Development and validation of a liquid chromatography-tandem mass spectrometric assay for pitavastatin and its lactone in human plasma and urine.
A rapid, selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with electrospray ionization (ESI) was developed and validated for the simultaneous determination of pitavastatin and its lactone in human plasma and urine. Following a liquid-liquid extraction, both the analytes and internal standard racemic i-prolact were separated on a BDS Hypersil C(8) column, using methanol-0.2% acetic acid in water (70: 30, v/v) as the mobile phase. The mass spectrometer was operated in multiple reaction monitoring (MRM) mode using the transition m/z 422.4-->m/z 290.3 for pitavastatin, m/z 404.3-->m/z 290.3 for pitavastatin lactone and m/z 406.3-->m/z 318.3 for the internal standard, respectively. Linear calibration curves of pitavastatin and its lactone were obtained in the concentration range of 1-200 ng/ml, with a lower limit of quantitation of 1 ng/ml. The intra- and inter-day precision values were less than 4.2%, and accuracies were between -8.1 and 3.5% for both analytes. The proposed method was utilized to support clinical pharmacokinetic studies of pitavastatin in healthy subjects following oral administration.


Sensitivity of ERG components from dark-adapted goldfish retinas treated with APB.
The electroretinogram (ERG) of the dark-adapted goldfish was examined before and after intravitreal injection of DL-2-amino-4-phosphonobutyric acid (APB). APB abolished the b-wave and decreased absolute sensitivity of the remaining waveform, which was composed of a vitreal-negative component followed by a vitreal-positive component. The sensitivity, time course and amplitude of these components differed from ERGs obtained from animals treated with sodium aspartate. Spectral sensitivity of both post-APB components closely resembled that of the normal dark-adapted b-wave. The results suggest that APB does not act selectively on any particular class of photoreceptors or photoreceptor pathways in the dark-adapted goldfish retina.


Construction of MnO2 Artificial Leaf with Atomic Thickness as Highly Stable Battery Anodes.
The leaf-like structure is a classic and robust structure and its unique vein support can reduce structural instability. However, biomimetic leaf structures on the atomic scale are rarely reported due to the difficulty in achieving a stable vein-like support in a mesophyll-like substrate. A breathable 2D MnO2 artificial leaf is first reported with atomic thickness by using a simple and mild one-step wet chemical method. This homogeneous ultrathin leaf-like structure comprises of vein-like crystalline skeleton as support and amorphous microporous mesophyll-like nanosheet as substrate. When used as an anode material for lithium ion batteries, it first solves the irreversible capacity loss and poor cycling issue of pure MnO2 , which delivers high capacity of 1210 mAh g-1 at 0.1 A g-1 and extremely stable cycle life over 2500 cycles at 1.0 A g-1 . It exhibits the most outstanding cycle life of pure MnO2 and even comparable to the most MnO2 -based composite electrode materials. This biomimetic design provides important guidelines for precise control of 2D artificial systems and gives a new idea for solving poor electrochemical stability of pure metal oxide electrode materials.


The triggers of the cGAS-STING pathway and the connection with inflammatory and autoimmune diseases.
Cyclic GMP-AMP synthase (cGAS) is a cytosolic nucleic acid sensor that can bind to dsDNA. It maintains an autoinhibited state in the absence of cytosolic dsDNA, while when activated, it in turn activates its adaptor protein STING, ultimately triggering a cascade that produces inflammatory cytokines and type I interferons (IFNs). With further research, additional types of nucleic acids have been found to be activators of the cGAS-STING pathway. The cGAS-STING pathway can provide protection or resistance against infections; however, improper or overactivation might cause severe inflammatory pathologies, including autoimmunity. This article systematically reviews the latest research progress on the axis, including categorical pathway triggers, the connection with autoimmune disease and drug therapy progress.


Extended embryo culture reduces the implantation rate on day 4 and day 5 when only a maximum of three embryos are cultured beyond the pronuclear stage.
To study the potential of embryo transfer after 3, 4 or 5 days of embryo culture under the German embryo protection law according to which only a maximum of three zygotes are allowed to be cultured for embryo transfer. In a prospective study, 273 patients with assisted reproductive treatment were randomly allocated for transfer on days 3, 4 or 5. Pregnancy and implantation rates were evaluated in regard to day of transfer and results were compared by Chi-square or ANOVA test. Out of 234 transfer cycles, 79 were performed on day 3, 76 on day 4 and 79 on day 5. Pregnancy and implantation rates were 41.8%/27.1% for transfer on day 3, 27.6%/14.1% for day 4 transfer and 16.5%/8.8% for transfer on day 5. These results were significantly different for pregnancy rates on day 3 versus day 5 (P < 0.001) and for implantation rates on day 3 versus day 4 (P < 0.005) and day 3 versus day 5 (P < 0.001). These findings suggest that extended embryo culture is not beneficial when the option for embryo selection at later stages of development is not available.


Factors influencing the age at natural menopause.
To examine sociodemographic, menstrual, reproductive, and other factors which may influence the age at natural menopause, the authors analyzed data from a large series of women participating in a nationwide breast cancer screening program conducted between 1973 and 1980. Standard life table techniques permitted assessment of factors suspected of varying the time to menstrual cessation among 983 premenopausal women, 1091 surgically menopausal women, and 1423 naturally menopausal women. The median age at natural menopause was 51.1 years. Multivariate analysis indicated that parity, irregularity of menstrual cycles before age 25 or first livebirth, and high socioeconomic status were significantly related to menopausal age. These data provide evidence for the hypothesis that certain environmental and hormonal factors which affect ovulation during reproductive years may ultimately postpone the menopause.


Pharmacokinetics of ciprofloxacin.
The fluorination of piperazinyl substituted quinolones has led to an interesting development of a series of new broad spectrum antibacterial agents that may be administered orally as well as parenterally and are well tolerated. Norfloxacin was an early compound, later followed by ciprofloxacin, enoxacin, ofloxacin and pefloxacin. In this overview the emphasis will be on the most extensively studied compound including comparisons, where data are available, with norfloxacin and ofloxacin. Enoxacin and pefloxacin will be omitted due to their pattern of side effects, which at present curtail their therapeutic use. More recent substances such as fleroxacin and defloxacin have not been sufficiently investigated to be considered in this context.


Fat Grafting for Facial Contouring (Temporal Region and Midface).
In this article the authors introduce a compartment-based fat graft for facial contouring, focusing on the anatomy of temporal region and midface, the 3L3M integrated fat transfer technique, and the facial compartment-based fat injection in the face. This article includes details of pre-evaluation methods, results of cases, and postoperation care. Readers will have a clear view of how to do facial contouring fat graft in the face after reading this article. There seldom are complications while using the 3L3M technique for fat grafting. The targeted fat grafting can lead to steady and efficient results.


Simultaneous and individual determination of seven biochemical species using a glassy carbon electrode modified with a nanocomposite of Pt nanoparticle and graphene by a one-step electrochemical process.
We propose a sensitive and selective electrochemical approach for simultaneous and individual determination of seven electroactive biochemical species using a modified electrode with a nanocomposite of Pt nanoparticles and reduced graphene oxides (PtNP/rGO). The PtNP/rGO nanocomposite was synthesized and deposited on a glassy carbon electrode (GCE) from Pt4+ precursors and GO by one-pot electrochemical synthesis, forming PtNP/rGO-GCE. As-prepared PtNP/rGO electrode was characterized by Raman spectroscopy, contact angle measurements, SEM-EDS analysis, and cyclic voltammetry. Compared with bare GCE, Pt electrode, PtNP-GCE, the modified electrode exhibited excellent catalytic activity and large surface area, and rGO-GCE, which enabled sensitive and selective detection of seven biochemical species such as ascorbic acid (AA), dopamine (DA), uric acid (UA), acetaminophen (AP), xanthine (XA), nitrite (NO2-), and hypoxanthine (HX) with good voltammetric resolution in differential pulse voltammetric (DPV) measurements. In addition, the electroanalytical performance of the PtNP/rGO-GCE sensor displayed satisfactory reproducibility and stability. Finally, the sensor could be applied for the detection of the seven biochemical species in real samples like human blood serum. Therefore, the PtNP/rGO-GCE can provide one promising platform for the simultaneous detection of redox-active biochemical species in environmental, food, and clinical samples.


Localization of the extracellular matrix protein SC1 coincides with synaptogenesis during rat postnatal development.
SC1 is an extracellular matrix protein that belongs to the SPARC family of matricellular molecules. This anti-adhesive protein localizes to synapses in the adult rat brain and has been postulated to modulate synapse shape. In this study, increased levels of SC1 were detected from postnatal days 10-20, with a peak at postnatal day 15, a period of intense synaptogenesis. During this time, increased colocalization of SC1 with the synaptic marker synaptophysin was observed in synapse-rich regions of the cerebellum and the cerebral cortex. These findings indicate that the pattern of SC1 localization coincided with synaptogenesis during rat postnatal development.


Quantitative measurement of humero-acromial, humero-coracoid, and coraco-clavicular intervals for the diagnosis of subacromial and subcoracoid impingement of shoulder joint.
The aim of this study was to determine the acromio-humeral, coraco-humeral, and coraco-clavicular distances with magnetic resonance imaging (MRI) in the diagnosis of shoulder impingement syndrome. All the acromio-humeral, coraco-humeral, and coraco-clavicular intervals of all patients were reviewed retrospectively. A total of 132 shoulders (76 right and 56 left shoulders) with impingement syndrome (40 males and 92 females, with 79 cases surgically confirmed and the remaining 53 cases clinically and radiologically suspected) and 40 control group patients were included. Statistical correlation of this research was assessed by Fisher's Exact t test and Pearson chi-square test-correlation coefficients. In the correlation of average acromio-humeral, coraco-humeral, and coraco-clavicular intervals (AHI, CHI, and CCI, respectively), there were significant statistical differences between patient and control groups; all the intervals were found to be higher in the control group. A moderate positive relation between average AHI and CHI, between AHI and CCI, and between CCI and CHI has been found. All intervals in the patient group increase or decrease dependently on each other. No proper cut-off values were determined using a receiver operating characteristic curve for all intervals between patient and control groups. Acromio-humeral, coraco-humeral, and coraco-clavicular intervals were extremely lower in the shoulder impingement syndrome and had significant importance in the diagnosis of subacromial and subcoracoid impingements.


