Evaluation of FDG-PET and ECD-SPECT in patients with subcortical band heterotopia.
The purpose of this retrospective study was to clarify the cellular activities of ectopic neurons in subcortical bands and to evaluate the imaging features of (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and (99m)Tc ethyl cysteinate dimer (ECD) single-photon emission computed tomography (SPECT) in a series of patients with subcortical band heterotopia (SBH). The cases of 12 patients with SBH (3 men and 9 women; age range, 2-51 years) were evaluated on the basis of their MRI findings. Eight (18)F-FDG PET and 12 (99m)Tc-ECD SPECT images were obtained. The uptakes of these images were compared with electroencephalography (EEG) or MRI findings such as band thickness. In all patients, easy Z-score Imaging System (eZIS) software was used to statistically analyze the SPECT images. Of the eight (18)F-FDG PET images, five showed higher uptake in the thick subcortical bands than in the overlying cortex. Of the 12 (99m)Tc-ECD SPECT examinations with eZIS images, nine indicated increased regional cerebral blood flow (rCBF) areas corresponding to the band locations. Of the eight (18)F-FDG PET examination findings, six were congruent with the rCBF distributions on the eZIS images. Eight of the 12 patients showed correspondence to the increased rCBF on the eZIS images, the band locations on MRI, and abnormal discharge sites on EEG. Ectopic neurons in subcortical bands may have higher glucose metabolism and/or increased rCBF compared to the overlying cortex. (18)F-FDG PET and (99m)Tc-ECD SPECT using eZIS can be helpful to clearly detect the cellular activities of ectopic neurons in patients with SBH.


Determinants of early initiation of breastfeeding in The Gambia: a population-based study using the 2019-2020 demographic and health survey data.
Early initiation of breastfeeding within the first hour of life prevents neonatal and infant mortality. Sustainable Development Goals (SDGs) Target 3.2 aims to reduce neonatal mortality and under 5 mortality globally. The decline in the early initiation of breastfeeding in The Gambia coincides with deviations from the SDGs, due to poor indicators of child survival. Our work studied the determinants of early initiation of breastfeeding in The Gambia. We used the 2019-2020 Gambia Demographic Health Survey (GDHS) conducted across all regions of the country. Since our population of interest was children born two years preceding the study, we only included children less than 24 months of age, living with an eligible respondent. Thus, a weighted sample of 5691 mother-child pairs was applied in the analysis. We reported summary statistics of individuals' sociodemographic, obstetrics and antenatal, household, and community-level factors. A logistic regression model was used to determine associations between early initiation of breastfeeding and covariates. The prevalence of early initiation of breastfeeding was 64.3% (n = 3659). Mothers who had secondary education or higher educational level had higher odds of early initiation of breastfeeding (AOR 1.22; 95% CI 1.07, 1.40). Regions with rural population notably Lower and Central and Upper River Region had lower odds of early initiation of breastfeeding [Mansakonko (AOR 0.37; 95% CI 0.26, 0.15), Kerewan (AOR 0.26; 95% CI 0.19, 0.36), Kuntaur (AOR 0.39; 95% CI 0.28, 0.54), Janjanbureh (AOR 0.48; 95% CI 0.35, 0.66) and Basse (AOR 0.64; 95%CI 0.49, 0.85)]. Also, women in the high quintile of the wealth index were more likely to initiate breastfeeding early (AOR 1.29; 95% CI 1.06, 1.57). Four or more antenatal care visits did not increase early initiation of breastfeeding. The results of the analyses demand affirmative action to improve maternal education, reduce poverty and inequality and empower rural communities in The Gambia. The IYCF component in antenatal care needs to be strengthened. Programs and policies on IYCF must resonate to address determinants of timely breastfeeding initiation to chart progress towards the SDG.


Accounting carbon storage in decaying root systems of harvested forests.
Decaying root systems of harvested trees can be a significant component of belowground carbon storage, especially in intensively managed forests where harvest occurs repeatedly in relatively short rotations. Based on destructive sampling of root systems of harvested loblolly pine trees, we estimated that root systems contained about 32% (17.2 Mg ha(-1)) at the time of harvest, and about 13% (6.1 Mg ha(-1)) of the soil organic carbon 10 years later. Based on the published roundwood output data, we estimated belowground biomass at the time of harvest for loblolly-shortleaf pine forests harvested between 1995 and 2005 in South Carolina. We then calculated C that remained in the decomposing root systems in 2005 using the decay function developed for loblolly pine. Our calculations indicate that the amount of C stored in decaying roots of loblolly-shortleaf pine forests harvested between 1995 and 2005 in South Carolina was 7.1 Tg. Using a simple extrapolation method, we estimated 331.8 Tg C stored in the decomposing roots due to timber harvest from 1995 to 2005 in the conterminous USA. To fully account for the C stored in the decomposing roots of the US forests, future studies need (1) to quantify decay rates of coarse roots for major tree species in different regions, and (2) to develop a methodology that can determine C stock in decomposing roots resulting from natural mortality.


Effect of mold type, diameter, and uncured composite removal method on depth of cure.
This study compared the effects of mold material and diameter on the thickness of cured composite remnants and depth of cure (DOC) of resin-based composites (RBC). One Polywave® curing light was used to photo-cure two shades of the same "bulk-fill" RBC in 4, 6, or 10-mm internal diameter metal or white Delrin® molds. For 60 specimens, the uncured RBC was manually scraped away as described in the ISO 4049 depth of cure test. The remaining 60 specimens were immersed in tetrahydrofuran for 48 hours in the dark. Maximum lengths of remaining hard RBC and their DOC values were compared using analysis of variance (ANOVA) and Tukey-Kramer post hoc multiple comparison tests (α = 0.05). Specimen thickness and DOC were always greater using the white Delrin® molds compared to metal molds (p < 0.001). Increase in mold diameter significantly increased specimen thickness and DOC when made in the metal molds and in the 6-mm diameter Delrin® molds (p < 0.01). Increasing the diameter of the Delrin® molds to 10-mm did not increase specimen thickness or DOC. Sectioning and staining of specimens revealed an internal, peripheral transition zone of porous RBC in the solvent-dissolved specimens only. Mold material and internal diameter significantly influenced cured composite remnant thickness as well as depth of cure. The existence of an outer region of RBC that is hard, yet susceptible to solvent dissolution, requires further investigation. The depth of cure results obtained from a 4-mm diameter metal mold may not represent the true potential for evaluating composite depth of cure. A universally acceptable mold material and diameter size need to be established if this type of testing is to be useful for evaluating the relative performance of a given type of LCU and RBC.


Sensitive and rapid detection of blaNDM-1 in clinical samples by isothermal cross-priming amplification.
A highly sensitive, specific diagnostic assay for detection of bla(NDM-1) based on cross priming amplification (CPA) was developed. The sensitivity ranged from 2.5 to 25 copies per reaction for different clinical samples. The highly and sensitive detection of bla(NDM-1) in clinical samples highlighted the potential clinical applications of CPA.


Proliferation to paucity: evolution of bile duct abnormalities in a case of Alagille syndrome.
Alagille syndrome is an autosomal dominant disorder characterized by abnormalities in multiple organ systems, including the liver, and is caused by mutations in JAG1. Chronic cholestasis secondary to paucity of interlobular bile ducts is traditionally both a clinical and a pathologic hallmark of this disease at diagnosis. We describe the biliary changes on serial liver biopsies in a patient who presented with jaundice and extrahepatic stigmata of Alagille syndrome. Her initial specimens at 6 and 10 months of age demonstrated interlobular bile duct proliferation and cholestasis, suggestive of distal biliary obstruction. A specimen at 2 years of age showed near-total absence of interlobular bile ducts, with the classic histologic appearance of bile duct paucity. We present this case to underscore the potential pitfalls in interpreting cholestatic liver morphology in the absence of clinical information. The progression of bile duct abnormalities is discussed in the context of the role postulated for JAG1 in postnatal liver growth and development.


Use of Health Information Exchange and Repeat Imaging Costs.
The aim of this study was to quantify the association between health information exchange (HIE) use and cost savings attributable to repeat imaging. Imaging procedures associated with HIE were compared with concurrent controls on the basis of propensity score matching over the period from 2009 to 2010 in a longitudinal cohort study. The study sample (n = 12,620) included patients ages 18 years and older enrolled in the two largest commercial health plans in a 13-county region of western New York State served by the Rochester Regional Health Information Organization. The primary outcome was a continuous measure of costs associated with repeat imaging. The determinant of interest, HIE use, was defined as system access after the initial imaging procedure and before repeat imaging. HIE use was associated with an overall estimated annual savings of $32,460 in avoided repeat imaging, or $2.57 per patient. Basic imaging (radiography, ultrasound, and mammography) accounted for 85% of the estimated avoided cases of repeat imaging. Advanced imaging (CT and MRI) accounted for 13% of avoided procedures but constituted half of the estimated savings (50%). HIE systems may reduce costs associated with repeat imaging. Although inexpensive imaging procedures constituted the largest proportion of avoided repeat imaging in our study, most of the estimated cost savings were due to small reductions in repeated advanced imaging procedures. HIE systems will need to be leveraged in ways that facilitate greater reductions in advanced imaging to achieve appreciable cost savings.


The Dmp1-SOST Transgene Interacts With and Downregulates the Dmp1-Cre Transgene and the Rosa(Notch) Allele.
Activation of Notch1 in osteocytes of Rosa(Notch) mice, where a loxP-flanked STOP cassette and the Nicd coding sequence were targeted to the reverse orientation splice acceptor (Rosa)26 locus, causes osteopetrosis associated with suppressed Sost expression and enhanced Wnt signaling. To determine whether Sost downregulation mediates the effects of Notch activation in osteocytes, Rosa(Notch) mice were crossed with transgenics expressing Cre recombinase or SOST under the control of the dentin matrix protein (Dmp)1 promoter. Dmp1-SOST transgenics displayed vertebral osteopenia and a modest femoral cancellous and cortical bone phenotype, whereas hemizygous Dmp1-Cre transgenics heterozygous for the Rosa(Notch) allele (Dmp1-Cre;Rosa(Notch)) exhibited osteopetrosis. The phenotype of Notch activation in osteocytes was prevented in Dmp1-Cre;Rosa(Notch) mice hemizygous for the Dmp1-SOST transgene. The effect was associated with downregulated Notch signaling and suppressed Dmp1 and Rosa26 expression. To test whether SOST regulates Notch expression in osteocytes, cortical bone cultures from Dmp1-Cre;Rosa(Notch) mice or from Rosa(Notch) control littermates were exposed to recombinant human SOST. The addition of SOST had only modest effects on Notch target gene mRNA levels and suppressed Dmp1, but not Cre or Rosa26, expression. These findings suggest that prevention of the Dmp1-Cre;Rosa(Notch) skeletal phenotype by Dmp1-SOST is not secondary to SOST expression but to interactions among the Dmp1-SOST and Dmp1-Cre transgenes and the Rosa26 locus. In conclusion, the Dmp1-SOST transgene suppresses the expression of the Dmp1-Cre transgene and of Rosa26.


