ASHP national survey of pharmacy practice in hospital settings: Impact of COVID-19 pandemic on pharmacy operations-2020.
Results of the 2020 ASHP national survey of pharmacy practice in hospital settings pertaining to pharmacy operational changes implemented in response to the coronavirus disease 2019 (COVID-19) pandemic are presented. Pharmacy directors at 1,437 general and children's medical/surgical hospitals in the United States were surveyed using a mixed-mode method of contact by email and mail. Survey completion was online. The response rate was 18.7%. Seventy-three percent of hospitals implemented changes to hospital units, including 46% that increased intensive care unit bed capacity; 94% made changes to pharmacy supply chain acquisition, changes to products, and/or increased inventory. Staffing changes were implemented by 69% of hospitals, with the most common being staffing reductions (55%) and salary reductions (16%). Medication-use changes were implemented by 86% of hospitals, with treatment guidelines for COVID-19 treatment (79%) and opening compassionate use or investigational drug studies (55%) being the most common. Changes in sterile compounding processes were implemented by 84% of hospitals. Personal protective equipment (PPE) shortages led to 71% of hospitals modifying PPE use standards in sterile compounding. Eighty-seven percent of hospitals changed operational activities, such as changing medication return practices (56%), medication reconciliation processes (46%), intravenous medication recycling (38%), and discharge counseling (37%). Hospitals experienced shortages of many medications, including albuterol inhalers (60%), sedatives and anesthetic agents (58%), neuromuscular blockers (43%), corticosteroids (34%), cardiovascular agents (24%), investigational agents (24%), and dialysis solutions (6%). The pharmacy profession responded to myriad threats to operations and patient care during the COVID-19 pandemic in 2020.


Child disability in a changing world: a human rights approach
The COVID-19 pandemic is pronounced as a turning point in our professional health practice, from which new opportunities will arise with which we must promote the rights of children and people with disabilities.


Retinal vein thrombosis in a patient with pernicious anemia and anticardiolipin antibodies.
A 45-year-old man with pernicious anemia presented with sudden loss of vision due to central retinal vein thrombosis, and was found to have anticardiolipin antibodies and a clotting disorder consistent with the presence of a lupus anticoagulant. He was treated with low dose aspirin and has remained free of recurrent thrombosis over a period of one year. The association between lupus anticoagulant and pernicious anemia is rare, having been reported in only one prior case.


A comparison of pregnancy outcome of emergency modified transvaginal cervicoisthmic cerclage performed in twin and singleton pregnancies.
To compare the pregnancy outcome of emergency cerclage performed in twin and singleton pregnancies. A retrospective trial was carried out in the Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital in Zhejiang, China, from January 2013 to December 2018. A total of 89 women with acute cervical incompetence underwent the emergency cerclage. Main outcome measures were gestational age at delivery and the fetal outcome. There were 31 twin and 58 singleton pregnancies that underwent emergency cerclage. In singleton pregnancies, gestational age at delivery (31.57 ± 6.11 vs 28.81 ± 4.34, p = 0.016), and prolongation of pregnancy (8.69 ± 6.13 vs 5.41 ± 3.72, p < 0.001) were greater, and the preterm delivery rate before 34 weeks was lower (53.4% vs 80.6%, p = 0.011) compared with twin pregnancies. Fetal weight was greater in singletons than in twins (2377.3 ± 862.1 vs 1630.3 ± 557.6, p < 0.001), but the fetal survival rate was similar. The proportion of twin gestations was significantly higher in the group with preterm delivery before 34 weeks (44.6% vs 18.2%, p = 0.011) and the proportion of previous term delivery was higher in the group with delivery at or after 34 weeks (30.3% vs 12.5%, p = 0.039). The group with preterm delivery before 34 weeks had a significantly larger size of visible membranes at cerclage than the group with delivery at or after 34 weeks (2.23 ± 0.86 vs 1.64 ± 0.80, p = 0.001). In binary logistic regression analysis, twin gestations and a larger size of visible membranes were associated with an increased risk of preterm delivery. The pregnancy outcome of emergency cerclage appears to have a better prognosis in singleton pregnancies than in twin pregnancies, and twin pregnancy is a risk factor for preterm delivery.


Effects of Two Commercial Diets on Several Reproductive Parameters in Bitches: Note One-From Estrous Cycle to Parturition.
The close link between nutrition management and reproductive efficiency is well known, but there is very little data available concerning this topic in canine species. The present study aimed to compare the effect of two different diets upon the follicular period and gestation in bitches. Eighteen pluriparus medium and large size bitches were recruited and divided into control (CTR) and experimental (EX) groups and fed, respectively, with a commercial kibble diet and a specially formulated diet from two months before the expected onset of proestrus up to the end of the trial. It was possible to observe how the EX group had a better clinical presentation of the estrous phase, a higher number of ovarian follicles (p < 0.05), a lower percentage of fetal resorption (p < 0.05), and lower oxidative status, expressed by d-ROMs (p < 0.01), at the moment of pregnancy diagnosis compared to CTR group. Moreover, the EX group showed a lower fetal resorption rate and higher litter size (p < 0.05). These results highlight how a diet characterized by high protein and fat content and richer in essential fatty acids can improve reproductive performance in dogs.


Quantitative evaluation of bone-resorptive lesion volume in osteonecrosis of the femoral head using micro-computed tomography.
Objectives To quantify the volume of bone-resorptive lesions in post-collapse osteonecrosis of the femoral head (ONFH) using micro-computed tomography (micro-CT) and assess their characteristics in post-collapse ONFH. Methods We investigated 35 femoral heads resected from 35 patients with ONFH (20 men and 15 women; mean age, 47.2 years). On each of seven coronal high-resolution micro-CT slices of the femoral head, the bone-resorptive areas were extracted using bone microstructure measurement software. Next, the total bone-resorptive volume ratio, defined as the ratio of all bone-resorptive cross-sectional areas to all femoral head cross-sectional areas in all seven slices, was calculated. Associations between total bone-resorptive volume ratio and sex, age, ONFH-associated factors, patient workload levels, ONFH stage, ONFH type, necrotic volume on magnetic resonance imaging, and duration from the onset of pain to surgery were analyzed. Lesion location and the association between bone-resorptive lesion and collapse were also evaluated. Results The mean total bone-resorptive volume ratio was 7.0±6.0%, which varied significantly by ONFH stage (ARCO collapse quantitation 3A, 3.5±2.1%; 3B, 6.8±3.0%; and 3C, 13.6±8.8%). ONFH stage was independently associated with total bone-resorptive volume ratio (P<0.05). Furthermore, high bone-resorptive volume ratios were found in the anterior femoral head and were associated with collapse. Conclusions This study demonstrated that bone-resorptive volume in post-collapse ONFH was significantly associated with the disease stage, which was more widespread in the anterior portion of the femoral head than in the posterior portion.


Contribution to the study of live streptomycin-dependent Salmonella vaccines: the problem of reversion to a virulent form.
The recovery of virulence by means of reversion of a live streptomycin-dependent (Sm D) Salmonella typhimurium vaccine was studied in CD-1 Swiss mice. Initially, a one-step Sm D mutant was obtained from a virulent streptomycin-sensitive (Sm S) S. typhimurium strain. Afterwards, two pools of streptomycin-independent (Sm I) revertants were prepared from the Sm D strain. The virulence of the Sm D strain and of the Sm I revertants was tested intraperitoneally. In the virulence testing the original suspension of the Sm I revertants, as well as their 1st and 10th passages on plain medium, medium+50 mug. streptomycin/ml. and medium+1000 mug. streptomycin/ml. were used. The results show that the Sm D mutant was avirulent, its avirulence being due to an intrinsic, genetic quality. The Sm I revertants, compared to the original Sm S strain, also displayed a lack of virulence. However, afterwards, the Sm I revertants behaved quite differently, according to their subsequent passages. Indeed, there was an increase in virulence after passages on plain medium, whereas similar passages on medium containing the drug, the virulence not only failed to increase, but disappeared almost completely. Moreover, the passages on medium containing 1000 mug. streptomycin/ml. induced a return to the status of drug-dependence. The danger of recovery of virulence by means of revertants is evaluated.


A technical note on the assessment of human sperm vitality using eosin-nigrosin staining.
How many spermatozoa and slides need to be counted to make a reliable assessment of sperm vitality? Currently, various authorities recommend assessing human sperm vitality on counts of at least 200 cells, but on one or two slides. This was an observational study on duplicate eosin-nigrosin stained sperm vitality slides made from 58 ejaculates. Assessments were made using counts of 2 × 100 and 1 × 200 cells per slide, all performed by the same trained expert observer. Although assessments tend to show fewer and smaller outlier values when based on counts of 200 spermatozoa than 100, and on 2 × 200 than 1 × 200, counting 200 spermatozoa from one slide provides a result with sufficient accuracy for the clinical purpose of ascertaining whether the immotile spermatozoa in an ejaculate showing low sperm motility are alive or dead. While the increased accuracy of results derived from counts of 2 × 200 cells might be important in research studies where sperm vitality is the specific end-point of interest, counting at least 200 spermatozoa from one smear is sufficiently accurate for the clinical purpose of establishing whether the immotile spermatozoa seen in ejaculates with low sperm motility are alive or not. Consequently, the extra workload of performing replicate counts and the associated calculations does not increase the clinical value of the result, and hence is unnecessary in routine semen analysis. Careful laboratory technique as well as proper staff training and competence are essential. The conclusions might not be applicable to staining methods other than the recommended one-step eosin-nigrosin technique.


The ovaries of transgender men indicate effects of high dose testosterone on the primordial and early growing follicle pool.
Androgens are essential in normal ovarian function and follicle health but hyperandrogenism, as seen in polycystic ovary syndrome, is associated with disordered follicle development. There are few data on the effect of long-term exposure to high levels of testosterone as found in transgender men receiving gender-affirming endocrine therapy. In this study, we investigate the effect of testosterone on the development, morphological health and DNA damage and repair capacity of human ovarian follicles in vivo and their survival in vitro. Whole ovaries were obtained from transgender men (mean age: 27.6 ± 1.7 years; range 20-34 years, n = 8) at oophorectomy taking pre-operative testosterone therapy. This was compared to cortical biopsies from age-matched healthy women obtained at caesarean section (mean age: 31.8±1.5 years; range= 25-35 years, n=8). Cortical tissues were dissected into fragments and either immediately fixed for histological analysis or cultured for 6 days and subsequently fixed. Follicle classification and morphological health were evaluated from histological sections stained with H&E and expression of γH2AX as a marker of DNA damage by IHC. In uncultured tissue, testosterone exposure was associated with reduced follicle growth activation, poor follicle health and increased DNA damage. After 6 days of culture, there was enhanced follicle activation compared to control with further deterioration in morphological health and increased DNA damage. These data indicate that high circulating concentrations of testosterone have effects on the primordial and small-growing follicles of the ovary. These results may have implications for transgender men receiving gender-affirming therapy prior to considering pregnancy or fertility preservation measures.


