How do voltage-gated sodium channels enhance migration and invasiveness in cancer cells?
Voltage-gated sodium channels are abnormally expressed in tumors, often as neonatal isoforms, while they are not expressed, or only at a low level, in the matching normal tissue. The level of their expression and their activity is related to the aggressiveness of the disease and to the formation of metastases. A vast knowledge on the regulation of their expression and functioning has been accumulated in normal excitable cells. This helped understand their regulation in cancer cells. However, how voltage-gated sodium channels impose a pro-metastatic behavior to cancer cells is much less documented. This aspect will be addressed in the review. This article is part of a Special Issue entitled: Membrane channels and transporters in cancers.


A survey of the current practice of the informed consent process in general surgery in the Netherlands.
Additional non-English language abstract (in Dutch) A properly conducted surgical informed consent process (SIC) allows patients to authorize an invasive procedure with full comprehension of relevant information including involved risks. Current practice of SIC may differ from the ideal situation. The aim of this study is to evaluate whether SIC practiced by Dutch general surgeons and residents is adequate with involvement of all required elements. All members of the Dutch Society of Surgery received an online multiple choice questionnaire evaluating various aspects of SIC. A total of 453 questionnaires obtained from surgeons and residents representing >95% of all Dutch hospitals were eligible for analysis (response rate 30%). Knowledge on SIC was limited as only 55% was familiar with all three basic elements ('assessment of preconditions', 'provision of information' and 'stage of consent'). Residents performance was inferior compared to surgeons regarding most aspects of daily practice of SIC. One in 6 surgeons (17%) had faced a SIC-related complaint in the previous five years possibly illustrating suboptimal SIC implementation in daily surgical practice. The quality of the current SIC process is far from optimal in the Netherlands. Surgical residents require training aimed at improving awareness and skills. The SIC process is ideally supported using modern tools including web-based interactive programs. Improvement of the SIC process may enhance patient satisfaction and may possibly reduce the number of complaints. Het doel van het preoperatieve informed consent proces (surgical informed consent, SIC) is om patiënten een weloverwogen en welgeïnformeerde keuze te laten maken over hun operatieve ingreep. De hedendaagse praktijk betreffende SIC staat mogelijk ver van de ideale situatie af. Doel van deze studie is om de dagelijkse praktijk van chirurgen en chirurgen in opleiding betreffende SIC te evalueren en te zien of deze voldoet aan de daarvoor gestelde eisen. METHODE: Alle chirurgen en andere leden van de Nederlandse Vereniging voor Heelkunde ontvingen een online multiple-choice vragenlijst betreffende de belangrijkste aspecten van SIC. RESULTATEN: In totaal waren er 453 bruikbare reacties uit meer dan 95% van alle Nederlandse ziekenhuizen (respons 30%). De kennis over SIC blijkt zeer beperkt. Slechts 55% van de chirurgen bleek bekend met de drie basiselementen van SIC ('beoordelen van de competentie van een patiënt', 'verstrekken van informatie' and 'adequaat vastleggen van de toestemming van de patiënt'). De dagelijkse praktijk liet behoorlijke verschillen tussen alle respondenten zien, maar chirurgen in opleiding scoorden significant slechter vergeleken met chirurgen. 17% van alle chirurgen kreeg de afgelopen vijf jaar te maken met een klacht betreffende SIC, wat zou kunnen wijzen op een suboptimale implementatie van SIC in de dagelijkse praktijk. De kwaliteit van het preoperatieve informed consent proces is in Nederland verre van goed. Chirurgen in opleiding scoorden minder goed dan chirurgen en dienen beter geschoold te worden. Het hele SIC proces zou geformaliseerd moeten worden in protocollen. Moderne hulpmiddelen zoals interactieve softwareprogramma's kunnen hierbij mogelijk helpen. Door het verbeteren van het SIC proces kan de patiëntentevredenheid verhoogd worden terwijl het aantal klachten mogelijk wordt verminderd.


Droplet microfluidics on analysis of pathogenic microbes for wastewater-based epidemiology.
Infectious diseases caused by pathogenic microbes have posed a major health issue for the public, such as the ongoing COVID-19 global pandemic. In recent years, wastewater-based epidemiology (WBE) is emerging as an effective and unbiased method for monitoring public health. Despite its increasing importance, the advancement of WBE requires more competent and streamlined analytical platforms. Herein we discuss the interactions between WBE and droplet microfluidics, focusing on the analysis of pathogens in droplets, which is hard to be tackled by traditional analytical tools. We highlight research works from three aspects, namely, quantitation of pathogen biomarkers in droplets, single-cell analysis in droplets, and living cell biosensors in droplets, as well as providing future perspectives on the synergy between WBE and droplet microfluidics.


The rate of early intrauterine growth in twin gestation.
With the use of specimens obtained by induced abortion, the relationships between crown-rump length on the one hand and body weight and menstrual age on the other were investigated in 43 twin fetuses with menstrual ages between 8 1/2 and 21 weeks. This study indicates that the body weight of twins in comparison with length is the same as that of singletons, an indication of the fact that they are not growth retarded. On the other hand, the average crown-rump length when compared with menstrual age is somewhat less than that of singleton fetuses. The possible implications of these findings are discussed in the light of prevailing concepts concerning the growth of twins.


Optimal Sizing of Aortic Stent Graft for Blunt Thoracic Aortic Injury Considering Hypotension-Related Decrease in Aortic Diameter.
To evaluate the optimal sizing of an aortic stent graft in patients with blunt thoracic aortic injury (BTAI), considering the decrease in diameter in hypovolemic status. From 2014 to 2020, 25 patients who underwent thoracic endovascular aortic repair (TEVAR) for BTAI were included. Hemodynamic parameters in the emergency room (ER) and just before the main procedure (MP) were collected. The aortic sizes were measured during initial computed tomography (CT) on arrival in the ER, aortography (AG) during TEVAR, and final CT in the outpatient clinic. The appropriateness of the inserted stent graft size was investigated. The mean values of the final CT/initial CT and final CT/initial AG (proximal descending thoracic aorta [pDTA]) were 113% and 105%, respectively. The final CT/initial CT (pDTA; 122.2% vs 108.8%, p=0.01) and final CT/initial AG (pDTA; 113.4% vs 102.1%, p<0.01) were significantly higher in patients with systolic blood pressure (SBP; MP) ≤90 mm Hg. The final CT/initial CT (pDTA; 120.4% vs 109.0%, p=0.03) and final CT/initial AG (pDTA; 111.4% vs 102.6%, p=0.01) were significantly higher in patients with mean blood pressure (MBP; MP) ≤70 mm Hg. On an average, the inserted stent grafts were oversized by 130% on initial AG. Based on the final CT scan, the inserted stent graft was as large as 122%. In the case of hemodynamic instability with SBP (MP) ≤90 mm Hg or MBP (MP) ≤70 mm Hg, despite adequate resuscitation, an oversized TEVAR stent graft of 130% can reduce the occurrence of endoleak and is sufficiently safe. Despite sufficient resuscitation, the aorta size measured during TEVAR in patients with hemodynamic instability with systolic BP <90 mmHg and mean BP <70 mmHg may be reduced by more than 15% compared to that in the normal state. In this study, the mean size of the stent grafts were oversized by 130% on initial aortography, but were oversized by 122% based on final CT. When the stent graft was oversized by 130% in TEVAR for hemodynamic unstable patient with BTAI, the patient reached the proper oversizing subsequent to hemodynamic recovery.


Persistence of new insecticides and their efficacy against insect pests of okra.
Persistence and efficacy of bifenthrin (25 and 50 g ai ha(-1)), fipronil (50 and 100 g ai ha(-1)) and indoxacarb (70 and 140 g ai ha(-1)) has been studied in okra fruits. The initial deposits varied from 0.259-0.382 microg g(-1) at low and 0.461-0.688 microg g(-1) at high rate of application. The residues persisted upto 10 days with half-life of 1.32-1.58 days for bifenthrin, 0.65-1.12 days for fipronil and 0.58-1.02 days for indoxacarb. Based on ADI, the suggested waiting period was 1 day for bifenthrin and indoxacarb and 3 days for fipronil. All the insecticides were found effective against leafhopper and shoot and fruit borer.


An experimental study on cupular function: mapping of the cupula by direct stimulation.
In the present study, the posterior semicircular canal of the frog was isolated and the entire cupula was exposed after cutting the ampullary wall. The surfaces of the utricle and the lateral cupula were depressed by a fine glass pipette. Action potentials were taken from the posterior ampullary nerve and were compared by using nine points of stimulation on the cupular surface of the utricular side and two points on the lateral surface. The maximum action potential was recorded at stimulus to the lowest point on the utricular side surface. This potential proportionally decreased as the stimulation point approached the top of the cupula. Depression of the lateral surface evoked only a small potential. In contrast to the stimulus applied to the utricular surface, a stimulus to the upper point of the lateral surface yielded a greater potential than did a stimulus to the lower point.


Medical care classification systems in the ambulatory care environment: an evaluative framework.
An essential need of a Quality Assurance Program (QAP) in an ambulatory care setting is accurate and reliable information characterizing the encounter between patient and provider. This information includes identification of the patient's reason for visit, the provider's diagnostic impressions, and procedures performed. Such data can be used to maintain an informational index on the nature of patient care. Based upon this index, a wide variety of special studies in the assessment and assurance of the quality of care can be conducted. A primary consideration in the development of such an index is the choice of a Medical Care Classification System (MCCS) to be used to code encounter-related data. The objective of this paper is to provide a methodology and evaluative framework in which available Medical Care Classification Systems can be assessed as to which one best meets the needs of the quality assurance programs within ambulatory care settings.


Myrrhone inhibits the progression of hepatic fibrosis by regulating the abnormal activation of hepatic stellate cells.
We focus on exploring the antihepatic fibrosis effect of Myrrhone (Myr), a compound extracted from myrrh, and its effective target. Mouse hepatic stellate cells (HSCs) were cultured in vitro and activated by transforming growth factor-β induction. After Myr intervention, cell viability was assessed by the Cell Counting Kit-8 assay. The α-smooth muscle actin(α-SMA) and Collagen I levels were measured by immunofluorescence, and the expressions of tumor necrosis factor-α, interleukin-6, and matrix metalloproteinase-9 were examined by enzyme-linked immunosorbent assay, and the p-Smad3 protein level in HSCs was determined by Western Blot. Small molecule-protein docking and pull-down experiments were conducted to validate the binding capacity between Nard and Smad3. In animal experiments, a mouse model of hepatic fibrosis was established with carbon tetrachloride. Myr was administered by gavage daily to determine the serum alanine aminotransferase and aspartate transaminase levels. The severity of hepatic fibrosis was evaluated by Masson staining, the α-SMA and Collagen I expressions were measured by immunohistochemistry, and the histopathological changes were examined by Sirius red and hematoxylin and eosin staining. Myr suppressed the abnormal activation of HSCs, inhibited the cell viability, downregulated the α-SMA and Collagen I, and inhibited the p-Smad3 expression. After silencing Smad3, the effect of Myr was inhibited. Molecular docking and pull-down experiments revealed the presence of a targeted binding relationship between Myr and Smad3. In mouse experiments, Myr could inhibit hepatic fibrosis. This study discovers that Myr can affect the phosphorylation of Smad3, and inhibit the activation of HSCs and the progression of hepatic fibrosis.


