Metabolic adaptation to vitamin auxotrophy by leaf-associated bacteria.
Auxotrophs are unable to synthesize all the metabolites essential for their metabolism and rely on others to provide them. They have been intensively studied in laboratory-generated and -evolved mutants, but emergent adaptation mechanisms to auxotrophy have not been systematically addressed. Here, we investigated auxotrophies in bacteria isolated from Arabidopsis thaliana leaves and found that up to half of the strains have auxotrophic requirements for biotin, niacin, pantothenate and/or thiamine. We then explored the genetic basis of auxotrophy as well as traits that co-occurred with vitamin auxotrophy. We found that auxotrophic strains generally stored coenzymes with the capacity to grow exponentially for 1-3 doublings without vitamin supplementation; however, the highest observed storage was for biotin, which allowed for 9 doublings in one strain. In co-culture experiments, we demonstrated vitamin supply to auxotrophs, and found that auxotrophic strains maintained higher species richness than prototrophs upon external supplementation with vitamins. Extension of a consumer-resource model predicted that auxotrophs can utilize carbon compounds provided by other organisms, suggesting that auxotrophic strains benefit from metabolic by-products beyond vitamins.


Vitamin E, nuclear receptors and xenobiotic metabolism.
Supplemental vitamin E (alpha-tocopherol) is taken daily by more than 35 million people in the US. Following absorption and liver uptake, the fate of vitamin E is largely unknown. Of potential importance are recent clinical studies that have reported adverse effects of vitamin E that may be directly related to its hepatic metabolism. In an in vitro system, both vitamin E and rifampicin, a known stimulator of xenobiotic metabolism, activated the pregnane X receptor (PXR), an orphan nuclear receptor. PXR as a heterodimer with the retinoid X receptor (RXR), binds to specific cis-elements in the promoter regions of genes. PXR/RXR regulates a constellation of genes involved in xenobiotic detoxification, including oxidation, conjugation, and transporters. Importantly, PXR/RXR regulates the cytochrome P450 (CYP), CYP3A, involved in the hepatic detoxification of more than 50% of prescription drugs. Vitamin E acting as a PXR ligand could alter these PXR-mediated reactions. Unfortunately, the extent to which pharmacologic doses of vitamin E modulate these pathways in vivo has not been determined.


Ultrastructure of the osphradium of Aplysia californica.
The osphradium of Aplysia californica, a sensory organ, is a small yellow-brown epithelial patch located in the mantle cavity immediately anterior to the rostral attachment of the gill. Scanning electron microscopy reveals a round ellipsoid structure of 0.6-1 mm in diameter with a central, occasionally folded, sensory epithelium. The central area is covered with microvilli and surrounded by a densely ciliated epithelium. Transmission electron micrographs show that the columnar supporting cells in the sensory epithelium contain an abundance of apical pigment granules and microvilli. Between the epithelial-supporting cells, the putative sensory elements consist of thin neurites (0.4-1.5 micron in diameter) that reach the sea-water side of the osphradium. The neurites contain many neurotubules, mitochondria, vesicles and cilia in their apices. The nerve endings originate from cell bodies up to 40 microns below the epithelium or in the osphradial ganglion itself, as revealed by electron microscopy and retrograde labeling with Lucifer yellow. There appear to be two populations of putative sensory cells, a large population of heavily stained cell bodies 4-10 microns in diameter and a few scattered cells of large diameter (25-60 microns). Following lanthanum impregnation, septate junctions can be seen between all types of cells in the epithelium, 3-5 microns below the sea-water surface. This study provides new information for further investigation of osmo- and mechanosensation in Aplysia californica.


Primary care nursing competencies in Canadian undergraduate nursing programs: A national cross-sectional survey.
To assess the extent to which Canadian undergraduate baccalaureate nursing programs have incorporated Canadian competencies for Registered Nurses in primary care into their curricula. Canadian competencies for Registered Nurses in primary care have several benefits, including their ability to inform primary care education in undergraduate nursing programs and to assist in building a robust primary care nursing workforce. We conducted a national cross-sectional survey of undergraduate baccalaureate nursing programs (n = 74). The survey was conducted between April-May 2022. We used a modified version of the "Community Health Nurses' Continuing Education Needs Questionnaire". Respondents indicated their level of agreement on a 6-point Likert scale with 47 statements about the integration of the competencies in their program (1 = strongly disagree; 6 = strongly agree). The response rate was 51.4%. The overall mean across the six competency domains was 4.73 (SD 0.30). The mean scores of each domain ranged from 4.23 (SD 1.27) for Quality Assurance, Evaluation and Research to 5.17 (SD 0.95) for Communication. There are gaps in how these competencies are included in undergraduate education programs and opportunities to strengthen education for this growing workforce in Canada.


Influence of Risk Factors on the Well-Being of Elderly Women with Knee Osteoarthritis.
Background and Objectives: Knee osteoarthritis (KOA) is a widespread chronic joint disease characterized by functional limitations and pain. Functioning restrictions exert a detrimental impact on societal integration, relationships, and psychological well-being, resulting in significant emotional distress in KOA patients. The objective of this study is to examine how various risk factors impact the emotional well-being of individuals with KOA. Materials and Methods: This prospective cross-sectional study involved 154 postmenopausal women treated at the Special Hospital for Rheumatic Diseases in Novi Sad, Serbia. The experimental group comprised 97 individuals with chronic knee pain and structural knee damage (Kellgren-Lawrence (KL) scale II-IV), while the control group had 53 individuals with chronic knee pain but no structural knee damage (KL scale 0-I). The collected data consisted of sociodemographic factors, general characteristics, associated diseases, and laboratory results. Adequate anthropometric measurements were conducted, and all subjects were required to complete the SF-36 RAND questionnaire. Results: The analysis identified several variables that independently influenced emotional well-being. These included pain intensity (beta (β) 0.21; 95% CI: 0.03-0.20; p < 0.01), social functioning (beta (β) 0.47; 95% CI: 0.23-0.43; p < 0.001), physical functioning (beta (β) 0.23; 95% CI: 0.04-0.21; p < 0.01), and education level (8-12 years: beta (β) 0.25; 95% CI: 1.47-9.41; p < 0.01; >12 years: beta (β) 0.27; 95% CI: 2.51-12.67; p < 0.01). However, the multivariate model revealed that only social functioning (beta (β) 0.57; 95% CI: 0.27-0.53; p < 0.001) and education level (8-12 years: beta (β) 0.21; 95% CI: 1.10-8.260; p < 0.05; >12 years: beta (β) 0.21; 95% CI: 1.18-10.30; p < 0.05) were significantly associated with emotional well-being in KOA patients. Conclusions: The findings of this study indicate that a reduced social functioning and a lower educational attainment are linked to a poorer emotional well-being among patients with KOA.


Impact of neck percutaneous interferential current sensory stimulation on swallowing function in patients with Parkinson's disease: A single-arm, open-label study protocol.
Parkinson's disease (PD) can lead to swallowing dysfunction, resulting in aspiration pneumonia. Among the types of swallowing disorders, a characteristic and serious problem associated with PD is silent aspiration due to pharyngeal and laryngeal hypoesthesia. This single-arm, open-label study aims to evaluate the effectiveness of percutaneous neck interferential current sensory stimulation in enhancing swallowing function in patients with PD. The efficacy and safety of percutaneous neck interferential current sensory stimulation will be investigated for patients diagnosed with PD, based on the Movement Disorder Society criteria, of Hoehn-Yahr stages 2-4. The patients will receive neck percutaneous interferential current sensory stimulation for 20 min twice a week for 8 weeks using a Gentle Stim® (FoodCare Co., Ltd., Kanagawa, Japan) device. Once the intervention is initiated, evaluations will be performed every 4 weeks for a 16-week period. The primary endpoint to be assessed is the proportion of patients with normal cough with 1% citric acid at the end of the intervention (8 weeks after intervention initiation) compared with that at the beginning. This clinical trial will examine the usefulness of percutaneous neck interferential current sensory stimulation in patients with PD. In addition, this study will use novel instruments, such as multichannel surface electromyography and electronic stethoscope, to evaluate swallowing function. This novel evaluation can provide insights into dysphagia in patients with PD and the usefulness of percutaneous neck interferential current stimulation. This exploratory study is limited by its single-arm, open-label design and small size. jRCTs062220013; pre-results.


A stress-blinded Atf1 can fully assemble heterochromatin in a RNAi-independent minimal mat locus but impairs directionality of mat2/3 switching.
The MAP kinase Sty1 phosphorylates and activates the transcription factor Atf1 in response to several stress conditions, which then shifts from a transcriptional repressor to an activator. Atf1 also participates in heterochromatin assembly at the mat locus, in combination with the RNA interference (RNAi) machinery. Here, we study the role of signal-dependent phosphorylation of Atf1 in heterochromatin establishment at mat, using different Atf1 phospho mutants. Although a hypo-phosphorylation Atf1 mutant, Atf1.10M, mediates heterochromatin assembly, the phosphomimic Atf1.10D is unable to maintain silencing. In a minimal mat locus, lacking the RNAi-recruiting cis elements and displaying intermediate silencing, Atf1.10M restores full heterochromatin and silencing. However, evolution experiments with this stress-blinded Atf1.10M show that it is unable to facilitate switching between the donor site mat3 and mat1. We propose that the unphosphorylated, inactive Atf1 contributes to proper heterochromatin assembly by recruiting repressive complexes, but its stress-dependent phosphorylation is required for recombination/switching to occur.


Intervention with hand orthosis: experience from boys with Duchenne muscular dystrophy and their parents.
To identify and describe possible ways of experiencing an intervention with hand orthoses in a study group including boys with Duchenne muscular dystrophy (DMD) and their parents. To capture the experiences of the persons directly involved as well as their family members. Eight boys with DMD (aged 8-21; median age 17.7) and five of their parents were interviewed immediately post-intervention. Additionally, follow-up interviews were later performed with five boys and three parents. Ten categories of intervention experiences emerged and were allocated to three aspects: "Prerequisites in the treatment situation", "The intervention makes a difference" and "Instilling hope for the future". The requirements for intervention success include co-operation with parents and hand orthoses with a good fit. Maintained or increased joint mobility, reduced pain and improved occupational performance were experienced. The boys and parents perceived that the intervention with hand orthoses could counteract the deterioration of the boys' hands. This instilled hope of preserving occupational performance throughout life. They also considered that a good hand-orthosis fit, appropriate adjustments to daily routines and good co-operation with the people around them were important for the intervention to be and remain successful.


