Airborne contamination of wounds in joint replacement operations: the relationship to sepsis rates.
During operations for total joint replacement done in operating rooms with conventional ventilation the mean air contamination varied considerably among the 15 hospitals studied. The range was from 51 to as many as 539 bacteria-carrying particles per cubic metre. When the data from all the hospitals were grouped according to the mean level of bacterial airborne contamination, including operations done in control and in ultraclean air, there was a good correlation between the air contamination and the joint sepsis rate. There was also a correlation between the mean values of air contamination and the numbers of bacteria isolated from wound wash-out samples; but the apparent efficiency of the sampling method varied a great deal among the hospitals carrying out this procedure. From this data it would seem that by far the largest proportion of bacteria found in the wound after the prosthesis had been inserted reached it by the airborne route. With the mean air contamination found in the control series, 164 bacteria-carrying particles per cubic metre, this proportion was as much as 95 per cent. The risk of joint sepsis varied widely among the 19 hospitals. The differences between the highest and lowest being probably as much as 20-fold. However, the effect of an ultraclean air environment was asimilar at all hospitals.


Cellular stress and glucocorticoid hormones protect L929 mouse fibroblasts from tumor necrosis factor alpha cytotoxicity.
Adaptive responses to the environment depend on the induction of the "stress response" in less differentiated organisms and cultured cells and the activation of the hypothalamic-pituitary-adrenal axis in animals and humans. This indicates that adrenal steroids and stress proteins play an important role in regulating cell survival in response to noxious stimuli. In an in vitro model, we analyzed the effects of either dexamethasone (DEX) treatment or environmental changes which can elicit a stress response, on the survival of cultured L-929 mouse fibroblasts exposed to the cytotoxic cytokine tumor necrosis factor alpha (TNF-alpha). DEX treatment produced a significant reduction in the apoptotic death of L-929 cells produced by TNF-alpha. Abrogation of the protective effect of DEX by actinomycin D and cycloheximide demonstrated that protection against TNF-alpha requires de novo synthesis of mRNA and proteins. The results were similar when L-929 cells were exposed to metabolic (serum starvation) or thermal (heat shock) stresses before TNF-alpha treatment. In both cases the stress process afforded significant protection against TNF-alpha cytotoxicity. Inhibition of mRNA and protein synthesis abrogated the protection exerted by stress (serum starvation) or produced massive death during the stress event (heat shock). The similarities in the protective activities of DEX and stress response and the reported interactions between heat shock proteins and glucocorticoid hormones suggest that stress proteins and glucocorticoids both belong to an ancient evolutionary pathway which controls cell survival.


From student to graduate: longitudinal changes in the qualities of nurses.
To examine the development of perceived qualities of nursing from student to graduate nurse over time. Researchers continue to explore student nurse and new graduate nurse attrition, particularly in the light of a looming crisis in nursing recruitment and retention. Qualities of nurses represent the job fit of nursing from student to graduate years. A prospective longitudinal design with a convenience sample was used for this study. Data were collected annually from 2009-2012 through the completion of a short on-line survey. The sample size of undergraduate nurses in year 1 was 676, with 527 in year 2, 339 in year 3 and 190 in year 4. Only 136 participants completed the survey each year forming the complete data set for analysis. Most qualities of nursing differed significantly across time with the qualities of Caring, Empathetic, Knowledge and Respectful demonstrating strong changes. Most declines in scores occurred on graduation. Caring, the central tenet of nursing increased during the student years and declined slightly on graduation. This unique longitudinal study of Australian nurses suggests that the clinical experience and theoretical grounding provided in our University programs, has resulted in an increasing cumulative effect in the third year supporting most qualities of nurses/nursing understood in year 1, that is, the career fit to perceptions, has been achieved. The decline in the 1(st) year of graduation, where the concept of workplace misfit is occurring, is where further nurse graduate support is urgently required.


End-stage renal disease: endocrine aspects of treatment.
End-stage renal disease (ESRD) is a deadly disease unless supportive treatment is administered in the form of haemodialysis, peritoneal dialysis or kidney transplantation. Although marked improvements have occurred in the efficiency of dialysis and in overall care, patients with ESRD still have poor long-term survival. The outcome is largely dependent on age, nutritional status, efficiency of dialysis and underlying reason for renal failure. As a consequence of renal failure, these patients experience a number of endocrine and metabolic disorders that may affect their well being and overall outcome. Disturbances in the somatotropic axis have been documented at several different levels, including an end-organ resistance to both growth hormone (GH) and insulin-like growth factor-I (IGF-I). A consequence seen in childhood is reduced growth velocity and short final height that may be overcome by long-term GH treatment, and it is possible that metabolism and nutritional status in adults with ESRD may be influenced by these abnormalities. Although a few small trials of GH treatment in adults with ESRD suggest that nutritional status may improve, long-term trials are needed to demonstrate other benefit of such treatment. This review will give a brief description of endocrine problems in adult patients with ESRD with a focus on the somatotropic axis, and it will review the experience reported in published trials of GH treatment in this patient group.


Copper exposure impairs intra- and extracellular acid-base regulation during hypercapnia in the fresh water rainbow trout (Oncorhynchus mykiss).
In order to evaluate the impact of water-borne copper on acid-base regulation in fresh water rainbow trout, chronically cannulated fish were exposed to copper (0.6 mg 1(-1)), hypercapnia (water PCO2 of 6 mmHg) or a combination of copper and hypercapnia, while a fourth untreated group served as the control. Blood samples obtained at 0 h, 4 h and 24 h were analysed for acid-base status, ion concentrations and respiratory parameters. Tissue samples from caudal skeletal muscle, liver and gill filaments were examined for intracellular acid-base status, ion- and water contents, and copper concentration. Exposure to copper alone elicited a small extracellular metabolic alkalosis, no changes in arterial PO2, and a minor decrease in plasma ion concentrations. Hypercapnia alone increased arterial PCO2 from approximately 2 mmHg to 7.2 mmHg, but the extracellular respiratory acidosis present at 4 h was almost completely compensated at 24 h due to an increase in plasma bicarbonate concentration [HCO3-] from 8.1 mM to 24.4 mM. Combined exposure to hypercapnia and copper resulted in a slightly larger acidosis at 4 h, and the fish failed to restore extracellular pH at 24 h, because plasma [HCO3-] only increased to 16.3 mM. Fish exposed to hypercapnia and copper also showed a delayed recovery of intracellular pH in skeletal muscle, compared to fish exposure to hypercapnia only. Thus, copper exposure impaired both extracellular and intracellular acid-base regulation during hypercapnia. When seen in connection with only minor effects of copper on osmoregulatory and respiratory parameters, the reduced ability to regulate acid-base suggests that acid-base regulation may be one of the most copper-sensitive branchial functions.


Comparison of 24-hour cardiovascular and autonomic function in paraplegia, tetraplegia, and control groups: implications for cardiovascular risk.
Fluctuations in 24-hour cardiovascular hemodynamics, specifically heart rate (HR) and blood pressure (BP), are thought to reflect autonomic nervous system (ANS) activity. Persons with spinal cord injury (SCI) represent a model of ANS dysfunction, which may affect 24-hour hemodynamics and predispose these individuals to increased cardiovascular disease risk. To determine 24-hour cardiovascular and ANS function among individuals with tetraplegia (n=20; TETRA: C4-C8), high paraplegia (n=10; HP: T2-T5), low paraplegia (n=9; LP: T7-T12), and non-SCI controls (n=10). Twenty-four-hour ANS function was assessed by time domain parameters of heart rate variability (HRV); the standard deviation of the 5-minute average R-R intervals (SDANN; milliseconds/ms), and the root-mean square of the standard deviation of the R-R intervals (rMSSD; ms). Subjects wore 24-hour ambulatory monitors to record HR, HRV, and BP. Mixed analysis of variance (ANOVA) revealed significantly lower 24-hour BP in the tetraplegic group; however, BP did not differ between the HP, LP, and control groups. Mixed ANOVA suggested significantly elevated 24-hour HR in the HP and LP groups compared to the TETRA and control groups (P<0.05); daytime HR was higher in both paraplegic groups compared to the TETRA and control groups (P<0.01) and nighttime HR was significantly elevated in the LP group compared to the TETRA and control groups (P<0.01). Twenty-four-hour SDANN was significantly increased in the HP group compared to the LP and TETRA groups (P<0.05) and rMSSD was significantly lower in the LP compared to the other three groups (P<0.05). Elevated 24-hour HR in persons with paraplegia, in concert with altered HRV dynamics, may impart significant adverse cardiovascular consequences, which are currently unappreciated.


[Clinico-endoscopic evaluation of the effectiveness of the treatment of uncomplicated gastroduodenal ulcers after local administration of solcoseryl and aevit].
The complex of conservative measures for noncomplicated peptic ulcers included curative endoscopy in 105 patients, in 61 of them Aevitum was used as an application with additional injections of Solcoseryl around the ulcers. In 44 patients Aevitum was used along with Solcoseryl which was injected into the submucous layer around the pathological focus of the stomach and duodenum. The results obtained gave a convincing evidence of advantages of interstitial administration of Aevitum allowing to considerably shorten time of treatment (up to 10-12 days) of noncomplicated ulcers of the stomach and duodenum.


The P1 family: a new class of nuclear mammalian proteins related to the yeast Mcm replication proteins.
Monospecific antibodies against an oligopeptide, conserved among the Mcm class of yeast replication proteins, were used to screen a human cDNA library. Eight of the isolated cDNA clones have the potential to code for sections of proteins with high sequence similarities to the yeast proteins Mcm3 and Cdc46 from Saccharomyces cerevisiae and Cdc21 from S. pombe. Our results establish a novel and highly conserved family of nuclear proteins in mammalian cells.


Enhanced adsorption selectivity of Au(III) over Cu(II) from acidic chloride solutions by chitosan/palm kernel fatty acid distillate/magnetite nanocomposites.
This work aimed to develop a modified chitosan adsorbent with enhanced adsorption selectivity for Au(III) over Cu(II) from acidic chloride solutions using low-cost and green raw materials. Various adsorbents, i.e., chitosan powder, chitosan microbeads, chitosan/palm kernel fatty acid distillate (PKFAD) microcomposites, magnetite nanoparticles, and chitosan/PKFAD/magnetite nanocomposites (CPMNs), were first evaluated for their ability to adsorb Au(III) and Cu(II) from single- and binary-metal solutions across different pH levels, followed by parametric analysis of Au(III) and Cu(II) adsorption from binary- and multi-metal solutions onto CPMNs, Au(III) desorption from Au(III)-loaded CPMNs, and reusability of CPMNs. Finally, Au(III)-loaded CPMNs were characterized with SEM-EDX, XRD, FTIR, and XPS to confirm the proposed adsorption mechanisms. Among all the adsorbents studied, CPMNs exhibited outstanding performance in adsorbing Au(III) from an equimolar binary Au(III)-Cu(II) solution, achieving the highest equilibrium adsorption capacity of 0.479 mmol/g (94.4 mg/g) without reaching saturation. Under optimal adsorption conditions of pH 3, 1 g/L CPMN dosage, and 90 min contact time, CPMNs adsorbed 96 % of Au(III) with a selectivity over Cu(II) exceeding 99 %. CPMNs demonstrated excellent reusability, maintaining over 80 % adsorption and desorption efficiencies for 5 cycles. The proposed adsorption mechanisms of CPMNs for Au(III) encompass electrostatic attraction, hydrogen bonding, solvation, and reduction.