Allogeneic vs. autologous mesenchymal stem/stromal cells in their medication practice.
Mesenchymal stem/stromal cell (MSC)-based therapeutics is already available for treatment of a range of diseases or medical conditions. Autologous or allogeneic MSCs obtained from self or donors have their own advantages and disadvantages in their medical practice. Therapeutic benefits of using autologous vs. allogeneic MSCs are inconclusive. Transplanted MSCs within the body interact with their physical microenvironment or niche, physiologically or pathologically, and such cells in a newly established tissue microenvironment may be impacted by the pathological harmful environmental factors to alter their unique biological behaviors. Meanwhile, a temporary microenvironment/niche may be also altered by the resident or niche-surrounding MSCs. Therefore, the functional plasticity and heterogeneity of MSCs caused by different donors and subpopulations of MSCs may result in potential uncertainty in their safe and efficacious medical practice. Acknowledging a connection between MSCs' biology and their existing microenvironment, donor-controlled clinical practice for the long-term therapeutic benefit is suggested to further consider minimizing MSCs potential harm for MSC-based individual therapies. In this review, we summarize the advantages and disadvantages of autologous vs. allogeneic MSCs in their therapeutic applications. Among other issues, we highlight the importance of better understanding of the various microenvironments that may affect the properties of niche-surrounding MSCs and discuss the clinical applications of MSCs within different contexts for treatment of different diseases including cardiomyopathy, lupus and lupus nephritis, diabetes and diabetic complications, bone and cartilage repair, cancer and tissue fibrosis.


Simple and efficient solid phase extraction based on molecularly imprinted resorcinol-formaldehyde resin nanofibers for determination of trace sulfonamides in animal-origin foods.
Novel molecularly imprinted resorcinol-formaldehyde resin nanofibers (MIRF NFs) was prepared using polydopamine as an intermediate. With high specific surface area and favorable usability, MIRF NFs presented high efficiency for the solid phase extraction (SPE) of sulfonamides (SAs) in complex animal foods. After optimizing the operating conditions, a new method for SAs quantification coupled with HPLC-MS/MS was developed. By simple water dilution of the solvent extracts, SPE could be carried out. In addition, the eluent could be analyzed directly without any further treatment. The newly developed method was simplified greatly with much fewer sample pretreatment procedures (4 steps). Moreover, much fewer amounts of sample (1.0 g), adsorbent (3.0 mg), organic solvent (1.5 mL) and preparation time (20 min for 24 samples) were needed. The obtained good linearity (R2 > 0.9957), low detection limits (0.01-0.14 μg kg-1), satisfactory recoveries (83.0 %-112.9 %) and precisions (RSDs < 12.6 %) further proved the feasibility of the method in practical application.


Anakinra combined with methylprednisolone in patients with severe COVID-19 pneumonia and hyperinflammation: An observational cohort study.
Immunomodulants have been proposed to mitigate severe acute respiratory syndrome coronavirus 2-induced cytokine storm, which drives acute respiratory distress syndrome in coronavirus disease 2019 (COVID-19). We sought to determine efficacy and safety of the association of IL-1 receptor antagonist anakinra plus methylprednisolone in severe COVID-19 pneumonia with hyperinflammation. A secondary analysis of prospective observational cohort studies was carried out at an Italian tertiary health care facility. COVID-19 patients consecutively hospitalized (February 25, 2020, to March 30, 2020) with hyperinflammation (ferritin ≥1000 ng/mL and/or C-reactive protein >10 mg/dL) and respiratory failure (oxygen therapy from 0.4 FiO2 Venturi mask to invasive mechanical ventilation) were evaluated to investigate the effect of high-dose anakinra plus methylprednisolone on survival. Patients were followed from study inclusion to day 28 or death. Crude and adjusted (sex, age, baseline PaO2:FiO2 ratio, Charlson index, baseline mechanical ventilation, hospitalization to inclusion lapse) risks were calculated (Cox proportional regression model). A total of 120 COVID-19 patients with hyperinflammation (median age, 62 years; 80.0% males; median PaO2:FiO2 ratio, 151; 32.5% on mechanical ventilation) were evaluated. Of these, 65 were treated with anakinra and methylprednisolone and 55 were untreated historical controls. At 28 days, mortality was 13.9% in treated patients and 35.6% in controls (Kaplan-Meier plots, P = .005). Unadjusted and adjusted risk of death was significantly lower for treated patients compared with controls (hazard ratio, 0.33, 95% CI, 0.15-0.74, P = .007, and HR, 0.18, 95% CI, 0.07-0.50, P = .001, respectively). No significant differences in bloodstream infections or laboratory alterations were registered. Treatment with anakinra plus methylprednisolone may be a valid therapeutic option in COVID-19 patients with hyperinflammation and respiratory failure, also on mechanical ventilation. Randomized controlled trials including the use of either agent alone are needed to confirm these results.


Unaltered graft survival and intragraft lymphocytes infiltration in the cardiac allograft of Cxcr3-/- mouse recipients.
Previous studies showed that absence of chemokine receptor Cxcr3 or its blockade prolong mouse cardiac allograft survival. We evaluated the effect of the CXCR3 receptor antagonist MRL-957 on cardiac allograft survival, and also examined the impact of anti-CXCR3 mAb in human CXCR3 knock-in mice. We found only a moderate increase in graft survival (10.5 and 16.6 days, p < 0.05) using either the antagonist or the antibody, respectively, compared to control (8.7 days). We re-evaluated cardiac allograft survival with two different lines of Cxcr3(-/-) mice. Interestingly, in our hands, neither of the independently derived Cxcr3(-/-) lines showed remarkable prolongation, with mean graft survival of 9.5 and 10.8 days, respectively. There was no difference in the number of infiltrating mononuclear cells, expansion of splenic T cells or IFN-gamma production of alloreactive T cells. Mechanistically, an increased other chemokine receptor fraction in the graft infiltrating CD8 T cells in Cxcr3(-/-) recipients compared to wild-type recipients suggested compensatory T-cell trafficking in the absence of Cxcr3. We conclude Cxcr3 may contribute to, but does not govern, leukocyte trafficking in this transplant model.


Increased Activity of a NK-Specific CAR-NK Framework Targeting CD3 and CD5 for T-Cell Leukemias.
NK effector cells expressing a CAR construct may be used to target T-lineage markers. In this work, we compared the activity of a NK-specific CAR-NK and a CAR-T framework when expressed on NK effector cells to target CD3 and CD5 in T-cell malignancies. Our results show that CD3-CAR-T is more active than CD5-CAR-T to eliminate malignant T cells in vitro, however, CD3-CAR-T were less efficient to eliminate tumor cells in vivo, while CD5-CAR-T had antitumor activity in a diffuse xenograft model. Lack of in vivo efficacy correlated with downregulation of CD3 levels in target T cells after coculture with CD3-CAR effector cells. The CAR-NK framework greatly improved the efficacy of CARs leading to increased degranulation, cytokine secretion and elimination of the tumor xenograft by CD5-CAR-NK effector cells. Finally, all CAR constructs were similarly effective to eliminate malignant T cells in vitro. Our results show that the NK-CAR framework improves the activity of CARs in NK cells and that CD5 would be a better target than CD3 for T-cell malignancies, as dynamic downregulation of target expression may affect in vivo efficacy.


Peptide T does not affect induction of pineal N-acetyltransferase by vasoactive intestinal peptide.
It has been suggested that the HIV virus binds to VIP recognition sites which can be blocked by the octapeptide, peptide T. Stimulation of VIP receptors on pinealocytes activates adenylate cyclase and increases the activity of the enzyme serotonin N-acetyltransferase (NAT). We examined whether peptide T or D-Ala peptide T amide affected this induction. We found no evidence for peptide T interference with NAT induction and conclude that if peptide T inhibits attachment of HIV virus to VIP receptors, it does so at regions other than that occupied by VIP in stimulating adenylate cyclase and NAT.


Fast sorption measurements of volatile organic compounds on building materials: Part 1 - Methodology developed for field applications.
A Proton Transfer Reaction-Mass Spectrometer (PTR-MS) has been coupled to the outlet of a Field and Laboratory Emission Cell (FLEC), to measure volatile organic compounds (VOC) concentration during a sorption experiments (Rizk et al., this issue) [1]. The limits of detection of the PTR-MS for three VOCs are presented for different time resolution (2, 10 and 20 s). The mass transfer coefficient was calculated in the FLEC cavity for the different flow rates. The concentration profile obtained from a sorption experiment performed on a gypsum board and a vinyl flooring are also presented in comparison with the profile obtained for a Pyrex glass used as a material that do not present any sorption behavior (no sink). Finally, the correlation between the concentration of VOCs adsorbed on the surface of the gypsum board at equilibrium (Cse ) and the concentration of VOCs Ce measured in the gas phase at equilibrium is presented for benzene, C8 aromatics and toluene.


Journal of Biocommunication Showcase: Recipient of AMI's 2019 Lifetime Achievement Award.
This JBC Showcase features the award winning medical illustration of Cynthia Turner. Cynthia was the recipient of the Association of Medical Illustrators' 2019 Lifetime Achievement Award. Her award was presented at the Awards Ceremony during the AMI's 2019 annual meeting held in Milwaukee, Wisconsin.


Mature cystic teratoma of the pancreas. Case report and review of the literature of a rare pancreatic cystic lesion.
Pancreatic cystic lesions are increasingly recognized and comprise different pathological entities. The management of these lesions is challenging, because of inadequate preoperative histological diagnosis. Among this family of lesions, mature cystic teratomas are an extremely rare finding. We present the case of a 61-year-old man with a mature cystic teratoma of the pancreas' uncinate process, incidentally discovered at diagnostic imaging. This case highlights the difficulty to obtain a preoperative diagnosis of this pathological entity and the need of increased awareness about mature cystic teratoma when examining a pancreatic cystic lesion.


