Field study of metal concentrations and biomarker responses in resident oysters of an estuarine system in southern Brazil.
Pollutant exposure is considered an important factor responsible for the decline in marine biodiversity of Latin American coastal ecosystems. This threat has been detected in an estuarine system in southern Brazil, which prompted an investigation into the long-term biological effects of a chronic metal contamination on resident oysters from the Laguna Estuarine System (LES). Here, we present the species- and size-specific variations of biomarker responses (catalase, glucose-6-phosphate dehydrogenase, glutathione S-transferase, and protein carbonylation) in the gills and digestive gland of Crassostrea gigas and Crassostrea gasar. In parallel, concentrations of eight metals (Al, Cd, Cr, Cu, Fe, Mn, Pb, Zn) in soft tissues were measured. Our analyses revealed that the metal levels exhibited decreasing order in both species: Zn > Fe > Al > Cu > Mn > Cd. Except for Cu and Al, metal concentrations did not differ between oyster species. Biomarker results highlighted that C. gasar presented higher antioxidant responses, whereas C. gigas showed increased biotransformation upon exposure to LES pollutants, which varied according to the tissue. However, C. gasar showed a significant higher content of protein carbonylation but was not related to metals. In our research approach, the observation of metals presence and biomarkers-related responses are considered biologically relevant from an ecotoxicological perspective and serve as a baseline for future pollution studies in estuaries of Latin America. Finally, we recommend adopting a suite of biomarkers in both C. gasar and C. gigas, regardless their size and weight, as sentinel organisms in future regional biomonitoring studies in southern Brazil.


Partial caries removal in deep caries lesions: a 5-year multicenter randomized controlled trial.
This multicenter randomized controlled clinical trial aimed to compare the outcomes of stepwise excavation (SW) and partial caries removal (PCR) regarding the maintenance of pulp vitality in deep caries lesions over 5 years. At baseline, 299 permanent molars with deep caries lesions were randomly assigned to control or test groups. The control group received the stepwise excavation treatment (SW), while the test group received partial caries removal from the pulpal wall followed by restoration in a single session (PCR). Treatments were conducted in two centers located in the cities of Porto Alegre (South Brazil) and Brasília (Midwest Brazil). Survival analysis was performed to compare PCR and SW over time (Weibull regression models). The primary outcome of this study was pulp vitality, determined by the combination of the following characteristics: positive response to cold test, negative response to percussion, absence of spontaneous pain, and absence of periapical lesion (radiographic examination). This 5-year study includes data pertaining to 229 teeth: 121 teeth actually examined at the 5-year appointment, and 108 teeth contributed with data collected in previous follow-ups (18 months or 3 years). Survival analysis showed success rates of 80% in PCR group and 56% in SW group (p < 0.001). Failure was significantly associated with treatment [PCR, HR=0.38; 95%CI=0.23-0.63)] and region [South, HR=2.22; 95%CI=1.21-4.08]. PCR significantly reduced the occurrence of pulp necrosis when compared with SW. This study supports the PCR as a single-visit technique to manage deep caries lesions in permanent teeth.


Myelin and oligodendrocyte lineage cell dysfunctions: New players in the etiology and treatment of depression and stress-related disorders.
Depressive disorders are complex, multifactorial disorders that have been traditionally attributed exclusively to neuronal abnormalities. However, recent studies have increased our understanding of the contribution of glial cells-and particularly of oligodendroglia-to the pathogenesis and treatment outcome of depression and stress-related disorders. This review scrutinizes recent studies focusing on the neurosupportive functions exerted by myelin and oligodendrocyte lineage cells and their disruption in depression and stress-related disorders. It also illustrates how myelin and oligodendroglia respond to antidepressants and non-pharmacological treatment alternatives and proposes oligodendroglia-directed approaches as novel therapeutic options for depressive disorders.


Hospital outcomes after a COVID-19 diagnosis from January to May 2020 in New South Wales Australia.
To describe hospitalisation rates following COVID-19 infection in NSW. Analysis of all confirmed COVID-19 cases diagnosed in NSW from 1 January to 31 May 2020 extracted from the NSW Notifiable Conditions Information Management System and linked to routinely collected hospitalisation data. In-patient hospitalisations and hospital service utilisation details. There were 3,101 COVID-19 cases diagnosed between 1 January and 31 May 2020 in NSW: mean age 46.7 years, 50.5% were females. Overall, 12.5% (n = 389) had a record of inpatient hospitalisation, 4.2% (n = 130) were admitted to ICU and 1.9% (n = 58) received ventilation. Among adult cases, hospital and ICU admission rates increased with increasing age: 2.9% of those aged 20-29 years were hospitalised, increasing to 46.6% of those aged 80-89 years; 0.6% of those aged 20-29 years were admitted to ICU, increasing to 11.2% of those aged 70-79 years. The median time from symptoms to hospitalisation was seven days (IQR 4-11). The median time in hospital was nine days (IQR 4-20), and in ICU six days (IQR 2-15); the median time in hospital increased with older age. Almost half (49.4%) of those hospitalised with a diagnostic code had pneumonia/lower respiratory tract infection and another 36.6% had an upper respiratory tract infection or other known COVID-19 symptoms. COVID-19 is a serious infection particularly in older adults. During January to May of 2020, 1 in 8 of those diagnosed in NSW were hospitalised. While this partly reflects the cautious approach to case management in the initial phase of the pandemic, it also demonstrates the large potential impact of COVID-19 on Australian health services and need for continuing mitigation strategies.


Prevalence of Non-alcoholic Fatty Liver Disease and Steatohepatitis Risk Factors in Patients Undergoing Bariatric Surgery.
Non-alcoholic fatty liver disease (NAFLD) associated with obesity comprises pathological changes ranging from steatosis to steatohepatitis; these can evolve to cirrhosis and hepatocellular carcinoma. The objectives of this study are to assess the prevalence of and predictive markers for steatohepatitis in obese patients undergoing bariatric surgery. A prospective study of 184 morbidly obese patients undergoing bariatric surgery formed the study cohort. Patients taking potentially hepatotoxic medications and those with viral diseases and a history of excessive alcohol consumption were excluded. Liver biopsies were performed during surgery with a "Trucut" needle. Patients were classified into the following groups according to the histopathological findings: normal, steatosis, mild steatohepatitis, and moderate-severe steatohepatitis. Factors associated with steatohepatitis were evaluated using logistic regression. p values <0.05 were considered significant. The prevalence of NAFLD was 84 % (steatosis, 22.0 %; mild steatohepatitis, 30.8 %; moderate-severe steatohepatitis, 32.0 %). Independent predictive factors for steatohepatitis were age (odds ratio (OR), 1.05; 95 % confidence interval (CI), 1.01-1.09; p = 0.011), waist circumference (OR, 1.03; 95 % CI, 1.00-1.06; p = 0.021), serum alanine aminotransferase (ALT) levels (OR, 1.04; 95 % CI, 1.01-1.08; p = 0.005), and serum triglyceride levels (OR, 1.01; 95 % CI, 1.00-1.01; p = 0.042). Score values for each predictor were derived from regression coefficients and odds ratio, and a total (risk) score was obtained from the sum of the points to evaluate the probability of having steatohepatitis. Age, waist circumference, serum ALT levels, and serum triglyceride levels are efficient and non-invasive predictive markers for the diagnosis and management of steatohepatitis in morbidly obese patients.


Triterpenoid glycosides from Cephalaria transsylvanica.
On the basis of spectroscopic and chemical methods, the structures of two new triterpenoid glycosides, transsylvanoside E and F, isolated from Cephalaria transsylvanica have been established as 3-O-[beta-D-xylopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glucopyranosyl (1-->4)-beta-D-glucopyranosyl (1-->2)-beta-D-xylopyranosyl]-3 beta,23- dihydroxy delta 12-oleanen-28-carboxylic acid and 3-O-[beta-D-glucopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-beta-D-xylopyranosyl]-28-O-[beta- D-glucopyranosyl (1-->4)-beta-D-glucopyranosyl]-3 beta,23-dihydroxy delta 12-oleanen- 28-carboxylic acid, respectively. A new proglycoside was isolated from the cleavage of the ester-glycoside linkage and it's structure characterized as 3-O-[beta-D-glucopyranosyl (1-->3)-alpha-L- rhamnopyranosyl (1-->4)-beta-D-xylopyranosyl]-3 beta,23-dihydroxy delta 12-oleanen-28-carboxylic acid.


Machine learning and semi-targeted lipidomics identify distinct serum lipid signatures in hospitalized COVID-19-positive and COVID-19-negative patients.
Lipids are involved in the interaction between viral infection and the host metabolic and immunological responses. Several studies comparing the lipidome of COVID-19-positive hospitalized patients vs. healthy subjects have already been reported. It is largely unknown, however, whether these differences are specific to this disease. The present study compared the lipidomic signature of hospitalized COVID-19-positive patients with that of healthy subjects, as well as with COVID-19-negative patients hospitalized for other infectious/inflammatory diseases. We analyzed the lipidomic signature of 126 COVID-19-positive patients, 45 COVID-19-negative patients hospitalized with other infectious/inflammatory diseases and 50 healthy volunteers. A semi-targeted lipidomics analysis was performed using liquid chromatography coupled to mass spectrometry. Two-hundred and eighty-three lipid species were identified and quantified. Results were interpreted by machine learning tools. We identified acylcarnitines, lysophosphatidylethanolamines, arachidonic acid and oxylipins as the most altered species in COVID-19-positive patients compared to healthy volunteers. However, we found similar alterations in COVID-19-negative patients who had other causes of inflammation. Conversely, lysophosphatidylcholine 22:6-sn2, phosphatidylcholine 36:1 and secondary bile acids were the parameters that had the greatest capacity to discriminate between COVID-19-positive and COVID-19-negative patients. This study shows that COVID-19 infection shares many lipid alterations with other infectious/inflammatory diseases, and which differentiate them from the healthy population. The most notable alterations were observed in oxylipins, while alterations in bile acids and glycerophospholipis best distinguished between COVID-19-positive and COVID-19-negative patients. Our results highlight the value of integrating lipidomics with machine learning algorithms to explore the pathophysiology of COVID-19 and, consequently, improve clinical decision making.