A polymer/oil based nanovaccine as a single-dose immunization approach.
The recognized necessity for new antigen delivery carriers with the capacity to boost, modulate and prolong neutralizing immune responses prompted our approach, in which we describe a multifunctional nanocarrier consisting of an oily nanocontainer protected by a polymeric shell made of chitosan (CS), named CS nanocapsules (CSNC). The CS shell can associate the antigen on its surface, whereas the oily core might provide additional immunostimulating properties. In this first characterization of the system, we intended to study the influence of different antigen organizations on the nanocarrier's surface (using the recombinant hepatitis B surface antigen -rHBsAg- as a model antigen) on their long-term immunopotentiating effect, without any additional immunostimulant. Thus, two prototypes of antigen-loaded CSNC (CSNC+ and CSNC-), exhibiting similar particle size (200 nm) and high antigen association efficiency (>80%), were developed with different surface composition (polymer/antigen ratios) and surface charge (positive/negative, respectively). The biological evaluation of these nanovaccines evidenced the superiority of the CSNC+ as compared to CSNC- and alum-rHBsAg in terms of neutralizing antibody responses, following intramuscular vaccination. Moreover, a single dose of CSNC+ led to similar IgG levels to the positive control. The IgG1/IgG2a ratio suggested a mixed Th1/Th2 response elicited by CSNC+, in contrast to the typical Th2-biased response of alum. Finally, CSNC+ could be freeze-dried without altering its physicochemical properties and adjuvant effect in vivo. In conclusion, the evaluation of CSNC+ confirms its interesting features for enhancing, prolonging and modulating the type of immune response against subunit antigens, such as rHBsAg.


Complement resistance mechanisms of streptococci.
Group A streptococcus (GAS, Streptococcus pyogenes), group B streptococcus (GBS, Streptococcus agalactiae) and pneumococcus (Streptococcus pneumoniae) are all human pathogens that cause significant morbidity and mortality worldwide. These related species cause different spectra of infections spanning from trivial upper respiratory tract or skin infections to septic and severe diseases. In order to cause deep infections and survive in the human body the bacteria must evade the immune system. Complement is an important part of innate immunity both as an opsonizing and membrane destructing cascade and as an effector system of antibodies. In this review, we describe the complement resistance mechanisms of the three clinically most important streptococcal species, groups A and B streptococci and pneumococcus. The complement evasion mechanisms of these three species are analogous, yet different from one another. Several strains of all three species express molecules (M-proteins, Bac or beta, PspC) that acquire host fluid-phase complement regulators factor H or C4b binding protein to their surfaces. Groups A and B streptococci also secrete proteins and/or enzymes that inhibit the activation of the complement system or chemotaxis caused by the complement activation products. Even though a lot is known about the immune evasion by streptococci, the high morbidity and mortality associated with infections caused by streptococci and the need for efficient vaccines warrant further studies on the streptococcal molecules mediating complement resistance.


Structure-affinity relationship in the interactions of human organic anion transporter 1 with caffeine, theophylline, theobromine and their metabolites.
It is well known that human organic anion transporter 1 (hOAT1) transports many kinds of drugs, endogenous compounds, and toxins. However, little is known about the structure-affinity relationship. The aim of this study was to elucidate the structure-affinity relationship using a series of structurally related compounds that interact with hOAT1. Inhibitory effects of xanthine- and uric acid-related compounds on the transport of p-aminohippuric acid were examined using CHO-K1 cells stably expressing hOAT1. The order of potency for the inhibitory effects of xanthine-related compounds on PAH uptake was 1-methyl derivative>7-methyl derivative>3-methyl derivative falling dotsxanthine>1,3,7-trimethyl derivative (caffeine). The order of potency of the inhibition was 1,3,7-trimethyluric acid>1,3-dimethyluric acid>1,7-dimethyluric acid>1-methyluric acid>uric acid. A significant correlation between inhibitory potency and lipophilicity of the tested uric acid-related compounds was observed. The main determinant of the affinity of xanthine-related compounds is the position of the methyl group. On the other hand, lipophilicity is the main determinant of the affinity of uric acid-related compounds.


On the variability of chlorogenic acid concentration.
The plasticity of phenolic acid concentration in plants under various physiological stress situations raises questions concerning the ecological significance of such behavior. Helianthus annuus L. is remarkably plastic with regard to chlorogenic and ischlorogenic acid concentrations when subjected to nitrogen deficiency, NaCl-stress, short exposures to UV radiation or to combinations of these stresses. Stress due to NaCl has not previously been reported to produce this effect.I propose that chlorogenic acid is representative of a group of chemicals which have originated as regulators of various metabolic systems under stress and have subsequently acquired allelochemic properties against pathogens, herbivores, and competitors. If this hypothesis is correct then specific patterns of distribution with regard to concentrations of phenolic acids should be found. Some such predictions are discussed and means of distinguishing between various selective agents are explored. Generally, if natural environmental stress stimulates phenolic acid concentrations and hence more efficient growth patterns, then for several kinds of variables, higher concentrations should be found in more rather than less stressful habitats. If biological agents are significant agents of natural selection for this group of substances, such a pattern should not occur in some cases.


Unprotected stroke management in an undiagnosed case of Severe Acute Respiratory Syndrome Coronavirus 2 infection.
Coronavirus disease 19 (COVID-19) is an emerging infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this scenario, managing acute medical conditions, such as stroke, requires a timely treatment together with proper strategies that minimize the risk of infection spreading to health care workers and other patients. We report the case of a 79-year-old woman, who was admitted for a wake-up stroke due to occlusion of the left middle cerebral artery. She was treated outside the COVID-19-dedicated track of the hospital because she had no concomitant signs or symptoms suggestive of SARS-CoV-2 infection nor recent contact with other infected individuals. Post-mortem nasal and pharyngeal swab was positive for SARS-CoV-2 infection. We propose that hyperacute stroke patients should be tested for SARS-CoV-2 infection at admission and then managed as having COVID-19 until cleared by a negative result. We are aware that such measure results in some delay of the acute treatment of stroke, which could be minimal using well-exercised containment protocols.


Transfer of nitrazepam across the human placenta.
Six women from 14 to 17 weeks pregnant, and 12 woman from 36 to 40 weeks pregnant, were given nitrazepam 5 mg orally about 12 h before legal abortion by hysterotomy in the former group and elective caesarean section in the latter group. The concentration of nitrazepam was determined by gas-liquid chromatography. Binding to plasma proteins was evaluated by separation of the protein-free fraction by ultracentrifugation. In the first group (early pregnancy) the level of nitrazepam was found to be lower in the fetal than in the maternal circulation. The concentration in amniotic fluid was still lower. In the latter group (late pregnancy) the concentration both of unbound and total nitrazepam in maternal and fetal plasma were in equilibrium, which indicated an increase in transplancental transfer in late pregnancy. The percentage of unbound nitrazepam in both cases was 12%.


Partitioning of carbon an SO2-sulfur in a native grassland.
The seasonal assimilation and within-plant partitioning of 14CO2-carbon and 35SO2-sulfur in field plots of mixed-grass prairie was investigated, as was the dry deposition of 35SO2 onto surfaces of dead leaves, litter, and soil, and possible effects of continuous low-level SO2 fumigation on these processes. The proportion of total net-assimilated carbon found below-ground was 45% in May, 51% in July, and 17% in September. As the season progressed, greater proportions of assimilate were partitioned to 5-20 cm depths and less to the 0-5 cm depth. Rhizomes and crowns received greater proportions in late season. Significant fractions of total 34SO2-deposited sulfur were recovered on dead leaf surfaces as well as litter and soil, suggesting estimates of SO2 removal based on stomatal resistance alone are inadequate. Only 4% to 7% of total deposited sulfur was translocated belowground, with most going to 0-5 cm roots. In July much greater proportions of the total translocated SO2-sulfur were found in deeper depths than in September. On SO2-fumigated plots roots had lower total sulfur concentrations than controls. Furthermore, while on control plots total sulfur in roots at 5-20 cm increased from May to July and decreased from July to September, on fumigated plots there was a decrease followed by an increase suggesting that SO2 uptake by shoots interferes with the normal pattern of root sulfur uptake and redistribution within the plant. Continuous SO2 fumigation also seemed to stimulate root growth in July, possibly through a stimulation of photosynthesis.


B cell function in patients with chronic pancreatitis and its relation to exocrine pancreatic function.
Exocrine pancreatic function was evaluated by a Lundh meal test and a secretin-cholecystokinin test in 16 patients with chronic pancreatitis. B cell function was assessed by measuring the concentration of C-peptide after stimulation with oral glucose and intravenous glucagon. The Cc-peptide response to intravenous glucagon and oral glucose was closely correlated (r = 0.88, p less than 0.01). Plasma C-peptide after glucagon was significantly correlated to the post-prandial concentration of lipase (r = 0.72, p less than 0.001), amylase (r = 0.64, p less than 0.05) and to amylase output (r = 0.64, p less than 0.05). Eight out of nine patients treated with insulin had residual B cell function, but it diminished significantly with increasing duration of diabetes. We conclude that B cell function is correlated to pancreatic enzyme secretion and that patients with insulin-treated diabetes secondary to chronic pancreatitis have a residual insulin secretion similar to that of patients with Type 1 (insulin-dependent) diabetes.


Getting to the heart of cannabis health risks.
In this issue of Cell, Wei et al. show that the increased cardiovascular risks associated with cannabis use are mediated by proinflammatory cannabinoid 1 (CB1) receptor signaling, which can be ameliorated with the natural antioxidant agent genistein.


Novel brain reactive autoantibodies: prevalence in systemic lupus erythematosus and association with psychoses and seizures.
Autoantibodies can cause neuropsychiatric manifestations in lupus patients by altering the physiological function of neuronal cells. In this study, we identified Brain Reactive Autoantibodies (BRAAs) against murine neuronal membrane proteins (M.W. 27.5 and 29.5 kD) and found them correlating with psychosis and/or seizures in lupus patients. They were specific to neuronal membrane tissues of mammalian origin and are significantly associated with psychosis and/or seizures (p<0.0001). These membrane proteins mass spectrometry profiles did not match to any published protein sequences. These BRAAs may play important roles in the pathophysiology of neuropsychiatric lupus.