Swimming performance of Cyprinus carpio (Carp) in China.
Cyprinus carpio (Carp) is a fish of great economic importance in China. However, its population has declined considerably due to the construction of barrages. Thus, fishways need to be constructed at barrages to protect fish resources. It is essential for the fishway design to study the swimming performance of carp. By applying incremental flow velocities in a glass open-type flume, three indicators of swimming performance of the carp in China with the body length (BL) of 13-21 cm, including the induced flow velocity (IFV), the critical swimming speed (Ucrit) and the burst swimming speed (Uburst), are systematically assessed. The correlation between the swimming performance and the BL is also analyzed. The results indicate that the IFV of the carp is 15.56 ± 1.79 cm/s, which is not significantly influenced by the BL. The value of Ucrit varies from 60 to 82 cm/s and gradually increases with the increasing value of BL. The relative critical swimming speed (U'crit) is 4.23 ± 0.28 BL/s and gradually decreases with the increasing value of BL. The value of Uburst ranges from 77.2 to 105.1 cm/s, which is linearly positively correlated to BL. The relative burst swimming speed (U'burst) is 5.42 ± 0.39 BL/s. The value of Uburst is approximately 1.28 times of that of Ucrit for the carps with the same BL. These findings are meaningful to the further study of ecological behavior and to the fishway design and optimization of carps.


Nutritional management of cirrhosis patients: A qualitative study exploring perceptions of patients and health workers in Ghana.
Malnutrition is common among patients with end stage liver disease including liver cirrhosis and liver cancer. Optimal nutrition is important to reduce morbidity and mortality of these patients. There is limited qualitative data on nutritional status and management of chronic liver disease patients. We aimed to explore the knowledge, opinions and practices of cirrhosis patients and health workers in nutritional management of cirrhosis in Ghana, in order to determine whether there is a need to improve nutritional care for cirrhosis patients. We conducted a qualitative study using semi-structured interviews of cirrhotic patients (n = 16) and healthcare providers (n = 27) in three academic centers in Accra, Kumasi and Cape Coast (Ghana). Recruitment was by purposive sampling of patients attending specialist liver disease clinics. The recorded data were analyzed using NVivo 11 software, with generation of codes, themes and subthemes. The major themes that emerged from the data included nutrition as part of care delivery during the hospital visit, nutritional recommendations, dietary changes and long-term practice improvement. The results showed that patients and health workers felt dietary recommendations for patients were frequently addressed, but could be significantly improved. We found that in the opinion of study participants, local guidelines are important and necessary in nutritional management of cirrhosis patients, and that participants felt it was difficult to change dietary habits following cirrhosis diagnosis. These results suggest that nutritional management of cirrhosis patients in Ghana requires improvement. Strategies to improve this could include a multi-disciplinary approach to nutritional management, development of local guidelines and continued nutritional assessment, monitoring and follow-up.


[Tracheal lipoma: An exceptional cause of tracheal tumors].
Tracheal tumors are rare, they are most often malignant and can manifest themselves by a non-specific respiratory symptomatology with progressively increasing dyspnea orienting in the first place towards a COPD or even an asthma. Among them, tracheal lipoma is exceptional. Its management is based on removal by rigid bronchoscopy. We report the case of a 73-year-old male patient who presented with non-specific dyspnea that progressively worsened over several months. The EFR showed a flattening of the flow-volume curves, the CT scan showed an anterolateral oval tracheal tumor with fatty density, the bronchial endoscopy showed a tumor lesion with stenosis of about 90% of the airway. Management consisted of a rigid bronchoscopy to delete obtruction with biopsies. Anatomopathology concluded to a tracheal lipoma. Progressively worsening dyspnea, especially if there are signs of inspiratory dyspnea, required a systematic bronchial endoscopy to avoid the possibility of a tracheal tumor.


In vitro development of palatal tissues from embryonic mice. II. Tissue isolation and recombination studies.
The epithelial and mesenchymal tissues of the secondary palate from 12-, 13-, and 14-day embryonic mice were enzymatically separated and cultured in isolation and in homochronic and heterochronic recombinations. In both homochronic and heterochronic recombinations, epithelial differentiation was similar to that in vivo. In heterochronic recombinations, epithelium differentiated according to a schedule appropriate for the age of the epithelium rather than for the age of the mesenchyme, suggesting that differentiation of palatal epithelium is temporally 'determined' as early as 12 days of gestation. Palatal epithelium cultured in isolation was capable of limited differentiation. Potential for differentiation therefore exists within the isolated palatal epithelium at an early stage of palatal development, and epithelial-mesenchymal interactions are required during palatal development to support full epithelial differentiation.


Cyclooxygenase-2 inhibitors induce anoikis in osteosarcoma via PI3K/Akt pathway.
COX-2, an inducible enzyme, is associated with inflammatory diseases and carcinogenesis. Overexpression of COX-2 occurs in many human malignancies, including osteosarcoma. In our study, we reported that Celecoxib, a cyclooxygenase-2 inhibitor, induces apoptosis in human osteosarcoma cell line MG-63 via down-regulation of PI3K/Akt. PI3K/Akt plays an essential role in the cell/extracellar matrix (ECM) and cell/cell adhesion. We hypothesize that COX-2 inhibitors induce anoikis in osteosarcoma via PI3K/Akt, resulted in lack of correct attachment and the down-regulations of β-catenin, TrkB and E-cadherin, which play an essential role in the cell/extracellar matrix (ECM) and cell/cell adhesion. Meanwhile, apoptosis also be disclosed, such as DNA fragments and apoptotic bodies, activation of caspase-8, 9 and cleavage of PARP. With wortmannin, a specific PI3K inhibitor can simulate the effect of COX-2 inhibitors. If our hypothesis is correct, COX-2 inhibitors could cut down the occurrence of metastasis and facilitate the patient who may benefit from addition of COX-2 inhibitors to standard cytotoxic therapy.


Slow-infusion of calcium channel blockers in the emergency management of supraventricular tachycardia.
To compare the efficacy of verapamil and diltiazem as slow infusions in terminating spontaneous supraventricular tachycardia (SVT) in the emergency department (ED). Patients of at least 10 years of age who presented to our ED with regular narrow complex tachycardia not converted with a vagal manoeuvre with an ECG diagnosis of SVT were included. Those who were haemodynamically unstable were excluded. Patients were randomized to undergo either verapamil infusion at a rate of 1 mg/min to a maximum of 20 mg or diltiazem infusion at a rate of 2.5 mg/min to a maximum of 50 mg. Eighty-one patients were randomized to receive verapamil infusion and 80 were randomized to receive the diltiazem infusion. There is no difference in success rate between verapamil (98.8%) and diltiazem (96.3%) infusion. The dose of medication required to convert 25,50 and 75% of SVTs were 4.0,5.0 and 8.0 mg for the verapamil infusion and 10.0,12.5 and 17.5 mg for the diltiazem infusion. There was one complication in each group. Calcium channel blockers infusions were safe and efficacious in terminating spontaneous SVT. There was no difference between the success rates of verapamil and diltiazem infusions.


Calibrated liposomal release of the anti-mitotic agent BI-2536 increases the targeting of mitotic tumor cells.
Cancer drugs which are specifically targeted at mitosis have generally under-delivered as a class. One likely reason is that only a small percentage of cancer cells in a tumor are actually dividing at any moment. If this is the case, then prolonged bioavailability in the tumor should significantly increase the efficacy of antimitotic agents. Here, we show that if the Plk1 inhibitor BI 2536 is co-encapsulated in a liposome with a pair of anions, its release rate is dependent on both the identity and stoichiometry of the anions. We created a library of liposomes with varying release rates using this approach and found that liposomal drug release rates correlated inversely with in vitro cancer cell killing. Xenografted mice treated with a single dose of slow-releasing liposomal BI 2536 experienced tumor volume decreases lasting 12 days and complete responses in 20% of mice. Treatment with two doses a week apart increased the response rate to 75%. This approach, which we termed Paired Anion Calibrated Release (PACeR), has the potential to revive the clinical utility of antimitotic cancer drugs which have failed clinical trials.


The repair of DNA single strand breaks in human cells with genetical disorders.
The aim of this work was to make a preliminary examination of some hereditary diseases, related to precocious aging in order to determine whether the defects involved resulted from an impairment of the DNA repair capacity. To test for an impairment in DNA single strand capacity, in cells from patients with different diseases, we have followed the rate of the DNA after X-irradiation by sedimentation on alkaline sucrose gradients. It was found that all cells observed from Lesh Nyhan, cystic fibrosis, trisomy 15 et 21, retinoblastoma, were able to repair the single strand breaks in their DNA to the same extent and at the same initial rate.


l-Histidinium p-toluene-sulfonate.
In the title salt, C6H10N3O2 (+)·C7H7O3S(-), the imidazole ring makes a dihedral angle of 70.93 (12)° with the plane of the toluene ring. In the crystal, the ions are linked via N-H⋯O and weak C-H⋯O hydrogen bonds forming two-dimensional networks lying parallel to (001). These networks are linked via C-H⋯π inter-actions, forming a three-dimensional structure.


The psychological impact of testing for thrombophilia: a systematic review.
Nowadays, large numbers of patients are tested for thrombophilia, even though the benefits of this strategy remain unclear. A potential disadvantage of this predominantly genetic testing is the psychological impact, including fear, depression and worry. To systematically review studies that determined the nature and extent of the psychological impact of testing for thrombophilia. We searched the MEDLINE data base (1966-2008), the EMBASE data base (1985-2008) and the PsychInfo data base (1806-2008) for relevant trials, without language restrictions. Bibliographies of relevant articles were scanned for additional articles. We reviewed all relevant studies that focused on the psychological impact of testing for thrombophilia. Only full papers of studies that included 15 patients or more were considered eligible for this review. Two reviewers independently extracted data and assessed quality. Six studies fulfilled the eligibility criteria. As these studies varied appreciably in methodology, the pooling of data was not possible. Studies of psychological impact of genetic testing for thrombophilia report few negative results, although most assessments were limited to short-term follow-up, or lacked methodological accuracy. No valid conclusions can be drawn about the psychological impact of genetic testing in patients based on the current available literature. Given the large number of patients that are being exposed to testing for thrombophilia, and the uncertain benefits, there is an urgent need for more uniformity in the measurement of the psychological impact of thrombophilia testing.