Insoluble matrix proteins from shell waste for synthesis of visible-light response photocatalyst to mineralize indoor gaseous formaldehyde.
HCHO is the most concerned indoor air pollutant that photocatalytic degradation is a feasible approach. To achieve efficient and complete degradation of HCHO under visible light irradiation, heteroatoms are usually doped in TiO2. But using natural materials as a dopant instead of expensive and toxic chemicals to fertilize TiO2 remains challenging. This paper proposes a sustainable and green approach to synthesize an efficient N, Ca co-doped TiO2 photocatalyst (TIMP) by using the insoluble matrix proteins (IMPs) extracted from abalone shell. TIMP-0.8 achieves near completely degradation HCHO within 45 min under visible light at ambient temperature and exhibits superior stability after 7 cycles. TIMP-0.8 has monodispersity with smaller diameter, high porosity, abundant defects and high adsorption affinity for surface hydroxyls compared with pure TiO2. With the assistance of IMPs, the rate-determining step of HCHO degradation changes from -COOH oxidation to spontaneous decomposition of HCO3-, significantly facilitating the elimination and mineralization of HCHO. Overall, IMPs from abalone shell are natural surfactant, bio-templet, and dopant for TiO2 modification, contributing to desirable visible-light photocatalytic performance for HCHO degradation. This paper provides new insight for high-value utilization of waste shell and photocatalytic indoor purification.


The choleretic role of tauroursodeoxycholic acid exacerbates alpha-naphthylisothiocyanate induced cholestatic liver injury through the FXR/BSEP pathway.
The aim of this study was to determine the effect of tauroursodeoxycholic acid (TUDCA) on the alpha-naphthylisothiocyanate (ANIT)-induced model of cholestasis in mice. Wild-type and farnesoid X receptor (FXR)-deficient (Fxr-/- ) mice were used to generate cholestasis models by gavage with ANIT. Obeticholic acid (OCA) was used as a positive control. In wild-type mice, treatment with TUDCA for 7 days resulted in a dramatic increase in serum levels of alanine aminotransferase (ALT), with aggravation of bile infarcts and hepatocyte necrosis with ANIT-induction. TUDCA activated FXR to upregulate the expression of bile salt export pump (BSEP), increasing bile acids (BAs)-dependent bile flow, but aggravating cholestatic liver injury when bile ducts were obstructed resulting from ANIT. In contrast, TUDCA improved the liver pathology and decreased serum ALT and alkaline phosphatase (ALP) levels in ANIT-induced Fxr-/- mice. Furthermore, TUDCA inhibited the expression of cleaved caspase-3 and reduced the area of terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining in the model mice. TUDCA also upregulated anion exchanger 2 (AE2) protein expression, protecting cholangiocytes against excessive toxic BAs. Our results showed that TUDCA aggravated cholestatic liver injury via the FXR/BSEP pathway when bile ducts were obstructed, although TUDCA inhibited apoptotic activity and protected cholangiocytes against excessive toxic BAs.


New approach to studies on ECG dynamics: extraction and analyses of QRS complex irregularity time series.
How to extract information intensively from ECGs for the diagnosis of cardiovascular diseases and assessment of heart function is a topical subject. Using a method based on the wavelet transform to calculate the irregularity of the QRS complex, which may relate to inotropy, the QRS complex irregularity time series is successfully extracted from original ECG signals. This provides a new approach to studies of ECG dynamics. With the help of non-linear dynamics theory, the QRS complex irregularity time series of eight subjects from the MIT/BIH arrhythmia database are studied qualitatively and quantitatively, and the characteristics of ECG dynamics are analysed extensively. The power spectrum, phase portrait, correlation dimension, largest Lyapunov exponent, time-dependent divergence exponent and complexity measure all verify the fact that ECG dynamics are dominated by an underlying 5-6-dimensional non-linear chaotic system, whose complexity measure is about 0.7. The QRS complex irregularity time series contains abundant information about all parts of the heart and the regulation of the autonomic nervous system, and so further analyses are of great potential theoretical and clinical significance to patho-physiology studies and ambulatory monitoring.


Association Between Diet Quality and Cardiometabolic Risk Factor Clustering Stratified by Socioeconomic Status Among Chinese Children.
Few studies have evaluated the long-term relationship between diet quality and cardiometabolic risk factor clustering among children. The moderating effect of socio-economic status (SES) is of interest. To investigate the association between diet quality with cardiometabolic risk among Chinese children and to explore the moderating effect of SES. In this cohort study, 5 waves (1997-2009) of the China Health and Nutrition Survey were used. Diet quality was measured by a modified version of the Chinese Children Dietary Index (mCCDI) based on Dietary Guidelines for Chinese. Children between the ages of 7 and 17 (n = 2903) who completed at least 2 surveys were included. Those who missed measures or had hypertension or diabetes at baseline were excluded. The fasting blood samples were collected in 2009. Waist circumference (WC) and blood pressure (BP) were measured in each survey. A continuous cardiometabolic risk score (MetScore) was derived by a confirmatory factor analysis of 5 components: WC, BP, glucose, triglycerides, and high-density lipoprotein cholesterol. Considering the latency period of the effect of behaviors, the mCCDI was lagged by the period between surveys. Linear regression was used to analyze the association of mCCDI with MetScore and its components. Mixed effect linear regression and lagged mCCDI were used for WC and BP models. Higher mCCDI was independently associated with a lower MetScore at follow-up (β: -.11; 95% CI: -.18 to -.04). Higher lagged mCCDI over time was associated with a lower WC z score overall (β: -.05; 95% CI: -.08 to -.01) and among children in the low SES group (β: -.09; 95% CI: -.14 to -.04) but not those in the high SES group. When examining the 15 mCDDI components separately, scores for 5 components: more grains, vegetables, soybeans and its products; less sugar-sweetened beverages; and more diet variety were significantly associated with a lower MetScore. Among Chinese children, higher diet quality measured by mCCDI was independently associated with a lower MetScore at follow-up.


MR Measures of Small Bowel Wall T2 Are Associated With Increased Permeability.
Increased small bowel permeability leads to bacterial translocation, associated with significant morbidity and mortality. Biomarkers are needed to evaluate these changes in vivo, stratify an individual's risk, and evaluate the efficacy of interventions. MRI is an established biomarker of small bowel inflammation. To characterize changes in the small bowel with quantitative MRI measures associated with increased permeability induced by indomethacin. Prospective single-center, double-blind, two-way crossover provocation study. A provocation cohort (22 healthy volunteers) and intrasubject reproducibility cohort (8 healthy volunteers). 2D balanced turbo field echo sequences to measure small bowel wall thickness, T2 , and motility acquired at 3T. Participants were randomized to receive indomethacin or placebo prior to assessment. After a minimum 2-week washout, measures were repeated with the alternative allocation. MR measures (wall thickness, T2 , motility) at each study visit were compared to the reference standard 2-hour lactulose/mannitol urinary excretion ratio (LMR) test performed by a lab technician. All analyses were performed blind. Normality was tested (Shapiro-Wilk's test). Paired testing (Student's t-test or Wilcoxon) determined the significance of paired differences with indomethacin provocation. Pearson's correlation coefficient compared significant measures with indomethacin provocation to LMR. Intrasubject (intraclass correlation) and interrater variability (Bland-Altman) were assessed. Indomethacin provocation induced a significant increase in LMR compared to placebo (P < 0.05) and a significant increase in small bowel T2 (0.12 seconds compared to placebo 0.07 seconds, P < 0.05). Small bowel wall thickness (P = 0.17) and motility (P = 0.149) showed no significant change. T2 and LMR were positively correlated (r = 0.68, P < 0.05). T2 measurements were robust to interobserver (intraclass correlation 0.89) and intrasubject variability (Bland-Altman bias of 0.005 seconds, 95% confidence interval [CI] -0.04 to +0.05 seconds, and 0.0006 seconds, 95% CI -0.05 to +0.06 seconds). MR measures of small bowel wall T2 were significantly increased following indomethacin provocation and correlated with 2-hour LMR test results. 1 TECHNICAL EFFICACY STAGE: 2.


The role of endoscopic retrograde cholangiopancreatography in management of patients recovering from acute biliary pancreatitis in the laparoscopic era.
Traditionally an episode of acute biliary pancreatitis (ABP) is an indication for direct imaging of the biliary tree. The optimal approach may vary according to local expertise, and endoscopic retrograde cholangiopancreatography (ERCP) is the most common. The fact that the incidence of choledocholithiasis in patients recovering from ABP varies between 3 and 33% raises a question about the necessity of visualizing the biliary tree in all patients recovering from ABP. In order to evaluate this policy, we reviewed 48 ERCPs performed on patients recovering from ABP who were scheduled for laparoscopic cholecystectomy (LC). We checked the correlations between ERCP findings and the severity of pancreatitis, biochemistry values (which were sampled during the acute phase), and ultrasonographic (US) findings. The ERCP demonstrated common bile duct (CBD) stones in 11 (22.9%) patients. US finding of a dilated CBD and maximal aspartate transaminase (AST) values higher than 90 units/l were significantly correlated with CBD stones (a relative risk [RR] of 2.95 with a 95% confidence interval [CI] for a dilated CBD and RR of 3.89 with a 95% CI of 1.18-12.80 for an AST value higher than 90 units/l). No other parameters were significantly correlated with CBD stones. We, therefore, recommend performing a preoperative ERCP only on patients who present with an ultrasonographic finding of CBD dilatation. The correlation to high AST is still to be proven.


