
Speech disorders impact quality of life for patients treated with oral cavity and oropharynx cancers. However, there is a lack of uniform and applicable methods for measuring the impact on speech production after treatment in this tumor location. The objective of this work is to (1) model an automatic severity index of speech applicable in clinical practice, that is equivalent or superior to a severity score obtained by human listeners, via several acoustics parameters extracted (a) directly from speech signal and (b) resulting from speech processing and (2) derive an automatic speech intelligibility classification (i.e., mild, moderate, severe) to predict speech disability and handicap by combining the listener comprehension score with self-reported quality of life related to speech. Eighty-seven patients treated for cancer of the oral cavity or the oropharynx and 35 controls performed different tasks of speech production and completed questionnaires on speech-related quality of life. The audio recordings were then evaluated by human perception and automatic speech processing. Then, a score was developed through a classic logistic regression model allowing description of the severity of patients' speech disorders. Among the group of parameters subject to extraction from automatic processing of the speech signal, six were retained, producing a correlation at 0.87 with the perceptual reference score, 0.77 with the comprehension score, and 0.5 with speech-related quality of life. The parameters that contributed the most are based on automatic speech recognition systems. These are mainly the automatic average normalized likelihood score on a text reading task and the score of cumulative rankings on pseudowords. The reduced automatic YC2SI is modeled in this way: YC2SIp = 11.48726 + (1.52926 × Xaveraged normalized likelihood reading ) + (-1.94e-06 × Xscore of cumulative ranks pseudowords ). Automatic processing of speech makes it possible to arrive at valid, reliable, and reproducible parameters able to serve as references in the framework of follow-up of patients treated for cancer of the oral cavity or the oropharynx.


NPM1-mutated acute myeloid leukemia of monocytic or myeloid origin exhibit distinct immunophenotypes.
Acute myeloid leukemia with mutated nucleophosmin (NPM1m+AML) is a heterogeneous entity. We investigated whether NPM1m+AML with monocytic or myeloid differentiation have distinct immunophenotype. The study included 160 NPM1m+AMLpatients and 178 AML patients without NPM1 mutation and recurrent cytogenetic abnormality (NPM1wt-AML). We analyzed the immunophenotype by flow cytometry. NPM1 mutation was detected by PCR. Compared with NPM1wt-AML patients, NPM1m+AML patients showed higher positive rates of CD33 and CD9 and lower positive rates of CD34, HLA-DR, CD7, CD15 and CD117 (all P<0.05). HLA-DR, CD64, CD14, CD11b, CD15, CD4, CD9 and CD10 were higher (P<0.001) and CD117 was lower (P<0.01) in monocytic NPM1m+AML compared with myeloid NPM1m+AML. Similar rates of lymphoid antigen (CD19, CD2, and CD7) and myeloid antigen (CD13, CD33) positivity were detected in monocytic and myeloid NPM1m+AML. Compared with NPM1wt-AML, CD34 expression was lower both in myeloid and monocytic NPM1m+AML subgroups, although HLA-DR was lower in NPM1m+AML compared with NPM1wt-AML only in myeloid subgroup. Comparisons of NPM1m+AML and NPM1wt-AML showed no differences in monocyte-associated markers such as CD14 and CD11b in myeloid and monocytic subgroup. Myeloid NPM1m+AML correlated with the female gender (P=0.001), lower WBC counts (P=0.04) and higher WT1 transcripts (P=0.006) compared with monocytic NPM1m+AML.These results suggested monocytic and myeloid-derived NPM1m+AML exhibit distinct immunophenotypes.


Expectation modulates neural representations of valence throughout the human brain.
The brain's sensitivity to unexpected gains or losses plays an important role in our ability to learn new behaviors (Rescorla and Wagner, 1972; Sutton and Barto, 1990). Recent work suggests that gains and losses are ubiquitously encoded throughout the human brain (Vickery et al., 2011), however, the extent to which reward expectation modulates these valence representations is not known. To address this question, we analyzed recordings from 4306 intracranially implanted electrodes in 39 neurosurgical patients as they performed a two-alternative probability learning task. Using high-frequency activity (HFA, 70-200 Hz) as an indicator of local firing rates, we found that expectation modulated reward-related neural activity in widespread brain regions, including regions that receive sparse inputs from midbrain dopaminergic neurons. The strength of unexpected gain signals predicted subjects' abilities to encode stimulus-reward associations. Thus, neural signals that are functionally related to learning are widely distributed throughout the human brain.


Patient preference regarding first-trimester aneuploidy risk assessment.
We assessed patient views regarding disclosure of first-trimester Down syndrome risk assessment results compared with withholding results until a single, more accurate second-trimester screening result is available. A prospective, institutional review board-approved, voluntary, anonymous survey was presented to patients with singleton pregnancies who were undergoing first-trimester nuchal translucency and biochemical screening at our institution. Options included immediate (sequential testing) or later disclosure (integrated testing). Descriptions and a comparison of the options were included in the survey. One hundred one women completed the questionnaires; 69.3% of the patients preferred sequential testing, compared with 30.7% of the patients who were either unsure or preferred integrated testing. Older patients and those patients with better background knowledge of screening tests preferred earlier disclosure of screening results (P = .035 and P = .026, respectively). Patients who preferred earlier disclosure also preferred termination of a Down syndrome pregnancy (P = .013). Older patients and those patients with a better understanding of screening tests preferred immediate disclosure of first-trimester risk assessment results; these women were more likely to prefer to terminate an affected pregnancy.


Echocardiographic diagnosis of pneumopericardium and hydropneumopericardium.
Pneumopericardium and hydropneumopericardium are rare conditions in which hemodynamic compromise is common. Loss of the M-mode and two-dimensional image coincident with the systolic phase of each cardiac cycle and swirling echogenic bubbles within the pericardial space produce a distinctive M-mode and two-dimensional echocardiographic appearance. This provides an additional method for prompt diagnosis.


Kinetic study of the reactions between chloramine disinfectants and hydrogen peroxide: temperature dependence and reaction mechanism.
The temperature-dependent kinetics for the reaction between hydrogen peroxide and chloramine water disinfectants (NH2Cl, NHCl2, and NCl3) have been determined using stopped flow-UV/Vis spectrophotometry. Rate constants for the mono- and dichloramine-peroxide reaction were on the order of 10(-2)M(-1)s(-1) and 10(-5)M(-1)s(-1), respectively. The reaction of trichloramine with peroxide was negligibly slow compared to its thermal and photolytically-induced decomposition. Arrhenius expressions of ln(kH2O2-NH2Cl)=(17.3±1.5)-(51500±3700)/RT and ln(kH2O2-NHCl2)=(18.2±1.9)-(75800±5100)/RT were obtained for the mono- and dichloramine peroxide reaction over the temperature ranges 11.4-37.9 and 35.0-55.0°C, respectively. Both monochloramine and hydrogen peroxide were first-order in the rate-limiting kinetic step and concomitant measurements made using a chloride ion selective electrode showed that the chloride was produced quantitatively. These data will aid water utilities in predicting chloramine concentrations (and thus disinfection potential) throughout the water distribution system.


Sexual Dysfunction Among Patients With Chronic Kidney Disease.
Sexual dysfunction (SD) in patients with chronic kidney disease is common and negatively impacts quality of life. SD is often under-appreciated because of overall low awareness. Diagnosis of SD is subjective, and manifestations can be different among men and women. Causes of SD are multifactorial, including psychological disorders, hormonal imbalances, vascular disorders, neurological disorders, and medication side effects. Non-specific approaches to improving sexual function include addressing underlying psychological disorders, promoting lifestyle modifications, optimizing dialysis care, and facilitating successful kidney transplantation, whereas treatment with phosphodiesterase type 5 inhibitor, hormone replacement, and mechanical devices can be offered to patients with specific indications.


Beyond the focus group: understanding physicians' barriers to electronic medical records.
Although electronic medical records (EMRs) have potential to improve quality of care, physician adoption remains low. Rhode Island physicians' perceptions of barriers to EMRs and the association between these barriers and physician characteristics were examined. It was hypothesized that physicians with and without EMRs would differ in the types and magnitude of barriers identified. Data were drawn from the Rhode Island Department of Health's mandatory 2009 Physician Health Information Technology (HIT) survey of physicians licensed and in active practice in Rhode Island or an adjacent state. Some 1,888 (58.1% of the target population of 3,248 physicians) responded. Respondents, who were invited to provide open-ended comments, were asked to consider 11 issues as barriers to EMR use: Access to technical support, lack of computer skills, availability of a computer in the appropriate location, impact of a computer on doctor-patient interaction, lack of interoperability, privacy or security concerns, start-up financial costs, ongoing financial costs, technic limitations of systems, training and productivity impact, and lack of uniform industry standards. Respondents with EMRs consistently perceived significantly fewer barriers than those without them (p < .0001). For example, 78.9% of physicians without EMRs viewed start-up financial costs as a major barrier versus only 45.8% of physicians with EMRs. An understanding of physicians' reluctance to use EMRs is critical for developing adoption strategies. Policies to increase EMR adoption should be tailored to different physician groups to achieve maximum effectiveness. Further research into the differences between current EMR users' and nonusers' perceptions of barriers may help elucidate how to facilitate subsequent adoption.