Negative predictive value of dobutamine stress echocardiography for perioperative risk stratification in patients with cardiac risk factors and reduced exercise capacity undergoing non-cardiac surgery.
Guidelines recommend functional testing for myocardial ischaemia in the perioperative setting in patients with greater than one recognised cardiac risk factor and self-reported reduced exercise capacity. To determine the clinical utility of dobutamine stress echocardiography (DSE) for perioperative risk stratification in patients undergoing major non-cardiac surgery. Data on 79 consecutive patients undergoing DSE for perioperative risk stratification at a single centre were retrospectively reviewed to determine rates of major adverse cardiac events (MACE) during the index hospitalisation and 30 days post-discharge. Echocardiography and outcome data were obtained through a folder audit and echolab database. Out of the 79 DSE performed for perioperative risk stratification, 11 (14%) were positive (DSE +ve) and 68 (86%) were negative (DSE -ve). Management in the DSE +ve group included medical optimisation without invasive intervention (n = 7(64%)), diagnostic coronary angiography (n = 3(27%)) and coronary artery bypass graft (n = 1(9%)). None of the patients underwent percutaneous coronary intervention preoperatively. Perioperative MACE in the DSE +ve group was 36% compared to 4% in the DSE-ve group (P = 0.006). DSE +ve was a powerful predictor of perioperative inpatient MACE (OR 12.4, 95% CI 2.3-67, P = 0.003). The positive predictive value of DSE +ve status was 36%, whereas the negative predictive value of DSE-ve status for perioperative MACE was 96%. DSE for perioperative risk stratification had a high clinical utility in patients undergoing major non-cardiac surgery. In particular, a normal DSE had a high negative predictive value for perioperative MACE.


Effects of Nonimmersive Virtual Reality Intervention on Children With Spastic Cerebral Palsy: A Meta-analysis and Systematic Review.
This review aimed to assess the effectiveness of nonimmersive virtual reality intervention compared with traditional rehabilitation in improving the functions of the upper and lower limbs, balance, and social participation among children with spastic cerebral palsy. We used librarian-designed searches of 10 databases to identify research articles on randomized controlled trials that assessed the effectiveness of nonimmersive virtual reality in intervening spastic cerebral palsy patients up to April 15, 2023. Independent evaluation was conducted by two trained investigators using the evaluation criteria of randomized controlled trial quality indicated in the Cochrane Manual of Assessment "risk-of-bias tool." The Physical Therapy Evidence Database scale was used to evaluate the method and quality of the literature. Twenty-one research articles involving 779 patients with spastic cerebral palsy were included. Significant differences between the nonimmersive virtual reality rehabilitation and traditional rehabilitation groups were observed in all indicators, except for the Jebsen-Taylor Hand Function Test. Nonimmersive virtual reality intervention is effective in improving the function of the lower extremity, balance, and social participation in children with spastic cerebral palsy, but its effect on upper limb function is still controversial.


The Initiator Methionine tRNA Drives Secretion of Type II Collagen from Stromal Fibroblasts to Promote Tumor Growth and Angiogenesis.
Expression of the initiator methionine tRNA (tRNAi(Met)) is deregulated in cancer. Despite this fact, it is not currently known how tRNAi(Met) expression levels influence tumor progression. We have found that tRNAi(Met) expression is increased in carcinoma-associated fibroblasts, implicating deregulated expression of tRNAi(Met) in the tumor stroma as a possible contributor to tumor progression. To investigate how elevated stromal tRNAi(Met) contributes to tumor progression, we generated a mouse expressing additional copies of the tRNAi(Met) gene (2+tRNAi(Met) mouse). Growth and vascularization of subcutaneous tumor allografts was enhanced in 2+tRNAi(Met) mice compared with wild-type littermate controls. Extracellular matrix (ECM) deposited by fibroblasts from 2+tRNAi(Met) mice supported enhanced endothelial cell and fibroblast migration. SILAC mass spectrometry indicated that elevated expression of tRNAi(Met) significantly increased synthesis and secretion of certain types of collagen, in particular type II collagen. Suppression of type II collagen opposed the ability of tRNAi(Met)-overexpressing fibroblasts to deposit pro-migratory ECM. We used the prolyl hydroxylase inhibitor ethyl-3,4-dihydroxybenzoate (DHB) to determine whether collagen synthesis contributes to the tRNAi(Met)-driven pro-tumorigenic stroma in vivo. DHB had no effect on the growth of syngeneic allografts in wild-type mice but opposed the ability of 2+tRNAi(Met) mice to support increased angiogenesis and tumor growth. Finally, collagen II expression predicts poor prognosis in high-grade serous ovarian carcinoma. Taken together, these data indicate that increased tRNAi(Met) levels contribute to tumor progression by enhancing the ability of stromal fibroblasts to synthesize and secrete a type II collagen-rich ECM that supports endothelial cell migration and angiogenesis.


Repolarization Patterns in Congenital Heart Disease.
The aim of this study was to study the repolarization patterns in pediatric patients with cyanotic and acyanotic congenital heart diseases as prolonged QT indicates a myocardium at risk of ventricular arrhythmia. A cross-sectional case-control study included 50 patients with acyanotic congenital heart diseases and 50 patients with cyanotic congenital heart diseases who presented to Catheterization Unit of Cairo University Pediatric Hospital between March 2013 and June 2014. We included 50 healthy children as a control. For all the patients' measurement of oxygen saturation, echocardiography and 12-lead electrocardiogram (ECG) were done and the corrected QT (QTc) was measured. The mean QTc was significantly higher in acyanotic congenital heart diseases with volume overload than in control: 0.426 versus 0.4 s (p = 0.009). Increased left ventricular end-diastolic dimension was significantly associated with QTc prolongation (p = 0.01). Early repolarization was higher in congenital heart diseases (18 % in acyanotic patient, 48 % in cyanotic patients) than in control 6 %. Decreased oxygen saturation was significantly associated with early repolarization (p = 0.01). Prolonged QTc was higher in acyanotic congenital heart diseases with volume overload and increased left ventricular end diastolic dimension was a significant association. Decreased oxygen saturation was a significant association.


Baseline measures of primary health care team functioning and overall primary health care performance at Du Noon Community Health Centre.
Research consistently demonstrates the importance of effective team work for improving the quality of health care. We conducted a baseline measure of primary health care (PHC) team effectiveness and overall PHC performance at a primary care facility. To improve PHC team effectiveness and ultimately the quality and user experience of primary care at a community health centre (CHC). Du Noon CHC in the southern and western substructure of the Cape Town Metro district services (MDHS). A cross-sectional study using a combination of the Nominal Group Technique (NGT) consensus method and the South African Primary Care Assessment Tool (ZA PCAT) to assess PHC team effectiveness and PHC organisation and performance. The ZA PCAT was administered to 110 CHC users (patients) and 12 providers (doctors and clinical nurse practitioners). Data from 20 PHC team members showed they perceived their team as well functioning (70% agreement on a 7-item PHC team assessment tool incorporated into the ZA PCAT). The NGT method achieved participant (20) consensus on communication and leadership as the main challenges to effective team functioning and on ideas to overcome the challenges. The ZA PCAT user data showed 18.2% of users rated first contact access as acceptable to good; 47.3% of users rated ongoing care as acceptable to good. Provider data showed that 33% of providers rated first contact access as acceptable to good; 25% of providers rated ongoing care as acceptable to good. First contact access received the lowest acceptable to good score (18.2%) and comprehensiveness (services available) the highest score (88.2%) from users. For the providers, the lowest acceptable to good score was for ongoing care (25%) and the highest acceptable to good score was for primary health care team availability (100%). The ZA PCAT total primary scores were good (above 60%) for both users and providers but moderately higher for the providers. Knowledge of how teams perceive their effectiveness can motivate them to generate ideas for improving performance. There were discrepancies between providers' assessment of team functioning using the ZA PCAT measure and the NGT method results. The ZA PCAT also showed differences between providers' and users' perceptions of PHC performance - consistent with the findings of the multi-CHC Western Cape ZA PCAT study. These findings should encourage and support CHC and district level staff in their efforts to improve the quality and user experience of primary care, as well as PHC team performance.


Purkinje cell responses to pontine stimulation CS during rabbit eyeblink conditioning.
Previous studies have shown that stimuli typically used as CSs in eyeblink conditioning converge with US information in the cerebellum. Extracellular recordings of Purkinje cells have shown learning-related as well as stimulus-related discharge patterns. Stimulation of a portion of the auditory CS pathway, the pontine nucleus, also serves as a highly effective CS. Using a short-latency pontine stimulation CS and air puff US, single Purkinje cell responses were recorded and compared to those elicited with an auditory stimulus in previous work. Purkinje cell recordings in trained and untrained rabbits revealed patterns of responses very similar to those seen in rabbits trained to a tone CS or those given unpaired-tone/air-puff training. Similarities included the proportion of stimulus-related and behavior-related cell responses. However, fewer inhibitory responses were seen than in earlier studies and these differences are considered in light of the differences between an extracellular stimulation CS and a peripherally administered auditory CS.


Endovascular Management of Acute Pulmonary Embolism Using the Ultrasound-Enhanced EkoSonic System.
Acute, symptomatic pulmonary embolism (PE) in the massive and submassive categories continues to be a healthcare concern with significant risk for increased morbidity and mortality. Despite increased awareness and venous thromboembolism prophylaxis, endovascular treatment is still an important option for many of these patients. There are a variety of techniques and devices used for treating PE, but none have been evaluated as extensively as the EkoSonic endovascular system that is also currently the only FDA-approved device for the treatment of pulmonary embolism. This article describes the use of the EkoSonic device for this patient population.


Major influencing factors on routine implementation of shared decision-making in cancer care: qualitative process evaluation of a stepped-wedge cluster randomized trial.
Shared decision-making (SDM) is highly relevant in oncology but rarely implemented in routine care. In a stepped-wedge cluster randomized implementation trial, the outcome evaluation of a theoretically and empirically based multi-component SDM implementation program did not show a statistically significant effect on patient-reported SDM uptake. Within this SDM implementation trial, a thorough a priori planned process evaluation was conducted. Thus, the aim of this study was to investigate factors influencing SDM implementation in the context of a multi-component SDM implementation program. We conducted qualitative process evaluation of a stepped-wedge SDM implementation trial. Qualitative data included interviews with nurses and physicians of participating departments, field notes by the study team, and meeting minutes. Data were analyzed via deductive and inductive qualitative content analysis on basis of the Consolidated Framework for Implementation Research (CFIR). Transcripts of 107 interviews with 126 nurses and physicians, 304 pages of field note documentation, and 125 pages of meeting minutes were analyzed. Major factors influencing SDM implementation were found for all domains of the CFIR: a) four regarding characteristics of the individuals involved (e.g., perceived personal relevance, individual motivation to change), b) eleven regarding the inner setting (e.g., leadership engagement, networks and communication, available resources, compatibility with clinical practice), c) two regarding the outer setting (e.g., culture of health care delivery), d) eight regarding characteristics of the intervention (e.g., relative advantage, adaptability), and e) three regarding the implementation process (e.g., integration into existing structures). Furthermore, we found strong interrelations between several of the influencing factors within and between domains. This comprehensive process evaluation complements the outcome evaluation of the SDM implementation trial and adds to its interpretation. The identified influencing factors can be used for planning, conducting, and evaluating SDM implementation in the future. clinicaltrials.gov, NCT03393351, registered 8 January 2018, https://clinicaltrials.gov/ct2/show/NCT03393351.


