New insights into eutrophication management: Importance of temperature and water residence time.
Eutrophication and harmful cyanobacterial blooms threaten water resources all over the world. There is a great controversy about controlling only phosphorus or controlling both nitrogen and phosphorus in the management of lake eutrophication. The primary argument against the dual nutrients control of eutrophication is that nitrogen fixation can compensate the nitrogen deficits. Thus, it is of great necessary to study the factors that can significantly affect the nitrogen fixation. Due to the difference of climate and human influence, the water quality of different lakes (such as water temperature, N:P ratio and water residence time) is also quite different. Numerous studies have reported that the low N:P ratio can intensify the nitrogen fixation capacities. However, the effects of temperature and water residence time on the nitrogen fixation remain unclear. Thus, 30 shallows freshwater lakes in the eastern plain of China were selected to measure dissolved N2 and Ar concentrations through N2: Ar method using a membrane inlet mass spectrometer to quantify the nitrogen fixation capacities and investigate whether the temperature and water residence time have a great impact on nitrogen fixation. The results have shown that the short lake water residence time can severely inhibit the nitrogen fixation capacities through inhibiting the growth of nitrogen-fixing cyanobacteria, changing the N:P ratio and resuspending the solids from sediments. Similarly, lakes with low water temperature also have a low nitrogen fixation capacity, suggesting that controlling nitrogen in such lakes is feasible if the growth of cyanobacteria is limited by nitrogen.


Unfractionated human marrow cell cryopreservation using dimethylsulfoxide and hydroxyethyl starch.
Unfractionated bone marrow (BM) cells were cryopreserved in 1- to 2-ml aliquots using a mixture containing both 5% dimethylsulfoxide (DMSO) and 6% hydroxyethyl starch (HES) in an attempt to increase the viable cell yield and reduce the clumping after thawing, observed when 10% DMSO is used alone. Samples thawed after storage for 6 months in the vapor phase of liquid nitrogen, were assayed. Compared to prefreeze values, there was both a greater number of cells that excluded Trypan Blue (50 +/- 12 vs 28 +/- 12%, P less than .01) and a greater CFU-C Recovery (110 +/- 20 vs 89 +/- 35%, P less than .02) for cells in the DMSO/HES mixture, compared to those in 10% DMSO alone. No macroscopic clumping of the thawed cells was observed for those cryopreserved in the mixture in contrast to those in DMSO alone. Freezing was done without a rate-controlled freezing apparatus by simply placing the samples initially into a -80 degrees C freezer, and then later into a liquid nitrogen freezer. Additional samples stored in the DMSO/HES mixture were kept at only -80 degrees C, and when thawed 12 to 16 months later also gave an excellent CFU-C recovery (105 +/- 39% of prefreeze). The DMSO/HES mixture allows for a simplified BM cryopreservation technique that not only assures excellent recovery of CFU-Cs and eliminates clumping upon thawing, but also does not require either the use of a rate-controlled freezer or liquid nitrogen temperatures for storage up to a year.


Comparison of Intermittent versus Continuous Superior Venal Caval Oxygen Saturation Monitoring in Early Goal Directed Therapy in Septic Shock: A Systematic Review.
Early goal directed therapy (EGDT) is a bundle of care (monitoring ScvO 2 and lactate along with clinical parameters and instituting therapy) that has shown to improve outcomes in patients with septic shock. We conducted a systematic review of clinical trials and observational studies to compare intermittent versus continuous monitoring of ScvO 2 . We did major database searches till August 2020. Hospitalized children (>2 months age) and adults with septic shock were included. The intervention was "intermittent ScvO 2 monitoring," and the comparator was "continuous ScvO 2 monitoring." The primary outcome is "all-cause mortality." Of 564 citations, 3 studies ( n = 541) including both children and adults were included in the analysis. There was no significant difference in the "overall/all-cause mortality" (two randomized controlled trials; 258 participants) between the "intermittent" and "continuous" ScvO 2 monitoring groups (relative risk [RR]: 1.00; 95% confidence interval [CI]: 0.8-1.24). However, a single observational study (283 participants) showed a significant increase in mortality in the intermittent group (RR: 1.46; 95% CI: 1.03-2.05). The GRADE evidence generated for "overall/all-cause mortality" was of "moderate certainty." To conclude, the present meta-analysis did not find any significant difference between "intermittent" and "continuous" ScvO 2 monitoring in patients with septic shock.


A live human B-cell activator operating in isolation of other cellular influences.
Using a reversed haemolytic plaque assay, we demonstrate that live but not inactivated Epstein-Barr (EB) virus can induce human blood lymphocytes to polyclonal Ig (IgM and IgG) secretion in vitro. Unlike pokeweed mitogen this B-cell-activating effect takes place in relative absence of T cells. EB virus thus promises to be a tool for study of B cell function isolated from other cellular influences, although its mechanism of activation appears to be different from that of previously known B cell activators.


Genomic typing of hepatitis C viruses present in China.
Hepatitis C virus (HCV) genomic clones were obtained from the serum of Chinese HCV carriers using a polymerase chain reaction-based approach. Consensus sequences were derived from (1) the structural region (nt 1-1543) for one carrier, (2) the hypervariable region V (nt 1156-1233) from four carriers and (3) region V3 from four carriers. Region V3, located in the nonstructural domain NS5 (nt 7066-7137), has been previously shown to be a particularly good marker for the genomic typing of HCV isolates [Inchauspe et al., Proc. Natl. Acad. Sci. USA 88 (1991) 10292-10296]. Comparison of these sequences with sequences from geographically distinct HCV isolates indicates that Chinese HCV strains are closely related to, though distinguishable from, Japanese prototype strains. One amino acid motif, GGAA, located in region V, was found to be conserved only among Chinese isolates. This may define a new subgroup among HCV isolates.


Multiple γ-secretase product peptides are coordinately increased in concentration in the cerebrospinal fluid of a subpopulation of sporadic Alzheimer's disease subjects.
Alcadeinα (Alcα) is a neuronal membrane protein that colocalizes with the Alzheimer's amyloid-β precursor protein (APP). Successive cleavage of APP by β- and γ-secretases generates the aggregatable amyloid-β peptide (Aβ), while cleavage of APP or Alcα by α- and γ-secretases generates non-aggregatable p3 or p3-Alcα peptides. Aβ and p3-Alcα can be recovered from human cerebrospinal fluid (CSF). We have previously reported alternative processing of APP and Alcα in the CSF of some patients with sporadic mild cognitive impairment (MCI) and AD (SAD). Using the sandwich enzyme-linked immunosorbent assay (ELISA) system that detects total p3-Alcα, we determined levels of total p3-Alcα in CSF from subjects in one of four diagnostic categories (elderly controls, MCI, SAD, or other neurological disease) derived from three independent cohorts. Levels of Aβ40 correlated with levels of total p3-Alcα in all cohorts. We confirm that Aβ40 is the most abundant Aβ species, and we propose a model in which CSF p3-Alcα can serve as a either (1) a nonaggregatable surrogate marker for γ-secretase activity; (2) as a marker for clearance of transmembrane domain peptides derived from integral protein catabolism; or (3) both. We propose the specification of an MCI/SAD endophenotype characterized by co-elevation of levels of both CSF p3-Alcα and Aβ40, and we propose that subjects in this category might be especially responsive to therapeutics aimed at modulation of γ-secretase function and/or transmembrane domain peptide clearance. These peptides may also be used to monitor the efficacy of therapeutics that target these steps in Aβ metabolism.


The association of body mass index with complications and functional outcomes after surgery for closed ankle fractures.
This study assessed the association of classes of body mass index in kg/m2 (classified as normal weight 18.5 kg/m2 to 24.9 kg/m2, overweight 25.0 kg/m2 to 29.9 kg/m2, and obese ≥ 30.0 kg/m2) with short-term complications and functional outcomes three to six years post-operatively for closed ankle fractures. We performed a historical cohort study with chart review of 1011 patients who were treated for ankle fractures by open reduction and internal fixation in two hospitals, with a follow-up postal survey of 959 of the patients using three functional outcome scores. Obese patients had more severe overall complications and higher odds of any complication than the normal weight group, with adjusted odds ratio 1.67 (95% confidence interval (CI) 1.08 to 2.59; p = 0.021) and 1.71 (95% CI 1.10 to 2.65; p = 0.016), respectively. In total 479 patients (54.6%) responded to the questionnaire. Obese patients had worse scores on the Olerud and Molander Ankle Score (p < 0.001), Self-Reported Foot and Ankle Questionnaire (p = 0.003) and Lower Extremity Functional Scale (p = 0.01) than those with normal weight. In contrast, overweight patients did not have worse functional scores than those with normal weight. Obese patients had more complications, more severe complications, and worse functional outcomes three to six years after ankle surgery compared with those with normal weight. Cite this article: Bone Joint J 2017;99-B:1389-98.


Structure of the Ca(2+)-free Gla domain sheds light on membrane binding of blood coagulation proteins.
Reversible membrane binding of gamma-carboxyglutamic acid (Gla)-containing coagulation factors requires Ca(2+)-binding to 10-12 Gla residues. Here we describe the solution structure of the Ca(2+)-free Gla-EGF domain pair of factor x which reveals a striking difference between the Ca(2+)-free and Ca(2+)-loaded forms. In the Ca(2+)-free form Gla residues are exposed to solvent and Phe 4, Leu 5 and Val 8 form a hydrophobic cluster in the interior of the domain. In the Ca(2+)-loaded form Gla residues ligate Ca2+ in the core of the domain pushing the side-chains of the three hydrophobic residues into the solvent. We propose that the Ca(2+)-induced exposure of hydrophobic side chains is crucial for membrane binding of Gla-containing coagulation proteins.


Evidence for a susceptibility locus for panic disorder near the catechol-O-methyltransferase gene on chromosome 22.
A well-characterized single nucleotide polymorphism (472G/A-Val/Met-SNP8) in the coding sequence of the catechol-O-methyltransferase (COMT) gene leads to a three- to fourfold difference in enzymatic activity and clinical and animal studies suggest a role in anxiety states like panic disorder. Subjects from 70 panic disorder pedigrees, and 83 "triads", were genotyped at seven single nucleotide polymorphisms (SNPs), polymorphic microsatellites in the first intron of COMT and approximately 339kb upstream of COMT (D22S944) and analyzed for genetic association and linkage. Linkage analysis showed elevated LOD scores for 472G/A (SNP 8), silent exon 3 substitution (186C/T-SNP 5), and the marker D22S944 (2.88, 2.62, and 2.93, respectively), using a variety of diagnostic and genetic models. Association tests were not significant for the SNPs, but were highly significant for D22S944 (p =.0001-.0003). One three-marker haplotype formed from the above three polymorphisms was significantly associated with panic disorder (p =.0001), as was the "global" p value for this combination (p =.005). In addition, numerous haplotypes with combinations of D22S944 and COMT SNPs were found to be significantly associated with panic disorder. Our findings provide strong evidence for a susceptibility locus for panic disorder either within the COMT gene or in a nearby region of chromosome 22.