Dietary Vitamin C Deficiency Is Associated With Health-Related Quality of Life and Cardiac Event-free Survival in Adults With Heart Failure.
Vitamin C deficiency is prevalent in adults with heart failure (HF). Little is known about the relationship of dietary vitamin C deficiency with health outcomes in adults with HF. The study's aim was to determine the relationships of vitamin C deficiency measured at baseline with health-related quality of life (HRQOL) and cardiac event-free survival in patients with HF measured 1 year later. A total of 251 patients with HF completed a 4-day food diary. Dietary vitamin C deficiency was defined as daily intake less than the estimated average requirement from the Institute of Medicine of 75 mg/d for men and 60 mg/d for women. Health-related quality of life was assessed using the Minnesota Living with Heart Failure Questionnaire at 12 months. Patients were followed for a median of 1 year to determine time to the first event of cardiac-related hospitalization or death. Data were analyzed by hierarchical linear and Cox proportional hazards regressions. One hundred patients (40%) had vitamin C deficiency. Dietary vitamin C deficiency was associated with poorer HRQOL at 12 months (β = 0.16, P = .02) after controlling for demographic and clinical variables. During the follow-up period, 59 patients (24%) had cardiac events. In Cox regression, vitamin C deficiency predicted shorter cardiac event-free survival after adjusting for the same covariates (hazards ratio, 1.95; 95% confidence interval, 1.08-3.51). Vitamin C deficiency was associated with poorer HRQOL and shorter cardiac event-free survival in patients with HF. The findings suggest that encouraging patients with HF to consume a diet rich in fruits/vegetables to prevent vitamin C deficiency may lead to better health outcomes.


Hybrid Capture II detection of oncogenic human papillomavirus: a useful tool when evaluating men who have sex with men with atypical squamous cells of undetermined significance on anal cytology.
In the cervix and anus, patients with atypical squamous cells of undetermined significance often do not have high-grade squamous intraepithelial lesions. In women with atypical squamous cells of undetermined significance, Hybrid-Capture II testing for oncogenic high-risk human papillomavirus is performed and those without high-risk human papillomavirus often are observed. We endeavored to determine whether Hybrid-Capture II testing would be beneficial in men who have sex with men with atypical squamous cells of undetermined significance. We performed a retrospective chart review of men who have sex with men with atypical squamous cells of undetermined significance who had high-resolution anoscopy and Hybrid-Capture II. A total of 290 men were identified (mean age, 42 years), and 212 (73 percent) were HIV-negative. High-grade squamous intraepithelial lesions were found in 50 (17 percent): 23 (10 percent) who were HIV-negative and 27 (35 percent) who were HIV-positive men. High-risk human papillomavirus was found in 138 (48 percent); 91 (43 percent) of HIV-negative and 47 (60 percent) of HIV-positive men. The sensitivity, specificity, positive predictive value, and negative predictive value of atypical cells of undetermined significance cytology combined with Hybrid-Capture II were 84, 60, 30, and 95 percent, respectively. There was no significant difference between all men vs. those who were HIV-positive or HIV-negative except for the positive predictive value. Hybrid-Capture II testing for high-risk human papillomavirus in men who have sex with men with atypical cells of undetermined significance and referring only those with high-risk human papillomavirus reduces the number who require high-resolution anoscopy by more than half. Five percent with high-grade squamous intraepithelial lesions would be missed.


Complex life histories discovered in a critically endangered fish.
Effective conservation of endangered species requires knowledge of the full range of life-history strategies used to maximize population resilience within a stochastic and ever-changing environment. California's endemic Delta Smelt (Hypomesus transpacificus) is rapidly approaching extinction in the San Francisco Estuary, placing it in the crossfire between human and environmental uses of limited freshwater resources. Though managed as a semi-anadromous species, recent studies have challenged this lifecycle model for Delta Smelt, suggesting the species is an estuarine resident with several localized "hot-spots" of abundance. Using laser-ablation otolith strontium isotope microchemistry, we discovered three distinct life-history phenotypes including freshwater resident (FWR), brackish-water resident (BWR), and semi-anadromous (SA) fish. We further refined life-history phenotypes using an unsupervised algorithm and hierarchical clustering and found that in the last resilient year-class, the FWR (12%) and BWR (7%) comprised a small portion of the population, while the majority of fish were SA (81%). Furthermore, the semi-anadromous fish could be clustered into at least four additional life-history phenotypes that varied by natal origin, dispersal age and adult salinity history. These diverse life-history strategies should be incorporated into future conservation and management efforts aimed at preventing the extinction of Delta Smelt in the wild.


On-line coupling of supercritical fluid extraction with enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry for the determination of chiral pesticides in hemp seeds: A proof-of-principle study.
The present research can be considered as a proof-of-principle study focused on the determination of chiral pesticides using a supercritical fluid extraction instrument coupled on-line with an enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry. To the best of Authors' knowledge, this is the first description of an on-line approach for the extraction and determination of chiral pesticides. Metalaxyl, benalaxyl and dimethenamid were investigated in nine hemp seed samples belonging to four varieties of Cannabis sativa; only in one case a pesticide was found at levels above the method limit of quantification (LoQ), though within the EU maximum residue level value. The figures-of-merit determined were linearity, precision, limit of detection (LoD), and LoQ. Regression coefficients were between 0.9856 and 0.9973, the LoDs were in the 0.04-0.41 μg kg-1 range, the LoQs were in the 0.12-1.38 μg kg-1 range, while coefficients of variation were between 1 and 3% (10 μg kg-1 level).


Tobacco smoke exposure is associated with the metabolic syndrome in adolescents.
The metabolic syndrome predicts future coronary artery disease and type II diabetes and often emerges in childhood. Tobacco smoke potentially contributes to insulin resistance in this syndrome. This study evaluates the association of environmental tobacco smoke (ETS) exposure and active smoking with the prevalence of the metabolic syndrome in US adolescents. Data from 2273 subjects 12 to 19 years of age were examined from the National Health and Nutrition Examination Survey III (NHANES III, 1988 to 1994). Serum cotinine levels, presence of household smokers, and self-report of smoking were used to determine ETS exposure and active smoking. The metabolic syndrome was defined as having > or =3 criteria from the National Cholesterol Education Panel definition. Bivariate and multivariable analyses were conducted. Among adolescents, 5.6% met the criteria for metabolic syndrome, and prevalence increased with tobacco exposure: 1.2% for nonexposed, 5.4% for those exposed to ETS, and 8.7% for active smokers (P<0.001). In adolescents at risk for overweight and overweight adolescents (body mass index above the 85th percentile), a similar relationship was observed: 5.6% for nonexposed, 19.6% for those exposed to ETS, and 23.6% for active smokers (P=0.01). In multivariable logistic regression analyses among all adolescents, ETS exposure was independently associated with the metabolic syndrome (ETS exposure: odds ratio, 4.7, 95% CI, 1.7 to 12.9; active smoking: odds ratio, 6.1; 95% CI, 2.8 to 13.4). Considering that tobacco and obesity are the 2 leading causes of preventable death in the United States, these findings of a dose-response, cotinine-confirmed relationship between tobacco smoke and metabolic syndrome among adolescents may have profound implications for the future health of the public.


Genomic landscape of secretory carcinoma of the breast with axillary lymph node metastasis.
Secretory carcinoma of the breast (SCB) is a rare low-grade often triple-negative breast carcinoma. We aim to analyze the pathological and molecular features of 21 SCBs, especially the SCBs with axillary lymph node metastasis. The clinicopathological characteristics of 21 SCBs were reviewed. Breast biomarkers, Pan-TRK and ETV6 break, and ETV6-NTRK3 fusion were performed on all cases. Next-Generation Sequencing (NGS) was performed on two cases with lymph node metastasis. 21 SCBs consisted of 2 men and 19 women aged 5~73 years (median 43 years), with a mean 2.1 cm (range 0.5~3.5 cm) tumor size. 90.1% (19/21) cases had mixed microcystic, solid, tubular, and papillary patterns. Pan-TRK and S100 are positive in 95% (20/21) and 90% (19/21) of cases, respectively. Tumor markers ER, PR, and HER2 expressions were 62% (13/21), 33% (7/21), and 0% (0/21). All cases showed ETV6 (21/21) rearrangement and ETV6-NTRK3 (11/11) fusion. 57% (12/21) of the cases had a balanced translocation and 38% (8/21) with unbalanced signals of ETV6. There was no clinical difference between balanced and unbalanced translocations in histological morphology and other prognosis factors. Furthermore, one case (#4) had a duplication of the ETV6 gene and presented axillary lymph node metastasis. NGS analysis revealed simple genomes, low tumor mutation burden, stable microsatellite sites, and single nucleotide polymorphism (SNP) heterozygous mutation in both SCBs with nodal metastasis. SCB is an indolent invasive carcinoma, even the cases with axillary lymph node metastasis, presenting simple genomes. Duplication of ETV6 cases may indicate lymph node metastasis.


[Mortality caused by pleural mesothelioma in the 1979-1983 lustre in Italy regarding provinces and single local health districts].
The geographic distribution of deaths from pleural mesothelioma in Italian provinces was analysed over the five-year period 1979-1983. In the provinces where the mortality rates were markedly higher than the national average, a further analysis was made on the data referred to the local health districts (USSL) of each province. It was thus possible, in some cases, to identify small, high risk areas which corresponded, as expected, to the location of large factories or shipyards using asbestos.


Cadaveric sperm induces intergeneric androgenesis in the fish, Hemigrammus caudovittatus.
Intergeneric androgenetic golden Buenos Aires tetra (BT), Hemigrammus caudovittatus was generated using sperm drawn from post-mortem males preserved at -20 degrees C for 10, 20, 30 and 40 days or fresh sperm to activate the UV-irradiated oocytes of black widow tetra (WT), Gymnocorymbus ternetzi. UV-irradiation (4.2 W/m(2)) of the oocytes for 3 min inactivated their nuclear genome. Fry hatched out from these activated oocytes were haploids; suffering haploid syndrome, they died before or within 48 h after hatching. Fresh BT sperm activated 95% oocytes; however, the sperm drawn from post-mortem males preserved at -20 degrees C for 60 (within glycerol packing) and 30 days (without glycerol packing) activated only 24 and 19% oocytes, respectively. Following activation, diploidy was restored by shocking the 25-min-old embryos at 41 degrees C for 2 min. Nuclear genomic inactivation of the oocytes was confirmed by (i) production of 100% haploids, (ii) karyotype and erythrocyte measurements, (iii) phenotypic markers, (iv) progeny testing and (v) species-specific marker. At hatching, survival of androgenotes decreased from 11% for those induced with fresh sperm to 4% for those generated using sperm from 30-day-old post-mortem males. Reproductive performance of the 'fresh' and 'cadaveric' F(0) and F(1) androgenetic males (Y(2)Y(2)) was superior to the control (X(1)Y(2)). Crosses involving homozygous (Y(2)Y(2)) 'fresh' F(0) androgenetic males with heterozygous females (X(1)X(2)) and F(0) homozygous males (Y(2)Y(2)) with females (X(2)X(2)) produced 2-4% unexpected female progenies. Paternal autosomes, inherited by the homozygous androgenetic female (X(2)X(2)), induced the production of female progenies in significantly less number of crosses than the crosses with heterozygous females (X(1)X(2)), which carried equal number of paternal and maternal autosomes. PCR analyses of the genomic DNA of normal male and unexpected F(1) and F(2) female progenies amplified by DMRT 1 specific primer produced bands of 237 and 300 bp length, and thereby confirmed that these unexpected females were genetic males. RAPD analyses of the androgenetic progenies showed that their genome was not contaminated with maternal genome.