Correlation between aortic valve protein levels and vector flow mapping of wall shear stress and oscillatory shear index in patients supported with continuous-flow left ventricular assist devices.
Continuous-flow left ventricular assist devices commonly lead to aortic regurgitation, which results in decreased pump efficiency and worsening heart failure. We hypothesized that non-physiological wall shear stress and oscillatory shear index alter the abundance of structural proteins in aortic valves of left ventricular assist device (LVAD) patients. Doppler images of aortic valves of patients undergoing heart transplants were obtained. Eight patients had been supported with LVADs, whereas 10 were not. Aortic valve tissue was collected and protein levels were analyzed using mass spectrometry. Echocardiographic images were analyzed and wall shear stress and oscillatory shear index were calculated. The relationship between normalized levels of individual proteins and in vivo echocardiographic measurements was evaluated. Of the 57 proteins of interest, there was a strong negative correlation between levels of 15 proteins and the wall shear stress (R < -0.500, p ≤ 0.05), and a moderate negative correlation between 16 proteins and wall shear stress (R -0.500 to -0.300, p ≤ 0.05). Gene ontology analysis demonstrated clusters of proteins involved in cellular structure. Proteins negatively correlated with WSS included those with cytoskeletal, actin/myosin, cell-cell junction and extracellular functions. C: In aortic valve tissue, 31 proteins were identified involved in cellular structure and extracellular junctions with a negative correlation between their levels and wall shear stress. These findings suggest an association between the forces acting on the aortic valve (AV) and leaflet protein abundance, and may form a mechanical basis for the increased risk of aortic leaflet degeneration in LVAD patients.


Isolation and properties of chum salmon prolactin.
A highly purified prolactin (PRL) was isolated from the chum salmon pituitary by extraction with acid acetone, gel filtration on Sephadex G-25 and ion-exchange chromatography on CM-Sephadex C-25 with a yield of 1 mg/g of wet tissue. It was 10-15 times more potent than ovine PRL in sodium-retaining activity for juvenile rainbow trout adapted to 50% seawater. The salmon PRL emerged as a single and symmetrical peak on Sephadex G-100 with Ve/Vo = 2.0. Polyacrylamide gel electrophoresis revealed only one band at pH 4.3, whereas no band was seen at pH 7.5. The isoelectric point was estimated to be 10.3 by gel electric focusing. The circular dichroism spectrum of the salmon PRL was similar to that of tilapia PRL, showing an alpha-helix content of 50%. The salmon PRL had a molecular weight of 23,400 daltons by gel filtration and 22,300 daltons by sodium dodecyl sulfate gel electrophoresis, with a single NH2-terminal residue, isoleucine, and a single COOH-terminal residue, half-cystine. In the sequence comparison with those of mammalian PRLs and growth hormones, the clusters of invariant residues were found in both terminal regions, although the disulfide at NH2-terminal of mammalian PRLs was missing. Specific salmon PRL antisera were prepared in rabbits giving a precipitin reaction against the salmon PRL and a pituitary extract of tilapia in agar diffusion but no cross reaction with purified mammalian PRLs. The antibody was localized specifically in PRL cells of the chum salmon pituitary.


Preface - Emerging Viruses: From Early Detection to Intervention.
In the last decades, several viruses emerged, as the consequence of cross-species passage from animal reservoirs to human being. Emerging infections always represent a significant challenge for public health system as knowledge about specific pathogens are generally little while and no/few effective interventions are available. In this volume we explored special aspects of emerging infectious diseases including: the application of the theory of focality of diseases to infective syndrome; the human-animal inter-face focusing on influenza; the role of bats as main reservoir of emerging and novel human pathogens; Middle East Respiratory Syndrome Coronavirus (MERS-CoV) epidemiology and clinical presentation; immediate impact on human health of the last Zika virus outbreak occurred in Pacific Ocean and the Americas; the role and potential application of animal models for the study of emerging infections, to improve disease knowledge and for developing therapeutic drugs during ongoing outbreak.


Prenatal diagnosis, associated findings and postnatal outcome in fetuses with congenitally corrected transposition of the great arteries.
To analyze anatomic features and associated malformations in 37 prenatally detected cases of congenitally corrected transposition of the great arteries (ccTGA) and to evaluate the prenatal course, neonatal outcome and mid-term follow-up. Retrospective analysis of prenatal ultrasound of 37 patients with ccTGA in two tertiary centers between 1999 and 2019. All fetuses received fetal echocardiography and a detailed anomaly scan. Postnatal outcome and follow-up data were retrieved from pediatric reports. Isolated ccTGA without associated cardiac anomalies was found in 13.5% (5/37), in all other fetuses additional defects such as VSD (73.0%), pulmonary obstruction (35.1%), tricuspid valve anomalies (18.9%), aortic arch anomalies (13.5%), ventricular hypoplasia (5.4%) or atrioventricular block (5.4%) were present. The rate of extracardiac malformations or chromosomal aberrations was low. There were 91.9% (34/37) live births and postnatal survival rates reached 91.2% in a mean follow-up time of 4.98 years. The prenatal diagnosis of ccTGA was confirmed postnatally in all but one documented live birth and the prenatal counselling regarding the expected treatment after birth (uni- versus biventricular repair) was reassured in the majority of cases. The postnatal intervention rate was high, 64.7% (22/34) received surgery, the intervention-free survival was 36.7%, 35.0% and 25.0% at 1 month, 1 year and 10 years, respectively. ccTGA is a rare heart defect often associated with additional heterogeneous cardiac anomalies that can be diagnosed prenatally. The presented study demonstrates a favorable outcome in most cases but the majority of patients require surgical treatment early in life.


Correction to: Safety of psychotropic medications in people with COVID-19: evidence review and practical recommendations.
An amendment to this paper has been published and can be accessed via the original article.


Tetanus immunization shortage in the United States.
The purpose of this study is to determine the effect of the tetanus immunization shortage on EDs and the EPs understanding of the prioritization of persons needing tetanus immunization. A survey consisting of questions about knowledge of the tetanus shortage, prioritization of immunizations, incidence of tetanus infections, and understanding of CDC recommendations was mailed to a random sample 20% of the US ED medical directors. The results of the survey were input into the SPSS program (SPSS, version 10, Chicago, IL). The survey was returned by 618 of the 1,375 (44.9%) ED medical directors in the United States. Almost all (97.2%, 601 of 617) were notified about the tetanus shortage and 58.3% (360 of 617) reported a shortage. A total of 42.2% (199 of 472) gave tetanus toxoid (TT), instead of tetanus and diphtheria toxoids adult type (dT) when indicated. Only 11.6% of those surveyed (56 of 482) established a patient callback system. Routine vaccination was stopped in 37.5% of the reporting hospitals, most often for adults and children (57.5%, 69 of 120). Twelve hospitals (1.9%) reported they had an increase in tetanus. Although 87.5% of the respondents (539 of 616) stated they were familiar with the CDC's prioritization for tetanus immunization, only 1.8% (11 of 616) got the prioritization correct. Although EM directors uniformly know about and are experiencing the tetanus shortage, few correctly reported the tetanus immunization priority. Few EDs had a patient callback system.


Penile ecthyma gangrenosum. Complication of drug addiction.
We present a case of ecthyma gangrenosum involving the penis in an IV drug abuser probably resulting from direct arterial septic embolization from a femoral injection site. An increased awareness of this condition is essential because misdiagnosis has been reported in up to 100 percent of cases. Prompt diagnosis is essential to begin appropriate therapy.


Development of a DDH Care Pathway for India: A Study Methodology to Guide Similar Efforts in Other Countries and for Other Conditions.
In India and other Global South countries, developmental dysplasia of the hip (DDH) is often diagnosed after walking age, leading to more invasive surgeries and long-term disability. DDH care pathways aim to enhance early detection and must be tailored to meet a country's needs and diverse practice settings. We describe a multi-phase methodology for context-specific DDH care pathway development, demonstrating its use in India. In Phase I, Orthopaedic surgeons, Pediatricians/Neonatologists, and Radiologists in India were surveyed regarding DDH screening. Seven relevant Indian organizations partnered together and assembled a multidisciplinary working group, which then met fortnightly to establish an evidence base and prepare for the subsequent consensus-building phase. During Phase II, panelists participated in a modified Delphi process to reach consensus on a list of DDH screening statements. Phase III applied the statements to develop the care pathway. The Delphi process concluded after a preliminary survey and two Delphi rounds, reaching consensus on 47 statements, which were condensed into 35. The developed care pathway for India features periodic clinical hip examinations integrated with the country's immunization schedule and selective imaging screening, providing flexibility in the timing and modality of imaging. In Global South countries, there is a need for DDH care pathways specific to local contexts. Successful care pathway development requires accounting for cultural differences in healthcare and strategies to facilitate engagement and to address country-specific barriers. This methodology was feasible in India and can be applied to other conditions and/or countries wishing to establish care pathways. Level III.


Facial Rejuvenation and Contouring with Radiofrequency-Assisted Procedures in Asians.
Radiofrequency (RF)-assisted liposuction treatment is a minimally invasive skin-tightening technique for the patient population with skin laxity. The authors recommend facial liposuction combined with the RF procedure for the treatment of skin laxity. Minimal-invasive liposuction creates working channels for RF treatment and sufficiently exposes the subdermal fibrous septal network tissue so that the RF energy can directly act on the collagen of the fibrous septal network for thermal shrinkage, leading to better surgical results. In this article, the authors describe their preferred technique and experience for face rejuvenation and contouring.