Efficacy and Safety of CKD-11101 (Proposed Biosimilar of Darbepoetin-Alfa) Compared with Darbepoetin-Alfa in Patients on Hemodialysis: A Randomized, Double-Blinded, Parallel-Group Phase III Study.
Darbepoetin-alfa is an erythropoiesis-stimulating agent (ESA) with a long elimination half-life that achieves better hemoglobin (Hb) stability than short-acting ESAs. We aimed to evaluate the efficacy and safety of intravenous CKD-11101 (a biosimilar of darbepoetin-alfa) compared with those of darbepoetin-alfa in hemodialysis patients. The study was performed in 24 centers in Korea between June 2015 and June 2017. The study subjects were randomized in a double-blind manner. The follow-up duration was 24 weeks, which consisted of 20 weeks of maintenance and 4 weeks of evaluation period. All patients underwent a stabilization period to achieve a target baseline Hb of 10-12 g/dL before randomization. Following randomization, patients received darbepoetin-alfa or CKD-11101 weekly or biweekly. A total of 403 patients were randomized into two groups, and a total of 325 patients (80.6%) completed the investigation. The differences between the two groups in terms of change in the average Hb level from baseline to evaluation were not significant. The average administered dose of ESA was similar between the groups. There was no difference in the proportion of patients who maintained the target Hb during the evaluation period [60.4% vs. 66.2% in the CKD-11101 and darbepoetin-alfa groups, respectively (p = 0.3038)]. In addition, the safety analysis, consisting of adverse events and adverse drug reactions, showed comparable results between the two groups. The changes in the level of Hb, dose of erythropoietin, and achievement rate of the target Hb during the study period were comparable between the groups. CKD-11101 has an equivalent efficacy and safety compared with darbepoetin-alfa in patients undergoing hemodialysis.


Periodontal disease, hypertension, and blood pressure among older adults in Puerto Rico.
Current scientific evidence addressing the relationship between periodontitis and hypertension is limited to studies producing inconsistent results. All participants of an ongoing representative cohort of Puerto Rican elderly who were ≥70 years old and residing in the San Juan metropolitan area were invited to this cross-sectional study. Periodontal probing depth (PD) and attachment loss (AL) were summarized using the Centers for Disease Control and Prevention and the American Academy of Periodontology definition for severe periodontitis (≥2 teeth with AL ≥6 mm and ≥1 tooth with PD ≥5 mm). Three repeated blood pressure (BP) measurements taken were averaged using a standardized auscultatory method. Information on hypertension history, use of antihypertensive medications, and potential confounders (age, sex, smoking, heavy and binge drinking, diabetes, use of preventive dental services, flossing, body mass index, consumption of fruits, vegetables, whole wheat bread, and high-fiber cereal) was collected during in-person interviews. High BP was defined as average systolic BP ≥140 mm Hg or diastolic ≥90 mm Hg. Multivariate logistic regression models were used to study the relationship between severe periodontitis, hypertension history, and high BP. The study population comprised 182 adults. In multivariate analysis, there was no association between severe periodontitis and hypertension history (odds ratio [OR] = 0.99; 95% confidence interval [CI]: 0.40 to 2.48). Severe periodontitis was associated with high BP, with OR of 2.93 (95% CI: 1.25 to 6.84), after adjusting for age, sex, smoking, and binge drinking. This association was stronger when restricted to those with hypertension or taking antihypertensive medications: OR = 4.20 (95% CI: 1.28 to 13.80). The results of this study suggest that periodontitis may contribute to poor BP control among older adults.


Arthroscopic Posterior Bankart Repair Using the Wilmington Portal to Facilitate Suture Anchor Implantation.
Posterior shoulder instability (PSI) accounts for 2% to 10% of all shoulder instability, with recurrent posterior subluxation being the most common type. One of the most important risk factors is the presence of an anterior humeral notch (so-called reverse Hill-Sachs lesion), and the most common lesion in PSI is a posterior labral lesion. When conservative treatment fails, surgery is recommended to provide long-term stability, manage pain, and enable a return to previous activity levels. Most posterior labral tears are treated by an arthroscopic posterior Bankart procedure. Visualization of the posterior aspect of the glenohumeral joint is technically challenging in this procedure. The instrumental portal is also a matter of concern because there is no rotator interval posteriorly for cannula placement. The purpose of this article is to propose a technique using a secondary posterolateral Wilmington instrumental portal to perform easy and reproducible placement of the posterior suture anchor at a 45° angle to the glenoid rim. We recommend implementing this technique in patients with painful PSI or with a type B2 lesion according to the Moroder classification.


Investigation of the possibility of copper recovery from the flotation tailings by acid leaching.
The flotation tailings pond of the Bor Copper Mine poses a great ecological problem not only for the town of Bor but also for the surrounding soils and watercourses. Since the old flotation tailings contain about 0.2% of copper on the average, we investigated their leaching with sulphuric acid in the absence and presence of an oxidant. The aim was to determine the leaching kinetics of copper and iron as affected by various factors such as: the pH value of the leach solution, stirring speed, pulp density, particle size, concentration of ferric ions, temperature and time for leaching. The average copper and iron recovery obtained was from 60% to 70% and from 2% to 3%, respectively. These results indicate that the old flotation tailings pond represents an important source of secondary raw material for the extraction of copper and that it should be valorized rather than land reclamation. At the end of the paper, a mechanism of dissolution of copper and iron minerals from the tailings was described.


Antioxidant supplements: Effects on disease and aging in the United States population.
Ingestion of antioxidant supplements by the United States (US) population has increased steadily since the mid-1950's. This review tried to determine if the supplements have contributed significantly to beneficial changes in the US during this period. Experimental animal studies have demonstrated that anti-oxidant supplements lower the incidence of a wide variety of diseases and increase life span. Anti-oxidants are associated with similar changes in man. Changes since the mid-1950's in the US population include: 1) ingestion of antioxidant supplements has increased from one percent, or less, to 40-50 percent today. Cost: now 4-5 billion dollars per year, 2) disproportionate increases in the percentage of older individuals as average life expectancy at birth (ALE-B) rose, 3) declining chronic disability in the elderly since 1982, 4) declining cancer mortality since 1991, and 5) the decline in the rate of reported cardiovascular disease beginning in the 1950's which significantly increased further in 1965. The last four changes suggest that the rate of increase in physiological age with time has been slowed. This can be attributed to decreases in the rate of accumulation of free radical-induced aging changes by the joint action of antioxidant supplements/dietary measures, and improvements in conventional measures (CM) which increase ALE-B, e.g., better medical care, nutrition, housing, accident prevention. The contribution by antioxidants to decreases in physiological age is seemingly small compared to that of CM. However, it will grow relative to CM as the amount and duration of supplement use increases and improvements in CM raise ALE-B closer to 85 years, the age associated with optimal living conditions.


Follistatin-like 1 attenuates differentiation and survival of erythroid cells through Smad2/3 signaling.
Hematopoiesis is a complex process tightly controlled by sets of transcription factors in a context-dependent and stage-specific manner. Smad2/3 transcription factor plays a central role in differentiation and survival of erythroid cells. Here we report that follistatin-like 1 (FSTL1) treatment impairs hemin-induced erythroid differentiation and cell survival. FSTL1 differentially regulates transforming growth factor beta (TGF-β) and bone morphogenetic protein (BMP) signaling. Blockade of Smad2/3 signaling with the ALK5/type I TGF-βR kinase inhibitor, SB-525334, was efficacious for rescue of erythroid differentiation blockage and apoptosis. Reversely, activation of Smad1/5/8 signaling with BMP4 cannot rescue FSTL1-mediated erythroid differentiation blockage and apoptosis. Collectively, these data provide mechanistic insight into the regulation of erythropoiesis by FSTL1 signaling and lay a foundation for exploring FSTL1 signaling as a therapeutic target for anemia.


When "No-Smoking" is not enough: Hypoxia and nicotine acetylcholine receptor signaling may drive lung adenocarcinoma progression in never-smokers.
The question of how lung cancer progresses in never-smokers remains largely unanswered. In our analysis of data from 1727 lung cancer patients, we observed a difference of only 47 days in the overall survival between lung adenocarcinoma patients who were smokers vis-a-vis never-smokers - the disease has a poor prognosis irrespective of the smoking status, or gender. We have investigated the possible collaboration between the nAChR and hypoxia signaling pathway to explicate a mechanism of disease progression in never-smokers using patient-derived tumor cells. We found a previously unidentified increase in both acetylcholine and nAChR-α7 levels in non-small cell lung cancer cells in hypoxia. A similar increase in ubiquitously expressed nAChR-α7 transcripts was also observed in other cancer lines and primary tumor tissues. A direct binding of HIF-1α with the hypoxia-response element (HRE) present at -48 position preceding the transcriptional start site in nAChR-α7 promoter region was established. Crucially, the increased acetylcholine levels in hypoxia drove a feedback loop via modulation of PI3K/AKT pathway to stabilize HIF-1α in hypoxia. Further, hypoxia-mediated metastasis and induction of HIF-1α in these cells was significantly reversed by bungarotoxin, an antagonist of nAChR-α7. The nAChR-AKT-HIF network needs to be further investigated to conclusively prove its mechanism and to explore its therapeutic potential. Our study gives a plausible explanation for the equally worse prognosis of lung adenocarcinoma in never-smokers wherein the nAChR signaling is enhanced in hypoxia by acetylcholine in the absence of nicotine.


Phase I/II trial of definitive carbon ion radiotherapy for prostate cancer: evaluation of shortening of treatment period to 3 weeks.
The purpose of this study was to evaluate the feasibility of a new shortened 3-week treatment schedule of carbon ion radiotherapy (CIRT) for prostate cancer. Beginning in May 2010, patients with T1b-T3bN0M0, histologically proven prostate adenocarcinoma were enrolled in the phase II trial of CIRT. Patients received 51.6 GyE in 12 fractions over 3 weeks (protocol 1002). The primary end point was defined as the incidence of late adverse events that were evaluated based on the Common Terminology Criteria for Adverse Events version 4.0. Biochemical failure was determined using the Phoenix definition (nadir +2.0 ng ml(-1)). Forty-six patients were enrolled, and all patients were included in the analysis. The number of low-, intermediate-, and high-risk patients was 12 (26%), 9 (20%), and 25 (54%), respectively. The median follow-up period of surviving patients was 32.3 months. Two patients had intercurrent death without recurrence, and the remaining 44 patients were alive at the time of this analysis. In the analysis of late toxicities, grade 1 (G1) rectal haemorrhage was observed in 3 (7%) patients. The incidence of G1 haematuria was observed in 6 (13%) patients, and G1 urinary frequency was observed in 17 (37%) patients. No ⩾G2 late toxicities were observed. In the analysis of acute toxicities, 2 (4%) patients showed G2 urinary frequency, and no other G2 acute toxicities were observed. The new shortened CIRT schedule over 3 weeks was considered as feasible. The analysis of long-term outcome is warranted.


