Enhancing detection sensitivity in gradient liquid chromatography via post-column refocusing and strong-solvent remobilization.
We developed earlier the post-column refocusing strategy for isocratic separations, which employs trapping target analytes after an analytical separation and additionally focusing them using a strong remobilization solvent prior to detection, and have now extended it to high-speed gradient LC. A gradient separation of antibiotics and its metabolites, applying a linear aqueous acetonitrile gradient from 2 to 65% (v/v) ACN containing 0.1% FA in 10min, performed on an analytical column was selected as an application. Eluted heart-cut fractions were directed from the analytical silica C18 column to a trap column packed with Hypercarb particles. The remobilization of the target analytes was performed in back-flush mode using solvent mixtures tuned to maximize the solvent strength by mixing isopropanol into the remobilization solvent containing acetonitrile. Additionally, a viscosity-calibration experiment showed that the viscosity difference between trapping and remobilization solvents should be smaller than 0.15mPa·s to prevent viscous fingering. To keep the viscosity difference below this limit, during the gradient separation performed on the analytical column, the composition of the remobilization solvent was changed in time. An empirical equation is provided that allows for the selection of the optimal remobilization-solvent composition. To maximize the signal enhancement, the loading time of target analytes on the trap column should be optimized. Peak dispersion was further minimized by applying a flow rate that corresponded to the optimal van-Deemter flow rate of the trap column (20μL/min). Finally, decreasing the diameter of the trap column from 1mm to 0.3mm led to a significant enhancement of the detection sensitivity with more than one order of magnitude. Using an optimized trap configuration and elution/remobilization conditions, a signal enhancement of a factor of 14 was achieved for sulfaguanidine (early-eluting compound in the gradient separation) and 7.3 for furazolidone (late-eluting compound).


Isolation and characterization of yeast nicotinamide adenine dinucleotide kinase.
NAD+ kinase (ATP:NAD+ 2'-phosphotransferase, EC 184.108.40.206) from yeast has been purified utilizing ion-exchange and NAD+-agarose affinity chromatography to give a 2100-fold purification. The apparent homogeneity of the enzyme preparation was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and analytical ultracentrifugation. The enzyme has a subunit molecular weight of 31,000, and a native molecular weight of 124,000, and is, thus, probably a tetramer. The single form of the enzyme has an apparent isoelectric point of 5.85. Initial velocity studies in the forward direction with both substrates gave intersecting Lineweaver-Burk plots, and this suggests a sequential mechanism in which both substrates are bound before products are released. Replots of these data were linear and gave Km values for NAD+ and ATP of 0.68 mM and 2.3 mM, respectively.


A systematic review of case-mix adjustment models for stroke.
To identify any externally validated prognostic model for predicting outcome in unselected populations following acute stroke comprising variables feasible for collection in routine care. Searches were run in MEDLINE, EMBASE, CINAHL, PsycInfo, AMED and ISI Web of Science with no limits on publication date or language. Any study describing the development or external validation of a discernible prognostic model to predict any valid outcome following acute stroke was included. Papers were retained if they met pre-specified inclusion criteria identified from previous reviews and pertinent discussion papers. Data extraction focused on methodological quality of model development, generalizability and feasibility of variable collection. Model performance was examined through consideration of external validation studies. Seventeen externally validated models were identified from 43 papers fulfilling inclusion criteria. Quality of studies describing model development was variable and model performance in external validation studies was generally poor. Models were generally constructed through secondary use of randomized trial or stroke database data. Prognostic variables broadly encompassed markers of stroke severity, pre-stroke function and comorbidities. One model that fulfilled the review criteria and had extensive external validation in a range of post-stroke populations was identified (the Six Simple Variables model). The Six Simple Variables model performed well in six external validation studies, although prediction of outcome in patients with milder strokes was less reliable. Other models identified in this review have been developed using robust methodology but comprise more complex clinical variables which may limit their utility in routine stroke care.


Comparison of caprine mycoplasmosis caused by Mycoplasma capricolum, Mycoplasma mycoides subsp. mycoides, and Mycoplasma putrefaciens.
We reviewed the disease process in goats caused by Mycoplasma capricolum, M. mycoides subsp. mycoides, and M. putrefaciens and compared mycoplasma-infected goats (particularly young kids) for microanatomic changes. M. capricolum and M. mycoides subsp. mycoides cause a nearly identical disease: septicemia leading to pyrexia, high morbidity and mortality, interstitial pneumonia, arthritis (including polyarthritis), and mastitis. Until recently, M. putrefaciens was known only as a cause of mastitis. However, a new isolate of M. putrefaciens may be transiently septicemic, as evidenced by polyarthritis with isolations from joints, lung, brain, kidney, pleural fluid, uterus, and urine.


Yukmijihwang-tang ameliorates ischemia/reperfusion-induced renal injury in rats.
The present study was designed to examine whether Yukmijihwang-tang (YJT), which is a Korean decoction for the treatment of renal disease, has an effect on renal functional parameters in association with the expression of aquaporin 2 (AQP 2), Na,K-ATPase, heme oxygenase-1 (HO-1) in rats with ischemia/reperfusion-induced acute renal failure (ARF). Polyuria caused by down-regulation of renal AQP 2 in the ischemia/reperfusion-induced ARF rats was markedly restored by administration of YJT (100 or 200 mg/kg, p.o.) with restoring expression of AQP 2 in the kidney. The expressions of Na,K-ATPase alpha1 and beta1 subunits in the renal medulla and cortex of the ARF rats were also restored in them by the administration of YJT. Administration of YJT lowered the expression of renal HO-1, which was up-regulated in rats with ischemia/reperfusion-induced ARF. The renal functional parameters including creatinine clearance, urinary sodium excretion, urinary osmolality, and solute-free reabsorption were also markedly restored in ischemia-ARF rats by administration of YJT. Histological study also showed that renal damages in the ARF rats were abrogated by administration of YJT. Taken together, these data indicate that YJT ameliorates renal defects in rats with ischemia/reperfusion-induced ARF.


Animal-derived products in science and current alternatives.
More than fifty years after the 3Rs definition and despite the continuous implementation of regulatory measures, animals continue to be widely used in basic research. Their use comprises not only in vivo experiments with animal models, but also the production of a variety of supplements and products of animal origin for cell and tissue culture, cell-based assays, and therapeutics. The animal-derived products most used in basic research are fetal bovine serum (FBS), extracellular matrix proteins such as Matrigel™, and antibodies. However, their production raises several ethical issues regarding animal welfare. Additionally, their biological origin is associated with a high risk of contamination, resulting, frequently, in poor scientific data for clinical translation. These issues support the search for new animal-free products able to replace FBS, Matrigel™, and antibodies in basic research. In addition, in silico methodologies play an important role in the reduction of animal use in research by refining the data previously to in vitro and in vivo experiments. In this review, we depicted the current available animal-free alternatives in in vitro research.


Lung Cancer Screening Among Mammography Patients: Knowledge, Eligibility, Participation, and Interest.
To determine lung cancer screening eligibility, knowledge, and interest and to quantify the effect of the expanded 2021 lung cancer screening eligibility criteria among women presenting for screening mammography, a group with demonstrable interest in cancer screening. A single-page survey was distributed to patients presenting for screening mammography, from January-March 2020 and June 2020-January 2021, at 2 academic medical centers on the East and West Coasts. The population served by the East Coast institution has greater poverty, greater ethnic/racial diversity, and lower education levels. Survey questions included age, smoking history, lung cancer screening knowledge, participation, and interest. Lung cancer screening eligibility was determined for both 2013 and 2021 USPSTF guidelines. Descriptive statistics were calculated, and data were compared between groups using the Chi-square test, Mann-Whitney nonparametric test, and the 2-sample t test. 5512 surveys were completed; 33% (1824) of women reported a history of smoking-30% (1656) former smokers and 3% (156) current smokers. Among women with a smoking history, 7% (127/1824) were eligible for lung cancer screening using 2013% and 11% (207/1824) using the 2021 USPSTF criteria. Interest in lung cancer screening was high (73%; 151/207) among eligible women using 2021 USPSTF criteria, but only 42% (87/207) had heard of lung cancer screening and only 28% (57/207) had received prior LDCT screening. Eligible screening mammography patients reported high levels of interest in lung cancer screening but low levels of knowledge and participation. Linking mammography and LDCT appointments may improve lung cancer screening participation.


Proabsorptive and prosecretory roles for nitric oxide in cholera toxin induced secretion.
Cholera toxin causes small intestinal hypersecretion by inducing a coordinated response from enterocytes, enterochromaffin cells, enteric neurones, and the vascular supply. Nitric oxide has been implicated in the function of these separate components. To explore the role of nitric oxide in the totality of cholera toxin induced secretion in vivo. One group of adult male Wistar rats was treated with the nitric oxide synthase inhibitors NG-nitro-L-arginine methyl ester (L-NAME; subcutaneously or intraluminally), NG-methyl-L-arginine (L-NMA), or 7-nitroindazole. A second group of rats was treated with L-arginine (intraperitoneally or intraluminally) or D-arginine. The small intestine was isolated between two cannulae and instilled with 75 microg cholera toxin or saline for two hours. Small intestinal perfusion of a plasma electrolyte solution containing [14C]-PEG was undertaken to determine net water and electrolyte movement. After the experiment macroscopic and microscopic intestinal appearances were noted and jejunal 5-hydroxytryptamine concentrations were determined. Both L-arginine and L-NAME induced secretion in the basal state, but only when administered intraluminally. Systemically applied L-NAME caused a dose dependent reduction in cholera toxin induced secretion. This was paralleled by L-NMA but not by 7-nitroindazole or by intraluminally applied L-NAME. Systemically applied L-NAME caused notable cyanosis of the intestine, consistent with mesenteric ischaemia, but no microscopic abnormalities. Systemically applied L-arginine but not D-arginine also reduced cholera toxin induced secretion and inhibited 5-hydroxytryptamine release. Nitric oxide has a duality of roles in cholera toxin induced secretion, acting both as an absorbagogue and a secretagogue. Its mechanisms of action include the maintenance of mucosal perfusion and enterochromaffin cell stabilisation.


Quantification of minerals from ATR-FTIR spectra with spectral interferences using the MRC method.
A method for quantifying the individual components of mineral samples based on attenuated total reflectance - Fourier transform infrared spectroscopy (ATR-FTIR) is described, extending the constant ratio (CR) method to analytes absorbing in a common range of wavenumbers. Absorbance values in the spectral region where the analytes absorb relative to the absorbance of an internal standard absorbing at a wavenumber where the analytes do not absorb, permits the quantification of N analytes using measurements at N fixed wavenumbers. The method was tested for mixtures of albite, orthoclase, kaolin and quartz.