Geriatric patient outcomes in a multidisciplinary central nervous system community hospital clinic for radiation oncology and neurosurgery (RADIANS).
Management of older adult patients with central nervous system (CNS) cancers requires a patient-centric, multidisciplinary approach. Assessment of neurosurgical and radiation treatment outcomes can assist in establishing guidelines for this patient population. We previously reported on the RADIANS clinic, a novel community hospital-based multidisciplinary clinic (MDC) for CNS cancer care, providing simultaneous radiation oncology and neurosurgery evaluation in a same-day, single-setting clinic. We now provide a focused analysis of our older adult patient population and recommendations for triage and standardization of care. Consecutive older adult patients (age ≥ 65) evaluated at the RADIANS clinic for CNS disease were identified and retrospectively reviewed. Observed 30-day neurosurgical outcomes were compared to predicted outcomes determined by the American College of Physicians NSQIP Surgical Risk Calculator. One-sample binomial exact tests were used to evaluate binary outcome measures. A two-sample t-test was used to evaluate the length of hospital stay. Brier Scores were calculated to assess the deviation between predicted probabilities and observed outcomes for binary outcome measures. Overall survival at 90 days was reported. Fifty-six older adult patients with malignant (42/56) and benign (14/56) CNS disease were evaluated. Mean distance traveled for multidisciplinary evaluation at the RADIANS clinic was 43.4 miles. There was no incidence of radiation-induced toxicity. Mean length of hospital stay for RADIANS patients was significantly shorter by about 1.5 to 3.5 days (95% CI). There was no statistically significant difference for other outcome measures, however, Brier Scores demonstrated that NSQIP was not a good predictive tool for any or serious complications, UTI, venous thromboembolism, return to OR, readmission, or death in our cohort. Local tumor control rate and progression-free survival at 90 days were 97.4% and 76.9%, respectively. This is the first report of CNS disease outcomes in older adult patients evaluated by radiation oncology and neurosurgery at a community hospital-based MDC. We observed minimal adverse radiation outcomes and high tumor control in our cohort. Findings show significantly shorter postoperative hospital stay for patients evaluated and managed at the RADIANS clinic.


Background-subtraction of fast-scan cyclic staircase voltammetry at protein-modified carbon-fiber electrodes.
Background-subtraction techniques were applied to the voltammetry of nicotinamide adenine dinucleotide (NADH) at protein-modified carbon-fiber microelectrodes. The background currents at carbon-fiber electrodes were stable and voltammetric scans immediately before or after the analyte were effectively used for background subtraction. Digital step-potential waveforms were used to excite these carbon-fiber electrodes, where the resulting voltammetric analysis assessed the optimal switching and initial potentials and the electrochemical response time was determined. The initial potential was 0.0 V and the switching potential 1.1 V (versus Ag/AgCl) and the response time was approximately 300 ms. Some sensitivity to NADH was lost and voltammetric prescans were required at protein-modified electrodes to obtain a stable baseline. Current versus time was assessed by the average current of the faradaic region from each voltammogram and by differential current; the average current minus the current from a non-faradaic potential range. Differential current assessments discriminated against artifacts caused by pH (as high as 1.0 pH unit) and ionic strength flux (100 mM). These background-subtraction techniques allowed the faradaic information to be obtained quickly and conveniently while maximizing sensitivity and maintaining selectivity.


α-synuclein promotes neuronal dysfunction and death by disrupting the binding of ankyrin to ß-spectrin.
α-synuclein plays a key role in the pathogenesis of Parkinson's disease and related disorders, but critical interacting partners and molecular mechanisms mediating neurotoxicity are incompletely understood. We show that α-synuclein binds directly to ß-spectrin. Using males and females in a Drosophila model of α-synuclein-related disorders we demonstrate that ß-spectrin is critical for α-synuclein neurotoxicity. Further, the ankyrin binding domain of ß-spectrin is required for α-synuclein binding and neurotoxicity. A key plasma membrane target of ankyrin, Na+/K+ ATPase, is mislocalized when human α-synuclein is expressed in Drosophila. Accordingly, membrane potential is depolarized in α-synuclein transgenic fly brains. We examine the same pathway in human neurons and find that Parkinson's disease patient-derived neurons with a triplication of the α-synuclein locus show disruption of the spectrin cytoskeleton, mislocalization of ankyrin and Na+/K+ ATPase, and membrane potential depolarization. Our findings define a specific molecular mechanism by which elevated levels of α-synuclein in Parkinson's disease and related α-synucleinopathies leads to neuronal dysfunction and death.


Angiotensin II type 2 receptor is expressed in human sperm cells and is involved in sperm motility.
To investigate the presence of angiotensin II type 2 receptor (AT2R) in human spermatozoa and its implication in sperm fertility status. We carried out expression assays for AT2R by reverse transcription-polymerase chain reaction, Western blot, immunofluorescence, and flow cytometry techniques in human sperm cells. Percentage of AT2R-positive sperm cells was analyzed by flow cytometry. Assisted reproduction unit and academic research laboratory. Ninety-seven human semen samples from the Clínica IVI Bilbao. All samples were examined and classified according to World Health Organization guidelines. Spermatozoa were isolated from semen on discontinuous colloidal silica gradient (45%-90%) and swim-up techniques. Presence and location of the AT2R in spermatozoa and percentage of AT2R-positive sperm cells measured by flow cytometry. We demonstrated the existence of AT2R and its transcript in human sperm by Western blot and reverse transcription-polymerase chain reaction. Immunofluorescence studies showed that AT2R is mainly located at the equatorial segment of the sperm head. The AT2R levels were associated with sperm motility parameters. Particularly, we found a significant positive correlation between AT2R and spermatozoa with progressive motility grade and a significant negative correlation with immotile spermatozoa, both in fresh semen samples and in prepared sperm cells. Regarding pathologic studies, the levels of AT2R measured by flow cytometry were lower in spermatozoa of asthenozoospermic men than in normozoospermic controls. Angiotensin II type 2 receptor is present in human semen and may be involved in the control of sperm motility. In-depth understanding of the proteins involved in sperm motility can help to elucidate the role of these proteins in male infertility as well as to establish new biomarkers for male infertility.


Resveratrol inhibits autophagy against myocardial ischemia-reperfusion injury through the DJ-1/MEKK1/JNK pathway.
Resveratrol (RES), a natural polyphenolic compound found in red wine and grape skins, has attracted significant attention due to its cardioprotective properties. DJ-1, a multifunctional protein that participated in transcription regulation and antioxidant defense, was shown to provide a significant protective impact in cardiac cells treated with ischemia-reperfusion. We created a myocardial ischemia-reperfusion (I/R) model in vivo and in vitro by ligating the left anterior descending branch of rats and subjecting H9c2 cells to anoxia/reoxygenation (A/R) to investigate whether RES reduces myocardial ischemia-reperfusion injury by upregulating DJ-1. We discovered that RES dramatically enhanced cardiac function in rats with I/R. Subsequently, we found that RES prevented the rise in autophagy (P62 degradation and LC3-II/LC3-I increase) induced by cardiac ischemia-reperfusion in vitro and in vivo. Notably, the autophagic agonist rapamycin (RAPA) eliminated RES-induced cardioprotective effects. In addition, Further data showed that RES significantly increased the expression of DJ-1 in the myocardium with the treatment of I/R. At the same time, pretreatment with RES reduced phosphorylation of MAPK/ERK kinase kinase 1 (MEKK1) and Jun N-terminal Kinase (JNK) stimulated by cardiac ischemia-reperfusion, and Beclin-1 mRNA and protein levels while decreasing lactate dehydrogenase (LDH) and improving cell viability. However, the lentiviral shDJ-1 and JNK agonist anisomycin disrupted the effects of RES. In summary, RES could inhibit autophagy against myocardial ischemia-reperfusion injury through DJ-1 modulation of the MEKK1/JNK pathway, providing a novel therapeutic strategy for cardiac homeostasis.


Breast cancer diagnosis by scintimammography: a meta-analysis and review of the literature.
Scintimammography is a relatively new, non-invasive diagnostic modality in the evaluation of breast cancer. The purpose of the current study was to review the existing literature on the accuracy of scintimammography in the diagnosis of breast cancer. A search of all articles published between 1st January 1967 and 31st December 1999 was conducted. A total of 64 unique studies were selected. Each scientific paper was reviewed for scientific merit by an epidemiologist, a surgeon and a surgical resident. Assessment of scientific merit was based on a scoring scheme developed for the study. The articles included in this review reported data on a total of 5340 patients assessed for breast cancer with scintimammography. The aggregated summary estimates on these patients were sensitivity: 85.2% and specificity: 86.6%. For patients with a palpable mass the sensitivity and specificity were 87.8 and 87.5%, respectively. For patients without a palpable mass the sensitivity was 66.8% and that for specificity was 86.9%. The results of this review have shown that scintimammography may be used effectively as an adjunct to mammography and physical examination in the diagnosis of breast cancer.