Morbidity of tumour enucleation for renal cell carcinoma (RCC): results of a single-centre prospective study.
To evaluate the incidence and risk factors of adverse events (AEs) after tumour enucleation (TE). Between 2006 and 2009, clinical, surgical and functional data were prospectively gathered from 200 consecutive patients who had open TE. TE was done by blunt dissection using the natural cleavage plane between the tumour capsule and normal parenchyma. All the AEs were stratified for severity according to the National Cancer Institute Common Toxicity Criteria version 2.0 grading system. Risk factors for AEs were determined by univariate analysis. The mean (range) tumour size was 3.2 (0.8-10.0) cm, and mean (range) warm ischaemia time (WIT) was 16.5 (5-31) min. Overall, 32 AEs (six grade I, 20 grade II, and six grade III) occurred after TE in 30 patients and of those 27 were surgical (13.5%) and five were medical AEs (2.5%). On univariate analysis, imperative/relative surgical indication was the only predictive factor for the development of medical AEs. Statistically significant predictors of overall surgical AEs and urinary fistula were clinical tumour dimension, completely endorenal tumour growth and a positive tumour relationship with the urinary collecting system (UCS). A completely endorenal tumour growth and a positive tumour relationship with the UCS also correlated with bleeding requiring transfusions. Completely endorenal tumour growth was significantly associated with major (grade III) AEs. The TE technique was associated with a 16% AE rate and of those only 3% required re-intervention (grade III). Urinary fistula occurred in six patients (3%) and ureteric stenting was required in 0.5% of cases.


An optimized technology platform for the rapid multiplex molecular analysis of genetic alterations associated with leukemia.
Molecular methods play a critical role in the accurate diagnosis of leukemia by complementing morphologic, cytochemical, immunophenotypic, and cytogenetic analyses. We developed a multiplex reverse transcription-polymerase chain reaction (RT-PCR) method combined with liquid bead array cytometry for the rapid detection of genetic alterations associated with leukemia. Fusion transcripts corresponding to the most common recurrent chromosomal translocations were reproducibly detected in as low as 0.1-10 ng of total RNA with an analytical sensitivity of 0.01-1%. Multiday, multilot, multioperator, and multi-instrument precision studies, for a total of 678 independent measures in 46 runs, showed a very high reproducibility with 100% agreement among replicates. Using multiplex panels for four to 20 independent targets, we demonstrate the flexibility of the method to codetect rare splicing isoforms, discriminate among multiple variants generated by unique cytogenetic abnormalities, identify distinct chromosomal partners involved with 11q23 or 17q21 rearrangements, and assess cryptic abnormalities not detectable by standard cytogenetics such as the t(12;21), del(1p32), or NPM1 mutations. Overall, three different internal control transcripts and 34 variants resulting from 18 abnormal chromosomal sites were evaluated. These results underscore the value of the multiplex assay system as a sensitive and reliable technology platform for the characterization of relevant genetic alterations in leukemia.


Effects of D1 and D2 dopamine receptor antagonists and catecholamine depleting agents on the locomotor stimulation induced by dizocilpine in mice.
Low doses of the uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (MK-801) induce locomotor stimulation in mice, whereas higher doses are associated with ataxia, stereotyped behaviors and catalepsy. We investigated the role of dopamine receptors and presynaptic dopamine neurons in the locomotor effects of dizocilpine. For comparison, we studied several other drugs that induce locomotor stimulation in mice. Pretreatment of male mice with haloperidol (0.1 mg/kg, i.p.) completely prevented the stimulation of normally coordinated locomotion induced by a non-intoxicating dose of dizocilpine (0.1 mg/kg, i.p.); haloperidol also attenuated the locomotor stimulation produced by phencyclidine (PCP, 1 and 2 mg/kg, i.p.), d-amphetamine (2 and 5 mg/kg, i.p.) and diazepam (0.5 mg/kg, i.p.). Haloperidol (doses up to 2.5 mg/kg) did not attenuate the ataxia and decreased locomotion induced by higher doses of dizocilpine (1 and 2 mg/kg). The active cis isomer of flupenthixol (0.5 mg/kg, i.p.), an antagonist of both D1 and D2 dopamine receptors, also diminished the stimulant actions of all of the test drugs, whereas its inactive trans form did not. The selective D1 antagonist R(+/-)-SCH 23390 (0.1 mg/kg) and the selective D2 antagonist raclopride (1 mg/kg) had little effect on the stimulatory effect of dizocilpine, although they did reduce the stimulation produced by PCP, d-amphetamine and diazepam. However, pretreatment with a combination of R(+/-)SCH 23390 and raclopride completely prevented dizocilpine-induced locomotor stimulation. Pretreatment with alpha-methyl-p-tyrosine (AMPT, 50 and 250 mg/kg), an inhibitor of tyrosine hydroxylase, or with 6-hydroxydopamine (6-OH-DA, 50 micrograms, i.c.v.), a neurotoxin that destroys brain dopaminergic and noradrenergic neurons, did not attenuate the locomotor stimulation induced by dizocilpine, although these treatments did reduce the stimulant effects of d-amphetamine. In AMPT or 6-OH-DA pretreated mice, haloperidol (0.125 mg/kg) prevented the stimulatory effect of dizocilpine. These results support a role for dopamine receptors in the stimulation of normally coordinated locomotion by dizocilpine. However, the locomotor stimulant effect of dizocilpine, unlike that of d-amphetamine, can be expressed in the presence of D1 or D2 dopamine receptor blockade and does not appear to be dependent on intact presynaptic mechanisms.


[Testing of industrial repellents against gnats and horseflies].
According to the Austrian Cosmetic Recommendation a minimum period of protection of 2 hours is to be provided. This applies to the preparation on the market with the substance "Merck 3535" in aqueous concentration in the tested area. However, this protection period was not achieved for horseflies. In test trials the characteristic of the individual nuisance should be taken into account.


Interactive association of maternal education and peer relationship with oppositional defiant disorder: an observational study.
The objectives of this research were to gain insights on the interactive effects, by measuring familial and peer-related risk factors in youths with oppositional defiant disorder (ODD). Participants were college students recruited nationwide, with age between 18 and 25. Through the consensus of expert meetings, a set of questionnaires were used to evaluate the familial status, participant's peer group conditions, high-risk environment of illicit substance use, and oppositional symptoms. The logistic regression was performed to see the independent and interactive risk factors for ODD. A total of 981 subjects were enrolled. Six variables significantly associated with ODD at the multivariate logistic regression, including male, night division, poor academic performance, high risk environment, peer with illicit substance use and high maternal education level. High maternal education exerted independent protective effect on the development of ODD (adjusted odds ratio, aOR = 0.65, 95% CI = 0.44-0.99). Peer with illicit substance use was more likely to associate with ODD in the low maternal education group. The 2-way interactive effect of maternal education and peer with substance use on the development of ODD was OR = 4.96 (2.96, 8.31). The present study highlights the influence of maternal education level to ODD and its interaction with peer of illicit substance use. Our findings imply that the familial attachment and peer interaction are essential stages for the development of human behavior. The research protocol was reviewed and approved by the ethical review committee of National Taiwan University Hospital (number 201505057RINC ) and registered at clinical trial systems at National Taiwan University. In addition, subjects' information was anonymous and de-identified prior to any analysis.


[Detection, on the Escherichia coli chromosome, of the locus necessary to maintaining an autonomous state of sex factor F'lac].
The ability of some Escherichia coli strains to serve as effective recipients in crosses with donor strains carrying various plasmids (F'lac, F'lac gal Mu, RP4) has been studied. The experiments have shown that there is a locus maintaining the autonomous state of the F-factors. The function of this locus named Fpm ("F plasmids maintenance") is dispensable for both a bacterial cell itself and RP4 plasmid existence in the cell.


Open reduction and interfragment wire fixation of comminuted nasal fractures.
Manual reduction is the first choice for all types of acute nasal fractures; open reduction is the choice for fractures that do not respond to manual reduction or complicated fractures at the base of the nasal pyramid. From 1968 to 1986, 564 cases of nasal fractures were treated at the Department of Plastic and Reconstructive Surgery, Jikei University School of Medicine, Tokyo. Of these, 241 were acute nasal fractures, 52 of which were associated with facial bone fractures. Deviated types of fractures that cannot be correctly repositioned by manual reduction and some depressed fractures often require a surgical approach for reduction and fixation. In our series of 241 patients with acute nasal fractures, 43 (18%) were treated by open reduction and fixed with soft stainless steel wires.


The functional scope of plant microRNA-mediated silencing.
Deep sequencing has identified a complex set of plant miRNAs that potentially regulates many target genes of high complementarity. Furthermore, the discovery that many plant miRNAs work through a translational repression mechanism, along with the identification of noncanonical targets, has encouraged bioinformatic searches with less stringent parameters, identifying an even wider range of potential targets. Together, these findings suggest that any given plant miRNA family may regulate a highly diverse set of mRNAs. Here we present evolutionary, genetic, and mechanistic evidence that opposes this idea but instead suggests that families of sequence-related miRNAs regulate very few functionally related targets. We propose that complexities beyond complementarity impact plant miRNA target recognition, possibly explaining the current disparity between bioinformatic prediction and functional evidence.


Chondrocyte viability in fresh and frozen large human osteochondral allografts: effect of cryoprotective agents.
Chondrocyte survival is a major goal for the effective storage and clinical performance of human osteochondral allografts. The majority of animal and human cryopreservation studies conducted so far have been performed in small osteochondral cylinders. Using human tibial plateaus as a model for large osteochondral pieces, this work sought to evaluate the cryoprotective efficiency of glycerol and dimethylsulfoxide (DMSO), and to identify cryopreservation conditions suitable for use in tissue banks. Human tibial plateaus harvested from 7 cadaveric tissue donors were incubated in the presence or absence of cryoprotective agents (CPA): 10% or 15% glycerol and 10% DMSO in a Ham F-12 nutrient mixture. Chondrocyte viability was assessed immediately after thawing, using the MTT reduction assay and a fluorescence microscopic method. The tibial plateaus frozen in the absence of CPA showed a significant decrease in chondrocyte viability. The use of CPA significantly increased chondrocyte viability compared with cartilage frozen without CPA (nearly 50% versus 80% living chondrocytes with 10% glycerol versus 10% DMSO, respectively) relative to that in fresh cartilage. In this regard, 10% DMSO was slightly more effective than either 10% or 15% glycerol, eliciting the recovery of approximately 15% relative to the living chondrocyte content in fresh cartilage. In all conditions, fluorescence microscopic studies showed that surviving chondrocytes were restricted to the superficial cartilage layer. Human tibial plateaus seemed to be a good experimental model to establish cryopreservation methods applicable to large human osteochondral pieces in tissue banks.