The glucocorticoid RU24858 does not distinguish between transrepression and transactivation in primary human eosinophils.
Glucocorticoids are used to treat chronic inflammatory diseases such as asthma. Induction of eosinophil apoptosis is considered to be one of the main mechanisms behind the anti-asthmatic effect of glucocorticoids. Glucocorticoid binding to its receptor (GR) can have a dual effect on gene transcription. Activated GR can activate transcription (transactivation), or by interacting with other transcription factors such as NF-kappaB suppress transcription (transrepression). RU24858 has been reported to transrepress but to have little or no transactivation capability in other cell types. The dissociated properties of RU24858 have not been previously studied in non-malignant human cells. As the eosinophils have a very short lifetime and many of the modern molecular biological methods cannot be used, a "dissociated steroid" would be a valuable tool to evaluate the mechanism of action of glucocorticoids in human eosinophils. The aim of this study was to elucidate the ability of RU24858 to activate and repress gene expression in human eosinophils in order to see whether it is a dissociated steroid in human eosinophils. Human peripheral blood eosinophils were isolated under sterile conditions and cultured in the presence and/or absence RU24858. For comparison, dexamethasone and mometasone were used. We measured chemokine receptor-4 (CXCR4) and Annexin 1 expression by flow cytometry and cytokine production by ELISA. Apoptosis was measured by DNA fragmentation and confirmed by morphological analysis. RU24858 (1 microM) increased CXCR4 and Annexin 1 expression on eosinophils to a similar extent as mometasone (1 microM) and dexamethasone (1 microM). Like dexamethasone and mometasone, RU24858 did suppress IL-8 and MCP-1 production in eosinophils. RU24858 also increased spontaneous eosinophil apoptosis to a similar degree as dexamethasone and mometasone, but unlike dexamethasone and mometasone it did not reverse IL-5- or GM-CSF-induced eosinophil survival. Our results suggest that in human eosinophils RU24858 acts as transactivator and transrepressor like classical glucocorticoids. Thus, RU24858 seems not to be a "dissociated steroid" in primary human eosinophils in contrast to that reported in animal cells. In addition, functionally RU24858 seems to be a less potent glucocorticoid as it did not reverse IL-5- and GM-CSF-afforded eosinophil survival similarly to dexamethasone and mometasone.


Microemulsions as transdermal drug delivery vehicles.
Microemulsions are clear, stable, isotropic mixtures of oil, water, and surfactant, frequently in combination with a cosurfactant. Microemulsions have been intensively studied during the last decades by many scientists and technologists because of their great potential in many food and pharmaceutical applications. The use of microemulsions is advantageous not only due to the facile and low cost preparation, but also because of the improved bioavailability. The increased absorption of drugs in topical applications is attributed to enhancement of penetration through the skin by the carrier. Saturated and unsaturated fatty acids serving as an oil phase are frequently used as penetration enhancers. The most popular enhancer is oleic acid. Other permeation enhancers commonly used in transdermal formulations are isopropyl myristate, isopropyl palmitate, triacetin, isostearylic isostearate, R(+)-limonene and medium chain triglycerides. The most popular among the enhancing permeability surfactants are phospholipids that have been shown to enhance drug permeation in a different mode. l-alpha-phosphatidylcholine from egg yolk, l-alpha-phosphatidylcholine 60%, from soybean and dioleylphosphatidyl ethanolamine which are in a fluid state may diffuse into the stratum corneum and enhance dermal and transdermal drug penetration, while distearoylphosphatidyl choline which is in a gel-state has no such capability. Other very commonly used surfactants are Tween 20, Tween 80, Span 20, Azone, Plurol Isostearique and Plurol Oleique. As cosurfactants commonly serve short-chain alkanols such as ethanol and propylene glycol. Long-chain alcohols, especially 1-butanol, are known for their enhancing activity as well. Decanol was found to be an optimum enhancer among other saturated fatty alcohols that were examined (from octanol to myristyl alcohol). Many enhancers are concentration-dependent; therefore, optimal concentration for effective promotion should be determined. The delivery rate is dependent on the type of the drug, the structure and ingredients of the carrier, and on the character of the membrane in use. Each formulation should be examined very carefully, because every membrane alters the mechanism of penetration and can turn an enhancer to a retarder. Various potential mechanisms to enhance drug penetration through the skin include directly affecting the skin and modifying the formulation so the partition, diffusion, or solubility is altered. The combination of several enhancement techniques such as the use of iontophoresis with fatty acids leads to synergetic drug penetration and to decrease in skin toxicity. Selected studies of various microemulsions containing certain drugs including retinoic acid, 5-fluorouracil, triptolide, ascorbic acid, diclofenac, lidocaine, and prilocaine hydrochloride in transdermal formulations are presented in this review. In conclusion, microemulsions were found as an effective vehicle of the solubilization of certain drugs and as protecting medium for the entrapped of drugs from degradation, hydrolysis, and oxidation. It can also provide prolonged release of the drug and prevent irritation despite the toxicity of the drug. Yet, in spite of all the advantages the present formulations lack several key important characteristics such as cosmetic-permitted surfactants, free dilution in water capabilities, stability in the digestive tracts and sufficient solubilization capacity.


Beyond oxidative stress: an immunologist's guide to reactive oxygen species.
Reactive oxygen species (ROS) react preferentially with certain atoms to modulate functions ranging from cell homeostasis to cell death. Molecular actions include both inhibition and activation of proteins, mutagenesis of DNA and activation of gene transcription. Cellular actions include promotion or suppression of inflammation, immunity and carcinogenesis. ROS help the host to compete against microorganisms and are also involved in intermicrobial competition. ROS chemistry and their pleiotropy make them difficult to localize, to quantify and to manipulate - challenges we must overcome to translate ROS biology into medical advances.


A monoclonal antibody-based VZV glycoprotein E quantitative assay and its application on antigen quantitation in VZV vaccine.
Varicella-zoster virus (VZV) is a highly infectious agent that causes varicella and herpes zoster (HZ), which may be associated with severe neuralgia. Vaccination is the most effective way to reduce the burden of the diseases. VZV glycoprotein E (gE) is the major and most immunogenic membrane protein that plays important roles in vaccine efficacy. A quantitative assay for gE content is desirable for the VZV vaccine process monitoring and product analysis. In this study, 70 monoclonal antibodies (mAbs) were obtained after immunizing mice with purified recombinant gE (rgE). The collection of mAbs was well-characterized, and a pair of high-affinity neutralization antibodies (capture mAb 4A2 and detection mAb 4H10) was selected to establish a specific and sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to quantify the native and recombinant gE. The detection limit of this assay was found to be 1.95 ng/mL. Furthermore, a reasonably good correlation between the gE content (as measured by the mAb-based quantitative ELISA) and the virus titer (as measured by the "gold standard" plaque assay) was observed when both assays were performed for tracking the kinetics of virus growth during cell culture. A total of 16 batches of lyophilized VZV vaccine were tested using the newly developed quantitative ELISA and classical plaque assay, demonstrating reasonably good correlation between gE content and virus titer. Therefore, this mAb-based gE quantitative assay serves as a rapid, stable, and sensitive method for monitoring viral antigen content, one additional quantitative method for VZV vaccine process and product characterization. This quantitative ELISA may also serve as a complementary method for virus titering.


Hyperinsulinemia in myotonic dystrophy: identity of the maternal factor causing the neonatal myotonic dystrophy syndrome.
An environmental factor acting on the fetus is thought to cause a neonatal syndrome characterized by marked muscular hypotonia, lack of respiratory drive and feeding difficulties, in some infants born to mothers with myotonic dystrophy. Mortality is high, especially amongst those babies born prematurely, but muscle strength and tone improve rapidly in survivors. Nevertheless, most survivors have physical deformities and mental retardation and are thought to develop myotonic dystrophy later. We propose that alterations in maternal insulin secretion (usual in myotonic dystrophy subjects) alter fetal blood glucose and amino acid levels and retard growth and maturation of fetal skeletal muscle. This leads to severe muscular hypotonia in affected infants. Also, we suggest that infants who die during the perinatal period may not have inherited the defective autosomal dominant gene that causes myotonic dystrophy.


The role of iron chelates on the selectivity of Fenton reagent in hydroxylation, N-demethylation, and sulfoxidation of cimetidine: a novel biomimetic model for the regioselectivity of cytochrome P-450.
The effect of iron chelates on the reaction of cimetidine with Fenton reagent [Fe(II)/H2O2] has been investigated. Iron chelates with high affinity to ferrous ions inhibited this reaction. However, iron chelates with high affinity to ferric ions selectively promote either hydroxylation, N-demethylation, or sulfoxidation of cimetidine. These results indicate that the oxidation of cimetidine with hydrogen peroxide activated by various chelated ferrous ions serves as a biomimetic model for the regioselectivity of multiple forms of cytochrome P-450 in the metabolism of cimetidine.


[When all hope has gone…well-placed affection remains].
Cédric de Linage experienced coma through his wife, Amélie, after she suffered cardiopulmonary arrest. Here he describes the crucial role, during this period of uncertainty and fear, played by the nursing team. Through their care and day-to-day compassion, the team was able to treat the patient as a fragile but living patient.


The optimal frequency of endoscopic skill labs for training and skill retention on suturing: a randomized controlled trial.
To determine, given a fixed amount of training, the optimal distribution of sessions needed to acquire intracorporeal endoscopic suturing skills and to retain said skills for 1 to 6 months. A randomized controlled trial consisted of 6 study groups who received identical laparoscopic suturing training but at differing frequencies of sessions. The faculty of medicine of the KULEUVEN is the largest medical faculty in Belgium. Medical students without experience in laparoscopy (n = 145). After baseline assessments, the students were randomized into 6 groups to receive 6 training sessions of 1.5 hours each. Training Groups were as follows: 3 sessions daily (TD), bidaily sessions, 1 session daily (OD), 1 session on alternative days, 1 session weekly, and 1 session weekly with an optional "deliberate practice" in between sessions (WD). All exercises and feedback given were identical. One and 6 months after the final session, an evaluation was performed where a 5-cm chicken-skin incision had to be closed with 3 laparoscopic knots. The cumulative time to approximate the skin edges adequately was used for qualitative and quantitative analysis. There were no significant differences amongst the groups at baseline concerning ambidexterity, motivation, or spatial abilities. The group OD outperformed the massed groups (TD and bidaily sessions) and the weekly groups (1 session weekly and WD) significantly (p = 0.003). After 1 month there was still a significant advantage for regular training groups (OD, 1 session on alternative days, and WD) over massed training groups (TD) (p = 0.004). After 6 months only a statistical difference (p = 0.04) between group OD and group TD was observed. Group WD's score remained stable after 6 months. Short-term, once daily 1.5-hours session seems most beneficial for learning intracorporeal endoscopic suturing. After 6 months, distributed shorter training still remains better than massed practice but weekly training and daily training are comparable. "Optional Deliberate Practice" between the official training sessions reduces skill decay.