A prospective cohort study of shift work and the risk of death from pancreatic cancer in Japanese men.
There is mounting evidence that shift work involving night work increases cancer risk. We examined the relationship between working rotating shifts and the risk of death from pancreatic cancer on the basis of data from the Japanese Collaborative Cohort Study (JACC Study). The present analysis was restricted to 22,224 men who were 40-65 years of age at baseline (1988-1990) and who reported working full time or were self-employed in the JACC Study. The subjects were followed through 31 December 2009. Information on occupation and lifestyle factors was collected using a self-administered questionnaire. The Cox proportional hazards model was used to estimate the relative risk (RR) and 95% confidence interval (CI) for the risk of death from pancreatic cancer in relation to shift work. During the follow-up period, 127 pancreatic cancer deaths were observed. Overall, we found no statistically significant increase in the risk of death from pancreatic cancer associated with rotating shift work. As compared to day-shift workers, the RRs were 0.83 (95% CI 0.43-1.60) for rotating shift workers and 0.61 (95% CI 0.22-1.60) for fixed night-shift workers, after adjustment for potential confounding factors. The multivariable-adjusted RR was 1.34 (95% CI 0.66-2.75) among rotating shift workers in the analysis restricted to men who reported working full time at baseline. Our data did not support the hypothesis that shift work is significantly associated with the risk of death from pancreatic cancer in this cohort of Japanese men.


The impact of somatic mosaicism on bicuspid aortic valve and aortic dissection in Turner Syndrome.
Somatic mosaicism is a major modifier of turner syndrome (TS) features and may be more prevalent than once thought, as new molecular techniques enable detection of tissue-specific mosaicism. This review explores the causes of mosaicism, discusses how mosaicism may impact congenital aortic defects in TS, and summarizes molecular methods to detect mosaicism in different contexts.


Microencapsulation of CHO cells in a hydroxyethyl methacrylate-methyl methacrylate copolymer.
Chinese hamster ovary fibroblasts, as model cells, have been microencapsulated in a hydroxyethyl methacrylate-methyl methacrylate copolymer (HEMA-MMA) by interfacial precipitation. The polymer containing approximately equal to 75 mol% HEMA, dissolved in polyethylene glycol 200 (PEG 200) was coextruded with the cell suspension (4-6 X 10(5) cells/ml in the alpha-MEM with 10% foetal calf serum +/- Ficoll 400/PBS) through a concentric needle assembly. Polymer solution droplets, containing cells, were blown off the end of the needle assembly by a coaxial filtered air stream into a nonsolvent bath containing phosphate buffered saline (PBS) with 5 ppm Pluronic L101, overlaid with hexadecane. The nascent capsules hang at the hexadecane/PBS interface while the solvent is extracted into the aqueous nonsolvent, to precipitate the polymer around the cells. The resultant capsules were 500 microns-1 mm in diam. with a microporous sponge-like interior, and also very tough and flexible. The cells survived encapsulation based on subculture ability, retention of some fluorescein diacetate (FDA) activity over 5 d and direct light microscopic evidence of cell growth over 10 d after histological sectioning and staining. However, cell growth was not uniformly observed (especially in the FDA assay) and this was attributed to space limitations for growth within the microporous interior. Continued development of this process and adaptation to cells such as pancreatic islets is expected to lead to hybrid artificial organs which are capable of ameliorating metabolic disorders such as diabetes.


Is it time to screen women with history of hypertensive pregnancy disorders for diabetes?
Thach Tran discusses a new research article on whether preeclampsia is associated with future diabetes, and discusses whether women with a history of hypertensive disorders in pregnancy should be screened for diabetes.


Prognostic importance of glycemic variability on left ventricular reverse remodeling after the first episode of ST-segment elevation myocardial infarction.
This study aimed to investigate the effect of glycemic variability (GV), determined using a continuous glucose monitoring system (CGMS), on left ventricular reverse remodeling (LVRR) after ST-segment elevation myocardial infarction (STEMI). A total of 201 consecutive patients with STEMI who underwent reperfusion therapy within 12 h of onset were enrolled. GV was measured using a CGMS and determined as the mean amplitude of glycemic excursion (MAGE). Left ventricular volumetric parameters were measured using cardiac magnetic resonance imaging (CMRI). LVRR was defined as an absolute decrease in the LV end-systolic volume index of > 10% from 1 week to 7 months after admission. Associations were also examined between GV and LVRR and between LVRR and the incidence of major adverse cardiovascular events (MACE; cardiovascular death, acute coronary syndrome recurrence, non-fatal stroke, and heart failure hospitalization). The prevalence of LVRR was 28% (n = 57). The MAGE was independent predictor of LVRR (odds ratio [OR] 0.98, p = 0.002). Twenty patients experienced MACE during the follow-up period (median, 65 months). The incidence of MACE was lower in patients with LVRR than in those without (2% vs. 13%, p = 0.016). Low GV, determined using a CGMS, was significantly associated with LVRR, which might lead to a good prognosis. Further studies are needed to validate the importance of GV in LVRR in patients with STEMI.


Through-Space Electrostatic Effects of Positively Charged Substituents on the Hydrogen Evolution Reaction.
Elucidating the effects of various structural components on energy-related small molecule activation is of fundamental and practical significance. Herein the inhibition effect of positively charged substituents on the hydrogen evolution reaction (HER) was reported. With the use of Cu porphyrins 1-5 containing different numbers and locations of positively charged substituents, it was demonstrated that their electrocatalytic HER activities significantly decreased when more cationic units were located close to the Cu ion: the icat /ip (icat is the catalytic peak current, ip is the one-electron reduction peak current) value decreased from 38 with zero cationic unit to 15 with four closely located cationic units. Inspired by this result, Cu porphyrin 6, with four meso-phenyl groups each bearing a negatively charged para-sulfonic substituent, was designed. With these anionic units, 6 outperformed the other Cu porphyrins for electrocatalytic HER under the same conditions.


Metabolic products from microorganisms. 230. Amiclenomycin-peptides, new antimetabolites of biotin. Taxonomy, fermentation and biological properties.
Four new and two known peptide antibiotics containing amiclenomycin (Acm) have been isolated from a culture of Streptomyces venezuelae Tü 2460: L-MeIle-L-Acm (1), L-Ile-L-Acm (2), L-MeVal-L-Acm (3), L-MeIle-L-Acm-L-Gln (4), L-Ile-L-Acm-L-Gln (5) and L-Val-L-Acm-L-Gln (6). These di- and tripeptides exhibited antimicrobial activity on a minimal medium against Gram-negative bacteria, which could be reversed by biotin. It was shown that the antibiotics were decomposed by peptidases to provide amiclenomycin (7) after their uptake into cells of Escherichia coli via peptide-permeases. The antimicrobial activity of the amiclenomycin-peptides was the inhibition of DAPA-aminotransferase by the amiclenomycin-warhead, however, amiclenomycin itself was hardly transported into the cells. Since the amiclenomycin peptides misuse general transport systems, they are presented here as examples for the illicit transport concept.


Melanomas with concurrent BRAF non-p.V600 and NF1 loss-of-function mutations are targetable by BRAF/MEK inhibitor combination therapy.
Although combination BRAF/MEK inhibition has produced significant survival benefits for BRAF p.V600 mutant melanomas, targeted therapies approved for BRAF non-p.V600 mutant melanomas remain limited. Through the analysis of 772 cutaneous melanoma exomes, we reveal that BRAF non-p.V600 mutations co-occurs more frequently with NF1 loss, but not with oncogenic NRAS mutations, than expected by chance. We present cell signaling data, which demonstrate that BRAF non-p.V600 mutants can signal as monomers and dimers within an NF1 loss context. Concordantly, BRAF inhibitors that inhibit both monomeric and dimeric BRAF synergize with MEK inhibition to significantly reduce cell viability in vitro and tumor growth in vivo in BRAF non-p.V600 mutant melanomas with co-occurring NF1 loss-of-function mutations. Our data suggest that patients harboring BRAF non-p.V600 mutant melanomas may benefit from current FDA-approved BRAF/MEK inhibitor combination therapy currently reserved for BRAF p.V600 mutant patients.


Reducing the Risks of Firearm Violence in High Schools: Principals' Perceptions and Practices.
This study assessed the perceptions and practices of a national sample of secondary school principals regarding reducing firearm violence in high schools. Data were collected via three-wave postal mailings. A 59-item valid and reliable questionnaire was mailed to a national random sample of 800 secondary school principals. Of the 349 principals (46 %) that responded, 17 % reported a firearm incident at their school in the past 5 years. Principals perceived inadequate parental monitoring (70 %), inadequate mental health services (64 %), peer harassment/bullying (59 %), and easy access to firearms (50 %) as the main causes of firearm violence in schools. The three barriers to implementing firearm violence prevention practices were: lack of expertise as to which practices to implement (33 %), lack of time (30 %), and lack of research as to which practices are most effective (30 %). Less than half of schools trained school personnel regarding firearm violence issues. The findings indicate that firearm incidents at schools may be more common than previously thought. A significant portion of principals are at a loss as to what to implement because of a lack of empirical evidence on what is effective. More research is needed to find the most effective school interventions for reducing firearm violence.


Thromboprophylaxis in cancer outpatients.
In this issue of Blood, Khorana and colleagues report a simple and practical model for the prediction of symptomatic venous thrombosis after the initiation of chemotherapy in cancer outpatients.