Laparoscopic monolateral suspension for vaginal vault prolapse: a report of an exit surgical strategy during sacralcolpopexy.
Vaginal vault prolapse is the most frequent long-term complication in patients undergoing hysterectomy and sacralcolpopexy is considered the gold standard. We report our surgical strategy maintaining single-arm mesh when the sacral promontory is not accessible to fix the mesh for an unknown sacral osteophytosis during a laparoscopic sacralcolpopexy. This is significant because, to our knowledge, the bone variant as a procedure limiting factor has never been described before. This opens new horizons for the sacralcolpopexy surgery, because it becomes necessary to know of a valid surgical alternative with mesh maintenance if this complication occurs again or to perform an assessment of the accessibility of the sacral promontory immediately after its dissection. We present a case of a 75-year-old woman with recurrence of vaginal vault prolapse. A laparoscopic sacralcolpopexy was recommended. During surgery, we found that the procedure was not feasible due to the presence of an unknown osteophytosis of the sacrum which prevented the fixing of the mesh to the sacral promontory. We decided to proceed with a single-arm lateral suspension by using a modified approach of the original technique, maintaining the mesh originally shaped for the sacral colpopexy. At follow-up, the vaginal vault is well suspended. This exit strategy may represent a valid surgical alternative when laparoscopic sacral colpopexy is not possible for anatomical variants, allowing to keep the laparoscopic approach using mesh. To our knowledge, cases in which the anatomical bone variant prevented access to the sacral promontory have never been described in the literature, as bone evaluation has never been considered a limiting element of this procedure.


Specific PCR assays to determine bovine, porcine, fish and plant origin of gelatin capsules of dietary supplements.
Gelatin, a purified protein derived mostly from pig skin and bovine tissue, is used widely in both food and pharmaceutical industries. Here, to determine the species of origin of capsule gelatin, we developed a sensitive and reliable test using the polymerase chain reaction (PCR) method, which included 1) species-specific or universal primer sets, designed to detect short 16S ribosomal RNA (rRNA) gene sequences from cow, pig, and fish (tilapia) as well as genes encoding the large subunit of plant ribulose-1,5-bisphosphate carboxylase oxygenase and 2) species-specific PCR coupled with whole-genome amplification. This method was used to verify manufacturing label claims of 28 gelatin capsule samples sold as dietary supplements. The results from 27 samples were consistent with gelatin-related information on the manufacturer label, while one sample that mentioned tilapia gelatin was found to contain only bovine DNA. This rapid method can therefore be used to verify the authenticity of gelatin capsules.


Local blood flow regulation in normotensive man during prolonged treatment with propranolol.
The influence of propranolol on blood flow in peripheral tissues was studied in 10 normotensive subjects. Adequacy of beta-blockade was ascertained by an ergometric test. Perfusion of the subcutaneous tissue and skeletal muscle was measured by the local 133Xe-washout technique. The arteriolar dilatation underlying the autoregulatory response to a decrease in arterial perfusion pressurehead was not affected by beta-blockade. Propranolol did not change the magnitude of arteriolar constriction elicited by the local sympathetic veno-arteriolar reflex. Thus, the locally induced increase in the discharge rate along the peripheral sympathetic adrenergic fibres did not lead to measurable change in beta-receptor activity. The present results indicate that local bloodflow regulatory mechanisms are not affected by propranolol. They also indicate that the direct peripheral effects of propranolol are of no clinical importance in normotensive subjects in the supine position, when the general activity in the sympathetic nervous system is low.


Duty cycle and modulation efficiency of two-channel Hadamard transform time-of-flight mass spectrometry.
Hadamard transform time-of-flight mass spectrometry (HT-TOFMS) is based on the pseudorandom gating of ion packets into a time-of-flight mass-to-charge analyzer. In its typical implementation, the technique is able to monitor continuous ion sources with a 50% duty cycle, independent of all other figures of merit. Recently, we have demonstrated that the duty cycle can be extended to 100% using patterned, two-channel detection. Two-channel HT-TOFMS involves the simultaneous optimization of paired one-channel experiments and imposes more stringent conditions to achieve high-quality spectra. An ion modulation device, known as Bradbury-Nielson Gate (BNG), is central to HT-TOFMS. It is an ideal deflection plate, capable of transmitting or deflecting an ion beam according to a known binary sequence without changing the times-of-flight of the ions. Analytical equations are derived that accurately describe the ion modulation process of the BNG as confirmed by good agreement with SimIon simulations and ion beam imaging experiments. From these expressions, the duty cycle and ion modulation efficiency were calculated for various BNG parameters, ion beam characteristics, and detector dimensions, which permit the optimum conditions to be chosen for the two-channel experiment. We conclude that the outer detector should be three times the maximum deflection angle to detect all deflected ions (100% duty cycle) and that the difference between the modulated ion counts in the sequence elements 0 and 1 should be maximized to achieve high modulation efficiency. This condition is best achieved by tight focusing of the ion beam in the center of the inner detector. When both channels are optimized, the two-channel advantage can be exploited to achieve a further improvement over a single-channel experiment.


Extinction and the temporal distribution of macroevolutionary bursts.
Phenotypic evolution through deep time is slower than expected from microevolutionary rates. This is the paradox of stasis. Previous models suggest stasis occurs because populations track adaptive peaks that remain relatively stable on million-year intervals, raising the equally perplexing question of why these large changes are so rare. Here, we consider the possibility that peaks can move more rapidly than populations can adapt, resulting in extinction. We model peak movement with explicit population dynamics, parameterized with published microevolutionary estimates. Allowing extinction greatly increases the parameter space of peak movements that yield the appearance of stasis observed in real data through deep time. Extreme peak displacements, regardless of their frequency, will rarely result in an equivalent degree of trait evolution because of extinction. Thus, larger peak displacements will rarely be inferred using trait data from extant species or observed in fossil records. Our work highlights population ecology as an important contributor to macroevolutionary dynamics, presenting an alternative perspective on the paradox of stasis, where apparent constraint on phenotypic evolution in deep time reflects our restricted view of the subset of earth's lineages that were fortunate enough to reside on relatively stable peaks.


Linear and nonlinear systems analysis of the visual system: why does it seem so linear? A review dedicated to the memory of Henk Spekreijse.
Linear and nonlinear systems analysis are tools that can be used to study communication systems like the visual system. The first step of systems analysis often is to test whether or not the system is linear. Retinal pathways are surprisingly linear, and some neurons in the visual cortex also emulate linear sensory transducers. We conclude that the retinal linearity depends on specialized ribbon synapses while cortical linearity is the result of balanced excitatory and inhibitory synaptic interactions.


Self-Assembly of 3D DNA Crystals Containing a Torsionally Stressed Component.
There is an increasing appreciation for structural diversity of DNA that is of interest to both DNA nanotechnology and basic biology. Here, we have explored how DNA responds to torsional stress by building on a previously reported two-turn DNA tensegrity triangle and demonstrating that we could introduce an extra nucleotide pair (np) into the original sequence without affecting assembly and crystallization. The extra np imposes a significant torsional stress, which is accommodated by global changes throughout the B-DNA duplex and the DNA lattice. The work reveals a near-atomic structure of naked DNA under a torsional stress of approximately 14%, and thus provides an example of DNA distortions that occur without a requirement for either an external energy source or the free energy available from protein or drug binding.


Techniques for acute single neuron recording in newborn rats.
Techniques are described for artifically respiring, anesthetizing, head-holding, operating on and recording from single neurons in the VIII nerve of newborn rats weighting as little as 4.9 g.


Penile enhancement using a porcine small intestinal submucosa graft in a rat model.
Several biodegradable materials have been experimented for penile enhancement, but none show the potential for clinical use. This study was designed to use porcine small intestinal submucosa (SIS) augmenting the normal tunica albuginea to increase the functional girth of the rat penis. In all, 20 adult male Sprague-Dawley rats constituted the study population. The animals were divided into two groups: group 1 consisted of the control (n=10) and group 2 (n=10) consisted of rats that underwent penile enhancement by a longitudinal I-shaped incision of the tunica albuginea on both sides, and the dissection of the plane between tunica albuginea and cavernosal tissue was carried out (n=10). The incision was then patched with a 3 x 10 mm2 piece of SIS, using a 6/0 nylon suture material. The penile length and mid-circumference were then measured using a Vernier Caliper before and 2 months after surgery. All rat penises underwent histological examination using Masson's trichome and Verhoff's van Giesen's stain for collagen and elastic fibers. The penile length, mid-circumference and degree of fibrosis score were expressed as mean+/-s.e. (standard error) and analyzed using a Wilcoxon rank-sum test. A statistical significance was accepted at P-value < or =0.05. Our results showed similar preoperative penile length and circumference in both groups. However, 2 months after the surgery, the mean penile circumference of the SIS group has grown significantly larger than the control group, while the mean penile length remained unchanged. The histological study of the rat penises revealed minimal amounts of fibrosis under the graft, and the elastic fibers of the graft showed orientation in a circular manner. In conclusion, SIS appears promising for material use in a penile enhancement.


Habitual level of physical activity and cardiorespiratory endurance capacity in children.
A total of 257 healthy children (140 boys, 117 girls) varying in age from 5.7 to 18.5 years underwent graded exercise tests on a treadmill. Cardiorespiratory endurance capacity was assessed by determination of the ventilatory threshold, which was defined as the highest exercise intensity before a disproportionate increase occurred in pulmonary ventilation (VE) relative to oxygen uptake (VO2). The purpose of the present study was to investigate the relationship between the habitual level of physical activity (HLPA) and the cardiorespiratory endurance capacity in children. The HLPA was assessed by a standardized questionnaire. In boys and girls HLPA increased gradually with advancing age. For the group as a whole, the boys reached the highest values for HLPA. The most active boys reached the highest value for ventilatory threshold and the lowest value was found in the less active ones, except for the age span of 12-16 years. It is concluded that more active boys showed a higher cardiorespiratory endurance capacity, as assessed by the ventilatory threshold, compared to less active ones, except during puberty. It can be postulated that during puberty this effect may be overruled by the influence of other more dominant growth-related factors. In girls, because HLPA was rather low, there was no discriminative effect of HLPA on exercise performance, as would be expected.


Effects of TCDD-estradiol interactions in three strains of mice.
Interactions of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and estradiol were studied in three strains of mice: CD-1 and C57B/6 (TCDD sensitive) and DBA/2 (TCDD resistant at lower doses). Immature females were injected with 0-200 ng/kg/day estradiol for 2 weeks, s.c. On days 7, 9, 11, and 13, mice received 10 micrograms TCDD/kg by gavage. Relative uterine weight increased in mice of all three strains treated with estradiol alone. Uterine imbibition was suppressed by TCDD treatment, although this effect was antagonized in a saturable manner by coadministration of estradiol. TCDD induced aryl hydrocarbon hydroxylase (AHH) in liver microsomes of treated mice independent of estradiol dose and strain of mice (the dose of TCDD used was high enough to cause AHH induction in the resistant DBA/2 mice). Treatment of CD-1 mice, but not other strains, with TCDD and estradiol resulted in a decrease in the electrophoretic band of hepatic microsomal proteins comigrating with cytochrome P-450a and epoxide hydrase.