Evolution of the mitochondrial genetic code. III. Reassignment of CUN codons from leucine to threonine during evolution of yeast mitochondria.
Yeast mitochondria use UUR as the sole leucine codons. CUN, universal leucine codons, are read as threonine by aberrant threonine tRNA with anticodon sequence (UAG). The reassignment of CUN codons to threonine during yeast mitochondrial evolution could have proceeded by the disappearance of CUN codons from the reading frames of messenger RNA, through mutation mainly to UUR leucine codons as a result of AT pressure. We suggest that this was accompanied by a loss of leucine-accepting ability of tRNA Leu(UAG). This tRNA could have then acquired threonine-accepting activity through the appearance of an additional threonyl-tRNA synthetase. CUN codons that subsequently appeared from mutations of various other codons would have been translated as threonine. This change in the yeast mitochondrial genetic code is likely to have evolved through a series of nondisruptive nucleotide substitutions that produced no widespread replacement of leucine by threonine in proteins as a consequence.


Purification to homogeneity of a high molecular weight human B cell growth factor; demonstration of specific binding to activated B cells; and development of a monoclonal antibody to the factor.
High molecular weight B cell growth factor (HMW-BCGF) produced by a T cell line was purified to homogeneity and demonstrated to bind specifically to activated human B cells. A monoclonal antibody to HMW-BCGF was developed that (a) specifically inhibited the activity of HMW-BCGF in enhancing B cell proliferation, (b) specifically bound to HMW-BCGF in Western blots, (c) specifically absorbed HMW-BCGF activity from culture supernatants, and (d) specifically absorbed an internally labeled protein from T-ALL supernatant which comigrates with HMW-BCGF on sodium dodecyl sulfate-polyacrylamide gels. This antibody should help in cloning the gene for HMW-BCGF and further exploring the physiologic roles of HMW-BCGF.


Pharmacokinetics, Efficacy, and Safety of a SARS-CoV-2 Antibody Treatment in Pediatric Participants: An Open-Label Addendum of a Placebo-Controlled, Randomized Phase 2/3 Trial.
Bamlanivimab and etesevimab (BAM + ETE) are monoclonal antibodies (mAbs) effective in reducing COVID-19-related hospitalizations and all-cause mortality in adult participants at increased risk for severe disease. We present pharmacokinetic (PK), efficacy, and safety results from pediatric participants (< 18 years of age) with COVID-19 who were treated with BAM + ETE. In an addendum to the phase 2/3 BLAZE-1 clinical trial (NCT04427501), pediatric participants received open-label weight-based dosing (WBD, n = 94) based on exposure-matching to the authorized dose of BAM + ETE in adult participants. For efficacy and safety assessments, placebo (n = 14) and BAM + ETE (n = 20)-treated adolescent participants (> 12 to < 18 years of age) from the BLAZE-1 trial were included in the overall pediatric population (N = 128). All participants had mild to moderate COVID-19 upon enrollment and ≥ 1 risk factor for severe COVID-19. The primary objective was to characterize the PK of BAM and ETE in the WBD population. The median age of the participants was 11.2 years, 46.1% were female, 57.9% were Black/African American, and 19.7% were Hispanic/Latino. The area under the curve for BAM and ETE in the WBD population was similar to that previously observed in adults. There were no COVID-19-related hospitalizations or deaths. All adverse events (AE) except one were mild or moderate, with one participant reporting a serious AE. WBD in pediatric participants achieved similar drug exposures compared to adult participants that received the authorized BAM + ETE dose. The pediatric efficacy and safety data were consistent with adults receiving mAbs for COVID-19. NCT04427501.


Clinicocytopathological spectrum, including uncommon forms, of nine cases of chordomas with immunohistochemical results, including brachyury immunostaining: A single institutional experience.
To present clinical and cytopathological features of nine cases of chordomas, diagnosed over 9 years and confirmed by brachyury (T) immunostaining. Conventional cytological smears, stained with Papanicolaou and May-Grünwald Giemsa, along with corresponding histopathological (n = 8) and immunostained sections (n = 8) were reviewed. Immunohistochemical staining was performed on tissue sections by polymer detection technique. Nine tumours occurred in seven males and two females, with age ranging from 36 to 72 years (average = 58.7), in the sacrum (seven) and spine (two). On fine needle aspiration cytology, five cases were either diagnosed with or diagnosed with a suggestion of a chordoma, while three cases were diagnosed with chordoma as a differential diagnosis. On review, smears were moderately cellular, comprising myxoid stroma (9/9), epithelioid cells (9/9), physaliphorous cells (8/9), including binucleation (7/9), prominent nucleolisation (2/9), pleomorphic cells (2/9) and intranuclear inclusions (3/9). Immunohistochemically, tumour cells expressed cytokeratin (4/4), pan cytokeratin (4/4), epithelial membrane antigen (8/8), S100 protein (6/8) and brachyury (8/8). Five patients underwent surgical excision, including two who underwent adjuvant radiotherapy (RT) and four patients who underwent RT. During follow-up (n = 8), a single patient developed recurrence and another presented with metastatic lesions. Finally, five patients were alive with disease (7-53 months); a single patient was free of disease (4 months), and two patients died of disease; the latter cases displayed pleomorphic cells and intranuclear inclusions. Chordomas can be primarily diagnosed by fine needle aspiration cytology in a typical clinicoradiological setting with a combination of key cytomorphological features. Pleomorphic cells and intranuclear inclusions are associated with a relatively aggressive subtype. An exact diagnosis has treatment implications and requires confirmation by brachyury immunostaining.


Comparison of Trypanosoma cruzi detection by PCR in blood and dejections of Triatoma infestans fed on patients with chronic Chagas disease.
In this study, we compare the sensitivity of detecting Trypanosoma cruzi in dejections of Triatoma infestans nymphs that had fed on the blood of chronic chagasic patients, with detection of T. cruzi in peripheral blood, using a polymerase chain reaction assay (PCR-D and PCR-B, respectively). Fifty-seven chronic patients were evaluated who were positive (group I) or negative (group II) by xenodiagnosis (XD). Patients showed 84.8 and 75% positive PCR results in both kinds of samples in groups I and II, respectively. Six cases (10.5%) showed positive PCR-D and negative PCR-B, five of them belonged to group I. In contrast, five cases of group II showed negative PCR-D and positive PCR-B. Overall, the PCR-D assay gave positive results in 52 out of 57 samples (91.2%), while 51 out of 57 (89.5%) were positive by PCR-B. In comparison, only 57.9% were positive by XD (p = 0.0001). In conclusion, PCR performed in dejection or blood was more sensitive for the parasite detection than xenodiagnosis. All patients (100%) were detected positive when both, PCR-D and PCR-B, were applied.


Effect of high-pressure processing pretreatment on the physical properties and colour assessment of frozen European hake (Merluccius merluccius) during long term storage.
Fish freshness is lost by autolytic degradation produced by endogenous enzymes. Frozen storage is one of the most used methods to preserve fish properties. However, protein denaturation has shown to be a major problem for frozen European hake (Merluccius merluccius), leading to texture losses and off-odour development. The aim of this work was to study the changes produced by high-pressure processing (HPP) before freezing on quality of frozen European hake stored at -21 °C for 12 months. The effect of HPP (150-450 MPa) on mechanical properties and expresible water was evaluated in raw and cooked fish samples. The effect on colour (L*, a* and b*) was assessed only in raw fish. Results showed that HPP before freezing is beneficial to maintain expresible water in good levels up to 6 months. The luminosity significantly increased with pressure level. Textural profile of raw samples showed that HPP increased hardness, adhesiveness and springiness of frozen hake. Cooked samples were also affected by HPP, being the best results obtained at 300 MPa for 6 months of frozen storage. Overall, results showed that HPP improves the quality of frozen hake.


Cardiorespiratory demands of competitive rock climbing.
Rock climbing has become a mainstream sport, contested on the Olympic stage. The work/rest pattern of bouldering is unique among disciplines, and little is known about its physiological demands. This study characterised the cardiorespiratory responses to simulated competition. Eleven elite boulderers (7 male) volunteered to participate (age = 23.3 ± 4.5 years; mass = 68.2 ± 9.7 kg; stature = 1.73 ± 0.06 m; body fat = 10.4% ± 5%). Subjects completed incremental treadmill running to determine maximal capacities. On a separate day, they undertook a simulated Olympic-style climbing competition comprising 5 boulder problems, each separated by 5 min of rest. Pulmonary ventilation, gas exchange, and heart rate were assessed throughout. Total climbing time was 18.9 ± 2.7 min. Bouldering elicited a peak oxygen uptake of 35.8 ± 7.3 mL·kg-1·min-1 (∼75% of treadmill maximum) and a peak heart rate of 162 ± 14 beats·min-1 (∼88% of maximum). Subjects spent 22.9% ± 8.6% of climbing time above the gas exchange threshold. At exercise cessation, there was an abrupt and significant increase in tidal volume (1.4 ± 0.4 vs. 1.8 ± 0.4 L; p = 0.006, d = 0.83) despite unchanged minute ventilation. Cardiorespiratory parameters returned to baseline within 4 min of the rest period. In conclusion, competitive bouldering elicits substantial cardiorespiratory demand and evidence of tidal volume constraint. Further studies are warranted to explore the effect of cardiorespiratory training on climbing performance. Novelty: Competitive bouldering evokes a high fraction of maximal oxygen uptake and prolonged periods above the gas exchange threshold. Climbing appears to impose a constraint on tidal volume expansion. Cardiorespiratory indices in elite climbers return to baseline within 2-4 min.


Online surveys for BGLT research: issues and techniques.
Online surveys are becoming increasingly popular for accessing less visible and decentralized populations, including bisexual, gay, lesbian and transgender (BGLT) populations. Advances in technology and convenience for the both the researcher and the participant have facilitated this trend. In this paper, we explore issues related to conducting BGLT survey research online, such as making decisions about survey formats, target populations and recruitment, compensation, access.