Usefulness of three-dimensional electron beam computed tomography for evaluating tracheobronchial anomalies in children with congenital heart disease.
This study was undertaken to delineate tracheobronchial anomalies associated with congenital heart disease. From June 1995 to December 2000, 1,245 children with congenital heart disease underwent cardiac electron beam computed tomography with 3-dimensional reconstruction on an independent workstation. Tracheobronchial anomalies are strongly associated with congenital heart disease and accompanying tracheal stenosis is not uncommon. With 3-dimensional reconstruction, electron beam computed tomography provided excellent anatomic definition of the central tracheobronchial abnormalities.


Modulation of thecal progesterone secretion by estradiol-17 beta.
The effects of estradiol-17 beta on progesterone secretion by isolated bovine theca interna were studied. Preparations of theca interna were obtained from preovulatory follicles of Holstein heifers (n = 5) prior to the endogenous LH surge and cultured for 3 days in a defined medium containing various concentrations of estradiol (0, 0.001, 0.01, 0.1, 1, or 10 microgram/ml). Secretion of progesterone by control theca was low during the first 24 h of culture and increased steadily during the subsequent 48 h. The lowest concentration of estradiol (0.001 microgram/ml) significantly stimulated thecal progesterone production, while higher concentrations (0.1, 1, and 10 microgram/ml) significantly inhibited secretion of progesterone. Estradiol content of the follicular fluid of these preovulatory follicles and several others (n = 13) averaged 1.5 microgram/ml +/- 0.1 (SEM). It is concluded that bovine theca interna is capable of secreting progesterone, that high intrafollicular concentrations of estradiol may inhibit thecal progesterone production in vivo prior to the LH surge, and that estradiol concentration may be be an important modulating factor in the control of follicular progesterone secretion.


Establishing models of corticosteroid patterns during the life history of killer whales (Orcinus orca) under human care.
The use of corticosteroids and their metabolites as a physiologic measure of stress in wildlife species is increasing in both in and ex situ populations. However, factors such as season, circadian rhythm, sex and age are also known to influence corticosteroid production in wildlife. Our objective was to evaluate the influence of these variables on serum cortisol, corticosterone, aldosterone and their fecal metabolites in zoo-based killer whales (Orcinus orca). For evaluation of season, sex and age, we examined 30 animals (21 females, 9 males), and for circadian rhythm, we studied 18 animals (10 females, 8 males). Season did not influence corticosteroids (P ≥ 0.19). Circadian rhythm influenced all corticosteroids (P ≤ 0.012). Serum cortisol and corticosterone were highest in the morning and lower by mid-day and evening. Serum aldosterone was lowest during mid-day and highest in the evening. Excreted corticosteroid metabolites were higher in the morning and mid-day compared to evening (P ≤ 0.002). Serum cortisol was higher in males than females (P = 0.011). The inverse was observed for fecal corticosterone metabolites (P = 0.03). Serum corticosterone increased with age in all animals (P = 0.002), but serum cortisol increased with age in males only (P < 0.001). Finally, combining previously published data and these data reported herein, we developed models of corticosteroid patterns for zoo-based killer whales during life history events, including circadian variation, pregnancy and acute stress that may serve as a guide for evaluating stress physiology and animal welfare in this species.


A Possible Novel Effect for Dapagliflozin in the Management of Subcutaneous Insulin Resistance Syndrome: A Report of Two Cases.
Subcutaneous insulin resistance syndrome (SIRS) is a rare condition in which patients poorly respond to subcutaneous (SC) insulin but maintain a normal response to intravenous (IV) insulin. The underlying pathophysiology remains elusive. Several treatment regimens have been tested for the management of SIRS, none of which included a sodium-glucose cotransporter-2 inhibitor (SGLT-2). Two cases of type 1 diabetes initially achieved adequate glycemic control with subcutaneous insulin. Both cases later progressed into recurrent diabetic ketoacidosis that would resolve following IV insulin administration. Further investigation revealed unresponsiveness to SC, but not IV, insulin and the clinical diagnosis of SIRS was established accordingly. HbA1c values for cases 1 and 2 were 11% on 400 units/day of SC insulin, and 12% on 350 - 400 units/day of SC insulin, respectively. The patients required very high doses of intramuscular (IM) insulin. Subsequently, dapagliflozin as adjunct therapy significantly reduced the patients' IM insulin requirements beyond the anticipated dose reduction. Ultimately, case 1 achieved an HbA1c of 7 - 8% on 90 units/day of IM insulin and 10 mg/day of dapagliflozin, and case 2 achieved an HbA1c of 7 - 8% on 120 units/day of IM insulin and 10 mg/day of dapagliflozin. These are the first reported cases of SIRS in which dapagliflozin, an SGLT-2 inhibitor, was used. The substantial reduction in the IM insulin dose following the addition of dapagliflozin in our reported cases of SIRS suggests a possible novel mechanism for dapagliflozin beyond its glucosuric effects. In this report, we present a hypothetical basis for this possible novel mechanism.


The bioadhesion and effects of microplastics and natural particles on growth, cell viability, physiology, and elemental content of an aquatic macrophyte Elodea canadensis.
Microplastics in the aquatic environment can interact with aquatic plants, but the consequences of these interactions are poorly understood. Therefore, the aim of this study was to investigate the effects of microplastics commonly found in the environment, namely polyethylene (PE) fragments, polyacrylonitrile (PAN) fibres, tire wear (TW) particles under a relevant environmental concentration (5000 particles/L) on the growth, cell viability, physiology, and elemental content of the aquatic macrophyte Elodea canadensis. The effects of microplastics were compared to those of natural wood particles. The results showed that all types of microplastics adhered to plant tissues, but the effect on leaves (leaf damage area) was greatest at PE > PAN > TW, while the effect of natural particles was comparable to that of the control. None of the microplastics studied affected plant growth, lipid, carbohydrate, or protein content. Electron transport system activity was significantly higher in plants exposed to PAN fibres and PE fragments, but also when exposed to natural particles, while chlorophyll a content was negatively affected only by PE fragments and TW particles. Elemental analysis of plant tissue showed that in some cases PAN fibres and TW particles caused increased metal content. The results of this study indicated that aquatic macrophytes may respond differently to exposure to microplastics than to natural particles, likely through the combined effects of mechanical damage and chemical stress.


Self-report of domestic violence and forced sex are related to sexual risk behaviors in a sample of juvenile detainees.
Justice-involved youth have higher rates of sexually transmitted infections (STIs), and a higher prevalence of the associated sexual risk behaviors. Sexual risk behaviors are also associated with alcohol and drug use. Research suggests that a history of trauma is an important predictor of alcohol and drug use in youth offenders, and therefore is a likely contributor to sexual risk behavior in this population. The objective of this analysis is to determine the association of trauma, specifically, domestic violence and forced sex, to six sexual risk behaviors and a history of chlamydia among detained youth. The analysis uses data from a convenience sample of detainees assenting to HIV testing conducted December 2016 - August 2017 using the state-certified Voluntary Counseling Testing and Referral (VCTR) process. Of the 379 youth that received VCTR at the facility, 308 (81.3%) were used in this analysis. Report of domestic violence was significantly associated with sex under the influence of alcohol and was also significantly associated with sex under the influence of marijuana. Forced sex was associated with a sexual partner of unknown HIV status. Traumatic experiences were related to sexual risk behaviors in this analysis, and substance use was strongly implicated in the association. Trauma is known to be a catalyst to sexual risk behaviors, substance use, and delinquency in adolescence. Results support the findings of other investigators and re-iterate the need for trauma-informed interventions that can improve the life trajectories of detained youth.


Cori meets Dobzhansky: Evolution and Gene Expression in St. Louis: A report on the "Evolution and Core Processes in Gene Regulation" meeting in St. Louis, June 25-28, 2015.
St. Louis and its famous Gateway Arch were the setting of the Special Symposium: Evolution and Core Processes in Gene Regulation, sponsored by the American Society for Biochemistry and Molecular Biology. Biochemists and evolutionary biologists highlighted growing connections between studies of biochemical mechanism and natural selection on gene expression.


Parallel Inhibition of Dopamine Amacrine Cells and Intrinsically Photosensitive Retinal Ganglion Cells in a Non-Image-Forming Visual Circuit of the Mouse Retina.
An inner retinal microcircuit composed of dopamine (DA)-containing amacrine cells and melanopsin-containing, intrinsically photosensitive retinal ganglion cells (M1 ipRGCs) process information about the duration and intensity of light exposures, mediating light adaptation, circadian entrainment, pupillary reflexes, and other aspects of non-image-forming vision. The neural interaction is reciprocal: M1 ipRGCs excite DA amacrine cells, and these, in turn, feed inhibition back onto M1 ipRGCs. We found that the neuropeptide somatostatin [somatotropin release inhibiting factor (SRIF)] also inhibits the intrinsic light response of M1 ipRGCs and postulated that, to tune the bidirectional interaction of M1 ipRGCs and DA amacrine cells, SRIF amacrine cells would provide inhibitory modulation to both cell types. SRIF amacrine cells, DA amacrine cells, and M1 ipRGCs form numerous contacts. DA amacrine cells and M1 ipRGCs express the SRIF receptor subtypes sst(2A) and sst4 respectively. SRIF modulation of the microcircuit was investigated with targeted patch-clamp recordings of DA amacrine cells in TH-RFP mice and M1 ipRGCs in OPN4-EGFP mice. SRIF increases K(+) currents, decreases Ca(2+) currents, and inhibits spike activity in both cell types, actions reproduced by the selective sst(2A) agonist L-054,264 (N-[(1R)-2-[[[(1S*,3R*)-3-(aminomethyl)cyclohexyl]methyl]amino]-1-(1H-indol-3-ylmethyl)-2-oxoethyl]spiro[1H-indene-1,4'-piperidine]-1'-carboxamide) in DA amacrine cells and the selective sst4 agonist L-803,087 (N(2)-[4-(5,7-difluoro-2-phenyl-1H-indol-3-yl)-1-oxobutyl]-L-arginine methyl ester trifluoroacetate) in M1 ipRGCs. These parallel actions of SRIF may serve to counteract the disinhibition of M1 ipRGCs caused by SRIF inhibition of DA amacrine cells. This allows the actions of SRIF on DA amacrine cells to proceed with adjusting retinal DA levels without destabilizing light responses by M1 ipRGCs, which project to non-image-forming targets in the brain.


