Dissection of the genetic architecture of three seed-quality traits and consequences for breeding in Brassica napus.
Genome-wide association studies (GWASs) combining high-throughput genome resequencing and phenotyping can accelerate the dissection of genetic architecture and identification of genes for plant complex traits. In this study, we developed a rapeseed genomic variation map consisting of 4 542 011 SNPs and 628 666 INDELs. GWAS was performed for three seed-quality traits, including erucic acid content (EAC), glucosinolate content (GSC) and seed oil content (SOC) using 3.82 million polymorphisms in an association panel. Six, 49 and 17 loci were detected to be associated with EAC, GSC and SOC in multiple environments, respectively. The mean total contribution of these loci in each environment was 94.1% for EAC and 87.9% for GSC, notably higher than that for SOC (40.1%). A high correlation was observed between phenotypic variance and number of favourable alleles for associated loci, which will contribute to breeding improvement by pyramiding these loci. Furthermore, candidate genes were detected underlying associated loci, based on functional polymorphisms in gene regions where sequence variation was found to correlate with phenotypic variation. Our approach was validated by detection of well-characterized FAE1 genes at each of two major loci for EAC on chromosomes A8 and C3, along with MYB28 genes at each of three major loci for GSC on chromosomes A9, C2 and C9. Four novel candidate genes were detected by correlation between GSC and SOC and observed sequence variation, respectively. This study provides insights into the genetic architecture of three seed-quality traits, which would be useful for genetic improvement of B. napus.


Hyponatremia, heart failure, and the role of tolvaptan.
Hyponatremia-usually defined by serum sodium < 135 mEq/L-is common in heart failure (HF); it remains unclear whether it worsens HF or is merely a marker of more severe disease. Hyponatremia may develop from causes besides HF and symptoms may be mistakenly attributed to HF. Hyponatremia correction may be required for optimal HF management in some cases, and it can prevent neurologic complications. Symptoms, volume status, and onset timing determine treatment, which should correct serum sodium in a controlled manner. Arginine vasopressin is elevated in hypervolemic/euvolemic hyponatremia, favoring water reabsorption despite low serum osmolality. The oral selective V(2)-receptor antagonist tolvaptan blocks arginine vasopressin effects in the renal collecting ducts, promoting aquaresis without increasing sodium/potassium excretion. In clinical trials, tolvaptan significantly increased serum sodium in patients with euvolemic/hypervolemic hyponatremia, including HF. When added to conventional HF treatment, tolvaptan produced early symptomatic benefit, without long-term improvement in an HF population consisting primarily of normonatremic patients. Tolvaptan is approved for treatment of clinically significant hypervolemic/euvolemic hyponatremia (serum sodium < 125 mEq/L or less marked symptomatic hyponatremia that has resisted correction with fluid restriction), but not HF without hyponatremia.


Therapeutic options for human brucellosis.
Worldwide, human brucellosis is the most common zoonotic disease and it has gained increasing interest because of the potential use of Brucella as a biological weapon. Monotherapy for brucellosis is associated with a high relapse rate and dual therapy in different combinations is more efficacious. The combination regimen of intramuscular streptomycin with an oral tetracycline resulted in fewer relapses than the doxycycline-rifampin combination in meta-analysis and prospective studies, although the use of doxycycline and rifampin is a reasonable choice in certain conditions. Longer duration and triple antimicrobial therapy appear to improve outcome and prevent relapses, especially in patients with focal disease. Recently, the use of gentamicin-loaded microparticles and the use of new antibiotics, such as tigecycline, may hold future promise. In addition, there are a few studies of the enhanced effect of immune response stimulators, such as levimasole and IFN-2, in the treatment of brucellosis. The development of an effective subcellular Brucella vaccine would be an important step forward to curtail the disease. However, currently and for the near future, only the control of animal disease is possible using vaccine strategies.


Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells.
Erythropoietin (Epo) is a glycoprotein hormone that regulates erythroid development and interacts with surface receptors on developing erythroid cells. In this laboratory, a cell system with a relatively pure population of erythroid cells that respond to Epo has been developed. Immature erythroid cells are obtained from the spleens of mice infected with the anemia strain of Friend virus. The binding of 125I-labeled Epo (125I-Epo) to plasma membranes from these cells was studied in this investigation. 125I-Epo binding reached equilibrium within 20 min at 37 degrees C. Twenty percent of the receptors bound 125I-Epo with a Kd of 0.08 X 10(-9) M, while the remaining receptors bound the hormone with a Kd of 0.6 X 10(-9) M. In this study, a receptor for Epo was identified by cross-linking 125I-Epo to the receptor in intact cells and plasma membrane preparations using disuccinimidyl suberate. Polyacrylamide gel electrophoresis revealed two labeled bands of 100 and 85 kDa. The 85-kDa band was more heavily labeled (65%) than the 100-kDa band. Both bands were equally decreased when increasing amounts of unlabeled Epo were included in the binding mixture, indicating a specific interaction of 125I-Epo with the receptor.


Differential association between MAOA, ADHD and neuropsychological functioning in boys and girls.
Attention-deficit/hyperactivity disorder (ADHD) is more common in boys than in girls. It has been hypothesized that this sex difference might be related to genes on the X-chromosome, like Monoamine Oxidase A (MAOA). Almost all studies on the role of MAOA in ADHD have focused predominantly on boys, making it unknown whether MAOA also has an effect on ADHD in girls, and few studies have investigated the relationship between MAOA and neuropsychological functioning, yet this may provide insight into the pathways leading from genotype to phenotype. The current study set out to examine the relationship between MAOA, ADHD, and neuropsychological functioning in both boys (265 boys with ADHD and 89 male non-affected siblings) and girls (85 girls with ADHD and 106 female non-affected siblings). A haplotype was used based on three single nucleotide polymorphisms (SNPs) (rs12843268, rs3027400, and rs1137070). Two haplotypes (GGC and ATT) captured 97% of the genetic variance in the investigated MAOA SNPs. The ATT haplotype was more common in non-affected siblings (P = 0.025), conferring a protective effect for ADHD in both boys and girls. The target and direction of the MAOA effect on neuropsychological functioning was different in boys and girls: The ATT haplotype was associated with poorer motor control in boys (P = 0.002), but with better visuo-spatial working memory in girls (P = 0.01). These findings suggest that the genetic and neuropsychological mechanisms underlying ADHD may be different in boys and girls and underline the importance of taking into account sex effects when studying ADHD.


[Mutagenic action of O-methlhydroxylamide on transforming DNA].
The mutagenic effect of O-methylhydroxylamine (OMHA) on transforming DNA of Bacillus subtilis was studied. In accordance with the earlier reported chemical and functional data, the mutagenic effect was observed at 4.5 and 6.0 pH. An increase in pH caused a decrease in the rate of mutagenesis, though the maximal level of mutagenesis was equal at both values of pH. The results obtained with recipients defective in the system of UV-repair revealed that both products of reaction of OMHA with the cytosine-base of DNA, N4-metoxycytidine and N4-metoxy-6-metoxyamino-5,6-dihydrocytidine, are effectively eliminated through the system of UV repair.


Ultrasound-Induced Cascade Amplification in a Mechanoluminescent Nanotransducer for Enhanced Sono-Optogenetic Deep Brain Stimulation.
Remote and genetically targeted neuromodulation in the deep brain is important for understanding and treatment of neurological diseases. Ultrasound-triggered mechanoluminescent technology offers a promising approach for achieving remote and genetically targeted brain modulation. However, its application has thus far been limited to shallow brain depths due to challenges related to low sonochemical reaction efficiency and restricted photon yields. Here we report a cascaded mechanoluminescent nanotransducer to achieve efficient light emission upon ultrasound stimulation. As a result, blue light was generated under ultrasound stimulation with a subsecond response latency. Leveraging the high energy transfer efficiency of focused ultrasound in brain tissue and the high sensitivity to ultrasound of these mechanoluminescent nanotransducers, we are able to show efficient photon delivery and activation of ChR2-expressing neurons in both the superficial motor cortex and deep ventral tegmental area after intracranial injection. Our liposome nanotransducers enable minimally invasive deep brain stimulation for behavioral control in animals via a flexible, mechanoluminescent sono-optogenetic system.