Feasibility study of volumetric modulated arc therapy for the treatment of retroperitoneal sarcomas.
Radiotherapy for retroperitoneal sarcomas remains controversial and a technical challenge considering the threshold of contiguous critical organs tolerance. We performed consecutive RapidArc dosimetric plans in preoperative or postoperative setting. A dosimetric study was carried out from six preoperative (group A) and four postoperative (group B) CT-scans, performed in 7 patients.Prescribed dose was 45 and 50 Gy for groups A and B, respectively. The planning target volume (PTV) was defined as the clinical target volume (CTV) plus 5 mm. The CTV encompassed the gross tumor volume (GTV) plus 10 mm or the tumoral bed. The dosimetric plans were optimized on a RapidArc Eclipse console using the progressive resolution algorithm, PRO version 8.8. Normalization method allowed the coverage of 99% of the PTV by 95% of the dose. Mean PTV were 2318.5 ± 2223.9 cc [range 348-6198 cc] and 698.3 ± 216.6 cc [range 463 -933 cc] for groups A and B, respectively. Plans were optimized for single arcs in group B and for single or two arcs in group A. The contralateral kidney volume receiving 5 Gy (V5Gy) was 21.5 ± 23.3% [range 0-55%] and 3.1 ± 2.6% [range 0-7.3%] for groups A and B, respectively. The mean dose received by 1% of the kidney (D1%) was 5.6 ± 2.4 Gy [range 3.6 -7.6 Gy] for group A and 5.4 ± 0.7 Gy [range 4.3-6 Gy] for group B. The volume of small bowel excluding the PTV (small bowel-PTV) that received 40 Gy and 30 Gy (V40Gy and V30Gy) in group A were 7.5 ± 4.4% [range 5.4-14.1%] and 18.5 ± 7.1% [range 10-30.4%], respectively.In group B, small bowel-PTV V40Gy and V30Gy were 4.7 ± 3.3% [range 3.3-8%] and 21.6 ± 7.5% [range 9.4-30%] respectively. In a second step, we treated two patients in the postoperative group. Treatment time delivery with one arc was 74 seconds. No severe acute toxicity was observed. RapidArc technology for retroperitoneal sarcomas showed acceptable dosimetric results in preoperative or postoperative clinical situation. From the first treated patients, acute tolerability was good to excellent.


SARS coronavirus: unusual lability of the nucleocapsid protein.
The severe acute respiratory syndrome (SARS) is a contagious disease that killed hundreds and sickened thousands of people worldwide between November 2002 and July 2003. The nucleocapsid (N) protein of the coronavirus responsible for this disease plays a critical role in viral assembly and maturation and is of particular interest because of its potential as an antiviral target or vaccine candidate. Refolding of SARS N-protein during production and purification showed the presence of two additional protein bands by SDS-PAGE. Mass spectroscopy (MALDI, SELDI, and LC/MS) confirmed that the bands are proteolytic products of N-protein and the cleavage sites are four SR motifs in the serine-arginine-rich region-sites not suggestive of any known protease. Furthermore, results of subsequent testing for contaminating protease(s) were negative: cleavage appears to be due to inherent instability and/or autolysis. The importance of N-protein proteolysis to viral life cycle and thus to possible treatment directions are discussed.


Comparative quantitative study of the hippocampal region of two closely related species of wild mice: interspecific and intraspecific variations in volumes of hippocampal components.
To investigate the structural changes in the hippocampal region (subiculum, Ammon's horn, and area dentata) associated with speciation, the volumes of homologous components of this region of the forebrain were compared in the two closely related murine rodent species, Apodemus flavicollis (yellow-necked wood mice) and A. sylvaticus (long-tailed field mice), and in two geographically separated groups of A. sylvaticus. With the exception of the mossy fiber zone, no significant differences were found in the relative sizes of the components of Ammon's horn. Significant interspecific differences were found in the deep subiculum, in the intermediate (medial perforant path) and deep (commissural-associational) zones of the molecular layer of the area dentata, and in the hilus. A significant intraspecific difference was found in the combined intermediate and superficial (medial and lateral perforant path) zone of the dentate molecular layer. Differences found in the relative size of the hilus and the mossy fiber zone of Ammon's horn were complementary in that the combined volumes of these zones, which are both terminal fields of dentate granule cells, did not differ in the species. This finding provides evidence that the distribution of the terminal field of a projection system can be altered while the size of the terminal field is maintained. Within the hippocampal region, components of Ammon's horn appear well suited for quantitative comparative studies that span taxonomic units beyond the species level. In agreement with previous quantitative studies, differences in the volumes of components of Ammon's horn found in species of different orders are more likely to reflect phylogenetic trends rather than changes resulting from specializations of the particular species used. This is not true for the subiculum and the components of the area dentata. Experimentally induced changes in the connectivity of the hippocampal region are discussed in terms of the structural changes which may be responsible for the quantitative differences observed between the two species studied here.


Xenografts Derived From Patients' Ascites Recapitulate the Gemcitabine Resistance Observed in Pancreatic Cancer Patients.
Most patient-derived pancreatic ductal adenocarcinoma (PDAC) xenografts have been established from surgical specimens of patients who have not received chemotherapy. However, xenografts have rarely been established from chemotherapy-resistant, advanced PDACs, because such cases are usually inoperable. The purpose of this study is to establish patient-derived xenografts using PDAC cells refractory to chemotherapy. Clinical PDAC cells obtained from ascites of patients who had received continuous chemotherapy were implanted into the flanks of immunocompromised mice. Growth and histological features of the xenografts with and without gemcitabine treatment were then analyzed. Ascites-derived PDAC cells were successfully expanded through serial xenograft passage without changes in histological appearance. While treatment with gemcitabine substantially inhibited the growth of all PDAC xenografts tested, the tumor volume gradually increased, and the tumors showed marked regrowth even under continued gemcitabine treatment. These findings are consistent with the actual clinical course of the corresponding patients for each xenograft. Ascites-derived xenograft models represent a valuable experimental system for testing the efficacy of currently available therapeutic compounds on chemotherapy-resistant PDAC cells and for elucidation of the mechanisms underlying chemotherapy resistance.


Avoiding the mental foramen during periapical surgery.
Many endodontists avoid surgical procedures in proximity to the mental foramen. Although a healthy respect for regional anatomy is important, this should not be at the expense of failing to offer the most appropriate endodontic care. When correct preoperative diagnostic and surgical techniques are used, serious negative sequelae are rarely encountered. Three simple steps used during presurgical diagnosis, flap design, and surgery are presented to increase the practitioner's confidence and safety while performing mandibular periapical surgery.


Risk Factors for Muscle Loss in Hemodialysis Patients with High Comorbidity.
With expanding kidney transplantation programs, remaining hemodialysis patients are more likely to have a high comorbidity burden and may therefore be more prone to lose muscle mass. Our aim was to analyze risk factors for muscle loss in hemodialysis patients with high comorbidity. Fifty-four chronic hemodialysis patients (Charlson Comorbidity Index 9.0 ± 3.4) were followed for 20 weeks using 4-weekly measurements of lean tissue mass, intracellular water, and body cell mass (proxies for muscle mass), handgrip strength (HGS), and biochemical parameters. Mixed models were used to analyze covariate effects on LTM. LTM (-6.4 kg, interquartile range [IQR] -8.1 to -4.8), HGS (-1.9 kg, IQR -3.1 to -0.7), intracellular water (-2.11 L, IQR -2.9 to -1.4) and body cell mass (-4.30 kg, IQR -5.9 to -2.9) decreased in all patients. Conversely, adipose tissue mass increased (4.5 kg, IQR 2.7 to 6.2), resulting in no significant change in body weight (-0.5 kg, IQR -1.0 to 0.1). Independent risk factors for LTM loss over time were male sex (-0.26 kg/week, 95% CI -0.33 to -0.19), C-reactive protein above median (-0.1 kg/week, 95% CI -0.2 to -0.001), and baseline lean tissue index ³10th percentile (-1.6 kg/week, 95% CI -2.1 to -1.0). Age, dialysis vintage, serum albumin, comorbidity index, and diabetes did not significantly affect LTM loss over time. In this cohort with high comorbidity, we found universal and prominent muscle loss, which was further accelerated by male sex and inflammation. Stable body weight may mask muscle loss because of concurrent fat gain. Our data emphasize the need to assess body composition in all hemodialysis patients and call for studies to analyze whether intervention with nutrition or exercise may curtail muscle loss in the most vulnerable hemodialysis patients.


Clinically and histologically silent Q fever endocarditis accidentally diagnosed by PCR.
A case of Q fever endocarditis was diagnosed in a patient with no sign of active endocarditis by performing PCR targeting eubacterial 16S rDNA on the resected mitral valve. The diagnosis was confirmed by detection of high levels of anti-Coxiella burnetti antibodies, positive immunohistologic analysis of the valve tissue with specific antibodies and culture of C. burnetti from the valve tissue. As this patient had an unexplained aggravation of valve dysfunction, we recommended routine serologic testing for C. burnetti to allow the diagnosis of Q fever endocarditis at a very early stage.


Mixture Analyses of Air-sampled Pollen Extracts Can Accurately Differentiate Pollen Taxa.
The daily pollen forecast provides crucial information for allergic patients to avoid exposure to specific pollen. Pollen counts are typically measured with air samplers and analyzed with microscopy by trained experts. In contrast, this study evaluated the effectiveness of identifying the component pollens using the metabolites extracted from an air-sampled pollen mixture. Ambient air-sampled pollen from Munich in 2016 and 2017 was visually identified from reference pollens and extracts were prepared. The extracts were lyophilized, rehydrated in optimal NMR buffers, and filtered to remove large proteins. NMR spectra were analyzed for pollen associated metabolites. Regression and decision-tree based algorithms using the concentration of metabolites, calculated from the NMR spectra outperformed algorithms using the NMR spectra themselves as input data for pollen identification. Categorical prediction algorithms trained for low, medium, high, and very high pollen count groups had accuracies of 74% for the tree, 82% for the grass, and 93% for the weed pollen count. Deep learning models using convolutional neural networks performed better than regression models using NMR spectral input, and were the overall best method in terms of relative error and classification accuracy (86% for tree, 89% for grass, and 93% for weed pollen count). This study demonstrates that NMR spectra of air-sampled pollen extracts can be used in an automated fashion to provide taxa and type-specific measures of the daily pollen count.