Catheter ablation of premature ventricular contractions originating from kissing papillary muscles.
Very thick left ventricular papillary muscles (PAMs) may kiss each other and premature ventricular contractions (PVCs) can originate from the sides of the PAMs facing each other. In such a setting, mapping of those PVCs is confusing and rendering catheter ablation challenging.


Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change.
Actin-related protein 2/3 (Arp2/3) complex is a seven-subunit assembly that nucleates branched actin filaments. Small molecule inhibitors CK-666 and CK-869 bind to Arp2/3 complex and inhibit nucleation, but their modes of action are unknown. Here, we use biochemical and structural methods to determine the mechanism of each inhibitor. Our data indicate that CK-666 stabilizes the inactive state of the complex, blocking movement of the Arp2 and Arp3 subunits into the activated filament-like (short pitch) conformation, while CK-869 binds to a serendipitous pocket on Arp3 and allosterically destabilizes the short pitch Arp3-Arp2 interface. These results provide key insights into the relationship between conformation and activity in Arp2/3 complex and will be critical for interpreting the influence of the inhibitors on actin filament networks in vivo.


A review of the treatment of primary headaches. Part II: Tension-type headache.
This paper reviews pharmacological and other approaches currently used to treat tension-type headache (TTH), and examines aspects of the classification and pathogenesis of this common complaint. Accurate diagnosis is essential before treatment is prescribed and should involve complete history taking, thorough neurological examination and evaluation of possible associated factors. The most frequently used drugs for the acute treatment of TTH are non-steroidal anti-inflammatory drugs (NSAIDs) of which only some have been shown to be efficacious in placebo-controlled trials. Amitriptyline remains the first choice treatment for prophylaxis. Other antidepressants, muscle relaxants and benzodiazepines may be used, but few have been evaluated adequately in placebo-controlled trials. Biofeedback and relaxation training, demonstrated efficacious by controlled studies, may be used when the aim is to avoid the side effects of pharmacological treatment.


Serotonin transporter (SERT) mRNA and binding site densities in male rat brain affected by sex steroids.
Estrogen increases serotonin transporter (SERT) mRNA and binding sites in female rat brain. In order to determine whether changes in SERT are gender- and steroid-specific we have now carried out studies on adult male Wistar rats which were either intact or castrated (under halothane anesthesia) and injected with arachis oil, estradiol benzoate (EB), testosterone propionate (TP) or the non-aromatizable androgen, 5alpha-dihydrotestosterone (5alpha-DHT). The number of SERT mRNA-expressing cells in the dorsal raphe (DR) nucleus was decreased by castration and increased by treatment (for approximately 32 h) with EB or TP, but not 5alpha-DHT. Sex steroids had no effect on the number of SERT mRNA-expressing cells in the median raphe nucleus. The density of SERT sites, assessed by autoradiography of [3H]paroxetine binding, was significantly reduced in arcuate nucleus and median raphe after castration, and increased in arcuate, basolateral amygdala and ventromedial hypothalamic nucleus by treatment with EB or TP, but not 5alpha-DHT. Estradiol, but not testosterone or 5alpha-DHT reduced the density of SERT sites in midbrain central grey. These data show that testosterone as well as estrogen affects SERT expression in male brain, and that the action of testosterone probably depends upon its enzymatic conversion, by aromatase, to estradiol. Our findings may have implications for sex steroid control of mood and behavior, and the action of neurotoxic derivatives of amphetamine, such as 3, 4-methylenedioxymethamphetamine, in the human.


Cutaneous leishmaniasis and leprosy.
Eight patients who had concomitant leprosy and leishmaniasis are described. Two patients with lepromatous leprosy had high resistance leishmaniasis, implying that the immune deficiency in lepromatous leprosy is specific to Mycobacterium leprae.


Topological organization of Sindbis virus capsid protein in isolated nucleocapsids.
Sindbis virus nucleocapsids were isolated from mature virions by a two-step purification method. Detergent-treated virions were sedimented in sucrose gradients and the nucleocapsid peaks chromatographed on RNase-free Sephadex G-200. The purified nucleocapsids displayed several morphologies when examined in the electron microscope. These morphologies, and the results of double-angle shadowing, suggest that the core of this enveloped virus has the shape of a regular icosahedron with a triangulation number of 4. Peptide mapping of capsid protein obtained from nucleocapsids that had been radioiodinated by a variety of means, indicated that of the four tyrosine residues in the protein, only Tyr180 was exposed at the surface of the icosahedral structure. The other three residues were not exposed on the outer surface of the nucleocapsid shell, nor on the surface of capsid protein itself, implying that they were buried within the folded protein.


Quality assessment of vitamin D metabolite assays used by clinical and research laboratories.
The Vitamin D External Quality Assessment Scheme (DEQAS) was launched in 1989 and monitors the performance of 25-hydroxyvitamin D (25-OHD) and 1,25- dihydroxyvitamin D (1,25(OH)2D) assays. In April 2015 a pilot scheme for 24,25-dihydroxyvitamin D (24,25(OH)2D) was introduced. The 25-OHD scheme is accuracy - based with target values assigned by the NIST Reference Measurement Procedure (RMP) for 25-OHD2 and 25-OHD3. A similar method is used to assign values for 3-epi-25-OHD. Five samples of human serum are distributed quarterly to over 1000 participants in 58 countries (April 2016) and clinical laboratories are expected to submit results within approximately 5 weeks. Research laboratories with assays run less frequently are not given a deadline. Archived samples with NIST- assigned values are also available. Performance is assessed on the first four samples with the fifth reserved for investigations e.g. recovery experiments or to assess the influence of other serum constituents such as lipids. DEQAS provides rapid feedback, with an on-line preliminary report available immediately after a participant submits results and a comprehensive report soon after the results deadline. In 2015, DEQAS investigations revealed that several 25-OHD immunoassays under-recovered 25-OHD2 and 25-OHD results were falsely low on a sample with a modestly raised triglyceride concentration. An RMP for 1,25 (OH)2D is not yet available and results are judged against the Method Mean. Free advice is available from the DEQAS Advisory Panel which includes experts on methodology and biostatistics. DEQAS collaborates closely with the Vitamin D Standardization Program (VDSP) and both organizations have successfully worked with participants and manufacturers to improve the accuracy of vitamin D assays.


History of cesarean before 32 weeks' gestation and trial of labor: what is the risk of uterine rupture?
To estimate the risk of uterine rupture in women with a previous cesarean delivery before 32 weeks' gestation and a trial of labor. Retrospective case control study. Setting. Level III maternity center. Among women with trial of labor, the case and control groups were women with previous cesarean section before and after 32 weeks, respectively. The main outcome was uterine rupture, defined as rupture of the three layers of the uterine wall. The study compared 108 trials of labor in patients with a previous cesarean delivery at a mean gestation of 29.8 weeks+/-2.0 SD and 137 trials of labor in patients with a previous cesarean delivery at a mean gestation of 39.2 weeks+/-2.3 SD. Mode of delivery did not differ between groups. Two uterine ruptures (1.9%, 95% CI: 0.0-4.4) occurred in the case group and none in the control group (p=0.16). Uterine ruptures caused no serious maternal or neonatal complications. The condition of the neonates was similar in the two groups. In view of the small number of uterine ruptures and the absence of severe maternal and fetal consequences over an 8-year period, a trial of labor may be proposed to women with a history of cesarean delivery before 32 weeks, when obstetrical conditions are optimal.


Differences in the ultrastructure of the submandibular glands of baboon and Rhesus monkey revealed by the use of different fixatives.
Submandibular glands of adult baboon and Rhesus monkeys were compared after different methods of fixation. In both species, serous acinar cells outnumber mucous acinar cells. In the baboon, serous cells contain secretory granules showing dense cores, moderately dense crescents, and flocculent material. In mucous cells, secretory granules vary in appearance from amorphous to highly ordered depending on fixation. In the Rhesus monkey, serous cells contain the same 3 components of secretory granules as in the baboon. Additionally, a fourth component is represented by a layer of electron-dense material between the crescent and flocculent material. Mucous cells contain electron-dense granules when fixed in Millonig's buffered fixatives, but when Clarke's or Sorensens' buffer is used the granules resemble more typical mucous granules. Duct systems of the two species are similar, but differ mainly in that large foci of glycogen are present in the striated duct cells of the Rhesus monkey.


An evaluation of the quality of commercially produced patient information leaflets.
A descriptive study was undertaken to assess the quality of a range of patient information leaflets produced by the British Dental Association. Twenty-nine leaflets were assessed with regard to presentation, readability and quality. The topic areas covered included: treatment, self-care and disease related information. Presentation was evaluated with regard to layout, font size, typeface, use of illustrations, paper type and print colour. Readability was assessed using the Flesch reading grade and the SMOG reading grade. Quality was assessed using the DISCERN tool. All leaflets scored quite well for readability, with the average SMOG Reading Grade Level being 9.10 (SD 0.80) and the average Flesch reading Grade Level being 6.18 (SD 0.83). There were, however, some areas of presentation that could be improved, specifically font size, illustration use and paper finish, which did not comply with the RNIB guidelines. Quality ratings using the DISCERN tool were low. In particular most leaflets scored poorly in setting out clear aims in the opening paragraph, in identifying sources and dates of information provided, and other sources of advice and support available. Few leaflets discussed the option of no treatment or how the treatment would affect overall quality of life. The role of shared decision making was rarely mentioned. Patient information leaflets produced commercially are of high production quality and good readability but tend not to be patient centred.


Interpretation of significance levels by psychological researchers: the .05 cliff effect may be overstated.
Comments about previous studies indicate that the interpretation of significance levels by psychological researchers is unequivocally dictated by a binary decision-making framework. In particular, confidence in a p level would drop abruptly just beyond the fateful .05 level (cliff effect). A replication of Rosenthal and Gaito's (1963) experiment on the degree of confidence in p levels shows that these claims should be moderated. Detailed analysis of individual curves reveals that the attitude of researchers toward p values is far from being as homogeneous as might be expected. However, most psychological researchers in our study rated graduated confidence judgments, as either exponential or linear. Only a minority of all-or-none respondents exhibited an abrupt drop in confidence.