Reconstruction of Intrapelvic Defects Using the Free Anterolateral Thigh Flap: Expanding the Traditional Algorithm.
Reconstruction of intrapelvic soft tissue defects traditionally relies on regional pedicled myocutaneous flaps. However, there remain situations in which local options are unavailable. We review our experience treating intrapelvic defects with the anterolateral thigh (ALT) microvascular free flap. A retrospective, institutional review was conducted from 2014 to 2018 of patients undergoing microvascular ALT flap reconstruction of intrapelvic defects. Four patients were identified in this cohort out of 92 total pelvic reconstruction cases. All patients underwent abdominoperineal resection (APR) for rectal cancer treatment. In the two male patients, pelvic abscesses and bladder leak necessitated soft tissue reconstruction after radiation and APR. In both, regional tissue options were unavailable, and a buried ALT free flap was used for soft tissue reconstruction. Both female patients developed rectovaginal fistulas secondary to their tumor burden, necessitating posterior vaginal wall resections. Prior surgical scars and ostomies made abdominal wall tissue unavailable; thus, free ALTs were used to eliminate intrapelvic dead space and reconstruct the posterior vaginal wall. In all cases, recipient vessels were the deep inferior epigastric artery and vein. Flap survival was 100%. Pelvic reconstruction has traditionally been addressed with local/regional pedicled flaps. In cases where these are unavailable, the free ALT flap is a versatile option when buried for intrapelvic reconstruction or posterior vaginal wall lining. We also propose updating Cordeiro et al.'s classic vaginal defect reconstruction algorithm to include the free ALT flap for type IB cases in which the rectus abdominis is unavailable.


Freeze fracture and numerical analyses of the spiral ganglion cells.
The freeze-fracture technique was used to study the spiral ganglion cell population in the adult mouse. The majority of the ganglion cells (type I) had a smooth cell surface. Interspersed among these first ganglion cells, a small number of other ganglion cells (type II) showed considerable membrane specializations and were morphologically reminiscent of nerve terminals previously described only in human temporal bones. Calculations of the number of the two types of spiral ganglion cells in sectioned material showed that the adult mouse spiral ganglion comprises of 3%-4% unmyelinated type II ganglion cells.


The impact on clinical outcomes of post-operative radiation therapy delay after neoadjuvant chemotherapy in patients with breast cancer: A multicentric international study.
Radiation therapy (RT) is frequently used for post-operative treatment in breast cancer (BC) patients who received preoperative systemic therapy (PST) and surgery. Nevertheless, the optimal timing to start RT is unclear. Data from BC patients who underwent chemotherapy as PST, breast surgery and RT at 3 Institutions in Brazil and Canada from 2008 to 2014 were evaluated. Patients were classified into three groups regarding to the time to initiation of RT after surgery: <8 weeks, 8-16 weeks and >16 weeks. A total of 1029 women were included, most of them (59.1%; N = 608) had clinical stage III. One hundred and forty-one patients initiated RT within 8 weeks, 663 between 8 and 16 weeks and 225 beyond 16 weeks from surgery. With a median follow-up of 32 months, no differences in disease-free survival (DFS), overall survival and locoregional recurrence-free survival (LRRFS) were observed of time to indicated RT (<8 weeks versus 8-16 weeks versus >16 weeks). However, in luminal subtype patients (46.5%; N = 478), initiation of RT up to 8 weeks after surgery was associated with better LRRFS (<8 weeks versus >16 weeks: HR 0.22; 95%CI 0.05-0.86; p = 0.03), with a tendency to a better DFS (<8 weeks versus >16 weeks: HR 0.50; 95%CI 0.25-1.00). RT initiated up to 8 weeks after surgery was related to better LRRFS in luminal BC patients who underwent PST. Our results suggest that early start of RT is important for these patients.


Estimating the volume of alcohol-based hand rub required for a hand hygiene program.
Providing alcohol-based hand rub (ABHR) at the point of care is a key success factor in enabling health care providers to achieve optimal hand hygiene practices. There are few tools available for health care organizations to assess the number of points of care, estimate the number of hand hygiene indications at each point of care, and estimate the anticipated volume of ABHR required to support a hand hygiene program. We developed an assessment tool to systematically evaluate the environmental hand hygiene needs in diverse care settings across a multisite health care organization. We identified 1,103 points of care in 34 clinical units, of which only 53% had ABHR at point of care. There are an estimated 171,468,240 (95% confidence interval: 146,844,406-191,871,179) hand hygiene indications per year in our in-patient and emergency areas. If 100% compliance with hand hygiene is achieved, 240,056 L of ABHR will be required each year. Our environmental assessment was invaluable in estimating the number of hand hygiene indications by unit and the logistical and financial requirements to implement a hand hygiene program. Other health care organizations may find this a useful framework to estimate their own environmental hand hygiene needs.


Subchronic lead feeding study in male rats and micropigs.
This study compared the lead uptake from contaminated test soil of known lead concentration with a soluble lead acetate standard, which was considered to be 100% bioavailable. This study also compared the lead bioavailability from this lead-contaminated soil between rats and micropigs. Harlan Sprague-Dawley rats and Yucatan micropigs were fed lead-contaminated soil as a 5% (w/w) mixture with their diet. The lead-contaminated soil was either a specific test soil of known lead concentration (1000 microg/g) or basal low concentration lead soil ( approximately 135 microg/g), which was spiked with lead acetate to match the lead content of the test soil. The effective diet lead concentration was 50 microg Pb/g diet. Results demonstrated that rats reached steady-state concentrations of blood lead by week 2, whereas micropigs did not reach steady state until week 4. In addition, final blood lead concentrations in micropigs were four-fold higher than those in rats. In the micropigs, the final blood lead levels in the test soil study group were significantly lower than those in the lead acetate study group, although there was no significant difference between study groups in the rats. Tissue lead concentrations were significantly higher in micropigs than those in the rats, although the diet lead concentrations in both sets of animals were the same. In summary, because of the greater sensitivity in demonstrating higher tissue lead incorporation in micropigs as compared to rats, the micropig is a better animal model for demonstrating the differences in relative lead bioavailability when testing different sources of lead-contaminated soils.


Serial SOFA-score trends in ICU-admitted COVID-19 patients as predictor of 28-day mortality: A prospective cohort study.
The efficacy of Sequential Organ Failure Assessment (SOFA) score as predictor of clinical outcomes among ICU-admitted COVID-19 patients is still controversial. We aimed to assess whether SOFA-score in different time intervals could predict 28-day mortality compared with other well-acknowledged risk factors of COVID-19 mortality. This observational prospective cohort was conducted on 1057 patients from March 2020 to March 2022 at Masih Daneshvari Hospital, Iran. The univariate and multivariate Cox proportional analysis were performed to assess the hazards of SOFA-score models. Receiver operating characteristic (ROC) curves were designed to estimate the predictive values. Mean SOFA-score during first 96 h (HR: 3.82 [CI: 2.75-5.31]), highest SOFA-score (HR: 2.70 [CI: 1.93-3.78]), and initial SOFA-score (HR: 1.65 [CI: 1.30-2.11]) had strongest association with 28-day mortality (p < .0001). In contrast, SOFA scores at 48 and 96 h as well as Δ-SOFA: 48-0 h and Δ-SOFA: 96-0 h did not show significant correlations. Among them, merely mean SOFA-score (HR: 2.28 [CI: 2.21-3.51]; p < .001) remained as independent prognosticator on multivariate regression analysis; though having less odds of predicting value compared with age (HR: 3.81 [CI: 1.98-5.21]), hypertension (HR: 3.11 [CI: 1.26-3.81]), coronary artery disease [CAD] (HR: 2.82 [CI: 1.51-4.8]), and diabetes mellitus (HR: 2.45 [CI: 1.36-2.99]). The area under ROC (AUROC) for mean SOFA-score (0.77) and highest SOFA-score (0.71) were larger than other SOFA intervals. Calculating the first 96 h of SOFA trends, it was obtained that fatality rate was <12.3% if the score dropped, between 28.8% and 46.29% if the score remained unchanged, and >50.45% if the score increased. To predict the 28-day mortality among ICU-admitted COVID-19 patients, mean SOFA upon first 96 h of ICU stay is reliable; while having inadequate accuracy comparing with well-acknowledged COVID-19 mortality predictors (age, diabetes mellitus, hypertension, CAD). Notably, increased SOFA levels in the course of first 96 h of ICU-admission, prognosticate at least 50% fatality regardless of initial SOFA score.


A Systematic Review of the Efficacy of Contingency Management for Substance Use Disorders in Low and Middle Income Countries.
The impact of illicit substance use is especially devastating in low-resourced countries where factors such as poverty, unemployment, and inadequate services impede successful treatment. Contingency management (CM) is a treatment for substance use disorders that has shown to be effective in eliciting behaviour change. The efficacy of CM interventions in low and middle income countries (LMICs) has been under explored. The aim of this systematic review of randomized controlled trials was to assess measures of CM efficacy in addressing substance use disorders, while also considering contextual moderators of CM in LMICs. A search of PubMed, Scopus, and Cochrane library databases yielded 18 studies for inclusion, from which relevant data were extracted using modified versions of the Cochrane Characteristics of Studies tool. Two studies were located in a low-income country, two in lower-middle income countries, and fourteen in upper middle-income countries. Overall, estimated efficacy estimates were similar to those from higher income countries. However, context-specific challenges that warrant further investigation included limited access to trained staff and structural and financial constraints. While CM in LMICs is in its early stages of development, efficacy estimates were not substantially different compared to high income countries. Challenges such as costs, willingness to implement, and the stigma associated with addiction sets the stage for further research in these contexts.


Modification of the ferrozine technique to analyze iron contents in different foods: comparative study using an internal standard as reference methodology.
A methodology for the determination of iron in foods fortified with this element or in nutritional products is important and has to be sensitive and rapid. In developing countries, an inexpensive and reliable methodology is also required. For this purpose, the Gordon's Ferrozine technique was slightly modified and assayed with yogurt, dry powdered milk, and cereal mixtures, all of them fortified with iron, using an internal standard as the reference methodology. The obtained results demonstrate a close correlation between the standard curve interpolation method and the internal standard reference method (correlation coefficient r2 = 0.9950) in a wide range of concentrations. The slope (0.9998+/-0.0040) demonstrates that both procedures measure equal amounts of iron. The conclusion is that the proposed technique is a reliable, practical, and inexpensive methodology for iron determination in different foods fortified with iron.