Axisymmetric bare freestanding films of highly viscous liquids: Preparation and real-time investigation of capillary leveling.
Thin liquid films are important in many scientific fields. In particular, films with both the surface layers exposed to a different fluid phase, known as freestanding films, are relevant in the ambit of foams and emulsions. Hence, there is a great interest in developing novel techniques allowing to form large and stable freestanding liquid films and to follow their dynamics. We develop a novel opto-mechanical tool allowing to perform and study the preparation and the capillary leveling flow of axisymmetric bare freestanding liquid films. The tool is composed by a customized motorized iris diaphragm and by an innovative joint imaging setup combining digital holography and white light color interferometry that enables real-time measurement of film thickness over a large field of view. The dynamics of films made of a model Newtonian fluid, i.e., high-viscosity silicone oil, is studied. Direct numerical simulations and a hydrodynamic model based on the lubrication theory are used to support the experimental results. Iris opening induces the formation of large circular freestanding films with a stepped profile. Once iris opening is stopped, the films undergo a capillary leveling flow tending to flatten their profile. The leveling flow follows the theoretical scaling given by Ilton et al. [1]. We prove through numerical simulations that an equi-biaxial extensional flow occurs at the film center. Furthermore, we observe the formation and dynamics of dimples in bare freestanding films for the first time.


The immunoelectronmicroscopic localization of human platelet factor 4 in tissue mast cells.
Initial immunohistochemical localization of human platelet factor 4 (PF4) in tissue mast cells suggested that the protein was present in the mast cell granule. It was proposed that this could reflect binding of PF4 to heparin or heparan sulphate, known granule constituents. We report here the confirmation of granule localization by an immunoelectron microscopical method. The possible role of such binding is unknown, but the potential for cationic proteins of platelet origin interacting with vessel wall constituents is discussed.


Time-Resolved Proteomics Extends Ribosome Profiling-Based Measurements of Protein Synthesis Dynamics.
Ribosome profiling is a widespread tool for studying translational dynamics in human cells. Its central assumption is that ribosome footprint density on a transcript quantitatively reflects protein synthesis. Here, we test this assumption using pulsed-SILAC (pSILAC) high-accuracy targeted proteomics. We focus on multiple myeloma cells exposed to bortezomib, a first-line chemotherapy and proteasome inhibitor. In the absence of drug effects, we found that direct measurement of protein synthesis by pSILAC correlated well with indirect measurement of synthesis from ribosome footprint density. This correlation, however, broke down under bortezomib-induced stress. By developing a statistical model integrating longitudinal proteomic and mRNA-sequencing measurements, we found that proteomics could directly detect global alterations in translational rate caused by bortezomib; these changes are not detectable by ribosomal profiling alone. Further, by incorporating pSILAC data into a gene expression model, we predict cell-stress specific proteome remodeling events. These results demonstrate that pSILAC provides an important complement to ribosome profiling in measuring proteome dynamics.


Potentially inappropriate drug use and health-related quality of life in the elderly.
To investigate the relationship between two widely used, generic health-related quality of life (HRQOL) measures, Short Form-12 (SF-12) and EuroQol's EQ-5D, and potentially inappropriate drug use in an elderly cohort. Longitudinal retrospective cohort study. Medical Expenditure Panel Survey, panel 5. Respondents aged 65 years or older. Participants with potentially inappropriate drug use were identified by using National Drug Codes based on the Beers updated criteria. The dependent variable, HRQOL, was measured by using self-reported scores on the SF-12 (physical component summary, mental component summary, and global rating of health) and the EQ-5D (index and visual analog scale). The HRQOL data were regressed onto scores for potentially inappropriate drug, age, sex, number of prescriptions, race, comorbidity, and previous HRQOL. Regression analysis revealed statistically significant models for all five HRQOL equations (adjusted R2=26.50-53.83%, p<0.0001). However, potentially inappropriate drug use was not a significant predictor of HRQOL in any of the models tested. Previous HRQOL, as measured by using the SF-12 (global, physical component summary, and mental component summary) and the EQ-5D (index and visual analog scale), was a significant predictor of HRQOL; number of prescriptions was also a significant predictor of HRQOL, as measured by using the SF-12 (global and physical component summary) and the EQ-5D (index and visual analog scale). The results supported others showing that a significant proportion of the elderly receive care that is potentially harmful and not supported by evidence-based practice. Use of a disease-specific HRQOL scale may be more responsive to measuring the impact of potentially inappropriate drugs on patients' HRQOL.


Patient adherence to physical activity advice (PAPA) in patients with low back pain: Study protocol for a multicentre randomized controlled trial.
It seems that nonspecific low back pain (NSLBP) cannot be successfully treated with a single intervention of any kind. However, a positive effect of an active lifestyle including physical activities in NSLBP is reported on pain and disability. Advising regular physical activity is one of the mainstays in physical therapy treatment, appealing to patients' adherence. Many patients with low back pain do not seem to adhere to their physiotherapist's advice. In this study, the influence of social support on adherence to an activity advice will be investigated. Objective of the study is to investigate if patients with low back pain randomised to receive social support adhere better to an activity advice than patients randomised to the control group. In a randomized controlled clinical trial, in private physiotherapy practices in the Netherlands, patients ≥18 years presenting with at least a second episode of nonspecific low back pain will be randomized over two groups: activity advice and social support as an add-on to usual care versus usual care alone. The primary outcome measure is adherence to an activity advice measured by an activity monitor at 1, 6 and 12 weeks. All other objectives will be measured by questionnaires. This project is the first comprehensive research project to assess whether social support influences adherence to an activity advice outside the clinical setting without supervision of a healthcare provider. Where evidence of influence on social support is lacking, findings may have implications for the management of patients with low back pain. In addition, findings may enable physiotherapists to predict in which patients with low back pain an activity advice with social support makes sense. GOV: NCT02996955.


Characterization of Plasma Immunoglobulin G Responses in Elite Neutralizers of Human Cytomegalovirus.
Human cytomegalovirus (HCMV) is the most common infectious complication of organ transplantation and cause of birth defects worldwide. There are limited therapeutic options and no licensed vaccine to prevent HCMV infection or disease. To inform development of HCMV antibody-based interventions, a previous study identified individuals with potent and broad plasma HCMV-neutralizing activity, termed elite neutralizers (ENs), from a cohort of HCMV-seropositive (SP) blood donors. However, the specificities and functions of plasma antibodies associated with EN status remained undefined. We sought to determine the plasma antibody specificities, breadth, and Fc-mediated antibody effector functions associated with the most potent HCMV-neutralizing responses in plasma from ENs (n = 25) relative to that from SP donors (n = 19). We measured antibody binding against various HCMV strains and glycoprotein targets and evaluated Fc-mediated effector functions, antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP). We demonstrate that ENs have elevated immunoglobulin G binding responses against multiple viral glycoproteins, relative to SP donors. Our study also revealed potent HCMV-specific antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis activity of plasma from ENs. We conclude that antibody responses against multiple glycoprotein specificities may be needed to achieve potent plasma neutralization and that potently HCMV elite-neutralizing plasma antibodies can also mediate polyfunctional responses.


Single-cell analysis highlights a population of Th17-polarized CD4+ naïve T cells showing IL6/JAK3/STAT3 activation in pediatric severe aplastic anemia.
Acquired aplastic anemia (AA) is recognized as an immune-mediated disorder resulting from active destruction of hematopoietic cells in bone marrow (BM) by effector T lymphocytes. Bulk genomic landscape analysis and transcriptomic profiling have contributed to a better understanding of the recurrent cytogenetic abnormalities and immunologic cues associated with the onset of hematopoietic destruction. However, the functional mechanistic determinants underlying the complexity of heterogeneous T lymphocyte populations as well as their correlation with clinical outcomes remain to be elucidated. To uncover dysfunctional mechanisms acting within the heterogeneous marrow-infiltrating immune environment and examine their pathogenic interplay with the hematopoietic stem/progenitor pool, we exploited single-cell mass cytometry for BM mononuclear cells of severe AA (SAA) patients pre- and post-immunosuppressive therapy, in contrast to those of healthy donors. Alignment of BM cellular composition with hematopoietic developmental trajectories revealed potential functional roles for non-canonically activated CD4+ naïve T cells in newly-diagnosed pediatric cases of SAA. Furthermore, single-cell transcriptomic profiling highlighted a population of Th17-polarized CD4+CAMK4+ naïve T cells showing activation of the IL-6/JAK3/STAT3 pathway, while gene signature dissection indicated a predisposition to proinflammatory pathogenesis. Retrospective validation from our SAA cohort of 231 patients revealed high plasma levels of IL-6 as an independent risk factor of delayed hematopoietic response to antithymocyte globulin-based immunosuppressive therapy. Thus, IL-6 warrants further investigation as a putative therapeutic target in SAA.


HLA-C*07:06:01:06-A new HLA-C allele identified in an Ao tribal individual from Nagaland, North-East India.
Two nucleotide changes in the 3'untranslated region of HLA-C*07:06:01:01 results in the novel allele, HLA-C*07:06:01:06.


Association of residential mobility with child health: an analysis of the 2007 National Survey of Children's Health.
To describe the association of residential mobility with child health. We conducted descriptive, bivariate, and multivariable analyses of data from 63,131 children, 6-17 years, from the 2007 National Survey of Children's Health. Logistic regression was used to explore the association of residential mobility with child health and measures of well-being. Analyses were carried out using SAS-callable SUDAAN to appropriately weight estimates and adjust for the complex sampling design. After adjusting for age, race/ethnicity, presence of a special health care need, family structure, parental education, poverty level, and health insurance status, children who moved ≥ 3 times were more likely to have poorer reported overall physical (AOR 1.21 [95 %CI: 1.01-1.46]) and oral health status (AOR 1.31 [95 % CI: 1.15-1.49]), and ≥ 1 moderate/severe chronic conditions (AOR 1.40 [95 % CI: 1.19-1.65]) than children who had no lifetime moves. When compared to children who had never moved, children who moved ≥ 3 times were more likely to be uninsured/have periods of no coverage (AOR 1.35; 95 % CI: 0.98-1.87) and lack a medical home (AOR 1.16, 95 % CI: 1.04-1.31). None of the outcomes were statistically significant for children who moved fewer than 3 times. Clinicians need to be aware that children who move frequently may lack stable medical homes and consistent coverage increasing their risk of poor health outcomes and aggravation of mild or underlying chronic conditions. Public health systems could provide the necessary link between parents and clinicians to ensure that continuous, coordinated care is established for children who move frequently.