Presence of an abnormal beta 2-microglobulin mRNA in Daudi cells: induction by interferon.
Human alpha and beta interferons increase the amount of class I human histocompatibility messenger RNA HLA-A, B, C and beta 2-microglobulin in most human cells studied to date. This report concerns the effect of interferons on the Burkitt lymphoma-derived cell line Daudi, which does not express HLA-A, B, C antigens or beta 2-microglobulin on its membrane. HLA-A, B, C messenger RNA present in Daudi cells is increased by both alpha and beta interferons. Furthermore, we have shown that although it was not possible to detect mature beta 2-microglobulin protein in the cytoplasm or on the cell membrane of Daudi cells, a poly A+ messenger RNA is present in Daudi cells, which hybridizes with a cDNA clone specific for human beta 2-microglobulin. This abnormal messenger RNA is, however, increased normally by interferon. These effects were also observed with human interferon beta on a variant of Daudi cells characterized by a markedly reduced sensitivity to anti-proliferative and anti-cellular effects of human interferon alpha.


'I was terrified for my child': understanding the link between the Dengvaxia® controversy and the measles vaccine hesitancy in Pasay City, Philippines.
The Dengvaxia® (dengue vaccine) controversy has been identified as one of the main reasons for the measles vaccine hesitancy in the Philippines. Our study aimed to identify various issues related to the Dengvaxia® controversy and to link these issues with the social perspective of measles vaccine refusal. Semi-structured interviews and a focus group discussion using ethnography research were conducted with 41 parents and healthcare workers in Pasay City. Using Victor Turner's Social Drama Theory, our study identified existing social issues relating to the different angles of the Dengvaxia® controversy and the measles vaccine hesitancy. Misinformation on the failed Dengvaxia® rollout implementation has challenged the fundamental understanding of the importance of immunization programs. Our findings on vaccine hesitancy in the community showed a complex problem with compounded factors, including medical populism, moral panics and other social views. We described how Pasay City's clinic waiting room became a significantly important scenario where individuals often discuss information, concerns and experiences on vaccines and vaccine hesitancy. Our study suggests that the Dengvaxia® controversy may reduce the measles vaccination confidence in the Philippines. Lack of transparency played a crucial role in this dilemma, producing a cascading effect on the other vaccines' safety.


Expressive suppression and neural responsiveness to nonverbal affective cues.
Optimal social functioning occasionally requires concealment of one's emotions in order to meet one's immediate goals and environmental demands. However, because emotions serve an important communicative function, their habitual suppression disrupts the flow of social exchanges and, thus, incurs significant interpersonal costs. Evidence is accruing that the disruption in social interactions, linked to habitual expressive suppression use, stems not only from intrapersonal, but also from interpersonal causes, since the suppressors' restricted affective displays reportedly inhibit their interlocutors' emotionally expressive behaviors. However, expressive suppression use is not known to lead to clinically significant social impairments. One explanation may be that over the lifespan, individuals who habitually suppress their emotions come to compensate for their interlocutors' restrained expressive behaviors by developing an increased sensitivity to nonverbal affective cues. To probe this issue, the present study used functional magnetic resonance imaging (fMRI) to scan healthy older women while they viewed silent videos of a male social target displaying nonverbal emotional behavior, together with a brief verbal description of the accompanying context, and then judged the target's affect. As predicted, perceivers who reported greater habitual use of expressive suppression showed increased neural processing of nonverbal affective cues. This effect appeared to be coordinated in a top-down manner via cognitive control. Greater neural processing of nonverbal cues among perceivers who habitually suppress their emotions was linked to increased ventral striatum activity, suggestive of increased reward value/personal relevance ascribed to emotionally expressive nonverbal behaviors. These findings thus provide neural evidence broadly consistent with the hypothesized link between habitual use of expressive suppression and compensatory development of increased responsiveness to nonverbal affective cues, while also suggesting one explanation for the suppressors' poorer cognitive performance in social situations. Moreover, our results point to a potential neural mechanism supporting the development and perpetuation of expressive suppression as an emotion regulation strategy.


Quantification of IgG anti-D bound to D-positive red cells infused into D-negative subjects after intramuscular injection of monoclonal anti-D.
A flow-cytometric method was developed to determine the number of molecules of IgG bound to D-positive red blood cells (RBC) when sensitized with low plasma concentrations of IgG anti-D in the presence of an excess of D-negative RBC. D-positive RBC were infused into 12 D-negative male volunteers 2 days after intramuscular injection of monoclonal anti-D (BRAD-3, IgG3 or BRAD-5, IgG1). Blood samples were taken immediately before, 3 min and 3 h after injection of the RBC, incubated for 1 h at 37 degrees C, washed, then incubated sequentially with FITC-conjugated anti-IgG, IgM monoclonal anti-D, biotin-conjugated anti-IgM, and R-phycoerythrin-conjugated streptavidin. To prepare a standard curve, O R1R2 RBC were incubated in duplicate with varying dilutions of BRAD-3 or BRAD-5, and RBC from one set were mixed with an excess of D-negative RBC (1:100) and labelled as above, while cell-bound IgG on the other set was quantified by ELISA. The test samples and standards were analyzed by flow cytometry and the mean channel (FITC) fluorescence was plotted against molecules IgG/cell, from which the sensitization level of D-positive RBC in the test samples was determined. The use of IgM anti-D enhanced the discrimination between D-positive and D-negative RBC, especially when fewer than about 3000 molecules IgG/cell were bound. The assay was sensitive to about 1000 molecules IgG/cell. The sensitization levels of the D-positive RBC in samples taken 3 h after injection were found to be in the range 1500-10,000 molecules IgG anti-D per cell.


Bronchoprotection with formoterol via dry powder and metered-dose inhalers in patients with asthma.
Determination of device comparability for new inhaled medications is essential. To evaluate the methacholine bronchoprotection afforded by formoterol, 12 microg, delivered via Clickhaler and Aerolizer dry powder inhalers and a pressurized metered-dose inhaler (pMDI) in mild-to-moderate asthmatic patients. Two separate randomized, double-blind, double-dummy, crossover studies were performed. Peak bronchoprotection (30 minutes) was measured after administration of a single dose in a placebo-controlled study (n = 16). First-dose (8 hours) and trough (12 hours) protection were evaluated after 2 weeks (n = 28). Doubling dilution differences (DDD) in the methacholine provocation concentration that caused a decrease in forced expiratory volume in 1 second of 20% from baseline were compared, with equivalence defined if the 95% confidence interval was within the predefined equivalence limits of +/- 1.5 DDD (peak) and +/- 1.0 DDD (trough). For peak single-dose effects, DDD (95% confidence interval) data showed significant protection vs placebo for all devices but no significant differences among active inhalers. For trough first-dose effects, there was significant protection vs baseline for all devices and equivalence for Clickhaler vs Aerolizer (-0.41; -0.85 to 0.04) and Aerolizer vs pMDI (-0.27; -0.66 to 0.13) but not for Clickhaler vs pMDI (-0.68; -1.12 to -0.23). For trough effects after 2 weeks, there was significant residual protection from baseline and equivalence for Clickhaler vs Aerolizer (-0.32; -0.94 to 0.30), Clickhaler vs pMDI (0.34; -0.96 to 0.27), and Aerolizer vs pMDI (-0.02; -0.57 to 0.52). Formoterol delivered by 3 different inhalers exhibited a significant degree of peak and trough bronchoprotection after single and repeated dosing, with most comparisons being within predefined equivalence limits.


Polycyclic aromatic hydrocarbons in soils along the coastal and estuarine areas of the northern Bohai and Yellow Seas, China.
Polycyclic aromatic hydrocarbons (PAHs) and their health risks in surface soils (n = 31) collected from coastal and estuarine areas of the northern Bohai and Yellow Seas (CEANBYS), China, were investigated. Total concentrations of PAHs ranged from 6.6 × 10(1) to 9.2 × 10(2) ng g(-1) dry weight, with an average of 3.1 × 10(2) ng g(-1) dw. The locations where greater concentrations of PAHs were observed were all near factories emitting black smoke or on the edge of the urban areas. These observations are consistent with concentrations of PAHs in soils being influenced by human activities, especially industrialization and urbanization. Concentrations of PAHs were significantly correlated with concentrations of organic carbon in soils. The patterns of relative concentrations and types of PAHs observed as well as knowledge of the potential sources were consistent with the primary sources of PAHs in soils of the CEANBYS being derived from the pyrolytic processes such as combustion of fossil fuel. The incremental lifetime cancer risks of exposing to PAHs for child, youth, and adult were 1.6 × 10(-6), 1.2 × 10(-6), and 1.9 × 10(-6).