Epidemiological aspects of spontaneous subarachnoid haemorrhage in Queensland, Australia.
To examine the epidemiology of spontaneous subarachnoid haemorrhage (SAH) within the population of Queensland, Australia in 2002. A retrospective population and hospital-based survey of all cases of spontaneous SAH occurring within the population of Queensland (3.7 million) during the calendar year 2002 was performed. Cases were identified from hospital separation coding data and the register of births, deaths and marriages. Standard demographic data was recorded for each case identified. The annual incidence of SAH in our population was 9.4 cases per 100,000. There was a steady increase in the incidence of SAH with increasing age, with the incidence rising to 38.8 per 100,000 in those aged greater than 80. The overall mortality rate was 33.1%, with 6% of all cases dying before reaching hospital care. The annual incidence in the indigenous population of Queensland was 8.9 /100,000.


Bioassay-guided isolation of an alkaloid with antiangiogenic and antitumor activities from the extract of Fissistigma cavaleriei root.
Fissistigma cavaleriei (Levl) Rehd (Annonaceae) is used as a folklore medicine for treatment of inflammation, arthritis, and tuberculosis by Miao people in China. In the present study, the antiangiogenic activity of F. cavaleriei was investigated. The chorioallantoic membrane of the fertilized hen's egg (CAM assay) was used to determine antiangiogenic activity of the plant extract. Compound (1), a compound with antiangiogenic activity, was isolated by bioassay-guided fractionation from F. cavaleriei for the first time. The structure of compound (1) was elucidated on the basis of spectroscopic methods. Colorimetric COX (ovine) inhibitor screening assay was used to determine its inhibitory effect on COX-1 and COX-2. MTT and Sulforhodamine B assays were used to investigate its cytotoxic effects on tumor cell lines. As a result, compound (1) showed a selectively inhibiting effect on COX-2 and could inhibit the growth of tumor cells in vitro. The antitumor activity of compound (1) was further confirmed by the observation that compound (1) administration significantly inhibited the growth of S-180 cells in mice. Moreover, compound (1) was able to enhance the antitumor activity of doxorubicin in the mice bearing with S-180 cells while combined with doxorubicin. In conclusion, compound (1) is a multi-target molecule and further experimental investigations are needed to determine whether it can be used as a lead molecule for tumor treatment.


Chondroptosis: an immunohistochemical study of apoptosis and Golgi complex in chondrocytes from human osteoarthritic cartilage.
The Golgi complex is thought to play an important role in the apoptotic process of osteoarthritic (OA) chondrocytes. However, the exact relationship between modifications of the Golgi complex and apoptosis in human OA cartilage requires to be established. We compared the patterns and immunolabeling intensities for anti-Golgi 58 K protein with apoptosis markers such as TUNEL and caspase-2L in OA cartilage removed from patients during knee total replacement surgery. We observed important modifications in labeling of the Golgi 58 K protein in OA chondrocytes compared with normal cell. Immunohistochemical analysis revealed co-localization between 58 K protein and caspase-2L, suggesting that this enzyme was localized in Golgi complex of OA chondrocytes. In addition, these cells labeled positive with the TUNEL technique, but in different proportions to caspase-2L. Our results support the concept, previously reported, that apoptosis in OA cartilage (chondroptosis) might be a variant of the classical apoptosis.


Workflow and decision making of operating room nurses: integrative review.
Identify workflow factors in the operating room and their implications, which influence nurses' decision making. Integrative review of the literature conducted through searches in the databases: Latin American and Caribbean Literature in Health Sciences; Nursing Database; Pubmed; Scopus and Cumulative Index to Nursing and Allied Health Literature. The results were organized into factors related to positive, negative and positive and negative implications. The sample of 18 articles included examples of factors with positive implications, such as preoperative data collection, negative outcomes, such as lack of human, material and structural resources, and positive and negative outcomes, as preparation for certification. Factors that influence the decision-making process of nurses are associated to different conditions: client- related conditions and those conditions that go beyond the domain and organization of the surgical environment.


Cortical Microinfarcts Associated With Worse Outcomes in Patients With Acute Ischemic Stroke Receiving Endovascular Treatment.
We aimed to evaluate the impact of cortical microinfarcts (CMIs) on functional outcome after endovascular treatment in patients with acute ischemic stroke. In a multicenter registration study for RESCUE-RE (a registration study for Critical Care of Acute Ischemic Stroke After Recanalization), eligible patients with large vessel occlusion stroke receiving endovascular treatment, who had undergone 3T magnetic resonance imaging on admission or within 24 hours after endovascular treatment were analyzed. We evaluated the presence and numbers of CMIs with assessment of axial T1, T2-weighted images, and fluid-attenuated inversion recovery images. The primary outcome was functional dependence or death defined as modified Rankin Scale scores of 3 to 6 at 90 days. Secondary outcomes included early neurological improvement, any intracranial hemorrhage, symptomatic intracranial hemorrhage, and mortality. We investigated the independent associations of CMIs with the outcomes using multivariable logistic regression in overall patients and in subgroups. Among 414 patients (enrolled from July 2018 to May 2019) included in the analyses, 96 (23.2%) patients had at least one CMI (maximum 6). Patients with CMI(s) were more likely to be functionally dependent or dead at 90 days, compared with those without (55.2% versus 37.4%; P<0.01). In multivariable logistic regression analyses, presence of CMI(s) (adjusted odds ratio, 1.78 [95% CI, 1.04-3.07]; P=0.04) and multiple CMIs (CMIs ≥2; adjusted odds ratio, 7.41 [95% CI, 2.48-22.17]; P<0.001) were independently, significantly associated with the primary outcome. There was no significant difference between subgroups in the associations between CMI presence and the primary outcome. Acute large vessel occlusion stroke patients receiving endovascular treatment with CMI(s) were more likely to have a poor functional outcome at 90 days, independent of patients' characteristics. Such associations may be dose-dependent. Registration: URL: http://www.chictr.org.cn; Unique identifier: ChiCTR1900022154.


Prevalence of IgG and IgM antibodies specific to Toxoplasma gondii in cats.
IgG and IgM antibodies specific to Toxoplasma gondii were determined simultaneously by IFAT in cats. Group 1 included 357 cats with various diagnoses, but negative for FeLV and FIV. Specific IgG antibodies in titres ranging from 10 to 2560 (mean 247) were detected in 61.3% animals, specific IgM antibodies were found only in one cat (0.28%) in titre 1:40 showing an IgG titre of 160. Oocyst shedding was demonstrated in one cat (0.28%) showing an IgG titre of 1:40. Group 2 included 33 cats positive for FeLV or FIV. IgG antibodies in titres ranging from 10 to 640 (mean 101) were found in 63.6% cats. Neither IgM antibodies nor shedding of T. gondii oocysts were detectable. Clinically unapparent toxoplasmosis, manifested by the presence of IgG antibodies is frequent in cats; the opportunistic character of the infection is important.


Morphological and molecular characterization of Thelohanellus macrovacuolaris n. sp. (Myxosporea: Bivalvulida) infecting the palate in the mouth of common carp Cyprinus carpio L. in China.
Thelohanellus macrovacuolaris n. sp. is described during a survey on myxozoan diversity of common carp Cyprinus carpio L. in China. It is characterized by the presence of round or ellipsoidal plasmodium in the palate in the mouth of host. Mature spores were pyriform in frontal view, lemon shaped in lateral view, measuring 21.6±0.9 (19.3-23.8) long, 12.5±0.7 (10.3-13.6) wide, and 10.2±0.4 (9.8-11.8) thick. Most spores were surrounded by the membrane sheath. Single polar capsule was round with an apophysis at its top end presented close to apex of spore, measuring 9.1±0.6 (8.0-10.0) in length, 8.6±0.5 (7.8-9.6) in width. Polar filaments coiled, with 7 to 8 turns. A large, round iodinophilous vacuole was present, with 5.8-7.5 in diameter. The present species is morphologically distinct from all other Thelohanellus species. The BLAST search indicated that the newly obtained small subunit ribosomal RNA (ssrRNA) gene sequence of T. macrovacuolaris n. sp. did not match any available sequences in GenBank. Phylogenetically, T. macrovacuolaris n. sp. was placed sister to Thelohanellus wangi in the Thelohanellus clade. Both morphology and ssrRNA gene sequence data revealed that the present parasite is a new species of genus Thelohanellus.


In vivo acute physiological stress induced by HCH on the levels of glycogen and total carbohydrates in various tissues of Oziotelphusa senex senex, the Indian ricefield crab.
The ricefield crab (Oziotelphusa), when exposed to a lethal concentration (LC50/24 h = 39 mg/l) of hexachlorocyclohexane (HCH) to study acute toxic effects, revealed a significant (P less than 0.001) decrease in the content of total carbohydrates (TCHO) and glycogen of the hepatopancreas and muscle. No significant changes were observed in TCHO and glycogen levels of the gonad, hypodermis and gill.


Using Citizen Science to Explore Neighbourhood Influences on Ageing Well: Pilot Project.
Outdoor and indoor environments impact older people's mobility, independence, quality of life, and ability to "age in place". Considerable evidence suggests that not only the amount, but also the quality, of public green spaces in the living environment is important. The quality of public green spaces is mostly measured through expert assessments by planners, designers and developers. A disadvantage of this expert-determined approach is that it often does not consider the appraisals or perceptions of residents. Daily experience, often over long periods of time, means older residents have acquired insider knowledge of their neighbourhood, and thus, may be more qualified to assess these spaces, including measuring what makes a valued or quality public green space. The aim of this Australian pilot study on public green spaces for ageing well was to test an innovative citizen science approach to data collection using smart phones. "Senior" citizen scientists trialed the smart phone audit tool over a three-month period, recording and auditing public green spaces in their neighbourhoods. Data collected included geocoded location data, photographs, and qualitative comments along with survey data. While citizen science research is already well established in the natural sciences, it remains underutilised in the social sciences. This paper focuses on the use of citizen science with older participants highlighting the potential for this methodology in the fields of environmental gerontology, urban planning and landscape architecture.