Targeted deletion of Hsf1, 2, and 4 genes in mice.
Heat-shock transcription factors (Hsfs) regulate transcription of heat-shock proteins as well as other genes whose promoters contain heat-shock elements. There are at least five Hsfs in mammalian cells, Hsf1, Hsf2, Hsf3, Hsf4, and Hsfy. To understand the physiological roles of Hsf1, Hsf2, and Hsf4 in vivo, we generated knockout mouse lines for these factors. In this chapter, we describe the design of the targeting vectors, the plasmids used, and the successful generation of mice lacking the individual genes. We also briefly describe what we have learned about the physiological functions of these genes in vivo.


Pitfalls of the peer world: how children cope with common rejection experiences.
This study examines the reliability and validity of a newly developed self-report measure designed to assess children's coping strategies in response to everyday rejection experiences. The Survey for Coping with Rejection Experiences (SCORE) was administered to 225 children and factor analysis of responses resulted in the conceptually meaningful subscales of active, aggressive, denial, and ruminative coping. A subset of 95 children also completed a battery of self- (depression, social anxiety), parent- (internalizing and externalizing problems), and peer- (social preference, aggression, withdrawal) ratings of socioemotional adjustment. Children's coping strategies were meaningfully associated with adjustment measures, although these associations differed for boys and girls in several instances. Importantly, both aggressive and ruminative coping strategies were positively associated with internalizing problems even after controlling for the effect of children's actual sociometric status. In addition, denial coping emerged as a significant moderator of the relation between status and adjustment.


Metabolic Profile of Growing Immune- and Surgically Castrated Iberian Pigs Fed Diets of Different Amino Acid Concentration.
The purpose of the current study was to further characterize the performance and nitrogen retention differences previously observed between immunocastrated (IC) and surgically castrated (SC) pure Iberian pigs. Fifty-four pigs were used (three sexes: IC males, IC females and SC males), fed three isoenergetic diets (160, 140 and 120 g CP/kg DM; six pigs/treatment combination) from 40 kg BW until slaughter (105 kg BW). Plasmatic post-absorptive metabolites and hormones, and backfat tissue composition were determined. The IC males showed a trend towards higher plasmatic creatinine (p = 0.06) and IGF-1 concentrations than SC males and IC females (p < 0.001). SC males showed a higher predisposition to carcass fatness and insulin resistance compared to IC males. Plasmatic isoleucine concentration was higher in IC and SC males compared to IC females (p < 0.01), and valine was greater in IC males compared to the rest of the pigs (p < 0.001). Total branched-chain amino acids (AA) were greater in IC males than in IC females (p < 0.01). Total plasmatic essential AA concentrations tended to be greater in IC males (p = 0.09). The present results support the increased performance and nitrogen retention capacity previously observed in IC male Iberian pigs compared to SC males and IC females, which can be attributed to increased anabolic capacity related with lean growth in Iberian IC males.


Preparation of lymphocytes from small volumes of peripheral mouse blood.
A method was developed which allowed for the isolation of lymphocytes (95% of leukocytes in the final suspensions) from small volumes of peripheral mouse blood (38% recovery). The method proved of value for studies of murine lymphocytes which required biochemical analysis of samples from individual animals on a serial basis.


An autoregulatory switch in sex-specific phf7 transcription causes loss of sexual identity and tumors in the Drosophila female germline.
Maintenance of germ cell sexual identity is essential for reproduction. Entry into the spermatogenesis or oogenesis pathway requires that the appropriate gene network is activated and the antagonist network is silenced. For example, in Drosophila female germ cells, forced expression of the testis-specific PHD finger protein 7 (PHF7) disrupts oogenesis, leading to either an agametic or germ cell tumor phenotype. Here, we show that PHF7-expressing ovarian germ cells inappropriately express hundreds of genes, many of which are male germline genes. We find that the majority of genes under PHF7 control in female germ cells are not under PHF7 control in male germ cells, suggesting that PHF7 is acting in a tissue-specific manner. Remarkably, transcriptional reprogramming includes a positive autoregulatory feedback mechanism in which ectopic PHF7 overcomes its own transcriptional repression through promoter switching. Furthermore, we find that tumorigenic capacity is dependent on the dosage of phf7 This study reveals that ectopic PHF7 in female germ cells leads to a loss of sexual identity and the promotion of a regulatory circuit that is beneficial for tumor initiation and progression.


A comparison of physician examiners and trained assessors in a high-stakes OSCE setting.
The Medical Council of Canada (MCC) administers an objective structured clinical examination for licensure. Traditionally, physician examiners (PE) have evaluated these examinees. Recruitment of physicians is becoming more difficult. Determining if alternate scorers can be used is of increasing importance. In 2003, the MCC ran a study using trained assessors (TA) simultaneously with PEs. Four examination centers and three history-taking stations were selected. Health care workers were recruited as the TAs. A 3x2x4 mixed analyses of variance indicated no significant difference between scorers (F1,462=.01, p=.94). There were significant interaction effects, which were, localized to site 1/station 3, site 3/station 2, and site 4/station1. Pass/fail decisions would have misclassified 14.4-25.01% of examinees. Trained assessors may be a valid alternative to PE for completing checklists in history-taking stations, but their role in completing global ratings is not supported by this study.


Phylogenetic analysis of encapsulated and non-encapsulated Trichinella species by studying the 5S rDNA tandemly repeated intergenic region.
The identification of sequence regions in the genomes of pathogens which can be useful to distinguish among species and genotypes, is of great importance for epidemiological, molecular, and phylogenetic studies. The 5S ribosomal DNA intergenic spacer region has been identified as a good target to distinguish among eight Trichinella species and genotypes. The recent discovery of two non-encapsulated species in this genus, Trichinella papuae and Trichinella zimbabwensis, which can infect both mammals and reptiles, has suggested analyzing their 5S rDNA. Amplification of the tandem repeats of the 5S rDNA intergenic region of encapsulated species of Trichinella shows a 751bp fragment, whereas the three non-encapsulated species show a fragment of 800bp with T. pseudospiralis showing an additional fragment of 522bp. Although the size of the 800bp PCR fragments of T. papuae and T. zimbabwensis are similar to that of T. pseudospiralis, there are differences in the 5S rDNA intergenic regions among the three non-encapsulated species. Phylogenetic analysis of the 5S rDNA intergenic regions shows a clustering together of the three non-encapsulated Trichinella species that is well separated from the encapsulated ones. In addition, a single PCR-based method allows distinguishing non-encapsulated and encapsulated species.


Effects of angiotensin II blockade on inflammation-induced alterations of pharmacokinetics and pharmacodynamics of calcium channel blockers.
Inflammation elevates plasma verapamil concentrations but diminishes pharmacological response. Angiotensin II is a pro-inflammatory mediator. We examined the effect of angiotensin II receptor blockade on the pharmacokinetics and pharmacodynamics of verapamil, as well as the binding properties and amounts of its target protein in calcium channels, in a rat model of inflammation. We used 4 groups of male Sprague-Dawley rats (220-280 g): inflamed-placebo, inflamed-treated, control-placebo and control-treated. Inflammation as pre-adjuvant arthritis was induced by injecting Mycobacterium butyricum on day 0. From day 6 to 12, 30 mg kg(-1) oral valsartan or placebo was administered twice daily. On day 12, a single oral dose of 25 mg kg(-1) verapamil was administered and prolongation of the PR interval measured and plasma samples collected for verapamil and nor-verapamil analysis. The amounts of the target protein Ca(v)1.2 subunit of L-type calcium channels in heart was measured by Western blotting and ligand binding with (3)H-nitrendipine. Inflammation reduced effects of verapamil, although plasma drug concentrations were increased. This was associated with a reduction in ligand binding capacity and amount of the calcium channel target protein in heart extracts. Valsartan significantly reversed the down-regulating effect of inflammation on verapamil's effects on the PR interval, and the lower level of protein binding and the decreased target protein. Reduced responses to calcium channel blockers in inflammatory conditions appeared to be due to a reduced amount of target protein that was reversed by the angiotensin II antagonist, valsartan.


Specific solute effects on Staphylococcus aureus cells subjected to reduced water activity.
The behaviour of Staphylococcus aureus cells subjected to lowered water activity (aw) by the addition of various solutes was examined by transmission electron microscopy. Solutes included sodium chloride, sucrose, propylene glycol, butylene glycol and various polyethylene glycols. Changes in other physical properties of the liquid growth medium brought about by the solutes were estimated theoretically. They did not correlate with the bacterial biological response. The inhibitory effects of sucrose and sodium chloride against S. aureus were primarily ascribed to their water activity-lowering abilities, showing no significant specific solute effects. However, the other solutes examined showed specific antibacterial activity against S. aureus which may be compatible with cell wall attack.


Long-term outcomes of arteriovenous snuffbox fistulas for hemodialysis.
Autologous arteriovenous fistulas in the arm have been shown to be superior to all other types of arteriovenous access and should be placed as far distally as possible. The aim of the present study was to delineate the advantages of snuffbox fistulas (SBFs). From January 2007 through April 2017, first time or new arteriovenous fistula surgery was performed in 375 patients. If feasible, an SBF was chosen. The observation period ended in December 2019. The focus of the present study was on the patency rates, redo surgery, complications, and mortality rates. SBFs were created in 236 patients (92 women and 144 men; mean age, 67 years; 46% with diabetes). Within the first 6 weeks, 11% of the fistulas had failed. The primary patency rates were 75%, 69%, 64%, 61%, 60%, and 58% after 1, 2, 3, 5, 7, and 9 years, respectively. The secondary patency rates were 86%, 83%, 80%, and 77% after 1, 2, 3, and 5 years, respectively. Age, sex, and the presence of diabetes mellitus had no effect on the primary patency rates. Patients with a right-sided SBF had significantly worse primary patency. Of the 236 patients, 154 (65%) had died during the observation period. The incidence of surgical site infection within the first 30 days and access-related ischemia during the observation period was <1%. The results from the present study have shown that with suitable anatomic conditions, a SBF is a good choice for the creation of dialysis access. The patency rates were encouraging, and it offers the longest puncturable segment of the cephalic vein. Also, the rate of perioperative complications was low.