Blockade of metabotropic glutamate receptor 5 protects against DNA damage in a rotenone-induced Parkinson's disease model.
Glutamate excitotoxicity contributes to the development of Parkinson's disease (PD) and pharmacological blockade of metabotropic glutamate receptor 5 (mGluR5) has beneficial anti-akinetic effects in animal models of PD; however, the mechanism by which these antagonists alleviate PD symptoms is largely unknown. In our study, the effects of mGluR5 inhibition on DNA damage were investigated in a rotenone-induced model of PD. We first found that the selective mGluR5 antagonist, 2-methyl-6- (phenylethynyl) pyridine, prevented rotenone-induced DNA damage in MN9D dopaminergic neurons through a mechanism involving the downregulation of intracellular calcium release which was associated with a reduction in endoplasmic reticulum stress and reactive oxygen species (ROS)-related mitochondrial dysfunction. Interestingly, the ROS-related mitochondrial dysfunction was accompanied by an increase in expression of the antioxidant protein, Trx2. Treatment of cells with the calcium chelating agent 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or the ROS scavenger N-acetyl-L-cysteine, also reduced rotenone-induced DNA damage, while transfection of a dominant-negative form of Trx2 increased it. In addition, mGluR5 inhibition altered the expression profiles of proteins involved in DNA repair activity. Specifically, the expression of phosphorylated ERK (p-ERK) and CREB, as well as APE1 and Rad51 were elevated after rotenone stimulation and were subsequently downregulated following blockade of mGluR5. These findings were confirmed in vivo in a rotenone-induced rat model of PD. Inhibition of mGluR5 protected against neurotoxicity by mitigating oxidative stress-related DNA damage associated with 8-hydroxy-2'-deoxyguanosine production and also reduced p-ERK activity and Trx2 expression. These findings provide a novel link between mGluR5 and DNA damage in a model of PD, and reveal a potential mechanism by which mGluR5 mediates DNA damage in neurodegenerative diseases.


Administrators, boards, physicians must help change health system.
Change in the health care system is inevitable, and hospital administrators, boards, and medical staffs must learn not only to accept it but also to participate in effecting it. They must promote cooperation among hospitals, become more informed about the system's problems and pressures for change, and participate in activities of HSAs and other health-related groups that plan and influence change.


Glycolysis-based drug delivery nanosystems for therapeutic use in tumors and applications.
Tumor cells are able to use glycolysis to produce energy under hypoxic conditions, and even under aerobic conditions, they rely mainly on glycolysis for energy production, the Warburg effect. Conventional tumor therapeutic drugs are unidirectional, lacking in targeting and have limited therapeutic effect. The development of a large number of nanocarriers and targeted glycolysis for the treatment of tumors has been extensively investigated in order to improve the therapeutic efficacy. This paper reviews the research progress of nanocarriers based on targeting key glycolytic enzymes and related transporters, and combines nanocarrier systems with other therapeutic approaches to provide a new strategy for targeted glycolytic treatment of tumors, providing a theoretical reference for achieving efficient targeted treatment of tumors.


[Course and progression of children admitted before 4 years of age in a French child welfare center].
The purpose of this study was to determine the institutional trajectory and future of young children in child welfare. A catamnestic study - based on data from the child welfare office in Maine and Loire, France, from 1994 to 2001 - was conducted by a child psychiatrist and a psychologist. Medical, judicial, and educational data (development, health, pathways in child protection services) were collected and analyzed regarding the status of these children 15 years later, adding information gathered by interviewing the child welfare and foster family consultant. We included 128 children admitted to the child welfare office before 4 years of age. Admission to the child welfare system suffers from care delays (a mean of 13.1 months between the first child protection referral and placement) with an average entry age of 17 months and frequent cases of child abuse (e.g., seven Silverman syndrome cases). The physical and mental health status of these children was poor (poorly monitored pregnancies, prematurity, low birth weight). More than one third of the children had growth failure at admission, with catch-up in half of the cases. The average length of stay in the child welfare system was 13.2±4.6 years. At the end of the follow-up, there were specific measures to safeguard vulnerable adults: "young adult" (24 cases), "major protection" (eight cases) and "disabled living allowance" (nine cases). One hundred and sixteen children suffered from psychiatric disorders at entry and 98 at the end. The general functioning of children as assessed by the Children's Global Assessment Scale (CGAS) showed a statistically significant improvement. One out of two young adults showed problems integrating socially with chaotic pathways: many foster placements, unsuccessful return to the family, and academic failures. The clinical situations of children in the child welfare office and their long-term progression confirm the importance of this public health problem. Although the measures can greatly improve their physical and psychological recovery, with evidence of thriving, this remains limited: only a few of these children are well integrated socially and academically.


Muscular activity level during pedalling is not affected by crank inertial load.
The aim of the present study was to investigate the influence of gear ratio (GR) and thus crank inertial load (CIL), on the activity levels of lower limb muscles. Twelve competitive cyclists performed three randomised trials with their own bicycle equipped with a SRM crankset and mounted on an Axiom ergometer. The power output ( approximately 80% of maximal aerobic power) and the pedalling cadence were kept constant for each subject across all trials but three different GR (low, medium and high) were indirectly obtained for each trial by altering the electromagnetic brake of the ergometer. The low, medium and high GR (mean +/- SD) resulted in CIL of 44 +/- 3.7, 84 +/- 6.5 and 152 +/- 17.9 kg.m(2), respectively. Muscular activity levels of the gluteus maximus (GM), the vastus medialis (VM), the vastus lateralis (VL), the rectus femoris (RF), the medial hamstrings (MHAM), the gastrocnemius (GAS) and the soleus (SOL) muscles were quantified and analysed by mean root mean square (RMS(mean)). The muscular activity levels of the measured lower limb muscles were not significantly affected when the CIL was increased approximately four fold. This suggests that muscular activity levels measured on different cycling ergometers (with different GR and flywheel inertia) can be compared among each other, as they are not influenced by CIL.


An overactive mind: Investigating racing thoughts in ADHD, hypomania and comorbid ADHD and bipolar disorder via verbal fluency tasks.
Adults with Attention Deficit Hyperactivity Disorder (ADHD) often report mental restlessness akin to racing thoughts found in hypomanic and mixed episodes of bipolar disorder (BD). Past research has suggested that racing thoughts in BD can be tackled via process-oriented verbal fluency measures. In ADHD, it is still unknown whether racing thoughts is due to comorbid BD, and its neuropsychological underpinnings remain to be investigated. To this aim, this study investigates process-oriented verbal fluency measures in adults with ADHD compared to adults with BD in a hypomanic episode, adults with ADHD + BD and healthy controls. Three verbal fluency tasks, i.e., the free, the letter and the semantic conditions, were administered to 37 adults with ADHD, 25 adults with BD in a hypomanic episode, 22 adults with comorbid ADHD + BD (euthymia) and 31 healthy subjects. Word production, clustering, and switching were analysed in verbal fluency tasks. Adults with ADHD, ADHD + BD and hypomania showed increased switches in the free verbal fluency task, compared to healthy controls, despite equivalent number of words produced. Unlike the ADHD and ADHD + BD groups, phonological clustering measures in the semantic task were increased in the hypomanic group compared to healthy controls. BD groups were under medication. Semantic overactivation, reflected by increased switches in the unconstrained verbal fluency task, appear to be a common mechanism involved in racing thoughts in both ADHD and BD. By contrast, unusual sounds-based associations are specific of hypomanic speech and might contribute to the distinction trait and state racing thoughts.


P53 mutations in colorectal cancer from northern Iran: Relationships with site of tumor origin, microsatellite instability and K-ras mutations.
CRC-associated P53 mutations have not been studied extensively in non-Western countries at relatively low CRC risk. We examined, for the first time, 196 paraffin-embedded CRC cases from Northern Iran for mutations in P53 exons 5-8 using PCR-direct sequencing. P53 status and mutation site/type were correlated with nuclear protein accumulation, clinicopathologic variables and data on K-ras mutations and high-level microsatellite instability (MSI-H). We detected 96 P53 mutations in 87 (44.4%) cases and protein accumulation in 84 cases (42.8%). P53 mutations correlated directly with stage and inversely with MSI-H. Distal CRCs were more frequently mutated at major CpG hotspot codons [248 (8/66, 12.1%), 175 (7/66, 10.6%), and 245 (7/66, 10.6%)], while in proximal tumors codon 213, emerged as most frequently mutated (5/28, 17.9% vs. 3/66, 4.5%, P = 0.048). Transitions at CpGs, the most common mutation type, were more frequent in non-mucinous (25% vs. 10.4% in mucinous, P = 0.032), and distal CRC (27% vs. 12.5% in proximal, P = 0.02), and correlated with K-ras transversions. Transitions at non-CpGs, second most common P53 mutation, were more frequent in proximal tumors (15.6% vs. 4.7% in distal, P = 0.01), and correlated with K-ras transitions and MSI-H. Overall frequency and types of mutations and correlations with P53 accumulation, stage and MSI-H were as reported for non-Iranian patients. However P53 mutation site/type and correlations between P53 and K-ras mutation types differed between proximal and distal CRC. The codon 213 P53 mutation that recurred in proximal CRC was previously reported as frequent in esophageal cancer from Northern Iran.


Studies on trimethylsilyl derivatives of 1-alk-1enyl-2-acyl glycerols by gas-liquid chromatography mass spectrometry.
Trimethylsilyl derivatives of 1-hexadec-1-enyl-2-decanoyl and -2-octadecenoylglycerols and 1-octadec-1-enyl-2-decanoyl and -2-octadecenoylglycerols were subjected to analysis by a gas chromatograph mass spectrometer system. The characteristic ions are as follows: (1) ions at [M-15]+ and [M-90]+- are observed, 2 or 16 mass units less than those of the corresponding 1-alkyl-2-acyl or 1,2-diacyl derivatives; (2) the base peak is m/e 129; (3) the ion at [M-R1CH=CHO]+ or [R2COO+130]+ is present in high abundance; (4) ions at m/e 311 and 339, characteristic of trimethylsilyl derivatives of 1-hexadec-1-enyl and 1-octadec-1-enylglycerols, are due to [R1CH-CHO-1)+73]+, also confirmed from studies on the perdeuteriotrimethylsilyl derivatives; (5) several ions based on one acyl group, [R2CO]+, [R2CO+74]+ and [R2COO+74]+ are also observed.


Complete nucleotide sequence of a new virus, peach chlorotic leaf spot virus, isolated from flat peach in China.
Contigs with sequence homologies to apple chlorotic leaf spot virus (ACLSV) were identified by high-throughput sequencing analysis in three peach samples. Complete genomic sequences of RP19-1 and RP19-2 of the virus consisted of 7,466 and 7,465 nucleotides (nts), respectively, excluding the poly (A) tails. They shared the highest identity with Ta Tao 5, but lower than 70% of sequence similarity with other ACLSV isolates. Furthermore, phylogenetic analysis revealed that these two isolates clustered with Ta Tao 5, which is distinct from other ACLSV isolates. According to the criteria for species demarcation within the genus Trichovirus, these two isolates as well as Ta Tao 5 should be recognized as a new virus species, tentatively named "Peach chlorotic leaf spot virus".