Possible implications of the comparative dissociative behavior of testosterone- and 5 alpha-dihydrotestosterone-receptor complexes formed by human skin fibroblasts.
We have studied the dissociative behavior of 5 alpha-dihydrotestosterone (DHT)- or testosterone (T)-receptor complexes extracted from normal and 5 alpha-reductase-deficient human genital skin fibroblasts, respectively. The rate at which each hormone dissociates from the androgen receptor is temperature-dependent (29-40 degrees C), but DHT does so in a first-order (monophasic) manner, while T yields complex (biphasic) dissociation kinetics. Prior exposure to 0.1 M sodium thiocyanate induces DHT-receptor complexes to dissociate with complex kinetics. The temperature dependence of these complex dissociation profiles can be simulated identically by a kinetic model involving three conformational states of the androgen-receptor complex. Neither the rate constants of the rapidly- or slowly-dissociating components of T-receptor dissociation, nor the magnitude of the latter are affected by sodium molybdate (5 mM). Similarly, neither sodium molybdate nor potassium chloride (0.1-0.3 M) alters the complex dissociative behavior of DHT-receptor complexes that is induced by 0.1 M sodium thiocyanate.


Serum quantitative proteomic analysis of patients with keshan disease based on iTRAQ labeling technique: A first term study.
Keshan disease (KD), an endemic myocardiopathy, with unknown etiology, is still threatening human health. Proteomics studies of Keshan disease is rarely known. In the current study, isobaric tags for relative and absolute quantitation (iTRAQ) labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) was firstly used to screen for the differentially expressed proteins (DEPs) in serum between Keshan disease and healthy residents in endemic area. 27 differentially expressed proteins were quantified, 9 of which were significantly altered (fold change > 1.2 or < 0.8) between the two groups. Bioinformatic analysis revealed that a wide variety of biology process played roles in KD pathophysiology. These differentially expressed proteins were mainly involved in complement coagulation pathways. A significant high serum level of LGALS3BP were observed. Our first term study in KD proteomics provided new view into the molecular mechanisms of KD disease.


Immunogold labeling-induced synergy effect for amplified photoelectrochemical immunoassay of prostate-specific antigen.
A new photoelectrochemical immunoassay for prostate-specific antigen (PSA) was successfully developed with high sensitivity via immunogold labeling.


[Regional phlebography in patients with recurrences of varicocele].
Diagnostic potentialities of phlebography were studied in 50 patients with varicocele recurrences. Two methods of phlebography: transfemoral renospermaticography and transscrotal orthograde testiculophlebography--were employed. The main cause of recurrence development was shown to be the abnormality of venous outflow from the testicle in the region of the left renal vein, testicular vein and in the pelvic veins. The effectiveness of both methods was 71.9 and 94.4%, respectively, their combined use provided additional information.


[Cerebral malaria and maternal deaths--a study of 13,141 deliveries at the Maputo Central Hospital, People's Republic of Mozambique].
In the Central Hospital of Maputo we registered 1986 6 maternal deaths during delivery caused by cerebral malaria. Clinical and histopathological findings have been discussed. In endemic districts of malaria convulsions during delivery are hints to cerebral malaria. Following postpartal infections cerebral malaria together with eclampsia has the second position in the reasons of maternal death.


Middle East respiratory syndrome coronavirus: a case-control study of hospitalized patients.
There is a paucity of data regarding the differentiating characteristics of patients with laboratory-confirmed and those negative for Middle East respiratory syndrome coronavirus (MERS-CoV). This is a hospital-based case-control study comparing MERS-CoV-positive patients (cases) with MERS-CoV-negative controls. A total of 17 case patients and 82 controls with a mean age of 60.7 years and 57 years, respectively (P = .553), were included. No statistical differences were observed in relation to sex, the presence of a fever or cough, and the presence of a single or multilobar infiltrate on chest radiography. The case patients were more likely to be overweight than the control group (mean body mass index, 32 vs 27.8; P = .035), to have diabetes mellitus (87% vs 47%; odds ratio [OR], 7.24; P = .015), and to have end-stage renal disease (33% vs 7%; OR, 7; P = .012). At the time of admission, tachypnea (27% vs 60%; OR, 0.24; P = .031) and respiratory distress (15% vs 51%; OR, 0.15; P = .012) were less frequent among case patients. MERS-CoV patients were more likely to have a normal white blood cell count than the control group (82% vs 52%; OR, 4.33; P = .029). Admission chest radiography with interstitial infiltrates was more frequent in case patients than in controls (67% vs 20%; OR, 8.13; P = .001). Case patients were more likely to be admitted to the intensive care unit (53% vs 20%; OR, 4.65; P = .025) and to have a high mortality rate (76% vs 15%; OR, 18.96; P < .001). Few clinical predictors could enhance the ability to predict which patients with pneumonia would have MERS-CoV. However, further prospective analysis and matched case-control studies may shed light on other predictors of infection.


Isoniazid therapy for Mycobacterium tuberculosis infection in HIV clinics, Los Angeles, California.
Publicly funded human immunodeficiency virus (HIV) clinics in Los Angeles County, California, USA. HIV-infected persons are a high priority group for targeted testing and treatment for Mycobacterium tuberculosis infection in the United States. To describe rates of isoniazid (INH) initiation and completion among HIV-1 and M. tuberculosis co-infected persons in Los Angeles County. We conducted a cross-sectional study using routinely collected surveillance data from publicly funded HIV clinics. We examined differences in INH treatment initiation and completion between four clinic categories: the three largest clinics (Clinics A, B, and C) and 'Other' clinics (pooled data for the remaining 10 clinics). During 2010-2013, 802 (5.3%) of 15 029 HIV-1-infected persons tested positive for M. tuberculosis infection. INH was initiated in 581 (72.4%) persons, of whom 457 (78.7%) completed treatment. We found significant differences between clinics in terms of treatment initiation (range 59.1-93.4%) and completion (range 58.8-82.3%). Overall, 57% (457/802) of HIV and M. tuberculosis co-infected persons completed the recommended treatment (range across clinics 34.8-76.3%). We identified significant gaps in the treatment for M. tuberculosis infection among HIV-infected persons in Los Angeles County. Interventions are needed to improve initiation and completion of treatment for M. tuberculosis infection in this population.


Sexual health of Indian immigrant men in Australia: an exploratory research on help-seeking attitudes.
The help-seeking attitudes for sexual health of Indian men living in Australia was explored. Of all survey respondents (n=225), many preferred to seek help from medical doctors. Young (18-25 years) Indian men were three times more likely to prefer a specialist medical doctor than older men. Ethnicity and gender of the medical doctor was 'not important' for the majority of men. Most men preferred to seek help from their regular general practitioner.


Effect of continuous combined estrogen and desogestrel hormone replacement therapy on serum lipids and lipoproteins.
To determine the effects of continuous combined hormone replacement therapy with desogestrel and 17 beta-estradiol (E2) on serum lipids and lipoproteins. Fifty-seven healthy postmenopausal women of less than 60 years of age were studied prospectively and treated with oral desogestrel 0.15 mg/day and micronized 17 beta-E2 1 mg/day, both taken continuously. Fasting venous blood samples for serum lipids and lipoproteins were taken before and after 6 and 12 months of treatment. Thirty-two women completed the study. Levels of all serum lipids and lipoproteins fell significantly by 6 months and remained low at 12 months. The mean percentage reduction after 12 months of treatment was 12.8% for high-density lipoprotein (HDL) cholesterol, which largely resulted from a reduction in the HDL2 subfraction, which fell by 25.7%. The mean percentage reduction for both low-density lipoprotein (LDL) cholesterol and triglycerides was 7.7%. The median percentage reduction for lipoprotein (a) was 17.6%. This combination of hormone replacement therapy had profound effects on serum lipids and lipoproteins. According to current concepts, reductions in total and LDL cholesterol, triglycerides, and lipoprotein (a) may reduce cardiovascular disease risk. The reduction in HDL was unexpected, given the rise in HDL that has been demonstrated when desogestrel is combined with ethinyl estradiol in the contraceptive pill. The lowering of HDL observed in this study is undesirable and may be potentially harmful. Our results indicate that when desogestrel 0.15 mg/day is combined with micronized 17 beta-E2 1 mg/day in a continuous manner, the effects of the progestogen on HDL predominate and cause a reduction in HDL and the HDL2 subfraction.


Characterization of two adenosine 3':5'-phosphate-dependent protein kinase species from Chinese hamster ovary cells.
Chinese hamster ovary cells exhibit several characteristic morphological and physiological responses upon treatment with agents which increase the intracellular level of adenosine 3':5'-phosphate (cyclic AMP). To better understand the mechanism of these cyclic AMP-mediated responses, we separated two cyclic AMP-dependent protein kinases (ATP:protein phosphotransferase, EC 18.104.22.168) (protein kinase I and protein kinase II) from the cytosol of Chinese hamster ovary cells by DEAE-cellulose chromatography and studied their properties. Protein kinase I is eluted at a lower salt concentration than protein kinase II and is stimulable to 10 times its basal catalytic activity, while protein kinase II is stimulable only 2-fold. Both kinases are completely dissociated by cyclic AMP and inhibited by specific cyclic AMP-dependent protein kinase inhibitor. They have similar Km values for magnesium (approximately 1 mM), cyclic AMP (approximately 60 nM), and ATP (approximately 0.1 mM), and the dissociation constant (Kdis) for cyclic AMP (approximately 13 nM) is the same for both enzymes. However, they appear to have different substrate preferences and cyclic AMP-binding properties in that cyclic AMP bound to protein kinase II exchanges readily with free cyclic AMP, while that bound to protein kinase I is not exchangeable. The native enzymes have different sedimentation coefficients (6.4 S for protein kinase I and 4.8 S for protein kinase II), whereas those of the activated enzymes are the same (2.9--3.0 S). It appears that the two cyclic AMP-dependent protein kinases which differ from each other in their regulatory subunits may play different roles in the mediation of cyclic AMP action in Chinese hamster ovary cells.