A comparison of the safety and efficacy of lomefloxacin and ciprofloxacin in the treatment of complicated or recurrent urinary tract infections.
The efficacy and safety of two fluoroquinolone antimicrobial agents, lomefloxacin and ciprofloxacin, were compared in a randomized, investigator-blinded, multiple-dose study that enrolled 150 adult outpatients with complicated or recurrent urinary tract infections. A total of 75 patients were treated with 400 mg of lomefloxacin once daily for 10-14 days; an equal number of patients received 500 mg of ciprofloxacin every 12 hours for 10-14 days. Both groups of patients were comparable in terms of demographics and distribution of underlying conditions. The most frequently occurring pathogens in both groups were Escherichia coli and Klebsiella pneumoniae. A total of 142 patients met the criteria for efficacy evaluation, 72 in the lomefloxacin group and 70 in the ciprofloxacin group. Eradication of the initial pathogen (post-treatment culture of less than or equal to 10(4) colony-forming units (CFU)/mL) was noted in 97.2% of patients treated with lomefloxacin and in 95.7% of ciprofloxacin-treated patients. Clinical success (disappearance or amelioration of presenting signs and symptoms) occurred in 98.6% of lomefloxacin-treated patients and in 95.7% of patients treated with ciprofloxacin. The differences in outcome between the two treatment groups were not statistically significant. Both drug regimens were well tolerated. There were no clinically significant changes in clinical laboratory values during treatment. In the lomefloxacin group three patients reported nausea, one nervousness, and one pruritus; in the ciprofloxacin group, two patients reported nausea and three pruritus. Two lomefloxacin-treated patients (3%) and four patients treated with ciprofloxacin (5%) withdrew from treatment because of adverse events. A single daily dose of 400 mg lomefloxacin was comparable to 500 mg ciprofloxacin every 12 hours for the treatment of complicated or recurrent urinary tract infections.


Expression pattern of Piwi-like genes in adult Myzostoma cirriferum (Annelida).
Piwi-like genes are a subgroup of Argonaute genes which participate as gene regulators by gene silencing. In most bilaterians, such as mouse, human, insects, and zebrafish, their expression is mostly limited to gonadal stem cells. But there are some striking exceptions to this pattern; flatworms and acoels also express Piwi-like genes in somatic stem cells, due to their unique replacement system. Annelid species like Capitella teleta and Platynereis dumerilii express these genes in cells of the posterior growth zone as well as in gonadal stem cells. To investigate the expression pattern of Piwi-like genes in another annelid, we established in situ hybridization for adult Myzostoma cirriferum. Piwi-like gene transcripts recovered in an mRNA-seq library of pooled adult stages of M. cirriferum were expanded using RACE PCR, cloned and sequenced. ML analysis confirmed the identity of both transcripts as part of the Piwi1-like or Piwi2-like subfamily of Argonaute proteins. The results of in situ hybridization studies show that the expression of both Piwi-like genes, Mc-Piwi1 and Mc-Piwi2, is clearly located only in gonadal stem cells, and as such we did not find any evidence for the existence of a posterior growth zone nor expression in somatic stem cells.


[Congenital radial dysplasia (thumb ray and radial carpus)--an atavism].
A case of hypoplastic thumb with anomalies of the radial carpal bones is reported, believed to be atavistic in origin.


Evaluation of microbiological accumulation capability of the commercial sponge Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae).
This study was carried out to evaluate the microbiological accumulation capability of the demosponge Spongia officinalis var. adriatica. Six microbiological parameters were researched in two sampling periods in the water and in reared sponge samples coming from sites with different degrees of microbial contamination: an off-shore fish farm displaced off the Apulian coast (Southern Adriatic Sea) and a no-impacted area displaced into the Marine Protected Area of Porto Cesareo (Apulian coast-Ionian Sea). We detected the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 degrees C on Plate Count Agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total and fecal coliforms as well as fecal streptococci concentrations were detected by the MPN method. Bacterial densities were always higher in the sponge homogenates compared with the corresponding seawater in the sampling points and in both sampling periods. As regard vibrios, total culturable bacteria at 37 degrees C and fecal streptococci concentrations, the highest values were observed in the sponge samples coming from the off-shore fish farm during the summer period. The ability of Spongia officinalis var. adriatica to accumulate the microbial pollution indicators suggests that this species can be employed as a bioindicator for monitoring water quality.


Dr. Johnstone et al. Reply to Dr. Hamilton.
We thank Dr. Hamilton1 for his interest in our research and for provoking a more nuanced and detailed approach to analyzing the relationship among treatment assignment, treatment response, and correct treatment guessing in randomized controlled trials; in this case, the Micronutrients for ADHD in Youth (MADDY) study.2.


Studies show link between meth use and HIV infections. Resurgence of STDs tied to drug's use.
Meth use is associated with HIV infection and high rates of sexually transmitted diseases among men who have sex with men in Los Angeles and New York City. But it's also associated with high-risk sexual and infection behavior among rural white women and other heterosexuals in Tennessee, new research shows. Researchers discuss the problems they've encountered with meth use and how it has been linked to HIV infection in some populations.


Predictors of acute and long-term success after radiofrequency catheter ablation of type 1 atrial flutter.
A history of atrial fibrillation and right atrial size are both determinants of procedural and long-term outcome after RF catheter ablation of type 1 atrial flutter. The results of this study suggest that patients with no history of atrial fibrillation and a normal right atrial size have the best short- and long-term results from this procedure.


Combining allele-specific fluorescent probes and restriction assay in real-time PCR to achieve SNP scoring beyond allele ratios of 1:1000.
TaqMan-nuclease assays are widely used for the qualitative detection of single nucleotide polymorphisms (SNPs) and the determination of biallelic states in pooled or heterozygous DNA samples. These assays are highly specific, reproducible, and suitable for high-throughput approaches. A crucial limitation of this method, and others, is the detection qf minor allele frequencies with detection limits of generally 3% to 9% for minor allele contributions. Here we describe the combination of customized TaqMan-nuclease assay and allele-specific restriction to increase the sensitivity of this method, allowing the qualitative detection of allele contributions as low as 0.05%.


Methods and Patterns for User-Friendly Quantum Programming.
The power and efficiency of particular quantum algorithms over classical ones has been proved. The rise of quantum computing and algorithms has highlighted the need for appropriate programming means and tools. Here, we present a brief overview of some techniques and a proposed methodology in writing quantum programs and designing languages. Our approach offers "user-friendly" features to ease the development of such programs. We also give indicative snippets in an untyped fragment of the Qumin language, describing well-known quantum algorithms.


Photocatalytic CO2 -to-Syngas Evolution with Molecular Catalyst Metal-Organic Framework Nanozymes.
Syngas, a mixture of CO and H2 , is a high-priority intermediate for producing several commodity chemicals, e.g., ammonia, methanol, and synthetic hydrocarbon fuels. Accordingly, parallel sunlight-driven catalytic conversion of CO2 and protons to syngas is a key step toward a sustainable energy cycle. State-of-the-art catalytic systems and materials often fall short as application-oriented concurrent CO and H2 evolution requires challenging reaction conditions which can hamper stability, selectivity, and efficiency. Here a light-harvesting metal-organic framework hosting two molecular catalysts is engineered to yield colloidal, water-stable, versatile nanoreactors for photocatalytic syngas generation with highly controllable product ratios. In-depth fluorescence, X-ray, and microscopic studies paired with kinetic analysis show that the host delivers energy efficiently to active sites, conceptually yielding nanozymes. This unlocked sustained CO2 reduction and H2 evolution with benchmark turnover numbers and record incident photon conversions up to 36%, showcasing a highly active and durable all-in-one material toward application in solar energy-driven syngas generation.


Comparison of HER2 Expression in Primary Tumor and Disseminated Tumor Cells in the Bone Marrow of Breast Cancer Patients.
The aim of this study was to measure the human epidermal growth factor receptor 2 (HER2) status of disseminated tumor cells (DTCs) from bone marrow (BM) aspirates and to assess correspondence or discrepancy with the primary tumor. DTCs were isolated from the BM of 156 breast cancer patients. Cytokeratin-positive DTCs were further analyzed by the chromogenic in situ hybridization method to detect HER2 gene amplification. A significant correlation (p = 0.021) was found between the HER2 status of DTCs and the primary tumors. Sixty-one (68.5%) patients had a corresponding status. However, a shift of phenotype between primary tumor and DTCs was found in the remaining patients. This study showed a significant grade of discordance of the HER2 status between primary tumors and DTCs in the BM of a relevant subgroup of patients. Detection of HER2 amplification on DTCs could therefore help to better stratify patients for a more tailored therapy, since they would benefit from a HER2-targeted therapy.


Substrate-typic changes of platelet monoamine oxidase activity in sub-types of schizophrenia.
Monoamine oxidase (MAO) activity has been measured in the platelets of controls (n = 42) and schizophrenic patients (n = 49) of three subtypes, using beta-phenylethylamine, p-tyramine, and tryptamine as substrates. Characteristic differences of MAO activity were observed between platelets of patients and controls; the differences were substrate-typic: decreased enzyme activity was found with all three substrates in platelets of the parnaoid subtype. With tryptamine, MAO activity was decreased in the platelets of all three sub-types of schizophrenia. With p-tyramine, MAO was low in patients with affective psychoses and paranoid schizophrenia. The value of MAO activity measurements as a means for distinguishing sub-types of schizophrenic disorders is improved by using two substrates; tryptamine and p-tyramine. Possible mechanisms of the substrate-typic changes of platelet MAO activity in schizophrenia are discussed.


Synergism of type I and type II interferons in stimulating the activity of the same DNA enhancer.
Type I and type II interferons (IFNs) can act synergistically to activate the transcription of the 2-5A synthetase gene. We used in vivo functional assays of sequences from the gene promoter region to determine which DNA segment mediates the gene induction by IFN gamma and the synergistic effect. We found that the type I IFN-inducible enhancer (or IRS) of the 2-5A synthetase gene also confers inducibility by type II IFN to a reporter CAT gene, though the time course and dose response of the induction by the two IFNs are quite different. A clear synergism of the two IFN in stimulating the IRS is observed at low doses of the two IFNs.


Enhancement of antibacterial superoxide-anion generation in human monocytes by fumaric acid esters.
Fumaric acid esters (FAE) are used for the systemic therapy of psoriasis with high clinical efficacy. Among the potential side effects of FAE therapy, lymphocytopenia is sometimes observed. We have investigated the effect of dimethylfumarate (DMF) and its main metabolite methylhydrogenfumarate (MHF) as well as dexamethasone on superoxide anion generation by human monocytes and neutrophils after stimulation with bacteria (Staphylococcus aureus and Escherichia coli) and the yeast Candida albicans in addition with zymosan particles and with the tripeptide fMLP. Expression of mannose receptors on monocytes and neutrophils was also analyzed. The results showed that dexamethasone significantly inhibited superoxide anion generation from monocytes in response to bacteria and C. albicans, whereas DMF as well as MHF dose dependently increased the production of superoxide anion in monocytes in response to zymosan, fMLP and bacteria. Dexamethasone, DMF or MHF did not modulate superoxide anion generation of neutrophils. Expression of mannose receptors on monocytes was not regulated by DMF or MHF. Our data provide evidence that DMF and MHF do not alter the production of superoxide anions as an important mechanism of innate defense against microorganisms.