Acute lateral dislocation of the patella: correlation of ultrasound scanning with operative findings.
Acute lateral dislocation of the patella has been associated with disruption of the medial restraints of the patella. Following non-operative management there is a re-dislocation rate of up to 44%. The purpose of this study was to test whether sonography is a reliable method of assessing the medial retinaculum after acute dislocation of the patella. Ten patients following acute patellar dislocation had an ultrasound scan (USS) performed by an experienced musculoskeletal radiologist. Each patient subsequently had an examination under anaesthetic, arthroscopy, and repair of the ruptured structures. The ultrasound reports were compared to the surgical findings to determine the accuracy of this investigation. USS located deficiencies in the ligamentous attachments to the medial border of the patella and the presence of avulsed bony fragments, all of which were confirmed at operation. The sonographic diagnosis of haematoma or torn fibres in the vastus medialis obliquus (VMO) corresponded with our operative findings. The most significant findings were the correlation of free fluid around the medial collateral ligament (MCL) with avulsion of the femoral attachment of the medial patellofemoral ligament (MPFL) and the presence of avulsed fragments of bone from the medial border of the patella.


Precipitation reactions with Newcastle disease virus.
Acid precipitated and detergent treated Newcastle disease virus (NDV) antigen was prepared and characterised using the agar gel precipitation (AGP) test. The detergent treated NDV antigen was used to screen antibodies to NDV and the results compared with the conventional haemagglutination inhibition (HI) test titres. AGP test could detect NDV antibodies in serum samples when the corresponding HI titres were 1:8 and above. Detergent treatment of Newcastle disease virus greatly reduced its haemagglutinating ability. Simultaneous detection of antibodies to NDV and infectious bursal disease virus in AGP test was attempted and found successful.


Palliative Care and Symptom Management in Older Patients with Cancer.
Older patients with cancer are best served by a multidisciplinary approach with palliative care (PC) playing an integral role. PC focuses on symptom control irrespective of its cause and should not be associated only with terminal care. It provides an additional layer of support in the care of patients with cancer with an emphasis on quality of life. This article discusses the evaluation and management of pain and other common nonpain symptoms that occur in elderly patients with cancer, as well as end-of-life care.


Is the use of vestibular information weighted differently across the initiation of walking?
The purpose of this experiment was to examine vestibular contributions at specific times during the initiation of walking in human subjects. Subjects began walking forward at the sound of an auditory tone, with vision present or occluded. Galvanic vestibular stimulation (GVS) was delivered with the anode electrode on the right or left side at either: (1) onset of the anticipatory postural adjustment (APA), (2) toe-off of the first swing limb (TO) or (3) heel contact of the first swing limb (HC). Ground reaction forces and kinematic data were collected. Upper body (roll angles from head, trunk and pelvis) and lower body (foot placement) data were analysed to determine whether the timing and magnitude of the response to GVS, and therefore the level of vestibular contribution, was modulated at different points during the initiation of gait. With vision present and occluded, the magnitude of the lower body response varied depending on the event in the gait cycle at which the stimulation was delivered. These novel results demonstrate evidence that vestibular weighting during gait initiation is dependent upon the specific gait initiation events. Upper body roll also exhibited magnitude differences between events. However, these changes are proposed to occur due to the transition from a stationary position into a dynamic state, prompting the increased weighting of vestibular information. With vision present no significant changes were seen in the segment roll response. The observations suggest a distinction in vestibular regulation of upper body roll versus foot placement for successful completion of the gait initiation task. Changes in upper body roll are influenced by the dynamic nature of the task, whereas foot placement changes are modulated based on the event during gait initiation at which GVS is delivered.


Transcriptome-wide analysis of introgression-resistant regions reveals genetic divergence genes under positive selection in Populus trichocarpa.
Comparing gene expression patterns and genetic polymorphisms between populations is of central importance for understanding the origin and maintenance of biodiversity. Based on population-specific gene expression levels and allele frequency differences, we sought to identify population divergence (PD) genes across the introgression-resistant genomic regions of Populus trichocarpa. Genes containing highly diverged loci [i.e., genetic divergence (GD)] or showing expression divergence (ED) between populations were widely distributed in the genome and substantially enriched in functional categories related to stress responses, disease resistance, timing of flowering, cell cycle regulation, plant growth, and development. Nine genomic regions showing evidence of strong positive selection were overlapped with GD genes, which had significant differences between Oregon (a southernmost peripheral deme) and the other demes. However, we did not find evidence that genes under positive selection show an enrichment for ED. PD genes and genes under selection pertained to the same gene classes, such as SERINE/CYSTEINE PROTEASE, ABC TRANSPORTER, GLYCOSYLTRANSFERASE and other transferases. Our analysis also revealed that GD genes were polymorphic within the species (41.9 ± 3.66 biallelic variants per gene), as previously reported in herbaceous plants. By contrast, ED genes contained less genetic variants (10.73 ± 1.14) and were likely highly expressed. In addition, we found that trans- rather than cis-acting variants considerably contribute to the evolution of >90% PD genes. Overall, this study elucidates that cohorts of PD genes agree with the general attributes of known speciation genes and GD genes will provide substrates for positive selection to operate on.


How to become neural crest: from segregation to delamination.
The development of the neural crest up to the stage where they leave the neural tube can be observed as a series of concatenated but independent events that involve dorsalization of the neural plate/neural tube, neural crest induction, segregation and stabilization, epithelial to mesenchymal transition and delamination. During all these processes, the nascent neural crest cells are subjected to the influence of different signals and have to overcome competition for cell fate and apoptotic signals. In addition, striking rostrocaudal differences unveil how the regulatory cascades are somehow different but still can lead to the production of bona fide neural crest cells.


TRA2: The dominant power of alternative splicing in tumors.
The dysregulation of alternative splicing (AS) is frequently found in cancer and considered as key markers for cancer progression and therapy. Transformer 2 (TRA2), a nuclear RNA binding protein, consists of transformer 2 alpha homolog (TRA2A) and transformer 2 beta homolog (TRA2B), and plays a role in the regulation of pre-mRNA splicing. Growing evidence has been provided that TRA2A and TRA2B are dysregulated in several types of tumors, and participate in the regulation of proliferation, migration, invasion, and chemotherapy resistance in cancer cells through alteration of AS of cancer-related genes. In this review, we highlight the role of TRA2 in tumorigenesis and metastasis, and discuss potential molecular mechanisms how TRA2 influences tumorigenesis and metastasis via controlling AS of pre-mRNA. We propose that TRA2Ais a novel biomarker and therapeutic target for cancer progression and therapy.


[Early treatment principles of tooth replacement disorders associated with hereditary oral diseases].
Tooth replacement disorders are characterized by retention of deciduous teeth and abnormalities in permanent teeth eruption. Hereditary disorders with multiple teeth involved include cleidocranial dysplasia, osteopetrosis and Gardner syndrome. These rare diseases have great difficulty in treatment with various principles reported. This article focused on clinical manifestations and early treatment principles of these hereditary disorders, as well as the important role of dentists in early diagnosis of these diseases.


Chromosomal aberrations induced by chemotherapy and radiotherapy in lymphocytes from patients with breast carcinoma.
Stable chromosomal aberrations (SCAs) have been found in circulating lymphocytes from patients treated for breast carcinoma. Therefore, we tried to define their incidence in such patients, to determine an in vitro dose-effect relationship, and to correlate these data with clinical parameters. This prospective study included 25 patients who, after surgery, underwent either radiotherapy (RT) alone (n = 15) or RT combined with chemotherapy (n = 10). SCAs were scored using the fluorescent in situ hybridization technique before RT and 4 and 12 months after RT. Dose-effect curves were established by in vitro irradiation of blood samples with 2 and 4 Gy, before and after treatment. In all patients, the rate of SCAs increased significantly after external irradiation. No significant decrease in SCAs was observed during the first year after RT. RT and chemotherapy had no effect on the lymphocyte in vitro dose-effect relationship. No relationship was found in the distribution of patients between the yield of SCAs scored after external irradiation and after in vitro irradiation. SCAs after RT or in vitro irradiation did not correlate with family history of breast carcinoma or acute toxicity of treatment. More significantly, the yield of SCA after external irradiation was strongly related to the irradiation of the internal mammary chain and the supraclavicular lymph node area, suggesting that the volume of irradiated blood vessels was an essential parameter in determining the rate of SCAs. A high and stable yield of SCAs persisted at least 1 year after external irradiation. The nature of the volume irradiated containing large blood vessels was the major determinant of the observed biologic dose.


Localization of opiate receptors in substantia nigra evidence by lesion studies.
From the effects of selective lesions it is concluded that opiate receptors are located on dopamine (DA) cell-bodies and/or dendrites as well as on GABA nerve-terminals in substantia nigra.


Prognostic effect of estimated glomerular filtration rate in patients with cardiogenic shock or cardiac arrest undergoing percutaneous veno-arterial extracorporeal membrane oxygenation.
Veno-arterial extracorporeal membrane oxygenation (VA-ECMO) can improve survival in patients with cardiogenic shock or cardiac arrest. We investigated the association between initial renal function and clinical outcome in patients undergoing VA-ECMO for cardiogenic shock and cardiac arrest. This was a single-center, retrospective cohort study of 287 patients who underwent ECMO at our hospital from January 2005 to December 2014. We excluded 70 patients with non-cardiogenic events. The remaining 217 patients were divided into 2 groups according to initial estimated glomerular filtration rate (eGFR): Initial high eGFR (non-renal failure: non-RF) group: eGFR≥60ml/min/1.73m2 (n=73) and initial low eGFR (RF) group: eGFR<60ml/min/1.73m2 (n=144). Clinical outcome was defined as all-cause death at 30 days after extracorporeal life support. VA-ECMO was begun in 87% of patients for cardiac arrest. The non-RF group was significantly younger (51.6 vs. 62.6 years), had lower body mass index (22.8 vs. 24.7kg/m2), lower blood urea nitrogen (14.4 vs. 23.9mg/dl), and lower K (4.0 vs. 4.5mEq/l, all p<0.05) than the RF group. Incidence of all-cause death at 30 days was significantly lower in the non-RF than RF group (49% vs. 76%, p<0.0001). Initial low eGFR was an independent predictor of mortality after adjustment for multiple cofounders (OR: 4.08, 95% CI: 1.77-9.42, p<0.001). Kaplan-Meier curve showed better outcome in the non-RF versus RF group (p=0.0009). An initial low eGFR may predict worse clinical outcome in patients undergoing VA-ECMO for cardiogenic shock and cardiac arrest.