In silico optimization of merocyanine-spiropyran compounds as second-order nonlinear optical molecular switches.
Time-dependent Hartree-Fock and Møller-Plesset second-order calculations have been used to unravel the relationships between structure and first hyperpolarizability in spiropyran/merocyanine couples and therefore to design efficient second-order nonlinear optical switching compounds. Large first hyperpolarizabilities for the merocyanine form as well as large contrasts of first hyperpolarizability have been obtained when, on the same species, (i) substituents at R(1) and R(2) positions on the phenolate ring of the merocyanine form are strong acceptor and donor substituents, respectively, (ii) the ethylenic bridge is substituted by donor groups, (iii) the other aromatic part of the system is benzimidazolo rather than indolino or benzothiazolo, and (iv) strong donor substituents are placed on the benzimidazolo moiety.


Randomized, double-blind, placebo-controlled study of the effects of raloxifene and conjugated equine estrogen on plasma homocysteine levels in healthy postmenopausal women.
To investigate the long-term effects of raloxifene on fasting plasma homocysteine levels in postmenopausal women compared with conjugated equine estrogen (CEE). Randomized, double-blind, placebo-controlled study. Outpatient department of a university hospital. Fifty-two hysterectomized, healthy postmenopausal women. Oral raloxifene in two dosages (60 mg/d [n=13] and 150 mg/d [n=13]), oral CEE (0.625 mg/d [n=13], and placebo (n=13) were given for 24 months. Fasting plasma homocysteine concentrations. Plasma homocysteine levels were not altered in the placebo group. After 12 months, a significant reduction versus baseline in the mean plasma homocysteine level (-16%) was found only in the raloxifene 150-mg group. The mean change in plasma homocysteine levels within this group also was significantly different from the changes versus baseline found in the placebo group (+2%) and the raloxifene 60-mg group (-2%), but not different from those found in the CEE group (-8%). After 24 months, plasma homocysteine levels were decreased significantly in the raloxifene 150-mg and CEE groups compared with both baseline (-13% and -10%, respectively) and placebo values (-15% and -11%, respectively). No significant change in plasma homocysteine levels was observed in the raloxifene 60-mg group. Raloxifene has a favorable, dose-related effect on plasma homocysteine levels in postmenopausal women.


How Could These Mini Saccular Aneurysms of Ascending Aorta Be Classified?
The wall of a true aneurysm is composed of all histologic layers of the aorta. A false aneurysm represents a small, contained rupture of aorta followed by bulging of the corresponding area that is usually sustained by a fibrous peel. Aortic dissection is defined as a separation of the lamellae of the aortic wall. Herein, we describe an unusual clinical presentation of aortic dissection in a 37-year-old male patient that presented severe aortic regurgitation and unusual bulges with linear intimal fissures in ascending aorta underwent mechanical aortic valve replacement and interposition of tubular vascular graft in ascending aorta.


The contribution of MRI to the diagnosis of diffuse meningeal lesions.
We analysed MRI findings in patients in whom a diffuse abnormality of the meninges was revealed by MRI. We looked at T1 and T2-weighted spin-echo or fast spin-echo images and contrast-enhanced T1-weighted images. There were 15 patients with abnormalities on MRI, clinically suspected in ten. Four had meningoencephalitis, one meningeal and subcortical sarcoidosis nodules, four meningeal malignancies--one disseminated oligodendroglioma, one with meningeal infiltration around an adenocarcinoma, three meningeal infiltration by a haematological malignancy, and one a chronic subdural haematoma without a history of injury. We excluded patients with primary meningeal tumours and typical injury-related meningeal bleeding. The relatively small number of patients is due to both the infrequency of diffuse meningeal disease and to the low frequency of suspected meningeal pathology as an indication for MRI. The latter's diagnostic contribution is greatest in infectious disease and neoplastic infiltration, and less obvious in haematological malignancies. Contrast-enhanced T1-weighted images are most useful.


The influence of ultrasound on embryonic development.
The chick embryo provides a convenient experimental subject to assess the effects of ultrasound on a developing biological system. Insonating the whole embryo prior to organogenesis with spatial peak temporal average intensities of 4.5 W/cm2 pulsed, and 100 W/cm2 continuous wave focused, produced no lethal effect or observable developmental changes during three days following the treatment. Any bio-effect which may have resulted from the interaction of ultrasound at the cellular level was not severe enough to bring about irreversible gross physical abnormalities in the developing embryo. The effects of standing waves or other extraneous physical factors were considered to be greatly reduced by irradiating embryos "in ovo" rather than in glass dishes following explantation.


Therapeutic PCL scaffold for reparation of resected osteosarcoma defect.
Osteosarcomas are highly malignant tumors, which develop rapid growth and local infiltration, inducing metastases that spread primarily in the lung. Treatment of these tumors is mainly based on pre- and post-operative chemotherapy and surgery of the primary tumor. Surgical resection though, generates bone defects. Reparation of these weaknesses presents formidable challenges to orthopedic surgery. Medicine regenerative grafts that act as both tumor therapy with constant local drug delivery and tissue regeneration may provide a new prospect to address this need. These implants can provide sustained drug release at the cancer area, decreasing systemic second effects such as inflammation, and a filling of the resected tissues with regenerative biomaterials. In this study microporous poly-ε-caprolactone (PCL) scaffolds have been developed for sustained local release of anti-inflammatory drug dexamethasone (DXM), used as drug model, in cancer medicine regenerative field. The microporous PCL matrix of the scaffolds supported the attachment, proliferation and osteogenic differentiation of osteoblast-like cells, while the polyelectrolyte multilayers, anchored to the inner pore surfaces, sustained locally DXM release. These microporous scaffolds demonstrate the ability to deliver DXM as a localized tumor therapy and to promote proliferation and differentiation of osteoblast-like cells in vitro.


Effects of surface charge on denaturation of bovine carbonic anhydrase.
This work compares the denaturation of two proteins-bovine carbonic anhydrase II (BCA) and its derivative with all lysine groups acetylated (BCA-Ac(18))-by urea, guanidinium chloride (GuHCl), heat, and sodium dodecyl sulfate (SDS). It demonstrates that increasing the net negative charge of the protein by acetylation of lysines reduces its stability to urea, GuHCl, and heat, but increases its kinetic stability (its thermodynamic stability cannot be measured) towards denaturation by SDS. Increasing the ionic strength of the buffer improves the stability of BCA-Ac(18) to urea and heat, but still leaves it less stable than unacetylated BCA to those denaturants. In urea, the large change in electrostatic interactions not only modifies the free energy of denaturation, but also introduces a stable intermediate into the unfolding pathway. This work shows that modifications of charges on the surfaces of proteins can have a large effect--positive or negative, depending on the denaturant--on the stability of the proteins despite the exposure of these charges to high dielectric solvent and buffer ions.


Tissue plasminogen activator concentration dependence of 120 kHz ultrasound-enhanced thrombolysis.
It has been known for some time that the application of ultrasound can enhance the efficacy of thrombolytic medications such as recombinant tissue plasminogen activator (rt-PA). Potential clinical applications of this ultrasound-enhanced thrombolysis (UET) include the treatment of myocardial infarction, acute ischemic stroke, deep venous thrombosis and other thrombotic disorders. It may be possible to reduce the dose of rt-PA while maintaining lytic efficacy; however there is little data on the rt-PA concentration dependence of UET. In this work, the rt-PA concentration dependence of clot lysis resulting from 120 kHz UET exposure was measured in an in vitro human clot model. Clots were exposed to rt-PA for 30 min, with (UET treated) or without 120 kHz ultrasound (rt-PA treated) at 37 degrees C, and the clot width measured as a function of time. The rt-PA concentration ranged from 0-10 microg/mL. The initial lytic rate for the UET-treated group was greater than that of the rt-PA group at almost all rt-PA concentrations, and exhibited a maximum over concentration values of 1-3 microg/mL.


[Anti-drug antibodies, auto-antibodies and biotherapy in psoriasis].
The approval of substantial numbers of targeted biologic therapies (e.g., monoclonal antibodies, fusion proteins) for treatment of inflammatory diseases has positioned these drugs as important to fight chronic disorders such as psoriasis, rheumatoid arthritis and Crohn's disease. One of the concerns raised with the administration of biologic therapies is that because most of them are immunogenic glycoproteins they induce undesirable immune response leading to the generation of specific anti-drug antibodies (ADA). The development of "self" derived protein therapeutics (comprised of human germline sequence), such as recombinant "human" antibodies, helped to reduce the production of ADA but did not avoid all immunogenicity. Reduced efficacy and safety issues such as anaphylaxis or vasculitis accompany the development of ADA. In addition to immune reactions directed against the biologic therapies as a whole, some of them such as anti-TNFα are able to induce auto-immune response, notably antinuclear antibody (ANA). ANA development was associated with induced lupus and in psoriasis it was suggested that it may act as a marker of treatment failure to anti-TNFα. With a focus on psoriasis, this paper makes a current point on the consequences and challenges of the development of anti-drug antibodies and auto-immunity in patients who receive biologic therapies.


Efficacy of a novel oral formulation of sarolaner (Simparica™) against five common tick species infesting dogs in the United States.
The efficacy of a single oral treatment with sarolaner (Simparica™, Zoetis), a novel isoxazoline compound, was evaluated against five tick species known to infest dogs in the United States. A total of 10 laboratory studies, two against each species, were conducted using adult purpose-bred mongrels or Beagle dogs. In each study, 16 dogs were randomly allocated to one of two treatment groups based on pre-treatment host-suitability tick counts. Dogs were infested with approximately 50 unfed adult Amblyomma americanum, Amblyomma maculatum, Dermacentor variabilis, Ixodes scapularis or Rhipicephalus sanguineus ticks on Days -2, 5, 12, 19, 26 and 33. On Day 0, dogs were treated with a placebo or a sarolaner tablet providing a minimum dose of 2 mg/kg. Tick counts were conducted 48h after treatment and after each subsequent weekly re-infestation. There were no treatment-related adverse reactions during any of the studies. Dogs in the placebo-treated group maintained tick infestations throughout the studies. Geometric mean live tick counts were significantly lower (P≤0.0001) in the sarolaner-treated group compared to the tick counts in the placebo group at all timepoints. Treatment with sarolaner resulted in ≥99.6% efficacy against existing infestations of all five tick species within 48h. The efficacy against weekly post-treatment re-infestations of all tick species was ≥96.9% for at least 35 days after treatment. Thus, a single dose of sarolaner administered orally at the minimum dosage of 2mg/kg, resulted in excellent efficacy within 48h against existing tick infestations, and against weekly re-infestations for 35 days after treatment. These studies confirmed that administration of the minimum dose of sarolaner will provide rapid treatment of existing infestations and give at least one month of control against re-infestation by the common tick species affecting dogs in the US.