The potential role of ectomycorrhizal fungi in determining Douglas-fir resistance to defoliation by the western spruce budworm (Lepidoptera: Tortricidae).
There is phenotypic variation among individual trees of interior Douglas-fir (Pseudotsuga menziesii var. glauca [Beissn.] Franco) in their resistance to defoliation by the western spruce budworm (Choristoneura occidentalis Freeman). We evaluated the potential role of ectomycorrhizal fungi in determining this resistance using half-sib seedlings derived from parent trees that are resistant versus susceptible to budworm defoliation in the field. The seedlings were inoculated with Laccaria bicolor ectomycorrhizal fungi, fertilized, or untreated. Approximately 48 d after treatment, late-instar larvae from a nondiapausing laboratory colony of C. occidentalis were allowed to feed on pairs of resistant versus susceptible seedlings for 1 wk. Chemical analyses of current-year shoots for nitrogen (N), phosphorus (P), magnesium (Mg), and zinc (Zn) indicated that the fungus increased foliar concentrations of P and Mg in resistant seedlings, but it did not increase their growth rate. However, L. bicolor had no effect on foliar concentrations of P or Mg in susceptible seedlings, even though seedling growth rates increased slightly in response to the inoculation. L. bicolor had no effect on foliar levels of N or Zn in any of the seedlings. As expected, fertilization increased levels of N and P in the foliage of both resistant and susceptible seedlings, but it did not affect levels of Mg and Zn. Surprisingly, the fertilizer treatment had no effect on seedling growth rates. Despite these differences, late-instar budworms showed no feeding preference among untreated, mycorrhizal, or fertilized seedlings. The fact that seedlings from resistant versus susceptible Douglas-firs responded differently to the L. bicolor treatment lends preliminary support to the hypothesis that ecotmycorrhizae might play a role in Douglas-fir resistance to damage from the western spruce budworm. Finally, differences in foliar concentrations of N and P among untreated seedlings from different maternal trees suggested that foliar nutritional chemistry is influenced by the tree's genotype.


Pathways for Ca2+ efflux in heart and liver mitochondria.
1. Two processes of Ruthenium Red-insensitive Ca2+ efflux exist in liver and in heart mitochondria: one Na+-independent, and another Na+-dependent. The processes attain maximal rates of 1.4 and 3.0 nmol of Ca2+.min-1.mg-1 for the Na+-dependent and 1.2 and 2.0 nmol of Ca2+.min-1.mg-1 for the Na+-independent, in liver and heart mitochondria, respectively. 2. The Na+-dependent pathway is inhibited, both in heart and in liver mitochondria, by the Ca2+ antagonist diltiazem with a Ki of 4 microM. The Na+-independent pathway is inhibited by diltiazem with a Ki of 250 microM in liver mitochondria, while it behaves as almost insensitive to diltiazem in heart mitochondria. 3. Stretching of the mitochondrial inner membrane in hypo-osmotic media results in activation of the Na+-independent pathway both in liver and in heart mitochondria. 4. Both in heart and liver mitochondria the Na+-independent pathway is insensitive to variations of medium pH around physiological values, while the Na+-dependent pathway is markedly stimulated parallel with acidification of the medium. The pH-activated, Na+-dependent pathway maintains the diltiazem sensitivity. 5. In heart mitochondria, the Na+-dependent pathway is non-competitively inhibited by Mg2+ with a Ki of 0.27 mM, while the Na+-independent pathway is less affected; similarly, in liver mitochondria Mg2+ inhibits the Na+-dependent pathway more than it does the Na+-independent pathway. In the presence of physiological concentrations of Na+, Ca2+ and Mg2+, the Na+-independent and the Na+-dependent pathways operate at rates, respectively, of 0.5 and 1.0 nmol of Ca2+.min-1.mg-1 in heart mitochondria and 0.9 and 0.2 nmol of Ca2+.min-1.mg-1 in liver mitochondria. It is concluded that both heart and liver mitochondria possess two independent pathways for Ca2+ efflux operating at comparable rates.