Drug dealers: $20 trillion of in-licensing payments.
To help understand the trends of in-licensing, we analysed 781 in-licensing deals costing over $20 trillion over a period of 17 years and found that, while the number of FDA approvals is decreasing, the upfront price paid for drug in-licensing is increasing. Moreover, these upfront payments were not immune to the recent economic downturn, where the yearly increase in upfront payments halted in years 2008-2012. The cost of buying drugs also steadily increased from early research compounds to Phase III drugs. A relative decrease in upfront payments in Filed phase versus to Phase III was seen, which may reflect reluctance in buying filed drugs that are facing the 'patent cliff'. The data here also showed no obvious difference in upfront payments made in five major disease areas, namely, oncology, cardiovascular disease, autoimmunity, nervous system and metabolic disorders.


Lidocaine for status epilepticus in adults.
Our goal was to perform a systematic review of the literature on the use of intravenous lidocaine in adults for status epilepticus (SE) and refractory status epilepticus (RSE) to determine its impact on seizure control. All articles from MEDLINE, BIOSIS, EMBASE, Global Health, HealthStar, Scopus, Cochrane Library, the International Clinical Trials Registry Platform (inception to November 2014), and gray literature were searched. The strength of evidence was adjudicated using both the Oxford and GRADE methodology by two independent reviewers. Overall, 13 studies were identified, with 11 manuscripts and 2 meeting abstracts. Seventy-six adult patients were treated for 82 episodes of SE/RSE. Patients had varying numbers of anti-epileptic drugs (AEDs), 1-12, on board prior to lidocaine therapy. During 69 of the 82 (84.1%) episodes of SE/RSE, phenytoin was on board. The dose regimen of lidocaine varied, with some utilizing bolus dosing alone; others utilizing a combination of bolus and infusion therapy. Overall, 70.7% of seizures responded to lidocaine, with complete cessation and greater than 50% reduction seen in 64.1% and 6.1% respectively. Patient outcomes were sparingly reported. There currently exists level 4, GRADE C evidence to support the consideration of lidocaine for SE and RSE in the adult population. Thus there is currently weak evidence to support the use of lidocaine in this context. Further prospective studies of lidocaine administration in this setting are warranted.


Enlarging arachnoid cyst: a false alarm for infants.
The natural history of change in size of arachnoid cysts (ACs) is an unsettled issue. This leads to confusion in deciding treatment strategies of incidental, asymptomatic AC in children, since the risks and complications of surgery are not negligible. The aim of the present study was to elucidate the clinical significance of change in size of AC in young children. From 2000 to 2008, 86 patients younger than 5 years with AC documented on brain imaging were included. They were divided into four different age groups (group I: 0-0.5 year, group II: 0.5-1 year, group III: 1-3 years, group IV: 3-5 years). The temporal changes in size of the AC on consecutive ultrasonography, computed tomography (CT), or magnetic resonance imaging (MRI) of each patient were measured and plotted as individual graphs. Natural progression of AC size was assessed by evaluation of the pattern of each graph. Seventeen patients showed enlargement of AC during follow-up; 14 patients were younger than 1 year. None of the patients in group IV showed enlargement during follow-up. Overall, 11 of the 17 patients with demonstrated AC enlargement were not operated on. In six patients, AC enlargement ceased, and spontaneous regression was observed in three patients. The remaining two untreated patients had shown asymptomatic increase in size at last follow-up. Statistical analysis revealed age at diagnosis to be the only significant factor affecting cyst enlargement. Asymptomatic enlargement of AC in infants may not be an indication for surgical intervention. Close follow-up is a viable option for such children.


Tolerability of beta-blockers in elderly patients with chronic heart failure: the COLA II study.
Beta-blockers are recommended therapy for patients with chronic heart failure (CHF). However, there remains concern regarding tolerability of these agents in the elderly, which has contributed to the limited uptake of these agents in clinical practice. We conducted a multi-national, prospective evaluation of tolerability to carvedilol in 1030 CHF patients aged >70 years selected by their treating physician to receive this agent in everyday practice. NYHA Class II-IV CHF patients were assessed at baseline for key demographic parameters that may predict tolerability, then followed for 6 months after starting carvedilol. Tolerability was defined as being on >or=6.25 mg bd of carvedilol at 6 months having received a total of >or=3 months therapy. Tolerability overall was 80% with age 70-75 years 84.3%, 76-80 years 76.8% and >80 years 76.8%. Mean carvedilol dose achieved was 31.2 mg. In multivariate analysis, advanced age, low diastolic BP, LVEF, obstructive airways disease and presence of diabetes were predictors of tolerability. Carvedilol appears to be well tolerated in this elderly CHF patient cohort. Therefore, elderly CHF patients should not be denied treatment with carvedilol because of concerns regarding tolerability.


Increased cortisol excretion in chronic pain.
Urine samples were collected from 67 patients attending a Regional Pain Unit for treatment of chronic pain. Patients treated with steroids or with any signs or symptoms of Cushing's syndrome were excluded. The urine cortisol creatinine ratio was subsequently measured. Values for the urine cortisol/creatinine ratio were elevated in 31% of the patients, and grossly elevated in some. These patients could not be differentiated from those with normal ratios on the basis of pain or mood scores, cause of pain or drug therapy; they tended to have pain of shorter duration. Possible interpretations of these results are discussed.


Bursts of beta oscillation differentiate postperformance activity in the striatum and motor cortex of monkeys performing movement tasks.
Studies of neural oscillations in the beta band (13-30 Hz) have demonstrated modulations in beta-band power associated with sensory and motor events on time scales of 1 s or more, and have shown that these are exaggerated in Parkinson's disease. However, even early reports of beta activity noted extremely fleeting episodes of beta-band oscillation lasting <150 ms. Because the interpretation of possible functions for beta-band oscillations depends strongly on the time scale over which they occur, and because of these oscillations' potential importance in Parkinson's disease and related disorders, we analyzed in detail the distributions of duration and power for beta-band activity in a large dataset recorded in the striatum and motor-premotor cortex of macaque monkeys performing reaching tasks. Both regions exhibited typical beta-band suppression during movement and postmovement rebounds of up to 3 s as viewed in data averaged across trials, but single-trial analysis showed that most beta oscillations occurred in brief bursts, commonly 90-115 ms long. In the motor cortex, the burst probabilities peaked following the last movement, but in the striatum, the burst probabilities peaked at task end, after reward, and continued through the postperformance period. Thus, what appear to be extended periods of postperformance beta-band synchronization reflect primarily the modulated densities of short bursts of synchrony occurring in region-specific and task-time-specific patterns. We suggest that these short-time-scale events likely underlie the functions of most beta-band activity, so that prolongation of these beta episodes, as observed in Parkinson's disease, could produce deleterious network-level signaling.


Is inflammation a significant predictor of bile duct injury during laparoscopic cholecystectomy?
Bile duct injuries (BDI) have been reported to occur more frequently during laparoscopic cholecystectomy (LC) compared to open cholecystectomy (OC). Several studies have demonstrated various potential predisposing factors for BDI. However, there is a controversy as to whether gallbladder inflammation is a significant predictor for BDI. Therefore, out primary aim was to investigate the relationship between inflammation and BDI at LC, and secondarily to present the management and clinical outcome of BDI. We recorded all consecutive LC performed between 1993 and 2005 in our institution by nine staff surgeons. BDI were classified according to Strasberg's classification. Simple and multivariate logistic regression analysis was performed to evaluate the association between inflammation and BDI occurrence during LC. There were 2,184 patients. Among those, 344 had inflammation (16%). The conversion rate was 5% and was higher among male, elder patients, and those with inflammation. The BDI incidence was 0.69% (0.14% for major and 0.55% for minor injuries) and it was significantly higher in those with inflammation compared to those without inflammation (p = 0.01). In particular, the risk for BDI was almost 3.5 times higher in those with inflammation (OR = 3.61, 95% CI 1.27-10.21). Inflammation remained an independent risk factor for BDI even after adjustment for potential confounders. Among patients sustaining injury, one died and two have recurrent cholangitis. No association was observed between clinical outcome and management of BDI, time of diagnosis, sex, and inflammation. We revealed that inflammation is an independent predictor of BDI occurrence during LC. Therefore, it would be advisable for surgeons to not hesitate to convert a LC to an OC in the presence of inflammation.


Alternative promoter usage of the Fos-responsive gene Fit-1 generates mRNA isoforms coding for either secreted or membrane-bound proteins related to the IL-1 receptor.
Fit-1 has been identified previously as a Fos-responsive gene of rat fibroblasts. Here we show that Fit-1 is directly regulated by the estrogen-inducible transcription factor Fos-ER and that it belongs to the family of delayed early genes. Two different mRNA isoforms are expressed from the Fit-1 gene. The Fit-1M mRNA isolated from spleen codes for a membrane-bound protein which is most closely related in its extracellular, transmembrane and intracellular domains to the type I interleukin-1 (IL-1) receptor. The Fit-1S mRNA of fibroblasts directs, instead, the synthesis of a secreted protein consisting of only the extracellular domain. Analysis of the exon-intron structure of the Fit-1 gene indicated that the Fit-1S and Fit-1M mRNAs are transcribed from two different promoters and that the sequence differences at their 3' ends result from alternative 3' processing. Northern blot analysis with specific 5' and 3' probes directly demonstrated tight coupling between alternative promoter usage and 3' processing of the Fit-1 transcripts. The orthologous gene of the mouse (known as T1 or ST2) is expressed during ontogeny first in the fetal liver of the embryo and then in lung and hematopoietic tissues of the adult. The mRNA coding for the membrane-bound protein is more abundantly expressed in all of these tissues, while the transcript for the secreted form predominates in fibroblasts and mammary epithelial cells. Differential regulation of two distinct promoters is thus used to determine the ratio between secreted and membrane-bound forms of Fit-1 (T1/ST2) which may modulate signaling in response to IL-1.