Differential expression within a family of novel wound-induced genes in potato.
Wounding in higher plants leads to an increased synthesis of specific messenger RNAs. A cDNA clone complementary to a wound-induced message from potato tubers was used to isolate a lambda clone from a genomic library of Solanum tuberosum var. Maris Piper. DNA sequence analysis has shown that this single genomic clone contains two novel wound-induced genes, called win1 and win2, organised in close tandem array. The coding sequences of these two genes are highly homologous and are interrupted by a single intron. However, the sequences of the introns and flanking regions have diverged widely. Win1 and win2 encode cysteine-rich proteins of 200 and 211 amino-acids, respectively, which show striking homologies to several chitin-binding proteins. Southern analysis of genomic DNA has shown that win1 and win2 are members of a small multi-gene family which is estimated to have a minimum of five members per haploid genome of Maris Piper and appears to be conserved within the Solanaceae. We have shown by Northern analysis and S1 mapping that the two genes exhibit differential organ-specific expression after the wounding of a potato plant.


Quinone formation from carcinogenic benzo[a]pyrene mediated by lipid peroxidation in phosphatidylcholine liposomes.
The behavior of benzo[a]pyrene (B[a]P) during peroxidation of phosphatidylcholine (PC) liposomes initiated by an azo compound was investigated to examine the mechanism of quinone formation from carcinogenic B[a]P mediated by nonenzymatic lipid peroxidation occurring in vivo. B[a]P had a retarding effect on the peroxidation of polyunsaturated fatty acid moiety of PC. The major oxidation products which accumulated in the peroxidized liposomes were B[a]P 1,6-, 3,6-, and 6,12-quinone. Antioxidants acting as scavengers of chain-propagating lipid peroxy radicals effectively prevented not only lipid peroxidation but also B[a]P oxidation in the liposomal suspension. PC hydroperoxides, the primary products of PC oxidation, did not react with B[a]P in the absence of the azo compound, indicating that lipid peroxy radicals, not lipid hydroperoxides, are responsible for the formation of these quinones. The experiments using 18O2 gas and 18O-labeled methyl linoleate hydroperoxides demonstrated that B[a]P quinones are formed by incorporating molecular oxygen and their origin is partly due to the lipid peroxy radical. The mechanism proposed for the formation of B[a]P quinones mediated by peroxidation of membrane lipids involves a direct attack of the lipid peroxy radical on B[a]P and subsequent autocatalytic oxidation. Weak carcinogenic and noncarcinogenic pentacyclic aromatic hydrocarbons showed little reactivity to the lipid peroxy radical in the liposomes. Thus, the facility of the peroxidative attack on B[a]P may be related to the powerful carcinogenic activity of this substance.


Urinary incontinence and hysterectomy in a large prospective cohort study in American women.
We investigated the effects of hysterectomy with and without concomitant urinary incontinence repair on incontinence severity in one of the largest prospective studies of hysterectomy outcomes performed in the United States. We administered the Urinary Symptoms Scale for Women to 1,299 women before hysterectomy done for benign conditions, and 6, 12, 18 and 24 months postoperatively to measure the presence, severity and type of urinary incontinence. Before hysterectomy 29.5% of participants had severe urinary incontinence, which decreased to 10% 12 and 24 months after hysterectomy. Most women with severe incontinence before hysterectomy showed improved incontinence 1 year after surgery (89.1%) and were still improved after 2 years (86.5%). The majority of women with moderate incontinence before hysterectomy showed improved incontinence 1 year after surgery (62.4%) and were still improved after 2 years (61.2%). However, some women with mild or no incontinence before hysterectomy had new onset incontinence or the condition had worsened 1 year after surgery (16.7%) and was still worse after 2 years (14.4%). A concomitant urinary incontinence repair procedure significantly increased the probability that a woman with severe incontinence would have improved incontinence 1 year after hysterectomy and significantly decreased the probability that a woman with mild or no incontinence would have worse incontinence after 2 years. The majority of women who undergo hysterectomy for nonmalignant indications experienced improved urinary incontinence during the first 2 years after surgery.


Long coats, short coats and no coats: chaplaincy presents to psychiatry at the University of Texas MD Anderson Cancer Center, a report.
When Chaplaincy and Psychiatry examine their own methodologies, do they work to reduce age-old barriers, thereby involving each other to promote holistic patient care? Chaplaincy trains in self-awareness and pastoral care specializing in religion, spirituality, grief and loss; while Psychiatry trains in medicine, neurology, and the behavioral neurosciences. Relationships across disciplines with common interests are vital. Ongoing dialogue between these professions will enhance the shared goals of coping and healing in the communities they serve.


Molecular cloning and expression of a mammalian homologue of a translationally controlled tumor protein (TCTP) gene from Penaeus monodon shrimp.
White spot syndrome, caused by white spot syndrome virus (WSSV), is a deadly disease of shrimps, causing a catastrophic loss in shrimp industries worldwide. In order to investigate molecular response of shrimp haemocyte to WSSV infection, we performed subtraction hybridization of mRNAs from healthy and WSSV-infected haemocyte. One of the genes that were severely down-regulated in moribund WSSV-infected-haemocyte was translationally controlled tumor protein (TCTP) (or fortilin). Strikingly, while there was a slight difference in the amount of TCTP message between normal and early WSSV-infected shrimps, shrimps that exhibited severe symptoms uniformly had very little TCTP in their haemocyte. Taken together with the fact that TCTP functions as an anti-apoptotic protein in mammals, our data suggest that TCTP in shrimp protects WSSV-infected shrimps from death.


Inhibition of new blood vessel formation in mice by systemic administration of human rib cartilage extract.
1 M HCl-guanidine extract of human funnel chest rib cartilage administered i.v. to mice decreased specifically vasoproliferation induced by intradermal injection of allogeneic murine lymphocytes.


Working in the dark: Interaction with a sub clinical COVID-19 subject and lessons learned.
Subclinical COVID-19 subjects pose a significant challenge. We present a very close clinical interaction with a subclinical COVID-19 subject that met the "standard screening criteria" and is unique in several ways. Learning from our experience, we suggest close attention should be paid to any unexpected findings such as groundglass opacity on CT as it could help early identification of subclinical COVID-19 infection.


Chondrocutaneous Bilateral Advancement Flap with Postoperative Radiation Therapy for a Helical Rim Keloid.
Keloids can be recalcitrant, and a well-planned treatment strategy is essential. Multiple ear piercings have recently become popular, particularly among younger age groups. Management of keloids that develop after piercing of the ear cartilage may be particularly problematic. Helical rim keloids are difficult to excise because of the complex, three-dimensional, cartilaginous structure of the helix and its thin and tightly adherent covering layer of skin. The chondrocutaneous advancement flap introduced by Antia and Buch may be a useful reconstructive option for a helical rim keloid after marginal loss of a segment of the helix as a result of trauma, a burn, or excision of a malignant tumor. However, this technique is limited to wounds that involve only the helix. In this technical note, we describe the use of a chondrocutaneous bilateral advancement flap with postoperative radiation therapy to treat a more invasive and relatively large keloid on the scapha. This technique is straightforward and safe in terms of preserving the blood supply. The addition of adjuvant radiation therapy can help to decrease the risk of recurrence and preserve the morphological structure of the ear and patient satisfaction.Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Masked transposition effects for simple versus complex nonalphanumeric objects.
When two letters/digits/symbols are switched in a string (e.g., jugde-judge; 1492-1942; *?$&-*$?&), the resulting strings are perceptually similar to each other and produce a sizable masked transposition priming effect with the masked priming same-different matching task. However, a parallel effect does not occur for strings of pseudoletters (e.g., [letters in the text]); García-Orza, Perea, & Muñoz, Quarterly Journal of Experimental Psychology, 63, 1603-1618, 2010). In the present study, we examined whether masked transposition priming is specific to alphanumeric stimuli or whether it also occurs with strings composed of other "objects"-namely, line drawings of common objects (Experiment 1) and geometrical shapes (Experiment 2). Results showed a significant masked transposition priming effect for geometrical shapes (e.g., [geometrical shapes in the text]), but not for line drawings of common objects (e.g., [symbols in the text]). These findings suggest that the mechanism involved in the coding of position in masked priming works only with perceptually simple, familiar "objects" (i.e., letters, numbers, symbols, or geometrical shapes), once their identities have been well ascertained.


Ameliorative effect of flaxseed oil against thiacloprid-induced toxicity in rats: hematological, biochemical, and histopathological study.
The present study was carried out to evaluate the hematological, biochemical, and histopathological changes due to thiacloprid toxicity, and the potential protective role of flaxseed oil in male Wistar albino rats. Subacute thiacloprid intoxication induced a significant increase in RBCs, Hb, PCV, and WBCs count, and bone marrow micronucleus (MN) formation. Moreover, there was a significant increase in serum biochemical parameters related to hepatic injury: alanine aminotransferase (ALT) and alkaline phosphatase (ALP). Serum total protein and albumin levels were significantly reduced. Thiacloprid increases tumor necrosis factor-alpha (TNF-α) and interleukine-2(IL-2). There was a significant decrease in glutathione-S-transferase, while the lipid peroxidation (MDA) and cytochrome P450 activity were significantly increased. Flaxseed oil coadministration partially retrieved the changes in all studied parameters. Thiacloprid induced histopathological liver damage, which was minimized as a result of flaxseed oil treatment. In general, it was concluded that, flaxseed oil able to protect against thiacloprid-induced hepatoxicity.


Characteristics of Iberian red deer (Cervus elaphus hispanicus) spermatozoa cryopreserved after storage at 5 degrees C in the epididymis for several days.
The aim of this study was to assess the influence of prolonged cold storage of Iberian red deer epididymides on post-thaw sperm characteristics. Thirty-seven pairs of testes, with attached epididymides, were collected during November and December. Spermatozoa from one of each of the pairs were immediately recovered, evaluated and frozen (control group). The remaining epididymides were cooled to 5 degrees C and stored for 12, 24, 48, 72 and 96 h (experimental groups), after which spermatozoa were collected and frozen as in the control group. After thawing, sperm motility, membrane and acrosome integrities, mitochondrial function and DNA damage were evaluated. The motility of spermatozoa stored in the epididymis for up to 96 h did not decrease significantly (P>0.05) but, after cryopreservation, a decline in sperm motility was seen in spermatozoa stored for 48 h, or later. A slower decrease in sperm membrane and acrosome integrities after cryopreservation were seen as storage time progressed. Some differences were seen when different methods were used to assess the same sperm parameter although changes followed similar patterns. This was the case for acrosome integrity (phase contrast microscopy versus fluorescent lectin) or membrane integrity (hypo-osmotic swelling test or nigrosin-eosin stain versus propidium iodide). We conclude that frozen-thawed spermatozoa of Iberian red deer recovered from epididymides stored at 5 degrees C have a good sperm quality (including motility) during less than 48 h of storage for most of the sperm parameters assessed.