Development of Freeze-Drying Cycles for Pharmaceutical Products Using a Micro Freeze-Dryer.
This article aims to investigate how a small-scale freeze-dryer can be used for process design. The system encompasses a temperature controlled metallic ring that surrounds a small batch of vials, in contact with the external vials through removable thermal conductors. The temperature of the ring can be modified to keep a constant difference with the temperature of one or more vials of the batch. In this article, an extensive validation of the system is given, considering 10% w/w sucrose and 5% w/w mannitol solutions, processed in different types of vials (6 R and 20 R) and in different operating conditions. The micro freeze-dryer was also shown to be able to provide accurate estimates of the overall heat transfer coefficient from the shelf to the product in the vials (Kv) and of the resistance of the dried cake to vapor flux (Rp): both values appeared to be very close to those obtained for the same case studies in a pilot-scale unit. Finally, the use of the micro freeze-dryer to control product temperature and drying time values to simulate a pilot-scale unit was addressed, thus demonstrating the adequacy of this system for process scaleup.


Long-term light and moderate exercise intervention similarly prevent both hippocampal and glycemic dysfunction in presymptomatic type 2 diabetic rats.
A prediabetic population has an increased risk of cognitive decline and type 2 diabetes mellitus (T2DM). This study investigated whether the progression of memory dysfunction and dysregulated brain glycogen metabolism is prevented with 4 mo of exercise intervention from the presymptomatic stage in a T2DM rat model. Memory function and biochemical and molecular profiles were assessed in the presymptomatic stage of Otsuka-Long-Evans-Tokushima fatty (OLETF) rats, a T2DM model, with Long-Evans Tokushima (LETO) rats as genetic control. These rats were subjected to light- or moderate-intensity treadmill running for 4 mo with repetition of the same experiments. Significant hippocampal-dependent memory dysfunction was observed in the presymptomatic stage of OLETF rats, accompanied by downregulated levels of hippocampal monocarboxylate transporter 2 (MCT2), a neuronal lactate-transporter, without alteration in hippocampal glycogen levels. Four months of light or moderate exercise from the presymptomatic stage of T2DM normalized glycemic parameters and hippocampal molecular normalization through MCT2, glycogen, and brain-derived neurotrophic factor (BDNF) levels with the improvement of memory dysfunction in OLETF rats. A 4-mo exercise regimen from the presymptomatic stage of T2DM at a light and moderate intensities contributed to the prevention of the development of T2DM and the progression of cognitive decline with hippocampal lactate-transport and BDNF improvement.NEW & NOTEWORTHY Type 2 diabetes mellitus is an independent risk factor for hippocampal memory dysfunction, which would progress since the prediabetic stage. We found that 4 mo of exercise both at the light and moderate intensity prevented the progression of memory dysfunction with an improvement of hippocampal MCT2 expression in presymptomatic diabetes, implying that light intensity exercise could be a therapeutic approach, and the alteration of hippocampal MCT2 would be a therapeutic target of memory dysfunction from presymptomatic diabetes.


Poorly differentiated carcinomas of the thyroid with trabecular, insular, and solid patterns: a clinicopathologic study of 183 patients.
The term poorly differentiated (PD) carcinoma was proposed 20 years ago to define aggressive, follicular-derived thyroid carcinomas with behavior intermediate between follicular/papillary and anaplastic carcinomas. Among the variable histologic patterns recognized in such tumors, trabecular-insular-solid (TIS) areas usually are predominant. Conversely, some authors pointed out that PD carcinomas are characterized by unequivocal, high-grade histology with atypias, high mitotic counts, and necrosis rather than by a specific growth pattern. The clinicopathologic features of a series of 183 thyroid carcinomas with predominant (n = 165 tumors) or focal (n = 18 tumors) TIS growth patterns were studied by univariate and multivariate overall survival analyses and were compared with clinical outcomes. Subgroups included tumors with predominant oxyphilic features (n = 66 tumors) and (residual) papillary carcinoma features (n = 24 tumors). Control groups of papillary (n = 68 tumors), follicular (n = 71 tumors), and anaplastic (n = 35 tumors) carcinomas also were included for overall survival analysis. TIS carcinomas had an intermediate behavior between papillary/follicular and anaplastic carcinomas (P < 0.0001). Univariate and multivariate statistical analyses demonstrated that age > 45 years (P = 0.007), the presence of necrosis (P < 0.0001), and a mitotic count > 3 per 10 high-power fields (P = 0.01) were associated with poor outcome. A simplified scoring system based on statistically significant parameters allowed the identification of three prognostic subgroups (P < 0.0001). PD TIS carcinomas overall followed a more aggressive course compared with differentiated thyroid carcinomas, irrespective of the extent of the TIS component. However, a numeric scoring system applied to specific clinicopathologic parameters further may identify three prognostic categories of patients who have significantly different survival rates at 5 years and 10 years.


Voltage-gated calcium channels in adult rat inferior colliculus neurons.
The inferior colliculus (IC) plays a key role in the processing of auditory information and is thought to be an important site for genesis of wild running seizures that evolve into tonic-clonic seizures. IC neurons are known to have Ca(2+) channels but neither their types nor their pharmacological properties have been as yet characterized. Here, we report on biophysical and pharmacological properties of Ca(2+) channel currents in acutely dissociated neurons of adult rat IC, using electrophysiological and molecular techniques. Ca(2+) channels were activated by depolarizing pulses from a holding potential of -90 mV in 10 mV increments using 5 mM barium (Ba(2+)) as the charge carrier. Both low (T-type, VA) and high (HVA) threshold Ca(2+) channel currents that could be blocked by 50 microM cadmium, were recorded. Pharmacological dissection of HVA currents showed that nifedipine (10 microM, L-type channel blocker), omega-conotoxin GVIA (1 microM, N-type channel blocker), and omega-agatoxin TK (30 nM, P-type channel blocker) partially suppressed the current by 21%, 29% and 22%, respectively. Since at higher concentration (200 nM) omega-agatoxin TK also blocks Q-type channels, the data suggest that Q-type Ca(2+) channels carry approximately 16% of HVA current. The fraction of current (approximately 12%) resistant to the above blockers, which was blocked by 30 microM nickel and inactivated with tau of 15-50 ms, was considered as R-type Ca(2+) channel current. Consistent with the pharmacological evidences, Western blot analysis using selective Ca(2+) channel antibodies showed that IC neurons express Ca(2+) channel alpha(1A), alpha(1B), alpha(1C), alpha(1D), and alpha(1E) subunits. We conclude that IC neurons express functionally all members of HVA Ca(2+) channels, but only a subset of these neurons appear to have developed functional LVA channels.


A family of conserved bacterial virulence factors dampens interferon responses by blocking calcium signaling.
Interferons (IFNs) induce an antimicrobial state, protecting tissues from infection. Many viruses inhibit IFN signaling, but whether bacterial pathogens evade IFN responses remains unclear. Here, we demonstrate that the Shigella OspC family of type-III-secreted effectors blocks IFN signaling independently of its cell death inhibitory activity. Rather, IFN inhibition was mediated by the binding of OspC1 and OspC3 to the Ca2+ sensor calmodulin (CaM), blocking CaM kinase II and downstream JAK/STAT signaling. The growth of Shigella lacking OspC1 and OspC3 was attenuated in epithelial cells and in a murine model of infection. This phenotype was rescued in both models by the depletion of IFN receptors. OspC homologs conserved in additional pathogens not only bound CaM but also inhibited IFN, suggesting a widespread virulence strategy. These findings reveal a conserved but previously undescribed molecular mechanism of IFN inhibition and demonstrate the critical role of Ca2+ and IFN targeting in bacterial pathogenesis.


Cholangiographic findings in deteriorating primary sclerosing cholangitis.
The evolution of primary sclerosing cholangitis (PSC) is difficult to predict, and ERCP may be helpful in diagnosing its complications and treating them. Few descriptions of cholangiographic findings during the course of the disease have yet appeared. We report here on our experience in the management and follow-up by ERCP of six patients with deteriorating PSC. Between 1981 and 1992, PSC was diagnosed in 18 patients. Clinical, biochemical, and cholangiographic findings were recorded during the follow-up period. An initial group of 12 patients had a good clinical and biochemical course (mean 56 months) and did not require systematic control ERCPs. A second group of six patients, showing cholangiographic features similar to those of the first group at presentation, developed clinical and biochemical deterioration within a mean period of 43 months (range 6-96). These six patients had a significant elevation of serum bilirubin and at ERCP showed worsening bile duct lesions, with marked strictures and a subsequent increase in the maximum duct diameter in both the extrahepatic and intrahepatic ducts compared to the initial ERCPs. In two of these six patients, the strictures were due to a superimposed cholangiocarcinoma. Endoscopic drainage was unsuccessful, transplantation impracticable due to extrahepatic spread, and both patients died three and five months, respectively, after diagnosis of cholangiocarcinoma. By contrast, four patients with dominant extrahepatic duct involvement benefited from endoscopic procedures (mean follow-up 19 months) without any recurrence of cholangitis and without requiring long-term antimicrobial treatment. The follow-up checks included regular ERCPs performed every six months for duct and stent assessment, and cytology brushing. Since there are no effective means of early and accurate detection of cholangiocarcinoma, the use of ERCP to compare the course of cholangiographic lesions over time may be of interest. Patients with extrahepatic duct involvement may benefit from endoscopic therapy. By contrast, in patients with dominant intrahepatic duct disease and a deteriorating clinical status, attempts at endoscopic therapy are not helpful, and liver transplantation needs to be considered promptly.


Dynamic stomatal behavior and its role in carbon gain during lightflecks of a gap phase and an understory Piper species acclimated to high and low light.
Steady-state and dynamic stomatal and assimilation responses to light transients were characterized in sun- and shade-acclimated plants of Piper auritum, a pioneer tree, and Piper aequale a shade-tolerant shrub from a tropical forest at Los Tuxtlas, Veracruz, México. Despite essentially identical steady-state responses of stomatal conductance to PFD of P. aequale and P. auritum shade plants, the dynamic responses to lightflecks were markedly different and depended on the growth regime. For both species from both growth environments, the increase in stomatal conductance occurring in response to a lightfleck continued long after the lightfleck itself so that the maximum stomatal conductance was not reached until 20-40 min after the lightfleck. Closing then occurred until stomatal conductance returned to near its original value before the lightfleck. Plants that were grown under light regimes similar to those of their natural habitat (high light for P. auritum and shade for P. aequale) had large maximum excursions of stomatal conductance and slower closing than opening responses. Plants grown under the opposite conditions had smaller excursions of stomatal conductance, especially in P. auritum, and more symmetrical opening and closing. The large and hysteretic response of stomatal conductance of P. aequale shade plants to a lightfleck was shown to improve carbon gain during subsequent lightflecks by 30-200%, depending on lightfleck duration. In contrast the very small stomatal response to lightflecks in P. auritum shade plants, resulted in no significant improvement in use of subsequent lightflecks.