Telephone counseling and attendance in a national mammography-screening program a randomized controlled trial.
In Germany, a mammography-screening program (MSP) was implemented on a national level. It complies with all criteria of the European guidelines for quality assurance in screening mammography; however, the attendance rate is 54%, falling short of the target attendance rate of 70%. The aim of this study was to investigate whether additional telephone counseling improves attendance among nonresponders and the level of satisfaction with telephone counseling. In a prospective RCT, women identified as nonresponders in the MSP were randomized to a control group that received written reminders or to an intervention group that additionally received telephone counseling. In a follow-up, a subset of the intervention group was contacted by telephone regarding their satisfaction with telephone counseling. In 2008, a total of 5477 women aged 50-69 years who were eligible for the German MSP but had not participated up to 6 weeks after the first invitation were included in the study. Individual telephone counseling consisted of scripted calls from a trained counselor who provided information on MSP and answered the woman's questions. Report of mammography use provided by the screening unit 3 months after the reminder was sent. Analysis was conducted in 2009. Comparison of screening attendance revealed a significantly higher attendance rate in the intervention group compared with controls (29.7% vs 26.1%, p=0.0035). When only women for whom telephone numbers were available were analyzed, attendance was even better (35.5% vs 29.7%, p=0.0004). In the follow-up, 278 of 404 women were actually surveyed. Of those, 33% stated that telephone counseling had influenced their decision, 56% stated that they had undergone screening mammography, and 77% agreed that personal telephone counseling should be used routinely to encourage nonresponders to go for screening. Individual telephone counseling for nonresponders to a national program for breast cancer screening was well accepted by participants and effective. This study is registered at the Australian New Zealand Clinical Trials Registry ACTRN12611000645954.


Optimal criteria for the diagnosis of coronary artery disease by dobutamine stress echocardiography.
The diagnosis of coronary artery disease (CAD) on the basis of inducible ischemia in > or = 2, rather than 1, segments was suggested to improve specificity of dobutamine stress echocardiography (DSE). However, the impact of using these criteria on the sensitivity and accuracy of DSE was not studied. We studied the accuracy of DSE (up to 40 microg/kg/min) for the diagnosis of CAD in 290 patients with suspected myocardial ischemia using the criteria of > or = 1 and > or = 2 ischemic segments. Ischemia was defined as new or worsening wall motion abnormalities using a 16-segment model. Among the 85 patients without previous myocardial infarction, significant CAD was detected in 51 (60%). Sensitivity, specificity, and accuracy of DSE using > or = 1 ischemic segment were 73%, 85%, and 78%, respectively. Those using > or = 2 segments were 67%, 94%, and 78%, respectively (p = NS). Regional specificity improved by using > or = 2 segments (91% vs 96%, p <0.05) at the expense of an equivalent reduction in regional sensitivity (60% vs 44%, p <0.05), whereas the regional accuracy was similar (80% vs 79% for > or = 1 and > or = 2 segments, respectively). In the 205 patients with previous myocardial infarction, the criterion of ischemia in > or = 1 segment had a higher sensitivity and accuracy for overall diagnosis of CAD (75% vs 64%, p <0.05; 77% vs 68%, p <0.05), infarct-related CAD (64% vs 47%, p <0.005; 70% vs 57%, p <0.01), and remote CAD (74% vs 57%, p <0.005; 78% vs 69%, p <0.05) than the criterion of > or = 2 segments, respectively. It is concluded that in patients without previous myocardial infarction, the use of > or = 2 ischemic segments by DSE for the diagnosis of CAD does not improve the accuracy of DSE compared with the criterion of > or = 1 ischemic segment. Conversely, in patients with previous infarction the use of > or = 2 segments reduces the overall and regional sensitivity and accuracy without a significant improvement in specificity.


The impact of disease-related immobilization on thigh muscle mass and strength in older hospitalized patients.
We assessed the quantitative changes in muscle mass and strength during 2 weeks of hospitalization in immobile and mobile acutely ill hospitalized older adults. Forty-one patients (82.4 ± 6.6 years, 73.0% females) participated in this prospective longitudinal observational study. Mobility status was defined according to walking ability as described in the Barthel-Index. Functional status, including handgrip strength and isometric knee-extension strength, and mid-thigh magnetic resonance imaging (MRI) measurements of cross-sectional area (CSA) were conducted on admission and at discharge. Twenty-two participants (54%) were immobile and 19 (46%) mobile. In all, 54.0 and 12.0% were at risk of malnutrition and malnourished, respectively. The median time between baseline and follow-up for MRI scans were 13 days in mobile and immobile participants (P = 0.072). Mid-thigh muscle and subcutaneous fat CSA significantly decreased by 3.9cm2 (5.0%, P = 0.002) and 5.3cm2 (5.7%, P = 0.036) during hospitalization whereas intermuscular fat remained unchanged in immobile subjects. No significant changes were observed in mobile patients. In a regression analysis, mobility was the major independent risk factor for changes in mid-thigh muscle CSA as a percentage of initial muscle area (P = 0.022) whereas other variables such as age (P = 0.584), BMI (P = 0.879), nutritional status (P = 0.835) and inflammation (P = 0.291) were not associated with muscle mass changes. There was a significant decrease in isometric knee extension strength (P = 0.002) and no change in handgrip strength (P = 0.167) in immobile patients whereas both parameters increased significantly over time in mobile patients (P = 0.048 and P = 0.012, respectively). Two weeks of disease-related immobilization result in a significant loss of thigh muscle mass and muscle strength in older patients with impaired mobility. Concomitantly, there was a significant reduction of subcutaneous adipose tissue in immobile older hospitalized patients whereas no changes were observed in intermuscular fat among these patients. These data highlight the importance of mobility support in maintaining muscle mass and function in older hospitalized patients.


Empathy and Theory of Mind in Ultra-High Risk for Psychosis: Relations With Schizotypy and Executive Function.
While recent studies have found deficits in theory of mind (ToM) skills in individuals at ultra-high risk (UHR) for psychosis, empathic tendencies in these subjects remain unclear. The presence of high schizotypy and compromised executive functions, which are found in UHR individuals, would affect ToM skills and empathic tendencies. We investigated the ToM skills and empathic tendencies of UHR individuals and examined their relationship with schizotypy and executive function. This study included 28 UHR individuals and 28 age- and sex-matched healthy controls. All participants completed a self-reported empathic scale (Interpersonal Reactivity Index) and the Wisconsin Schizotypy Scales. Additionally, the ToM Picture Stories Task and Wisconsin Card Sorting Test were conducted. UHR individuals showed a trend toward lower self-reported empathic tendencies; however, there were no differences in ToM skills between the two groups. Of the four subscales of the IRI, only empathic concern showed a significant difference between the two groups. Empathic concern was inversely associated with negative schizotypy. Our findings suggest that UHR individuals show relatively preserved cognitive empathy but compromised emotional empathy. Furthermore, in UHR individuals, the empathic concern subscale of the IRI was associated with negative schizotypy, while ToM skills were related to positive schizotypy and executive function.


Effect of selected dimethylaminochalcones on some mitochondrial activities.
Chalcones and their synthetic cyclic analogues have been shown to possess a full scale of biological activities in a variety of experimental systems. They were assessed to be mostly effective in defense against free radicals in the organism, but several compounds exhibited cytotoxic pro-oxidant activities. The respiratory response and antioxidant status in mitochondria were investigated upon addition of 4'-dimethylaminochalcone (1a) and its cyclic analogues, (E)-2-(4'-((CH3)2 N)-benzylidene)-1-indanone (1b), -1-tetralone (1c), and -1-benzosuberone (1d). Selected structures were able to change the respiratory response of mitochondria and showed an ability to modify mitochondrial metabolic and redox efficiency, though they did not indicate redox reactivity towards glutathione in adduct-free incubations. The results of the study indicate that -chalcone and -tetralone derivatives cause suppression of reactive oxygen species affecting mitochondrial respiration by mild uncoupling. In addition, (E)-2-(4'-((CH3)2 N)-indanone (1b), and to a greater extent, -benzosuberone (1d), showed pro-oxidant effects, which partially explain their cytotoxicity.


Recent Advances on Metalloporphyrin-Based Materials for Visible-Light-Driven CO2 Reduction.
Photocatalytic CO2 reduction is a promising technology to mitigate environmental issue and the energy crisis. The four nitrogen atoms in the porphyrin ring can incorporate transition metals to form stable active sites for CO2 activation and photoreduction. Nevertheless, the photocatalytic efficiency of metalloporphyrins is still low due to the insufficient photoelectron injection to drive CO2 photoreduction upon visible light irradiation. To address this issue, considerable efforts have been made to introduce photosensitizers for constructing homogeneous or heterogeneous metalloporphyrin-based photocatalytic systems. In this Review, recent advances of metalloporphyrin-based materials for visible-light-driven CO2 reduction were summarized. The methods for the modulation of photosensitizing process at molecular level were presented for the promotion of photocatalytic performance. The mechanism of CO2 activation and photocatalytic conversion was illustrated. Better insight into the structure-activity relationship provides guidance to the design of metalloporphyrin-related photocatalytic systems.


Amount of airflow required for olfactory perception in laryngectomees: a prospective interventional study.
To determine minimum airflow rate required for olfactory stimulation in successfully rehabilitated laryngectomised patients after learning the polite yawning technique (PYT) and to confirm the hypothesis that sense of smell is rehabilitated once the nasal airflow is re-established. Prospective open interventional trial. Tertiary academic hospital. The study population comprised 100 laryngectomised patients. The control group consisted of 100 non-laryngectomised patients of similar age and sex. Rhinomanometry was used to measure air flow in the right and left nostrils, respectively, while the Smell Diskettes Olfaction test (SDOT) was used to test each individual's sense of smell. The primary endpoint was increasing the airflow, while the secondary endpoint was improvement in the Smell Diskettes Olfaction test score after learning the polite yawning technique. The difference in the Smell Diskettes Olfaction test results before and after introducing the polite yawning technique was statistically significant (F = 53.077; P < 0.001). The number of accurately identified odours increased with each measurement. There was a significant difference among rhinomanometric measurements of airflow through the right (F = 65.002; P < 0.001) and left nostrils (F = 75.465; P < 0.001). Nasal airflow improved with each measurement. The minimum airflow required for olfactory stimulation in successfully rehabilitated patients was approximately 60 cm(3) /s. The control group had considerably better airflow in both nostrils than the laryngectomised group. The difference between the total number of rehabilitated (normosmic) patients (48%) in the laringectomised group and normosmic participants (56%) in the control group (z = 1.132; P = 0.129) was not statistically significant. The number of odours identified by laryngectomised patients increased with the volume of nasal airflow. The number of patients with rehabilitated olfactory function approximated the percentage of normosmic individuals in the non-laryngectomised population. These findings confirm the hypothesis that sense of smell is rehabilitated once the nasal airflow is re-established.