Structural studies of the Escherichia coli O-antigen 6.
The structure of the Escherichia coli O-antigen 6 has been investigated using n.m.r. spectroscopy, methylation analysis, and various specific degradations. It is concluded that the O-antigen is composed of pentasaccharide repeating-units having the following structure. (Formula: see text)


Influence of face mask design on bag-valve-mask ventilation performance: a randomized simulation study.
Different face mask designs can influence bag-valve-mask (BVM) ventilation performance during resuscitation. We compared a single-use, air-cushioned face mask (AM) with a reusable silicone face mask (SM) for quality of BVM ventilation on a manikin simulating cardiac arrest. Thirty-two physicians were recruited, and a prospective, randomized, crossover observational study was conducted after an American Heart Association-accredited basic life support provider course and standardized practice time were completed. Participants performed 12 cycles of BVM ventilation with both the AM and SM on a SmartMan lung simulator. Mean tidal volume was significantly higher in ventilations performed using the AM vs. the SM (548 ± 159 ml vs. 439 ± 163 ml, P < 0.01). In addition, the proportion of low-volume ventilation was significantly lower with the AM than the SM [6/12 (2-11) vs. 9/12 (5-12), P = 0.03]. Bag-valve-AM ventilation volume was not affected by the physical characteristics of the rescuers, except for sex. In contrast, bag-valve-SM ventilation volume was affected by most of the characteristics tested, including sex, height, weight, hand width, hand length, and grip power. The AM seems to be a more efficient face mask than the SM at delivering sufficient ventilation volumes. The performance of the AM did not seem to be associated with the physical characteristics of the rescuers, whereas that of the SM was affected by these factors. The SM may not be an appropriate face mask for performing one-person BVM ventilation during resuscitation for rescuers who are smaller in stature, have a smaller hand size, or have weaker grip power.


Improved differential diagnosis of breast masses on ultrasonographic images with a computer-aided diagnosis scheme for determining histological classifications.
A computer-aided diagnosis (CAD) scheme for determining histological classifications of breast masses is expected to be useful for clinicians in making a differential diagnosis. The purpose of this study was to evaluate the usefulness of using the CAD scheme on ultrasonographic images. The database consisted of 390 breast ultrasonographic images with masses. Three experienced clinicians independently provided subjective ratings on the likelihood of malignancy for each of the 390 masses. Fifty benign masses (25 cysts and 25 fibroadenomas) and 50 malignant masses (25 noninvasive ductal carcinomas and 25 invasive ductal carcinomas) were selected as unknown cases for an observer study based on a stratified randomization method with the ratings. The likelihood of the histological classification in each unknown case was evaluated by the CAD scheme with image features that clinicians commonly use for describing masses. In the observer study, seven observers provided their confidence levels regarding the malignancy of the unknown case before and after viewing the likelihood of the histological classification. The usefulness of the CAD scheme was evaluated with a multireader multicase receiver operating characteristic (ROC) analysis. The areas under the ROC curves (AUCs) for all observers were improved by use of the CAD scheme. The average AUC increased from 0.716 without to 0.864 with the CAD scheme (P = .006). The presentation of the likelihood of the histological classification evaluated by the CAD scheme improved the clinicians' performance and therefore would be useful in making a differential diagnosis of masses on ultrasonographic images.


Effects of Pelleting and Long-Term High-Temperature Stabilization on Vitamin Retention in Swine Feed.
The objective of this study was to study the effect of pelleting and long-term high-temperature stabilization on the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in swine feed. Piglet diets (diet 1 and 3) were pelleted after conditioning at 83 °C for 120 s, and were high-temperature stabilized at 90 °C for 8.5 min after pelleting; the finishing pig diets (diet 2, 4, and 5) were pelleted after conditioning at 82 °C for 90 s, and were high-temperature stabilized at 85 °C for 9 min after pelleting; the samples were obtained before condition, after condition, after pelleting, and after cooling. The contents of vitamin A and vitamin E in diets 1−5 and vitamin B2, and vitamin B6 in diets 3−5 were detected. The results showed that: (1) the conditioning process had no significant effect on the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in all experimental diets (p > 0.05); (2) the pelleting process and high-temperature stabilization process after pelleting had different degrees of influence on vitamins, among which the stabilization process had a more significant effect on the retention of vitamins. After pelleting and long-term high-temperature stabilization, the retention of vitamin A, vitamin E, and B2, and vitamin B6 were 68.8−77.3%, 56.9−90.1%, 63.8−70.3%, and 60.1−67.0%, respectively. In the process of pelleting and long-term high-temperature stabilization, the retention of vitamin A, vitamin E, vitamin B2, and vitamin B6 in the feed were significantly reduced (p < 0.05). Therefore, vitamin loss during high temperature and over a long period of time is worth considering, and vitamins must be over-supplemented.


Comparison of the forward and sideways locomotor patterns in children with Cerebral Palsy.
Switching locomotion direction is a common task in daily life, and it has been studied extensively in healthy people. Little is known, however, about the locomotor adjustments involved in changing locomotion direction from forward (FW) to sideways (SW) in children with cerebral palsy (CP). The importance of testing the ability of children with CP in this task lies in the assessment of flexible, adaptable adjustments of locomotion as a function of the environmental context. On the one hand, the ability of a child to cope with novel task requirements may provide prognostic cues as to the chances of modifying the gait adaptively. On the other hand, challenging the child with the novel task may represent a useful rehabilitation tool to improve the locomotor performance. SW is an asymmetrical locomotor task and requires a differential control of right and left limb muscles. Here, we report the results of a cross-sectional study comparing FW and SW in 27 children with CP (17 diplegic, 10 hemiplegic, 2-10 years) and 18 age-matched typically developing (TD) children. We analyzed gait kinematics, joint moments, EMG activity of 12 pairs of bilateral muscles, and muscle modules evaluated by factorization of EMG signals. Task performance in several children with CP differed drastically from that of TD children. Only 2/3 of children with CP met the primary outcome, i.e. they succeeded to step sideways, and they often demonstrated attempts to step forward. They tended to rotate their trunk FW, cross one leg over the other, flex the knee and hip. Moreover, in contrast to TD children, children with CP often exhibited similar motor modules for FW and SW. Overall, the results reflect developmental deficits in the control of gait, bilateral coordination and adjustment of basic motor modules in children with CP. We suggest that the sideways (along with the backward) style of locomotion represents a novel rehabilitation protocol that challenges the child to cope with novel contextual requirements.


Segmentation by classification: A novel and reliable approach for semi-automatic selection of HIV/SIV envelope spikes.
We describe a semiautomated approach to segment Env spikes from the membrane envelope of Simian Immunodeficiency Virus visualized by cryoelectron tomography of frozen-hydrated specimens. Multivariate data analysis is applied to a large set of overlapping subvolumes extracted semiautomatically from the viral envelope and does not utilize a template of the target structure. The major manual step used in the method involves determination of six points that define an ellipsoid approximating the virion shape. The approach is robust to departures of the actual virion from this starting ellipsoid. A point cage of sufficient density is generated to ensure that any spike-like protein is identified multiple times. Subsequently translational alignment of class averages to a cylindrical reference on a curved surface separates subvolumes with spikes from those without. Spike containing subvolumes identified multiple times are removed by proximity analysis. Slightly different procedures segment spikes in the equatorial and the polar regions. Once all spikes are segmented, further alignment of class averages using separately the polar and spin angles produces recognizable spike images. Our approach localized 96% of the equatorial spikes and 85% of all spikes identified manually; it identifies a significant number of additional spikes missed by manual selection. Two types of spike shapes were segmented, one with near 3-fold symmetry resembling the conventional spike, the other had a T-shape resembling the spike structure obtained when antibodies such as PG9 bind to HIV Env. The approach should be applicable to segmentation of any protein spikes extending from a cellular or virion envelope.


The role of the current waveform in mitigating passivation and enhancing electrocoagulation performance: A critical review.
Electrocoagulation (EC) can be an efficient alternative to existing water and wastewater treatment methods due to its eco-friendly nature, low footprint, and facile operation. However, the electrodes applied in the EC process suffer from passivation or fouling, an issue resulting from the buildup of poorly conducting materials on the electrode surface. Indeed, such passivation gives rise to various operational problems and restricts the practical implementation of EC on a large scale. Therefore, it has been suggested that using pulsed direct current (PDC), alternating pulse current (APC), and sinusoidal alternating current (AC) waveforms in EC as alternatives to conventional direct current (DC) can help mitigate passivation and alleviate its associated detrimental effects. This paper presents a critical review of the impact of the current waveform on the EC process towards the capabilities of the PDC, APC, and AC waveforms in de-passivation and performance enhancement while comparing them to the conventional DC. Additionally, current waveform parameters influencing the surface passivation of electrodes and process efficiency are elaborately discussed. Meanwhile, the performance of the EC process is evaluated under different current waveforms based on pollutant removal efficiency, energy consumption, electrode usage, sludge production, and operating cost. The proper current waveforms for treating various water and wastewater matrices are also explained. Finally, concluding remarks and outlooks for future research are provided.


Assessing the relationship between food insecurity and lifestyle behaviors among university students: a comparative study between Lebanon and Germany.
Food insecurity is a common public health problem in both developed and developing countries. This study aimed to profile food insecurity among university students in a developed country with stable economic circumstances (Germany) and a developing Mediterranean country undergoing a severe economic and financial crisis (Lebanon) and examine the associations between food insecurity and lifestyle behaviors (i.e., physical activity, sleep, and adherence to a healthy eating pattern, such as the Mediterranean diet), stress, and financial well-being. This online cross-sectional study was conducted between September 2021 and March 2022. Subjects were recruited through social media platforms (Facebook, WhatsApp, Instagram, and personal email) and in-class announcements by several university professors of various majors and from different universities in Lebanon and Germany. The final sample included 547 participants (197 from Lebanon and 350 from Germany). Our findings showed a higher food insecurity rate in Lebanon compared with Germany (59% versus 33%). In the bivariate analysis, food insecurity was associated with insomnia (r = 0.230; p < 0.001) and stress (r = 0.225; p = 0.001); German university students had higher physical activity (p < 0.001), better diet quality (p < 0.001), and lower adherence to the Mediterranean diet (p < 0001) than Lebanese students. In the multivariable analyses, more stress was related to insomnia (B = 0.178; p < 0.001), while financial well-being was not associated with any of the lifestyle behaviors. Physical activity, insomnia, and Mediterranean diet adherence were not associated with the country or food insecurity (p > 0.05); however, living in Germany was associated with better diet quality (B = -7.85; p < 0.001). The high prevalence of food insecurity reported in this study is alarming, particularly among Lebanese students; German students had better diet quality and higher physical activity but worse adherence to the Mediterranean diet. Moreover, food insecurity was also associated with worse sleep and stress. Further studies are necessary to assess the role of food insecurity as a mediating factor between sociodemographic characteristics and lifestyle behaviors.