Physiologic alteration of fibrinogen levels: influence upon patency of microvenous anastomoses.
High baseline levels of plasma fibrinogen have been correlated with increased risk for cardiovascular disease. Since fibrinogen plays a central role in both coagulation and platelet aggregation and is a primary component of thrombi, this study was designed to assess the relationship of circulating fibrinogen concentration and vein anastomotic patency. Subcutaneously injected turpentine was used to increase, and intravenous ancrod (snake venom) to decrease, plasma fibrinogen levels. Rat femoral vein anastomoses were performed, and patency was assessed after 120 min of flow. Rat tail bleeding times were obtained, and blood samples were drawn to determine fibrinogen and plasma protein concentrations, Lee-White clotting times, and activated partial thromboplastin times (APTT). Increased patency was found in the ancrod group (88%) (P < 0.05 vs. controls); turpentine-treated and control groups were not significantly different (71% and 63%, respectively). The ancrod group also showed significantly prolonged tail bleeding times and APTT. Fibrinogen levels were significantly decreased in the ancrod group (1.73 mg/ml) and elevated in the turpentine group (4.91 mg/ml) vs. controls (2.34 mg/ml; P < 0.005). These results indicate that elevated fibrinogen levels, in particular when triggered by an acute-phase response, do not appear to predispose small vessel repairs toward thrombosis. Furthermore, this study supports the use of ancrod as an anticoagulant for microvascular surgery.


Probing the p-type Chemiresistive Response of NiFe2 O4 Nanoparticles for Potential Utilization as Ethanol Sensor.
Detection of gas molecules and volatile organic compounds (VOCs) using efficient, low cost sensors has fetched significant attention in environmental monitoring, safety measures and medical diagnosis. In the present work, nickel ferrite (NFO) nanoparticles are explored as p-type semiconducting metal oxide (SMO) sensor for detection of five different organic vapors namely methanol, ethanol, n-propanol, iso-propanol and acetone which often cause severe damage to human body under prolonged exposure. The sensing studies in presence of the aforementioned five vapors are carried out by varying the sensor operating temperature (225-300 °C) and vapor concentrations (10-1000 ppm). Developed NFO sensor demonstrated best performance in terms of sensing (~10 ppm), response time (<10 s), excellent repeatability and selectivity towards ethanol among all other considered gas species. The repeatability of the sensor response is verified and the underlying reasons for the variation in the response of NFO sensor due to the change of operating temperature, analyte type and concentrations has been discussed. The synthesis of NFO through auto combustion method and study on their formation behaviour, oxygen vacancy evolution, band gap calculation, crystalline nature as well as microstructural features provides here the comprehensive information about the potential application of NFO nanoparticles as gas sensor.


Spinal holocord epidural abscess evacuated with double thoracic interval laminectomy: a rare case report with literature review.
Holocord spinal cord epidural abscess is an uncommon condition that may result in serious neurological complications. Prompt diagnosis and early treatment is of paramount importance for an optimum clinical outcome. This case report describes a novel technique of interval laminectomy at two sites in the thoracic spine and surgical decompression with the help of infant feeding tubes in a case of holocord spinal epidural abscess (HSEA). An 18-year-old male presented to the emergency department with high-grade fever and low back ache of 2 weeks duration and loss of bowel and bladder control for 4 days. Neurological examination revealed intact motor power and sensation in all four limbs at presentation; however, there was a rapid deterioration to complete quadriplegia within 24 h. A diagnosis of holocord epidural abscess was made. Emergent decompression via interval thoracic laminectomy was done and appropriate antimicrobial therapy was instituted. At 10 months of follow-up, the individual showed complete neurological recovery. The technique used in this case is unique with respect to the level of laminectomy and the manoeuvre employed for pus evacuation. Complete neurological and functional recovery was achieved despite complete paralysis pre-operatively. The outcome indicates that there may be good prognosis for individuals with HSEA accompanied with neurological deficit and emergent surgical decompression.


Long-term complications of implantable cardioverter defibrillator lead systems.
Over a period of 8.5 years in 255 patients with full-system implantable cardioverter defibrillators, lead-specific complications requiring reoperation developed in 32 patients. A total of 36 leads were affected. Lead fracture was the cause of failure in half of these patients. Refinement and improvement in structural designs of these leads, use of endocardial leads, and attention to technical details during implantation should decrease the incidence of these complications.


Inductive biases in theory-based reinforcement learning.
Understanding the inductive biases that allow humans to learn in complex environments has been an important goal of cognitive science. Yet, while we have discovered much about human biases in specific learning domains, much of this research has focused on simple tasks that lack the complexity of the real world. In contrast, video games involving agents and objects embedded in richly structured systems provide an experimentally tractable proxy for real-world complexity. Recent work has suggested that key aspects of human learning in domains like video games can be captured by model-based reinforcement learning (RL) with object-oriented relational models-what we term theory-based RL. Restricting the model class in this way provides an inductive bias that dramatically increases learning efficiency, but in this paper we show that humans employ a stronger set of biases in addition to syntactic constraints on the structure of theories. In particular, we catalog a set of semantic biases that constrain the content of theories. Building these semantic biases into a theory-based RL system produces more human-like learning in video game environments.


Itch and scratch: scabies and pediculosis.
Scabies and lice infestations are almost an everyday diagnosis in the dermatology clinics. Along with the unbearable itch, resistance to medication, secondary infection, and the high risk for spreading the parasite to their close contacts, the patients have also to battle with many myths, prejudice, and shames connected to those infestations.


Dynamic theta/beta ratio of clinical EEG in Alzheimer's disease.
EEG of a resting state in Alzheimer disease (AD) patients and healthy controls (HC) are analyzed to identify the characteristics of EEG in AD. A dynamic box plot approach to the theta/beta ratio with various window durations is proposed to analyze EEG. Spectral results during a resting state in AD patients demonstrate the effect of relatively greater power in the low-frequency bands (i.e. 'slowing down' of the EEG). A significant difference is observed in the dynamic distribution of the theta/beta ratio in the AD and HC groups, which is related to the effect of 'slowing down'. There is a more obvious visual separation between the theta/beta ratio results for the AD and HC groups with increasing window durations. Variability of the theta/beta ratio can be observed with shorter window durations with a dynamic functional box plot. This provides a better classification accuracy by using the dynamic theta/beta ratio as a sensor to discriminate AD EEG from HC EEG by using the receiver operating characteristics (ROC) curve and the area under curve (AUC) with various window durations. EEG spectral analysis and theta/beta ratio used to evaluate EEG typically rely on long time averaging. The dynamic box plot approach to the theta/beta ratio with various window durations provides the possibility of observing features of the EEG. The dynamic theta/beta ratio is a better sensor to discriminate AD EEG from HC EEG. Moreover, the reliability and accuracy of results can be increased by combining spectral analysis and the dynamic box plot approach to theta/beta ratio with various window durations.


Validation of bovine hoof slices as a model for infected human toenails: in vitro ciclopirox transungual permeation.
Topical therapy has recently been proposed for treating onychomycosis and other nail disturbances. However, the clinical outcome may be limited by the difficulty of active ingredients effectively penetrating the nail plate. Bovine hoof membranes have been widely used to predict in vitro efficacy of drug products in nail diseases. Many studies have compared bovine hooves with human healthy nails, considering the difference between healthy and unhealthy nails to be negligible. To validate bovine hoof slices as a model for human unhealthy nails by investigating the transungual permeation/retention of ciclopirox (CPX) through bovine hoof slices and excised infected human toenails after application of a new film-forming formulation (P-3051). To investigate the ability of CPX to achieve fungicidal concentrations in and through infected toenails. A new experimental technique based on a permeation unit allowed analysis by high-performance liquid chromatography of the amounts of CPX permeating through and retained in the membranes. The efficacy index was evaluated as follows: amount of permeated CPX/Trichophyton rubrum minimum inhibiting concentration. Extrapolated CPX flux through bovine hoof slices was about 14-fold higher than through infected human toenails, the difference being mainly due to the fourfold higher thickness of the toenails. In toenails, the CPX efficacy index for T. rubrum was positive (>1·0) soon after P-3051 application. This study confirms the validity of bovine hoof slices as a model for infected human nails, and suggests a substantial equivalence between the two models. Following P-3051 application, CPX reaches fungicidal concentrations in and through human infected toenails.


CRISPRi-mediated depletion of Spt4 and Spt5 reveals a role for DSIF in the control of HIV latency.
Pivotal studies on the control of HIV transcription has laid the foundations for our understanding of how metazoan transcription is executed, and what are the factors that control this step. Part of this work established a role for DRB Sensitivity Inducing Factor (DSIF), consisting of Spt4 and Spt5, in promoting pause-release of RNA Polymerase II (Pol II) for optimal elongation. However, while there has been substantial progress in understanding the role of DSIF in mediating HIV gene transcription, its involvement in establishing viral latency has not been explored. Moreover, the effects of depleting Spt4 or Spt5, or simultaneously knocking down both subunits of DSIF have not been examined. In this study, we employed CRISPR interference (CRIPSRi) to knockdown the expression of Spt4, Spt5 or the entire DSIF complex, and monitored effects on HIV transcription and viral latency. Knocking down DSIF, or each of its subunits, inhibited HIV transcription, primarily at the step of Tat transactivation. This was accompanied by a decrease in promoter occupancy of Pol II and Cdk9, and to a lesser extent, AFF4. Interestingly, targeting the expression of one subunit of DSIF, reduced the protein stability of its counterpart partner. Moreover, depletion of Spt4, Spt5 or DSIF complex impaired cell growth, but did not cause cell death. Finally, knockdown of Spt4, Spt5 or DSIF, facilitated entry of HIV into latency. We conclude that each DSIF subunit plays a role in maintaining the stability of its other partner, achieving optimal function of the DSIF to enhance viral gene transcription.