Uncharted Waters: Challenges in the Era of Biologic Therapies for Nasal Polyposis.
Several new, promising, targeted therapies for nasal polyposis are being tested in large-scale clinical trials. These agents target pathways thought to be involved in the disease, including IgE, IL-5, IL-4/IL-13, and others. Designing these trials poses significant challenges: who and when to enroll is not completely clear, optimal dosing is not known, outcome measures are insufficiently robust, there are no validated biomarkers, trial regimens may not comport with how clinicians might use these drugs once approved, and cost-benefit ratios have not been assessed. Thus, there is a need to consider such questions, as trials of these novel treatments continue and these biologics become available. Despite these uncertainties about trial design, there remains a great deal of excitement in the field as we approach the dawn of a new era of therapeutic options for nasal polyposis. In this rostrum, we enumerate these issues and call for a conference that will allow stakeholders in the field to confront them as we enter this new era of opportunity to advance the treatment of nasal polyposis.


[Experimental research on spine fusion induced by tissue engineered bone].
To construct a reasonable substitute for the autograft bone in vitro and transplant it back into the rabbit models to induce the spine fusion. The bone marrow stem cell from the seven New Zealand rabbits were cultured. Recombinant human bone morphogenetic protein-4 (rhBMP-4) that has been proved to be bioactive was obtained by the way of genetic engineering. Using the vacuum freezing machine to mix a certain quantity of rhBMP-4 into type I collagen to form a new kind of carrier. Animal model of spine facet process fusion was used. Bone marrow stem cells combined with rhBMP-4 and I type collagen were implanted between the facet process to induce the spine fusion. type I collagen and bone marrow stem cell was used in the controlled group. New bone formation was obvious in the test group. The facet joint was fused very well in this side. No bone formation was present on the other side. The new composite: bone marrow stem cells, rhBMP-4 and type I collagen was an ideal kind of substitute for the autograft bone.


Amitriptyline and nortriptyline serum determination by micellar liquid chromatography.
Amitriptyline and nortriptyline are tricyclic antidepressants which act by enhancing the actions of norepinephrine and serotonin caused by blocking the re-uptake of various neurotransmitters at the neuronal membrane. A micellar liquid chromatographic procedure was developed to determine these drugs in serum samples for use in clinical monitoring. The chromatographic determination of these highly hydrophobic substances was carried out using a 0.15 M SDS-6% (v/v) pentanol buffered at pH 7, in a C18 column, and electrochemical detection at 650 mV. The flow-rate was 1.5 mL/min. The analysis time was 14 min. The limits of detection (ng/mL) in serum were 0.25 and 0.31 for amitriptyline and nortriptyline, respectively. Repeatability and intermediate precision were evaluated at three different concentrations in serum samples. Untreated serum samples were injected directly into the HPLC system after filtration, leading to be a simple procedure that can be applied in routine analyses for Therapeutic Drug Monitoring.


The mycobacterial cell envelope - a moving target.
Mycobacterium tuberculosis, the leading cause of death due to infection, has a dynamic and immunomodulatory cell envelope. The cell envelope structurally and functionally varies across the length of the cell and during the infection process. This variability allows the bacterium to manipulate the human immune system, tolerate antibiotic treatment and adapt to the variable host environment. Much of what we know about the mycobacterial cell envelope has been gleaned from model actinobacterial species, or model conditions such as growth in vitro, in macrophages and in the mouse. In this Review, we combine data from different experimental systems to build a model of the dynamics of the mycobacterial cell envelope across space and time. We describe the regulatory pathways that control metabolism of the cell wall and surface lipids in M. tuberculosis during growth and stasis, and speculate about how this regulation might affect antibiotic susceptibility and interactions with the immune system.


[Prevention and management of complications with laparoscopy cholecystectomy].
To prevent the complication of laparoscopy cholecystectomy (LC). This text analysed 1,200 cases retrospectively with LC from Feb. 1993 to May. 1998. Three cases with biliferous duct injury, two cases with bile leakage from the stamp of cystic duct, two cases with gallbladder artery injury. Strict preoperative examination and standard operative procedure can reduce markedly complication of LC.


Antimicrobial activity in Asterceae: The selected genera characterization and against multidrug resistance bacteria.
Plants from the Asteraceae family are widely used as ethno medicines to treatment parasitic, malaria, hematemesis, pruritus, pyretic, anthelmintic, wound healing. The aim of this review is to provide an overview of Asteraceae plants antimicrobial activity. The most relevant results from the published studies are summarized and discussed. The species in genus of Artemisia, Echinacea, Centaurea, Baccharis, and Calendula showed antimicrobial activity. Most of these species are usually used as ethno medicines to treat infection, inflammation, and parasitics. The effective part or component for antimicrobial was essential oil and crude extract, and essential oil attracted more attention. It was also reported that nanoparticles coated with crude extract were effective against multidrug resistant bacteria. For multidrug resistant bacteria study, the species in Armtemisia were the most investigated, and Staphylococcus aureus and Escherichia coli were the most studied multidrug resistant strains. The antimicrobial activity was evaluated mainly based on the results of minimum inhibitory concentration (MIC). Few reports have been reported on minimum bactericide concentration (MBC) and its antibacterial mechanisms. According to the reported study results, some plants in Asteraceae have the potential to be developed as bacteriostatic agents and against multidrug resistant bacteria. However, most studies are still in vitro, further clinical and applied studies are needed.


Characterization of the Tuber borchii nitrate reductase gene and its role in ectomycorrhizae.
The nitrate assimilation pathway represents a useful model system in which to study the contribution of a mycorrhizal fungus to the nitrogen nutrition of its host plant. In the present work we cloned and characterized the nitrate reductase gene (tbnr1) from Tuber borchii. The coding region of tbnr1 is 2,787 nt in length, and it encodes a protein of 929 amino acids. Biochemical and Northern-blot analyses revealed that nitrate assimilation in T. borchii is an inducible system that responds mainly to nitrate. Furthermore, we cloned a nitrate reductase cDNA (tpnr1) from Tilia platyphyllos to set up a quantitative real-time PCR assay that would allow us to determine the fungal contribution to nitrate assimilation in ectomycorrhizal tissue. Using this approach we demonstrated that the level of tbnr1 expression in ectomycorhizae is eight times higher than in free-living mycelia, whereas tpnr1 transcription was found to be down-regulated after the establishment of the symbiosis. Enzymatic assays showed that NADPH-dependent nitrite formation markedly increases in ectomycorrhizae. These findings imply that the fungal partner plays a fundamental role in nitrate assimilation by ectomycorrhizae. Amino acid determination by HPLC revealed higher levels of glutamate, glutamine and asparagine in symbiotic tissues compared with mycelial controls, thus suggesting that these amino acids may represent the compounds that serve to transfer nitrogen to the host plant.


Does N2 fixation meet the nitrogen requirements of heterocystous blue-green algae in shallow eutrophic lakes?
Phytoplankton net carbon uptake and nitrogen fixation were studied in two shallow, eutrophic lakes in South Sweden. Ranges of diurnal net carbon uptake were estimated by subtracting 24-h respiration rates corresponding to 5-20% of P max, respectively, from daytime carbon uptake values. total nitrogen requirement of the phytoplankton assemblage was determined from the diurnal net carbon uptake, assuming a phytoplankton C:N ratio of 9.5:1. Nitrogen supplied by nitrogen fixation only occasionally corresponded to the demands of the total phytoplankton assemblage. When heterocystous algae made up a substantial proportion (≥10%) of the total phytoplankton biomass, nitrogen fixation could meet the requirements of heterocystous blue-green algae on c. 50% of the sampling occasions. Nitrogen deficiencies in heterocystous algae were most probably balanced by the simultaneous or sequential assimilation of dissolved inorganic nitrogen. It was concluded that uptake of ammonium or nitrate, regenerated from lake seston and sediment, is the main process by which growth of phytoplankton is maintained during summer in the lake ecosystems studied.


Molecular and pharmacological characterization of genes encoding urotensin-II peptides and their cognate G-protein-coupled receptors from the mouse and monkey.
Urotensin-II (U-II) and its receptor (UT) represent novel therapeutic targets for management of a variety of cardiovascular diseases. To test such hypothesis, it will be necessary to develop experimental animal models for the manipulation of U-II/UT receptor system. The goal of this study was to clone mouse and primate preproU-II and UT for pharmacological profiling. Monkey and mouse preproU-II genes were identified to encode 123 and 125 amino acids. Monkey and mouse UT receptors were 389, and 386 amino acids, respectively. Genomic organization of mouse genes showed that the preproU-II has four exons, while the UT receptor has one exon. Although initially viewed by many exclusively as cardiovascular targets, the present study demonstrates expression of mouse and monkey U-II/UT receptor mRNA in extra-vascular tissue including lung, pancreas, skeletal muscle, kidney and liver. Ligand binding studies showed that [125I]h U-II bound to a single sites to the cloned receptors in a saturable/high affinity manner (Kd 654+/-154 and 214+/-65 pM and Bmax of 1011+/-125 and 497+/-68 fmol mg-1 for mouse and monkey UT receptors, respectively). Competition binding analysis demonstrated equipotent, high affinity binding of numerous mammalian, amphibian and piscine U-II isopeptides to these receptors (Ki=0.8 - 3 nM). Fluorescein isothiocyanate (FITC) labelled U-II, bound specifically to HEK-293 cells expressing mouse or monkey UT receptor, confirming cell surface expression of recombinant UT receptor. Exposure of these cells to human U-II resulted in an increase in intracellular [Ca2+] concentrations (EC50 3.2+/-0.8 and 1.1+/-0.3 nM for mouse and monkey UT receptors, respectively) and inositol phosphate (Ip) formation (EC50 7.2+/-1.8 and 0.9+/-0.2 nM for mouse and monkey UT receptors, respectively) consistent with the primary signalling pathway for UT receptor involving phospholipase C activation.