Translocation of protein kinase C to subcellular fractions of human neutrophils.
The subcellular localization of protein kinase C in unstimulated human neutrophils and neutrophils stimulated by phorbol-myristate-acetate (PMA), 1-oleoyl-2-acetyl-rac-glycerol (OAG), and ionomycin was investigated in subcellular fractions obtained by nitrogen cavitation and Percoll density gradient centrifugation. Protein kinase C was found to be localized mainly in the cytosol in unstimulated cells, whereas significant translocation to fractions containing the plasma membrane was observed after stimulation by PMA, OAG, and ionomycin. At the same time, phospholipid-insensitive protein kinase activity appeared in the cytosol and the plasma membrane fractions. To determine whether binding of protein kinase C occurred to the plasma membrane or to intracellular membranes that had translocated to the plasma membrane, we investigated the ability of isolated azurophil, specific and secretory granules, and plasma membrane vesicles to bind protein kinase C in response to addition of PMA and OAG. Only fractions containing plasma membranes and secretory granules were able to bind protein kinase C. The observation explains the selective activation of plasma membrane structures by protein kinase C.


Is delayed ischemic preconditioning as effective on running performance during a 5km time trial as acute IPC?
Ischemic preconditioning (IPC) may enhance exercise performance. Cardioprotective effects of IPC are known to re-occur 24h after the stimulus. Whether the delayed effect of IPC has similar effects as IPC on exercise performance is unknown. Examine whether IPC applied 24h (24-IPC) before exercise is equally effective as IPC in improving exercise performance. Randomized, cross-over study METHODS: 12 healthy participants were randomly exposed to SHAM-session, IPC or 24-IPC before a self-paced 5km running trial on a treadmill. Subjects were blinded for time, speed and heart rate. Furthermore, heart rate, BORG, and the local tissue saturation index were measured during exercise, while lactate levels were determined after running. Using a regression model, we explored whether these parameters predicted the change in running time after IPC and 24-IPC. We found no differences in finish time after IPC (SHAM: 1400±105s, IPC: 1381±112s, 24-IPC: 1385±113s; p=0.30). However, we observed a significant positive relation between the change in finish time after IPC and 24-IPC (p=0.016; r=0.677). Using stepwise linear regression, a lower post-exercise blood lactate level after IPC or 24-IPC was significantly related to an improvement in finish time (R2=0.47, β=-0.687, p=0.007). Although no significant effect of IPC or 24-IPC on exercise performance was found, individual finish time after IPC and 24-IPC were strongly correlated. Therefore, our data suggest that, at the individual level, the effects of 24-IPC are closely related to the effects of IPC.


Ectomychorrizal DB: a symbiotic association database.
Ectomycorrhizal (ECM) fungal species, a "Symbiotic" relationship between tress and fungi in forest has a great ecological and economic importance. Here is an attempt to describe database named "EctomycorrhizalDB", addressing ECM diversity of Central Himalaya (Kumaun region), with special emphasis on their characterization, physical properties and morphological features along with specifications. This database would help the scientific community to draw a better understanding of the environmental factors that affects species diversity. The database is available for free at http://www.kubic.nic.in/ectomychorhiza.


A comparison of BeadChip and WGS genotyping outputs using partial validation by sanger sequencing.
Head-to-head comparison of BeadChip and WGS/WES genotyping techniques for their precision is far from straightforward. A tool for validation of high-throughput genotyping calls such as Sanger sequencing is neither scalable nor practical for large-scale DNA processing. Here we report a cross-validation analysis of genotyping calls obtained via Illumina GSA BeadChip and WGS (Illumina HiSeq X Ten) techniques. When compared to each other, the average precision and accuracy of BeadChip and WGS genotyping techniques exceeded 0.991 and 0.997, respectively. The average fraction of discordant variants for both platforms was found to be 0.639%. A sliding window approach was utilized to explore genomic regions not exceeding 500 bp encompassing a maximal amount of discordant variants for further validation by Sanger sequencing. Notably, 12 variants out of 26 located within eight identified regions were consistently discordant in related calls made by WGS and BeadChip. When Sanger sequenced, a total of 16 of these genotypes were successfully resolved, indicating that a precision of WGS and BeadChip genotyping for this genotype subset was at 0.81 and 0.5, respectively, with accuracy values of 0.87 and 0.61. We conclude that WGS genotype calling exhibits higher overall precision within the selected variety of discordantly genotyped variants, though the amount of validated variants remained insufficient.


Determination of ochratoxin A in beer marketed in Spain by liquid chromatography with fluorescence detection using lead hydroxyacetate as a clean-up agent.
A new sample treatment for liquid chromatographic analysis of ochratoxin A (OTA) in beer is proposed. Degassed beer is mixed with lead hydroxyacetate, which precipitates some bulk components but does not remove OTA. The precipitate is separated and the acidified liquid is extracted with chloroform. The solvent is evaporated and the residue is dissolved in mobile phase (acetonitrile-water, 40:60, v/v; acidified at pH 3.0 with phosphoric acid) and separated by liquid chromatography using fluorescence detection. The limit of detection was 0.005 ng/ml. The average recovery rate and the average RSD of recovery in the spiking level range 0.01-0.5 ng/ml were 95.5% and about 5%, respectively. The method is cheaper that other alternative ones using immunoaffinity columns or other solid-phase extraction cleanup:The separation was optimised with regard to composition and flow of the mobile phase and no interference from the matrix was found. The method was applied to 88 samples of beer (domestic and imported) marketed in Spain. OTA was detected in 82.9% of them. The range for positive samples was 0.007-0.204 ng of OTA/ml.


The efficacy of fluazuron in the management of blowfly strike in sheep.
The blowflies Lucilia spp. and Chryomya spp. feature prominently as a leading cause of myiasis and production losses in sheep in South Africa. With chemical means of preventing and treating blowfly strike being a common manner of control, new molecules or formulations effective against blowfly strike are needed as resistance becomes more of a problem. For this study we evaluated the potential of topical application of fluazuron on sheep as an additional means to control blowflies. When pure fluazuron was applied to raw meat at the recommended dose for cattle, significant effects on adult fly development were observed. However when applied topically to post-mortally collected sheep pelts at the same dose rate, no significant effects were observed on larval repellence or larval development with fly development progressing as expected. A subsequent pharmacokinetic study showed virtually no systemic absorption together with significant wool binding. Despite demonstrating that fluazuron was effective against blowfly larvae, wool binding of the molecule preclude its use.


In-use testing of extemporaneously prepared suspensions of second generation non-nucleoside reversed transcriptase inhibitors in support of phase I clinical studies.
DPC 961 and 963 are two of a series of prospective second generation non-nucleoside reverse transcriptase inhibitors (NNRTIs) being considered for the treatment of HIV infections. The 'powder in a bottle' approach was used for drug administration for Phase I clinical studies. This new approach consists of compounding the active drug as a suspension or solution at the clinical site immediately before dosing. Prior to clinical use, studies were conducted to determine the compatibility of the drugs with the suspending agent, the recovery of the drugs using the administration procedure and the dissolution profile of the suspensions. The stability of the DPC 961 and 963 in the dosing formulation was followed over a 24-h period using a stability indicating HPLC method. In addition, the dissolution profiles of the suspensions were established for future comparison with solid dosage forms. Although the two drugs have very similar chemical structures, they clearly exhibited different behaviors in liquid/liquid extraction and dissolution experiments. These differences could be related to the physical characteristics of the powders, such as particle size and surface area. The results of the in-use testing of the suspension showed adequate recovery of the drugs from the bottle at two drug levels. The stability of DPC 961 and 963 in the suspending agent was sufficient for constitution and administration of the suspensions at the clinical site.


Regulation of mouse preimplantation development: inhibitory effect of the calmodulin antagonist W-7 on the first cleavage.
The calmodulin antagonist W-7 inhibits cleavage of 1-cell mouse embryos in a concentration-dependent manner. This inhibition is likely to be specific for a calmodulin-mediated process, since the less active congener W-5 does not inhibit cleavage when used at concentrations of W-7 that do. Concentrations of W-7 that inhibit cleavage and do not inhibit either the uptake or incorporation of [35S]methionine do inhibit [3H]thymidine incorporation; similar concentrations of W-5 do not inhibit [3H]thymidine incorporation. Consistent with W-7's ability to inhibit cleavage by inhibiting DNA synthesis is that addition of W-7 at later times that correspond with exit from S phase results in cleavage to the 2-cell stage. Although W-7 does inhibit cleavage of 1-cell embryos, it does not inhibit transcriptional activation, which occurs in the 2-cell embryo and is characterized by the synthesis of a group of proteins of Mr = 70,000. Results of these experiments suggest a role for calmodulin in the first cell cycle of the mouse embryo and provide another example in which zygotic gene activation is not dependent on progression through the first cell cycle.


A randomized clinical trial to compare the efficacy of erythromycin, ampicillin and tetracycline for the treatment of cholera in children.
To compare the clinical outcome of treatment of cholera in children with ampicillin, erythromycin or tetracycline, a double-'blind' randomized four-cell trial was carried out in Bangladesh. Ampicillin was chosen as additional therapy for acute respiratory tract infection, present in many subjects with diarrhoea. One hundred and eighty-four children aged 1-5 years who were not wasted, with diarrhoea of duration < 48 h, signs of some or severe dehydration, dark-field stool microscopy demonstrating Vibrio cholerae, and a baseline purging rate > 4 mL/kg/h over 6 h were enrolled in the study. Ampicillin, tetracycline, erythromycin or placebo were given orally every 6 h for 3 d. After 3 d of antibiotic treatment, diarrhoeal stool volume was significantly reduced in all antibiotic groups, with mean volumes per kg body weight as follows: tetracycline, 318 mL (SEM = 50), ampicillin, 335 mL (SEM = 30); erythromycin, 323 mL (SEM = 25); placebo, 498 mL (SEM = 37). Compared to tetracycline, the clinical recovery rates by 96 h were 75% with placebo, 91.3% with ampicillin, and 95.7% with eythromycin. Compared to tetracycline, the total mean times to recovery were increased by 66% with placebo (P < 0.001), 25% with ampicillin (P < 0.017), and 9% with erythromycin (P = 0.37). These results indicated comparable clinical efficacy of tetracycline, ampicillin and erythromycin. We therefore recommend that, unless V. cholerae is resistant, ampicillin should be used as a cost-effective alternative to erythromycin for paediatric cholera, especially in children with concomitant acute respiratory infection.