Differential and dose-dependent regulation of gene expression at the mid-hindbrain boundary by Ras-MAP kinase signaling.
Recent experiments suggest that activation of the Ras-MAP kinase pathway at the mid-hindbrain boundary (MHB) induces cerebellar development, whereas tectal development occurs in the absence of Ras-MAP kinase activity. To test this model we have stimulated or inhibited Ras-MAP kinase signaling in chick embryos through targeted misexpression of a constitutive active (Ras(V12)) or dominant negative (Ras(N17)) form of Ras. The consequence of these manipulations on the expression of several genes that are expressed in distinct patterns at or around the MHB organizer, including En1, Pax2, Pax3, Pax5, Wnt1, Meis2, and ephrin-A2, -A5, and -B1, was assessed. Extending previous findings we show that inhibition of Ras-MAP kinase signaling differently affects Pax3 expression in different regions of the mid-hindbrain territory, inhibiting its expression in the midbrain but inducing it in the MHB region. Expression of the midbrain specific marker gene Meis2 was not affected by Ras(N17) at first but later upregulated concomitantly with the morphological transformation of hindbrain to midbrain. In addition, we show that different dosages of Ras-MAP kinase activity are required for transcriptional activation of Wnt1 and En1 at the MHB. Collectively, these results validate and extend previous findings on the molecular changes associated with Fgf8 loss-of-function or gain-of-function phenotypes at the MHB, demonstrate that gene expression at the MHB is regulated by Ras-MAP kinase signaling in a spatially and temporally distinct manner and provide evidence for a dosage dependent function of Fgf8 signaling at the MHB.


HIV rapid test performance among health facilities enrolled in HIV rapid test quality improvement initiative (RTQII) in Ethiopia.
As the Human Immunodeficiency Virus (HIV) rapid testing services expanded to reach the global target that 95% of people living with the virus will know their status by 2030, ensuring the quality of those services becomes critical. This study was conducted to assess the performance of HIV Rapid testing at sites in health facilities that were enrolled in the Rapid Test Quality Improvement Initiative (RTQII) in Ethiopia. Characterized HIV proficiency testing (PT) panels of Dried Tube Specimen (DTS) were prepared, verified, and distributed to testing sites from August to December 2019. In addition on-site evaluation of HIV testing sites (HTSs) was conducted using a checklist to assess testing conditions. For proficiency testing, the study included 159 HIV testing sites (HTSs) in 41 Health facilities (HFs) in five administrative regions and two city administrations. The collected data was analyzed by SPSS version 20 and chi-square test was applied to identify the association between acceptable performance and contributing factors. Testing sites with 100% PT score as well as conducting the test with adherence to the National HIV Testing Algorithm were considered acceptable. The overall acceptable performance (100% PT score with the correct algorithm followed) was found to be 62% while 12% scored 80% and 11% scored between 20 and 60%. The rest 15% were not considered as acceptable due to failure to adhere to the National HIV Testing Algorithm. Testing sites that participated in External Quality Assessment/Proficiency Testing schemes have shown better performance than those that did not participate with 70% and 56% performance respectively (p = 0.057).


Mechanisms of CD8+ T cell exclusion and dysfunction in cancer resistance to anti-PD-(L)1.
CD8+ T cells are the front-line defensive cells against cancer. Reduced infiltration and effector function of CD8+ T cells occurs in cancer and is contributed to defective immunity and immunotherapy resistance. Exclusion and exhaustion of CD8+ T cells are the two key factors associated with reduced durability of immune checkpoint inhibitor (ICI) therapy. Initially activated T cells upon exposure to chronic antigen stimulation or immunosuppressive tumor microenvironment (TME) acquire a hyporesponsive state that progressively lose their effector function. Thus, a key strategy in cancer immunotherapy is to look for factors contributed to defective CD8+ T cell infiltration and function. Targeting such factors can define a promising supplementary approach in patients receiving anti-programmed death-1 receptor (PD-1)/anti-programmed death-ligand 1 (PD-L1) therapy. Recently, bispecific antibodies are developed against PD-(L)1 and a dominant factor within TME, representing higher safety profile and exerting more desired outcomes. The focus of this review is to discuss about promoters of deficient infiltration and effector function of CD8+ T cells and their addressing in cancer ICI therapy.


Multidimensional NMR spectroscopy for protein characterization and assignment inside cells.
High-field, heteronuclear NMR spectroscopy of biological macromolecules in native cellular environments is limited by the low concentrations present and the long data acquisition times needed for the experiments. Successful 1D and 2D heteronuclear NMR data have been reported, but the 3D experiments conventionally used for protein assignment and detailed characterization are generally too long to maintain cell viability. Here we describe the successful in vivo implementation of a suite of fast 3D NMR experiments which we have used to generate the complete backbone assignment of resonances in the recombinant polypeptide GB-1 within Escherichia coli cells. The data were acquired at 600 MHz with a cold probe using the projection reconstruction experiments, (3,2)HNCA, (3,2)HNCO, and (3,2)HA(CA)NH.


Increased Serum Cu/Zn SOD in Individuals with Clinical Depression Normalizes After Zinc and Anti-oxidant Therapy.
To assess serum Cu/Zn SOD (Superoxide Dismutase) concentration in individuals with clinical depression. Serum from 36 individuals diagnosed with clinical depression and 18 healthy controls were tested for Cu/Zn SOD serum concentration using ELISAs. Serum Cu/Zn SOD levels of depressed individuals (both with and without secondary anxiety) were significantly higher than age and gender similar controls. We also found that, post anti-oxidant therapy, Cu/Zn SOD levels normalized to the level of normal healthy controls. These results suggest an association between Cu/Zn SOD serum levels and clinical depression.


[Heat production in the early stages of axolotl growth according to direct and indirect calorimetry findings].
The level of energetic metabolism of the growing axolotls was studied by the methods of direct and indirect calorimetry. Just after the hatching the heat production of the larvae calculated by the gas exchange exceeds markedly the level of heat production measured by the calorimeter. During the subsequent growth the level of heat production changes as a transitional process, so that therafter indirect calorimetry gives constantly higher values of energetic metabolism than direct one.


Pyroptosis executor gasdermin D plays a key role in scleroderma and bleomycin-induced skin fibrosis.
The NLRP3 inflammasome and IL-1β are essential for scleroderma pathogenesis. Nevertheless, the role of pyroptosis executor gasdermin D(GSDMD), which is a downstream molecule of NLRP3 and is required for IL-1β release in some situations, has not yet been well elucidated in scleroderma. Here, we found that GSDMD was significantly up-regulated and activated in the skin of scleroderma patients and bleomycin-induced mouse model. What's more, the ablation of GSDMD ameliorates bleomycin-induced skin fibrosis according to HE staining, Masson staining and the detection of hydroxyproline contents. GSDMD deficiency also impaired macrophages infiltration and reduced inflammation response. Furthermore, the loss of GSDMD reduced Th17 differentiation in vivo and in vitro. Collectively, these findings provide the first demonstration that GSDMD related pyroptosis plays an important role in scleroderma pathogenesis.


Rating of Perceived Exertion associated with acute symptoms in athletes with recent SARS-CoV-2 infection: AWARE VI.
SARS-CoV-2 infection can affect the exercise response in athletes. Factors associated with the exercise response have not been reported. This study aimed to: 1) describe heart rate (HR), systolic blood pressure (SBP), and rating of perceived exertion (RPE) responses to exercise in athletes with recent SARS-CoV-2 infection, and 2) identify factors affecting exercise responses. This was a cross-sectional, experimental study. Male and female athletes (age 24.2 ± 6.3 years) with recent (<28 days) SARS-CoV-2 infection (n=72). A 'COVID-19 Recovery Clinic' for athletes. HR, SBP and RPE were measured during submaximal exercise (Modified Bruce protocol) at 10-28 days after SARS-CoV-2 symptom onset. Selected factors (demographics, sport, co-morbidities, pre-infection training variables and symptoms during the acute phase of the infection) affecting the exercise response were analyzed using random coefficient (linear mixed) models. HR, SBP, and RPE increased progressively from rest to stage 5 of the exercise test (p=0.0001). At stage 5 (10.1 METs), a higher HR and SBP during exercise was associated with younger age (p=0.0007) and increased body mass index (BMI) (p=0.009) respectively. Higher RPE during exercise was significantly associated with greater number of whole body (p=0.006) and total number (p=0.004) of symptoms during the acute phase of infection. Greater number of symptoms during the acute infection was associated with a higher RPE during exercise in athletes with 10-28 days after SARS-CoV-2 infection. We recommend measuring RPE during the first exercise challenge after infection as this may indicate disease severity and be valuable to track progress, recovery, and return-to-sport.


Output-feedback asymptotic tracking control for rigid-body attitude via adaptive neural backstepping.
In this note, a novel neural adaptive output-feedback control (NAOC) with asymptotic tracking performance for rigid-body attitude is investigated subject to inertia uncertainty, unavailability of the angular velocity and unknown external disturbance. First, by virtue of the combination of the first-order filter and the coordinate transformation, the original output feedback system with immeasurable angular velocity and unknown dynamics is converted into the full-state strict feedback system with mismatched disturbance. Second, aided by infinitely integrable inequality with saturation function, an innovative neural network (NN) based adaptive control scheme is proposed via backstepping technique. By adopting the model transformation and proposed algorithm, the asymptotic tracking performance of the transformed system and the attitude tracking system without angular velocity can be achieved simultaneously. Finally, comparative numerical simulations illustrate the efficacy of the developed algorithm.