Anaphylactic shock due to cefuroxime in a patient taking penicillin prophylaxis.
We report a case of cardiovascular collapse in a homozygous sickle cell patient undergoing elective orthopaedic surgery, following the intravenous administration of cefuroxime. This patient had no known previous drug allergies. She had been taking prophylactic penicillin V on a daily basis, for 10 years and had encountered no problems. She had received cefuroxime on at least four previous occasions, for the treatment of chest infections, with no reported problems. Her serum mast cell tryptase, 1 h after the onset of anaphylaxis, was greater than 200 microg x l(-1) (normal range 10-16). She was skin prick tested, 6 weeks after the event, to all the drugs administered during the course of the anaesthetic and reacted positively only to cefuroxime. Specific IgE levels for penicillin determinants and latex were negative (13 and 0 IU x ml(-1), respectively UniCAP Pharmacia & Upjohn).


Evaluation of different intraarticular injection volumes to assess optimum efficient amount; an experimental study in rat knee joints.
The injection amount of solutions differs in wide ranges, from 20 to 200 μL in intraarticular injections experiments that was carried out with rats. The 10-fold difference between applied volumes is not acceptable since injection errors might affect the outcomes of the studies significantly. The excessive amount of solution that was injected into the knee joint of rats might end up with leakage of the solution from injection aperture, and damage the articular structures etc. In our preliminary experiments, we mentioned problems during the injection of excessive amount of solution into the rat knee joint. The main purpose of the present study, which was performed with rat knee joints, was to evaluate the applicability of different amounts of solutions used in experiments and to determine the optimal volume for intraarticular injection in rats. The volume of 100 μL water-based stain was administered into knee joints of non-living rats. The legs were dissected to evaluate the possible presence of injected solution outside the knee joint. Then, the decreasing volumes (100 μL, 50 μL, 40 μL and 30 μL) of X-ray contrast solution (lobitridol) were applied into the knees of living rats to investigate whether systemic circulation acts on the leakage outside the joint. The X-ray images of all these joints were also taken to confirm the results. In nonliving rats, the place where water-based strain overflow from the knee joint was pointed out. The place of leakage is where the tendo musculi extensor digitorum longus leaves outside the joint. In living rats, by using contrast solution, an overflow was noticed in the subdermal place with the intraarticular injection volumes of 100 μL, 50 μL, and 40 μL. No overflow was observed with the intraarticular injection volumes of 30 μL X-ray contrast solution. In conclusion, the injection volume for the knee joints of rats should not exceed the 30 μL for optimum efficiency.


The labile lipid fraction of meat: from perceived disease and waste to health and opportunity.
The fatty acid composition of beef and pork has been stigmatized due to their relationships with several diseases from cardiovascular disease to cancer. Meat lipids are, however, one of the few components of meat that can be modified in content and composition, and can present opportunities for value added production and health promotion. Until regulations and policies are in place to define requirements for fatty acid enrichment, however, the process remains relatively academic. Once practical goals are in place for fatty acid enrichment in meat, both theory and practice need to converge for successful production of fatty acid enriched meat. The present review covers aspects of policy in Canada, and requirements for research networks to respond to theoretical and practical challenges associated with production of fatty acid enriched meat. Finally, needs for education and marketing are outlined which must be in place to truly realize a transition of meat lipids from perceived disease and waste to health and opportunity.


Dynamic expression pattern of Nodal-related genes during left-right development in medaka.
Nodal-related genes have been implicated in mesendoderm induction, establishment of embryonic axes, neural patterning and left-right development among vertebrates. Here we report the isolation of three Nodal-related genes in medaka (Oryzias latipes). Based on sequence analysis and in accordance to zebrafish orthologues we named the isolated genes Ndr1, Ndr2 and Spaw. Gene expression analysis throughout medaka development confirmed this assignment. Ndr1 and Ndr2 are detectable during gastrulation whereas Ndr2 and Spaw are expressed asymmetrically during somitogenesis. In accordance with its zebrafish orthologue, Spaw is expressed as the first asymmetric marker in the left lateral plate mesoderm (LPM) and Ndr2 displays asymmetric expression domains in the brain and the LPM. In general, the spatial distribution of Nodal transcripts resembles those reported for zebrafish, in case of Ndr2, however, we report a novel left-right asymmetry in the posterior paraxial mesoderm flanking the Kupffer's vesicle.


[Muscle weakness].
A 61 year old man complained initially about pain in the left lower leg with increasing swelling of the entire left leg. A slight swelling of the left ankle and knee and an erythematous skin change of 5 cm diameter developed and disappeared after a few hours. Two days later the patient complained about a sudden and massive weakness of the proximal skeletal muscles, progressive disturbance of swallowing, hoarseness and dark red to bluish erythematous skin eruptions on the right knee and buttock. An elevated creatinine-kinase and lactate-dehydrogenase were found. Dermatomyositis was diagnosed and treatment with high dose steroids initiated. Under this treatment the patients condition deteriorated dramatically. A treatment with plasmapheresis and cyclophosphamide was begun. A significant improvement occurred over three weeks. The patient was discharged, rehabilitated and remains free of symptoms on 10 mg cortisone. A neoplasm has so far not been detected.


Neutralizing Monoclonal Antibodies against Disparate Epitopes on Ricin Toxin's Enzymatic Subunit Interfere with Intracellular Toxin Transport.
Ricin is a member of the A-B family of bacterial and plant toxins that exploit retrograde trafficking to the Golgi apparatus and endoplasmic reticulum (ER) as a means to deliver their cytotoxic enzymatic subunits into the cytoplasm of mammalian cells. In this study we demonstrate that R70 and SyH7, two well-characterized monoclonal antibodies (mAbs) directed against distinct epitopes on the surface of ricin's enzymatic subunit (RTA), interfere with toxin transport from the plasma membrane to the trans Golgi network. Toxin-mAb complexes formed on the cell surface delayed ricin's egress from EEA-1(+) and Rab7(+) vesicles and enhanced toxin accumulation in LAMP-1(+) vesicles, suggesting the complexes were destined for degradation in lysosomes. Three other RTA-specific neutralizing mAbs against different epitopes were similar to R70 and SyH7 in terms of their effects on ricin retrograde transport. We conclude that interference with toxin retrograde transport may be a hallmark of toxin-neutralizing antibodies directed against disparate epitopes on RTA.


Development of a duplex rapid assay for immunoglobulins M and G to evaluate the parvoviral immune status of clinically healthy dogs.
A duplex rapid assay for detection of serum antibodies to canine parvovirus (CPV) was developed. Canine immunoglobulin (Ig)M or IgG were captured in immunotubes with anti-canine IgM or IgG and detected with parvovirus VP2 recombinant protein followed by an anti-VP2 monoclonal antibody. The assay was tested using a collection of sera from dogs that were vaccinated against CPV on arrival at an animal shelter in Madrid, Spain. Results were compared with those of 2 commercial enzyme-linked immunosorbent assays (ELISAs) considered as reference techniques. A high correlation was found between the duplex rapid assay and the ELISAs, presenting an accuracy of 98% and 100% for IgG and IgM, respectively. According to the IgG and IgM levels at days 0-3 postvaccination, the samples were divided into 2 groups. One group of dogs showed high IgG and low IgM values at the first sampling post-vaccination and during the following 14 days, indicating that they had previously been in contact with the virus, either by vaccination or infection before arrival at the animal shelter. A second group of dogs appeared to be unvaccinated or uninfected before arrival at the animal shelter because they had negative IgM and IgG values soon after vaccination. These animals responded to vaccination, as demonstrated by seroconversion of both isotypes of immunoglobulins. The developed assay appears to be useful in determining the unknown immune status of dogs to CPV, especially in kennels and shelters where the rate of infection by CPV is relatively high.


Interfacial construction of SnS2/Zn0.2Cd0.8S nanopolyhedron heterojunctions for enhanced photocatalytic hydrogen evolution.
ZnCdS, a metal chalcogenide solid solution, has attracted significant attention. However, two primary challenges hinder its widespread application in photocatalytic hydrogen evolution: the rapid recombination rate of photogenerated carriers and susceptibility to photo-oxidation in the aqueous environments. In this article, a facile hydrothermal route was employed for the first time to uniformly assemble SnS2 nanoparticles onto the surface of Zn0.2Cd0.8S (ZCS) nanopolyhedra. The intimate contact of two materials resulted in the formation of heterojunctions. By adjusting the content of SnS2, the hydrogen evolution reaction (HER) performance was optimized to reach up to 12170 μmol/gh, which is 1.9 times of the pristine ZCS. Notably, the photocatalyst demonstrated remarkable stability with an apparent quantum yield (AQY) of 15.5% at 420 nm. The enhanced photocatalytic performance can be attributed to the following factors: (i) The heterojunction composite, with larger surface area and more micropores, provides additional active sites and exhibits high photocatalytic activity; (ii) The internal electric field accelerates the separation of photogenerated carriers and reduces the recombination rate of electron-hole pairs; (iii) The photogenerated holes can be quickly transferred to the valence band of SnS2 and react with triethanolamine, thereby significantly reducing the photo-oxidation of ZCS. This work not only proposed a feasible route to improve the photocatalytic activity of ZCS, but also provided insights into the role of heterojunctions and the reaction mechanism.


Developing data-driven clinical pathways using electronic health records: The cases of total laparoscopic hysterectomy and rotator cuff tears.
A clinical pathway is one of the tools used to support clinical decision making that provides a standardized care process in a specific context. The objective of this research was to develop a method for building data-driven clinical pathways using electronic health record data. We proposed a matching rate-based clinical pathway mining algorithm that produces the optimal set of clinical orders for each clinical stage by employing matching rates. To validate the approach, we utilized two different datasets of deidentified inpatient records directly related to total laparoscopic hysterectomy (TLH) and rotator cuff tears (RCTs) from a hospital in South Korea. The derived data-driven clinical pathways were evaluated with knowledge-based models by health professionals using a delta analysis. Two different data-driven clinical pathways, i.e., TLH and RCTs, were produced by applying the matching rate-based clinical pathway mining algorithm. We identified that there were significant differences in clinical orders between the data-driven and knowledge-based models. Additionally, the data-driven clinical pathways based on our algorithm outperformed the models by clinical experts, with average matching rates of 82.02% and 79.66%, respectively. The proposed algorithm will be helpful for supporting clinical decisions and directly applicable in medical practices.