Differences in the location of bleeding with direct oral anticoagulants vs. vitamin K antagonists: A study in the World Health Organization's pharmacovigilance database.
Clinical trials have found differences in bleeding locations between direct oral anticoagulants (DOAC) and vitamin K antagonists (VKA). The present study was performed to investigate these differences in real life using reports of adverse drug reactions registered in the World Health Organization's pharmacovigilance database, VigiBase®. All bleeding registered between 1 January 2008 and 31 December 2021 in adults were included. The main objective was to compare bleeding locations reported with DOAC with those with VKA. As a secondary objective, we performed the same comparison with Xa vs. thrombin inhibitors. Results were presented as reporting odds ratios (RORs) adjusted on age, gender, origin of reports and co-medications with their 95% confidence interval. During this 14-year period, 142 228 instances of bleeding were registered with oral anticoagulants, including 39 570 with VKA and 102 658 with DOAC. Mean time to event was lower with DOAC (7.6 months) than with VKA (29.9 months) (P < .001). Significant differences in bleeding locations were found in the reports with less cerebral, urologic and nasal bleeding, more gynaecologic bleeding with DOAC than with VKA, without any significant differences in digestive and cutaneous locations. A higher risk of bleeding reports was found with Xa inhibitors vs. dabigatran whatever the locations (except digestive bleeding). This real-life study shows that the differences in bleeding locations between DOAC and VKA are not limited to the brain or gastrointestinal tracts. Significant differences were also found between Xa and thrombin inhibitors.


2,3-butanediol production by acetogenic bacteria, an alternative route to chemical synthesis, using industrial waste gas.
2,3-Butanediol (23BD) is a high-value chemical usually produced petrochemically but which can also be synthesized by some bacteria. To date, the best microbial 23BD production rates have been observed using pathogenic bacteria in fermentation systems that depend on sugars as the carbon and energy sources for product synthesis. Here we present evidence of 23BD production by three nonpathogenic acetogenic Clostridium species-Clostridium autoethanogenum, C. ljungdahlii, and C. ragsdalei-using carbon monoxide-containing industrial waste gases or syngas as the sole source of carbon and energy. Through an analysis of the C. ljungdahlii genome, the complete pathway from carbon monoxide to 23BD has been proposed. Homologues of the genes involved in this pathway were also confirmed for the other two species investigated. A gene expression study demonstrates a correlation between mRNA accumulation from 23BD biosynthetic genes and the onset of 23BD production, while a broader expression study of Wood-Ljungdahl pathway genes provides a transcription-level view of one of the oldest existing biochemical pathways.


Nosocomial endophthalmitis survey. Current incidence of infection after intraocular surgery.
The authors reviewed the incidence of hospital-linked postoperative endophthalmitis at the Bascom Palmer Eye Institute between January 1, 1984 and June 30, 1989. After 30,002 intraocular surgical procedures, the following incidence of culture-proven endophthalmitis was observed: (1) extracapsular cataract extraction (ECCE) with or without intraocular lens (IOL) implantation--0.072% (17 of 23,625 cases); (2) pars plana vitrectomy--0.051% (1 of 1974 cases); (3) penetrating keratoplasty (PKP)--0.11% (2 of 1783 cases); (4) secondary IOL--0.30% (3 of 988 cases); and (5) glaucoma filtering surgery--0.061% (1 of 1632 cases). A statistically significant (P = 0.038, Fisher's exact test, two-tailed) increased incidence of endophthalmitis occurred in diabetic (0.163%, 6 of 3686 cases) compared with nondiabetic (0.055%, 11 of 19,939 cases) patients undergoing ECCE with or without IOL implantation. The authors also reviewed the incidence of postoperative endophthalmitis after intracapsular cataract extraction (ICCE) with and without IOL and observed an incidence of 0.093% (7 of 7552) in cases operated on between September 1, 1976 and December 31, 1982.


Tryptophan transport through transport system T in the human erythrocyte, the Ehrlich cell and the rat intestine.
We studied the transport of tryptophan through transport system T in the human red cell, the Ehrlich ascites-tumour cell and in everted sacs of rat intestine. In red cells we confirmed earlier results on Na+-independence and aromatic amino acid specificity (Rosenberg, R., Young, J.D. and Ellory, J.C. (1980) Biochim. Biophys. Acta 598, 375-384). In addition we observed that N-methylation or N-acetylation did not reduce the affinity of the substrates for system T, hydroxylation could increase or decrease substrate affinity, and system T was insensitive to pH changes in the medium. These results characterized reactive differences between system T and other known amino acid transport systems. We also found that D-isomers were about 1/3 as effective as L-isomers to inhibit L-tryptophan uptake. D-Tryptophan competitively inhibited L-tryptophan uptake, but was not taken up by system T. L-Tryptophan produced trans-stimulation of the uptake (influx) and trans-inhibition of the release (efflux) of L-[3H]tryptophan; D-tryptophan produced trans-inhibition of the efflux but did not affect significantly the uptake. These results show that in red cells the transport properties of transport system T are asymmetric. Transport system T seems to be absent in the other two preparations studied, the Ehrlich ascites-tumour cell and the rat intestine.


New legislation to improve Canada's refugee system troubles advocates.
Despite significant improvements, the new Balanced Refugee Reform Act (the Act) contains contentious provisions regarding Canada's asylum system, according to refugee advocates.


What influences physician opioid prescribing for children with acute pain?
Pain is one of the most common symptoms encountered in the healthcare system, and opioids are among the top three medications used to treat it. Understanding the reasoning behind physicians' opioid prescribing practices is vital to safe practice. The primary objective of our study was to describe pediatric emergency physicians' decision-making process when prescribing opioids for children's acute pain management. This study employed qualitative methodology, using one-on-one semi-structured interviews within a grounded theory analytic framework. We employed purposeful sampling to recruit pediatric emergency physicians from across Canada. Interviews were conducted by telephone (December 2019-January 2021). Transcript analysis occurred concurrently with data collection, supporting data saturation and theory development considerations. Eleven interviews were completed with participants representing each of Canada's geographic regions. Nine major themes emerged: (1) practice setting and outpatient opioid use, (2) condition-specific considerations, (3) physician confidence in medical evidence, (4) pain assessment challenges, (5) patient and family perspectives, (6) opioid safety concerns, (7) personal biases and experiences, (8) personal practice context, and (9) the Opioid Crisis/media influence. Most clinicians felt that they limited opioid use to those who needed it most; all participants described challenges managing acute pain, emphasizing the need for accurate pain measurement and better guidelines, evidence-based data, and knowledge translation. Clinicians were more comfortable treating pain in the emergency department, compared to discharge prescribing. They recognized the importance of co-therapy with non-opioids and the need for opioid risk assessment when prescribing. A family centered approach was recognized as the goal of practice. Clinicians are less comfortable prescribing opioids to children for at-home use and find pain assessment and lack of clear guidelines to be barriers to pain care. Knowledge translation strategies for safer practice and optimal acute pain management could support responsible and judicious opioid use.


Relative contribution of walking velocity and stepping frequency to the neural control of locomotion.
Stepping frequency is tightly coupled to walking velocity during natural locomotion. In a recent model, we demonstrated that walking velocity determines stride frequency, governs the active feedback control of the swing and determines the swing phase dynamics that governs foot movement. Here, we questioned whether the swing phase dynamics reflect independent effects of stride frequency and walking velocity. Foot movements were measured with a motion detection system (Optotrak) while subjects walked at 0.6-2.1 m/s on a treadmill. Stepping frequencies of 1.3-2.8 Hz were generated with pacing cues at each walking velocity. In the 'iso-velocity' condition, peak forward toe velocity during the swing phases was related to walking velocity and did not vary with alterations in stride frequency. In the 'iso-frequency' condition, in contrast, stepping frequency altered the relationship between toe acceleration and toe position in the fore-aft direction. The cycle frequency, main sequence (peak velocity vs. amplitude) relationships, and the shape of the phase-plane trajectories of the swing phases also reflected this relationship. The data were modeled by decoupling stepping frequency from walking velocity, while maintaining active feedback control dependent on frequency. The latter predicted both the dominant shape of the phase plane trajectories and the main sequence relationships. Thus, according to the model, walking velocity and stride frequency are independent central variables that control the dynamics of the swing phases and stepping. The ability to decouple stride frequency from walking velocity may help in navigating over uneven terrain or when executing curved trajectories while maintaining a constant velocity.


Bilateral Pedicled Scrotal Flaps as an Alternative to Skin Graft in Penile Shaft Defects Repair.
To analyze surgical and functional outcomes of bilateral pedicled scrotal flaps for penile shaft reconstruction. A retrospective analysis was performed on 22 patients who underwent penile shaft reconstruction with bilateral pedicled scrotal flaps between 2009 and 2017. Demographics, peri-operative data, and surgical complications were collected. Functional outcomes were analyzed using a questionnaire made of the erection hardness score, the patient and observer scar assessment scale, and a 10-point Likert scale measuring patients... satisfaction about their skin coloration, sensitivity, elasticity and thickness, penile size, scrotal volume, erection quality, penetration ability, pain, sexual satisfaction, body image, masculinity, self-esteem, and global satisfaction. Patients exhibited a wide range of indications, including buried penis (27.2%), or subcutaneous injections of foreign material (27.2%). Early complications were suture dehiscence (31.8%), infection (13.6%) and hematoma (4.6%), associated with 9.1% of surgical revisions. Late complications were skin retraction (27.3%), testicular ascension (22.7%), pyramidal shape (4.6%) or shortening (13.6%) of the penis, associated with 27.3% of surgical revisions. For the 12 patients who answered the questionnaire, median erection hardness score and patient and observer scar assessment scale score [IQR] were 3.5 out of 4 [2.5-4] and 11.5 out of 60 [9.5-22], respectively. The patients reported a positive impact of the surgery on their psychological condition, with a median score of global satisfaction of 8 [IQR 7.5-9.5]. Bilateral pedicled scrotal flaps seem.ßto be a safe alternative for shaft defects reconstruction despite a potential need of surgical revision, providing satisfactory functional outcomes.


Olanzapine augmentation of paroxetine-refractory obsessive-compulsive disorder.
The aim of the present study was to investigate the effect of adjunctive olanzapine in patients with obsessive-compulsive disorder (OCD) refractory to paroxetine. Twenty-one patients unresponsive to treatment with paroxetine, administered for at least 12 weeks at the dose of 60 mg/day, participated to a 12-week open-label, add-on trial with olanzapine (10 mg/day). The psychopathological state was evaluated by the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) and by Clinical Global Impression (CGI). Three patients did not complete the 12-week adjunctive treatment with olanzapine. In the 18 completers, the mean Y-BOCS score decreased significantly from 27.1+/-4.0 at baseline to 20.1+/-3.9 at final evaluation (P<.001). Seven patients (38.9%) were rated as responders at final evaluation. Steady-state plasma concentrations of paroxetine were not modified during olanzapine coadministration. The drug combination was generally well tolerated and initial sedation and weight gain were the most frequent unwanted effects. Our findings confirm the results of previous studies and indicate that the addition of olanzapine to ongoing treatment with serotonin reuptake inhibitors (SRI) may be beneficial in some patients unresponsive to SRI monotherapy.