Ultrastructural features of bovine mammary epithelial cells grown on collagen gels.
Bovine mammary epithelial cells cultured on floating gels of rat tail collagen showed two principal cell types, columnar and squamous, with ultrastructural features resembling secretory and myoepithelial cells respectively. Cultures of freshly prepared cells released alpha-lactalbumin into the culture medium and in some cases contained fat droplets, although these did not appear to be released. No ultrastructural evidence of casein synthesis was observed. A notable feature was the failure to secrete a continuous basement membrane. Intermediate filaments were present in abundance in squamous epithelial cells.


Cost-effectiveness analysis of molecular testing in minimally invasive samples to detect endometrial cancer in women with postmenopausal bleeding.
New approaches are being developed to early detect endometrial cancer using molecular biomarkers. These approaches offer high sensitivities and specificities, representing a promising horizon to develop early detection strategies. To evaluate the effectiveness and cost-effectiveness of introducing molecular testing to detect endometrial cancer in women with postmenopausal bleeding compared to the current strategy using the national healthcare service perspective. A Markov model was developed to assess the two early detection strategies. The model predicts the number of hysterectomies, lifetime expectancy, quality-adjusted life-years, endometrial cancer prevalence and incidence, mortality from endometrial cancer and the lifetime cost of screening, diagnosis, and treatment. Strategies were compared using the incremental cost-effectiveness ratio. The molecular strategy reduces 1.9% of the overall number of hysterectomies and the number of undetected cancer cases by 65%. Assuming a molecular test cost of 310€, the molecular strategy has an incremental cost of -32,952€ per QALY gained, being more effective and less expensive than the current strategy. The introduction of molecular testing to diagnose endometrial cancer in women presenting postmenopausal bleeding provides more health benefit at a lower cost, and therefore has the potential to be cost-effective.


Interactions of ethanol and caffeine on apoptosis in the rat cerebellum (voluntary ethanol consumers).
Ethanol alters motricity, learning, cognition, and cellular metabolism in the cerebellum. The combination of ethanol with caffeine by consuming "energy drinks" is becoming increasingly popular among young people. We analyzed the use of ethanol and caffeine on apoptosis in the cerebellum of UChB rats. The adult rats were divided into three groups (n = 14/group): UChB group: rats fed with 1:10 (v/v) ethanol ad libitum (free choice for water or ethanol) drinking from >1.9 mL of ethanol/kg body weight/day, Control group and UChB/caffeine group (free choice for water or ethanol + caffeine 300 mg/L). The treatments occurred from Day 100 till Day 150, totalizing 50 days of ethanol/caffeine ingestion. Cerebellar sections were subjected to immunohistochemistry and gene expression for Real Time-PCR (RT-PCR) for Caspase-3, XIAP, and insulin-like growth factor 1-receptor (IGF-1R). The results showed a significant increase in the gene expression of Caspase-3 and XIAP in UChB group. On the other hand, the animals of the UChB/caffeine group showed similar results to the Controls. Regarding IGFR-1, there was greater expression in the UChB groups with strong labeling in Purkinje cells. Ethanol produces neuronal and glial neurodegeneration on the cerebellum of UChB rats. The simultaneous ingestion of ethanol and caffeine reversed the ethanol damages acting caffeine with a neuroprotective effect.


Validating MEG source imaging of resting state oscillatory patterns with an intracranial EEG atlas.
Magnetoencephalography (MEG) is a widely used non-invasive tool to estimate brain activity with high temporal resolution. However, due to the ill-posed nature of the MEG source imaging (MSI) problem, the ability of MSI to identify accurately underlying brain sources along the cortical surface is still uncertain and requires validation. We validated the ability of MSI to estimate the background resting state activity of 45 healthy participants by comparing it to the intracranial EEG (iEEG) atlas (https://mni-open-ieegatlas. mcgill.ca/). First, we applied wavelet-based Maximum Entropy on the Mean (wMEM) as an MSI technique. Next, we converted MEG source maps into intracranial space by applying a forward model to the MEG-reconstructed source maps, and estimated virtual iEEG (ViEEG) potentials on each iEEG channel location; we finally quantitatively compared those with actual iEEG signals from the atlas for 38 regions of interest in the canonical frequency bands. The MEG spectra were more accurately estimated in the lateral regions compared to the medial regions. The regions with higher amplitude in the ViEEG than in the iEEG were more accurately recovered. In the deep regions, MEG-estimated amplitudes were largely underestimated and the spectra were poorly recovered. Overall, our wMEM results were similar to those obtained with minimum norm or beamformer source localization. Moreover, the MEG largely overestimated oscillatory peaks in the alpha band, especially in the anterior and deep regions. This is possibly due to higher phase synchronization of alpha oscillations over extended regions, exceeding the spatial sensitivity of iEEG but detected by MEG. Importantly, we found that MEG-estimated spectra were more comparable to spectra from the iEEG atlas after the aperiodic components were removed. This study identifies brain regions and frequencies for which MEG source analysis is likely to be reliable, a promising step towards resolving the uncertainty in recovering intracerebral activity from non-invasive MEG studies.


Noncontrast T1 mapping for the diagnosis of cardiac amyloidosis.
This study sought to explore the potential role of noncontrast myocardial T1 mapping for detection of cardiac involvement in patients with primary amyloid light-chain (AL) amyloidosis. Cardiac involvement carries a poor prognosis in systemic AL amyloidosis. Late gadolinium enhancement (LGE) cardiac magnetic resonance (CMR) is useful for the detection of cardiac amyloid, but characteristic LGE patterns do not always occur or they appear late in the disease. Noncontrast characterization of amyloidotic myocardium with T1 mapping may improve disease detection. Furthermore, quantitative assessment of myocardial amyloid load would be of great value. Fifty-three AL amyloidosis patients (14 with no cardiac involvement, 11 with possible involvement, and 28 with definite cardiac involvement based on standard biomarker and echocardiographic criteria) underwent CMR (1.5-T) including noncontrast T1 mapping (shortened modified look-locker inversion recovery [ShMOLLI] sequence) and LGE imaging. These were compared with 36 healthy volunteers and 17 patients with aortic stenosis and a comparable degree of left ventricular hypertrophy as the cardiac amyloid patients. Myocardial T1 was significantly elevated in cardiac AL amyloidosis patients (1,140 ± 61 ms) compared to normal subjects (958 ± 20 ms, p < 0.001) and patients with aortic stenosis (979 ± 51 ms, p < 0.001). Myocardial T1 was increased in AL amyloid even when cardiac involvement was uncertain (1,048 ± 48 ms) or thought absent (1,009 ± 31 ms). A noncontrast myocardial T1 cutoff of 1,020 ms yielded 92% accuracy for identifying amyloid patients with possible or definite cardiac involvement. In the AL amyloidosis cohort, there were significant correlations between myocardial T1 time and indices of systolic and diastolic dysfunction. Noncontrast T1 mapping has high diagnostic accuracy for detecting cardiac AL amyloidosis, correlates well with markers of systolic and diastolic dysfunction, and is potentially more sensitive for detecting early disease than LGE imaging. Elevated myocardial T1 may represent a direct marker of cardiac amyloid load. Further studies are needed to assess the prognostic significance of T1 elevation.


Pregnancy and epilepsy.
In a prospective study, 30 pregnant epileptic patients were followed through their pregnancy to determine the effect of seizures on pregnancy and its outcome and the effect of pregnancy on seizures. An attempt was made to correlate the serum hormone and anticonvulsant drug levels with seizure frequency, complications of pregnancy, occurrence of status epilepticus and teratogenicity. In 14 patients seizure frequency increased, in 15 it remained unchanged and in 1 patient it decreased. There were 2 spontaneous abortions, 2 patients had status epilepticus and 1 offspring of a patient had a ventricular septal defect. This patient was receiving carbamazepine and diphenylhydantoin. Patients with increased seizures frequency had significantly higher oestrogen levels, lower level of progesterone and lower level of anticonvulsant drugs as compared with those with no change in seizures. Patients who had abortions and those who developed status epilepticus had high serum oestrogen levels.


Serum concentrations of CRP, IL-6, TNF-α and cortisol in major depressive disorder with melancholic or atypical features.
The aim of the present study was to explore possible differences between serum C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), and cortisol concentration in patients with major depressive disorder (MDD) with melancholic features, and MDD with atypical features. As secondary aim, we investigated possible associations with clinical features such as suicidal ideation, number of episodes, duration of depression and symptomatology severity. We included 55 MDD patients (32 with melancholic features and 23 with atypical features) and 18 healthy controls. When compared to healthy controls, MDD with melancholic or atypical features showed higher CRP and IL-6, but not TNF-α. Cortisol concentration was higher in MDD with melancholic type, in comparison to the atypical type of MDD or controls. A positive correlation was found between the severity of depressive symptoms, concentrations of IL-6 and cortisol in the MDD group with melancholic features, while a negative correlation was observed between IL-6 and CRP in the MDD group with atypical feature. Also, in the MDD group with atypical features, there was a correlation between the severity of anxiety symptoms based on the Hamilton Rating Scale for Anxiety (HARS), concentration of CRP, and the duration of symptoms. In conclusion, we observed several differences in serum CRP, IL-6, and cortisol concentrations in MDD patients considering clinical features as well.


[The Impact of the COVID-19 Pandemic on the Intensive Care Residency Program in Portugal].
In 2020, critical care departments underwent profound changes imposed by the COVID-19 pandemic. The aim of this study was to evaluate the impact of the pandemic on the intensive care residency program in Portugal. The Association of Critical Care Residents (AIMINT) prepared a questionnaire using the Google Forms® tool, which was applied during August 2020 to the Critical Care residents in Portugal. A descriptive analysis was performed with the data collected. Eighty-five residents participated in the questionnaire, yieldinga response rate of 62%. Three-quarters of all participants provided care to COVID-19 patients. More than 80% of the surveyed participants were on rotations, and these were canceled in 59% of cases. Seventy-eight percent reported a workload greater than 40 hours per week. The COVID-19 pandemic had an impact on the Critical Care Residency program in Portugal. Most residents surveyed provided care to COVID-19 patients and not only saw their rotations suspended but also experienced difficulties in rescheduling them.