Interactive effects of age and recent substance use on striatal shape morphology at substance use disorder treatment entry.
Striatal neuroadaptations are regarded to play an important role in the progression from voluntary to compulsive use of addictive substances and provide a promising target for the identification of neuroimaging biomarkers. Recent advances in surface-based computational analysis enable morphological assessment linking variations in global and local striatal shape to duration and magnitude of substance use with a degree of sensitivity that exceeds standard volumetric analysis. This study used a new segmentation methodology coupled with local surface-based indices of surface area and displacement to provide a comprehensive structural characterization of the striatum in 34 patients entering treatment for substance use disorder (SUD) and 49 controls, and to examine the influence of recent substance use on abnormal age-related striatal deformation in SUD patients. Patients showed a small reduction in striatal volume and no difference in surface area or shape in comparison to controls. Between-group differences in shape were likely neutralized by the bidirectional influence of recent substance use on striatal shape in SUD patients. Specifically, there was an interaction between age and substance such that among older patients more drug use was associated with greater inward striatal contraction but more alcohol use was associated with greater outward expansion. This study builds on previous work and advances our understanding of the nature of striatal neuroadaptations as a potential biomarker of disease progression in addiction.


[Retrocaval ureter in children: surgical approach based on the obstructive pattern in the diuretic renogram with 99mTc DTPA].
Retrocaval ureter is a rare disease in childhood which is due to a abnormal development of the inferior cava vein. We report a new case of retrocaval ureter in a 9 year-old girl who had consulted for macrohematuria and right flank pain. Preoperative intravenous urography and DTPA diuretic renogram suggested this entity. CT scan was no necessary for the diagnosis. We consider the diuretic renogram the best diagnostic method in the surgical decision of this disease.


Longitudinal study of inpatients admitted to a general activation service: variables that predict discharge to a patient's discharge goal location.
To determine which sociodemographic and clinical characteristics of patients admitted to a general activation service (GAS) are predictive of discharge to patients' discharge goal locations (DGLs). Prospective cohort study. Rehabilitation and complex continuing care hospital in southern Ontario, Canada. Patients admitted from January 2000 to December 2002 (N=154). The GAS. Patients indicated on their service applications where they wanted to be discharged. This is termed the DGL. Fifty-three percent of the sample were discharged to their DGLs. Ninety-eight percent of these patients were discharged by 9 months. Eighty-seven percent who were discharged to their DGLs were discharged to their own home. Predictors of being discharged to the DGL were better activities of daily living scores, good vision, and having sufficient help at home. Expert clinician opinion of the likelihood of each patient being discharged to his/her DGL, based on initial assessment, was also predictive of each patient's eventual discharge to his/her DGL. The GAS has a 53% success rate in discharging patients to their DGLs. Variables have been identified that should be useful in predicting whether patients will be discharged to their DGLs. Our findings are meaningful and informative in determining future admission criteria for the service.


The Renal Pathology of Obesity: Structure-Function Correlations.
The kidney is one of the target organs that may show health disorders as a result of obesity. Obesity-related glomerulopathy (ORG) is a kidney disease category based on a biopsy diagnosis that may occur secondary to obesity. Detailed clinicopathologic observations of ORG have provided significant knowledge regarding obesity-associated renal complications. Glomerulomegaly with focal segmental glomerulosclerosis of perihilar locations is a typical renal histopathologic finding in ORG, which has long been considered to represent a state of single-nephron glomerular hyperfiltration. This hypothesis was recently confirmed in ORG patients by estimating single-nephron glomerular filtration rate using a combined image analysis and biopsy-based stereology. Overshooting in glomerulotubular and tubuloglomerular interactions may lead to glomerular hyperfiltration/hypertension, podocyte failure, tubular protein-traffic overload, and tubulointerstitial scarring, constituting a vicious cycle of a common pathway to the further loss of functioning nephrons and the progression of kidney functional impairment.


Recent developments in the understanding and use of anthrax vaccine adsorbed: achieving more with less.
Anthrax Vaccine Adsorbed (AVA, BioThrax™) is the only Food and Drug Administration (FDA) approved vaccine for the prevention of anthrax in humans. Recent improvements in pre-exposure prophylaxis (PrEP) use of AVA include intramuscular (IM) administration and simplification of the priming series to three doses over 6 months. Administration IM markedly reduced the frequency, severity and duration of injection site reactions. Refinement of animal models for inhalation anthrax, identification of immune correlates of protection and cross-species modeling have created opportunities for reductions in the PrEP booster schedule and were pivotal in FDA approval of a post-exposure prophylaxis (PEP) indication. Clinical and nonclinical studies of accelerated PEP schedules and divided doses may provide prospects for shortening the PEP antimicrobial treatment period. These data may assist in determining feasibility of expanded coverage in a large-scale emergency when vaccine demand may exceed availability. Enhancements to the AVA formulation may broaden the vaccine's PEP application.


Primary Care Behavioral Health (PCBH) Model Research: Current State of the Science and a Call to Action.
The Primary Care Behavioral Health (PCBH) model of service delivery is being used increasingly as an effective way to integrate behavioral health services into primary care. Despite its growing popularity, scientifically robust research on the model is lacking. In this article, we provide a qualitative review of published PCBH model research on patient and implementation outcomes. We review common barriers and potential solutions for improving the quantity and quality of PCBH model research, the vital data that need to be collected over the next 10 years, and how to collect those data.


The study on photoreflectance characteristic of semi-insulating GaAs surface region by its exposure to 6 MeV electron beam.
Photoreflectance measurements were performed to investigate the optical properties in the electron beam irradiation semi-insulating GaAs(e-beam irradiation GaAs) and semi-insulating GaAs(SI-GaAs). A considerable increase of the PR amplitudes has been registered after the e-beam irradiation in comparison with the GaAs. It is that result of a higher electron scattering on the lattice defects created by the e-beam.


Nitric oxide synthase expression correlates with death in an experimental mouse model of dengue with CNS involvement.
The clinical presentation of dengue is classified by the World Health Organization into dengue without warning signs, dengue with warning signs and severe dengue. Reports of neurological disease caused by Dengue virus (DENV) are becoming frequent, with symptoms that include reduced consciousness, severe headache, neck stiffness, focal neurological signs, tense fontanelle and convulsions. However, the immune mechanisms involved in neurovirulence remain poorly understood. Here we present a mouse model in which one genotype of DENV is inoculated by the intracranial route and infects C57/BL6 mice and replicates in the brain, causing death of mice. Mice were infected with different serotypes/genotypes of DENV by the intracranial route to evaluate viral replication, host cytokine and nitric oxide synthase 2 (Nos2) expression in the brain via real-time PCR. Histological analysis of the brain tissues was also performed. An analysis of which cells were responsible for the expression of cytokines and Nos2 was performed using flow cytometry. Survival curves of infected animals were also generated DENV 3 genotype I infected mice and replicated in the brain, causing death in our murine model. The increased levels of NOS2 could be the cause of the death of infected mice, as viral replication correlates with increased Nos2 and cytokine expression in the brain of C57BL/6 mice. In Nos2-/- mice that were infected with DENV, no clinical signs of infection were observed and cytokines were expressed at low levels, with the exception of interferon gamma (Ifng). Additionally, the Ifng-/- mice infected with DENV exhibited a severe and lethal disease, similar to the disease observed in C57BL/6 mice, while the DENV- infected Nos2-/- mice did not display increased mortality. Analyses of the brains from infected C57BL/6 mice revealed neuronal degeneration and necrosis during histopathologic examination. IFNg and NOS2 were produced in the brains of infected mice by CD4+ T cells and macrophages, respectively. The neurovirulence of DENV 3 genotype I is associated with a deleterious role of NOS2 in the brain, confirming this murine model as an appropriate tool to study DENV neurovirulence.


Vascular permeability precedes spontaneous intracerebral hemorrhage in stroke-prone spontaneously hypertensive rats.
Stroke-prone spontaneous hypertensive rats (SHRsp) fed a high-salt diet develop malignant hypertension, blood-brain barrier breakdown, and spontaneous intracerebral hemorrhage (ICH). The precise spatial and temporal relationship between these events has not been well-delineated. Ten SHRsp male rats, fed a high-salt diet, were imaged weekly using MRI, starting at 12 weeks of age. T1-weighted (with and without Gd), T2-weighted, and T2* sequences were acquired. Permeability maps were calculated. Seven SHRsp rats had spontaneous ICH develop before death. Five of the 7 rats had focally increased vascular permeability at the site of the ICH; 3 of these rats had vascular permeability 1 to 2 weeks before spontaneous ICH. Salt-loaded SHRsp rats have increased vascular permeability up to 2 weeks before ICH, predicting hemorrhage both in space and time. These results suggest that hypertensive ICH is preceded by focal vasculopathy detectable by Gd leak.


An efficient one-pot synthesis generating 4-ene-3,6-dione functionalised steroids from steroidal 5-en-3beta-ols using a modified Jones oxidation methodology.
Steroids with 4-ene-3,6-dione functionality have application in natural product chemistry, as synthetic intermediates and as aromatase inhibitors. Here, we report a two-phase oxidation of a range of steroidal 5-en-3beta-ols into corresponding 4-ene-3,6-diones using a modified Jones oxidation. The new reaction affords high yields (77-89%) of product in relatively short reaction times (1-2h). The simplicity of this reaction gives significant advantages over previously reported methodologies.


Reversible paralysis and loss of deep pain sensation after topical intrathecal morphine administration following durotomy.
To report 2 dogs that developed temporary pelvic limb paralysis with loss of deep pain sensation after topical intrathecal morphine administration during spinal surgery. Clinical report. Dogs (n = 2). A 5-year-old castrated male French Bulldog with a subarachnoid diverticulum at T9-T10 and a 9.5-year-old castrated male Belgian Shepherd dog with a herniated disc at T12-T13 and intradural component, had Gelfoam soaked with morphine placed over the dura mater defect. Pelvic limb paralysis and loss of deep pain sensation was noticed immediately after recovery from anesthesia. After intravenous naloxone administration, both dogs immediately regained ambulation and normal pain sensation; however the effect was temporary lasting only a few hours. Permanent resolution of clinical signs occurred 24 hours after surgery. Topical intrathecal morphine administration resulted in temporary pelvic limb paralysis and loss of deep pain sensation. This route of administration should be used cautiously until further determination of the efficacy and adverse effects associated with topical intrathecal morphine administration.