Effects of 5'-N-ethylcarboxamideadenosine (NECA) on erythropoietin production.
The present studies were undertaken to assess the effects of 5'-N-ethylcarboxamideadenosine (NECA), an adenosine analogue, on erythropoietin (Epo) production. NECA (0.05 and 0.1 mumol/kg i.v.) produced significant increases in serum Epo levels (368.8 +/- 56.1 and 384.6 +/- 45.9 mU/ml, respectively) in exhypoxic polycythemic mice after a four hour exposure to hypoxia when compared with hypoxia controls (133.2 +/- 18.2 mU/ml). The hypoxic kidney Epo levels were 46.4 +/- 13.4 mU/kg kidney which were significantly higher than normoxic kidney Ep levels (< 1.24 mU/kg kidney). Theophylline (20 mg/kg i.p.), an adenosine receptor antagonist, significantly inhibited the stimulatory effects of NECA on serum Epo levels. In vitro cultures of an Epo producing hepatocellular carcinoma (Hep3B) cell line with NECA (> or = 10(-6) M) for 20 hours under hypoxic conditions (1% O2) produced significant increases in medium levels of Epo when compared with hypoxia controls. Hepatocellular carcinoma cells treated with NECA at a concentration range of 10(-7) M to 5 x 10(-5) M for one hour in a hypoxic atmosphere also had significantly higher cAMP levels than that of hypoxia controls. Scatchard analyses of [3H]NECA binding to membrane preparations of hepatocellular carcinoma cells showed low affinity binding sites with a dissociation-constant (Kd) of 0.44 microM and a binding capacity of 863 fmol/mg protein. These findings suggest that the increase in Epo production in response to NECA under hypoxic conditions can be attributed, at least in part, to stimulation of adenosine A2 receptors which is coupled to adenylyl cyclase activation.


Erratum to "Reproductive performance of early- and late-calving dairy cows artificially inseminated after ovulation synchronization and estrous resynchronization or artificially inseminated after observed estrus" (JDS Commun. 2:80-85).
[This corrects the article DOI: 10.3168/jdsc.2020-0035.].


Gardner's syndrome with duodenal adenomas, gastric adenomyoma and thyroid papillary--follicular adenocarcinoma.
A case of a woman with Gardner's syndrome, originally manifested by multiple adenomatous polyps of the colon, is presented. She underwent subtotal colectomy at 14 years of age. Over the next 15 years she had a composite odontoma, an impacted supernumerary tooth, two epidermal inclusion cysts, multicentric paillary-follicular adenocarcinoma of the thyroid, two tubulovillous adenomas of the duodenum in which argyrophilic cells were a prominent feature, and an adenomyoma of the gastric antrum. The presence of all of these lesions in one patient expands the spectrum of lesions seen with Gardner's syndrome and supports the concept of a generalized abnormality of growth regulation as the cause of the syndrome.


The importance of animal behavior for ecosystem services.
Animal behavior plays a critical role in the delivery of ecosystem services, yet the study of animal behavior and ecosystem services rarely intersect. The study of behavior-mediated ecosystem services should be prioritized, focusing on the conditions that allow these critical behaviors to persist and adapt to global change.


Personal-level exposure to environmental temperature is a superior predictor of endothelial-dependent vasodilatation than outdoor-ambient level.
Environmental temperatures influence cardiovascular physiology. However, the majority of time is spent indoors, making outdoor-ambient temperatures inaccurate estimates of true exposures encountered by most individuals. We evaluated in 50 healthy adults the associations between previous 7-day outdoor-ambient (four occasions) and prior 24-hour personal-level (two occasions) environmental temperature exposures with blood pressure, heart rate variability, sleep parameters, and endothelial-dependent vasodilatation (brachial flow-mediated dilatation [FMD]) using generalized estimating equations. Participants (34 females; age, 32.1 ± 9.6 years) had normal blood pressures (107.8 ± 13.3/70.2 ± 9.4 mm Hg), FMD (7.4 ± 2.8%), as well as sleep and heart rate variability parameters. Mean 7-day outdoor-ambient (4.6 ± 9.7°C) differed from personal-level temperature exposures (22.0 ± 3.0°C). Colder outdoor-ambient temperatures (per -10°C) over the previous 1-6 days (rolling averages) were associated with decreases in FMD: -0.57% (95% confidence interval [CI]: -1.14% to 0.01%, P = .055) to -0.62% (95% CI: -1.07% to -0.18%, P = .006). However, a 10°C decrease in personal-level temperature during the prior 24 hours was associated with a greater decrement in FMD: -2.44% (95% CI: -4.74% to -0.13%, P = .038). Both were also linearly related to FMD during all seasons and without a threshold temperature. Other end points were not significantly related to either temperature level in this study. Short-term exposures to colder environmental temperatures reduced endothelial-dependent vasodilatation, supporting the epidemiologic associations with heightened cardiovascular risk. We show here for the first time that temperature exposures characterized at the personal level may be more robust predictors of endothelial function than outdoor-ambient levels.


Peripherally administered sera antibodies recognizing amyloid-β oligomers mitigate Alzheimer's disease-like pathology and cognitive decline in aged 3× Tg-AD mice.
Active and passive immunotherapy targeting amyloid-β (Aβ) may be the most promising strategy to prevent or treat Alzheimer's disease (AD). Previously, immunization with the recombinant 6Aβ15-T antigen generated robust anti-Aβ serum antibodies that strongly recognized Aβ42 oligomers in different mice, markedly reduced the amyloid burden, and improved behavioral performance of immunized older AD mice. Here, we further determined that these anti-6Aβ15-T serum antibodies from different strains of mice displayed anti-Aβ antibody responses against the same epitopes in the Aβ1-15 region. Peripheral administration of anti-6Aβ15-T serum antibodies was also effective to mitigate AD-like pathology and cognitive decline in aged 3× Tg-AD mice. Specifically, the levels of Aβ and tau in the brains of 3× Tg-AD mice were significantly reduced after passive immunotherapy, which seemed necessary or beneficial to ameliorate memory impairment. In addition, our results showed that this immunotherapy also prevented presynaptic dynamin 1 degradation, which might help to further protect synaptic functions and allow functional recovery of cognition. Moreover, immunization with 6Aβ15-T in rabbits induced a similar antibody response as that in mice, and the rabbit serum antibodies reacted strongly with Aβ42 oligomers and inhibited oligomer-mediated neurotoxicity. We concluded that passive immunization with Aβ42 oligomer conformation-sensitive anti-6Aβ15-T serum antibodies is effective in providing potentially therapeutic effects in aged 3× Tg-AD mice by reducing Aβ and tau.


Genotoxicity of epoxy resin hardeners in the hepatocyte primary culture/DNA repair test.
The genotoxicity of 9 chemicals used as epoxy resin hardeners was examined in the DNA repair test with rat hepatocytes. DNA repair synthesis was elicited by 7 chemicals, i.e., 4-aminodiphenyl ether, 4,4-diaminodiphenyl ether, 3,4,4'-triaminodiphenyl ether, 3,3'-dichloro-4,4'-diaminodiphenyl ether, 1,3-phenylenedi-4-aminophenyl ether, 4,4'-diaminodiphenyl methane and 4,4'-methylene-bis(2-chloroaniline). The positive results obtained with 4 epoxy resin hardeners of unknown carcinogenicity, i.e., 4-amino-diphenyl ether, 3,4,4'-triaminodiphenyl ether, 3,3'-dichloro-4,4'-diaminodiphenyl ether and 1,3-phenylene-di-4-aminophenyl ether suggest that they may be carcinogens. The genotoxicity of 1,4-phenylene-di-4-aminophenyl ether, of unknown carcinogenicity, and 4,4'-diaminodiphenyl sulfone, for which there is no sound proof of carcinogenicity, was not confirmed in the DNA repair test. The result with 4,4'-diaminodiphenyl sulfone was in agreement with its lack of mutagenicity in Salmonella typhimurium.


Euglena gracilis and Its Aqueous Extract Constructed With Chitosan-Hyaluronic Acid Hydrogel Facilitate Cutaneous Wound Healing in Mice Without Inducing Excessive Inflammatory Response.
Naturally occurring compounds isolated from the microalga Euglena gracilis, such as polysaccharide paramylon, exhibit antimicrobial, anti-viral, antitumor, and anti-inflammatory activities. Whether live E. gracilis cells and its aqueous extract accelerate burn wound healing remains to be investigated. In this study, live E. gracilis cells and its aqueous extract were mixed with chitosan-hyaluronic acid hydrogel (CS/HA) to form cell + CS/HA and extract + CS/HA, which were then smeared onto the deeply burned skin of mice. The efficacy of these mixtures in accelerating wound healing was assessed through wound size reduction measurement, histological and immunofluorescence analyses, and serum pro-inflammatory cytokine level (INF-γ, IL-1β, and IL-6) determination. The live E. gracilis cells and its aqueous extract were found to facilitate wound healing by enhancing re-epithelization and reducing fibroplasia without stimulating excessive inflammatory response. In conclusion, live E. gracilis cells and its aqueous extract can be potentially used to treat cutaneous wounds.


4-Fluoro-2-({[(2R)-1-hy-droxy-1,1,3-tri-phenyl-propan-2-yl]imino}-meth-yl)phenol.
The title compound, C28H24FNO2, crystallizes in the ortho-rhom-bic space group P212121. A hydrogen-bonding network between the tertiary alcohol group and the fluoro substituent results in [010] chains in the solid state.


Pharmacological modulation of the micturition pattern in normal and cyclophosphamide pre-treated conscious rats.
In the current study, we wanted to assess the influence of muscarinic receptors, nitric oxide and purinoceptors on the micturition pattern of conscious normal and cyclophosphamide (CYP) pre-treated rats. The micturition parameters were assessed using a metabolic cage. Rats were pre-treated with either saline or CYP, to induce cystitis, followed by treatment with either the muscarinic M1/M3/M5 receptor antagonist 4-diphenylacetoxy-N-methylpiperidine (4-DAMP), the nitric oxide synthase blocker N(ω)-nitro-L-arginine methyl (L-NAME), the P2 purinoceptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) or a combination of 4-DAMP with PPADS or L-NAME. Voiding volumes per micturition event were significantly lower in CYP pre-treated than in saline pre-treated rats. Neither 4-DAMP nor L-NAME had any effect in the normal rats, whereas PPADS reduced the micturition volume per event. In CYP pre-treated rats, 4-DAMP and L-NAME significantly increased voiding volumes per event and micturition frequency, respectively. 4-DAMP dose-dependently reduced the differences in micturition activity between saline and CYP pre-treated rats. We show that cystitis changes the urodynamics in conscious rats and that this change seems to depend on the production of NO and on altered muscarinic receptor effects. The altered muscarinic receptor responses are likely to per se involve NO-mediated mechanisms.