[Expression of vascular endothelial growth factor C in laryngeal squamous carcinoma and its role in lymphatic metastasis].
To study the expression of vascular endothelial growth factor C (VEGF-C) in laryngeal squamous carcinoma (LSC) and its relationship to lymph node metastasis. Paraffin-embedded specimens of primary laryngeal carcinomas from 60 patients subjected to operation were studied. VEGF-C expression was investigated using immunohistochemical staining. Positive immunohistochemical staining for VEGF-C protein was observed in some laryngeal squamous carcinoma cells (45/60), the VEGF-C antigen was identified in the cytoplasm of cancer cells. Furthermore, the expression of VEGF-C in laryngeal squamous carcinoma cells was significantly higher in lymph-node-positive groups than in node-negative groups, and the expression was significantly higher in poorly differentiated LSC tissues than in moderately and well differentiated LSC tissues. The above findings indicate a close correlation between VEGF-C expression and lymph node metastasis in laryngeal squamous carcinoma.


Actin-mediated retraction of the larval epidermis during metamorphosis of the sand dollar, Dendraster excentricus.
During the first 15 to 20 min of metamorphosis the larval arms are retracted and resorbed into the aboral surface of the juvenile. Arms excised from metamorphosing larvae will undergo a sequence of contraction and histolysis that is identical to that occurring in intact larvae. Prior to and during metamorphosis, epidermal cells contain bundles of 5 to 7-nm microfilaments in arrays radiating apically from the base of the cells. Sparse microfilaments also occur near the plasmalemma of epidermal cells and some mesenchymal cells in larvae fixed during metamorphosis. Contraction of excised arms is reversibly inhibited by treatment with cytochalasin B, and microfilaments bind myosin sub-fragment-1. Indirect immunofluorescence of larval arms using an antibody against chicken-muscle actin and staining with the F-actin specific probe, NDB phallacidin indicate that the arms contain actin distributed in a manner consistent with ultrastructural findings. It is proposed that retraction of the larval arms during metamorphosis is produced by an actin-mediated change in shape of the epidermal cells.


Double pulse laser induced breakdown spectroscopy: A potential tool for the analysis of contaminants and macro/micronutrients in organic mineral fertilizers.
Organic fertilizers are obtained from waste of plant or animal origin. One of the advantages of organic fertilizers is that, from the composting, it recycles waste-organic of urban and agriculture origin, whose disposal would cause environmental impacts. Fast and accurate analysis of both major and minor/trace elements contained in organic mineral and inorganic fertilizers of new generation have promoted the application of modern analytical techniques. In particular, laser induced breakdown spectroscopy (LIBS) is showing to be a very promising, quick and practical technique to detect and measure contaminants and nutrients in fertilizers. Although, this technique presents some limitations, such as a low sensitivity, if compared to other spectroscopic techniques, the use of double pulse (DP) LIBS is an alternative to the conventional LIBS in single pulse (SP). The macronutrients (Ca, Mg, K, P), micronutrients (Cu, Fe, Na, Mn, Zn) and contaminant (Cr) in fertilizer using LIBS in SP and DP configurations were evaluated. A comparative study for both configurations was performed using optimized key parameters for improving LIBS performance. The limit of detection (LOD) values obtained by DP LIBS increased up to seven times as compared to SP LIBS. In general, the marked improvement obtained when using DP system in the simultaneous LIBS quantitative determination for fertilizers analysis could be ascribed to the larger ablated mass of the sample. The results presented in this study show the promising potential of the DP LIBS technique for a qualitative analysis in fertilizers, without requiring sample preparation with chemical reagents.


Study and analysis of different segmentation methods for brain tumor MRI application.
Medical Resonance Imaging (MRI) is one of the preferred imaging methods for brain tumor diagnosis and getting detailed information on tumor type, location, size, identification, and detection. Segmentation divides an image into multiple segments and describes the separation of the suspicious region from pre-processed MRI images to make the simpler image that is more meaningful and easier to examine. There are many segmentation methods, embedded with detection devices, and the response of each method is different. The study article focuses on comparing the performance of several image segmentation algorithms for brain tumor diagnosis, such as Otsu's, watershed, level set, K-means, HAAR Discrete Wavelet Transform (DWT), and Convolutional Neural Network (CNN). All of the techniques are simulated in MATLAB using online images from the Brain Tumor Image Segmentation Benchmark (BRATS) dataset-2018. The performance of these methods is analyzed based on response time and measures such as recall, precision, F-measures, and accuracy. The measured accuracy of Otsu's, watershed, level set, K-means, DWT, and CNN methods is 71.42%, 78.26%, 80.45%, 84.34%, 86.95%, and 91.39 respectively. The response time of CNN is 2.519 s in the MATLAB simulation environment for the designed algorithm. The novelty of the work is that CNN has been proven the best algorithm in comparison to all other methods for brain tumor image segmentation. The simulated and estimated parameters provide the direction to researchers to choose the specific algorithm for embedded hardware solutions and develop the optimal machine-learning models, as the industries are looking for the optimal solutions of CNN and deep learning-based hardware models for the brain tumor.


Aminoalkyl functionalization of dextran for coupling of bioactive molecules and nanostructure formation.
Aminopropyl dextrans and mixed aminopropyl cyanoethyl dextrans were prepared from cyanoethyl precursors by full or partial reduction with CoCl2/NaBH4. Coupling of various aldehydes to the glucan backbone by reductive amination was accomplished with 4-hydroxy-3-methoxybenzaldehyde (vanillin), 3,5-di-tert-butyl-4-hydroxy-benzaldehyde (BHT-CHO), maltose and maltotriose, and picoline borane as reducing agent. Successful coupling of these representatives for aroma compounds, antioxidants and sugar side-chains were verified by ESI-MS after hydrolysis and by 1D and 2D NMR spectroscopy. Degree of conversion (molar ratio of coupled aldehydes) was estimated from (1)H NMR spectra. Formation of secondary and tertiary amines was detectd by ESI-MS. Applying a solvent exchange process, new nanoparticles based on these modified dextrans were prepared with and without addition of iron oxide nanoparticles.


Septic shock caused by a carbon dioxide-dependent and extended spectrum β-lactamase-producing Proteus mirabilis small colony variant in a long-term bedridden patient.
Here, we report a 60-year-old chronically bedridden man with cerebral palsy who had septic shock following a history of urinary tract infection with extended spectrum β-lactamase-producing and auxotrophic Proteus mirabilis detected on blood and urine cultures. This auxotroph formed small colonies only on the blood agar at 24 h in 5% CO2, but not in the conditions without CO2, and lacked motility and some biochemical activities. The five-year history of stones in the right renal pelvis suggests chronic urinary tract infection with P. mirabilis requiring a 28-day antibiotic treatment. This paper highlights that the CO2-dependent P. mirabilis small colony variant may cause sepsis, probably due to chronic infection in uroliths, which should warrant immediate identification.


Kidney transplant recipients with cutaneous squamous cell carcinoma have an increased risk of internal malignancy.
This study aimed to investigate whether the occurrence of cutaneous squamous cell carcinomas (SCCs) is associated with an increased risk of internal malignancies (IMs) in kidney transplant recipients (KTRs). In a cohort study, all patients receiving kidney transplantation in Leiden, the Netherlands, between 1966 and 2006 were followed up. All malignancies that had developed between 1966 and 2007 were recorded. Time-dependent Cox regression analyses were used to calculate the association between the development of cutaneous SCCs and IMs. The incidence of IMs in the KTRs after transplantation was also compared with the general Dutch population by calculating standardized morbidity ratios (SMRs) and was matched for age, sex, and time period in which the malignancy had occurred. Among 1,800 KTRs, 176 (9.8%) developed cutaneous SCCs and 142 (7.9%) developed IMs after transplantation. In patients with prior cutaneous SCCs, the adjusted risk to develop IMs was 3.0 (1.9; 4.7). In KTRs without cutaneous SCCs, the risk of IM compared with the general population was hardly increased. KTRs with cutaneous SCCs have an increased risk to develop IMs, and this information can be used to identify KTRs who are at an increased risk for IMs.


Brain organization: wiring economy works for the large and small.
The highest-resolution test to date of the wire minimization hypothesis has found that this principle works well for brain regions with a volume just over 400 μm(3). What is the wire minimization hypothesis, and why should anyone care about it?


Determination of the CYP1A-inducing potential of single substances, mixtures and extracts of samples in the micro-EROD assay with H4IIE cells.
This protocol describes a quantitative and robust 96-well-plate-reader-based assay for the measurement of ethoxyresorufin-O-deethylase (EROD) activity using the rat hepatoma cell line H4IIE. The assay can be used to determine the cytochrome P450 subfamily 1A (CYP1A)-inducing potential of single substances, as well as of mixtures and extracts of samples. It is based on the aryl hydrocarbon receptor (AhR)-mediated induction of cytochrome P450 enzymes (subfamily 1A) in cells after exposure to dioxins and dioxin-like compounds. One enzymatic reaction catalyzed by CYP1A is the deethylation of the exogenous substrate 7-ethoxyresorufin to the fluorescent product resorufin, which is measured as EROD activity in the assay. The CYP1A-inducing potential of a sample can be reliably quantified by comparing the EROD activity with the concentration-response curve of the standard substance 2,3,7,8-tetrachlorodibenzo-p-dioxin, which can be detected at concentrations down to the picogram per liter range. A researcher familiar with the procedure can process up to 160 samples with four wells each within 3 d. The series described uses four plates with three concentrations per sample, which can be easily scaled to accommodate different sample sizes.


Renal magnetic resonance angiography in the preoperative detection of supernumerary renal arteries in potential kidney donors.
Careful evaluation of the renovascular anatomy in potential living-related kidney donors is essential regarding the presence of accessory renal arteries. Conventional arteriography remains the standard of evidence for delineating the renal arterial supply. We assessed the utility of two-dimensional phase contrast (PC) magnetic resonance angiography (MRA) in the workup of potential living-related renal donors. Thirty-nine patients were examined with conventional arteriography and MRA, which was performed on a 1.5-T system using a two-dimensional PC technique in both coronal and axial planes (repetition time[TR]/echo time [TE] = 39/8.5 msec; flip 60 degrees; matrix 256 x 128; field of view, 28 cm2; 2 excitations; first-order gradient-moment nulling; 7-mm section with 2-mm overlap). The number of hilar and polar supernumerary renal arteries was determined. Hilar supernumerary arteries were classified as co-dominant if they were similar in size to the ipsilateral main renal artery. Conventional arteriography identified 78 dominant and 13 supernumerary (3 co-dominant, 10 accessory) renal arteries. Magnetic resonance angiography identified the proximal 35 mm of all 78 dominant and the 3 co-dominant renal arteries. Of the remaining ten (7 polar and 3 hilar) accessory vessels, only four were correctly identified with MRA. The high error rate (60%) suggests that two-dimensional PC MRA, as implemented, should not be used in the preoperative evaluation of potential renal donors.