[Risk of electromagnetic fields in control board and switchboard rooms at petrochemical plants].
Authors evaluate electromagnetic field exposure (in the range (5-30,000 Hz) in control board and switchboard rooms of petrochemical plants. According to measured values and international guidelines, they conclude that operators should be exposed without adverse effects.


On the use of genetic algorithms for response surface modeling in high-performance liquid chromatography and their combination with the Microsoft Solver.
Four genetic algorithms--the classical, Haupt's, Brunetti's and a modification of the classical algorithm suggested in the present paper--are examined when they are used for the modeling of response surfaces in high-performance liquid chromatography (HPLC). We found that the best results are obtained from our modification and the worst by Haupt's algorithm. The classical genetic algorithm gives satisfactory results, better than those of Brunetti's algorithm. We also ascertained that all genetic algorithms may get stuck in a local minimum other than the global one, except for our modification, which can be considered to approach a global method. Finally, the time needed for the optimization of a genetic algorithm and the combination of a genetic algorithm with a non-linear least-squares routine are considered and discussed.


Identifying Opioid Use Disorder in the Emergency Department: Multi-System Electronic Health Record-Based Computable Phenotype Derivation and Validation Study.
Deploying accurate computable phenotypes in pragmatic trials requires a trade-off between precise and clinically sensical variable selection. In particular, evaluating the medical encounter to assess a pattern leading to clinically significant impairment or distress indicative of disease is a difficult modeling challenge for the emergency department. This study aimed to derive and validate an electronic health record-based computable phenotype to identify emergency department patients with opioid use disorder using physician chart review as a reference standard. A two-algorithm computable phenotype was developed and evaluated using structured clinical data across 13 emergency departments in two large health care systems. Algorithm 1 combined clinician and billing codes. Algorithm 2 used chief complaint structured data suggestive of opioid use disorder. To evaluate the algorithms in both internal and external validation phases, two emergency medicine physicians, with a third acting as adjudicator, reviewed a pragmatic sample of 231 charts: 125 internal validation (75 positive and 50 negative), 106 external validation (56 positive and 50 negative). Cohen kappa, measuring agreement between reviewers, for the internal and external validation cohorts was 0.95 and 0.93, respectively. In the internal validation phase, Algorithm 1 had a positive predictive value (PPV) of 0.96 (95% CI 0.863-0.995) and a negative predictive value (NPV) of 0.98 (95% CI 0.893-0.999), and Algorithm 2 had a PPV of 0.8 (95% CI 0.593-0.932) and an NPV of 1.0 (one-sided 97.5% CI 0.863-1). In the external validation phase, the phenotype had a PPV of 0.95 (95% CI 0.851-0.989) and an NPV of 0.92 (95% CI 0.807-0.978). This phenotype detected emergency department patients with opioid use disorder with high predictive values and reliability. Its algorithms were transportable across health care systems and have potential value for both clinical and research purposes.


Radical and ionic meta-C-H functionalization of pyridines, quinolines, and isoquinolines.
Carbon-hydrogen (C-H) functionalization of pyridines is a powerful tool for the rapid construction and derivatization of many agrochemicals, pharmaceuticals, and materials. Because of the inherent electronic properties of pyridines, selective meta-C-H functionalization is challenging. Here, we present a protocol for highly regioselective meta-C-H trifluoromethylation, perfluoroalkylation, chlorination, bromination, iodination, nitration, sulfanylation, and selenylation of pyridines through a redox-neutral dearomatization-rearomatization process. The introduced dearomative activation mode provides a diversification platform for meta-selective reactions on pyridines and other azaarenes through radical as well as ionic pathways. The broad scope and high selectivity of these catalyst-free reactions render these processes applicable for late-stage functionalization of drugs.


Muscle damage is linked to cytokine changes following a 160-km race.
Muscle damage and perceived soreness following the 160-km Western States Endurance Run were related to changes in plasma cytokines and use of nonsteroidal anti-inflammatory drugs (NSAIDS). Subjects included 60 ultramarathoners (mean+/-SE, age 45.3 +/- 1.1 years) who finished the race in under 30 h (26.3 +/- 0.4 h). Blood samples were collected the morning prior to and immediately following the race, and subjects recorded muscle soreness during the week following the race using a 10-point Likert scale (DOMS). Seven plasma cytokines were measured including IL-6, IL-10, IL-8, IL-1ra, granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein 1 (MCP-1), and macrophage inflammatory protein 1beta (MIP-1beta). Cytokine changes were compared between NSAID users and nonusers, and correlated with creatine phosphokinase (CPK) and DOMS. Significant increases were measured for all seven cytokines, with the greatest fold increases seen for IL-6 (125x), IL-10 (24x), and G-CSF (12x). CPK was correlated with changes in IL-6, G-CSF, IL-10, IL-1ra, and MCP-1 (r = .49-.68), (P < .001), but not IL-8 or MIP-1beta. DOMS averaged 7.1 +/- 0.3 the day after the race, and 5.0 +/- 0.3, 2.5 +/- 0.2, and 1.6 +/- 0.1 3 days, 5 days, and 7 days post-race, respectively, and each was correlated with CPK (r = .40-.63, P < .001) and changes in IL-6, G-CSF, IL-10, and MCP-1 (r = .28-.77, P < .05). A comparison of NSAID users (72% of athletes) and nonusers showed no differences in CPK or DOMS, but did reveal greater increases in five of seven cytokines in the NSAID users (P < .05). In conclusion, muscle damage in athletes competing in a 160-km race was significantly correlated with post-race DOMS and increases in five of seven cytokines. NSAID users did not experience a reduction in muscle damage or DOMS, but did have higher post-race plasma levels in five of seven cytokines.


Response of bacterial communities to mining activity in the alpine area of the Tianshan Mountain region, China.
Anthropogenic activities, such as mining, influence soil bacterial community composition and microbial distributions. In the current study, the patterns in microbial distribution and the environmental drivers shaping the soil bacterial community composition in the alpine mining area of the Tianshan Mountain region, China, were investigated, and the bacterial communities were analyzed using 16S rDNA pyrosequencing. The environmental factors and their relationships with the microbial community composition, structure, and diversity were also assessed. The soil organic carbon (SOC) concentration increased along the elevation gradient, with the highest concentration in the mining area, which increased microbial abundance and species richness. Some metals, like Ca, Cu, Pb, and Zn, accumulated significantly in the tailing area and were negatively correlated with the microbial community structure. Proteobacteria, Acidobacteria, Actinobacteria, and Verrucomicrobia were the dominant phyla; these dominant phyla were more abundant in the areas without mining than in the areas with mining at the same altitude. The relative abundance of Proteobacteria and Verrucomicrobia significantly increased along the elevation gradient, while that of Actinobacteria in the mining camp area was more than twice those in the other areas due to higher soil pH. Soil biomass was the highest in the valley. Collectively, these results elucidate the influence of anthropogenic mining activities on soil microbial communities in alpine mining soils and provide a basis for the future management of heavy metal-contaminated areas using the indigenous dominant bacterial phyla.


The large late virus transcripts synthesized in herpesvirus suis (pseudorabies) virus-infected cells are not precursors of mRNA.
The sizes of early and late pseudorabies virus transcripts were compared to those of early and late mRNA. While the early primary transcripts were of approximately the same size as early mRNA, a large proportion of late primary transcripts was much larger than late mRNA. Furthermore, most early transcripts were transported efficiently to the cytoplasm and were relatively stable. In contrast, a large proportion of the late transcripts were retained in the nucleus and turned over rapidly. Specific retention in the nucleus of transcripts originating from some regions of the genome could be detected. These were, however, not preferentially degraded; degradation of transcripts originating from all regions of the genome, including those from which late mRNA originates, occurred. Experiments designed to determine whether part of the large transcripts are processed into mRNA revealed that most of the large late transcripts synthesized by the infected cells bear no precursor relationship to mRNA. Thus, during late phase of infection, most regions of the genome are abundantly transcribed as large RNA molecules; these are not destined to be processed into mRNA.


Association of Rare Immune-Related Adverse Events to Survival in Advanced Cancer Patients Treated with Immune Checkpoint Inhibitors: A Real-World Single-Center Cohort Study.
Immune checkpoint inhibitors (ICIs) are associated with immune-related (ir) adverse events (AEs) resembling autoimmune diseases. In this retrospective cohort study of patients (pts) treated with ICIs at Oulu University Hospital from 2014-2020, we analysed the spectrum of severe irAEs and their prognostic nature, focusing on rare irAEs. Pts (n = 173) with lung cancer (n = 76, 43.9%), melanoma (n = 56, 32.4%), renal and bladder cancers (n = 34, 19.7%), head and neck cancers (n = 4, 2.3%), SCC (n = 2, 1.2%), and CRC (n = 1, 0.6%) receiving single anti-PD-(L)1 (n = 160) or combination (ICI-ICI n = 9, ICI-chemotherapy n = 4) therapy were included. The survival analysis focused on single anti-PD-(L)1-treated patients with melanoma, lung cancer, and renal and bladder cancers (n = 142). Grade ≥ 3 irAEs of multiple aetiology occurred in 29 patients treated with single-PD-L1 therapy (20.4%), which was associated with improved progression-free survival (PFS) (HR 0.50, CI 0.31-0.78) but not overall survival (OS) (HR 0.88, CI 0.52-1.50). Rare grade ≥ 3 events occurred in 10 (7.0%) pts with no association with PFS (HR 0.90, CI 0.42-1.94). Hence, the presence of rare grade ≥ 3 irAEs was associated with a tendency for inferior OS (HR 1.44, CI 0.66-3.11). Pts with rare grade ≥ 3 irAEs had inferior OS, possibly reflecting the delay in diagnostic workflow and the treatment of irAEs. One explanation for the high incidence of irAEs could be the Finnish population-based genetic variation affecting the immune system.


An improved universal theoretical formula for intraocular lens power prediction.
Although available empirically derived and theoretical formulas perform adequately for eyes of average axial length, both have been shown to be deficient for eyes that have unusually short and long axial lengths. I developed a formula based on a theoretical model eye in which anterior chamber depth is related to axial length and keratometry. A relationship between the A-constant and a "lens factor" is also used to determine anterior chamber depth. The location of the intraocular lens' principle planes of refraction is retained as a relevant variable in the formula, and the user need not know the material and construction of the lens and or its constant. I compared the new formula with the SRK II, Holladay, and SRK/T formulas in a group of 100 unselected patients and in selected subgroups of patients with average, short, and long axial lengths. The new formula was significantly more accurate than the other third-generation formulas and maintained its accuracy in the subgroups. The formula can be described as universal because it can be used for different lens styles and for eyes with short, medium, and long axial lengths.