Establishment and validation of a clinical model for predicting diabetic ketosis in patients with type 2 diabetes mellitus.
Diabetic ketosis (DK) is one of the leading causes of hospitalization among patients with diabetes. Failure to recognize DK symptoms may lead to complications, such as diabetic ketoacidosis, severe neurological morbidity, and death. This study aimed to develop and validate a model to predict DK in patients with type 2 diabetes mellitus (T2DM) based on both clinical and biochemical characteristics. A cross-sectional study was conducted by evaluating the records of 3,126 patients with T2DM, with or without DK, at The Affiliated Hospital of Qingdao University from January 2015 to May 2022. The patients were divided randomly into the model development (70%) or validation (30%) cohorts. A risk prediction model was constructed using a stepwise logistic regression analysis to assess the risk of DK in the model development cohort. This model was then validated using a second cohort of patients. The stepwise logistic regression analysis showed that the independent risk factors for DK in patients with T2DM were the 2-h postprandial C-peptide (2hCP) level, age, free fatty acids (FFA), and HbA1c. Based on these factors, we constructed a risk prediction model. The final risk prediction model was L= (0.472a - 0.202b - 0.078c + 0.005d - 4.299), where a = HbA1c level, b = 2hCP, c = age, and d = FFA. The area under the curve (AUC) was 0.917 (95% confidence interval [CI], 0.899-0.934; p<0.001). The discriminatory ability of the model was equivalent in the validation cohort (AUC, 0.922; 95% CI, 0.898-0.946; p<0.001). This study identified independent risk factors for DK in patients with T2DM and constructed a prediction model based on these factors. The present findings provide an easy-to-use, easily interpretable, and accessible clinical tool for predicting DK in patients with T2DM.


Response of patients with indolent systemic mastocytosis to tamoxifen citrate.
We examined whether tamoxifen citrate at 20mg/day for 1 year had a beneficial effect on laboratory findings, bone marrow mastocytosis, common clinical symptoms, or quality-of-life assessment for 5 women and 2 men with indolent systemic mastocytosis. Tamoxifen was well tolerated. We found significant reductions in the platelet count, serum alkaline phosphatase, and 24-h urinary excretion of N-methylhistamine and significant increases in serum lactate dehydrogenase and (excluding 2 patients taking aspirin) in 24-h urinary excretion of 11β-prostaglandin F2α. Overall, no change occurred in percent involvement of bone marrow by mastocytosis. Symptom scores were mild and did not change during the treatment. The 36-Item Short Form Health Survey scores for quality of life physical and mental components showed no marked changes. Tamoxifen, an older, nonhematotoxic medication, has limited activity in systemic mastocytosis at the dosage used in this study.


Primary cytomegalovirus infection, atypical Kawasaki disease, and coronary aneurysms in 2 infants.
We describe 2 infants who developed atypical Kawasaki disease and coronary aneurysms during primary cytomegalovirus infection. These observations suggest that children with coronary aneurysms and Kawasaki-like disease should be tested for cytomegalovirus. Conversely, children with unusually severe primary cytomegalovirus infection should be tested for coronary aneurysms.


Concentrations and chemical forms of heavy metals in the bulk atmospheric deposition of Beijing, China.
This study aimed to investigate concentrations and mobility of selected metals (As, Cd, Cr, Co, Cu, Mn, Ni, Pb and Zn) contained in the bulk atmospheric deposition of Beijing, China, between March 2007 and February 2008, and to evaluate the metal deposition fluxes. Beijing endured heavy atmospheric deposited metal pollution with a deposition flux of 149.28 kg km-2 year-1 from nine metals studied, and a significant increasing trend was noted from metals studied during the past 23 years. High enrichment factors (EFs) of As, Zn, Pb, Cd, and Cu manifested their major anthropogenic sources. Among which, Cd and Pb require a consistent monitoring and to be strictly controlled regarding their high mobility and ecotoxicity. Furthermore, a declined metal concentration was observed in the bulk atmospheric deposition during spring, and long-transported crustal particles sourced from East Asian dust emission belt can reasonably account for this phenomenon.


Mobilization of Environmental Toxicants Following Bariatric Surgery.
Persistent organic pollutants (POPs) are lipophilic environmental toxicants that accumulate in adipose tissue. Weight loss leads to mobilization and increased redistribution of these toxicants. Many are obesogens and endocrine disruptors. Increased exposure could pose long-term health risks. The study objective was to measure the changes in serum concentrations of lipophilic POPs during significant weight loss. This study enrolled 27 patients at a university hospital in a longitudinal, 6-month, observational study examining changes in POP blood levels after bariatric surgery. The primary outcome was the changes in the concentrations of 24 polychlorinated biphenyls (PCBs), 9 organochlorine pesticides (OCPs), 11 polybrominated diphenyl ethers, 2,2',4,4',5,5'-hexabromobiphenyl, and 4 perfluorochemicals (PFCs). Older adults (those born before 1976) had baseline levels of PCBs, OCPs, and PFCs that were two- to fivefold higher than younger adults (those born after 1976). Older adults had greater increases in PCBs, OCPs, and polybrominated diphenyl ethers associated with weight loss. Conversely, younger adults had greater increases in PFCs associated with weight loss. On average, blood POP levels increased as weight loss occurred. Although weight loss is considered beneficial, the release and redistribution of POPs to other lipid-rich organs such as the brain, kidneys, and liver warrant further investigation. Interventions should be considered to limit organ exposure to POPs when weight loss interventions are planned.


The rise of thawing drip: Freezing rate effects on ice crystallization and myowater dynamics changes.
To better reveal the formation of thawing drip, this study investigated the ice crystallization and myowater dynamics changes in frozen bovine Longissimus dorsi muscle. In ultra-fast freezing a narrow distribution of ice crystals size was observed together with higher solubility, lower surface hydrophobicity and stable second structure of myofibrillar protein. Accordingly, ultra-fast freezing samples exhibited significantly lower thaw loss (4.35 %) than slow freezing (8.22 %) after 48 h of freezing. Upon thawing, 2D T1-T2 relaxation spectra indicated a myowater redistribution, in which slow freezing led to major migration of water from immobile water to free water. Besides, T1 and T2 relaxation times showed an increasing trend with freezing process. The proton density images displayed major free water seep from myofibrils to the surface of muscle. Consequently, the water from the "reservoir" (free water) flowed into the "channel" (the widened spaces between muscle fibres), and formed into the thawing drip.


p210bcr-abl induces amoeboid motility by recruiting ADF/destrin through RhoA/ROCK1.
We previously demonstrated that the Bcr-Abl oncogene, p210(bcr-abl), through its unique GEF domain, specifically activates RhoA and induces spontaneous amoeboid motility. We intend to study the pathways downstream RhoA controlling amoeboid motility. Mouse prolymphoblastic cells (Ba/F3 cell line) expressing different forms of Bcr-Abl were embedded in 3-dimensional (3D) Matrigel to study motility and explore the effects of inhibiting Rho pathway (inhibitors and siRNAs). The phosphorylation levels of cofilin-1 and destrin were analyzed by 2-dimensional electrophoresis. Composition of Bcr-Abl signalplex in different conditions was determined by coimmunoprecipitation. Ba/F3p190 and Ba/F3 expressing a mutant form of p210(bcr-abl) (unable to activate RhoA) cells presented a spontaneous motility, but not an amoeboid type. p210(bcr-abl)-induced amoeboid motility in a 3D matrix requires isoform-specific RhoA/ROCK-1/destrin signaling. Next to the conventional Rho/ROCK/MLC/myosin pathway, this pathway is a crucial determinant for amoeboid motility, specific for the destrin isoform (and not its coexpressed homologue cofilin-1). Also, the presence of destrin (and not cofilin-1) in the p210(bcr-abl) complex is dependent on ROCK1, and this signalplex is required for amoeboid motility. This underscores isoform-specific function within the ADF/cofilin family and provides new insight into Bcr-Abl signaling to amoeboid motility and possible impact on understanding chronic myeloid leukemia progression.


The immunosuppressive tumor microenvironment in hepatocellular carcinoma.
Increasing evidence indicates the immunosuppressive nature of the local environment in tumor. The present study was focused on analyzing the immune status within hepatocellular carcinoma. In contrast to the increasing number of CD4(+) T cells, CD8(+), CD3(-)CD56(+), CD3(+)CD56(+), and gammadeltaT cells were all found to be under-represented in tumor infiltrating lymphocytes. Notably, the relative abundance of CD3(+)CD56(+) cells appeared to be correlated with patient survival. Functional analysis demonstrated that CD4(+) cells in the tumor tended to produce more IL-10 but less IFN-gamma, whereas CD8(+) cells showed impaired capacity for the production of both IFN-gamma and perforin. Consistent with previous reports, we observed a significant increase of Foxp3(+) cells in the tumor tissue. Intriguingly, although over 90% of CD4(+)CD25(high) cells were found to be Foxp3(+), the majority of Foxp3(+) cells were identified in the CD4(+)CD25(medium) and CD4(+)CD25(-) subsets. In support of its role as a negative regulator, CD4(+)CD25(high) cells suppressed the proliferation of CD4(+)CD25(-) cells isolated from the same tissues in an APC dependent manner. In conclusion, the tumor microenvironment of hepatocellular carcinoma is featured by the presence of multiple immunosuppressive factors.


Novel use of chloro-aluminum phthalocyanine assisted photodynamic therapy helps in periimplant healing among smoking patients.
To evaluate the clinical and biochemical peri‑implant parameters using chloro‑aluminum phthalocyanine-(CAP) mediated antimicrobial photodynamic therapy (PDT) as an adjunct to non-surgical mechanical debridement (NSD) in cigarette smokers (CS) and never-smokers (NS) with peri‑implantitis. Thirty-two patients with peri‑implantitis [Group CS - 16 and Group NS - 16] were recruited for the trial. The study participants underwent two therapies: PDT + NSD and NSD alone. The clinical peri‑implant parameters assessed in this clinical trial were bleeding on probing (BP), crestal bone loss (CBL) peri‑implant pocket depth (PD), and peri‑implant plaque scores (PS), respectively. Peri-implant crevicular fluid was sampled and the quantification of interleukin (IL)-1β and tumor necrosis factor-alpha (TNF-α) was performed using enzyme linked immunosorbent assay (ELISA). BP was significantly reduced at 6 months after PDT+NSD in CS groups. Mean PD significantly reduced after both PDT+NSD and NSD subgroups within both NS and CS groups. Only at 6 months did PDT+NSD showed statistically significantly reduced IL-1β levels in the NS group. TNF-α levels significantly reduced in CS group with PDT+NSD and NSD alone at both 3 months and 6 months follow up. CAP-assisted PDT helped to improve the clinical and cytokine levels after non-surgical peri‑implant mechanical debridement in treating peri‑implantitis patients in smokers.


Ontogeny and expression of non-MHC-restricted T cell antigen-binding molecules by thymocytes and peripheral T cell subsets.
The ontogeny of T cells which express major histocompatibility complex (MHC)-unrestricted T cell antigen-binding molecules (TABM) on the cell membrane was investigated. We used a rabbit anti-mouse TABM antiserum to investigate the expression of TABM by subsets of adult thymocytes, peripheral T cells, and thymocytes during gestation. TABM are expressed by CD4+, CD8-, CD4+, CD8+ thymocytes and single-positive thymocytes. During gestation, TABM are expressed as early as Day 16, and at birth the expression of TABM on thymocytes has reached adult levels. Data are also presented which suggest that the expression of membrane TABM (mTABM) on peripheral T cells can be upregulated during T cell activation. The results suggest that TABM are expressed by different T cell subsets and that TABM+ cells may utilize the same intrathymic developmental pathway as that of T cells which express the alpha/beta T cell receptor.