Software-guided (PREVEDEL) cognitive stimulation to prevent delirium in hospitalised older adults: study protocol.
Delirium is a clinical condition characterised by acute and fluctuating deterioration of the cognitive state, generally secondary to an acute pathology. Delirium is associated with negative outcomes in older adults, such as longer hospitalisations, higher mortality, and short and medium-term institutionalisation. Randomised clinical trials have shown that delirium is preventable through non-pharmacological prevention measures, decreasing its incidence by 30-50%. These interventions include promoting physical activity, facilitating the use of glasses and hearing aids, cognitive stimulation, and providing frequent reorientation of time and space, among others. These measures are currently seldom applied in hospitals in Chile and around the world for reasons including the heavy workload of clinical staff, the lack of trained personnel, and in general the absence of a systematic implementation processes. We developed a software called PREVEDEL, which includes non-pharmacological strategies such as cognitive stimulation, early mobilisation, orientation, and pain assessment. We propose a randomised clinical trial to evaluate whether cognitive stimulation guided by PREVEDEL software prevents delirium status (full/subsyndromal delirium) in hospitalised older adults. A randomised controlled trial, with parallel, multicentre groups. We will recruite patients 65 years or older who have been hospitalised for less than 48 h in the general ward or the intermediate care units of four hospitals in Santiago, Chile. The participants in the intervention group will use a tablet with cognitive stimulation software for delirium prevention for five continuous days versus the control group who will use the tablet without the software. We will evaluate the incidence, duration, density of delirium, subsyndromal delirium with the Confusion Assessment Method, cognitive with the Montreal Cognitive Assessment, and functional status with the Functional Independence Measure at discharge. Moreover, we will evaluate the adherence to prevention measures, as well as demographic variables of interest. The use of cognitive PREVEDEL software could increase and improve the implementation of non-pharmacological prevention measures for delirium in hospitalised older adults, thus reducing its incidence and contributing to patients and health professionals. NCT05108207 ClinicalTrials.gov. Registered 4 November 2021.


Transcription factor STE12alpha has distinct roles in morphogenesis, virulence, and ecological fitness of the primary pathogenic yeast Cryptococcus gattii.
Cryptococcus gattii is a primary pathogenic yeast, increasingly important in public health, but factors responsible for its host predilection and geographical distribution remain largely unknown. We have characterized C. gattii STE12alpha to probe its role in biology and pathogenesis because this transcription factor has been linked to virulence in many human and plant pathogenic fungi. A full-length STE12alpha gene was cloned by colony hybridization and sequenced using primer walk and 3' rapid amplification of cDNA ends strategies, and a ste12alpha delta gene knockout mutant was created by URA5 insertion at the homologous site. A semiquantitative analysis revealed delayed and poor mating in ste12alpha delta mutant; this defect was not reversed by exogenous cyclic AMP. C. gattii parent and mutant strains showed robust haploid fruiting. Among putative virulence factors tested, the laccase transcript and enzymatic activity were down regulated in the ste12alpha delta mutant, with diminished production of melanin. However, capsule, superoxide dismutase, phospholipase, and urease were unaffected. Similarly, Ste12 deficiency did not cause any auxotrophy, assimilation defects, or sensitivity to a large panel of chemicals and antifungals. The ste12alpha delta mutant was markedly attenuated in virulence in both BALB/c and A/Jcr mice models of meningoencephalitis, and it also exhibited significant in vivo growth reduction and was highly susceptible to in vitro killing by human neutrophils (polymorphonuclear leukocytes). In tests designed to simulate the C. gattii natural habitat, the ste12alpha delta mutant was poorly pigmented on wood agar prepared from two tree species and showed poor survival and multiplication in wood blocks. Thus, STE12alpha plays distinct roles in C. gattii morphogenesis, virulence, and ecological fitness.


Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a neonatal unit: control of an outbreak using a new ADSRRS technique.
Extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae (EPKP) strains are frequently implicated in outbreaks in neonatal units. From April 2002 to January 2003, 149 neonates were colonized/infected with EPKP in the Neonatal Clinic of the Teaching Hospital at the Medical University of Gdańsk, Poland. A novel assay based on suppression of PCR, ADSRRS-fingerprinting, was successfully evaluated for typing EPKP isolates. The results showed that the genotypes of all outbreak-related strains were identical, which suggested that the outbreak originated from a single clone. This conclusion was confirmed by using different methods--RAPD and PFGE. The outbreak was stopped by adopting improved hygiene and instituting outbreak control measures.


Black women's experiences of gendered racial sexual objectification, body image, and depressive symptoms.
Black women navigate unique sexual objectification experiences and concerns about their bodies as a consequence of the race- and gender-based marginalization that they face. However, less is known about the influence of gendered racial sexual objectification experiences on Black women's mental health (i.e., depressive symptoms) or the contributions of key body image indicators (i.e., body surveillance and current-ideal body image discrepancy) that reflect Black women's engagement in monitoring and managing their bodies. We surveyed 1595 Black women to test our hypotheses that experiences of gendered racial sexual objectification (i.e., frequency and stress appraisal) would be positively associated with depressive symptoms and that body surveillance and current-ideal body image discrepancy would moderate this association. Analyses showed that more frequent experiences of gendered racial sexual objectification and higher stress appraisal of these experiences were significantly associated with more depressive symptoms. Furthermore, body surveillance and current-ideal body image discrepancy moderated the relation between gendered racial sexual objectification and depressive symptoms. Findings highlight how Black women's objectification and increased engagement in body monitoring and management practices are associated with their experiences of depressive symptoms, and thus, may negatively influence their mental health.


Solitary recurrence of prostate cancer surrounded by seminal vesicle/vas deferens-like epithelium.
Clinical recurrence of prostate cancer after curative treatment with a limited number of metastases is often termed as oligorecurrence. We report a case of solitary recurrence of prostate cancer surrounded by epithelium of the seminal vesicle or vas deferens. A 54-year-old man diagnosed with localized prostate cancer underwent radiation therapy. Six years later, imaging studies detected a solitary recurrence. We performed metastasectomy, and histopathological examination revealed the metastatic lesion surrounded by the epithelium of the seminal vesicle or vas deferens. Surgical resection achieved a complete biochemical response. We presented with a case of prostate cancer metastasis surrounded by the epithelium of the seminal vesicle or vas deferens.


Median nerve compression secondary to bifid reversed palmaris longus.
A case of median nerve compression in the forearm caused by a bifid reversed palmaris longus muscle is reported. We are not aware of any previous description of this particular anomaly associated with compression of the median nerve.


Characteristics of surface evapotranspiration and its response to climate and land use and land cover in the Huai River Basin of eastern China.
The Huai River Basin (HRB) has experienced significant climate and land use and land cover changes (LUCC) which have impacted the water cycle in recent times. However, little is understood about the impact of climate change and LUCC affecting evapotranspiration (ET). Thus, we investigate how surface ET response to climate change and LUCC in the HRB for the period from 2001 to 2014. ET and land cover types products (i.e., MOD16 and MCD12Q1, respectively) from MODerate-resolution Imaging Spectroradiometer (MODIS) were employed in this research. Water balance method and D20 pan evaporation data (Epan) as well as eddy covariance (EC) measurements were used to validate the MOD16 product, and the Theil-Sen's slope estimator and Mann-Kendall (M-K) test were adopted to estimate the magnitude and significance of ET trends. Moreover, daily meteorological data of 137 weather stations from 2001 to 2014 were also employed to explore the correlation mechanism between ET and meteorological factors. The results showed that the accuracy of MOD16 product data were convincible and could be used to estimate ET in the HRB. The higher values of ET are mainly distributed in the south and lower values in the north. ET decreased significantly in all seasons except in spring, especially in winter. The results also depicted that the land-use type in the HRB is mainly croplands, followed by forests and grasslands. Croplands area showed a decreasing trend at a rate of -176.2 km2·a-1, grasslands area presents a w-type fluctuation decreasing trend with a rate of -35.8 km2·a-1, urban/built-up area increased at a rate of 138.3 km2·a-1, water bodies area decreased gradually at a rate of -1.38 km2·a-1, wetlands area increased significantly at a rate of 43.6 km2·a-1, and barren area decreased gradually at a rate of -9.5 km2·a-1. The average annual ET is closely related to land-use types and shows a significant downward trend in general. The corresponding ET magnitude is exhibited as follows: forests>grasslands>croplands>wetlands>water bodies>urban/build-up lands>barren. Results of the study also suggest water conditions (precipitation (Pre) and relative humidity (RH) decrease) are major controlling factors in the decline of ET. Overall, the LUCC has a smaller influence on ET than climate change in the HRB. This research will provide a better insight into climate change and LUCC impacts on water resources.


A guide for endometrial cancer cell lines functional assays using the measurements of electronic impedance.
Endometrial cancer cell lines are critical tools to investigate the molecular mechanism of tumorigenesis using the end point cell-based assay such as proliferation, cytotoxicity, apoptosis, anoikis or migration and invasion. The proper assay optimization and performance is essential for physiologically relevant results interpretation. In this study we use label-free real-time cell analysis platform (xCELLigence) to optimize growing conditions for proliferation and migration experiments of two types of endometrial cancer cell lines HEC-1-B, HEC-1-A, KLE, and Ishikawa. Profiling of cell lines by cell index measurement in proliferation and migration experiments was performed. Our experimental approach allowed us to monitor particular stage of the cell growth, to see the relation between seeding density and dynamic cell growth as well as to choose the optimal serum concentration as chemoattractant in migration experiment. The highest rate of proliferation was shown for Ishikawa cells. The rapid pace of cellular migration was observed in case of KLE and HEC-1-B cells as compared to weak migratory activity of Ishikawa cells. The cell index that reflects the cell status characterized real-time cytological profile of each analyzed cell line. These cell profiles were crucial for better planning the classical end-point assays used in further research.


Do novel psychoactive substances displace established club drugs, supplement them or act as drugs of initiation? The relationship between mephedrone, ecstasy and cocaine.
To assess whether novel psychoactive substances (NPS) displace established club drugs, supplement them or act as drugs of initiation via a study of the relationship between mephedrone, ecstasy pills, cocaine and MDMA powder amongst club-goers considered to be 'early adopters' of psychostimulant/club drug trends. In situ surveys were conducted with 308 customers in two south London gay dance clubs across 3 weekend nights in July 2010 to assess the prevalence and patterns of self-reported use of a range of illegal drugs and NPS. Mephedrone was added to existing drug repertoires amongst those surveyed and acted to supplement more established club drugs including ecstasy pills, cocaine and MDMA powder, rather than replacing or displacing those drugs. This survey suggests that NPS are likely to be added to drug repertoires, particularly amongst experienced users with consequent health risks for individuals and resource implications for services. This study points to a complex relationship between NPS and illegal drug availability, purity and regulatory control, one which is increasingly important to understand given the global emergence of NPS and the challenges they present to existing supply, demand and harm reduction strategies.