Depression and anxiety-related disorders and suicide among Swedish male elite football players: a nationwide cohort study.
To assess whether male elite football players, during and after their active career, were at increased risk of depression and anxiety-related disorders and suicide, as compared with the general male population. We included male football players active in the Swedish top division 1924-2019 and general male population (matched to football players based on age and region of residence) aged <65 years in 1997. Using nationwide registers, we followed the football players from their first season in the top division (or the date of their first registered residency in Sweden) or 1 January 1997, and compared the risk of depression and anxiety-related disorders (captured through diagnoses from hospital admissions and outpatient visits, and use of prescription drugs) among football players versus controls. In a secondary analysis using data from death certificates, we compared the risk of suicide between football players and general population males who were alive in 1969 (when cause of death became available) . During follow-up through 31 December 2020, 504 (13.6%) of 3719 football players and 7455 (22.3%) of 33 425 general population males had a depression or anxiety-related disorder. In analyses accounting for age, region of residence and calendar time, the risk of anxiety and depression-related disorders was lower among football players versus general population males (HR 0.61, 95% CI 0.55 to 0.66). The protective association was attenuated with increasing age, and from around age 70 years the risk was similar in the two groups. The risk of suicide was lower among football players versus general population males (HR 0.48, 95% CI 0.32 to 0.72). In this nationwide cohort study in Sweden, elite male football players had a lower risk of depression and anxiety-related disorders and suicide as compared with the general population.


Approved or disregarded? Exploring arenas for narrative relations in geriatric care.
The use of narration in healthcare has been accentuated as a response to the requested shift towards person-centred care. The notion of narrative relations refers to a process of involving several people in mutual and ongoing narrative exchange. This study aimed to explore how and where narrative relations may be adopted and enacted in everyday healthcare practice. The study has a qualitative, explorative design. Seven interprofessional focus group discussions with healthcare staff were prompted by vignettes. A multidisciplinary team of healthcare staff (n = 31) were recruited on a geriatric ward. Data were analysed using a constant comparative method. A core theme shows how narrative relations are adopted and enacted both as part of an approved practice-the work procedures commonly approved as part of healthcare, and as a disregarded practice where covert but important narrative relations take place to support fundamental qualities of healthcare. Moreover, the findings consider arenas of healthcare practice where approved or disregarded practices are enacted in the clinic frontstage and the clinic backstage. Narrative relations may take place in different arenas of healthcare practice yet simultaneously become a cohesive force interconnecting those arenas and uphold continuity. Impeded narrative relations in one arena may have unintended consequences in another.


[REHABILITATION OF PATIENTS WITH URIC ACID AND MIXED UROLITHIASIS].
This paper presents evaluation of rehabilitation effectiveness of 113 patients aged 19-78 years diagnosed as having mixed and uric acid stones. 88 patients with uric acid stones 0,5-2,5 cm were assigned to group 1 and treated with Trometamol N parenteral litholis and a complex metaprophilaxis by dietary supplements Prolit and Urisan. 25 patients with mixed stones 1-3,5 cm were allocated to group 2 and treated with external shock wave lithotripsy and complex metaprophilaxis by dietary supplements Prolit Super Septo and Urisan. Positive results were achieved in all the patients. In the patients of group 1 the stones were completely dissolved. In group 2 treatment resulted in stone disintegration and clearance of small fragments 0,4 cm after partial dissolution.


Glycohemoglobin A1c: A promising screening tool in gestational diabetes mellitus.
Current screening tests for gestational diabetes mellitus (GDM) are inconvenient. Therefore, alternative screening tests for GDM are desirable. The use of glycohemoglobin A1c (HbA1c) in screening for GDM remains controversial. We undertook this study to evaluate the utility of HbA1c in screening for GDM. Retrospective study in a tertiary teaching hospital. Laboratory records were reviewed to identify pregnant women who underwent both oral glucose tolerance test (OGTT) and HbA1c measurements over a 16-months period. The association of OGTT with HbA1c was evaluated. Data were collected using SPSS software. Comparisons of the means and calculations of sensitivities were performed. Of 145 eligible patients, 124 had GDM and 21 patients did not, per OGTT. The percentages of patients with HbA1c values (reference range of 4.8%-6.0%) equal to or above sequential cut-point values of 5.0%, 5.5%, 6.0%, 6.5% and 7.0% (i.e., sensitivity values) were 100%, 98.4%, 87.1%, 62.9% and 39.5%, respectively. The mean HbA1c of the patients with GDM was 6.9 + 0.8% compared to 6.4 + 0.6% for those without GDM (P< 0.006). At an arbitrary cut-off value of 6.0% (the upper limit of normal), HbA1c would have picked up 87.1% of patients with GDM. This study suggests that HbA1c is a reasonably sensitive screening measure of GDM in this high-risk population. Acknowledging limitations resulting from the study design, further prospective studies are warranted to verify this conclusion, and to evaluate the specificity of HbA1c as a screening test for GDM.