Use of denaturing HPLC to provide efficient detection of mutations causing guanidinoacetate methyltransferase deficiency.
Guanidinoacetate methyltransferase (GAMT) deficiency is an autosomal recessive error of creatine synthesis characterized by cerebral creatine deficiency, accumulation of guanidinoacetate, mental retardation, epilepsy, and extrapyramidal symptoms. To date, 14 mutations of the GAMT gene in 27 patients have been reported. Mutation analysis was done using direct sequencing of PCR products and denaturing gradient gel electrophoresis in combination with direct sequencing. In contrast, we evaluated the efficiency of a newly developed DHPLC method to detect mutations in the GAMT gene by analysing DNA from 14 GAMT patients with known mutations. PCR amplification of both patient and control DNA was followed by formation of homoduplices and heteroduplices, and their detection by DHPLC. DHPLC identified all mutations tested and is the preferred choice of analytical method.


Effectiveness of eight or more antenatal contacts on health facility delivery and early postnatal care in low- and middle-income countries: a propensity score matching.
Despite progress in reducing maternal and child mortality, many low- and middle-income countries (LMICs) still experience an unacceptably high level of the problem. The World Health Organization (WHO) recently recommended pregnant women should have at least eight antenatal care visits (ANC8+) with a trained healthcare provider as a key strategy to promote pregnant women's health. Antenatal care is an imperative factor for subsequent maternal healthcare utilization such as health facility delivery and early postnatal care (EPNC). This study aimed to examine the net impact of ANC8+ visits on health facility delivery and EPNC in LMICs using a propensity score matching analysis. We used the recent Demographic and Health Survey (DHS) datasets from 19 LMICs. Women of reproductive age (15-49 years) who had given birth within 1 year preceding the survey were included. A propensity score matching analysis was employed to assess the net impact of eight or more antenatal care visits on health facility delivery and early postnatal care. After matching the covariates, women who attended ANC8+ visits had a 14% (ATT = 0.14) higher chance of having their delivery at health facilities compared with women who attended less than eight ANC visits. This study further revealed that women who had ANC8+ visits were associated with a 10% (ATT = 0.10) higher probability of early PNC compared with their counterparts. This study confirmed that ANC8+ visits significantly increased the likelihood of health facility-based delivery and early PNC utilization in LMICs. These findings call for public health programs to focus on pregnant women attending adequate ANC visits (according to revised WHO recommendation) as our study indicates that ANC8+ visits significantly improved the chances of subsequent care.


Influence of intensity of illumination during the light period on diurnal variations of pineal indoles in rats and mice.
The diurnal variations of pineal indoles in New-Zealand Black (NZB) mice and Wistar rats were determined by HPLC fluorometry. Pineal N-acetylserotonin (NAc5HT) in NZB mice showed a marked diurnal variation, but no melatonin (MLT) was detected at any time of day or night. The night-time increases of NAc5HT in NZB mice and MLT in Wistar rats were delayed about one hour in animals kept under illumination of high intensity (2000 lux) during the light period (12 h) compared with those in animals kept under illumination of low intensity (10 lux). These results suggest that the intensity of illumination during the light period affects the diurnal rhythms of pineal indoles in experimental animals.


Remediation of PFAS-Contaminated Soil and Granular Activated Carbon by Smoldering Combustion.
This study explored smoldering combustion for remediating polyfluoroalkyl substance (PFAS)-impacted granular activated carbon (GAC) and PFAS-contaminated soil. GAC, both fresh and PFAS-loaded, was employed as the supplemental fuel supporting smoldering in mixtures with sand (≈175 mg PFAS/kg GAC-sand), with PFAS-spiked, laboratory-constructed soil (≈4 mg PFAS/kg soil), and with a PFAS-impacted field soil (≈0.2 mg PFAS/kg soil). The fate of PFAS and fluorine was quantified with soil and emission analyses, including targeted PFAS and suspect screening as well as hydrogen fluoride and total fluorine. Results demonstrated that exceeding 35 g GAC/kg soil resulted in self-sustained smoldering with temperatures exceeding 900 °C. Post-treatment PFAS concentrations of the treated soil were near (2 experiments) or below (7 experiments) detection limits (0.0004 mg/kg). Further, 44% of the initial PFAS on GAC underwent full destruction, compared to 16% of the PFAS on soil. Less than 1% of the initial PFAS contamination on GAC or soil was emitted as PFAS in the quantifiable analytical suite. Results suggest that the rest were emitted as altered, shorter-chain PFAS and volatile fluorinated compounds, which were scrubbed effectively with GAC. Total organic fluorine analysis proved useful for PFAS-loaded GAC in sand; however, analyzing soils suffered from interference from non-PFAS. Overall, this study demonstrated that smoldering has significant potential as an effective remediation technique for PFAS-impacted soils and PFAS-laden GAC.


Traditional approaches to estimate length at first maturity (L50 ) retrieve better results than alternative ones in a Neotropical heptapterid.
Length at first maturity (L50 ) is an important tool for the management and conservation of fish populations. Traditional approaches based on macroscopic and microscopic maturity staging exhibit high accuracy and precision, while alternative approaches (e.g., Ig -based staging, stanza changing point) are less resource-demanding. Herein, we compare four approaches to estimate L50 in a population of the heptapterid Rhamdioglanis transfasciatus from Atlantic Forest streams. Fish were sampled monthly during a year by using electrofishing. We measured the length (cm), mass (g), and gonad mass (g) of each specimen, then classified their maturity status macroscopically and microscopically. Alternative approaches were strongly discordant from traditional ones. Logistic curves considering mature individuals as those displaying at least 1% of the maximum Ig in the sample greatly underestimated L50 for females and overestimated L50 for males. The stanza changing point derived from the polyphasic growth model underestimated L50 in both cases. Despite the increasing development of less onerous approaches, it seems that they are not suitable for all fish populations and the requirements to use such approaches demand further investigation.


A novel method for the preparation of biodegradable microspheres for protein drug delivery.
Microspheres are potential candidates for the protein drug delivery. In this work, we prepared polymer-coated starch/bovine serum albumin (BSA) microspheres using co-axial electrohydrodynamic atomization (CEHDA). First, starch solution in dimethyl sulphoxide (DMSO) was prepared and then an aqueous solution of BSA was added to it to make a starch-BSA solution. Subsequently, this solution was made to flow through the inner capillary, while the polymer, polydimethylsiloxane (PDMS), flowed through the outer capillary. On collection, filtration and subsequent drying, near-monodisperse microspheres of 5-6microm in size were obtained. The microspheres were characterized by Fourier-transform infrared (FT-IR) spectroscopy and scanning electron microscopy. Cumulative BSA release was investigated by UV spectroscopy. BSA structure and activity was preserved in the microspheres and its release in 0.01M phosphate buffered saline (PBS) was studied over a period of 8 days. There was an initial burst with 32wt% of total BSA released in 2h. Overall 75wt% of BSA was released over a 7 day period.


Capabilities and limitations of probe methods for the microanalysis of chemical elements in biology: a brief introduction.
This article reviews some of the technical problems involved in achieving high-resolution artefact-free quantitative microprobe analyses of biological specimens, and points to some of the techniques now available to overcome these problems.


A validated preoperative risk prediction tool for extended inpatient length of stay following anatomic or reverse total shoulder arthroplasty.
Recent work has shown inpatient length of stay (LOS) following shoulder arthroplasty to hold the second strongest association with overall cost (after implant cost itself). In particular, a preoperative understanding for the patients at risk of extended inpatient stays (≥3 days) can allow for counseling, optimization, and anticipating postoperative adverse events. A multicenter retrospective review was performed of 5410 anatomic (52%) and reverse (48%) total shoulder arthroplasties done at 2 large, tertiary referral health systems. The primary outcome was extended inpatient LOS of at least 3 days, and over 40 preoperative sociodemographic and comorbidity factors were tested for their predictive ability in a multivariable logistic regression model based on the patient cohort from institution 1 (derivation, N = 1773). External validation was performed using the patient cohort from institution 2 (validation, N = 3637), including area under the receiver operator characteristic curve (AUC), sensitivity, specificity, and positive and negative predictive values. A total of 814 patients, including 318 patients (18%) in the derivation cohort and 496 patients (14%) in the validation cohort, experienced an extended inpatient LOS of at least 3 days. Four hundred forty-five (55%) were discharged to a skilled nursing or rehabilitation facility. Following parameter selection, a multivariable logistic regression model based on the derivation cohort (institution 1) demonstrated excellent preliminary accuracy (AUC: 0.826), with minimal decrease in accuracy under external validation when tested against the patients from institution 2 (AUC: 0.816). The predictive model was composed of only preoperative factors, in descending predictive importance as follows: age, marital status, fracture case, ASA (American Society of Anesthesiologists) score, paralysis, electrolyte disorder, body mass index, gender, neurologic disease, coagulation deficiency, diabetes, chronic pulmonary disease, peripheral vascular disease, alcohol dependence, psychoses, smoking status, and revision case. A freely-available, preoperative online clinical decision tool for extended inpatient LOS (≥ 3 days) after shoulder arthroplasty reaches excellent predictive accuracy under external validation. As a result, this tool merits consideration for clinical implementation, as many risk factors are potentially modifiable as part of a preoperative optimization strategy.


WITHDRAWN: The clinical application of an innovative tracheotomy fixation belt in critically ill patients.
Ahead of Print article withdrawn by publisher.


Infection rate and genetic diversity of Giardia duodenalis in pet and stray dogs in Henan Province, China.
Giardia duodenalis is an important protozoan parasite that is known to be zoonotic. To assess the potential zoonotic transmission of giardiasis from dogs and to identify genetic diversity of G. duodenalis in dog populations, we examined the infection rate and genotypes of G. duodenalis in both pet dogs (from pet dog farms, pet shops, pet hospitals, pet markets) and stray dogs of different ages in Henan Province, China. A total of 940 fresh fecal specimens were collected from 2007 to 2013 in Henan Province. The overall infection rate of G. duodenalis was 14.3% (134/940) as determined by microscopy, with the highest infection rate (17.3%) observed in dogs from shelters. Young dogs were more likely to be infected with G. duodenalis than adult dogs, and G. duodenalis cysts were found more frequently in diarrheic dogs. All G. duodenalis-positive isolates were characterized at the triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh), and β-giardin (bg) loci, and 37, 51, and 48 sequences were obtained, respectively. The dog-specific assemblages C and D were identified using multi-locus sequence analysis. Six novel sequences of the tpi locus, one novel sequence of the gdh locus and two novel sequences of the bg locus were detected among the G. duodenalis assemblage C isolates, while two novel sequences of the gdh locus were found among the G. duodenalis assemblage D isolates. Our data indicate that G. duodenalis is a common parasite and cause of diarrheal disease in dogs in Henan Province. However, there was no evidence for zoonotic G. duodenalis assemblages in the study population.