High-throughput genotyping of a full voltage-gated sodium channel gene via genomic DNA using target capture sequencing and analytical pipeline MoNaS to discover novel insecticide resistance mutations.
In insects, the voltage-gated sodium channel (VGSC) is the primary target site of pyrethroid insecticides. Various amino acid substitutions in the VGSC protein, which are selected under insecticide pressure, are known to confer insecticide resistance. In the genome, the VGSC gene consists of more than 30 exons sparsely distributed across a large genomic region, which often exceeds 100 kbp. Due to this complex genomic structure, it is often challenging to genotype full coding nucleotide sequences (CDSs) of VGSC from individual genomic DNA (gDNA). In this study, we designed biotinylated oligonucleotide probes from CDSs of VGSC of Asian tiger mosquito, Aedes albopictus. The probe set effectively concentrated (>80,000-fold) all targeted regions of gene VGSC from pooled barcoded Illumina libraries each constructed from individual A. albopictus gDNAs. The probe set also captured all orthologous VGSC CDSs, except some tiny exons, from the gDNA of other Culicinae mosquitos, A. aegypti and Culex pipiens complex, with comparable efficiency as a result of the high nucleotide-level conservation of VGSC. To improve efficiency of the downstream bioinformatic process, we developed an automated pipeline-MoNaS (Mosquito Na+ channel mutation Search)-which calls amino acid substitutions in the VGSC from NGS reads and compares those to known resistance mutations. The proposed method and our bioinformatic tool should facilitate the discovery of novel amino acid variants conferring insecticide resistance on VGSC and population genetic studies on resistance alleles (with respect to the origin, selection, and migration etc.) in both clinically and agriculturally important insect pests.


The measurement and mass spectral identification of indole-3-pyruvate from tomato shoots.
Endogenous indole-3-pyruvate has been identified by full-scan combined gas chromatography-mass spectrometry and measured using gas chromatography with an electron capture detector. High specific-activity [5-3H]indole-3-pyruvate was synthesized from [5-3H]tryptophan and used as an internal standard. In order to allow purification of the labile indole-3-pyruvate it was stabilised by the formation in the crude extract of its pentafluorobenzyl oxime derivative. This derivative also allowed sensitive detection and measurement of indole-3-pyruvate in the picogram range using a gas chromatograph with an electron capture detector. Endogenous levels were found to be between 8-10 ng/g f.wt. of tomato shoots which is comparable to that of the indole-3-acetic acid pool size, 11 ng/g f.wt., in this tissue.


Author Correction: Bacterial community analysis of floor dust and HEPA filters in air purifiers used in office rooms in ILAS, Beijing.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Probing GPCR structure: adenosine and P2Y nucleotide receptors.
The adenosine receptors (ARs) provide an example of how to accurately predict ligand recognition, even prior to the availability of a crystallographic structure. Homology modeling has been used to gain structural insight, in conjunction with site-directed mutagenesis, and structure-activity relationships of small molecular ligands. Recent X-ray structures greatly improved the accuracy of knowledge of AR ligand recognition and furthermore characterized conformational changes induced by receptor activation. Now, homology modeling extends these structural insights to related GPCRs and suggests new ligand structures. This strategy is also being applied to the eight subtypes of P2Y receptors for extracellular nucleotides, which lack X-ray structures and are best modeled by homology to the CXCR4 (peptide) receptor. Neoceptors, as studied for three of the four AR subtypes, create a molecular complementarity between a mutant receptor and a chemically tailored agonist ligand to selectively enhance affinity, implying direct physical contact and thus validating docking hypotheses.


Cholesterol sulfate inhibits osteoclast differentiation and survival by regulating the AMPK-Sirt1-NF-κB pathway.
Cholesterol sulfate (CS) is an activator of retinoic acid-related orphan receptor α (RORα). CS treatment or RORα overexpression attenuates osteoclastogenesis in a collagen-induced arthritis mouse model. However, the mechanism by which CS and RORα regulate osteoclast differentiation remains largely unknown. Thus, we aimed to investigate the role of CS and RORα in osteoclastogenesis and their underlying mechanism. CS inhibited osteoclast differentiation, but RORα deficiency did not affect osteoclast differentiation and CS-mediated inhibition of osteoclastogenesis. CS enhanced adenosine monophosphate-activated protein kinase (AMPK) phosphorylation and sirtuin1 (Sirt1) activity, leading to nuclear factor-κB (NF-κB) inhibition by decreasing acetylation at Lys310 of p65. The NF-κB inhibition was restored by AMPK inhibitor, but the effects of CS on AMPK and NF-κB were not altered by RORα deficiency. CS also induced osteoclast apoptosis, which may be due to sustained AMPK activation and consequent NF-κB inhibition, and the effects of CS were significantly reversed by interleukin-1β treatment. Collectively, these results indicate that CS inhibits osteoclast differentiation and survival by suppressing NF-κB via the AMPK-Sirt1 axis in a RORα-independent manner. Furthermore, CS protects against bone destruction in lipopolysaccharide- and ovariectomy-mediated bone loss mouse models, suggesting that CS is a useful therapeutic candidate for treating inflammation-induced bone diseases and postmenopausal osteoporosis.


Effectiveness of martial arts exercise on anthropometric and body composition parameters of overweight and obese subjects: a systematic review and meta-analysis.
Obesity is considered a top public health concern, and its prevalence is growing every day. Thus, interventions to address this problem should be encouraged and further studied. In this regard, the aim of this review was to summarize the evidence of martial arts interventions to evaluate their effectiveness on the anthropometric and body composition parameters of overweight and obese subjects. A systematic literature search was conducted on January 26, 2020 using the PubMed, Medline, Lilacs, Cochrane, and Scielo databases. Reference lists of eligible articles and relevant reviews have also been examined. All randomized clinical trials on martial arts that evaluated the anthropometric and body composition parameters of overweight and obese subjects were included, and a narrative synthesis of eligible studies was conducted in accordance with PRISMA guidelines. The Downs & Black checklist was used to assess the quality of the studies. This review was registered in the International Prospective Register of Systematic Reviews (PROSPERO) (identifier CRD42018086116). A total of 82 articles were identified from the initial search strategy. A further 2 articles were identified from the review of relevant bibliographies. Six studies encompassing 258 participants who were overweight or obese were included. Four studies reported Tai Chi practice, one study reported Kung Fu exercise, and another study reported martial arts exercise. The examined meta-analyses did not reveal significant benefits from martial arts practice over control groups after the experiment period for body mass index (- 1.34 kg/m2; 95% CI: - 2.72, 0.05), waist circumference (1.41 cm; 95% CI: - 0.72, 3.54) and percentage of body fat (- 0.75%; 95% CI: - 5.58, 4.08). The scarcity, heterogeneity, short intervention time, small sample size, and significant methodological limitations of the available studies do not allow to conclude whether martial arts are effective in the anthropometric and body composition parameters of overweight and obese individuals. This study highlights the need for more research to assess the benefits of martial arts for overweight and obese individuals.


Natural cytotoxicity in malignant and premalignant cervical neoplasia and enhancement of cytotoxicity with interferon.
Natural cytotoxicity was measured in 70 women with cervical dysplasia or invasive cervical cancer and in 13 controls, using a short-term chromium release assay. A significant diminution in mean cytotoxicity was found in women with CIN III and with advanced cervical carcinoma. When the peripheral blood leukocytes of patients and controls were pretreated with interferon, there was significant enhancement of the mean cytotoxicity for all groups except those with advanced cervical carcinoma (stage IIB or worse). Our results have shown a significant defect in a potentially important immune surveillance mechanism in a group of patients with premalignant disease at risk for the development of invasive cancer. A possible role for interferon in the treatment of cervical intraepithelial neoplasia is suggested.


Biomechanical competence of vertebral trabecular bone in relation to ash density and age in normal individuals.
Cylindrical trabecular bone samples from the central part of the first lumbar vertebral body were obtained from 42 normal people (27 females and 15 males) aged 15-87 years and analyzed by a compression test in either vertical or horizontal direction. Maximum stress, maximum stiffness, energy absorption capacity, and strain at maximum stress (compressibility) were calculated from the load-deformation curves. The ash density of the samples was measured after incineration. From age 20 to 80 years, the decline (P less than 0.01) in ash density was 48-50% compared to a 75-80% decrease (P less than 0.001) in vertical stress, stiffness, and energy absorption capacity. Qualitatively similar age-related changes (P less than 0.01) were observed by horizontal compression, but the absolute values were smaller. In both directions, highly significant positive correlations (P less than 0.01) were observed between the biomechanical properties and ash density stress, stiffness and energy absorption capacity still showed significant decreases with age (P less than 0.01). The vertical maximum strain values, which increased (P less than 0.05) with age, were inversely related to the other biomechanical variables (P less than 0.05) and to the ash density (P less than 0.05). It is concluded from the study that the biomechanical competence of vertebral trabecular bone depends not only on bone mass (ash density) but also on the continuity of the trabecular lattice, which changes with increasing age.


An effective approach for BRAF V600E mutation analysis of routine thyroid fine needle aspirates.
Molecular testing for genetic alterations in thyroid neoplasms, including BRAF V600E (BRAF) mutation, are often applied to thyroid aspirates falling into the Bethesda System for Reporting Thyroid Cytopathology indeterminate categories. Current methods typically use dedicated aspirated material, without morphological determination of containing the cells of interest and may be of elevated cost. We describe our experience with BRAF mutation analysis on material obtained from Papanicolaou (PAP)-stained ThinPrep® (TP) slides. Eighty-three cases collected between 2012 and 2019 with more than 100 cells were selected. An electronic record of a whole slide scan was made for each case before testing. The coverslips were removed, and DNA was extracted from material scraped from each slide using the Qiagen QIAamp DNA FFPE Tissue Kit. BRAF testing was performed using a highly sensitive mutation detection assay, either COLD-PCR, castPCR, or droplet digital PCR. Fourteen out of 83 cases had a BRAF mutation. Of these, 8 were classified as atypia of undetermined significance or suspicious for malignancy in which follow-up showed conventional papillary thyroid carcinoma in 5 out of 6 cases. The specificity and positive predictive value were 97% and 91%, respectively. BRAF mutation analysis can be performed on material obtained from routine clinical PAP-stained TP slides. As a first step, this unconventional effective approach may reduce costs related to the molecular evaluation of thyroid nodule aspirates and provides the opportunity for cytomorphological confirmation that the cells of interest are present in material submitted for BRAF mutation analysis.


Kinetic pulse-labeling study of Fusarium culmorum. Biosynthetic intermediates and dead-end metabolites.
A kinetic pulse-labeling method was utilized in Fusarium culmorum to detect plausible biosynthetic intermediates and differentiate them from dead-end metabolites. The ultimate test to demonstrate a precursor relies on feeding experiments. We now report the detection of four new metabolites, one of them (compound 1) behaves as a dead-end metabolite, whereas compounds 2, 3, and 4 seem to be putative intermediates: they metabolize with time just when 3-acetyldeoxynivalenol (3-ADN) and/or sambucinol (SOL) start to be produced. Feeding experiments confirmed these results: compound 1 is not converted to 3-ADN or SOL, and compounds 2-4 are precursors to 3-ADN. In addition 3 is a precursor to SOL.