Withholding negative feedback: is it about protecting the self or protecting others?
The reluctance to deliver negative feedback to someone's face is widely documented. This research disentangles the extent to which this reluctance is motivated by a desire to protect the self as opposed to others. Participants assessed an essay written by someone with high, medium, or low self-esteem. Assessment of the essay was most positive when the feedback was to be provided face-to-face, less positive when delivered anonymously, and least positive when it was not required to be delivered at all. This effect only emerged among participants low in self-liking (but was unrelated to self-competency). The self-esteem of the essay writer had no effect on evaluations. The data lend support for a self-protection motive and modest support for an other-protection motive.


Beta-adrenergic upregulation of the Na(+)-K(+)-2Cl- cotransporter in rat parotid acinar cells.
We used the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6')-carboxyfluorescein to monitor the recovery of the intracellular pH (pHi) of rat parotid acini from an NH4(+)-induced alkaline load. This recovery was markedly inhibited by the loop diuretic bumetanide and by Cl- removal, indicating that it is largely due to NH4+ entry via the basolateral Na(+)-K(+)-2Cl- cotransporter. The rate of recovery of pHi was enhanced threefold by pretreatment (37.5 s) with isoproterenol (K1/2 = 21.5 nM) or norepinephrine (in the presence of phentolamine), and blocked by the beta 1-specific antagonist atenolol, indicating an upregulation of cotransport activity by beta 1-adrenergic stimulation. The effect of isoproterenol was prevented by protein kinase inhibitors and mimicked by cAMP analogues, and by maneuvers known to increase cytosolic cAMP levels in these cells, consistent with the involvement of protein kinase A. Physiologically, such an upregulation of the acinar Na(+)-K(+)-2Cl- cotransporter would lead to an increase in acinar chloride uptake across the basolateral membrane, and consequently, an increase in overall chloride and fluid secretion. Prevention of this upregulation by beta-blockers and possibly by other commonly used clinical agents may account for the dry mouth and dry eyes experienced by some patients taking these medications.


Transjugular intrahepatic portosystemic shunt. Experiences at a liver transplantation center.
Transjugular intrahepatic portosystemic shunt (TIPS) placement is an established therapy for portal hypertension that leads to variceal bleeding or refractory ascites. We present experiences of the role of TIPS at a liver transplantation center. One hundred and ten patients were referred to the Radiological Department for TIPS placement. One of the 110 patients had recurrent cirrhosis after liver transplantation with refractory ascites. Function of the TIPS was controlled with Doppler US at 1 day, 1 week, 4 weeks and 4 months after TIPS placement and subsequently every 3 months. Shunt insufficiency was supposed when the blood flow velocity within the stent tract was under 50 cm/s and was an indication for TIPS revision. TIPS was placed in 101 patients. After TIPS placement, 10 patients underwent liver transplantation. While waiting for the new liver, none of them developed variceal rebleeding, ascites or other complications of portal hypertension. Two of the 101 patients had episodes of rebleeding. The frequency of patients undergoing TIPS revision within the first year after TIPS placement was 67.5%, within the second year 38.0% and within the third year 24.4%. The revisions led to sufficient reduction of the portosystemic pressure gradient. In some liver transplant candidates, TIPS can be useful in minimizing the risk of complications of portal hypertension during the waiting time for a liver transplantation. TIPS can be monitored by Doppler US and revised if occlusion occurs.