Anticardiolipin antibodies in proliferative diabetic retinopathy: An additional risk factor.
To report the prevalence of anticardiolipin antibodies in patients with proliferative diabetic retinopathy (PDR) having high-risk criteria (HRC). Diabetic patients having PDR with HRC and diabetics free of retinopathy were compared for the presence of anticardiolipin antibodies. Among the 34 patients, 6 (17.7%) of diabetics having PDR with HRC were positive for anticardiolipin antibodies. There was no significant association of aCL antibodies with sex or type of diabetes. Using Pearson's correlation test, no significant associations of aCL antibodies with duration of diabetes or age of patients were found. All patients who were positive for anticardiolipin antibodies had PDR with HRC. The difference was statistically significant. Presence of anticardiolipin antibodies may represent an additional risk factor for PDR.


Distribution of polybrominated diphenyl ethers (PBDEs) in feather and muscle of the birds of prey from Beijing, China.
Polybrominated diphenyl ethers (PBDEs) in the feather and muscle of common kestrels (Falco tinnunculus), eagle owls (Bubo bubo) and little owls (Athene noctua) collected from Beijing, China were investigated. The concentrations of ∑23PBDEs in the muscle and feather of all the birds of prey ranged from 46 to 7.77 × 104 ng/g lipid weight (lw; median 241 ng/g lw) and 1.50-191 ng/g dry weight (dw; median 21.1 ng/g dw), respectively. Higher brominated congeners, e.g., BDE-209, -153, -207 and -196 were the dominant congeners in both feather and muscle. However, the concentrations of lower brominated congeners in feather were significantly correlated with those in muscle (p < 0.05), which suggested that feather could efficiently reflect lower brominated BDEs in the internal tissue of birds of prey. The calculated hazard quotients (HQs) > 10 in common kestrel and little owl suggested that the high levels of PBDEs posed potentially high risk to some birds of prey living in Beijing area.


Use and Safety of Anthroposophic Medicinal Products: An Analysis of 44,662 Patients from the EvaMed Pharmacovigilance Network.
There is a need for data on the clinical safety of anthroposophic medicinal products (AMPs). The main objective of this analysis was to determine the frequency of adverse drug reactions (ADRs) to AMPs, relative to the number of AMP prescriptions. EvaMed (Evaluation of Anthroposophic Medicine) was a prospective pharmacovigilance study with the patients of 38 physicians in outpatient care in Germany. Diagnoses and prescriptions were extracted from the electronic medical records. All physicians documented ADRs of Grades III-IV and serious ADRs, seven 'prescriber physicians' also documented non-serious ADRs of any intensity. Patients were eligible for this analysis if they had one or more AMP prescription in the years 2001-2010, followed by one or more physician visit. A total of 44,662 patients with 311,731 AMP prescriptions, comprising 1722 different AMPs, were included. One hundred ADRs to AMPs occurred, caused by 83 different AMPs. ADR intensity was mild, moderate, and severe in 50% (n = 50/100), 43%, and 7% of cases, respectively; one ADR was serious. Among patients of prescriber physicians, ADRs of any intensity occurred in 0.071% (n = 67/94,734) of AMP prescriptions and in 0.502% (n = 65/12,956) of patients prescribed AMPs. In subgroup analyses according to age, specific AMPs or AMP groups, dosage forms, and concentrations (altogether 11 groups), the highest ADR frequency was 0.290% of prescriptions (for one specific AMP). Among all patients, serious ADRs occurred in 0.0003% (n = 1/311,731) of prescriptions and 0.0022% (n = 1/44,662) of patients. In this analysis from a large sample, ADRs to AMP therapy in outpatient care were rare; ADRs of high intensity as well as serious ADRs were very rare.


Wall stress determines systolic and diastolic function--Characteristics of heart failure.
Heart failure can be caused by systolic or diastolic dysfunction. Diagnosing diastolic dysfunction remains challenging, although several criteria have been identified. Ventricular wall stress is crucially involved. It is hypothesized whether increased end-diastolic and end-systolic ventricular wall stress as assessed by the wall stress index is associated with cardiac dysfunction and thus provide novel diagnostic criteria. 1050 consecutive patients with suspected non-ischemic heart failure covering a broad spectrum from normal to severely impaired cardiac function were observed. Cardiac magnetic resonance imaging was performed to assess left ventricular (LV) volumes, myocardial mass, peak ejection (PER) and filling rate (PFR). A reduced PFR was found in 348 patients (33.1%), which resulted from 275 of 422 patients (65.2%) with reduced and from 73 of 628 patients (11.6%) with preserved LVEF (p<0.0001). Increased LV volume and mass was correlated with reduced PER and PFR (p<0.0001). Increased end-diastolic wall stress was the strongest predictor of a reduced PER (OR 4.5 [2.6 to 7.8], p<0.0001) and increased end-systolic wall stress predicted a reduced PFR (OR 1.2 [1.1 to 1.3], p<0.0001). Increased end-systolic wall stress was correlated with increased pulmonary pressure (p<0.0001). Normal end-systolic wall stress<18 kPa had a favorable predictive value for the absence of an impaired filling and increased pulmonary capillary pressure. Increased end-diastolic wall stress precedes a reduced ventricular ejection rate and increased end-systolic wall stress determines an impaired diastolic filling. It is thus suggested to add assessment of ventricular wall stress as diagnostic criterion of heart failure.


Mechanical stretch induced interleukin-18 (IL-18) expression through Angiotensin subtype 1 receptor (AT1R) and endothelin-1 in cardiomyocytes.
Interleukin-18 (IL-18) is a proinflammatory cytokine with multiple biological functions. We and others have demonstrated that an increased level of circulating IL-18 is one of the risk factors for cardiovascular diseases. Endothelin-1 (ET-1) has been reported to be a potent hypertrophy-promoting factor through RhoA and Rho-Kinase. Mechanical stretch induces a hypertrophic response, partly through the production of ET-1 through Endothelin A receptor (ETAR). Moreover, it has also been reported that mechanical stretch induces cardiac hypertrophy through Angiotensin subtype 1 receptor (AT1R). However, the mechanism by which the IL-18 gene expression is regulated in cardiomyocytes has not yet been fully understood. This study was designed to elucidate the functional significance of IL-18 gene expression in response to mechanical stretch. Neonatal rat cardiomyocytes cultured on silicone dishes were subjected to stretch. The moderate 20% mechanical stretch resulted in the elevation of IL-18 expression in a time-dependent manner with the maximal level achieved 36 hours after the stretch. Olmesartan, AT1R antagonist inhibited stretch-induced IL-18 expression. ETAR blockade BQ123 inhibited stretch-induced IL-18 expression. However, the Endothelin B receptor (ETBR) receptor blockade BQ788 did not inhibit this reaction. ET-1 induced IL-18 expression, with a peak induction after 4 hours of incubation. These results might suggest that stretch stimulation of cardiomyocytes induced ET-1 and, subsequently, ET-1 up-regulated the IL-18 expression. Furthermore, Fasudil, a Rho-Kinase inhibitor, and Simvastatin, a HMG-CoA reductase inhibitor, led to a significant reduction in mechanical stretch-induced IL-18 expression. These results indicated, for the first time, that IL-18 expression is induced by mechanical stretch in cardiomyocytes via the ETAR, AT1R, and the Rho/Rho-K pathways. The induction of IL-18 from cardiomyocytes by mechanical stress might cause the deterioration of cardiac functions in autocrine and paracrine fashion. The inhibition of IL-18 expression induced by mechanical stress might be one of the mechanisms that account for the beneficial cardiovascular effects of AT1R antagonist, ETAR blockade, Statin, and Rho-Kinase inhibitor.


Acute stroke treatment and outcome in the oldest old (90 years and older) at a tertiary care medical centre in Germany-a retrospective study showing safety and efficacy in this particular patient population.
Stroke is among the most common causes of death and disability worldwide. Despite the relevance of stroke-related disease burden, which is constantly increasing due to the demographic change in industrialized countries with an ageing population and consecutively an increase in age-associated diseases, there is sparse evidence concerning acute stroke treatment and treatment-related outcome in the elderly patient group. This retrospective study aimed at analysing patient characteristics, therapy-related complications and functional outcome in stroke patients aged 90 years or older who underwent acute stroke treatment (i.e. intravenous thrombolysis, mechanical thrombectomy, or both). We identified files of all inpatient stays at the Department of Neurology at Saarland University Medical Center (tertiary care level with a comprehensive stroke unit) between June 2011 and December 2018 and filtered for subjects aged 90 years or older at the time of admission. We reviewed patient files for demographic data, symptoms upon admission, (main) diagnoses, comorbidities, and administered therapies. For patients admitted due to acute stroke we reviewed files for therapy-related complications and functional outcome. We compared the modified Rankin scale (mRS) scores upon admission and at discharge for these patients. We identified 566 inpatient stays of subjects aged 90 years or older. Three hundred sixty-seven of the 566 patients (64.8%) were admitted and discharged due to symptoms indicative of stroke. Two hundred eleven patients received a diagnosis of ischaemic stroke. These 211 patients were analysed subsequently. Sixty-four patients qualified for acute stroke treatment (intravenous thrombolysis n = 22, mechanical thrombectomy n = 26, intravenous thrombolysis followed by mechanical thrombectomy n = 16) and showed a significant improvement in their functional status as measured by change in mRS score (admission vs. discharge, p 0.001) with 7 (10.9%) observed potentially therapy-related complications (relevant drop in haemoglobin n = 2, subarachnoidal haemorrhage n = 1, cerebral haemorrhage n = 3, extracranial bleeding n = 1). One intravenous thrombolysis was stopped because of an uncontrollable hypertensive crisis. Patients who did not qualify for these treatments (including those declining acute treatment) did not show a change of their functional status between admission and discharge (p 0.064). Our data indicate that acute stroke treatment is effective and safe in the oldest old. Age alone is no criterion to withhold an acute intervention even in oldest old stroke patients.


Exercise responses in patients with chronically high creatine kinase levels.
Elevated serum creatine kinase (CK) is often taken to reflect muscle disease, but many individuals have elevated CK without a specific diagnosis. How elevated CK reflects muscle metabolism during exercise is not known. Participants (46 men, 48 women) underwent incremental exercise testing to assess aerobic performance, cardiovascular response, and ventilatory response. Serum lactate, ammonia, and CK were measured at rest, 4 minutes into exercise, and 2 minutes into recovery. High-CK and control subjects demonstrated similar aerobic capacities and cardiovascular responses to incremental exercise. Those with CK ≥ 300 U/L exhibited significantly higher lactate and ammonia levels after maximal exercise, together with increased ventilatory responses, whereas those with CK ≥200 U/L but ≤ 300 U/L did not. We recommend measurement of lactate and ammonia profiles during a maximal incremental exercise protocol to help identify patients who warrant muscle biopsy to rule out myopathy. Muscle Nerve 56: 264-270, 2017.