Increased Visceral Adipose Tissue Is an Independent Predictor for Future Development of Atherogenic Dyslipidemia.
Atherogenic dyslipidemia is frequently observed in persons with a greater amount of visceral adipose tissue (VAT). However, it is still uncertain whether VAT is independently associated with the future development of atherogenic dyslipidemia. The aim of this study was to determine whether baseline and changes in VAT and subcutaneous adipose tissue (SAT) are associated with future development of atherogenic dyslipidemia independent of baseline lipid levels and standard anthropometric indices. Community-based prospective cohort study with 5 years of follow-up. A total of 452 Japanese Americans (240 men, 212 women), aged 34-75 years were assessed at baseline and after 5 years of follow-up. Abdominal fat areas were measured by computed tomography. Atherogenic dyslipidemia was defined as one or more abnormalities in high-density lipoprotein (HDL) cholesterol, triglycerides, or non-HDL cholesterol levels. Baseline VAT and change in VAT over 5 years were independently associated with log-transformed HDL cholesterol, log-transformed triglyceride, and non-HDL cholesterol after 5 years (standardized β = -0.126, 0.277, and 0.066 for baseline VAT, respectively, and -0.095, 0.223, and 0.090 for change in VAT, respectively). However, baseline and change in SAT were not associated with any future atherogenic lipid level. In multivariate logistic regression analysis, incremental change in VAT (odds ratio [95% confidence interval], 1.73 [1.20-2.48]; P = .003), triglycerides (4.01 [1.72-9.33]; P = .001), HDL cholesterol (0.32 [0.18-0.58]; P < .001), and non-HDL cholesterol (7.58 [4.43-12.95]; P < .001) were significantly associated with the future development of atherogenic dyslipidemia independent of age, sex, diastolic blood pressure, homeostasis model assessment insulin resistance, body mass index (BMI), change in BMI, SAT, and baseline atherogenic lipid levels. Baseline and change in VAT were independent predictors for future development of atherogenic dyslipidemia. However, BMI, waist circumference, and SAT were not associated with future development of atherogenic dyslipidemia.


Studies on the development of omasum in West African dwarf goats (Capra hircus).
This work studied the sequential morphological changes of omasum in foetuses, neonates and adult West African Dwarf (WAD) goats by gross and light microscopic methods. The mean omasal volume was 1.37 +/- 0.36, 4.2 +/- 0.4, 8.7 +/- 6.9 and 60.1 +/- 8.6 ml for gestation day 87 and 146 foetuses, neonates and adults respectively. Grossly the mucosa exhibited longitudinally oriented primary, secondary, tertiary and quartnery laminae with smooth surface in foetuses and numerous papillae in the adults. Microscopically it was lined by stratified squamous epithelium which was divided into larger lighter luminal and smaller darker basal zones in gestation day 60 foetuses. The core of the laminae contained extension of the inner muscular tunic. The basal zone developed lateral evaginations (corial papillae) into the lighter zone of the laminae in gestation day 106 foetuses. By term these corial papillae approached the luminal surface. The papillae emerged above the luminal surface in 4 week old neonates. The papillae were fully developed in adult goats. The mean papillary height and width were -205.1 +/- 34.0, 67.0 +/- 9.6; 235.0 +/- 86.5, 185.0 +/- 42.3 and 570.3 +/- 60.0, 290.1 +/- 66.3 microm for foetuses, neonates and adult goats. The inner circular layer was thicker than the outer longitudinal layer of the muscular tunic. The central muscular layer of the laminae originated from the inner circular layer with attachment to the outer muscular layer in adults. This attachment could be adaptation for better anchorage and muscular contraction of the laminae in WAD goats.


Determination of albuterol concentrations in human plasma using solid-phase extraction and high-performance liquid chromatography with fluorescence detection.
A fast, simple, and accurate method for determining albuterol concentrations in human plasma has been developed and validated for use in routine clinical analyses. This method involves a solid-phase extraction procedure using silica cartridges and normal-phase high-performance liquid chromatography with fluorescence detection. Regression analysis showed the method was linear over the standard curve range 1-16 ng ml-1. The percent recovery for albuterol and the internal standard, bamethane, at 5 ng ml-1 was found to be greater than 90%. The newly developed method has been applied in the analysis of plasma samples from patients and healthy volunteers.


Determination of methacrylic acid in food simulants by pyrolytic butylation-gas chromatography.
An on-line pyrolytic butylation approach was proposed to determine methacrylic acid (MA) in food simulants by gas chromatography (GC) without an expensive pyrolyzer. MA in food simulants was converted into butyl methacrylate in the presence of tetrabutylammonium hydroxide (TBAH) without any pretreatment at 330°C in the injection-port, contributing to high GC signal response. The derivatizing conditions for the proposed method were optimized, namely the injection-port temperature, type and amount of the organic alkaline used for derivatization. A series of standard solutions of MA in the range of 1.0-50mg/kg were analyzed with correlation coefficient r≥0.9975. The limits of detection (LODs) were less than 0.15mg/kg for MA in four matrix simulants (distilled water, 3%w/v acetic acid, 10%v/v ethanol, and isooctane). Relative standard deviations (RSDs) for retention time, peak height and peak area were all less than 3.88%. The technique was successfully applied to the analysis of MA migrating from plastic cup samples, with recoveries of added MA in the range of 96.5-123.0%. Direct injection of the simulants into the GC system after migration tests, without any pretreatment step, makes the developed method of great value for rapid screening analysis of samples in bulks.


Nephron-sparing surgery for renal cell carcinoma of the allograft after renal transplantation: report of two cases.
The risk of renal transplanation patients developing de novo malignancy is increased 100-fold compared with the healthy nontransplantation population. Renal cell carcinoma (RCC) arising from native kidneys is diagnosed among up to 4.6% of the renal transplant recipients as a consequence of immunosuppression. These tumors tend to behave more aggressively.(1) Although tumors occurring in allografted kidneys can be treated by partial (to save functional graft) or total nephrectomy, there is a paucity of data the outcomes. From 1978 to 2012, we performed 804 kidney transplantations including two cases in which RCC arose from the allografted kidney, both of which were treated with nephron-sparing surgery. The first patient has been followed for 30 months with a well functioning graft without an RCC recurrence. The second patient has returned to dialysis after 6 months due to an insufficient remnant nephron mass. In conclusion, nephron-sparing surgery is a novel alternative to total nephrectomy for allograft RCC. The remaining kidney can preserve function and the patient may not need chronic dialysis.


Quantitation of uterine activity preceding preterm, term, and postterm labor.
To assess uterine activity before labor in patients delivering preterm, at term, and postterm, the maximum spontaneous contraction frequency per 10-minute window during the initial portion of antepartum fetal heart rate monitoring was analyzed. Patients with multiple gestation, third trimester bleeding, polyhydramnios, or premature rupture of membranes and those already diagnosed with preterm labor were eliminated from the study. Of the 2446 remaining patients (7247 antepartum fetal heart rate tests) who went into spontaneous labor, 237 did so before 37 completed weeks of gestation, 1077 entered labor at term (38 to 42 completed weeks), and 1132 did so after 42 weeks. There was a significant increase in maximum uterine activity per 10-minute window from 30 to 44 weeks of gestation (average 4.7% per week; r = 0.97, p less than 0.0001). When compared with patients delivering spontaneously at term, average maximum uterine activity per 10-minute window was greatest in the preterm labor group (p less than 0.05) and least in the postterm labor group (p less than 0.05). These differences were present for several weeks preceding the onset of spontaneous labor. All three groups showed a surge of uterine activity during the 3 days before the onset of spontaneous labor.


Salicylate, mefenamate, meclofenamate, and quinine on cochlear potentials.
The perilymphatic spaces of guinea pig cochleae were perfused with artificial perilymph, with and without drug, at a rate of 2.5 microliters/minute for 10 minutes. The compound action potential of the auditory nerve, cochlear microphonics, and the summating potential evoked by 10 kHz tone bursts of varying intensities were recorded from a wire inserted in the basal turn scala vestibuli. The endocochlear potential was recorded from the scala media. Sodium salicylate (1.25 to 10 mmol/L) reduced the magnitude of the compound action potential evoked by low-sound intensities without affecting the compound action potential evoked by high-sound intensities. Sodium salicylate also reduced cochlear microphonics and had no effect on summating potential. Cochlear perfusions of prostaglandin synthesis inhibitors, mefenamate (200 mumol/L), and meclofenamate (200 mumol/L), had no effect on the cochlear potentials. Quinine (10 to 100 mumol/L) reduced the compound action potential input-output function in a parallel fashion rather than selectively affecting the low-intensity compound action potential. Quinine (100 mumol/L) reduced cochlear microphonics and summating potential. Neither quinine (100 mumol/L) nor salicylate (5 mmol/L) affected endocochlear potential. These results suggest that salicylate-induced hearing loss is not caused by either antagonism of the hair cell transmitter or cyclooxygenase inhibition, nor is it caused by the same mechanism that causes quinine-induced hearing loss.


An experimental model for ear reconstruction with moulded perichondrial flaps: a preliminary report.
Attempts to create a pinna by moulding cartilage fragments have been reported previously by Peer. The regenerative capabilities of perichondrium are well known. Combining these concepts, we succeeded in creating a cartilage pinna by implanting perichondrial flaps between the leaves of a methylmethacrylate mould. Ten rabbits were divided into two groups, the first using vascularised perichondrial flaps and the other using free perichondrium into which an arterial and venous pedicle was implanted. Both of these preparations were placed in the moulds in a subcutaneous pocket. Six weeks later, these constructions were harvested and histological examination conducted. Active cartilage growth was noted in both groups. The construction was completed by coverage with a split thickness skin graft. Clinically, this new procedure may provide a means of pinna reconstruction with a thin flexible cartilage framework, with excellent relief.