Challenges of applying a comprehensive model of intervention fidelity.
Applying a comprehensive model of fidelity to interventions delivered by information and communication technologies (ICTs) has multiple challenges. Fidelity must be considered in the design, implementation, evaluation, and reporting of the intervention. The fidelity strategies must address the unique aspects of the technology, including training providers to instruct participants to use the technology and provide standardized feedback, rather than delivering the intervention in person. Other challenges include the nonspecific effects resulting from participants accessing unintended content in interventions using the Internet. ICT allows participant receipt and enactment of intervention skills to be assessed by electronic evidence, rather than in-person observation. Interventions delivered by ICT are unique, and there is less control of participant interaction with various electronic components. Monitoring participant use and providing standardized feedback for receipt and enactment of intervention skills are key to ensuring fidelity. The final challenges involve evaluating and reporting fidelity of the intervention.


Impact of CMV PCR Blips in Recipients of Solid Organ and Hematopoietic Stem Cell Transplantation.
Viral blips reflecting polymerase chain reaction (PCR) artefacts or transient low-level replication are well described in the human immunodeficiency virus setting. However, the epidemiology of such blips in transplant recipients screened for cytomegalovirus (CMV) with PCR remains uncertain and was investigated in a cohort of solid organ and hematopoietic stem cell recipients. Eligible recipients had known donor/recipient CMV IgG serostatus, and 3 CMV PCRs ≥. The CMV PCR triplicates (3 consecutive CMV PCRs) were defined; the first CMV PCR was always negative, and the time between the second and third samples was 7 days ≤. A positive second but negative third sample represented a blip. Odds ratio (OR) for factors associated with a triplicate being a blip was estimated by binomial regression adjusted for repeated measurements. Whether blips affected the hazard ratio (HR) for subsequent CMV infection was determined with a Cox model. 851 recipients generated 3883 CMV PCR triplicates. The OR of a triplicate representing a blip decreased with increasing viral load of the second sample (vs 273 IU/mL; >273-910 IU/mL: odds ratio [OR], 0.2; 95% confidence interval [CI], 0.1-0.5; >910 IU/mL: OR, 0.08; 95% CI, 0.02-0.2; P ≤ 0.0002) and increased with intermediary-/low-risk serostatus (vs high risk) (OR, 2.8; 95% CI, 1.2-5.5; P = 0.01). Cumulative exposure to DNAemia in the CMV blips greater than 910 IU/mL indicated increased HR of subsequent CMV infection (HR, 4.6; 95% CI, 1.2-17.2; P = 0.02). Cytomegalovirus blips are frequent; particularly when the viral load of the first positive PCR is < 910 IU/mL, and serostatus risk is intermediary/low. Accumulating blips suggest intermittent low-level replication. If blips are suspected, confirmation of ongoing replication before initiation of treatment is prudent.


Presystemic and systemic intestinal metabolism of fenoterol in the conscious rat.
The intestinal and liver first pass metabolism of fenoterol.HBr (Berotec, Partusisten) was investigated in the conscious rat. Fenoterol plasma concentrations (2-1000 ng/ml) were measured with a new HPLC determination using electrochemical detection. After intraduodenal administration, fenoterol was incompletely absorbed (47-66% not absorbed). Presystemic intestinal (EGpre) and liver (EH) extraction ratios, EGpre = 0.93 +/- 0.01, EH = 0.67 +/- 0.04, were calculated from AUC values after intraduodenal, intraportal, and iv administration. Saturation of intestinal and/or liver metabolism was checked by using three dose levels at different administration routes. Total systemic availability after intraduodenal administration ranged from 0.8 (10 mg/kg) to 1.2% (40 mg/kg). The contribution of the splanchnic region to the systemic clearance of fenoterol was assessed by measuring fenoterol and fenoterol-glucuronide concentrations in arterial and portal venous blood under steady state conditions. During iv infusion (30 micrograms fenoterol/min X kg), an intestinal extraction ratio of EG = 0.26 was observed. After iv administration of fenoterol (1 and 2 mg/kg), dose-dependent pharmacokinetics were observed. Doubling of the dose resulted in an increase of systemic clearance (Cl = 53.8 +/- 2.7 and 74.4 +/- 1.8 ml/min X kg) and distribution volume (Vss = 0.95 +/- 0.13 and 1.21 +/- 0.11 liters/kg); the mean residence time (17.9 +/- 2.4 and 16.3 +/- 1.4 min) and terminal half-life (45.8 +/- 5.5 and 46.8 +/- 2.8 min) were not changed.