Synthesis, structural analysis, and docking studies with SARS-CoV-2 of a trinuclear zinc complex with N-phenylanthranilic acid ligands.
The structure of a trinuclear zinc complex, hexakis(μ2-2-anilinobenzoato)diaquatrizinc(II), [Zn2(C13H10NO2)6(H2O)2] or (NPA)6Zn3(H2O)2 (NPA is 2-anilinobenzoate or N-phenylanthranilate), is reported. The complex crystallizes in the triclinic space group P-1 and the central ZnII atom is located on an inversion center. The NPA ligand is found to coordinate via the carboxylate O atoms with unique C-O bond lengths that support an unequal distribution of resonance over the carboxylate fragment. The axial H2O ligands form hydrogen bonds with neighboring molecules that stabilize the supramolecular system in rigid straight chains, with an angle of 180° along the c axis. π stacking is the primary stabilization along the a and b axes, resulting in a highly ordered supramolecular structure. Docking studies show that this unique supramolecular structure of a trinuclear zinc complex has potential for binding to the main protease (Mpro) in SARS-CoV-2 in a different location from Remdesivir, but with a similar binding strength.


High Resolution Structures of the Human ABO(H) Blood Group Enzymes in Complex with Donor Analogs Reveal That the Enzymes Utilize Multiple Donor Conformations to Bind Substrates in a Stepwise Manner.
Homologous glycosyltransferases α-(1→3)-N-acetylgalactosaminyltransferase (GTA) and α-(1→3)-galactosyltransferase (GTB) catalyze the final step in ABO(H) blood group A and B antigen synthesis through sugar transfer from activated donor to the H antigen acceptor. These enzymes have a GT-A fold type with characteristic mobile polypeptide loops that cover the active site upon substrate binding and, despite intense investigation, many aspects of substrate specificity and catalysis remain unclear. The structures of GTA, GTB, and their chimeras have been determined to between 1.55 and 1.39 Å resolution in complex with natural donors UDP-Gal, UDP-Glc and, in an attempt to overcome one of the common problems associated with three-dimensional studies, the non-hydrolyzable donor analog UDP-phosphono-galactose (UDP-C-Gal). Whereas the uracil moieties of the donors are observed to maintain a constant location, the sugar moieties lie in four distinct conformations, varying from extended to the "tucked under" conformation associated with catalysis, each stabilized by different hydrogen bonding partners with the enzyme. Further, several structures show clear evidence that the donor sugar is disordered over two of the observed conformations and so provide evidence for stepwise insertion into the active site. Although the natural donors can both assume the tucked under conformation in complex with enzyme, UDP-C-Gal cannot. Whereas UDP-C-Gal was designed to be "isosteric" with natural donor, the small differences in structure imposed by changing the epimeric oxygen atom to carbon appear to render the enzyme incapable of binding the analog in the active conformation and so preclude its use as a substrate mimic in GTA and GTB.


A Pilot Study Examining Factors Influencing Readiness to Progress to Indirect Supervision Among First Year Residents in a General Psychiatry Training Program.
In the first year of training, psychiatry residents progress from direct supervision to indirect supervision but factors predicting time to transition between these levels of supervision are unknown. This study aimed to examine times for transition to indirect levels of supervision and to identify resident factors associated with slower progression. The authors compiled data from training files from years 2011-2015, including licensing exam scores, age, gender, medical school, month of first inpatient psychiatry rotation, and transition times between levels of supervision. Correlational analysis examined the relationship between these factors. Univariate analysis further examined the relationship between medical school training and transition times between supervision levels. Among the factors studied, only international medical school training was positively correlated with time to transition to indirect supervision and between levels of indirect supervision. International medical graduate (IMG) interns in psychiatry training may benefit from additional training and support to reach competencies required for the transition to indirect supervision.


Complete Genome Sequence of Lagilactobacillus salivarius VHProbi A17, Isolated from Fecal Samples.
The Lagilactobacillus salivarius strain VHProbi A17 is a bacterium that is found to reside in the intestinal tract. Whole-genome-sequencing-based analysis is an ideal approach for evaluating its probiotic properties. Here, we report the whole-genome sequence of this strain, which contains a chromosome and four plasmids, with GC contents of 32.86%.


Preoperative radiation therapy in clinical stage II endometrial carcinoma.
From 1980 to 1987, 30 patients with FIGO clinical Stage II carcinoma of the endometrium were treated with 5000 cGy preoperative pelvic radiation therapy at Thomas Jefferson University Hospital. Patients with gross cervical disease received additional intracavitary irradiation with a tandem and ovoids for a combined total dose of 7000 cGy to point A. All patients then underwent exploratory laparotomy, total abdominal hysterectomy, and bilateral salpingo-oophorectomy (TAH/BSO). The 5-year actuarial survival for the entire group was 69%. The 5-year actuarial survival for the 8 patients with papillary serous, clear cell, and undifferentiated small cell carcinoma was 38%, with most patients failing in the upper abdomen. The 5-year actuarial survival for the remaining 22 patients was 82%. The only local failure occurred in the patient with an undifferentiated small cell carcinoma. Although preoperative pelvic radiation therapy together with TAH/BSO appears to offer excellent local control in all patients with Stage II endometrial carcinoma, additional treatment options should be considered for patients with papillary serous and clear cell histologies because of the poor survival and high failure rate in the upper abdomen.


Formation of a fairly stable diazoate intermediate of 5-methyl-2'-deoxycytidine by HNO2 and NO, and its implication to a novel mutation mechanism in CpG site.
The intermediate produced from 5-methyl-2'-deoxycytidine ((5me)dCyd) by HNO2 and NO treatments was isolated and characterized. When 10mM (5me)dCyd was incubated with 100mM NaNO2 at pH 3.7 and 37 degrees C, a previously unidentified product was formed. The product was identified as a diazoate derivative of (5me)dCyd, 1-(beta-D-2'-deoxyribofuranosyl)-5-methyl-2-oxopyrimidine-4-diazoate ((5me)dCyd-diazoate), on the bases of several measurements including LC/MS. The time course of the concentration change of the diazoate showed a characteristic profile of a reaction intermediate, and the steady state concentration was 2.3 microM (0.023% yield). When an aqueous solution of 10mM (5me)dCyd (10 mL) was bubbled by NO at 37 degrees C under aerobic conditions holding the pH around 7.4, the diazoate was also generated. The yield of the diazoate was 0.041 micromol (0.041% yield) at 20 mmol of NO absorption. At physiological pH and temperature (pH 7.4, 37 degrees C), the diazoate was converted to dThd exclusively with a first order rate constant k=9.1x10(-6) x s(-1) (t(1/2)=21 h). These results show that the diazoate is generated as a relatively stable intermediate in the reactions of (5me)dCyd with HNO2 and NO and further suggest that the diazoate can be formed in cellular DNA with biologically relevant doses of HNO2 and NO.


Tubular site of renal sodium retention in ascitic liver cirrhosis evaluated by lithium clearance.
Renal tubular sodium handling was evaluated in 27 non-azotemic cirrhotic patients with ascites and positive sodium balance and in 17 controls after at least 5 days of a constant sodium intake using the lithium clearance as an index of fluid delivery to the distal tubule. Plasma renin activity and plasma aldosterone were also evaluated. Sodium fractional excretion, filtered sodium load, absolute sodium distal delivery, lithium fractional excretion and absolute distal sodium reabsorption were significantly lower in cirrhotics than in controls (0.58 +/- 0.11 vs. 1.29 +/- 0.12%, P less than 0.001; 12529 +/- 677 vs. 15707 +/- 796 microEq min-1 1.73 m-2 BSA, P less than 0.005; 2384 +/- 135.2 vs. 3685 +/- 219.3 microEq min-1 1.73 m-2 BSA, P less than 0.001; 19.5 +/- 1.0 vs. 24.2 +/- 1.3%, P less than 0.01; 2299 +/- 127 vs. 3485 +/- 214 microEq min-1 1.73 m-2 BSA, P less than 0.001, respectively). A correlation was found between lithium clearance and sodium clearance only in cirrhotic patients (r = 0.62; P less than 0.01). Distal sodium reabsorption evaluated as a per cent of filtered sodium load was lower in cirrhotics than in controls (19.1 +/- 1.0 vs. 22.4 +/- 1.2%, P less than 0.05) while distal sodium reabsorption evaluated as a per cent of sodium distal delivery was higher in cirrhotics than in controls (96.7 +/- 0.4 vs. 94.4 +/- 0.5%, P less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)


Supplementation of Freezing Medium with Ginseng Improves Rooster Sperm Quality and Fertility Relative to Free Radicals and Antioxidant Enzymes.
To the best of our knowledge, this study is the first to determine the effect of ginseng as an antioxidant supplement in freezing extenders on the quality of cryopreserved rooster semen. Semen samples were collected from 40 Thai native roosters (Pradu Hang Dum) using the dorso-abdominal massage method and then pooled and divided into five groups according to the concentrations of ginseng supplementation (0, 0.25, 0.50, 0.75, and 1 mg/mL) in a freezing extender. The semen suspensions were loaded into a medium straw and cryopreserved using the liquid nitrogen vapor method. The post-thaw semen was evaluated for sperm quality (sperm motility and membrane integrity), seminal plasma characteristics (lipid peroxidation, superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GPx]), and fertility. The results showed that ginseng extract supplementation at 0.25 mg/mL yielded the highest total motility, progressive motility, and membrane integrity (59.47%, 30.82%, and 48.30%, respectively; p < 0.05) in cryopreserved rooster semen. Higher malondialdehyde concentrations were observed in the control group than in the other groups (p < 0.05). SOD, CAT, and GPx increased compared with those in the control group (p < 0.05). The results showed that the fertility rate with 0.25 mg/mL of ginseng was higher than that of the control group (62.80% vs. 46.28%: p < 0.05). In conclusion, supplementation with 0.25 mg/mL of ginseng is recommended as an alternative component to the freezing extender to improve rooster semen cryopreservation.


Associations between the worksite environment and perceived health culture.
Recognition regarding the importance of health culture in the worksite health promotion literature is growing; however, little empirical evidence exists to guide programmatic or research efforts. The purpose of this study is to examine the relationship between the health supporting structural features of the workplace and perceived health culture. Cross-sectional. Twenty-one worksites with culture data from 2467 employees from Western New York. The Heart Check (HC) and Lifegain Health Culture Audit (LHCA) were used to assess worksite environment and worksite health culture, respectively. Pearson r was used to examine the associations between HC factors and LHCA mean scores. Multiple linear regression was used to predict LHCA mean scores from selected HC factors. Adjusting for age and gender, several significant correlations between HC and LHCA scores were identified, ranging from r = .54 (p < .05) to r = .72 (p < .001). The HC Environmental Structure factor (HCES) and HC Communication factor (HCC) were significantly correlated to LHCA scores (r = .55 and r = .72, respectively; p < .01) but not to each other. A regression model combining HCES and HCC was found to be predictive of LHCA score (R(2) = .69, p < .001). Modifiable components of the worksite environment are strongly and positively related to employee perceptions of the worksite health culture.