Epitope specificity and TCR V beta gene utilization in the encephalitogenic response of B10.RIII(71NS)/SnJ mice.
A high proportion of peptide 1-11 specific T cells from H-2u (V beta 8+, H-2u) mice express the V beta 8 TCR chain. Peptide 89-101 is immunodominant for B10.RIII (V beta 8+, H-2r) mice; thus, it was of interest to determine whether V beta 8 TCR would be over-represented in a population of peptide 89-101-specific T cells of this strain. Second, it was asked whether MBP peptides other than 89-101 would induce EAE in these mice. Of 70 B10.RIII(71NS)/SnJ mice immunized with mouse myelin basic protein (MBP), 32 of 41 males (78%) and 11 of 29 females (38%) showed clinical signs of experimental allergic encephalomyelitis (EAE). All mice immunized with peptide 89-101 showed clinical signs. One of six mice immunized with peptide 91-103 showed clinical signs, and 9 of 16 mice, all males, responded with EAE when immunized with peptide 38-88. No clinical EAE was observed in mice immunized with peptide 43-67, 68-88, 55-74, 1-37 or 1-20. A peptide 89-101-specific T cell line was established. At the initial stimulation the line was 29% V beta 8+ versus 21% in normal controls, and the line did not transfer EAE adoptively. After five in vitro stimulations, the percentage of V beta 8+ T cells had increased to 54%, and the line was encephalitogenic. Encephalitogenicity was partially blocked by anti-V beta 8 monoclonal antibody. Thus, over-representation of V beta 8+ TCR by encephalitogenic peptide-specific T cells is not limited to peptide 1-11-specific T cells from H-2u mice.(ABSTRACT TRUNCATED AT 250 WORDS)


Hepatitis A antibody persistence 8 and 10 years after 1-dose and 2-dose vaccination in children from Panama.
Hepatitis A virus (HAV) remains a global public health concern, which is potentially growing in Latin America, due to an expected shift from high to intermediate endemicity levels. The use of HAV vaccines in pediatric national immunization programs (NIPs), either as a 2-dose or a 1-dose schedule, has been explored in Latin American countries; however, evidence demonstrating long-term protection in this population is limited in the region. We evaluated long-term antibody persistence following a 1-dose partial series and the recommended 2-dose schedule used in Panama's pediatric NIP. Two independent cross-sectional serological surveys were conducted at year 8 (Y8) and Y10 following vaccination under the NIP with 1 or 2 doses of an inactivated HAV vaccine (Havrix, GSK). Seropositivity (anti-HAV antibody concentration ≥ 15 mIU/mL) rates and antibody geometric mean concentrations (GMCs) were assessed at each serosurvey. Non-inferiority of 1 dose versus 2 doses was also explored. This study (NCT02712359) included 600 and 599 children at Y8 and Y10 post-vaccination, respectively. Seropositivity rates were 74.3% (95% confidence interval [CI]: 69.0; 79.2) and 97.7% (95% CI: 95.3; 99.1) at Y8 and 71.9% (95% CI: 66.4; 76.9) and 96.3% (95% CI: 93.5; 98.2) at Y10, in the 1-dose and 2-dose groups, respectively. Antibody GMCs were lower in the 1-dose versus the 2-dose group in both surveys. Non-inferiority was not demonstrated since the lower limit of the 2-sided 95% CI for the between-group difference in seropositivity rates (1-dose minus 2-dose) was < -10%. Anti-HAV antibody persistence was observed in lower percentages of children receiving 1 dose versus 2 doses of Havrix, at 8 and 10 years post-vaccination in Panama. Further investigations are needed to confirm antibody persistence and conclude on the protection afforded beyond 10 years in the pediatric population in Latin America.


Determination of zolpidem in human hair by micropulverized extraction based on the evaluation of relative extraction efficiency of seven psychoactive drugs from an incurred human hair specimen.
A micropulverization method for rapid extraction of psychoactive drugs from hair was developed. A hair sample (10mg) was micropulverized for 10min at 42Hz with 0.2mL of 45% (w/v) aqueous ammonium phosphate (pH 8.4). Liquid-liquid extraction was carried out in the same tube using acetonitrile, and the organic layer was removed and filtered. Conventional methods, including solid-liquid extraction with an ammonium phosphate solution or methanol, were also employed, and the relative extraction efficiencies of amitriptyline, nortriptyline, norfludiazepam, flunitrazepam, 7-aminoflunitrazepam, mianserin and zolpidem with these methods from an incurred human hair specimen were compared using liquid chromatography/tandem mass spectrometry. The highest extraction efficiencies for all the analytes were achieved using the method developed here, even though the extraction time (10min) was short. Overnight methanol extraction has frequently been used for hair analysis; however, the extraction efficiency was not sufficient for amines. The method was successfully applied to the quantification of zolpidem in human hair. The range of quantification was 1-25,000pg/mg, and interday accuracy and precision (n=5) at three concentrations were 1.8-8.8% and 3.3-8.1%, respectively. The developed method was applied to three actual (incurred) samples, for which the concentrations of zolpidem were determined to be 78.9-18,300 (pg/mg).


To swim or not to swim? A disagreement between microbial indicators on beach water quality assessment in Hong Kong.
The USEPA and the WHO now advocate the use of enterococci as indicators for marine water quality. This study investigated the outcomes for Hong Kong beach water quality assessment by comparing enterococcus measures with data from the HKEPD's monitoring programme. Six beaches were tested once every 2-3 months from November 2013 to June 2014 in order to identify the most contaminated sites, followed by intensive water sampling in sites found to have the highest enterococci densities (Clear Water Bay Second and Golden) every five to six days for six sampling events over a 30-day period in 2014. The geometric means of enterococci were found to be 124 and 41 cfu/100 mL at Clear Water Bay Second and Golden respectively, indicating that there may be higher risks of illness associated with swimming at both beaches than previously known. Moreover, beach sediments contained higher concentrations of enterococci than water, and warrant further study.


Hematuria associated with cystic hematomas in three neonatal foals.
Cystic hematomas were diagnosed in 3 neonatal foals. In all 3 foals, cystic hematomas resulted from umbilical trauma or thrombocytopenia. Findings in these foals suggest that cystic hematoma should be included in the differential diagnosis of hematuria in neonatal foals. Management of cystic hematomas includes both medical and surgical options.


Adjacent segment stenosis after lumbar fusion requiring second operation.
Whereas degeneration of the segment adjacent to lumbar fusion has been often seen on radiographs, a small number of patients with such degenerative changes undergo reoperation. Most follow-up studies have focused on adjacent segment disease based on analysis of radiographs. The present study was conducted to understand the pathology of reoperation cases of adjacent segment disease and factors associated with this condition. Operative indication was consistently restricted to patients with neurological involvement. The subjects were 117 patients who had undergone posterior lumbar fusion and were followed for a minimum of 2 years (mean 7 years). Among them, nine patients (7.7%) required a second operation owing to symptomatic adjacent segment disease (stenosis). The reoperation rate was assessed in relation to sex, age, initial pathologic condition, and initial spinal fusion and decompression methods. Data were analyzed in a 2 x 2 cross contingency table using Fisher's exact probability test. A probability of <0.05 was defined as statistically significant. Of the variables examined, only multilevel fusion was associated with a high rate of reoperation with statistical significance (P < 0.04). Two patients (100%) suffering from loss of coronal balance (degenerative scoliosis) also required a second operation. The reoperation rate of 7.7% for adjacent segment disease in this study was consistent with the prevalence of adjacent segment stenosis in the literature. Given the risk of later occurrence of adjacent segment stenosis following multisegment posterolateral fusion, correction of coronal and sagittal balance, preventive decompression of the adjacent segment, or selective decompression without fusion may have to be considered as an additional or alternative procedure.


Herpes simplex virus encephalitis. Diagnostic and treatment considerations.
The patient with meningoencephalitis should be evaluated carefully for the presence of focal signs referable to involvement of the frontotemporal regions of the brain. A significant percentage of cases of encephalitis with discrete focal features are caused by infection with herpes simplex virus. If focal signs are not present, the patient should be managed conservatively but examined regularly because focal neurologic dysfunction may present at any time in the course of disease. If there is clinical and electrographic evidence of involvement of the frontotemporal lobes, radiographic imaging is indicated as well as studies of the serum and cerebrospinal fluid for antibodies indicative of a recent infection with herpes simplex virus. Assuming that the radiographic scans identify the characteristic changes of a focal encephalitis and that the antibody responses are indicative of a recent herpes simplex virus infection, brain biopsy should be done in order to confirm the diagnosis. With positive evidence for HSE, treatment with ara-A should be initiated and continued for 10 days. If the biopsy proves negative for virus, ara-A should be discontinued and the patient managed conservatively (Fig. 3). Although the NIAID study of ara-A treatment of HSE is encouraging, the numbers are small and the evidence is, at best, only suggestive. It is reasonable to use this drug until a better one becomes available for the treatment of known HSE, and treatment should be instituted early before cellular injury is extensive. In centers familiar with this problem, biopsy confirmation of the diagnosis is a simple and informative procedure and can be defended. If patients cannot be moved to a center of this type, physicians familiar with the many facets of this problem should be consulted, and a decision regarding biopsy and treatment should be individualized in light of the circumstances. Biopsy should only be undertaken when the procedure can be done with minimal risk to the patient and the assurance that the maximum amount of information can be gained.


Assessment of Husbandry Practices That Can Reduce the Negative Effects of Exposure to Low Ammonia Concentrations in Broiler Houses.
Ammonia is an important pollutant emitted by broiler litter that can accumulate inside farms, impairing animal health and welfare productivity. An experiment was designed to evaluate of precision husbandry practices such as the effect of ventilation, animal density and growth rate as management options to reduce the adverse effects of ammonia exposure on productive parameters in broiler houses. Two identical experimental rooms were used in this study. They were programmed to differ in ammonia concentration from day 32 of the growing period (10 and 20 ppm in Room 1 and Room 2, respectively). Three treatments were tested in each room: slow growth in high stocking density (SHD), fast growth in low density (FLD) and fast growth in high density (FHD). Animal weight, feed intake and feed conversion ratio were determined weekly. In addition, the immune status of animals was assessed by weighing the organs related to immune response as stress indicators. Increasing ventilation was effective to control ammonia concentrations. Exposure to ammonia caused no significant effect on productive parameters. However, lowering stocking density improved response to higher ammonia concentrations by lowering the feed conversion ratio. No other relevant effects of differential exposure to ammonia were found in fast-growing animals, either at high or low stocking density. The use of slow-growing breeds had no effect on production parameters. Despite having a slower growth rate, their feed conversion ratio was not different from that of fast-growing breeds. The productive performance of slow-growing animals was not affected by the differential exposure to ammonia, but the reduced spleen size would suggest an impairment of the immune system.


On the use of ultrasonic fatigue testing technique--variable amplitude loadings and crack growth monitoring.
In the very high cycle fatigue regime high-frequency testing techniques are required. Using the ultrasonic fatigue technique, testing time could be reduced significantly in comparison to conventional servo-hydraulic machines. An ultrasonic fatigue testing system developed by the BOKU Vienna with load frequencies of about 20kHz is used for variable amplitude loading investigations in the VHCF regime. Therefore, the amplitude level during fatigue tests is controlled by a PC using an own developed software. Additionally, an in situ reconstruction of a damage equivalent load spectrum based on a load time history is introduced schematically. To optimize the experimental procedure a temperature-controlled pulse-pause adaption has been developed and implemented into the software. For quantifying the influence of variable amplitude loadings on the fatigue life, e.g. load interaction effects, the crack growth is measured by using the potential drop technique that is adapted to the ultrasonic fatigue testing system. Finally, the results of two-step-block loading tests are presented.