Triple-hurdle model analysis of the factors influencing biogas digester building, use and processing by Chinese pig farmers.
Because of the pollution associated with pig manure, pig farmers are being encouraged to adopt environmentally friendly manure management solutions such as biogas digesters. However, as the correlations and influencing factors associated with the different manure disposal stages remain unclear, these solutions have failed to positively change farmer disposal behaviors. Therefore, this paper constructed a triple-hurdle model to empirically analyze the pig farmer manure disposal behaviors and the associated relationships under the belief that the three stage biogas digester decisions were driven by structurally different processes. This paper adds to the literature in three ways: (1) it provides a dynamic framework that identifies pig farmer manure disposal behaviors; (2) it applies a recently developed econometric method that corrects for conditional correlations between error terms; and (3) proves that certain factors might have opposite impacts in different behavior stages. It was found that cultivated land area and farm income ratio had statistically significant positive impacts on biogas digesters construction behavior but a negative influence on the farmers' use of the biogas digesters. Farmers who joined a cooperative were found to be statistically more likely to be biogas digester users, and the unconditional expected share of the actual treated manure was significantly higher. This study can assist policymakers in formulating and implementing strategies to encourage greater biogas digestor use by pig farmers.


Production and processing of antioxidant bioactive peptides: A driving force for the functional food market.
Recently, the demand for functional foods in the global market has increased rapidly due to the increasing occurrences of non-communicable diseases and technological advancement. Antioxidant peptides have been suggested as ingredients used to produce health-promoting foods. These peptides are encrypted from various food derived protein sources by chemical and enzymatic hydrolysis, and microbial fermentation. However, the industrial-scale production of antioxidant peptides is hampered by different problems such as high production cost, and low yield and bioactivity. Accordingly, novel processing technologies, such as high pressure, microwave and pulsed electric field, have been recently emerged to overcome the problems associated with the conventional hydrolysis methods. This particular review, therefore, discussed the current processing technologies used to produce antioxidant peptides. The review also suggested further perspectives that should be addressed in the future.


Arthrobacter soli sp. nov., a novel bacterium isolated from wastewater reservoir sediment.
A novel Gram-positive bacterium, designated SYB2T, was isolated from wastewater reservoir sediment, and a polyphasic taxonomic study was conducted based on its morphological, physiological, and biochemical features, as well as the analysis of its 16S rRNA gene sequence. During the phylogenetic analysis of the strain SYB2T, results of a 16S rRNA gene sequence analysis placed this bacterium in the genus Arthrobacter within the family Micrococcaceae. SYB2T and Arthrobacter protophormiae ATCC 19271T, the most closely related species, both exhibited a 16S rRNA gene sequence similarity of 98.99%. The genomic DNA G+C content of the novel strain was found to be 62.0 mol%. The predominant fatty acid composition was anteiso-C15:0, anteiso-C17:0, iso-C16:0, and iso-C15:0. Analysis of 16S rRNA gene sequences and DNA-DNA relatedness, as well as physiological and biochemical tests, showed genotypic and phenotypic differences between strain SYB2T and other Arthrobacter species. The type strain of the novel species was identified as SYB2T (= KCTC 19291T= DSM 19449T).


Extracting drug-enzyme relation from literature as evidence for drug drug interaction.
Information about drug-drug interactions (DDIs) is crucial for computational applications such as pharmacovigilance and drug repurposing. However, existing sources of DDIs have the problems of low coverage, low accuracy and low agreement. One common type of DDIs is related to the mechanism of drug metabolism: a DDI relation may be caused by different interactions (e.g., substrate, inhibit) between drugs and enzymes in the drug metabolism process. Thus, information from drug enzyme interactions (DEIs) serves as important supportive evidence for DDIs. Further, potential DDIs present implicitly could be detected by inference and reasoning based on DEIs. In this article, we propose a hybrid approach to combining machine learning algorithm with trigger words and syntactic patterns, for DEI relation extraction from biomedical literature. The extracted DEI relations are used for reasoning to infer potential DDI relations, based on a defined drug-enzyme ontology incorporating biological knowledge. Evaluation results demonstrate that the performance of DEI relation extraction is promising, with an F-measure of 84.97% on the in vivo dataset and 65.58% on the in vitro dataset. Further, the inferred DDIs achieved a precision of 83.19% on the in vivo dataset and 70.94% on the in vitro dataset, respectively. A further examination showed that the overlaps between our inferred DDIs and those present in DrugBank were 42.02% on the in vivo dataset and 19.23 % on the in vitro dataset, respectively. This paper proposed an effective approach to extract DEI relations from biomedical literature. Potential DDIs not present in existing knowledge bases were then inferred based on the extracted DEIs, demonstrating the capability of the proposed approach to detect DDIs with scientific evidence for pharmacovigilance and drug repurposing applications.


Increased Body Mass Index Augments Endothelial Injury and Clinical Outcomes after Hematopoietic Stem Cell Transplantation.
Higher body mass index (BMI) is characterized as a chronic inflammatory state with endothelial dysfunction. Endothelial injury after allogeneic hematopoietic stem cell transplantation (allo-HSCT) puts patients at risk for such complications as transplantation-associated thrombotic microangiopathy (TA-TMA) and acute graft-versus-host-disease (aGVHD). To evaluate the impact of increased BMI on endothelial injury after allo-HSCT in pediatric and young adult patients, we conducted a retrospective cohort study evaluating 476 consecutive allo-HSCT children and young adult recipients age 0 to 20 years. Our analysis was subdivided based on distinct age categories (<2 years and 2 to 20 years). BMI was considered as a variable but was also expressed in standard deviations from the mean adjusted for age and sex (z-score), based on established criteria from the World Health Organization (age <2 years) and the Centers for Disease Control and Prevention (age 2 to 20 years) to account for differences associated with age. Primary endpoints included the incidences of TA-TMA and aGVHD. Increased BMI z-score was associated with TA-TMA after allo-HSCT in patients age <2 years (median, 18.1; IQR, 17 to 20; P = .006) and in patients age 2 to 20 years (median, 18.7; IQR, 16 to 21.9; P = .02). Higher BMI z-score correlated with TA-TMA risk in both age groups, with a BMI z-score of .9 in the younger cohort and .7 (IQR, -.4 to 1.6; P = .04) in the older cohort. Increased BMI z-score was associated with an increased risk of TA-TMA in a multivariate analysis of the entire cohort (odds ratio [OR], 1.2; 95% confidence interval [CI], 1.05 to 1.37; P = .008). Multivariate analysis also demonstrated that patients with BMI in the 85th percentile or greater had an increased risk of developing TA-TMA compared to those with a lower BMI percentile (OR, 2.66; 95% CI, 1.62 to 4.32; P < .001). Baseline and day +7 ST2 levels were elevated in subjects with TA-TMA compared to those without TA-TMA in both age groups. Baseline sC5b-9 concentration was not correlated with BMI z-score, but sC5b-9 concentration was increased markedly by 7 days post-allo-HSCT in patients age <2 years who later developed TA-TMA compared to those who never developed TA-TMA (P = .001). The median BMI z-score was higher for patients with aGVHD compared to patients without aGVHD (.7 [range, -3.9 to 3.9] versus .2 [range, -7.8 to 5.4]; P = .03). We show that high BMI is associated with augmented risk of endothelial injury after HSCT, specifically TA-TMA. These data identify a high-risk population likely to benefit from early interventions to prevent endothelial injury and prompt treatment of established endothelial injury.


Interprofessional communication in a psychiatric outpatient unit - an ethnographic study.
Communication in healthcare has been extensively studied, but most research has focused on miscommunication and the importance of communication for patient safety. Previous research on interprofessional communication has mainly focused on relationships between physicians and nurses in non-psychiatric settings. Since communication is one of the core competencies in psychiatric care, more research on interprofessional communication between other clinicians is needed, and should be explored from a broader perspective. This study aimed to explore and describe interprofessional communication in a psychiatric outpatient unit. During spring 2022, data consisting of over 100 h of fieldwork were collected from observations, formal semi-structured interviews and informal conversations inspired by the focused ethnography method. Data was collected at an outpatient unit in central Sweden, and various clinicians participated in the study. The data analysis was a back-and-forth process between initial codes and emerging themes, but also cyclical as the data analysis process was ongoing and repeated and took place simultaneously with the data collection. We found that a workplace's history, clinicians´ workload, responsibilities and hierarchies influence interprofessional communication. The results showed that the prerequisites for interprofessional communication were created through the unit's code of conduct, clear and engaging leadership, and trust in the ability of the various clinicians to perform new tasks. Our results indicate that leadership, an involving working style, and an environment where speaking up is encouraged and valued can foster interprofessional communication and respect for each other´s professional roles is key to achieving this. Interprofessional communication between different clinicians is an important part of psychiatric outpatient work, where efficiency, insufficient staffing and long patient queues are commonplace. Research can help shed light on these parts by highlighting aspects influencing communication.