Determination of volatile organic compounds in water by headspace solid-phase microextraction gas chromatography coupled to tandem mass spectrometry with triple quadrupole analyzer.
In the present work, a rapid method with little sample handling has been developed for determination of 23 selected volatile organic compounds in environmental and wastewater samples. The method is based on headspace solid-phase microextraction (SPME) followed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS) determination using triple quadrupole analyzer (QqQ) in electron ionization mode. The best conditions for extraction were optimised with a factorial design taking into account the interaction between different parameters and not only individual effects of variables. In the optimized procedure, 4 mL of water sample were extracted using a 10 mL vial and adding 0.4 g NaCl (final NaCl content of 10%). An SPME extraction with carboxen/polydimethylsiloxane 75 μm fiber for 30 min at 50°C (with 5 min of previous equilibration time) with magnetic stirring was applied. Chromatographic determination was carried out by GC-MS/MS working in Selected Reaction Monitoring (SRM) mode. For most analytes, two MS/MS transitions were acquired, although for a few compounds it was difficult to obtain characteristic abundant fragments. In those cases, a pseudo selected reaction monitoring (pseudo-SRM) with three ions was used instead. The intensity ratio between quantitation (Q) and confirmation (q) signals was used as a confirmatory parameter. The method was validated by means of recovery experiments (n=6) spiking mineral water samples at three concentration levels (0.1, 5 and 50 μg L(-1)). Recoveries between 70% and 120% were generally obtained with relative standard deviations (RSDs) lower than 20%. The developed method was applied to surface water and wastewater from a wastewater treatment plant and from a municipal solid-waste treatment plant. Several compounds, like chloroform, benzene, trichloroethylene, toluene, tetrachloroethylene, dibromochloromethane, xylenes and bromoform were detected and confirmed in all the samples analyzed.


The role of gamified learning strategies in student's motivation in high school and higher education: A systematic review.
Gamification, defined as the integration of videogame components to promote a gameful experience, is increasingly being implemented in education with the aim of enhancing students' engagement and motivation. Accordingly, since 2010 it has constituted an area of growing interest for researchers and teachers. Following PRISMA 2020's methodology, a systematic review (SR) was conducted in November 2022 seeking to explore the influence of gamification strategies on students' motivation to learn. Having identified 548 articles, 40 studies were chosen based on the selection criteria set and analyzed to reveal that game elements such as points, badges and rankings are widely used to motivate students. From a theoretical perspective, gamification studies focus on the dichotomy of intrinsic and extrinsic motivation. The results suggest a positive influence of gamification strategies on students' motivation, although in the long run, such motivation can decline. Furthermore, the influence of a novelty effect and extrinsic rewards on motivation is identified, which can lead to greater motivation in the short term, followed by a decrease with further exposure to gamification. Future studies should focus on the influence of students' individual traits (e.g., gaming experience, openness to competition and cooperation) on gamification strategies. Moreover, long-term exposure to gamification as well as the novelty effect should be explored.


Recognizing the advantage of employing the PowerPlex(®) ESI and PowerPlex(®) ESX complementary kits for purposes of clarification in routine casework.
In an effort to promote European cross-border cooperation in fighting crime and international terrorism the Treaty of Prüm was drafted and accepted within the European forensic community. This move led to the commercial development of new multiplex kits which introduced five new STR loci and promised better performance. Recently the Israel Police DNA Casework and Database laboratories adopted the PowerPlex(®) ESI kit for routine use in our laboratories. Presented in this paper are examples of three types of ambiguous results encountered during the implementation of the PowerPlex(®) ESI kit into routine work. These ambiguous products presented themselves in the form of (1) extreme variants outside of loci borders, (2) failure to amplify sister allele pairs or expression of null alleles and (3) episodes of loss of separation of adjacent microvariants primarily in mixture samples. The re-analysis of all these samples using the PowerPlex(®) ESX kit successfully and rapidly clarified all three categories of anomalies. Spotlighting such events to the forensic community, especially regarding the novel loci introduced in these next generation kits, can aid in raising the analyst's awareness to their future appearances and prevent possible erroneous conclusions. In addition, providing timely DNA results to investigating teams is of great importance and operational forensic laboratories do not have at their immediate disposal methods such as sequencing to elucidate such manifestations. We suggest that the complementary use of the PowerPlex(®) ESI and PowerPlex(®) ESX can provide a benefit for clarification purposes in routine casework.


Comparable immunoreactivity rates of PD-L1 in archival and recent specimens from non-small cell lung cancer.
Molecular targeted therapy including the use of monoclonal antibodies directed against the immune checkpoints PD-L1 and PD-1 receptor have remarkably improved the therapeutic response and survival of cancer patients. The tumor expression level of PD-L1 can predict the response rate to checkpoint inhibitors. We evaluated whether the time interval between tumor tissue sampling/paraffinization and immunohistochemistry affects the staining level of PD-L1 in non-small cell lung cancer (NSCLC). This study comprised 137 patients with NSCLC. Tumors were stained with 22C3 or 28-8 antibodies. There was a significant correlation between the immunoreactivity rate of tumor tissues obtained using 22C3 and 28-8 clones. No statistical difference in immunoreactivity between archival and recent samples stained either with 22C3 or 28-8 antibodies was observed. The immunoreactivity rate achieved with 22C3 or 28-8 antibodies significantly correlated with tumor histological type and size, but not with specimen storage time, age, gender, smoking history, clinical stage, or lymph node metastasis. In brief, the results of this study show that the time interval between tissue sampling/paraffinization and immunohistochemical analysis has no influence on the immunoreactivity rate of PD-L1 in NSCLC.


Universal model for accurate calculation of tracer diffusion coefficients in gas, liquid and supercritical systems.
In this work it is presented a new model for accurate calculation of binary diffusivities (D12) of solutes infinitely diluted in gas, liquid and supercritical solvents. It is based on a Lennard-Jones (LJ) model, and contains two parameters: the molecular diameter of the solvent and a diffusion activation energy. The model is universal since it is applicable to polar, weakly polar, and non-polar solutes and/or solvents, over wide ranges of temperature and density. Its validation was accomplished with the largest database ever compiled, namely 487 systems with 8293 points totally, covering polar (180 systems/2335 points) and non-polar or weakly polar (307 systems/5958 points) mixtures, for which the average errors were 2.65% and 2.97%, respectively. With regard to the physical states of the systems, the average deviations achieved were 1.56% for gaseous (73 systems/1036 points), 2.90% for supercritical (173 systems/4398 points), and 2.92% for liquid (241 systems/2859 points). Furthermore, the model exhibited excellent prediction ability. Ten expressions from the literature were adopted for comparison, but provided worse results or were not applicable to polar systems. A spreadsheet for D12 calculation is provided online for users in Supplementary Data.


Tailored Sailing Experience to Reduce Psychological Distress and Improve the Quality of Life of Breast Cancer Survivors: A Survey-Based Pilot Study.
Background: Growing evidence indicates that physical/sporting activities may improve the health outcomes and quality of life (QoL) of breast cancer (BC) survivors. Since recent reports have suggested that sailing can improve the psychophysical well-being and QoL of people with disabilities, this pilot study evaluated the effectiveness of a tailored sailing experience on the QoL and psychological distress (PD) of BC survivors. Methods: A group of 19 breast cancer survivors, who were attending the Cancer Rehabilitation Center in Florence, were invited to participate in a sailing school and completed a survey based on a structured online questionnaire assessing QoL and PD both on departure (baseline) and one week after returning (follow-up). The survey comprised a first part (i.e., sociodemographic characteristics and the practice of physical/sporting activities at baseline; sailing experience satisfaction at follow-up) and a second part (i.e., Short Form-12 (SF-12), State/Trait-Anxiety Inventory form Y (STAI-Y), distress thermometer questionnaires). A paired Student's t-test was used to compare the baseline versus follow-up QoL and PD scores. Results: A statistically significant improvement in SF-12 mental component scores and a reduction in both STAI-Y state/trait components and distress thermometer scores were found after the sailing experience. Conclusions: We conclude that sailing practice could be a feasible intervention to increase the psychophysical well-being of BC survivors.


Life cycle assessment of photovoltaic electricity production by mono-crystalline solar systems: a case study in Canada.
Photovoltaic (PV) system is widely recognized as one of the cleanest technologies for electricity production, which transforms solar energy into electrical energy. However, there are considerable amounts of emissions during its life cycle. In this study, life cycle assessment (LCA) was used to evaluate the environmental and human health impacts of PV electricity production in Canada. The PV potential varies considerably among the provinces, with higher values in Manitoba (MB), Saskatchewan (SK), Alberta (AB), and southern Ontario (ON). A grid-connected slanted-roof mono-crystalline silicon (mono-Si) PV system with a capacity of 3 kWp (the peak power of the system in kilowatts) in Toronto, Ontario, was considered as the case study system. Ten impact categories were considered including (1) acidification, (2) carcinogenic, (3) ecotoxicity, (4) eutrophication, (5) fossil fuel depletion, (6) global warming, (7) non-carcinogenic, (8) ozone depletion, (9) respiratory effects, and (10) smog. Among the four components of the PV system, i.e., mono-Si panel, mounting system, inverter, and electric installation, the mono-Si panel production was the highest contributor in seven out of ten impact categories, including acidification (68%), eutrophication (60%), fossil fuel depletion (81%), global warming (77%), ozone depletion (88%), respiratory effects (74%), and smog (70%). For the other three processes, the electric installation contributed most to ecotoxicity at 58%, followed by the mounting system in the carcinogenic category (29%), and the inverter in the non-carcinogenic category (31%). By normalizing the impacts based on the reference scores in Canada, it was found that the ecotoxicity and carcinogenic categories had dominant contributions to the overall impact by 53% and 42%, respectively. The global warming potential impact was estimated as 79 gr CO2 eq /kWh, which is close to the mean value of 79.5 gr CO2 eq /kWh, reported in the literature. The sensitivity analysis indicated that a 10% increase in the panel and mounting system area will increase the ozone depletion and carcinogenic categories by 8.1% and 2.8%, respectively.