Uterotonic properties of the methanol extract of Monechma ciliatum.
The oxytocic activity of the hot methanol extract (HME) of the leaves of Monechma ciliatum was compared with other uterine stimulants like ergometrine, oxytocin, 5-hydroxytryptamine (5-HT), acetylcholine (ACh) and prostaglandins (PGs) E2 and F2alpha (PGE2 and PGF2alpha) in the presence of some antagonists in an attempt to explain the mechanism of action of the extract. The effects of the reference drugs on uteri isolated from rats pretreated with HME for 2 weeks were also observed. Atropine blocked the effect of ACh and partially blocked those of HME while L-366-948 blocked only the effect of oxytocin. Indomethacin inhibited the effects of HME as well as all the other drugs, except the PGs and ACh. D-600 blocked the effect of all the drugs including HME. Methysergide antagonised only the effect of 5-HT and partially blocked ergometrine. Prolonged treatment altered the uterine musculature and the activity profile of the reference drugs. These results suggest that the HME may be acting by more than one mechanism to contract the uterus and explains the mechanism of the anti-implantation activity of the plant.


Asiatic acid inhibits cardiac hypertrophy by blocking interleukin-1β-activated nuclear factor-κB signaling in vitro and in vivo.
Activated interleukin (IL)-1β signaling pathway is closely associated with pathological cardiac hypertrophy. This study investigated whether asiatic acid (AA) could inhibit IL-1β-related hypertrophic signaling, and thus suppressing the development of cardiac hypertrophy. Transverse aortic constriction (TAC) induced cardiac hypertrophy in C57BL/6 mice and cultured neonatal cardiac myocytes stimulated with IL-1β were used to evaluate the role of AA in cardiac hypertrophy. The expression of atrial natriuretic peptide (ANP) was evaluated by quantitative polymerase chain reaction (qPCR) and the nuclear factor (NF)-κB binding activity was measured by electrophoretic mobility shift assays (EMSA). AA pretreatment significantly attenuated the IL-1β-induced hypertrophic response of cardiomyocytes as reflected by reduction in the cardiomyocyte surface area and the inhibition of ANP mRNA expression. The protective effect of AA on IL-1β-stimulated cardiomyocytes was associated with the reduction of NF-κB binding activity. In addition, AA prevented TAC-induced cardiac hypertrophy in vivo. It was found that AA markedly reduced the excessive expression of IL-1β and ANP, and inhibited the activation of NF-κB in the hypertrophic myocardium. Our data suggest that AA may be a novel therapeutic agent for cardiac hypertrophy. The inhibition of IL-1β-activated NF-κB signaling may be the mechanism through which AA prevents cardiac hypertrophy.


Verification of nuclear data for the Tsukuba plan, a newly developed treatment planning system for boron neutron capture therapy.
Various verifications were performed to apply JENDL-4.0 as nuclear data for a newly developed treatment planning system with a homogeneous or precise human-like phantom. The nitrogen dose calculated by JENDL-4.0 differed slightly from that calculated by ENDF/B-VII.0. However, the total weighted dose-based dose volume histogram in the boron neutron capture therapy (BNCT) treatment for brain tumors calculated by JENDL-4.0 was in good agreement with the results of the ENDF/B-VII.0 calculation. Therefore, calculation with JENDL-4.0 can be applied to the BNCT dose calculation.


Preliminary characterization of porcine bone marrow stromal cells: skeletogenic potential, colony-forming activity, and response to dexamethasone, transforming growth factor beta, and basic fibroblast growth factor.
Neonatal pig bone marrow stromal cells (PBMSC) were tested in vivo and in vitro to establish their use as a large-animal model for the study of skeletogenesis. When implanted in diffusion chambers in athymic mice for 6-8 weeks, both freshly isolated pig bone marrow and passage 2 PBMSC formed partially mineralized cartilage, bone-like material, and fibrous tissue. The cartilage showed metachromatic, perilacunar staining with toluidine blue and safronin O, alcian blue staining for chondroitin and keratan sulfate, and intense immunostaining for type II collagen. Osteocalcin was immunolocalized to the mineralized regions, consistent with the formation of bone. Alkaline phosphatase was primarily observed in cell layers at boundaries between tissue types. Unstimulated monolayer cultures of PBMSC produced type I but not type II collagen, responded to dexamethasone (10(-8) M) with a 1.7-fold increase in alkaline phosphatase activity, and were stimulated to divide by basic fibroblast growth factor (1.5-fold; EC50 1 ng/ml). Transforming growth factor beta (TGF-beta) blocked both dexamethasone-induced alkaline phosphatase expression (EC50, 1 ng/ml of TGF-beta) and the mitogenic effects of bFGF (EC50 0.06 ng/ml of TGF-beta). When incubated for 10-14 days in medium containing dexamethasone, beta-glycerophosphate and ascorbate PBMSC formed mineralized nodules. Calcification occurred in the middle of the aggregates and was associated with intensely alkaline phosphatase positive cells and a dense type I collagen-rich matrix. PBMSC also displayed colony-forming unit-fibroblastic activity, with approximately 1 in 80 of the plated cells formed colonies > 128 cells over 14-21 days. PBMSC therefore mimic the known activities of stromal cells from other species, including the human, suggesting that they are a valid model for skeletal research.


Nanoformulation of Apolipoprotein E3-Tagged Liposomal Nanoparticles for the co-Delivery of KRAS-siRNA and Gemcitabine for Pancreatic Cancer Treatment.
KRAS is the most frequently mutated gene in human cancers, and ~ 90% of pancreatic cancers exhibit KRAS mutations. Despite the well-known role of KRAS in malignancies, directly inhibiting KRAS is challenging. In this study, we successfully synthesized apolipoprotein E3-based liposomes for the co-delivery of gemcitabine (GEM) and a small interfering RNA targeting KRAS (KRAS-siRNA) to improve the efficacy of pancreatic cancer treatment. Apolipoprotein E3 self-assembly on the liposome surface led to a substantial increase in its internalization in PANC1 human pancreatic cancer cells. KRAS-siRNA led to downregulated KRAS protein expression and KRAS-dependent carcinogenic pathways, resulting in the inhibition of cell proliferation, cell cycle arrest, increased apoptosis, and suppression of tumor progression. The combination of KRAS-siRNA and GEM induced a synergistic improvement in cell apoptosis and significantly lower cell viability compared with single-agent therapy. The low IC50 value of A3-SGLP might be attributed to potentiation of the anticancer effect of GEM by siRNA-mediated silencing of KRAS mutations, thereby inducing synergistic effects on cancer cells. A3-SGLP led to a marked decrease in the overall tumor burden and did not show any signs of toxicity. Therefore, the combination of KRAS-siRNA and GEM holds great potential for the treatment of pancreatic cancer.


Natural Vs Managed Habitat: Effect Over the Seed-Predator Pachymerus nucleorum and Its Natural Enemies.
The licuri palm, Syagrus coronata (Martius) Beccari (Arecaceae), is widely distributed throughout the Brazilian Caatinga and has high cultural, socioeconomical, and ecological importance. The palm tree logging is prohibited by the Brazilian law, and thus isolated individuals are a common sight on managed pastures in the Brazilian semi-arid region. We aimed to compare the insect seed-predator Pachymerus nucleorum (Fabricius) (Bruchinae) abundance and its predation levels on S. coronata seeds between managed (pasture) and natural (Caatinga vegetation) habitats. We also monitored the parasitoid Heterospilus prosopodis (Viereck) (Braconidae) abundance and other P. nucleorum potential natural enemies (generalist predators and microhymenopterans). We tested the hypothesis that more complex and heterogenous habitats (i.e., with higher plant diversity) support higher abundance of potential P. nucleorum natural enemies. For such, we collected 600 fruits from each habitat and evaluated the seed predation level by P. nucleorum, as well as the P. nucleorum parasitism by H. prosopodis. The P. nucleorum abundance and its potential natural enemies were estimated using 122 sticky traps placed on the S. coronata individuals' crown. Neither the P. nucleorum and generalist predators abundance differed between habitats, whereas the H. prosopodis and microhymenopterans abundance was higher in the natural habitat. Consequently, P. nucleorum parasitism levels by H. prosopodis were also higher in the natural habitat. Our study indicated that habitat with higher plant diversity supported more natural enemies, thus confirming that increased habitat homogenization leads to decreased parasitism levels by the less parasitoids number in managed habitats. Our results may subsidize conservationist management practices in the managed habitats aiming to improve fruit exploitation techniques sustainability and land-use practices, which would thereafter allow for the S. coronata population conservation in the Brazilian Caatinga.


Families at risk of poor parenting: a model for service delivery, assessment, and intervention.
The At Risk Parent Child Program is a multidisciplinary network agency designed for the secondary prevention of poor parenting and the extremes of child abuse and neglect. This model system of service delivery emphasizes (1) the coordination of existing community resources to access a target population of families at risk of parenting problems, (2) the provision of multiple special services in a neutral location (ambulatory pediatric clinic), and (3) the importance of intensive individual contact with a clinical professional who serves as primary therapist, social advocate and service coordinator for client families. Identification and assessment of families is best done during prenatal and perinatal periods. Both formal and informal procedures for screening for risk factors are described, and a simple set of at risk criteria for use by hospital nursing staff is provided. Preventive intervention strategies include special medical, psychological, social and developmental services, offered in an inpatient; outpatient, or in-home setting. Matching family needs to modality and setting of treatment is a major program concern. All direct services to at risk families are supplied by professionals employed within existing local agencies (hospital, public health department, state guidance center, and medical school pediatric clinic). Multiple agency involvement allows a broad-based screening capacity which allows thousands of families routine access to program services. The administrative center of the network stands as an independent, community-funded core which coordinates and monitors direct clinical services, and provides local political advocacy for families at risk of parenting problems.


Developmental arsenic exposure impairs cognition, directly targets DNMT3A, and reduces DNA methylation.
Developmental arsenic exposure has been associated with cognitive deficits in epidemiological studies, but the underlying mechanisms remain poorly understood. Here, we establish a mouse model of developmental arsenic exposure exhibiting deficits of recognition and spatial memory in the offspring. These deficits are associated with genome-wide DNA hypomethylation and abnormal expression of cognition-related genes in the hippocampus. Arsenic atoms directly bind to the cysteine-rich ADD domain of DNA methyltransferase 3A (DNMT3A), triggering ubiquitin- and proteasome-mediated degradation of DNMT3A in different cellular contexts. DNMT3A degradation leads to genome-wide DNA hypomethylation in mouse embryonic fibroblasts but not in non-embryonic cell lines. Treatment with metformin, a first-line antidiabetic agent reported to increase DNA methylation, ameliorates the behavioral deficits and normalizes the aberrant expression of cognition-related genes and DNA methylation in the hippocampus of arsenic-exposed offspring. Our study establishes a DNA hypomethylation effect of developmental arsenic exposure and proposes a potential treatment against cognitive deficits in the offspring of pregnant women in arsenic-contaminated areas.