True duplicate bladder exstrophy: a case report.
True duplicate exstrophy is a rare and interesting congenital anomaly. We are adding a case to the previously reported seven cases in the world literature.


Radioimmunoassay for salivary carbonic anhydrase in human parotid saliva.
A specific radioimmunoassay for human carbonic anhydrase (CA) VI has been developed and used to determine the concentrations of the enzyme in saliva. The assay detected as little as 200 pg of CA VI and the antibody used did not cross-react with CA II or other salivary proteins. The method showed an intra-assay variation of 8.5% and an inter-assay variation of 16.9%. The concentration in parotid saliva varied over a wide range (from 9.7 micrograms/ml to 121 micrograms/ml) with an average value of 47.0 +/- 39.2 (SD) micrograms/ml (n = 50). The mean secretion rate of CA VI from the combined parotid glands was 42.8 +/- 37.9 micrograms/min. CA VI represented about 3% of the total protein in parotid saliva.


The need for targeted implementation research to improve coverage of basic vaccines and introduction of new vaccines.
The Decade of Vaccines Collaboration (DoVC) Research and Development (R&D) Working Group identified implementation research as an important step toward achieving high vaccine coverage and the uptake of desirable new vaccines. The R&D Working Group noted that implementation research is highly complex and requires participation of stakeholders from diverse backgrounds to ensure effective planning, execution, interpretation, and adoption of research outcomes. Unlike other scientific disciplines, implementation research is highly contextual and depends on social, cultural, geographic, and economic factors to make the findings useful for local, national, and regional applications. This paper presents the broad framework for implementation research in support of immunization and sets out a series of research questions developed through a Delphi process (during a DoVC-supported workshop in Sitges, Spain) and a literature review.


Effect of long-term remote ischaemic conditioning on platelet function and fibrinolysis in patients with chronic ischaemic heart failure.
Remote ischaemic conditioning (RIC) protects against ischaemia-reperfusion injury through cellular protective pathways, but may also modulate haemostasis. We aimed to investigate the effect of long-term RIC on platelet function and fibrinolysis in patients with chronic ischaemic heart failure (CIHF). In a prospective, outcome-assessor blinded, paired study, 16 patients with CIHF and 21 age- and gender-matched controls without ischaemic heart disease (IHD) were treated with RIC once daily for 28±4days. RIC was performed as four cycles of 5min upper arm ischaemia and reperfusion. We evaluated collagen and arachidonic acid induced platelet aggregation (Multiplate® Analyzer), platelet turnover (Sysmex® XE-5000), platelet activation (plasma soluble-platelet-selectin) and fibrinolysis (clot lysis time, tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1)). We compared blood samples assessed at baseline and following long-term RIC. Long-term RIC did not affect platelet aggregation, turnover or activation or PAI-1 in any study groups. Long-term RIC did not affect fibrin clot lysis time in patients with CIHF but reduced fibrin clot lysis time in matched controls without IHD (median: 773s (interquartile range: 689-936) vs. 658s (618-823), p=0.03). t-PA was increased following long-term RIC in CIHF patients (2.5 (1.7-3.4) vs. 2.9 (1.8-4.0), p=0.03) and in matched controls without IHD (1.5 (1.3-1.9) vs. 1.6 (1.4-2.3), p=0.03). While long-term RIC did not affect collagen or arachidonic acid induced platelet aggregation, platelet turnover or sP-selectin, fibrinolysis was increased although most consistently in matched controls without IHD. This finding suggests that RIC may stimulate fibrinolysis potentially reducing the risk of thrombosis.


Recurrent Bilateral Salpingitis in a Sexually Inactive Adolescent: Don't Forget about the Appendix.
Salpingitis is an acute inflammation of the fallopian tubes and is extremely uncommon in patients who are sexually inactive or premenarchal. We describe a 15-year-old sexually inactive patient with recurrent bilateral salpingitis. After the second episode, she underwent an exploratory laparoscopy and was diagnosed with chronic appendicitis. Appendicular involvement should be kept in mind in recurrent salpingitis episodes, especially if previous imaging studies do not show signs of appendicitis. Imaging studies should be repeated in experienced centers in case of recurrent episodes. Laparoscopy is the gold standard for the diagnosis of salpingitis and can confirm appendicular or other abdominal involvement. Early diagnosis can help to prevent recurrent episodes of salpingitis and thus reduce the risk of sequelae.


Differential enhancement of behavioral sensitivity to apomorphine following chronic treatment of rats with (-)-sulpiride and haloperidol.
Rat exploratory activity as well as apomorphine-induced hypermotility and stereotyped behavior were assayed following acute (60 min before) or chronic (21 days) administration of sulpiride stereoisomers and haloperidol. Parallel groups of rats were assayed for their hypermotility and stereotyped responses to challenging doses of apomorphine 7 and 21 days after discontinuation of chronic treatments. Following its acute administration, (-)-sulpiride fully antagonized apomorphine-induced hypermotility without affecting the level of animal spontaneous activity and partially counteracted stereotyped behavior. Haloperidol completely suppressed both apomorphine responses and also depressed exploratory activity. Some tolerance to the anti-apomorphine effect of (-)-sulpiride groups exhibited enhanced behavioral sensitivity to apomorphine only with respect to hypermotility, whereas haloperidol groups were supersensitive with respect to both hypermotility and stereotyped responses. The results are discussed in terms of differential dopamine receptor supersensitivity arising from prolonged administration of butyrophenone and substituted benzamide.


Systematic Targeting of Protein-Protein Interactions.
Over the past decade, protein-protein interactions (PPIs) have gone from being neglected as 'undruggable' to being considered attractive targets for the development of therapeutics. Recent advances in computational analysis, fragment-based screening, and molecular design have revealed promising strategies to address the basic molecular recognition challenge: how to target large protein surfaces with specificity. Several systematic and complementary workflows have been developed to yield successful inhibitors of PPIs. Here we review the major contemporary approaches utilized for the discovery of inhibitors and focus on a structure-based workflow, from the selection of a biological target to design.


Forskolin potentiates the paraoxon-induced hyperexcitability in snail neurons by blocking afterhyperpolarization.
One characteristic of organophosphate poisoning is the ability to increase excitability or induce epileptiform activity in nerve cells, but underlying mechanisms are not fully understood. We have previously reported that paraoxon, an organophosphate compound, at submicromolar concentrations effectively suppress Ca(2+) spikes and modulate the activity of snail neurons. This effect was unrelated to acetylcholinesterase (AChE) inhibition but was found to involve the direct or indirect modulation of ion channels [Vatanparast J, Janahmadi M, Asgari AR, Sepehri H, Haeri-Rohani A. Paraoxon suppresses Ca(2+) spike and afterhyperpolarization in snail neurons: relevance to the hyperexcitability induction. Brain Res 2006a;1083(1):110-7]. In the present study, the interaction of paraoxon with cAMP formation on the modulation of Ca(2+) spikes and neuronal excitability was examined. Forskolin, the activators of adenylate cyclase, suppressed afterhyperpolarization (AHP) and increased the activity of snail neurons without any significant effect on the Ca(2+) spike duration. Pretreatment with forskolin, although attenuated the suppressing effect of paraoxon on the duration of Ca(2+) spikes but also potentiated the paraoxon-induced hyperexcitability by enhancing the suppressive effects of paraoxon on AHP. Our findings support the possible involvement of cAMP formation in the paraoxon-induced AHP suppression and neuronal hyperexcitability, although activation of cAMP pathway may attenuates some effects of paraoxon.


Urine human epidermal growth factor (hEGF) levels following surgery.
In order to investigate the contribution of growth factors in conditions of postoperative stress, we measured the blood and urine hEGF levels in patients with various surgical disorders. While the levels of serum and plasma hEGF did not change significantly after surgery, urine hEGF (u-hEGF) stayed at the base-level followed by a single peak within a postoperative period of two weeks. The peak day of u-hEGF in patients with more than 1000 ml of intraoperative bleeding was later than that in patients with bleeding of less than 1000 ml. In patients with postoperative renal failure or dysfunction, u-hEGF levels dropped and did not rise until the recovery of renal function. The changing pattern of u-hEGF levels was an indicator of operative bleeding volume and renal function.


Antigen specific in vitro activity of lymphocyte subpopulations from Mantoux positive and negative subjects assessed by lymphocyte proliferation assay.
A micromethod for the lymphocyte proliferative assay and multifactorial analysis of stimulation indices classified according to subjects, lymphocyte combinations and antigens was used to elucidate deficient Mantoux (Mx) reactions in BCG-vaccinated subjects. It was possible to group each of the factors so that interactions between the two latter factors were not present within but only between groups. Comparisons between the subject groups showed a clear association between results of this in vitro test with PPD antigen and the Mx test carried out with 1 TU. The in vitro test had the advantage of being more sensitive and of not introducing the immune reactions occasionally seen with the Mx test. Thymus-processed lymphocytes were the cells responsible with functions both as effector and regulator cells, B lymphocytes had only regulatory properties.


Comparison of medical costs generated by IBS patients in primary and secondary care in the Netherlands.
Irritable Bowel Syndrome (IBS) is a functional somatic syndrome characterized by patterns of persistent bodily complaints for which a thorough diagnostic workup does not reveal adequate explanatory structural pathology. Detailed insight into disease-specific health-care costs is critical because it co-determines the societal impact of the disease, enables the assessment of cost-effectiveness of existing and new treatments, and facilitates choices in treatment policy. In the present study the aim was, to compare the costs and magnitude of healthcare consumption for patients diagnosed with Irritable Bowel Syndrome (IBS) in primary and secondary care, compare these costs with the average health care expenditure for patients without IBS and describe these costs in further detail. Reimbursement data for patients diagnosed with IBS by a general practitioner (GP) or specialist between 2006 and 2009 were extracted from a healthcare insurance company and compared to an age and gender matched control group of patients without IBS. Using a case-control design, direct medical costs for GP consultations, specialist care and medication prescriptions were calculated. Data of 326 primary care and 9274 secondary care IBS patients were included in the analysis. For primary care patients, the mean total annual health care costs for the three years after diagnosis compared to the three years before diagnosis, increased with 486 Euro after IBS was diagnosed, whereas for secondary care patients, these costs increased with 2328 Euro. Total health care costs remained higher in the three years after the initial diagnosis when the patient is treated in secondary care, compared to primary care. This increase was significant for hospital specialist costs and medications, but not for GP contacts. For controls, there was no significant difference in mean total annual health costs in the three years before and the three years after the diagnosis and also no significant difference in cost increases between both primary- and secondary-care control patients. Total healthcare costs per patient substantially increase after a diagnosis of IBS and IBS related costs are significantly higher when patients are treated in secondary-care compared to primary-care. IBS patients should be treated in primary-care where possible, not only because guidelines recommend this from a quality of care viewpoint, but also to optimize use of health care resources. Referral should be restricted to those patients with alarm symptoms, with ill-matching symptoms, or other cases of diagnostic uncertainty.