Polarity and asymmetric cell division in the control of lymphocyte fate decisions and function.
Polarity is important in several lymphocyte processes including lymphocyte migration, formation of the immunological synapse, and asymmetric cell division (ACD). While lymphocyte migration and immunological synapse formation are relatively well understood, the role of lymphocyte ACD is less clear. Recent advances in measuring polarity enable more robust analyses of asymmetric cell division. Use of these new methods has produced crucial quantification of ACD at precise phases of lymphocyte development and activation. These developments are leading to a better understanding of the drivers of fate choice during lymphocyte activation and provide a context within which to explain the effects of ACD.


Arthroscopic On-Lay Biceps Tenodesis: The Loop-Lock Technique.
The biceps tendon is a common pain generator in the shoulder. Long head of the biceps tendon pathology occurs in a variety of different ways. There are several different treatment options available to address the long head of the biceps. With advances in arthroscopy, all-arthroscopic tenodesis is becoming a more popular choice to address biceps tendon pathology. We describe an all-arthroscopic technique, termed "the loop-lock," for performing a biceps tenodesis.


Regulation of Toll-like receptor signaling by NDP52-mediated selective autophagy is normally inactivated by A20.
Toll-like receptor (TLR) signaling is linked to autophagy that facilitates elimination of intracellular pathogens. However, it is largely unknown whether autophagy controls TLR signaling. Here, we report that poly(I:C) stimulation induces selective autophagic degradation of the TLR adaptor molecule TRIF and the signaling molecule TRAF6, which is revealed by gene silencing of the ubiquitin-editing enzyme A20. This type of autophagy induced formation of autophagosomes and could be suppressed by an autophagy inhibitor and lysosomal inhibitors. However, this autophagy was not associated with canonical autophagic processes, including involvement of Beclin-1 and conversion of LC3-I to LC3-II. Through screening of TRIF-interacting 'autophagy receptors' in human cells, we identified that NDP52 mediated the selective autophagic degradation of TRIF and TRAF6 but not TRAF3. NDP52 was polyubiquitinated by TRAF6 and was involved in aggregation of TRAF6, which may result in the selective degradation. Intriguingly, only under the condition of A20 silencing, NDP52 could effectively suppress poly(I:C)-induced proinflammatory gene expression. Thus, this study clarifies a selective autophagic mechanism mediated by NDP52 that works downstream of TRIF-TRAF6. Furthermore, although A20 is known as a signaling fine-tuner to prevent excess TLR signaling, it paradoxically downregulates the fine-tuning effect of NDP52 on TLR signaling.


Electrodermal lability and simple reaction time.
Subjects identified as electrodermally labile or stabile (n = 10 per group) on the basis of non-specific electrodermal fluctuations and a trials-to-criterion measure of habituation were compared in a simple reaction time (RT) task that employed long, variable foreperiods (from 8 to 19 s). Labiles had significantly faster RTs across all foreperiods, but there were no differences in RT variability between the groups. Phasic heart rate responses recorded concurrently through the foreperiod showed a characteristic pattern of acceleratory and deceleratory activity, which was similar among both labiles and stabiles. The findings indicate that labiles and stabiles show a consistent difference in the speed of processing of a target imperative stimulus, which is not due to differences in response preparation processes. Overall, the results support the use of RT paradigms for evaluating individual differences in information-processing associated with electrodermal lability.


The role of transbronchial lung biopsy in diffuse pulmonary disease.
Forty consecutive patients underwent flexible fiberoptic transbronchial lung biopsy for diagnosis of diffuse nodular or infiltrative lung disease. Biplane fluoroscopic examination with image intensification greatly facilitated accurate placement of the biopsy forceps near the pleura; Specimens of lung parenchyma were obtained for culture and histological study in every case. A pathological diagnosis was correctly established in 34 of 40 patients. Transbronchial biopsy was helpful in the clinical management of an additional 4 patients. Biopsy results were not accurate in 2 patients. No significant morbidity was associated with the procedure. Fiberoptic transbronchial lung biopsy is a safe and useful adjunct to the diagnosis of parenchymal lung disease.


Kinetics of serum glutamate dehydrogenase during short-term alcohol withdrawal.
The behavior of serum glutamate dehydrogenase after alcohol withdrawal was studied in 64 alcoholics admitted to a detoxification center. The enzyme activity decreased in 57 patients by a median of -35.8 percent in 24 h. Thus, the decrease of serum glutamate dehydrogenase after 24 h of ethanol abstinence can serve as a test of alcohol abuse.


The impact of hepatitis B vaccination in a Western country: recall of vaccination and serological status in Australian adults.
Notifications of acute icteric hepatitis B have declined since the introduction of vaccination but it is not clear whether the reservoir of infection and the proportion of adults who remain susceptible have also changed. This has been investigated by evaluation of serological evidence of infection and immunity, patient recall of vaccination and risk factors for exposure to hepatitis B in 2115 adult endoscopy patients in central Sydney. Twenty-one percent were immune, two thirds of these by vaccination. One third of the 440 who recalled "vaccination" were anti-HBs negative. 2.1% of the cohort was HBsAg positive and of these 31% (14/45) were viraemic. Amongst epidemiological risk groups recommended for vaccination, multivariate analysis showed that health care workers (odd ratio, OR = 5.35) and patients diagnosed with hepatitis A (OR = 2.6) or hepatitis C (OR = 2.1) were 5.35, 2.6 and 2.1 times more likely to be immunised, respectively. The great majority of immigrants from high prevalence countries, and of patients reporting other known risks for hepatitis B exposure remain susceptible.


Isoquercetin upregulates antioxidant genes, suppresses inflammatory cytokines and regulates AMPK pathway in streptozotocin-induced diabetic rats.
Lifestyle and genetic factors contribute to the initiation of oxidative stress and inflammation in diabetes mellitus (DM). Oxidative stress and lipid peroxidation worked in an orchestrated manner and reported to be strongly associated with the formation of the hyperlipidemic condition in DM patients. Isoquercetin, a bioactive constituent isolated from guava leaves has attracted considerable attention because of its antidiabetic activity. The antidiabetic activity of guava leaves may be due to the presence of isoquercetin at a significant level. However, how isoquercetin regulates different pathways in DM is insufficiently studied. We have selected versatile regulators of oxidative stress and inflammatory pathways to fully analyze if isoquercetin effectively modulated the genes of these pathways. At the end of our experimental duration, rats were dissected and analyzed for the oxidative stress, lipid peroxidation, inflammatory and lipid markers. The nuclear factor erythroid 2-related factor 2 (Nrf2) pathway is believed to be the key regulator of expression of various antioxidant enzyme genes and it is directly or indirectly related to nuclear factor Kappa- B (NF-kB) and AMP-activated protein kinase (AMPK) pathways. Therefore, we tend to study the effects of STZ on Nrf2, NF-kB and AMPK pathway and how the isoquercetin treatment performs at a molecular level to overcome the burden of DM. The results of our study provided convincing evidence of significant pharmacological properties of isoquercetin in context of its ability to inhibit the oxidative stress elicited by the STZ through generation of the free radicals and regulation of the expression of Nrf2 pathway-associated proteins and genes and it also reduced the burden of hyperlipidemia and inflammation. By taking the above results into consideration isoquercetin can be studied further to elucidate its antidiabetic effects at various levels.


A Phase Ib, Open-label Study Evaluating the Safety and Efficacy of Ipatasertib plus Rucaparib in Patients with Metastatic Castration-resistant Prostate Cancer.
To report the safety and efficacy of ipatasertib (AKT inhibitor) combined with rucaparib (PARP inhibitor) in patients with metastatic castration-resistant prostate cancer (mCRPC) previously treated with second-generation androgen receptor inhibitors. In this two-part phase Ib trial (NCT03840200), patients with advanced prostate, breast, or ovarian cancer received ipatasertib (300 or 400 mg daily) plus rucaparib (400 or 600 mg twice daily) to assess safety and identify a recommended phase II dose (RP2D). A part 1 dose-escalation phase was followed by a part 2 dose-expansion phase in which only patients with mCRPC received the RP2D. The primary efficacy endpoint was prostate-specific antigen (PSA) response (≥50% reduction) in patients with mCRPC. Patients were not selected on the basis of tumor mutational status. Fifty-one patients were enrolled (part 1 = 21; part 2 = 30). Ipatasertib 400 mg daily plus rucaparib 400 mg twice daily was the selected RP2D, received by 37 patients with mCRPC. Grade 3/4 adverse events occurred in 46% (17/37) of patients, with one grade 4 adverse event (anemia, deemed related to rucaparib) and no deaths. Adverse events leading to treatment modification occurred in 70% (26/37). The PSA response rate was 26% (9/35), and the objective response rate per Response Criteria in Solid Tumors (RECIST) 1.1 was 10% (2/21). Median radiographic progression-free survival per Prostate Cancer Working Group 3 criteria was 5.8 months [95% confidence interval (CI), 4.0-8.1], and median overall survival was 13.3 months (95% CI, 10.9-not evaluable). Ipatasertib plus rucaparib was manageable with dose modification but did not demonstrate synergistic or additive antitumor activity in previously treated patients with mCRPC.


Structure-function analysis of GNIP, the glycogenin-interacting protein.
Glycogenin is a self-glucosylating protein that initiates glycogen biosynthesis. We recently identified a family of proteins, GNIPs, that interact with glycogenin and stimulate its self-glucosylating activity [J. Biol. Chem. 277 (2002) 19331]. The GNIP gene (also called TRIM7) encodes at least four distinct isoforms of GNIP, three of which (GNIP1, GNIP2, and GNIP3) have in common a COOH-terminal B30.2 domain and predicted coiled-coil regions. Based on Western blot analysis, the GNIP1 protein is widely distributed in tissues. From analysis of a series of deletion mutants of GNIP2 using the yeast two-hybrid system, the B30.2 domain was found to be responsible for the interaction with glycogenin. A truncated form of recombinant GNIP2, lacking the NH2-terminal coiled-coil region, was cross-linked to glycogenin by glutaraldehyde treatment, supporting the idea that the B30.2 domain was sufficient for the interaction. In the course of this study, GNIP2 was also found to interact with itself, via the coiled-coil domain. Heterologous interactions between GNIP1 and GNIP2 were also detected. Since glycogenin is also a dimer, higher order multimeric complexes between glycogenin and GNIPs would be possible.