Envelope protein changes, autoagglutination, sensitivity to hydrophobic agents and a conditional division lesion in Escherichia coli strains carrying ColV virulence plasmids.
The presence of the virulence plasmids ColV,I-K94 or ColV-K30 in Escherichia coli produces a number of cell membrane and envelope changes. The most striking of these are (1) the presence of the 33K VmpA outer membrane protein and (2) the ColV-associated occurrence of autoagglutination. The VmpA protein is a plasmid-encoded outer membrane protein which is synthesized from a larger precursor. It is distinct from the chromosomally-encoded OmpA protein but resembles it in a few respects. The VmpA protein does not appear to be involved in colicin synthesis or immunity, or in plasmid transfer. This protein was found in 6 out of 8 new ColV+ isolates, but not in 2 ColIa+ strains. ColV-induced autoagglutination occurred for strains grown in static culture at 37 but not at 25 degrees C. Detergents prevented agglutination, as did the presence in a ColV+ strain of a fi+ plasmid, ColB. Autoagglutination may be a virulence phenotype. Associated with the ability of ColV+ bacteria to agglutinate was inhibition of motility. ColV+ bacteria also showed changes in envelope permeability indicated by inhibitor sensitivity and by a ColV-associated suppression of the lac Y lesion. Some ColV,I-K94+ strains showed a mucoid colonial phenotype and this ability to form mucoid colonies was efficiently transferred with ColV but apparently not without it. The mucoid ColV+ strains resembled lon mutants in UV-sensitivity, division behaviour and sensitivity to lambda phage.


Diagnostic and therapeutic application of thoracic epidural in a child with intractable cardiac arrhythmias: A case report.
A 3-year-old boy presented with episodes of uneasiness and transient loss of consciousness. Atrial tachyarrhythmias with rapid ventricular rate was diagnosed and initially unsuccessfully treated with oral antiarrhythmic drugs. Subsequent Holter monitoring revealed ventricular arrhythmias. Despite pharmacologic treatment, he needed numerous cardioversions. Surgical sympathectomy was planned. Initially, sympathectomy was achieved using a continuous high thoracic epidural block and was performed to ascertain the efficacy of the thoracic sympathectomy. This successfully reduced the ventricular arrhythmias and the need for antiarrhythmic agents. The epidural infusion was also used for pain relief following the subsequent surgical sympathectomy.


Qualitative versus quantitative methods in psychiatric research.
Qualitative studies are gaining their credibility after a period of being misinterpreted as "not being quantitative." Qualitative method is a broad umbrella term for research methodologies that describe and explain individuals' experiences, behaviors, interactions, and social contexts. In-depth interview, focus groups, and participant observation are among the qualitative methods of inquiry commonly used in psychiatry. Researchers measure the frequency of occurring events using quantitative methods; however, qualitative methods provide a broader understanding and a more thorough reasoning behind the event. Hence, it is considered to be of special importance in psychiatry. Besides hypothesis generation in earlier phases of the research, qualitative methods can be employed in questionnaire design, diagnostic criteria establishment, feasibility studies, as well as studies of attitude and beliefs. Animal models are another area that qualitative methods can be employed, especially when naturalistic observation of animal behavior is important. However, since qualitative results can be researcher's own view, they need to be statistically confirmed, quantitative methods. The tendency to combine both qualitative and quantitative methods as complementary methods has emerged over recent years. By applying both methods of research, scientists can take advantage of interpretative characteristics of qualitative methods as well as experimental dimensions of quantitative methods.


Coordinated morphogenesis of neurons and glia.
Glia adopt remarkable shapes that are tightly coordinated with the morphologies of their neuronal partners. To achieve these precise shapes, glia and neurons exhibit coordinated morphological changes on the time scale of minutes and on size scales ranging from nanometers to hundreds of microns. Here, we review recent studies that reveal the highly dynamic, localized morphological changes of mammalian neuron-glia contacts. We then explore the power of Drosophila and C. elegans models to study coordinated changes at defined neuron-glia contacts, highlighting the use of innovative genetic and imaging tools to uncover the molecular mechanisms responsible for coordinated morphogenesis of neurons and glia.


Association of Prophylaxis and Length of Stay With Venous Thromboembolism in Abdominopelvic Surgery.
Extended venous thromboembolism prophylaxis (eVTEp) is recommended for select patients who have undergone major abdominopelvic surgery to prevent postdischarge venous thromboembolism (pdVTE). Criteria for selection of these patients are untested for this purpose and may be ineffective. To address this gap, we investigated the effectiveness of eVTEp on pdVTE rates. A retrospective cohort study of patients undergoing abdominopelvic surgery from January 2016 to February 2020 was performed using data from the Michigan Surgical Quality Collaborative. pdVTE was the main outcome. Our exposure variable, eVTEp, was compared dichotomously. Length of stay (LOS) was compared categorically using clinically relevant groups. Age, race, cancer occurrence, inflammatory bowel disease, surgical approach, and surgical time were covariates among other variables. Descriptive statistics, propensity score matching, and multivariable logistic regression were performed to compare pdVTE rates. A total of 45,637 patients underwent abdominopelvic surgery. Of which, 3063 (6.71%) were prescribed eVTEp. Two hundred eighty-five (0.62%) had pdVTE. Of the 285, 59 (21%) patients received eVTEp, while 226 (79%) patients did not. After propensity score matching, multivariable logistic regression analysis showed pdVTE was associated with eVTEp and LOS of 5 d or more (P < 0.001). eVTEp was not associated with LOS. Further analysis showed increased risk of pdVTE with increasing LOS independent of prescription of eVTEp based on known risk factors. pdVTE was associated with increasing LOS but not with other VTE risk factors after propensity score matching. Current guidelines for eVTEp do not include LOS. Our findings suggest that LOS >5 d should be added to the criteria for eVTEp.


Transformation of Aspergillus niger with the homologous nitrate reductase gene.
A homologous transformation for Aspergillus niger was developed based on the nitrate reductase structural gene niaD. This system offered certain advantages over existing A. niger systems, such as the ease of recipient mutant isolation, absence of abortive transformants, convenient enzyme assay, ease of transformant stability testing, and complete absence of background growth. Transformation frequencies of up to 100 transformants per microgram DNA were obtained with the vector pSTA10 which carries the niaD gene of A. niger. Southern blotting analysis indicated that vector DNA had integrated into the genome of A. niger. Mitotic stability studies demonstrated that while some transformants were as stable as the wild-type (wt), others were markedly less so. No correlation was seen between plasmid integration, mitotic stability and nitrate reductase activity, which was markedly different from wt in only three of the transformants examined.


Inherited disorders of vitamin B12 metabolism.
Inherited disorders of vitamin B12 include those which involve the inability of the vitamin to be absorbed from the gut and transported to the appropriate tissues, and those in which the vitamin is not utilised by target cells. The former include intrinsic factor abnormalities, selective malabsorption of vitamin B12 with proteinuria, and deficiencies of transcobalamin I and transcobalamin II. The latter include a defect in the release of free vitamin B12 from lysosomes (cblF), and defects in the formation of both vitamin B12 cofactors (cblC, cblD) or of adenosyl-B12 (cblA, cblB) or methyl-B12 alone (cblE, CblE variant). This article reviews the major clinical manifestations of these diseases, and provides an approach to the diagnosis of transcobalamin II deficiency and the cbl mutations using cultured cells.


Sexually dimorphic distribution of sst2A somatostatin receptors on growth hormone-releasing hormone neurons in mice.
The pulsatile pattern of GH secretion exhibits sexual dimorphism that is likely to depend on somatostatin (SRIH) effects on somatoliberin (GHRH) neurons in the hypothalamus. Using transgenic GHRH-enhanced green fluorescent protein (eGFP) mice, no difference in the total number of GHRH-eGFP neurons or change in somatostatin receptor sst2 and SRIH mRNA levels in ventromedial hypothalamic nucleus-arcuate nucleus and periventricular nucleus regions and GHRH mRNA levels in the ventromedial hypothalamic-arcuate region were observed between male and female mice. However, the percentage of GHRH-eGFP neurons bearing sst2A receptors reached 78% in female vs. 26% in male GHRH-eGFP mice (P < 0.02). This sex difference in sst2A distribution on GHRH neurons may play an important role in the sexually differentiated pattern of GH secretion.


Rodents as seed dispersers in a heath - oak wood succession.
In a Danish heathland invasion of oak shrub has taken place, the succession rate being approximately 300 m during the last 100 years. The colonisation has occurred in steps related to the delay time between seedling stage and fertility stage. Seedlings are often found in clusters originating from caches probably made by seed-eating rodents. Apodemus sylvaticus, A. flavicollis and Clethrionomys glareoles. These rodents reached autumn densities of 25-50 individuals per ha. Radioactive acorns were scatterhoarded by the rodents, which mainly deposited the acorns singly up to at least 34 m from the oak shrub (mean 15.3±8.2 m), and preferably under Empetrum nigrum mats in the walls of runways Seedlings originating from radioactive acorns were found next summer at distances of 4-37 m from the oak shrub. In early summer caches containing new seedlings had a mean size of 2.0±2.2 acorns, range 1-16; mean distance of seedlings to nearest crown projection was 24.0±23.6 m, range 1-137. Thus, rodent acorn dispersal can explain the observed succession rate of oaks into the heathland.


Biomonitoring environmental contaminants near a municipal solid-waste combustor: a decade later.
During the 1999 nesting season, eggs and nestlings of anhingas (Anhinga anhinga) and white ibises (Eudocimus albus) were collected from a colony located on the site of a municipal solid-waste (MSW) combustor and analyzed for residues of chemicals potentially released from this facility. Concentrations of most residues, including tetrachlorodibenzo-p-dioxin (TCDD). tetrachlorodibenzofuran (TCDF), arsenic, beryllium, cadmium and lead, were at levels comparable to those found during a similar survey done in 1989, prior to facility start-up. Nickel residues were detected only sporadically in anhingas and in eggs from ibises, and decreased significantly in concentration in ibis nestlings in 1999 compared to 1989. While concentrations of mercury in anhinga nestlings and in eggs of both species were comparable to 1989 levels, its concentration was significantly greater in ibis nestlings in 1999. However, levels of mercury in the ibises remained relatively low when compared to birds from other areas of Florida and did not appear to represent a health threat. While lead did not increase in 1999, its concentration in ibis nestlings remained a concern. The most notable temporal trend observed in birds at this site was a general monotonic decrease in levels of selenium residues during the 10-year monitoring period.


First nosocomial outbreak of Pseudomonas aeruginosa producing an integron-borne metallo-beta-lactamase (VIM-2) in the United States.
Carbapenemases are rare in the United States. This is the first report of a United States nosocomial outbreak of pan-resistant Pseudomonas aeruginosa infections due to an integron-borne metallo-beta-lactamase, VIM-2. This emergence of carbapenemases on mobile genetic elements in the United States warrants surveillance.