Towards sustainable biodiesel and chemical production: Multifunctional use of heterogeneous catalyst from littered Tectona grandis leaves.
Waste biomass derived heterogeneous catalyst is an excellent alternative to chemically synthesized catalysts. In this work, calcined Tectona grandis leaves were proposed as an eco-friendly, renewable and low cost heterogeneous base catalyst. The prepared catalyst was examined by FTIR, XRD, XPS, SEM, EDX, TEM, TGA, BET and Hammett indicator test. The catalyst has an appealing nature towards various chemical transformations due to its basic surface sites provided by alkali and alkaline earth metals. The efficiency of the catalyst was successfully investigated by its application in biodiesel production. The products were confirmed by 1H and 13C NMR. 100% FAME conversion was attained using a catalyst loading of 2.5 wt% under optimized reaction parameters. The catalyst was further explored for Knoevenagel condensation reaction, in which it showed its effectiveness and recyclability towards the formation of benzylidenemalononitrile derivatives of aryl aldehydes. Thus, it is a potential 'green catalyst' derived from waste biomass without any addition of chemicals that can replace the industrial base catalysts used for biodiesel production and Knoevenagel reaction and makes the protocol environmentally benign.


Ultrasensitive bacterial sensing using a disposable all-in-one amperometric platform with self-noise cancellation.
The early detection of very low bacterial concentrations is key to minimize the healthcare and safety issues associated with microbial infections, food poisoning or water pollution. In amperometric integrated circuits for electrochemical sensors, flicker noise is still the main bottleneck to achieve ultrasensitive detection with small footprint, cost-effective and ultra-low power instrumentation. Current strategies rely on autozeroing or chopper stabilization causing negative impacts on chip size and power consumption. This work presents a 27-μW potentiostatic-amperometric Delta-Sigma modulator able to cancel its own flicker noise and provide a 4-fold improvement in the limit of detection. The 2.3-mm2 all-in-one CMOS integrated circuit is glued to an inkjet-printed electrochemical sensor. Measurements show that the limit of detection is 15 pArms, the extended dynamic range reaches 110 dB and linearity is R2 = 0.998. The disposable device is able to detect, in less than 1h, live bacterial concentrations as low as 102 CFU/mL from a 50-μL droplet sample, which is equivalent to 5 microorganisms.


Enhanced clearance of vancomycin by hemodialysis in a child.
Clearance of vancomycin by hemodialysis has previously been reported to be negligible. We describe a child undergoing chronic hemodialysis using a hollow fiber dialyzer with cellulose acetate membrane. When the patient was treated with intravenous vancomycin for staphylococcal bacteremia, the serum vancomycin half-life was found to decrease by more than 90% during each course of hemodialysis. We conclude that, contrary to prevailing opinion, vancomycin can be rapidly cleared by hemodialysis in the small pediatric patient, and that the dosage should be adjusted accordingly.


A parathyroid-related peptide induces transcaltachia (the rapid, hormonal stimulation of intestinal Ca2+ transport).
PTH-related peptide (PTHrP) has been shown to be responsible for the hormonal hypercalcemia of malignancy. Previously, we demonstrated that both 1,25(OH)2-vitamin D3 and the N-terminal fragment of PTH (1-34) stimulates the rapid transport of Ca2+ (transcaltachia) in the perfused chick intestine. Since there is a sequence homology between these two hormones in the n-terminal fragment, in this study we examined the effect of PTHrP(1-40) on stimulation of transcaltachia in the perfused chick duodenum. The results indicate that the maximal stimulation of transcaltachia occurs at 50 pM PTHrP(1-40), and that the dose-response curve is biphasic in nature. Perfusion with 25 pM, 50 pM, 100 pM or 200 pM PTHrP(1-40) for 40 min increases the transport of Ca2+ in perfused intestine 1.8-, 3.0-, 2.4- and 1.6-fold, respectively. The response is rapid, occurring within 10 min of introduction of the PTHrP. The Ca2+ channel inhibitor nifedipine, which is known to abolish the transcaltachic effect elicited by 1,25(OH)2 vitamin D3, also inhibited the rapid transport of Ca2+ stimulated by PTHrP(1-40). The transcaltachic effect of PTHrP(1-40) may be mediated by a signal transduction pathway in which Ca2+ channels are activated.


Bifidobacterium breve CBT BR3 is effective at relieving intestinal inflammation by augmenting goblet cell regeneration.
Bifidobacterium breve was the first bacteria isolated in the feces of healthy infants and is a dominant species in the guts of breast-fed infants. Some strains of B. breve have been shown to be effective at relieving intestinal inflammation, but the modes of action have yet to be elucidated. In this study, we investigated the mechanisms of action of B. breve CBT BR3 isolated from South Korean infant feces in relieving colitis in vitro and in vivo. Colitis was induced in mice with dextran sodium sulfate (DSS) and dinitrobenzene sulfonic acid (DNBS). Quantitative reverse-transcription polymerase chain reaction, in vitro FITC-dextran flux permeability assay, and aryl hydrocarbon receptor (AhR) luciferase assay are performed using Caco-2 cells and HT29-Lucia™ AhR cells. B. breve CBT BR3 was orally administered. B. breve CBT BR3 improved colitis symptoms in both DSS- and DNBS-induced colitis models. B. breve CBT BR3 increased the number of goblet cells per crypt. B. breve increased the mRNA expressions of Notch, Spdef, Muc5, and Il22. The mRNA expressions of Occludin, which encodes a membrane tight-junction protein, and Foxo3, which encodes a protein related to butyrate metabolism, were also increased in the DSS- and DNBS-induced colitis models. B. breve CBT BR3 protected inflammation-induced epithelial cell permeability and improved goblet cell function by inducing aryl hydrocarbon receptor in vitro. These results indicate that B. breve CBT BR3 is effective at relieving intestinal inflammation by augmenting goblet cell regeneration.


Structure determination of molecules in an alignment laser field by femtosecond photoelectron diffraction using an X-ray free-electron laser.
We have successfully determined the internuclear distance of I2 molecules in an alignment laser field by applying our molecular structure determination methodology to an I 2p X-ray photoelectron diffraction profile observed with femtosecond X-ray free electron laser pulses. Using this methodology, we have found that the internuclear distance of the sample I2 molecules in an alignment Nd:YAG laser field of 6 × 1011 W/cm2 is elongated by from 0.18 to 0.30 Å "in average" relatively to the equilibrium internuclear distance of 2.666 Å. Thus, the present experiment constitutes a critical step towards the goal of femtosecond imaging of chemical reactions and opens a new direction for the study of ultrafast chemical reaction in the gas phase.


Development of a Prognostic Nomogram for Predicting the Probability of Nonresponse to Total Knee Arthroplasty 1 Year After Surgery.
Indications for total knee arthroplasty (TKA) currently depend on clinical judgment. Up to one fifth of those who undergo primary TKA do not report a clinically meaningful improvement in pain and function after surgery. Our aim was to develop and internally validate a prognostic tool for predicting the probability of nonresponse to surgery at 12 months. Patients from 1 center who underwent primary TKA (N = 615) between 2012 and 2013. The Western Ontario and McMaster Universities Arthritis Index was collected pre- and 12 months after TKA from which nonresponse to surgery was determined using the Outcome Measures in Rheumatology-Osteoarthritis Research Society International responder criteria. Using independent prognostic correlates of postoperative nonresponse observed in adjusted modeling, we derived a prognostic nomogram to estimate the probability of nonresponse to TKA based on this suite of explanatory variables. A total of 90/615 (15%) cases were nonresponders to TKA. The degree of contribution (odds ratio, 95% confidence interval) of each explanatory factor to nonresponse nomogram points was body mass index ≥40 kg/m(2) (3.48; 1.97-6.12), Kellgren and Lawrence <4 (2.59; 1.58-4.24), mental disability on Short Form Health Survey (SF-12) mental component score (3.30; 1.44-7.58), and every 10-point increase in preoperative Western Ontario and McMaster Universities Arthritis Index score (0.81; 0.68-0.97). The concordance index for this model was 0.74. We have created a prognostic nomogram that displays the predictive probabilities of nonresponse to TKA as a source of decision support for clinicians and patients, about their likely functional outcome from TKA. Although our own internal validation suggested good nomogram performance, external validation in a comparable surgical population is required to confirm generalizability of the nomogram.


Centrosome de-clustering of cancer cells induces cGAS-STING-mediated innate immunity of tumor-associated tumor cells in response to irradiation.
Although radiotherapy (RT) increases the extra centrosomes of cancer cells compared to normal cells, centrosome clustering of cancer cells with amplified centrosomes ensures bipolar mitosis for cell proliferation in response to RT. Recent evidence suggests that centrosome clustering is a tumor-selective target for improving RT in breast cancer cells. However, whether centrosome de-clustering is involved in the activation of innate immunity in response to RT remains unknown. In this study, we showed that centrosome de-clustering of irradiated cancer cells modulates cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING)-mediated innate immunity in monocytes and macrophages after co-culture. Centrosome de-clustering intensifies mitotic abnormalities and cytosolic dsDNA in breast cancer cells in response to irradiation. Unexpectedly, centrosome de-clustering did not modulate the cGAS-STING signaling pathway in irradiated breast cancer cells. Importantly, centrosome de-clustering activated the cGAS-STING signaling pathway in human monocytes and mouse macrophages after co-culture with irradiated breast cancer cells. Thus, our data provide the first evidence that centrosome de-clustering of irradiated breast cancer cells induces innate immunity in tumor-associated immune cells.


Predominant phagocytic activity of resident microglia over hematogenous macrophages following transient focal cerebral ischemia: an investigation using green fluorescent protein transgenic bone marrow chimeric mice.
Activated microglia and hematogenous macrophages are known to be involved in infarct development after cerebral ischemia. Traditionally, hematogenic macrophages are thought to be the primary cells to remove the ischemic cell debris. However, phagocytosis is a well known property also of activated microglia. Due to a lack of discriminating cellular markers, the cellular origin of phagocytes and the temporal course of phagocytosis by these two cell types are largely unknown. In this study, we used green fluorescent protein (GFP) transgenic bone marrow chimeric mice and semithin serial sections after methyl methacrylate embedding of the brains to dissect in detail the proportion of identified activated resident microglial cells and infiltrating hematogenous macrophages in phagocytosing neuronal cell debris after 30 min of transient focal cerebral ischemia. Already at day one after reperfusion, we found a rapid decrease of neurons in the ischemic tissue reaching minimum numbers at day seven. Resident GFP-negative microglial cells rapidly became activated at day one and started to phagocytose neuronal material. By contrast, hematogenous macrophages incorporating neuronal cell debris were observed in the ischemic area not earlier than on day four. Quantitative analysis showed maximum numbers of phagocytes of local origin within 2 days and of blood-borne macrophages on day four. The majority of phagocytes in the infarct area were derived from local microglia, preceding and predominating over phagocytes of hematogenous origin. This recruitment reveals a remarkable predominance of local defense mechanisms for tissue clearance over immune cells arriving from the blood after ischemic damage.