Therapy Effect: Impact on Bone Marrow Morphology.
This article highlights the most common morphologic features identified in the bone marrow after chemotherapy for hematologic malignancies, growth-stimulating agents, and specific targeted therapies. The key is to be aware of these changes while reviewing post-therapeutic bone marrow biopsies and to not mistake reactive patterns for neoplastic processes. In addition, given the development and prevalent use of targeted therapy, such as tyrosine kinase inhibitors and immune modulators, knowledge of drug-specific morphologic changes is required for proper bone marrow interpretation and diagnosis.


Quantification of variation and the impact of biomass in targeted 16S rRNA gene sequencing studies.
Recent advances in sequencing technologies and bioinformatics tools have allowed for large-scale microbiome studies that are rapidly advancing medical research. However, small changes in technique or analysis can significantly alter the results and lead to conflicting findings. Quantifying the technical versus biological variation expected in targeted 16S rRNA gene sequencing studies and how this variation changes with input biomass is critical to guide meaningful interpretation of the current literature and plan future research. Data were compiled from 469 sequencing libraries across 19 separate targeted 16S rRNA gene sequencing runs over a 2.5-year time period. Following removal of contaminant sequences identified from negative controls, 244 samples retained sufficient reads for further analysis. Coefficients of variation for intra- and inter-assay variation from repeated measurements of a bacterial mock community ranged from 8.7 to 37.6% (intra) and 15.6 to 80.5% (inter) for all but one genus of bacteria whose relative abundance was greater than 1%. Intra- versus inter-assay Bray-Curtis pairwise distances for a single stool sample were 0.11 versus 0.31, whereas intra-assay variation from repeat stool samples from the same donor was greater at 0.38 (Wilcoxon p = 0.001). A dilution series of the bacterial mock community was used to assess the effect of input biomass on variability. Pairwise distances increased with more dilute samples, and estimates of relative abundance became unreliable below approximately 100 copies of the 16S rRNA gene per microliter. Using this data, we created a prediction model to estimate the expected variation in microbiome measurements for given input biomass and relative abundance values. Well-controlled microbiome studies are sufficiently robust to capture small biological effects and can achieve levels of variability consistent with clinical assays. Relative abundance is negatively associated with measures of variability and has a stronger effect on variability than does absolute biomass, suggesting that it is feasible to detect differences in bacterial populations in very low-biomass samples. Further, by quantifying the effect of biomass and relative abundance on compositional variability, we developed a tool for defining the expected variance in a given microbiome study.


Effect of early stellate ganglion block in cerebral vasospasm after aneurysmal subarachnoid hemorrhage (BLOCK-CVS): study protocol for a randomized controlled trial.
Stellate ganglion block has been reported to expand cerebral vessels and alleviate vasospasm after aneurysmal subarachnoid hemorrhage. However, the causal relationship between early stellate ganglion block and cerebral vasospasm prevention has not yet been established. The purpose of this study was to explore the effectiveness and safety of early stellate ganglion block as a preventive treatment for cerebral vasospasm and delayed cerebral ischemia. This is a single-center, prospective, randomized, controlled, blinded endpoint assessment superiority trial. A total of 228 patients will be randomized within 48 h of aneurysmal subarachnoid hemorrhage onset in a 1:1 ratio into two groups, one group receiving an additional e-SGB and the other group receiving only a camouflaging action before anesthesia induction in the operating room. The primary outcome is the incidence of symptomatic vasospasm within 14 days after aSAH. Further safety and efficacy parameters include the incidence of radiographic vasospasm, new cerebral infarction, postoperative delirium, and complications up to 90 days after surgery; postoperative cerebral hemodynamics; Mini-Mental State Examination score; modified Rankin scale score; and all-cause mortality up to 90 days after surgery. This is a randomized controlled trial to explore the effectiveness and safety of early stellate ganglion block as a preventive treatment to reduce cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage. If the results are positive, it may provide a new direction for the prevention and treatment of cerebral vasospasm and delayed cerebral ischemia. The study was registered on Clincaltrials.gov on December 13, 2020 (NCT04691271).


Biomarkers of selenium status in the Amazonian context: blood, urine and sequential hair segments.
Selenium (Se) is an essential element and deficit or excess of dietary Se is associated with health disorders. Relatively elevated Se levels have been reported in the Brazilian Amazon, where there are also important annual variations in the availability of different foods. The present study was conducted among six riparian communities of the Tapajós River to evaluate seasonal variations in blood and sequential hair cm Se concentrations, and to examine the relationships between Se in blood and hair, and blood and urine. Two cross-sectional studies were conducted, at the descending water (DWS, n=259) and the rising water (RWS, n=137) seasons, with repeated measures for a subgroup (n=112). Blood Se (B-Se), hair Se (H-Se) and urine Se (U-Se) were determined. Match-paired analyses were used for seasonal comparisons and the method of best fit was used to describe the relationships between biomarkers. B-Se levels presented a very large range (142-2447 microg/l) with no overall seasonal variation (median 284 and 292 microg/l, respectively). Sequential analysis of 13 cm hair strands showed significant variations over time: Se concentrations at the DWS were significantly lower compared with the rising water season (medians: 0.7 and 0.9 microg/g; ranges: 0.2-4.3 microg/g and 0.2-5.4 microg/g, respectively). At both seasons, the relationships between B-Se and H-Se were linear and highly significant (r(2)=67.9 and 63.6, respectively), while the relationship between B-Se and U-Se was best described by a sigmoid curve. Gender, age, education and smoking did not influence Se status or biomarker relationships. Variations in H-Se suggest that there may be seasonal availability of Se sources in local food. For populations presenting a large range and/or elevated Se exposure, sequential analyses of H-Se may provide a good reflection of variations in Se status.


Photophysical properties of popular fluorescent adenosine nucleotide analogs used in enzyme mechanism probing.
Fluorescent nucleotide analogs are widely used in mechanistic studies of nucleotide binding and utilizing proteins. We describe here an overview of the photophysical parameters of the most popular nucleotide analogs that have a fluorescent N-methylanthraniloyl-group attached at various positions of the nucleotide. Steady state absorption and fluorescence spectra of free chromophores depend on the type of modification (ribose, base or phosphate moiety) and the addition of proteins suggests that the labeled nucleotides also vary in sensitivity depending upon their local protein environment. Fluorescence lifetime measurements imply two to three lifetimes for each nucleotide with complex changes in dependence on solvent but more importantly also on the protein. The measured quantum yields quantify the increase in fluorescence for (C8)-MABA-ADP, MANT-ATP and (Pgamma)-MABA-ATP as 153%, 93% and 14% when bound to DnaK, ClpB and Trap1, respectively, compared to free in buffer solution.


[Endovascular treatment of traumatic lesions of thoracic aorta. Our experience].
To report our experience with endovascular repair of descending thoracic aorta in patients with acute and chronic traumatic lesions due to blunt chest trauma in last 4 years. From January 2002 to September 2006, 5 patients underwent endovascular repair of descending thoracic aorta for acute (3 cases) and chronic (2 cases) traumatic lesions due to road traffic accidents (motorcycle crash: 1; car crash: 4). Stent-graft placement was successful in all cases with good early outcomes and without late complications (follow up 3-48 months). Endovascular repair of descending thoracic aorta lesions, due to blunt chest trauma, is a feasible and valid alternative to conventional open operation.


Characterization of the chlorpropamide-alcohol-flush in patients with type 1 and type 2 diabetes.
The aim of the present paper was to evaluate the prevalence of the chlorpropamide-alcohol-flush (CPAF) in patients with type 2 and with type 1 diabetes. Ninety-seven patients with type 2 diabetes and 33 with type 1 diabetes drank 40 ml vermouth 12 h after placebo and again 12 h after 1 tablet of chlorpropamide (250 mg) or 12 h after the last of repeated administrations of chlorpropamide (250 mg b.i.d. for 2 days). Skin temperature was recorded in all patients by a thermocouple probe connected to the left cheek. In 47 patients serum concentrations of chlorpropamide and of its metabolite CBSU were also determined. The prevalence of CPAF was similar in type 1 and type 2 diabetes, was greater in women than in men, and was significantly greater after repeated administrations than after one single administration of chlorpropamide. The increase of skin temperature during a 30-min period was significantly higher in patients with CPAF than in patients without CPAF. Serum concentrations of chlorpropamide and of its metabolite CBSU were more elevated after 4 than after 1 tablet of chlorpropamide, but were not significantly different in patients with and without CPAF. These data indicate that both genetic factors and the amount of chlorpropamide used affect the appearance of CPAF. To assess the possible role of serotonin and of dopamine in the CPAF, some patients with CPAF were tested again after treatment with metergoline, an antiserotonin agent, or with bromocriptine, a dopamine-agonist. Neither drug influenced the CPAF, indicating that the two neurotransmitters are not involved in the CPAF.


In vivo temperature controlled ultrasound-mediated intracellular delivery of cell-impermeable compounds.
Many chemotherapeutic drugs are characterized by high systemic toxicity and/or suffer from limited bioavailability. Thermosensitive liposomes (TSLs) encapsulating drugs in their aqueous lumen are promising activatable nanocarriers for ultrasound (US)-mediated drug delivery in response to mild hyperthermia. On the other hand, US is known to locally break biological barriers and as a consequence enable internalization of molecules. In this work, a two-step protocol for intracellular delivery of cell-impermeable molecules comprising of US-induced permeabilization followed by temperature-controlled release of the model drug from thermosensitive liposomes has been developed. TSLs containing TO-PRO-3, a cell-impermeable molecule that displays a significant increase in fluorescence upon binding to nucleic acids thus serving as a 'sensor' for internalization have been prepared and characterized in detail. US-mediated permeabilization followed by temperature-controlled release was applied to tumor bearing mice following i.v. injection of TSLs and microbubbles. The efficacy of this approach was evaluated by in vivo fluorescence imaging followed by histological analysis. A 2.4-fold increase of fluorescence signal was observed and intracellular delivery of TO-PRO-3 was confirmed by a characteristic nuclear staining. These results demonstrate the feasibility of novel drug delivery system to tumors comprising of local cell permeabilization by US followed by in situ release of the payload from thermosensitive liposomes. Possible applications include local and controlled intracellular delivery of molecules with otherwise limited bioavailability.