Longitudinal analyses suggest a higher burden of self-reported influenza-like illness among women and girls in a rural community in western Kenya.
Longitudinal studies that quantify influenza-like illness (ILI) burden while determining associated risk factors and intra-cluster correlation coefficients (ICCs) are scarce in Africa. This study aimed to identify longitudinal patterns of ILI and the associated independent variables while quantifying the burden by age and sex. We enrolled and followed 390 subjects nested within 92 households for 12 weeks, collecting weekly data on prespecified self-reported symptoms in a rural community in western Kenya. Using a standard case definition, reporting at least any four prespecified symptoms attained an ILI syndrome. Analyses related relationship between attaining ILI syndrome and independent variables using multilevel logistic regression models adjusting for age and sex. ICCs quantified clustering of ILI syndrome computed at different levels of data aggregation. Of the 4261 reports received, 592 (14%) registered an illness, with 278 (48%) of these attaining ILI syndrome. The 278 reports belonged to 176 (41%) subjects dwelling in 76 (83%) households. On average, each study subject and household experienced 1.6 and 3.7 episodes of ILI syndrome, respectively. Making a visit outside the local area of residence (Odds ratio (OR) =2.7, 95% CI 1.8, 4.1) and living in a cement-floored house (OR = 1.9, 95% CI 1.1, 3.3) independently predicted attaining ILI syndrome. Correcting for the population structure revealed an excess risk of 16% in attaining the syndrome among the study women and girls. Clustering of ILI syndrome was lowest at the individual level (ICC = 0.04) and slightly higher within households (ICC = 0.14), suggesting moderate household contextual influences. Regardless of the cause, we provide evidence of a potentially higher ILI syndrome burden among women and girls in our study area. Studies are needed to establish granular exposures defining the increasing risk of ILI syndrome associated with gender, making a visit outside the local sub-county of residence and living in a cement-floored house.


Effectiveness of Non-pharmacologic Interventions on Device-measured Physical Activity in Adults With Cancer, and Methodology Used for Assessment: A Systematic Review and Meta-analysis.
To investigate the effectiveness of different types of interventions aimed at enhancing device-measured physical activity (PA) and summarize the devices and methodologies used to measure PA in adults with cancer. A systematic review was prospectively registered on PROSPERO (CRD42020199466). The search was conducted in PubMed, The Cochrane Library, EMBASE (via Ovid), and PEDro from 2005 onward. Prospective interventional studies (randomized controlled trials [RCTs], non-randomized controlled trials, and single-group trials), that included adults within 12 months from cancer diagnosis, and device-measured PA before and after commencement of an intervention were included. Studies were excluded if PA was measured at a single time point. Two independent reviewers screened 3033 records and 30 articles met the inclusion criteria. Two reviewers independently extracted the data. PEDro scale and GRADE approach were used to assess methodological quality of RCTs and overall quality of evidence, respectively. A meta-analysis of relevant RCTs was conducted. Thirty studies were identified, mainly including adults with multiple cancer types. Interventions were behavior change interventions (n=15), exercise training (n=13), neuromuscular electrostimulation (n=1), or a nutritional program (n=1). The meta-analysis showed improvements on moderate-to-vigorous intensity PA (MVPA) in the experimental group (8 studies; standardized mean difference (SMD)=0.23; 95% CI 0.06-0.39); with subgroup analysis showing that findings were mainly driven by behavior change interventions (5 studies; SMD=0.23, 95% CI 0.05-0.41). An uncertain effect on sedentary behavior, daily steps, and light intensity PA was found. PA was measured with medical devices and commercial wearables, quality of the methodology was variable. Behavior change interventions increased device-derived MVPA in adult cancer patients who underwent the intervention within 12 months of the cancer diagnosis. Various devices and methodologies were used to assess PA, which limits comparisons across the studies.


Initial experience with Symbios Model 7008 pacemaker.
The Symbios 7008 pacemaker is capable of treating both bradycardias and tachycardias. In this report, we summarize operation of the pacemaker and the initial experience in 21 patients who have received implants. All patients have supraventricular tachyarrhythmias that have been terminated successfully with automatic atrial burst pacing (19 patients) or AV sequential pacing. More than half of the patients also have bradyarrhythmias requiring pacing and thus benefit from both functions of the device. Experience to date has been favorable.


Incidence of sudden cardiac arrest and sudden cardiac death after unstable angina pectoris and myocardial infarction.
Sudden cardiac arrests (SCA) and sudden cardiac deaths (SCD) are believed to account for a large proportion of deaths due to cardiovascular causes. The purpose of this study is to provide comprehensive information on the epidemiology of SCAs and SCDs after acute coronary syndrome. The incidence of SCA (including SCDs) was studied retrospectively among 10,316 consecutive patients undergoing invasive evaluation for acute coronary syndrome (ACS) between 2007 and 2018 at Tays Heart Hospital (sole provider of specialized cardiac care for a catchment area of over 0.5 million residents). Baseline and follow-up information was collected by combining information from the hospital's electronic health records, death certificate data, and a full-disclosure review of written patient records and accounts of the circumstances leading to death. During 12 years of follow-up, the cumulative incidence of SCAs (including SCDs) was 9.8% (0.8% annually) and that of SCDs 5.4% (0.5% annually). Cumulative incidence of SCAs in patients with ST-elevation myocardial infarction, non-ST-elevation myocardial infarction and unstable angina pectoris were: 11.9%,10.2% and 5.7% at 12 years. SCAs accounted for 30.5% (n = 528/1,732) of all deaths due to cardiovascular causes. The vast majority of SCAs (95.6%) occurred in patients without implantable cardioverter defibrillator (ICD) devices or among patients with no recurrent hospitalizations for coronary artery disease (89.1%). SCAs accounted for less than a third of all deaths due to cardiovascular causes among patients with previous ACS. Incidence of SCA is highest among STEMI and NSTEMI patients. After the hospital discharge, most of SCAs happen to NSTEMI patients.


A scoping review of methods used in musculoskeletal soft tissue and nerve shear wave elastography studies.
This scoping review of shear wave elastography (SWE) articles in musculoskeletal soft tissue and nerve research demonstrates methodological heterogeneity resulting from a lack of standardized data collection and reporting requirements. Seven literature databases were searched for original articles published in English from 2004-2020 that examine human skeletal muscles, tendons, and nerves in vivo. Although 5,868 records were initially identified, only 375 reports met inclusion criteria. Of the 375 articles, 260 examined 89 unique muscles, 94 examined 14 unique tendons, and 43 examined 8 unique nerves. Cohorts were often small (n = 11-20) and young (mean = 20-29 years), and participants were typically tested in the prone position. Regarding equipment, a variety of ultrasound systems (n = 11), ultrasound models (n = 18), and transducers (n = 19) were identified. Only 11% of articles contained information on the use of electromyography to confirm absence of muscle activity, and only 8% reported measurement depth. Since musculoskeletal soft tissue and nerve stiffness can vary significantly based on data collection methods, it is essential to standardize SWE collection and reporting procedures. This will allow SWE to serve as a valid and reproducible tool for assessing tissue pathology, disease progression, and response to intervention within a variety of musculoskeletal and nerve-related disorders.


Pharmacokinetics and Pharmacodynamics of NPH Insulin in Type 1 Diabetes: The Importance of Appropriate Resuspension Before Subcutaneous Injection.
Crystalline NPH insulin comes in a two-phase solution with either a solvent or a rapid-acting insulin (in premixed formulations) and needs adequate mixing for complete resuspension before injection. The aim of this study was to establish pharmacokinetics (PK) and pharmacodynamics (PD) after injection of appropriately resuspended versus nonresuspended NPH insulin. PK and PD were assessed after subcutaneous injection of NPH insulin 0.35 units/kg at steady state by pen either resuspended (R+, tipping of insulin pen 20 times) or nonresuspended (pen maintained in fixed position either horizontally [R- horizontal] or vertically with tip up [R- up] or tip down [R- down]). Eleven subjects with type 1 diabetes (age 31.5 ± 12 years, diabetes duration 17.5 ± 7.7 years, BMI 22.9 ± 1.5 kg/m2, A1C 7.2 ± 0.4% [55.2 ± 4.4 mmol/mol]) were studied (euglycemic clamp) with a randomized crossover design. Compared with resuspended NPH insulin (R+), nonresuspended NPH insulin resulted in profound PK/PD differences with either reduced (R- horizontal and R- up) or increased (R- down) plasma insulin concentrations [FIRI_AUC(0-end of study) (free immunoreactive insulin area under the concentration-time curve between 0 and end of study)] and PD activity [glucose infusion rate (GIR)_AUC(0-end of study)] (all P < 0.05). Duration of NPH insulin action was shorter in R- up (9.4 ± 1.7 h) but longer in R- down (15.4 ± 2.3 h) compared with R+ (11.8 ± 2.6 h) (P < 0.05). Within-subject variability (percent coefficient of variation) among studies was as high as 23% for PK [FIRI_AUC(0-end of study)] and 62% for PD [GIR_AUC(0-end of study)]. Compared with resuspended NPH insulin, lack of resuspension profoundly alters PK/PD and may importantly contribute to day-to-day glycemic variability of type 1 diabetes.