Ghrelin improves endothelial function and reduces blood pressure in Ang II-induced hypertensive mice: Role of AMPK.
Endothelial dysfunction is a major pathophysiology observed in hypertension. Ghrelin, a key regulator of metabolism, has been shown to play protective roles in cardiovascular system. However, whether it has the effect of improving endothelial function and lowering blood pressure in Ang II-induced hypertensive mice remains unclear. In this study, hypertension was induced by continuous infusion of Ang II with a subcutaneous osmotic pumps and ghrelin (30 μg/kg/day) was intraperitoneal injection for 4 weeks. Acetylcholine-induced endothelium-dependent relaxation in aortae was measured on wire myograph and superoxide production in mouse aortae was assessed by fluorescence imaging. We found that ghrelin had protective effects on Ang II-induced hypertension by inhibiting oxidative stress, increasing NO production, improving endothelial function, and lowering blood pressure. Furthermore, ghrelin activated AMPK signaling in Ang II-induced hypertension, leading to inhibition of oxidative stress. Compound C, a specific inhibitor of AMPK, reversed the protective effects of ghrelin on the reduction of oxidative stress, the improvement of endothelial function and the reduction of blood pressure. our findings indicated that ghrelin protected against Ang II-induced hypertension by improving endothelial function and lowering blood pressure partly through activating AMPK signaling. Thus, ghrelin may be a valuable therapeutic strategy for hypertension.


Assay of different photoproducts after UVA, B and C irradiation of DNA and human skin explants.
A 32P-HPLC method was applied to study the induction of UVB- and UVC-induced DNA lesions (cyclobutane dimers, 6-4 photoproducts and Dewar isomers) in human skin explants. The employed technique was sensitive enough to detect the lesions at a dose of 10 J/m2 UVB. Comparison of photoadduct formation under UVC and UVB indicated the importance of photosensitization pathways in DNA damage. Dewar isomers were detected only at a high dose of UVB. The compounds were identified by their photochemical reactivity and by spiking with prepared standards. Treatment with nuclease P1 was used to identify the 5'-terminal nucleotide. UVA caused no detectable adducts.


Comparative foraging behavior of Neotropical robber flies (Diptera: Asilidae).
Robber fly species within a Panamanian rain forest comprised distinct shade-seeking (SS) and light-seeking (LS) groups. Thoracic temperatures of LS species averaged 9.2°C greater than ambient, whereas those of SS species averaged only 1.3°C above ambient. Among SS species, attack rate decreased with increasing body mass, whereas relocation rate and attack and relocation distances increased with increasing body mass. Attack and relocation distances of LS species were similar to those of SS species of similar size. Large (>100 mg) LS species, however, had much higher attack and relocation rates than large SS species. The potential costs and benefits of basking are briefly discussed.


The duplex character of the corticofugal pathway from the striate cortex to the lateral geniculate complex of the cat.
From studies on signal conduction times in the corticofugal pathway from the striate cortex of the cat it has been proposed that S cells project to the LGN and C cells to the PGN. This concept has not received support from the histological examination of axons projecting to the two nuclei. The physiological experiments pointed to a clear segregation not apparent in the distribution of axonal diameters. A larger sample of signal conduction times, collected in the present study, is more consistent with the findings on the morphology of the pathway.


Rapid and sensitive HPLC assay for simultaneous determination of procaine and para-aminobenzoic acid from human and rat liver tissue extracts.
A sensitive and rapid high-performance liquid chromatography method has been developed for simultaneous determination of procaine and its metabolite p-aminobenzoic acid (PABA) from human and rat liver tissue extracts. The method has been validated according to ICH guidelines in terms of selectivity, linearity, lower limit of detection, lower limit of quantitation, accuracy, precision and recovery from human and rat liver tissue extracts. Chromatography was carried out on a Discovery C(18) column using 10mM ammonium acetate at pH 4.0 and acetonitrile as mobile phase. Retention times for procaine and PABA were 6.6 and 5.3 min, respectively. Linearity for each calibration curve in both tissue extracts was observed across a range from 10 microM to 750 microM for procaine and PABA. The lower limit of detection for both procaine and PABA was 5 microM and the lower limit of quantitation was 10 microM in both tissue extracts. The intra- and inter-day relative standard deviations (R.S.D.) for both procaine and PABA were <6%. Recoveries of procaine and PABA from human and rat liver tissue extracts were determined by two different methods with a single-step protein precipitation technique being employed in both methods. Recoveries for both procaine and PABA were greater than 80% from both human and rat liver tissue extracts.


Interaction of Cholera Toxin B-subunit with Human T-lymphocytes.
In this work, 125I-labeled cholera toxin B-subunit (CT-B) (specific activity 98 Ci/mmol) was prepared, and its high-affinity binding to human blood T-lymphocytes (Kd = 3.3 nM) was determined. The binding of the 125I-labeled CT-B was inhibited by unlabeled interferon-α2 (IFN-α2), thymosin-α1 (TM-α1), and by the synthetic peptide LKEKK, which corresponds to sequences 16-20 of human TM-α1 and 131-135 of IFN-α2 (Ki 0.8, 1.2, and 1.6 nM, respectively), but was not inhibited by the unlabeled synthetic peptide KKEKL with inverted sequence (Ki > 1 µM). In the concentration range of 10-1000 nM, both CT-B and peptide LKEKK dose-dependently increased the activity of soluble guanylate cyclase (sGC) but did not affect the activity of membrane-bound guanylate cyclase. The KKEKL peptide tested in parallel did not affect sGC activity. Thus, the CT-B and peptide LKEKK binding to a common receptor on the surface of T-lymphocytes leads to an increase in sGC activity.


Immunoregulation by lymphokines: immune interferon and lymphotoxin induction of lymphokine activity in human peripheral blood leukocyte cultures.
Supernatants of mitogen-induced human PBL cultures contain several lymphokine activities. Analysis of kinetic studies has suggested a correlation in the appearance in culture supernatants of lymphotoxin (LT) and immune interferon (Im-IF). We investigated this interaction between LT and Im-IF in the serum-free human PBL culture system. Partially purified LT was derived from a normal cell line, RPMI 1788. This LT, which had no contaminating IF activity, induced production of significant levels of IF activity as early as 15 hours after initiation of culture. This induced IF activity was identified as Im-IF by criteria of pH 2, SDS and heat sensitivity, as well as neutralization by AB to Im-IF. LT activity, in excess of the units added to the culture, was also measured in the LT-induced cultures. In another group of experiments, partially purified human Im-IF, produced by expression of a cloned gene in monkey COS-7 cells, was added to human PBL cultures. This Im-IF, at concentrations of 160 and 320 units/ml, caused induction of LT activity. The induced LT activity was neutralized by AB raised to 1788-derived LT. Im-IF treated cultures also contained IF activity in excess of the titers added to the cultures. These results would suggest an interaction between LT and Im-IF at the level of production by lymphoid cells.


Hiring Requirements and Qualifications of School Food Authority Directors Changed in Some Districts After Implementation of US Department of Agriculture Professional Standards.
In 2015, the US Department of Agriculture set minimum education and training requirements (ie, professional standards) to ensure that school nutrition professionals have the knowledge and experience to operate school meal programs. No study to date has examined whether hiring requirements and qualifications of school food authority (SFA) directors have changed since 2015. To assess changes in hiring requirements and qualifications of SFA directors since the US Department of Agriculture professional standards were established, overall and by district size. Cross-sectional analysis of nationally representative district-level data from the 2012 and 2016 cycles of the School Health Policies and Practices Study. In 2012, 660 sampled districts completed the School Health Policies and Practices Study Nutrition Services questionnaire. In 2016, 599 sampled districts completed the questionnaire. Hiring requirements for newly hired SFA directors and reported qualifications of SFA directors. Differences in prevalence estimates from 2012 to 2016 for all districts and by district size were assessed with χ2 tests. Significant increases were found for 3 hiring requirements: degree in nutrition or related field, registered dietitian credential, and food safety certification. Significant changes in 4 of the 5 reported qualifications were found including an increase in the percentage of district directors with a degree in nutrition or a related field and decreases in the percentage of directors with a School Nutrition Specialist credential from the School Nutrition Association, School Nutrition Association certifications, and certified dietary managers. Changes were found in small and medium districts, but not large districts. District hiring requirements and SFA director qualifications have changed since the implementation of the US Department of Agriculture professional standards, including some differences by district size. Future research could identify challenges facing districts in hiring directors who have a degree in nutrition or related fields or who have specialized nutrition credentials or certificates (eg, registered dietitians).


Responses to drugs of abuse and non-drug rewards in leptin deficient ob/ob mice.
Although leptin receptors are found in hypothalamic nuclei classically associated with homeostatic feeding mechanisms, they are also present in brain regions known to regulate hedonic-based feeding, natural reward processing, and responses to drugs of abuse. The ob/ob mouse is deficient in leptin signaling, and previous work has found altered mesolimbic dopamine signaling and sensitivity to the locomotor activating effects of amphetamine in these mice. We directly assessed responses to three drugs of abuse and non-drug rewards in the leptin-deficient ob/ob mouse. Ob/ob mice were tested in assays of sweet preference, novelty seeking, and drug reward/reinforcement. In assays of novelty seeking, novel open field activity and operant sensation seeking were reduced in ob/ob mice, although novel object interaction and novel environment preference were comparable to wild types. We also found that ob/ob mice had specific phenotypes in regard to cocaine: conditioned place preference for 2.5 mg/kg was increased, while the locomotor response to 10 mg/kg was reduced, and cocaine self-administration was the same as wild types. Ob/ob mice also acquired self-administration of the potent opioid remifentanil, but breakpoints for the drug were significantly reduced. Finally, we found significant differences in ethanol drinking in ob/ob mice that correlated negatively with body weight and positively with operant sensation seeking. In conclusion, ob/ob mice displayed task-specific deficits in novelty seeking and dissociable differences in reward/reinforcement associated with cocaine, remifentanil, and ethanol.