Higher risk of cytomegalovirus reactivation in human immunodeficiency virus-1-infected patients homozygous for MICA5.1.
Infection with cytomegalovirus (CMV) induces surface expression of major histocompatibility complex (MHC)-class-I-chain-related A (MICA), a ligand for NKG2D. This leads to improved recognition and elimination of infected cells by natural killer (NK) as well as CD8+ T cells. The MICA5.1 allele codes for a truncated protein. This study was performed to test whether impaired expression of a functional MICA protein would influence the susceptibility to severe CMV reactivation in immunocompromised individuals. In this study, the frequency of MICA5.1 was assessed by polymerase chain reaction in 230 Caucasian human immunodeficiency virus (HIV)-1-infected patients and in 219 healthy controls. Patients co-infected with hepatitis C virus (HCV) and GB virus-C served as controls. MICA5.1 allele was analyzed by polymerase chain reaction. Association of MICA5.1 homozygosity and risk of CMV reactivation was calculated by Pearson chi2 test. Comparison of patients with and without a history of CMV disease manifestation revealed that homozygous MICA5.1 genotype was present in a significantly higher frequency in patients with CMV reactivation (33%) than in those without (16%; p 0.032; odds ratio 0.330). The percentage was similar in HIV-1-infected patients and healthy controls. Furthermore, there was no difference in the frequency of MICA5.1 with respect to infection with HCV and GB virus-C. Our study provides the first in vivo demonstration of an association between homozygous MICA5.1 genotype and susceptibility to CMV reactivation in immunocompromised individuals.


Women's successful transition to motherhood during the early postnatal period: A qualitative systematic review of postnatal and midwifery home care literature.
To undertake a systematic review of available qualitative research literature to determine what women need to successfully transition to motherhood during the early postnatal period and whether postnatal home care delivered by midwives supports this process. The transition from woman to mother is a significant occurrence, one that requires adjustments on physical, psychological and social levels. A qualitative systematic review design was chosen to ensure both humanistic and interactive concepts related to what women need to successfully transition to motherhood during the early postnatal period were identified. The systematic and structured search uncovered 33 research articles for detailed review. Two team members, using the qualitative research checklist from the Critical Appraisal Skills Programme, appraised the quality of the research articles. A total of 19 articles met the quality criteria and were included in the data evaluation process. Research teams from Asia-Pacific, Europe, Middle East and North America generated the 19 qualitative journal articles. Data evaluation included identification of research aim, presence of a research question, type of methodology, data collection processes, sample information, data analysis techniques and study outcomes. Data was analysed using an inductive content analysis approach. Four themes were identified from the qualitative systematic review process including: women and midwives connecting, identification and meeting of women's individual needs, family and cultural influences and, education and support. The ability of women to connect with midwives during the early postnatal period assisted them overcome barriers and to successfully transitioning to motherhood. Although all four themes were determined to be key to effective postnatal transition, overwhelmingly the findings showed postnatal midwifery home care to be important in women's successful transition to motherhood in the early postnatal period. Strategies are needed ensure women have access to midwives in the early postnatal period.


Estimating hourly PM2.5 concentrations in Beijing with satellite aerosol optical depth and a random forest approach.
Assessing short-term exposure to PM2.5 requires the concentration distribution at a high spatiotemporal resolution. Abundant researches have derived the daily predictions of fine particles, but estimating hourly PM2.5 is still a challenge restrained by the input data. The recent aerosol optical depth (AOD) product from Himawari-8 provides hourly satellite observations informative to modelling. In this study, we developed separate random forest models with and without AOD and combined the estimates to obtain a full-coverage hourly PM2.5 distribution. 10-fold cross validation R2 ranged from 0.92 to 0.95 and root mean square errors from 14.1 to 16.9 μg/m3, indicating the good model performance. Spatial convolutional layers of PM2.5 measurements and temporal accumulation effects of meteorological features were added into the model. They turned out to be of the most important predictors and improved the performance significantly. Finally, we mapped hourly PM2.5 at a 1-km resolution in Beijing during a pollution episode in 2019 and studied the pollution pattern. The study proposed a method to obtain 24-h full-coverage hourly PM2.5 estimates which are useful for acute exposure assessment in epidemiological researches.


Enhancement of spontaneous acetylcholine release from motor nerve terminal by calmodulin inhibitors.
The role of calmodulin (CaM) in neuromuscular transmission in the rat diaphragm was electrophysiologically investigated using a microelectrode technique with two different CaM inhibitors, trifluoperazine (TFP) and N-(6-aminohexyl)-5-chrolo-1-naphthalene-sulfonamide HC1 (W-7). These inhibitors in the perfusate decreased the amplitude of miniature endplate potentials and increased the frequency dose dependently without any changes occurring in the resting membrane potentials. These effects were abolished in a Ca2+-free perfusate. The acetylcholine (ACh) quantal size and content were not affected by the reagents. The effects of TFP and W-7 were thought to result from their specific inhibition of calmodulin. It is suggested that accumulated intracellular Ca2+ and cyclic AMP, due to inhibition of calmodulin, had enhanced the frequency of spontaneous ACh release from the nerve terminal, and the decrease in the amplitude might be attributed to inhibition of the postsynaptic action of CaM.


Subcellular location and expression level of a chimeric protein consisting of the maize waxy transit peptide and the beta-glucuronidase of Escherichia coli in transgenic potato plants.
The transit peptide of the maize waxy protein (a nuclear-encoded amyloplast protein of the maize endosperm) was studied with respect to its role in subcellular protein targeting in transgenic potato plants. TP30, a chimeric precursor protein consisting of the waxy transit peptide and an additional 34 amino acids of the mature waxy protein fused to the beta-glucuronidase of Escherichia coli, was expressed in potato plants under the control of the 35S promoter of cauliflower mosaic virus. This fusion protein is imported not only into amyloplasts, the natural target organelles in the maize plant, but also into chloroplasts. In contrast, Gus, the beta-glucuronidase alone, which was also expressed in parallel experiments in transgenic potato plants is always found in the cytosol of the plant cells. As a consequence of the different subcellular locations of TP30 and Gus, we observed differences in the expression rates of the respective proteins in leaf cells, resulting in higher steady state levels of TP30 compared to Gus. In tuber cells, no correlation between intracellular location and expression of the proteins was found.


Aqua-(sulfato-κO)bis-[2-(1,3-thia-zol-4-yl-κN)-1H-benzimidazole-κN]iron(II).
In the title compound, [Fe(SO(4))(C(10)H(7)N(3)S)(2)(H(2)O)], the Fe(II) cation is sixfold coordinated by four N atoms from two 2-(1,3-thia-zol-4-yl)-1H-benzimidazole ligands, one water O atom and one O atom of the sulfate dianion within a slightly distorted octa-hedral geometry. The cations and anions are connected by N-H⋯O and O-H⋯O hydrogen bonds into layers in the ab plane.


Nitric oxide production by human monocytes: evidence for a role of CD23.
Nitric oxide (NO) appears to be an important and pleiotropic bioregulator of immune responses. The existence of the NO synthase (NOS) pathway in human monocytes/macrophages remains a subject of controversy, despite an increasing number of reports suggesting that human monocytes produce NO in vitro in response to various stimuli. Here, Bernard Dugas and colleagues consider the arguments supporting these conclusions, with particular emphasis on the results obtained by ligation of the low-affinity IgE receptor (Fcepsilon RIIb/CD23b).


Effectiveness of Remote Delayed Performance Feedback on Accurate Implementation of Caregiver Coaching.
The supervision of field experiences is an indispensable component of Board-Certified Behavior Analyst (BCBA®) training. During the supervised field experience, supervisors regularly provide performance feedback to trainees for the purpose of improving fidelity of implementation of various assessments and interventions. Emerging evidence supports the efficacy of using telehealth to train teachers and parents to implement interventions, but no study has evaluated the effectiveness of the remote delayed performance feedback among individuals completing BCBA® training. We used videoconference equipment and software to deliver remote delayed performance feedback to seven participants enrolled in a graduate program and completing supervised field experience. Remote delayed performance feedback was provided regarding participants' implementation of caregiver coaching. The results indicate that delayed performance feedback provided remotely increased the correct implementation of caregiver coaching. These preliminary results indicate the efficacy of remote supervision and delayed performance feedback.


Localized ultrasound enhances delivery of rapamycin from microbubbles to prevent smooth muscle proliferation.
Microbubble contrast agents have been shown to enhance reagent delivery when activated by ultrasound. We hypothesized that ultrasound would enhance delivery of rapamycin, an antiproliferative agent, from the shell of microbubbles, thus reducing proliferation of vascular smooth muscle cells. Our objective was to determine optimal ultrasound parameters that maximized therapeutic efficacy, maintained cell adherence, and minimized the drug exposure time. In vitro assays determined that ultrasound (1 MHz, 0.5% duty cycle) is required to successfully deliver rapamycin from microbubbles and reduce proliferation. Co-injection of rapamycin with control microbubbles did not result in a reduction in proliferation. Successful reduction in proliferation (>50%) required pulses at least 10 cycles in length and at least 300 kPa peak negative pressure at which point 90% of cells remained adherent. The anti-proliferative effect was also localized within a 6mm wide zone by focusing the ultrasound beam.


Perioperative therapeutic plasma exchange in a patient with rare Factor XIII inhibitor.
Factor XIII deficiency is a rare bleeding disorder which could be severe if inherited or less severe if acquired. We report a case of acquired Factor XIII inhibitor in a 75-year-old male with a suspicious left renal mass treated perioperatively with therapeutic plasma exchange (TPE). To perform kidney biopsy and ablation of the renal mass, six daily TPE treatments were performed before and after biopsy to minimize bleeding risk because the patient did not respond to drug therapy. Both thromboelastography (TEG) and laboratory-based coagulation tests were performed to assess coagulation status prior to and after TPE. The biopsy indicated oncocytoma which was removed by surgical procedure. Factor XIII activity remained below 15 % throughout TPE treatments, but Factor XIII inhibitor titer reduced from initial positive value of 1:40 to negative following the third TPE and remained negative through the sixth TPE. Unfortunately, the inhibitor titer was positive at 1:20 in the fifth month and 1:5 in the sixth month during follow-up. TPE is useful in removing XIII inhibitory factor, but the effects are only short term.