Dendritic activities of spinal motoneurones in pigs and rabbits enhanced through chronic stimulation of a dorsal root.
1. The L6 or L7 dorsal root in 7-10-day-old pigs was chronically stimulated for 3 or 4 h a day for 4 days/week. The stimuli were 50 or 500 Hz pulse trains delivered at 0.5/s. The induced movements were limited to the hip and hind leg on the stimulated side without any sign of pain. There was evidence that not only the monosynaptic, but also polysynaptic pathways to the motoneurones were chronically stimulated. After 2-6 days of chronic stimulation, all pigs were studied in acute experiments either under pentobarbitone (Nembutal) anaesthesia or in the decerebrated state. Normal (unstimulated) 15-28-day-old pigs were also studied in acute experiments under similar conditions, as the controls. 2. In thirty-three out of fifty-six motoneurones of normal pigs there were all-or-none potentials of 2-6 ms duration and 1-5 mV amplitude. These have been named D-spikes. They occurred in response to tibial nerve and/or dorsal root stimulation and/or spontaneously. They could be abolished by injection of a hyperpolarizing current. These results show that D-spikes are different from any of the well-established all-or-none events of the motoneurone, i.e. the spikes of the axon, initial segment, soma and unitary excitatory post-synaptic potentials. Several kinds of D-spikes could exist in the same motoneurone, as judged from their wave forms and thresholds. This argues for the dendrites as the sites of origin for D-spikes, because only the dendrites can have multiple sites for spike generation. D-spikes were shown to summate to form a larger depolarization. 3. In pigs chronically stimulated with 500 Hz pulses seventy-five motoneurones were studied in acute experiments. In sixty-one motoneurones D-spikes were observed. This indicated that D-spike occurrence was enhanced in the chronically stimulated motoneurones as compared with controls. 4. Characteristic of the chronically stimulated motoneurone was the presence of a depolarization of all-or-none nature which was more than 10 ms in duration and 2-12 mV in amplitude. The depolarization is referred to as the D-wave. The D-wave was similar in properties to the D-spike described above, except that it was of longer duration. Consequently, it is assumed that the D-wave is summed D-spikes. D-waves were observed in nineteen motoneurones.(ABSTRACT TRUNCATED AT 400 WORDS)


BmTI antigens induce a bovine protective immune response against Boophilus microplus tick.
Boophilus microplus trypsin inhibitors (BmTIs) present in larvae were preliminarily characterized as active proteins, approximately 10-18 kDa, by SDS-PAGE. BmTIs showed trypsin inhibitory activity on reverse zymography containing gelatin (0.03%) and also inhibited others serine proteinases (human neutrophil elastase and human plasma kallikrein). Bos indicus, Nelore breed calves, previously sensitized with BmTIs and challenged with tick larvae (20,000 larvae/animal), showed 72.8% efficacy to interfere in tick development with 69.7% and 71.3% reduction of both tick number and egg weight, respectively. Cattle BmTls antiserum titer was approximately 1:8000. The maximum level of BmTls antibody production was detected 40 days after the first immunization by ELISA. Our preliminary results suggest that B. microplus serine proteinase inhibitors may play a role in the tick larvae fixation and feeding processes. Therefore, the development of antibodies against BmTIs might impair the normal parasitism.


Choline increases acetylcholine release and protects against the stimulation-induced decrease in phosphatide levels within membranes of rat corpus striatum.
This study examined the possibility that membrane phospholipids might be a source of choline used for acetylcholine (ACh) synthesis. Slices of rat striatum or cerebellum were superfused with a choline-free or choline-containing (10, 20 or 40 microM) physiological solution with eserine, for alternating 20 min periods of rest or electrical stimulation. Superfusion media were assayed for choline and ACh, and slice samples taken before and after stimulation were assayed for choline, ACh, various phospholipids, protein and DNA. The striatal slices were able to sustain the stimulation-induced release of ACh, releasing a total of about 3 times their initial ACh contents during the 8 periods of stimulation and rest. During these 8 cycles, 885 pmol/micrograms DNA free choline was released from the slices into the medium, an amount about 45-fold higher than the initial or final free choline levels in the slices. Although repeated stimulation of the striatal slices failed to affect tissue levels of free choline or of ACh, this treatment did cause significant, dose-related (i.e., number of stimulation periods) stoichiometric decreases in tissue levels of phosphatidylcholine (PC) and of the other major phospholipids; tissue protein levels also declined significantly. Addition of exogenous choline to the superfusion medium produced dose-related increases in resting and evoked ACh release. The choline also fully protected the striatal slices from phospholipid depletion for as many as 6 stimulation periods. Cerebellar slices liberated large amounts of free choline into the medium but did not release measurable quantities of ACh; their phospholipid and protein levels did not decline with electrical stimulation. These data show that membrane phospholipids constitute a reservoir of free choline that can be used for ACh synthesis. When free choline is in short supply, ACh synthesis and release are sustained at the expense of this reservoir. The consequent reduction in membrane PC apparently is associated with a depletion of cellular membrane. The use of free choline by cholinergic neurons for two purposes, the syntheses of both ACh and membrane phospholipids, may thus impart vulnerability to them in situations where the supply of free choline is less than that needed for acetylation.


Pharmacokinetics of a single orally administered therapeutic dosage of cyclosporine A in healthy cats.
This study aimed to determine a pharmacokinetic profile for a single dosage of cyclosporine A (CsA) clinically used for immunosuppression in cats. Blood-CsA-concentrations were measured before and 1, 2, 4, 6, 8, 12 and 24 h after oral administration of 7 mg/kg body weight (BW) CsA (Atopica® oral solution) to 8 healthy adult cats using high-performance liquid chromatography coupled to mass spectrometry. Pharmacokinetic parameters were calculated using WinNonLin software based on a 1-compartment-model. The median maximum plasma-concentration of 1466 ng/ml (530-2235 ng/ml; minimum-maximum) was reached after 2.0 h (1.0-4.7 h). The area under the curve was 12,568 h x ng/ml (5732-20,820 h x ng/ml) and the apparent total clearance of the drug from plasma was 557 ml/h/kg (336-1221 ml/h/kg). Half-life of absorption into the central compartment was 0.6 h (0.4-2.6 h), half-life of elimination from the central compartment was 4.6 h (1.4-7.5 h).


Erratum to link between environmental air pollution and allergic asthma: East meets West.
[This corrects the article on p. 14 in vol. 7, PMID: 25694814.].


Structure and catalytic mechanism of horseradish peroxidase. Regiospecific meso alkylation of the prosthetic heme group by alkylhydrazines.
Horseradish peroxidase is inactivated in a time-, H2O2-, and concentration-dependent manner by phenylethyl-, ethyl-, and methylhydrazine. The pseudo- first order kinetic constants for these inactivation reactions at pH 7 are: phenylethyl (KI = 115 microM, kinact = 1.5 min-1, partition ratio = 11), ethyl (KI = 145 microM, kinact = 0.08 min-1, partition ratio = 32), and methyl (KI = 3000 microM, kinact = 0.12 min-1, partition ratio = 80). At pH 5, the constants for the phenylethyl reaction change to KI = 1540 microM and kinact = 0.86 min-1. A transient absorbance at approximately 830 nm, suggestive of an isoporphyrin intermediate, is seen during these reactions. The prosthetic heme is converted by each of the three alkylhydrazines into the corresponding delta-meso-alkylated heme. Complete inactivation of the enzymes by methyl-, ethyl-, and phenylethylhydrazine is associated with alkylation of 60-70, 70, and 90%, respectively, of the prosthetic heme groups. The absence of N-alkylation and the high specificity for the delta-meso position, even with agents as small as methylhydrazine, strengthen the proposal that electron abstraction is mediated by the heme edge rather than the ferryl oxygen of horseradish peroxidase.


Structure-function subsystem models of female and male forebrain networks integrating cognition, affect, behavior, and bodily functions.
The forebrain is the first of three primary vertebrate brain subdivisions. Macrolevel network analysis in a mammal (rat) revealed that the 466 gray matter regions composing the right and left sides of the forebrain are interconnected by 35,738 axonal connections forming a large set of overlapping, hierarchically arranged subsystems. This hierarchy is bilaterally symmetrical and sexually dimorphic, and it was used to create a structure-function conceptual model of intraforebrain network organization. Two mirror image top-level subsystems are presumably the most fundamental ontogenetically and phylogenetically. They essentially form the right and left forebrain halves and are relatively weakly interconnected. Each top-level subsystem in turn has two second-level subsystems. A ventromedial subsystem includes the medial forebrain bundle, functionally coordinating instinctive survival behaviors with appropriate physiological responses and affect. This subsystem has 26/24 (female/male) lowest-level subsystems, all using a combination of glutamate and GABA as neurotransmitters. In contrast, a dorsolateral subsystem includes the lateral forebrain bundle, functionally mediating voluntary behavior and cognition. This subsystem has 20 lowest-level subsystems, and all but 4 use glutamate exclusively for their macroconnections; no forebrain subsystems are exclusively GABAergic. Bottom-up subsystem analysis is a powerful engine for generating testable hypotheses about mechanistic explanations of brain function, behavior, and mind based on underlying circuit organization. Targeted computational (virtual) lesioning of specific regions of interest associated with Alzheimer's disease, clinical depression, and other disorders may begin to clarify how the effects spread through the entire forebrain network model.


Evolution of dietary diversity and a starvation driven cross-diffusion system as its singular limit.
We rigorously prove the passage from a Lotka-Volterra reaction-diffusion system towards a cross-diffusion system at the fast reaction limit. The system models a competition of two species, where one species has a more diverse diet than the other. The resulting limit gives a cross-diffusion system of a starvation driven type. We investigate the linear stability of homogeneous equilibria of those systems and rule out the possibility of cross-diffusion induced instability (Turing instability). Numerical simulations are included which are compatible with the theoretical results.


Saccadic motor planning by integrating visual information and pre-information on neural dynamic fields.
A functional model of target selection in the saccadic system is presented, incorporating elements of visual processing, motor planning, and motor control. We address the integration of visual information with pre-information, which is provided by manipulating the probability that a target appears at a certain location. This integration is achieved within a dynamic representation of planned eye movement which is modeled through distributions of activation on a topographic field. Visual input evokes activation, which is also constrained by lateral interaction within the field and by preshaping input representing pre-information. The model describes target selection observable in paradigms in which visual goals are presented at more than one location. Specifically, we model the transition from averaging, where endpoints of first saccades fall between two visual target locations, to decision making, where endpoints of first saccades fall accurately onto one of two simultaneously presented visual targets. We make predictions about how metrical biases of first saccades are induced by pre-information about target locations acquired by learning. When coupled to a motor control stage, activation dynamics on the planning level contribute to stabilizing gaze under fixation conditions. The neurophysiological relevance of our functional model is discussed.