HIV therapies and the kidney: some good, some not so good?
Several observational studies have identified tenofovir as an independent risk factor for kidney impairment. Conversely, randomized trials have only demonstrated minor tenofovir-related changes in kidney function, but these studies included patients with normal kidney function and with low underling risk for progression of their renal function, had limited size, and limited follow-up. Several potential mechanisms of tenofovir nephrotoxicity are known. Atazanavir can, equally to indinavir, cause urolithiasis, but both drugs have also been associated with chronic kidney disease (CKD) and fast declining eGFR in persons without clinical symptoms of urolithiasis, especially when the plasma drug concentration is boosted by concomitant ritonavir use. In 2012, only a minority of HIV-positive individuals are affected by drug-induced nephrotoxicity. However, in the future, the clinical impact and hence the requirement for more research in this area will likely increase due to ageing and continued cART exposure of the HIV-positive population.


Revolutionizing Healthcare with ChatGPT: An Early Exploration of an AI Language Model's Impact on Medicine at Large and its Role in Pediatric Surgery.
ChatGPT, a natural language processing model, has shown great promise in revolutionizing the field of medicine. This paper presents a comprehensive evaluation of the transformative potential of OpenAI's ChatGPT on healthcare and scientific research, with an exploration on its prospective capacity to impact the field of pediatric surgery. Through an extensive review of the literature, we illuminate ChatGPT's applications in clinical healthcare and medical research while presenting the ethical considerations surrounding its use. Our review reveals the exciting work done so far evaluating the numerous potential uses of ChatGPT in clinical medicine and medical research, but it also shows that significant research and advancements in natural language processing models are still needed. ChatGPT has immense promise in transforming how we provide healthcare and how we conduct research. Currently, further robust research on the safety, effectiveness, and ethical considerations of ChatGPT is greatly needed. V.


Career advising in family medicine: a theoretical framework for structuring the medical student/faculty advisor interview.
There are unique challenges to recruiting students into the specialty of family medicine within academic medical centers. At Virginia Commonwealth University, we developed an advising framework to help students address institutional and personal obstacles to choosing family medicine as a career. The role of a faculty advisor is not to direct the student to a career choice but rather to foster a mentor relationship and help the student come to his or her own realizations regarding career choice. The faculty advisor/medical student interview is conceptualized as five discussion topics: self-knowledge, perception, organizational voice, cognitive dissonance, and anticipatory counseling. This framework is intended to assist faculty in their efforts to encourage students to consider a career in family medicine.


Effect of inhaled disodium cromoglycate and albuterol on energy cost of running in asthmatic children.
The purpose of this study is to evaluate the effect of disodium cromoglycate and albuterol on energy cost of running, gas exchange, and ventilation during maximal exercise in children with exercise-induced asthma (EIA). Twelve children (7.1-15.5 years old) with a history of mild to moderate asthma and EIA performed three maximal exercise tests on a treadmill: 1) test A, without premedication; 2) test B, after premedication with inhaled disodium cromoglycate (DSCG) (40 mg); 3) test C, after premedication with inhaled albuterol (200 micrograms). The energy cost of running was calculated at each minute of exercise. None of the children were limited by dyspnea during the run. The post-exercise fall in FEVi after test A was greater than 20% for each child, the mean fall being 32.8 +/- 11.6%, in comparison with 12.6 +/- 8.9% after test B (P less than 0.001) and 2.5 +/- 5.3% after test C (P less than 0.001). There was no difference in the baseline oxygen uptake for the three tests. Maximum oxygen uptake (VO2 peak) decreased from 43.9 +/- 7.7 mL/min/kg in test A to 37.7 +/- 6.0 mL/min/kg in test B (P less than 0.01) and 39.1 +/- 7.2 mL/min/kg in test C (P less than 0.05). Ventilatory anaerobic threshold in tests B and C was significantly lower than in test A (P less than 0.01). Ventilation (L/min) and energy cost of running (O2 mL/kg/m) were significantly lower in tests B and C than in test A at comparable times. Running time was longer in B and C (P less than 0.05) with respect to A.(ABSTRACT TRUNCATED AT 250 WORDS)


Directed assembly of cylinder-forming block copolymer films and thermochemically induced cylinder to sphere transition: a hierarchical route to linear arrays of nanodots.
A morphological transition from cylinders to spheres was induced in an asymmetric diblock copolymer, poly(styrene)-block-poly(tert-butyl acrylate) (PS-b-PtBA). The periodic arrays of the poly(tert-butyl acrylate) (PtBA) domains were transformed to the ordered poly(acrylic anhydride) (PAA) spheres via the thermal deprotection of tert-butyl acrylate linkages and the subsequent volume change of the minority block. Coupled with techniques to direct the assembly of cylinder-forming block copolymers, this finding provides new routes to fabricate ordered geometries of nanodot arrays.


Flow cytometric DNA analysis of stage I endometrial carcinoma.
Flow cytometric DNA analysis was performed on 203 paraffin-embedded archival specimens obtained from patients with surgical stage I endometrial carcinoma. Primary therapy for those patients (1979-1983) had been definitive extirpation with adjuvant therapy determined by histologic grade, histologic subtype, myometrial invasion, and peritoneal cytologic findings. Diploid DNA patterns were identified in 171 (84%) specimens and nondiploid characteristics were observed in the remaining 32 (25 DNA aneuploid, 7 DNA tetraploid). Although DNA nondiploid specimens accounted for only 16% of all stage I patients, they accounted for 50% of all relapses. Regardless of treatment or other pathologic features, progression-free 5-year Kaplan-Meier survival estimates were 92 and 63% for patients with DNA diploid and DNA non-diploid patterns, respectively (P less than 0.001). Overall 5-year progression-free survival for patients with grade 1 or 2 lesions was 90%; stratification by DNA diploid and DNA nondiploid patterns revealed progression-free survivals of 94 and 64%, respectively (P less than 0.001). Peritoneal cytologic study was positive in seven patients; none of the five with a DNA diploid pattern had a relapse and both with the DNA nondiploid pattern had relapses. These studies suggest that DNA ploidy status may be an objective prognostic determinant for patients with stage I endometrial carcinoma.


Alterations in photosynthetic pigments and amino acid composition of D1 protein change energy distribution in photosystem II.
The marine cyanobacterium Prochlorococcus marinus accumulates divinyl chlorophylls instead of monovinyl chlorophylls to harvest light energy. As well as this difference in its chromophore composition, some amino acid residues in its photosystem II D1 protein were different from the conserved amino acid residues in other photosynthetic organisms. We examined PSII complexes isolated from mutants of Synechocystis sp. PCC 6803, in which chromophore and D1 protein were altered (Hisashi Ito and Ayumi Tanaka, 2011) to clarify the effects of chromophores/D1 protein composition on the excitation energy distribution. We prepared the mutants accumulating divinyl chlorophyll (DV mutant). The amino acid residues of V205 and G282 in the D1 protein were substituted with M205 and C282 in the DV mutant to mimic Prochlorococcus D1 protein (DV-V205M/G282C mutant). Isolated PSII complexes were analyzed by time-resolved fluorescence spectroscopy. Energy transfer in CP47 was interrupted in PSII containing divinyl chlorophylls. The V205M/G282C mutation did not recover the energy transfer pathway in CP47, instead, the mutation allowed the excitation energy transfer from CP43 to CP47, which neighbors in the PSII dimer. Mutual orientation of the subcomplexes of PSII might be affected by the substitution. The changes of the energy transfer pathways would reduce energy transfer from antennae to the PSII reaction center, and allow Prochlorococcus to acquire light tolerance.


Obligatory free radical intermediate in the oxidative activation of the carcinogen N-hydroxy-2-acetylaminofluorene.
We have demonstrated that the nitroxyl free radical form of the carcinogen N-hydroxy-2-acetylaminofluorene (OH-AAF) is an obligatory intermediate in the cumene hydroperoxide-hematin-induced oxidative activation of this carcinogen into 2-nitrosofluorene and N-acetoxy-2-acetylaminofluorene. Both the rate of N-OH-acetylaminofluorene oxidation and the amount of its nitroxyl free radical were experimentally observed as a function of reaction time. Rate equations were derived for a model in which the nitroxyl free radical form of OH-AAF was an obligatory intermediate in the reaction. Using this theory it was possible to compute one experimental variable, the rate of OH-AAF oxidation, utilizing the other experimental variable, the amount of nitroxyl free radical present at any time during the reaction. The theory also predicts a linear relationship between the rate of OH-AAF oxidation and the square of the free radical content; and this was found to be true experimentally. The dismutation rate constant of the nitroxyl free radical of OH-AAF was found to be 2.7-10(5) M-1-s-1.