Transneuronal mapping of the CNS network controlling sympathetic outflow to the rat thymus.
The thymus is a primary immune organ that is essential for the development of functional T cells. The thymus receives sympathetic innervation, and thymocytes and thymic epithelial cells express functional adrenergic receptors. In this study, we employed retrograde, transneuronal virus tracing to identify the CNS cell groups that regulate sympathetic outflow to the thymus. Pseudorabies virus (PRV) was injected into the thymus, and the pattern of PRV infection in sympathetic regulatory centers of the CNS was determined at 72 and 120 h post-inoculation. PRV infection within the CNS first appeared within the spinal cord at 72 h post-inoculation and was confined to neurons within the intermediolateral cell column at levels T1-T7. At 120 h post-inoculation infection had spread within the spinal cord to include the central autonomic nucleus, intercalated cell nucleus and light infection within the cells of the lateral funiculus. Within the brain, PRV positive cells were found within nuclei of the medulla oblongata, pons and hypothalamus. Infection in the hypothalamus was observed within the arcuate nucleus, dorsal, lateral, and posterior hypothalamus and in all parvicellular subdivisions of the paraventricular hypothalamic nucleus. None of the infected animals exhibited labeling of the dorsal motor nucleus of the vagus. In summary, this study provides the first anatomic map of CNS neurons involved in control of sympathetic outflow to the thymus.


Beneficial effects of whole-body vibration exercise for brain disorders in experimental studies with animal models: a systematic review.
Brain disorders have been a health challenge and is increasing over the years. Early diagnosis and interventions are considered essential strategies to treat patients at risk of brain disease. Physical exercise has shown to be beneficial for patients with brain diseases. A type of exercise intervention known as whole-body vibration (WBV) exercise gained increasing interest. During WBV, mechanical vibrations, produced by a vibrating platform are transmitted, to the body. The purpose of the current review was to summarize the effects of WBV exercise on brain function and behavior in experimental studies with animal models. Searches were performed in EMBASE, PubMed, Scopus and Web of Science including publications from 1960 to July 2021, using the keywords "whole body vibration" AND (animal or mice or mouse or rat or rodent). From 1284 hits, 20 papers were selected. Rats were the main animal model used (75%) followed by mice (20%) and porcine model (5%), 16 studies used males species and 4 females. The risk of bias, accessed with the SYRCLE Risk of Bias tool, indicated that none of the studies fulfilled all methodological criteria, resulting in possible bias. Despite heterogeneity, the results suggest beneficial effects of WBV exercise on brain functioning, mainly related to motor performance, coordination, behavioral control, neuronal plasticity and synapse function. In conclusion, the findings observed in animal studies justifies continued clinical research regarding the effectiveness and potential of WBV for the treatment of various types of brain disorders such as trauma, developmental disorders, neurogenetic diseases and other neurological diseases.


Villous oedema of the placenta: a clinicopathological study.
Villous oedema was observed in 259 placentae among 1925 consecutive singleton pregnancies of greater than 19 weeks gestation. It was present in 11 per cent of term placentae in which significant associations with fetal and neonatal death (P less than 0.03), and absence of maternal cigarette smoking (P less than 0.002) were found. In preterm placentae, the oedema was usually more severe, and its prevalence increased from 20 per cent for 33-37 weeks to 40 per cent for less than 33 weeks. Our analysis showed that for a given gestational age, villous oedema was not significantly related to chorioamnionistis, Apgar scores of less than 7 at 1 and 5 min, or neonatal death, an exception was for 33-37 weeks gestation, in the absence of chorioamnionitis, villous oedema was associated with low 1 min Apgar score. Immature intermediate villi are present in premature placentae as a normal developmental stage and in dysmature placentae as a result of villous maldevelopment. Since villous oedema closely resembles the 'stromal channels' in this villous type and shows significant association with prematurity and villous dysmaturity, we postulate that villous oedema is a lesion primarily of the immature intermediate villi. Both fetal and maternal factors are involved in its pathogenesis.


Initial experience with factor-Xa inhibition in percutaneous coronary intervention: the XaNADU-PCI Pilot.
Direct factor (F)Xa inhibition is an attractive method to limit thrombotic complications during percutaneous coronary intervention (PCI). To investigate drug levels achieved, effect on coagulation markers, and preliminary efficacy and safety of several doses of DX-9065a, an intravenous, small molecule, direct, reversible FXa inhibitor during PCI. Patients undergoing elective, native-vessel PCI (n = 175) were randomized 4 : 1 to open-label DX-9065a or heparin in one of four sequential stages. DX-9065a regimens in stages I-III were designed to achieve concentrations of > 100 ng mL-1, > 75 ng mL-1, and > 150 ng mL-1. Stage IV used the stage III regimen but included patients recently given heparin. At 15 min median (minimum) DX-9065a plasma levels were 192 (176), 122 (117), 334 (221), and 429 (231) ng mL-1 in stages I-IV, respectively. Median whole-blood international normalized ratios (INRs) were 2.6 (interquartile range 2.5, 2.7), 1.9 (1.8, 2.0), 3.2 (3.0, 4.1), and 3.8 (3.4, 4.6), and anti-FXa levels were 0.36 (0.32, 0.38), 0.33 (0.26, 0.39), 0.45 (0.41, 0.51), and 0.62 (0.52, 0.65) U mL-1, respectively. Stage II enrollment was stopped (n = 7) after one serious thrombotic event. Ischemic and bleeding events were rare and, in this small population, showed no clear relation to DX-9065a dose. Elective PCI is feasible using a direct FXa inhibitor for anticoagulation. Predictable plasma drug levels can be rapidly obtained with double-bolus and infusion DX-9065a dosing. Monitoring of DX-9065a may be possible using whole-blood INR. Direct FXa inhibition is a novel and potentially promising approach to anticoagulation during PCI that deserves further study.


Cell cycle-dependent fluctuations in IgE secretion in a human myeloma line.
IgE production was examined in a human myeloma cell line (U266) in cultures in exponential growth and in cultures where cell growth was synchronized by thymidine blockade. Antibody secretion is shown to be restricted to late G1/early S phases of the cell cycle in synchronized cultures, as judged by the slope of the curve relating supernatant IgE levels to time. Employing the ELISA-plaque assay to enumerate cells in active IgE secretion, the frequency of the latter was also shown to peak in S phase. These data collectively suggest that IgE gene transcription occurs only during limited periods of the cell cycle. These results are discussed in terms of using U266 to study the mechanism of action of regulatory T-cell factors which affect IgE synthesis and/or secretion.


Diazonium inactivation in simultaneous-coupling and product inhibition in post-coupling azo-techniques for demonstrating activity of acid hydrolases.
In this communication results are presented of an investigation in which the activity of the hydrolytic enzymes acid phosphatase, beta-glucuronidase, non specific arylesterase, microsomal arylsulphatase, beta-galactosidase, beta-N-acetylglucosaminidase, acid alpha-glucosidase and aminopeptidase M are demonstrated in tissue sections with simultaneous- and post-coupling azo-techniques. Semipermeable membrane techniques are used to hamper enzyme diffusion during the incubation period. From the histochemical and biochemical findings it appeared that an advantage of the post-coupling techniques over the simultaneous-coupling techniques is that inactivation of the enzymes by the coupling reagents is avoided. On the other hand post-coupling techniques are subject to product inhibition. With kinetic inhibition studies it is found that for microsomal arylsulphatase and non-specific arylesterase this product inhibition is non-competitive. This product inhibition may be a problem for histochemical quantitative post-coupling techniques for the determination of acid hydrolase activity.


Delay in Treatment of Neonatal Seizures: A Retrospective Cohort Study.
Neonatal seizures are common and caused by a variety of underlying disorders. There is increasing evidence that neonatal seizures result in further brain damage. To describe the time interval between diagnosis of amplitude-integrated electroencephalography (aEEG)-confirmed seizures and administration of anti-epileptic drugs (AEDs). Single-centre retrospective cohort study, with full-term infants (n = 106) admitted to a level III neonatal intensive care unit between 2012 and 2017 with seizures confirmed on 2-channel aEEG and corresponding raw electroencephalography traces, treated with AEDs. The time interval between the first seizure on the aEEG registration and AED administration was calculated. Factors associated with early treatment were analysed. The median time interval of initiating treatment of aEEG-confirmed seizures was 01:50 h (interquartile range 00:43-4:30 h). Treatment of aEEG-confirmed seizures was initiated <1 h in 34/106 infants (32.1%), between 1 and 2 h in 21/106 infants (19.8%), 2-4 h in 23/106 infants (21.7%), 4-8 h in 14/106 infants (13.2%), and ≥8 h in 14/106 infants (13.2%). Seizures treated <1 h were significantly more often recognized by the seizure detection algorithm (SDA) compared to seizures treated >1 h (67 vs. 42%, p = 0.02) and showed more clinical signs (79.4 vs. 37.5%, p < 0.01). There was no difference for out-of-office hours (23.5 vs. 22.2%, p = 0.88). With only 32.1% of the seizures being treated <1 h, there is room for improvement. Timely treatment occurred more often when seizures were clinical or recognised by the SDA. aEEG is a helpful tool for diagnosing seizures 24/7.


Limited efficacy of antimicrobial metaphylaxis in finishing pigs: A randomized clinical trial.
Pigs that die from pathogens associated with porcine respiratory disease complex (PRDC) in the late finishing period represent a significant economic wastage. While it is common to apply antimicrobial metaphylaxis (AM) to control PRDC, there are few studies exploring the potential cost-saving benefits of AM. In this study we examined the value of using AM in commercially reared, late finishing pigs, from farms with endemic PRDC. A total of 732 pigs from four AIAO wean to market sources, were blocked into 2 matching cohorts, based on enrollment body weight, sex, and rectal temperature. The cohorts received either control (C) or AM (Tulathromycin 2.5mg/kg IM, Zoetis, Florham Park, NJ, USA). Post treatment weight gain over the 21 day period was used as a measure of health and productivity. The AM treated pigs in the lowest weight quartile at enrollment, showed a significantly improved weight gain over controls (18.5 kg vs. 16.4 kg, mean difference=2.1 kg, CI 1.10-3.10, p=0.005) that was not evident in any other starting weight quartiles. These results indicate that the biological advantage and associated improvement in growth efficiency associated with the use of AM against PRDC, is only conferred to a specific sub-set of animals. The economic advantage of this strategy is therefore, only likely if the indicators of potential benefit (e.g., lighter weight cohort) can be reliably established. Further studies are needed to determine whether targeted AM could be effectively applied across the industry.


Glucocorticoid-dependent transformation by human papillomavirus type 16 E7 coding and 3' noncoding sequences.
The establishment of transformation of primary rodent cells by human papillomavirus (HPV) type 16 DNA requires glucocorticoid hormones (Pater et al., Nature 335, 832-835, 1988). Here we provide evidence by mutational analysis that, in the context of the hormone-regulated HPV 16 promoter/enhancer, the only protein coding sequences of HPV 16 required are those of the E7 gene. Moreover, additional sequences adjacent to the 3' end of E7 coding sequences are also essential for the establishment of the transformed phenotype. Splice donor sites, especially an E7 ORF 3' proximal one, are implicated for this cis-acting function, since specific deletion mutations of these splice sites greatly or completely reduced the frequency of transformation and the level of E7 RNA.