The Atypical Kinesin KIF26A Facilitates Termination of Nociceptive Responses by Sequestering Focal Adhesion Kinase.
Kinesin superfamily proteins (KIFs) are molecular motors that typically alter the subcellular localization of their cargos. However, the atypical kinesin KIF26A does not serve as a motor but can bind microtubules and affect cellular signaling cascades. Here, we show that KIF26A maintains intracellular calcium homeostasis and negatively regulates nociceptive sensation. Kif26a-/- mice exhibit intense and prolonged nociceptive responses. In their primary sensory neurons, excessive inhibitory phosphorylation of plasma membrane Ca2+ ATPase (PMCA) mediated by focal adhesion kinase (FAK) rendered the Ca transients resistant to termination, and the peripheral axonal outgrowth was significantly enhanced. Upstream, KIF26A is directly associated with a FERM domain of FAK and antagonizes FAK function in integrin-Src family kinase (SFK)-FAK signaling, possibly through steric hindrance and localization to cytoplasmic microtubules.


A sensitive flow-batch system for on board determination of ultra-trace ammonium in seawater: Method development and shipboard application.
Combining fluorescence detection with flow analysis and solid phase extraction (SPE), a highly sensitive and automatic flow system for measurement of ultra-trace ammonium in open ocean water was established. Determination was based on fluorescence detection of a typical product of o-phthaldialdehyde and ammonium. In this study, the fluorescence reaction product could be efficiently extracted onto an SPE cartridge (HLB, hydrophilic-lipophilic balance). The extracted fluorescence compounds were rapidly eluted with ethanol and directed into a flow cell for fluorescence detection. Compared with the common used fluorescence method, the proposed one offered the benefits of improved sensitivity, reduced reagent consumption, negligible salinity effect and lower cost. Experimental parameters were optimized using a univariate experimental design. Calibration curves, ranging from 1.67 to 300nM, were obtained with different reaction times. The recoveries were between 89.5 and 96.5%, and the detection limits in land-based and shipboard laboratories were 0.7 and 1.2nM, respectively. The relative standard deviation was 3.5% (n=5) for an aged seawater sample spiked with 20nM ammonium. Compared with the analytical results obtained using the indophenol blue method coupled to a long-path liquid waveguide capillary cell, the proposed method showed good agreement. The method had been applied on board during a South China Sea cruise in August 2012. A vertical profile of ammonium in the South East Asia Time-Series (SEATS, 18° N, 116° E) station was produced. The distribution of ammonium in the surface seawater of the Qiongdong upwelling in South China Sea is also presented.


Correction: Identification of Potential Novel Biomarkers and Signaling Pathways Related to Otitis Media Induced by Diesel Exhaust Particles Using Transcriptomic Analysis in an In Vivo System.
[This corrects the article DOI: 10.1371/journal.pone.0166044.].


The control of cytosolic Ca2+ concentration: studies of high affinity Ca2+ transport in permeabilized acini of rat submandibular glands.
Active Ca2+ transport was studied in acini that had been permeabilized by incubation in buffer nominally free of Ca2+ in order to avoid any contribution to measured Ca2+ uptake by elements of the plasma membrane. In most experiments, ruthenium red was used to inhibit mitochondrial Ca2+ uptake. Non-mitochondrial Ca2+ transport was greatest at pH 7.0-7.5 and required Mg2+-ATP at concentrations typical of the cytosol (K0.5 = 1.36 +/- 0.53 mM). Other substrates were much less effective than ATP. Ca2+ uptake was stimulated by Ca2+ at concentrations near those measured in intact cells (K0.5 = 0.43 +/- 0.17 microM). Hill coefficients at subsaturating concentrations of Ca2+ (1.08 +/- 0.27) and Mg2+-ATP (0.81 +/- 0.22) indicated that cooperative interactions are not characteristic of the major cationic regulators of Ca2+ transporting activity. Na+ did not release Ca2+ from acinar mitochondria, but consistently reduced non-mitochondrial Ca2+ uptake by about 20% as compared to uptake in the presence of an equimolar K+ concentration. The properties of non-mitochondrial Ca2+ uptake in permeabilized acini are similar to those of high affinity Ca2+ uptake which, in broken cell preparations, has been found distributed in parallel with elements of the endoplasmic reticulum. The Ca2+-sequestering properties of a non-mitochondrial organelle in permeabilized submandibular gland acini are those expected of a principal regulator of cytosolic Ca2+ and could account for the ionized Ca2+ concentration measured in resting salivary acinar cells.


Circular dichroism of the peripheral chlorophylls in photosystem II reaction centers revealed by electrochemical oxidation.
Visible absorption spectra and circular dichroism (CD) of the red absorption band of isolated photosystem II reaction centers were measured at room temperature during progressive bleaching by electrochemical oxidation, in comparison with aerobic photochemical destruction, and with anaerobic photooxidation in the presence of the artificial electron acceptor silicomolybdate. Initially, selective bleaching of peripheral chlorophylls absorbing at 672 nm was obtained by electrochemical oxidation at +0.9 V, whereas little selectivity was observed at higher potentials. Illumination in the presence of silicomolybdate did not cause a bleaching but a spectral broadening of the 672-nm band was observed, apparently in response to the oxidation of carotene. The 672-nm absorption band is shown to exhibit a positive CD, which accounts for the 674-nm shoulder in CD spectra at low temperature. The origin of this CD is discussed in view of the observation that all CD disappears with the 680-nm absorption band during aerobic photodestruction.


Puberty.
Puberty consists of interrelated biological changes and occurs at a time when the individual also encounters cognitive and social changes. In this article we examine our current knowledge regarding the onset of puberty, the sequence and timing of puberty, and medical issues and physiologic changes that may arise with pubertal maturation. Figures included demonstrate the sexual maturation stages and interrelationships between the various parameters of puberty.


Identification of magnetic nanoparticles for combined positioning and lentiviral transduction of endothelial cells.
The combination of magnetic nanoparticles (MNPs) with a magnetic field is a powerful approach to enable cell positioning and/or local gene therapy. Because physical requirements for MNPs differ between these two applications we have explored whether the use of different MNPs can provide site-specific positioning combined with efficient viral transduction of endothelial cells (ECs). A variety of MNPs was screened for magnetic cell labeling and lentivirus binding. Then two different MNPs were chosen and their combined application was evaluated regarding EC magnetization and transduction efficiency. The combined use of PEI-Mag2 and NDT-Mag1 particles provided both efficient lentiviral transduction and high magnetic responsiveness of ECs that could be even retained within the vascular wall under flow conditions. The use of these MNPs did not affect biological characteristics of ECs like surface marker expression and vascular network formation. Importantly, with this method we could achieve an efficient functional overexpression of endothelial nitric oxide synthase in ECs. The application of two different MNPs provides optimal results for magnetic labeling of ECs in combination with viral transduction. This novel approach could be very useful for targeted gene therapy ex vivo and site-specific cell replacement in the vascular system.


First assessment of atmospheric pollution by trace elements and particulate matter after a severe collapse of a tailings dam, Minas Gerais, Brazil: An insight into biomonitoring with Tillandsia usneoides and a public health dataset.
In this study, samples of bromeliad Tillandsia usneoides (n = 70) were transplanted and exposed for 15 and 45 days in 35 outdoor residential areas in Brumadinho (Minas Gerais state, Brazil) after one of the most severe mining dam collapses in the world. Trace elements aluminum (Al), arsenic (As), chromium (Cr), copper (Cu), iron (Fe), mercury (Hg), manganese (Mn), nickel (Ni) and zinc (Zn) were quantified by atomic absorption spectrometry. Scanning electron microscope generated surface images of T. usneoides fragments and particulate matter (PM2.5, PM10 and PM > 10). Aluminum, Fe and Mn stood out from the other elements reflecting the regional geological background. Median concentrations in mg kg-1 increased (p < 0.05) between 15 and 45 days for Cr (0.75), Cu (1.23), Fe (474) and Mn (38.1), while Hg (0.18) was higher at 15 days. The exposed-to-control ratio revealed that As and Hg increased 18.1 and 9.4-fold, respectively, not showing a pattern associated only with the most impacted sites. The PM analysis points to a possible influence of the prevailing west wind on the increase of total particles, PM2.5 and PM10 in transplant sites located to the east. Brazilian public health dataset revealed increase in cases of some cardiovascular and respiratory diseases/symptoms in Brumadinho in the year of the dam collapse (1.38 cases per 1000 inhabitants), while Belo Horizonte capital and its metropolitan region recorded 0.97 and 0.37 cases, respectively. Although many studies have been carried out to assess the consequences of the tailings dam failure, until now atmospheric pollution had not yet been evaluated. Furthermore, based on our exploratory analysis of human health dataset, epidemiological studies are required to verify possible risk factors associated with the increase in hospital admissions in the study area.


Primary Plasma Cell Leukemia: A Retrospective Study of a Rare Disease From Tertiary Cancer Centre From India.
Plasma cell leukemia (PCL) is an aggressive rare leukemic variant of multiple myeloma (MM). We aim to present 4 years data on clinical profile and treatment outcomes of Primary PCL (PPCL) patients treated at tertiary care cancer centre from Northern India. To analyse response and safety profile of a PPCL with or without stem cell transplantation. Retrospectively reviewed and analysed PPCL patient's data at our centre from January-2013 to June-2017. Total 11 PPCL patients diagnosed among 240 MM patients during study period. Eight were males. Only 10 patients were started on treatment. Four (n = 4/10) patients underwent stem cell transplantation. Overall response rate was 70% (n = 7). Eleven culture positive bacterial infections (bloodstream = 2, urinary tract = 3; pulmonary = 6) were recorded. Four patients had fungal infections. One patient had Herpes Zoster infection. Relapse rate of entire cohort was 50% (n = 5). Median PFS and OS of entire cohort was 11 months (95% confidence interval 6.3-15.6) and 21 months (95% C.I. 1-49.8) respectively. The estimated PFS and OS at 1 year of transplanted versus nontransplanted patients were 71% + 24% versus 0% (P = 0.96) and 71% + 24% versus 15% + 19% (P = 0.234) respectively. Treatment with PIs + IMAs followed by transplants (single/double) might improve depth and duration of remission and OS. Patients should be treated with indefinite maintenance therapy to control disease.


Plant size, geitonogamy and seed set in Ipomopsis aggregata.
We used powdered fluorescent dyes to estimate receipt of self vs. outcross pollen in the self-incompatible species Ipomopsis aggregata (Polemoniaceae). Flowers on small and large plants received equal amounts of outcross pollen, whereas flowers on large plants received more self pollen, so the proportion of self pollen delivered through geitonogamy increased with plant size. In natural populations emasculation of all flowers on a plant raised average seed set per flower from 5.19 to 6.99 and also raised fruit set, though not significantly. From these results one expects a negative correlation between plant size and seeds per flower. The opposite trend was observed in a sample of plants in the field, suggesting that deleterious effects of geitonogamy on female fecundity in large plants can be overruled by other factors such as size-related fruit or seed abortion. Results are discussed in relation to the evolution of gynodioecy.