The Impact of COVID-19 Surge on Clinical Palliative Care: A Descriptive Study From a New York Hospital System.
In spring 2020, New York experienced a surge of patients hospitalized with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or COVID-19) disease, as part of a global pandemic. There are limited data on populations of COVID-19-infected patients seen by palliative care services. To describe a palliative care population at one New York hospital system during the initial pandemic surge. This repeated cross-sectional, observational study collected data on palliative care patients in a large health system seen during the COVID-19 outbreak and compared it with pre-COVID data. Palliative service volume surged from 678 (4% of total admissions) before COVID-19 to 1071 (10% of total admissions) during the COVID-19 outbreak. During the outbreak, 695 (64.9%) of the total palliative patients tested positive for the virus. Compared with a preoutbreak group, this COVID-19-positive group had higher rates of male (60.7% vs. 48.6%, P < 0.01) and Latino (21.3% vs. 13.3%; P < 0.01) patients and less white patients (21.3% vs. 13.3%; P < 0.01). Our patients with COVID-19 also had greater prevalence of obesity and diabetes and lower rates of end-stage organ disease and cancers. The COVID-19-positive group had a higher rate of intensive care unit admissions (58.9% vs. 33.9%; P < 0.01) and in-hospital mortality rate (57.4% vs. 13.1%; P < 0.01) than the preoutbreak group. There was increased odds of mortality in palliative care patients who were COVID-19 positive (odds ratio = 3.21; 95% confidence interval = 2.43-4.24) and those admitted to the intensive care unit (odds ratio = 1.45; 95% confidence interval = 1.11-1.9). During the initial surge of the COVID-19 pandemic in New York, palliative care services experienced a large surge of patients who tended to be healthier at baseline and more acutely ill at the time of admission than pre-COVID-19 palliative patients.


Enhanced Dissolution of a Porous Carrier-Containing Ternary Amorphous Solid Dispersion System Prepared by a Hot Melt Method.
The focus of our study was to employ a solvent-free, thermal process to evaluate the use of a porous carrier in a drug-polymer-porous carrier ternary formulation containing a high drug load (e.g., ≥50% w/w). The purpose of the study was to improve the dissolution properties of the biopharmaceutical classification system class II drug, indomethacin, in the ternary formulation. The effect that the selected polymer has on properties of the formulation was studied, and the formulation characteristics of hypromellose (AF15), copovidone (VA64), and polyvinyl alcohol-polyethylene glycol graft copolymer was evaluated to understand differences in dissolution rates and drug adsorption onto the porous carrier. The ternary formulations were manufactured using a thermal technique that relied on heating and mixing, without the necessity of mechanical shear. All thermally processed granules that employed the porous carrier exhibited immediate release compared with crystalline indomethacin and physical mixtures. In addition, the ternary formulations maintained supersaturation compared with the binary formulations without polymer. The results of this study indicated that the thermally processed ternary formulations containing a porous carrier demonstrated a much improved dissolution profile in nonsink conditions.


Beta-human chorionic gonadotrophin levels before and after the development of pre-eclampsia.
Maternal serum levels of beta-human chorionic gonadotrophin (beta-hCG) concentrations were significantly higher in 41 primigravid patients with proteinuric pre-eclampsia compared with those in 41 primigravid normotensive women matched for age and gestation. beta-hCG was also measured in serial serum samples obtained from 13 women who had a high weight-gain between 20 and 30 weeks of pregnancy. Seven of these pregnancies were subsequently complicated with pre-eclampsia while the remaining six continued to be normotensive; beta-hCG levels were found to rise before the clinical signs of pre-eclampsia appeared.


CYP2C19 loss-of-function alleles are not associated with higher prevalence of gastrointestinal bleeds in those who have been prescribed antidepressants: Analysis in a British-South Asian cohort.
CYP2C19 is a hepatic enzyme involved in the metabolism of antidepressants associated with increased gastrointestinal bleed (GIB) risk. The aim of our study was to explore a possible association between loss-of-function CYP2C19 genotypes and GIB in South Asian ancestry participants prescribed antidepressants. Genes & Health participants with a record in Barts Health NHS Trust (N 22 753) were studied using a cross-sectional approach. CYP2C19 diplotypes were assessed and metabolizer type inferred from consortia guidance. Fisher's exact test was used to compare the prevalence of GIB in different metabolizer categories. Multivariable regression was used to test for association between antidepressant prescriptions and GIB, and between CYP2C19 metabolizer state and GIB in the subcohort prescribed antidepressants. Antidepressants were frequently prescribed (47%, N = 10 612). A total of 864 participants (4%) had a GIB; 534 (62%) had been prescribed a CYP2C19 metabolized antidepressant. There was an independent association between antidepressant prescriptions and GIB events (odds ratio 1.8, confidence interval 1.5-2.0, P < 0.0001). There was no relationship between CYP2C19 inferred poor (P 0.56) or intermediate (P 0.53) metabolizer status and GIB in those prescribed an antidepressant in unadjusted analysis. A multivariable logistic regression model did not show an independent association between poor (P 0.54) or intermediate (P 0.62) CYP2C19 metabolizers and GIB in the subcohort prescribed antidepressants. CYP2C19 dependent antidepressants are associated with increased GIB prevalence. GIB appeared independent from CYP2C19 metabolizer genotype in individuals who had been prescribed antidepressants. Precision dosing based on CYP2C19 genetic information alone is unlikely to reduce GIB prevalence.


Establishment and characterization of a human T-lymphocyte cell line immortalized by SV40 and with abnormal expression of TCR/CD3.
Human lymphocytes derived from the peripheral blood of a healthy woman were transfected with a plasmid carrying the simian virus 40 (SV40) large T antigen. The successfully transformed cells contained SV40 large T DNA and were negative for Epstein-Barr virus (EBV) and human T-cell leukaemia virus (HTLV)-1 genomes. The immortalized cell line was assigned to the T-lymphocyte lineage on the basis of morphological, immunological and cytochemical criteria. While the cells expressed CD1a and CD4 at the cell surface, the CD3 complex was solely intracytoplasmic. Immunoprecipitation studies indicated that these cells lacked T-cell receptor (TCR) alpha-chains but not beta-chains. They were negative for activation markers such as CD25, CD69 and major histocompatibility (MHC) class II molecules. In addition, the transformed cells exhibited a complete growth independency towards interleukin-2 (IL-2). However, after phorbol ester stimulation, CD25 and CD69 markers were expressed and IL-2 was secreted. This new human immortalized T-lymphocytic cell line, which is cell-surface TCR/CD3-negative, may be useful as an in vitro model for studying TCR/CD3 assembly, expression and signal transduction.


Interventions to support family caregivers of people with advanced dementia at the end of life in nursing homes: A mixed-methods systematic review.
Most people with dementia transition into nursing homes as their disease progresses. Their family caregivers often continue to be involved in their relative's care and experience high level of strain at the end of life. To gather and synthesize information on interventions to support family caregivers of people with advanced dementia at the end of life in nursing homes and provide a set of recommendations for practice. Mixed-Methods Systematic Review (PROSPERO no. CRD42020217854) with convergent integrated approach. Five electronic databases were searched from inception in November 2020. Published qualitative, quantitative, and mixed-method studies of interventions to support family caregivers of people with advanced dementia at the end of life in nursing home were included. No language or temporal limits were applied. In all, 11 studies met the inclusion criteria. Data synthesis resulted in three integrated findings: (i) healthcare professionals should engage family caregivers in ongoing dialog and provide adequate time and space for sensitive discussions; (ii) end-of-life discussions should be face-to-face and supported by written information whose timing of supply may vary according to family caregivers' preferences and the organizational policies and cultural context; and (iii) family caregivers should be provided structured psychoeducational programs tailored to their specific needs and/or regular family meetings about dementia care at the end of life. The findings provide useful information on which interventions may benefit family caregivers of people with advanced dementia at the end of life and where, when, and how they should be provided.


Acute effects of short-term subcutaneous terbutaline on theophylline disposition.
Theophylline and subcutaneous terbutaline are frequently used concurrently in the management of acute asthma. Recent evidence demonstrating a reduction in theophylline serum concentrations during concomitant oral terbutaline therapy prompted our evaluation of subcutaneous terbutaline's effect on theophylline pharmacokinetics. Using a randomized, placebo controlled, crossover design, the disposition of a single oral theophylline dose (7 mg/kg) was studied in eight healthy, adult males before and after repeated subcutaneous administration of terbutaline (0.25 mg). Two-way analysis of variance revealed no significant difference in elimination rate constant (ke), area under the concentration-time curve (AUC), or apparent oral clearance (CL/F) of theophylline following terbutaline administration. These results indicate that subcutaneous administration of terbutaline does not alter the pharmacokinetics of single, oral doses of theophylline in adults.