Non-associative phase separation in an evaporating droplet as a model for prebiotic compartmentalization.
The synthetic pathways of life's building blocks are envisaged to be through a series of complex prebiotic reactions and processes. However, the strategy to compartmentalize and concentrate biopolymers under prebiotic conditions remains elusive. Liquid-liquid phase separation is a mechanism by which membraneless organelles form inside cells, and has been hypothesized as a potential mechanism for prebiotic compartmentalization. Associative phase separation of oppositely charged species has been shown to partition RNA, but the strongly negative charge exhibited by RNA suggests that RNA-polycation interactions could inhibit RNA folding and its functioning inside the coacervates. Here, we present a prebiotically plausible pathway for non-associative phase separation within an evaporating all-aqueous sessile droplet. We quantitatively investigate the kinetic pathway of phase separation triggered by the non-uniform evaporation rate, together with the Marangoni flow-driven hydrodynamics inside the sessile droplet. With the ability to undergo liquid-liquid phase separation, the drying droplets provide a robust mechanism for formation of prebiotic membraneless compartments, as demonstrated by localization and storage of nucleic acids, in vitro transcription, as well as a three-fold enhancement of ribozyme activity. The compartmentalization mechanism illustrated in this model system is feasible on wet organophilic silica-rich surfaces during early molecular evolution.


Comparison of Biaxial Biomechanical Properties of Post-menopausal Human Prolapsed and Non-prolapsed Uterosacral Ligament.
Uterosacral ligaments (USLs) provide structural support to the female pelvic floor, and a loss of USL structural integrity or biomechanical function may induce pelvic organ prolapse (POP). Alterations in extracellular matrix composition and organization dictate USL mechanical function. Changes in USL microstructure and corresponding mechanical properties, however, are not fully understood, nor is it understood how microstructure and mechanics change with onset and progression of POP. This is due, in part, as USL properties are primarily characterized along a single direction (uniaxial test), whereas the USL is loaded in multiple directions simultaneously within the body. Biaxial testing permits the acquisition of biomechanical data from two axes simultaneously, and thus simulates a more physiologic assessment compared to the traditional uniaxial testing. Therefore, the objective of this study was to quantify the biaxial biomechanical properties and histological composition of the USL in post-menopausal women with and without POP at various stages. Potential correlations between tissue microstructural composition and mechanical function were also examined. Tangential modulus was lower and peak stretch higher in POP III/IV compared to non-POP and POP I/II in the main in vivo loading direction; however, no significant differences in mechanical properties were observed in the perpendicular loading direction. Collagen content positively correlated to tangential modulus in the main in vivo loading direction (r = 0.5, p = 0.02) and negatively correlated with the peak stretch in both the main in vivo (r = -0.5, p = 0.02) and perpendicular loading directions (r = -0.3, p = 0.05). However, no statistically significant differences in USL composition were observed, which may be due to the small sample size and high variability of small sections of human tissues. These results provide first step towards understanding what microstructural and mechanical changes may occur in the USL with POP onset and progression. Such information may provide important future insights into the development of new surgical reconstruction techniques and graft materials for POP treatment.


Three partial reactions of ribulose-bisphosphate carboxylase require both large and small subunits.
Three partial reactions of ribulose-bisphosphate carboxylase/oxygenase were measured in the presence and absence of small subunits using the enzyme from the cyanobacterium, Synechococcus ACMM 323, whose small subunits may be reversibly dissociated from its octameric, large-subunit core. These partial reactions were: the exchange of the proton at C-3 of the substrate, ribulose 1,5-bisphosphate, with the medium which is indicative of C-2, C-3 enolization; the hydrolysis of the 6-carbon reaction intermediate, 3-keto-2-carboxy-D-arabinitol 1,5-bisphosphate, to two molecules of 3-phosphoglycerate; and the decarboxylation of the 6-carbon intermediate, which is catalyzed only by the deactivated, divalent metal-ion-free carboxylase. None of these partial reactions was catalyzed by the small-subunit-depleted, large-subunit octamer to an extent greater than that expected from the residual small subunit content (about 3%), implying that small subunits are required for all three reactions. Clearly, the small subunit's influence is not restricted to any single stage of the catalytic sequence. Under conditions where it was possible to demonstrate tight binding of the reaction-intermediate analog, 2-carboxy-D-arabinitol 1,5-bisphosphate, to the large-subunit octamer, no binding of the 6-carbon intermediate could be detected. We suggest that either the tight-binding form of the 6-carbon intermediate is the hydrated gem-diol, not the ketone, or the large subunits by themselves intrinsically possess a trace of catalytic activity which discharges any bound intermediate before it can be measured.


Rates of molecular evolution and the fraction of nucleotide positions free to vary.
Selective constraints on DNA sequence change were incorporated into a model of DNA divergence by restricting substitutions to a subset of nucleotide positions. A simple model showed that both mutation rate and the fraction of nucleotide positions free to vary are strong determinants of DNA divergence over time. When divergence between two species approaches the fraction of positions free to vary, standard methods that correct for multiple mutations yield severe underestimates of the number of substitutions per site. A modified method appropriate for use with DNA sequence, restriction site, or thermal renaturation data is derived taking this fraction into account. The model also showed that the ratio of divergence in two gene classes (e.g., nuclear and mitochondrial) may vary widely over time even if the ratio of mutation rates remains constant. DNA sequence divergence data are used increasingly to detect differences in rates of molecular evolution. Often, variation in divergence rate is assumed to represent variation in mutation rate. The present model suggests that differing divergence rates among comparisons (either among gene classes or taxa) should be interpreted cautiously. Differences in the fraction of nucleotide positions free to vary can serve as an important alternative hypothesis to explain differences in DNA divergence rates.


Injuries after Hurricane Katrina among Gulf Coast Evacuees sheltered in Houston, Texas.
After Hurricane Katrina and a decline in the living conditions at a major temporary shelter in New Orleans, Louisiana, residents were offered transport to a Mega-Shelter in Houston, Texas. Approximately 200,000 Gulf Coast residents were transported to Houston's Astrodome/Reliant Center Complex for appropriate triage and transfer to other shelter facilities. The Katrina Clinic was quickly organized to treat evacuees with acute injuries and illnesses as well as chronic medical conditions. Clinic physicians documented 1130 hurricane-related injuries during Katrina Clinic's operational interval, September 1-22, 2005. This article documents the nature, extent, and location of injuries treated at that clinic. We compare the frequency of injury among Katrina evacuees who visited the clinic to that of injuries among clinic outpatient records recorded in a nationally representative database. Using the Barell Matrix system and codes from the International Classification of Diseases, Ninth Revision, we classify Katrina injuries by body region and nature of injury; we also document the large number of hurricane-related immunizations distributed at the temporary outpatient clinic. The results show a 42% higher injury proportion among Katrina evacuees and that approximately half of all of the evacuees required immunizations. Lower leg extremity injuries were among the most frequent injuries. Future planning for hurricanes should take into account nonfatal injuries requiring medical treatment and other supportive care.


Development of a new 32-plex InDels panel for forensic purpose.
Insertion/deletion polymorphisms (InDels) have attracted more and more attention of forensic researchers because of their low mutation rate and small amplicons. In the face of challenging forensic cases and degraded DNA, InDels break through the limitations of traditional STRs and provided a new direction for forensic identification. In this study, a multiplex panel consisting of 32 InDels and amelogenin was established and the InDels were selected with minimum allele frequencies (MAF) ≥ 0.4 in Chinese Han population based on the 1000 Genomes Project and published articles. The 32-plex InDels assay was evaluated using the southern Chinese Han population from Hubei province. All markers were proved to be highly polymorphic with an average observed heterozygosity (Hobs) of 0.4885. The combined power of discrimination (CPD) and the combined probability of paternity exclusion (CPE) were 0.999999999999966 and 0.9982, respectively. Further validation studies indicated that the 32-plex InDels assay was highly sensitive to DNA template amounts as low as 31.25 pg and was effective for degraded DNA. The obtained allele frequencies from this study were compared with published data of other populations.


Differential and common recognition of the catalytic sites of the cGMP-dependent and cAMP-dependent protein kinases by inhibitory peptides derived from the heat-stable inhibitor protein.
Synthetic peptides corresponding to the active domain of the heat-stable inhibitor protein of cAMP-dependent protein kinase (Cheng, H.-C., Kemp, B. E., Pearson, R. B., Smith, A. J., Misconi, L., Van Patten, S. M., and Walsh, D. A. (1986) J. Biol. Chem. 261, 989-992) were tested as inhibitors of cGMP-dependent protein kinase. The peptides themselves were not substrates. cGMP-dependent protein kinase activity was assayed using histone H2B and two synthetic peptide substrates. Consistent with previous observations of other peptide inhibitors of this enzyme (Glass, D. B. (1983) Biochem. J. 213, 159-164), the inhibitory peptides had no effect on the phosphorylation of histone H2B, but they competitively inhibited cGMP-dependent phosphorylation of the two peptide substrates. The parent inhibitor peptide, PKI(5-24)amide, and a series of analogs had Ki (or IC50) values for cGMP-dependent protein kinase in the range of 15-190 microM. In contrast to their effects on the cAMP-dependent protein kinase, the inhibitory peptides were substantially less potent with cGMP-dependent protein kinase, and potency was reduced by the presence of the NH2-terminal residues (residues 5-13). We conclude that the two protein kinases share a recognition of the basic amino acid cluster within the pseudosubstrate region of the peptide, but that the cGMP-dependent protein kinase does not recognize additional NH2-terminal determinants that make the inhibitor protein extremely potent toward the cAMP-dependent enzyme. Even- when tested at high concentrations and with peptide substrates, the native inhibitor protein did not inhibit cGMP-dependent protein kinase under assay conditions in which the peptides derived from it were inhibitory. Thus, the native inhibitor protein appears to have structural features which block interaction with the cGMP-dependent enzyme and enhance its selectivity for cAMP-dependent protein kinase.