Transformation of Bacillus thuringiensis plasmid DNA by a new polyethylenimine polymeric nanoparticles method.
Although electroporation technique has been mostly used to transform Bacillus thuringiensis (Bt), this method is not readily applicable to strains other than the one for which it was optimized. Polyethylenimine (PEI) is a golden standard non-viral vector that interacts with plasmids to form compact polymeric nanoparticles (PNPs) via electrostatic interactions. This PNPs system is very attractive because they are easily prepared, able to carry large nucleic acid constructs, and show low toxicity. In this study, PEI/pBTdsSBV-VP1 PNPs were successfully prepared at various N/P ratios which is positively-chargeable polymer amine (N = nitrogen) groups to negatively-charged nucleic acid phosphate (P) groups, and the internalization of the complexes into Bt 4Q7 was confirmed by confocal laser scanning microscopy. The PEI-mediated transformation showed similar efficiency comparable to that of electroporation method, suggesting that the method of PNPs will be an effective alternative for transformation of Bt strains.


Quantifying microplastics pollution in the Red Sea and Gulfs of Suez and Aqaba: Insights from chemical analysis and pollution load assessment.
Microplastics (MPs) constitute the majority of marine plastic litter. The pollution caused by MPs has been categorized as a gradual and persistent crisis, but little is known about its extent along the shores of the Red Sea, particularly on the Egyptian side. The Red Sea is a rapidly developing region and home to critical ecosystems with high levels of endemism. This study represents the first comprehensive survey investigating the extent of MP pollution along the Egyptian shores of the Red Sea, including the Gulf of Suez and Aqaba. Mean concentrations ranged from 23.3 ± 15.28 to 930.0 ± 181.9 MPs/kg DW. Out of 17 beaches surveyed, 12 had mean concentrations of <200 items/kg, indicating a low occurrence of MPs compared to the shores of the Mediterranean Coast of Egypt. The pollution load index varied from low to medium levels in most locations. Ras Mohamed, a marine protected area, showed high vulnerability to MP pollution. All the investigated particles were fragments of secondary MPs. The sources of pollution mainly come from maritime activities, including cargo shipping and intense recreational activities. Fourier Transform Infrared Spectroscopy identified four plastic polymers, with polyethylene and polypropylene being the most common. The surface morphology of plastic particles was examined using scanning electron microscopy combined with energy-dispersive X-ray spectroscopy. All the particles exhibited signs of degradation, which could generate countless plastic pieces with possible deleterious impacts. This work has highlighted the importance of conducting region-specific assessments of mismanaged plastic waste, focusing on the role of tourism and recreational navigation as contributors to plastic litter, to estimate plastic waste inputs into the waters of the Red Sea Coast of Egypt. Efforts are needed to develop strategic plans to reduce the disposal of plastic waste in the region.


Freeze-dried synthesized bifunctional biopolymer nanocomposite for efficient fluoride removal and antibacterial activity.
Local fluoride contamination and bacterial infections in potable water have dangerous effects on the human body and are today a global concern. In this study, we have synthesized a pH-responsive bifunctional biopolymer nanocomposite (HAZ) of humic acid with incorporating aluminum zirconium bimetallic oxide by deep freeze-drying method. Fast nucleation and interconnection of nanoparticles form a highly porous network because of sublimation of frozen HAZ. This duo nanocomposite has efficiently worked for fluoride removal and showed potent antibacterial activity against the Escherichia coli Gram-negative and Staphylococcus aureus Gram-positive bacteria. The X-ray photoelectron spectroscopy (XPS) analysis demonstrates that the hydroxyl groups act as a pivot in the ion exchange process of adsorption, each element of bimetallic oxide primarily takes part in the adsorption mechanism. The maximum adsorption capacity of the adsorbent was 180.62 mg/g at pH seven. Thermodynamic parameters like Gibbs free energy change (ΔG0), entropy (ΔS0), and enthalpy (ΔH0) indicate that the process was endothermic, feasible, and taken place by a chemisorption mechanism. This is the first novel freeze-dried bifunctional biopolymer nanocomposite composed of humic acid natural polymer incorporated with Al-Zr metal oxide, and it exhibited three times higher adsorption efficacy with excellent antibacterial action at a concentration of 5 µg/mL of the nanocomposite.


Research advances in identification procedures of endocrine disrupting chemicals.
Endocrine disrupting chemicals (EDCs) are increasingly concerned substance endangering human health and environment. However, there is no unified standard for identifying chemicals as EDCs, which is also controversial internationally. In this review, the procedures for EDC identification in different organizations/countries were described. Importantly, three aspects to be considered in identifying chemical substances as EDCs were summarized, which were mechanistic data, animal experiments, and epidemiological information. The relationships between them were also discussed. To elaborate more clearly on these three aspects of evidence, scientific data on some chemicals including bisphenol A, 1,2-dibromo-4-(1,2 dibromoethyl) cyclohexane and perchlorate were collected and evaluated. Altogether, the above three chemicals were assessed for interfering with hormones and elaborated their health hazards from macroscopic to microscopic. This review is helpful for standardizing the identification procedure of EDCs.


Isolation of a replication-defective murine leukaemia virus from cultured AKR leukaemia cells.
Mice of the AKR strain are characterised by a high incidence of spontaneous thymic lymphomas. AKR chromosomes contain the genomes of ecotropic murine leukaemia virus (MuLV) at two loci, termed Akv-1 and Akv-2 (refs 2-6). Shortly after birth, the normal tissues of AKR mice begin to produce high levels of this XC-positive MuLV (ref. 7) (that is, one that forms XC plaques). A second class of MuLV, termed mink cell focus-inducing virus (MCF), is produced specifically by preleukaemic and leukaemic AKR thymocytes. Nowinski et al. have established a series of tissue culture lines from AKR leukaemias and reported that the resulting cell lines produce virus particles, but that these particles, surprisingly, do not give rise to XC plaques. We have analysed the virus particles produced by one of these cell lines, termed AKRSL2. We show here that, unlike most or all of the nonmalignant tissues in the AKR mouse, these cultured lymphoma cells produce very little non-defective ecotropic MuLV; however, they do produce replication-defective ecotropic MuLV.


Response of invertebrates to lotic disturbance: a test of the hyporheic refuge hypothesis.
Recovery following hydrological disturbances is usually rapid for lotic invertebrates. Stream ecologists have assumed that recovery is facilitated by behavioral migrations during floods down into the hyporheic zone (the interstitial spaces of a streambed) to seek temporary refuge from possible erosion (the "hyporheic refuge hypothesis"). We provide the first explicit test of this hypothesis by evaluating three predictions of the hypothesis. We coupled field observations of the response of meiofaunal invertebrates to floods with field and flume experiments. The study site was a sandy-bottom stream in northern Virginia. Prediction 1, that loss of fauna from a streambed during floods should be minimal as long as the depth of scour in the streambed is less than the depth of the hyporheic zone, was not supported for any taxon. For two floods which varied considerably in magnitude, 50-90% of the fauna was lost from the bed despite the fact that the depth of scour (10-30 cm) was significantly less than the total depth of the hyporheic zone (50 cm). Prediction 2, that fauna should move deeper into the bed at higher flows, was supported by field observations during only one of two floods and then only for rotifers. In flume experiments that tested for finer scale behavioral movements, significant vertical migrations were found for copepods and chironomids which moved 1.5-3.5 cm downward as mean velocity (3 cm off bottom) was increased from 5-23 cm/s. Movements down by rotifers were not found in the flume experiments. Prediction 3, that the hyporheic zone is the most important source of colonists to defaunated areas, was supported in part by field experiments. The hyporheic route was not the primary route for any taxon but it was as important for the rotifers and copepods as water column or streambed surface routes. We conclude that, even though smallscale (cm's) migrations into the streambed in response to increased flow may be observed for some taxa and the hyporheic zone may serve as a partial source of colonists following disturbances, movements down are not adequate in preventing significant losses of meiofauna during floods.


Umbilical Cord Tissue Offers the Greatest Number of Harvestable Mesenchymal Stem Cells for Research and Clinical Application: A Literature Review of Different Harvest Sites.
Recent years have seen dramatic increases in the techniques used to harvest and isolate human mesenchymal stem cells. As the potential therapeutic aspects of these cells further develop, informative data on the differences in yields between tissue harvest sites and methods will become increasingly valuable. We collected and compared data on cell yields from multiple tissue harvest sites to provide insight into the varying levels of mesenchymal stem cells by tissue and offer primary and alternative tissue types for harvest and clinical application. The PubMed and Medline databases were searched for articles relating to the harvest, isolation, and quantification of human mesenchymal stem cells. Selected articles were analyzed for relevant data, which were categorized according to tissue site and, if possible, standardized to facilitate comparison between sites. Human mesenchymal stem cell levels in tissue varied widely according to tissue site and harvest method. Yields for adipose tissue ranged from 4,737 cells/mL of tissue to 1,550,000 cells/mL of tissue. Yields for bone marrow ranged from 1 to 30 cells/mL to 317,400 cells/mL. Yields for umbilical cord tissue ranged from 10,000 cells/mL to 4,700,000 cells/cm of umbilical cord. Secondary tissue harvest sites such as placental tissue and synovium yielded results ranging from 1,000 cells/mL to 30,000 cells/mL. Variations in allogeneic mesenchymal stem cell harvest levels from human tissues reflect the evolving nature of the field, patient demographic characteristics, and differences in harvest and isolation techniques. At present, Wharton's jelly tissue yields the highest concentration of allogeneic mesenchymal stem cells whereas adipose tissue yields the highest levels of autologous mesenchymal stem cells per milliliter of tissue. This comparison of stem cell levels from the literature offers a primer and guide for harvesting mesenchymal stem cells. Larger mesenchymal stem cell yields are more desirable for research and clinical application.


A nanocarrier system that potentiates the effect of miconazole within different interkingdom biofilms.
Novel and new therapeutic strategies capable of enhancing the efficacy of existing antimicrobials is an attractive proposition to meet the needs of society. This study aimed to evaluate the potentiating effect of a miconazole (MCZ) nanocarrier system, incorporated with iron oxide nanoparticles (IONPs) and chitosan (CS) (IONPs-CS-MCZ). This was tested on three representative complex interkingdom oral biofilm models (caries, denture and gingivitis). The planktonic and sessile minimum inhibitory concentrations (MICs) of IONPs-CS-MCZ against different Candida albicans strains were determined, as well as against all represented bacterial species that formed within the three biofilm models. Biofilms were treated for 24 hours with the IONPs-CS nanocarrier system containing MCZ at 64 mg/L, and characterized using a range of bioassays for quantitative and qualitative assessment. MIC results generally showed that IONPs-CS-MCZ was more effective than MCZ alone. IONPs-CS-MCZ also promoted reductions in the number of CFUs, biomass and metabolic activity of the representative biofilms, as well as altering biofilm ultrastructure when compared to untreated biofilms. IONPs-CS-MCZ affected the composition and reduced the CFEs for most of the microorganisms present in the three evaluated biofilms. In particular, the proportion of streptococci in the biofilm composition were reduced in all three models, whilst Fusobacterium spp. percentage reduced in the gingivitis and caries models, respectively. In conclusion, the IONPs-CS-MCZ nanocarrier was efficient against three in vitro models of pathogenic oral biofilms, showing potential to possibly interfere in the synergistic interactions among fungal and bacterial cells within polymicrobial consortia.


Resolution of fear of flying with fluoxetine treatment.
Fear of flying is a common specific phobia usually treated with behavior therapy; the possibility of pharmacologic treatment has received very little study. The authors report two cases in which successful fluoxetine treatment of major depressive disorder unexpectedly and unintentionally resolved a concomitant fear of flying that had preceded the depressive disorder.


The reactions of parents who think that a child is dying in a seizure--in their own words.
The goal of this work was to report the words parents use to describe a seizure in which they thought their child was dying, so as to increase the awareness of professionals with respect to the impact seizures may have on family life. An audit was carried out on 234 cases from a specialist epilepsy center and 35 cases from a tertiary referral outpatient clinic. Of the 54 parents who thought their child was dying in a seizure, in 45 cases the parents' exact words were recorded in the first person, and in 9 cases, their words were reported in the third person. Many parents took actions that illustrated their high degree of concern: 32 children were taken to hospital, 18 by ambulance, and 3 parents gave mouth-to-mouth resuscitation. Seizures often cause major concern in parents and many think that their child is dying. Prolonged seizures and limpness and/or cyanosis appear to be associated with this fear. Professionals can gain valuable insight into the impact of seizures on parents by asking them about their reactions to seeing their child having a seizure.


Modeling catalytic reaction mechanisms in glycoside hydrolases.
Modeling catalysis in carbohydrate-active enzymes is a daunting challenge because of the high flexibility and diversity of both enzymes and carbohydrates. Glycoside hydrolases (GHs) are an illustrative example, where conformational changes and subtle interactions have been shown to be critical for catalysis. GHs have pivotal roles in industry (e.g. biofuel or detergent production) and biomedicine (e.g. targets for cancer and diabetes), and thus, a huge effort is devoted to unveil their molecular mechanisms. Besides experimental techniques, computational methods have served to provide an in-depth understanding of GH mechanisms, capturing complex reaction coordinates and the conformational itineraries that substrates follow during the whole catalytic pathway, providing a framework that ultimately may assist the engineering of these enzymes and the design of new inhibitors.


Comparative study between iohexol and iopromide for aortofemoral arteriography.
A double-blind clinical trial was performed on 50 patients in order to compare omnipague (iohexol) and Ultravist-300 (iopromide) for peripheral arteriography. Both contrast media were well tolerated, but there was less body heat and pain in the Ultravist group. There were no idiosyncratic reactions or significant changes in blood pressure before and after the injection.


An investigation of the HMGR gene and IPI gene expression in black caraway (Bunium persicum).
Black caraway is of great importance for its terpene compounds. Many genes are involved in the biosynthesis of secondary metabolites in medicinal plants. For this study, black caraway seeds were collected from five different regions, i.e. [Isfahan; Kerman (Khabr); Semnan; Kerman (Sirch); and Hormozgan]. The black caraway seed oil was extracted and analyzed by means of the gas chromatography method. There was a negatively significant correlation (p ≤ 0.05) observed between cuminaldehyde and gammaterpinene compounds. 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and isopentenyl pyrophosphate isomerase (IPI) play an important role in the biosynthesis of secondary metabolites. Appropriate primers were designed for these genes based on the conserved regions in other plants. Amplified fragments were then sequenced. Blastn results indicated the similarity of the high RNA sequences between new sequences and other HMGR and IPI gene sequences in GenBank, and it also identified the HMGR and IPI gene sequences of B. persicum. A fragment of the HMGR gene with KJ143741 number was recorded in the gene bank. Quantitative PCR showed that the relative expression of two genes in different growth stages of B. persicum was significantly different between the germination stage and the multi-leaf stage, and also between the germination stage and the flowering stage (p < 0.05); however, there was no significant difference observed between the flowering stage and the multi-leaf stage. The results indicated that the expression of HMGR increased from the germination stage to the adult plant, and then it got stable until the flowering stage; in the same vein, the expression of IPI increased continuously from the germination stage to the flowering stage. The expression of HMGR and IPI genes occurred differently at the germination stage of five ecotypes. The Hormozgan ecotype showed the least expression rate.


Dynamic compliance of single perfused frog mesenteric capillaries and rat venules: a filtration coefficient correction.
The compliance of microvessels produces a potential source of error in the measurement of hydraulic conductance, regardless of which method is used. The compliance characteristics of rat and frog microvessels, in vivo, was studied using a two oil technique capable of measuring changes in radius of 0.01 micron at intervals of 0.02 seconds. Two water insoluble immiscible oils were micro-injected into occluded segments of the vessel. Since the volume of the oil remained constant the position of the interface at different times, following a sudden change in applied pressure, could be used to determine the time course of changes in vessel diameter. It was found that approximately 90% of the compliance response had occurred within 0.4 seconds of applying a step change in pressure. This compliance response coincides with the initial change in fluorescence used in the measurement of fluid with the initial change in fluorescence used in the measurement of fluid filtration by the densitometric technique.


Femoral nerve neuropathy following the psoas hitch procedure.
To present two cases of femoral nerve neuropathy following a psoas hitch procedure performed during Leadbetter-Politano ureteric reimplantation, and emphasize the importance of this rare complication. Two 2-year-old boys with complex urological histories underwent Leadbetter-Politano ureteric reimplantations with a psoas hitch using non-absorbable sutures independently in Turkey and the UK. Postoperatively both developed a femoral nerve palsy, which required re-exploration and removal of the sutures. In the first case the femoral nerve was not identified, but in the second case re-exploration revealed the femoral nerve stretched by the nearby suture. Both boys made a full recovery from their neuropraxia. These two cases illustrate the potential for nerve involvement with the psoas hitch manoeuvre, and the importance of early recognition of this complication. It is recommended that if a psoas hitch procedure is performed great care should be taken to identify the nerves, and if none are seen then the sutures are placed superficially into the belly of the muscle as the nerves may be sited deeper within it.


"It is more Important than food sometimes"; Meanings and Functions of Music in the Lives of Autistic Adults Through a hermeneutic-phenomenological Lense.
Subjective experiences of autistic adults remain under-researched, especially in the field of art. To learn more about their uses and functions of music, we interviewed 13 autistic adults and through a hermeneutic-phenomenological analysis found four overarching themes: Well-being, Identity and self-development, Connectedness, and Negative experiences. Findings show a broad and deep meaning of music in their lives, spanning from seemingly trivial functions such as making chores more enjoyable, to existential questions, such as choosing to stay alive. We discuss the often-overlooked negative effects or experiences of music, noting that positive and negative effects must always be addressed together if we are to use music to lower stress and support well-being.


Changes in IR band areas and band shifts during water adsorption to lecithin and ceramide.
Adsorption of water to a phospholipid (lecithin) and a ceramide were studied by IR microspectroscopy equipped with a humidity control system and quartz crystal microbalance (QCM). The water weight ratios increase up to 12.2 wt% for lecithin and 1.2 wt% for ceramide at RH ~80%, with linear correlations with infrared OH (+NH) band areas. For lecithin, the 1230 cm-1 band (PO2-) and the 1735 cm-1 band (CO) shift to lower wavenumbers, while the 1060 cm-1 band (PO2-, POC) shift to higher wavenumber with RH. Band areas of phosphates (1230 and 1060 cm-1) increase with RH showing positive relations with the band area of bound water. Bound water molecules with shorter H bonds might be bound to these phosphate groups. Band areas of aliphatic CHs are negatively correlated with the increasing adsorption of free water. Free water molecules with longer H bonds might interact loosely with aliphatic chains of lecithin. For ceramide, only the 1045 cm-1 band (CO) shows a small red shift at higher RHs than 60%, indicating adsorption of bound water to CO bonds. Amounts of water molecules adsorbed to ceramide are very limited due to few adsorption of free water.


Breast cancer stem cells: Multiple capacities in tumor metastasis.
Breast cancer is the leading cause of cancer death among women worldwide. Accumulating evidence indicates that the local recurrent and/or distant metastatic tumors, the major causes of lethality in the clinic, are related to the aggressive phenotype of a small fraction of cancer cells loosely termed as cancer stem cells (CSCs), tumor initiating cells (TICs), or cancer metastasis-initiating cells (CMICs). Breast cancer stem cells (BCSCs) are shown to exhibit unique growth abilities including self-renewal, differentiation potential, and resistance to most anti-cancer agents including chemo- and/or radiotherapy, all of which are believed to contribute to the development and overall aggressiveness of the recurrent or metastatic lesions. It is in the urgent need not only to further define the nature of heterogeneity in each tumor but also to characterize the precise mechanisms governing tumor-host cross-talk which is assumed to be initiated by BCSCs. In this review, we will focus on recently identified key factors, including the BCSCs among circulating tumor cells, interaction of BCSCs with the host, epithelial mesenchymal transition (EMT), tumor microenvironment, the intrinsic resistance due to HER2 expression, potential biomarkers of BCSCs and cancer cell immune signaling. We believe that new evidence coming from both bench and clinical research will help to develop more effective approaches to control or significantly reduce the aggressiveness of metastatic tumors.


The immunocytochemical localization of glutamate in the electrosensory system of the gymnotiform fish, Apteronotus leptorhynchus.
The first processing station of the electrosensory system is the medullary electrosensory lateral line lobe. The electrosensory lateral line lobe of gymnotiform fish is a laminated structure with three anatomically distinct excitatory inputs: primary afferents from electroreceptors, the direct feedback from n. praeminentialis and indirect feedback from n. praeminentialis via a cerebellar lobe--the eminentia granularis posterior. We utilized immunocytochemistry on L-R White-embedded semithin sections, and immunogold on thin sections to localize glutamate and aspartate in the electrosensory lateral line lobe, its afferent inputs and efferent targets. The excitatory inputs and the electrosensory lateral line lobe projection neurons are immunopositive for glutamate only, suggesting that glutamate rather than aspartate is the excitatory transmitter of the electrosensory system.