The regulatory epicenter of miRNAs.
miRNAs are small non-coding RNAs with average length of ~21 bp. miRNA formation seems to be dependent upon multiple factors besides Drosha and Dicer, in a tissue/stage-specific manner, with interplay of several specific binding factors. In the present study, we have investigated transcription factor binding sites in and around the genomic sequences of precursor miRNAs and RNA-binding protein (RBP) sites in miRNA precursor sequences, analysed and tested in comprehensive manner. Here, we report that miRNA precursor regions are positionally enriched for binding of transcription factors as well as RBPs around the 3' end of mature miRNA region in 5' arm. The pattern and distribution of such regulatory sites appears to be a characteristic of precursor miRNA sequences when compared with non-miRNA sequences as negative dataset and tested statistically.When compared with 1 kb upstreamregions, a sudden sharp peak for binding sites arises in the enriched zone near the mature miRNA region. An expression-data-based correlation analysis was performed between such miRNAs and their corresponding transcription factors and RBPs for this region. Some specific groups of binding factors and associated miRNAs were identified. We also identified some of the overrepresented transcription factors and associated miRNAs with high expression correlation values which could be useful in cancer-related studies. The highly correlated groups were found to host experimentally validated composite regulatory modules, in which Lmo2-GATA1 appeared as the predominant one. For many of RBP-miRNAs associations, coexpression similarity was also evident among the associated miRNA common to given RBPs, supporting the Regulon model, suggesting a common role and common control of these miRNAs by the associated RBPs. Based on our findings, we propose that the observed characteristic distribution of regulatory sites in precursor miRNA sequence regions could be critical inmiRNA transcription, processing, stability and formation and are important for therapeutic studies. Our findings also support the recently proposed theory of self-sufficient mode of transcription by miRNAs, which states that miRNA transcription can be carried out in host-independent mode too.


Total knee arthroplasty with posterior cruciate retention in patients with rheumatoid arthritis.
The objective of the present study was to evaluate posterior cruciate ligament retention in total knee arthroplasty for patients with rheumatoid arthritis to determine long term ligamentuous stability. The study concerns an average 11-year followup (range, 10-13 years) of 52 patients (81 knees) with rheumatoid arthritis who had a total knee arthroplasty using a contemporary posterior cruciate retaining prosthesis. Particular attention was given to component survivorship and clinical stability. Fourteen patients (20 knees) died; none of these patients required revision surgery. No patients were lost to followup. Sixty-one knees in 38 patients were examined. In this group, the Knee Society knee score averaged 95 points (range, 63-100 points) and function score averaged 74 points (range, 0-100 points). Postoperative flexion averaged 112 degrees and extension averaged 0 degree. Four knees had 3 degrees asymptomatic hyperextension; one knee with 5 degrees hyperextension occasionally gave way. Five well aligned knees had between 6 degrees and 9 degrees varus or valgus laxity in extension, but no patient reported subjective instability. Two patients underwent revision surgery. One patient had a worn metal backed patella component replaced and the other patient had an open synovectomy for recurrent active rheumatoid synovitis. Thirteen-year survivorship based on need for revision surgery was 97% with the 95% confidence limits between 88% and 100%. There was no radiographic loosening or subsidence of prosthetic components. At 11-year followup, patients with rheumatoid arthritis whose knees were replaced with posterior cruciate retention prostheses experienced results equivalent to or better than those reported for patients with osteoarthritis at a similar followup. Late hyperextension and subsequent instability may be a concern in the second decade of followup.


A New Triage Method for Burn Disasters: Fast Triage in Burns (FTB).
BACKGROUND There are few triage methods based on patient age and extent of burn. However, in case of mass casualties, age might be hard to define. Burn segregation in mass-casualty accidents requires an easy, fast, and effective method. Triage in burns should also segregate casualties requiring treatment in burn centers. The aim of this study was to create a proprietary segregation algorithm dedicated to mass-casualty incidents. MATERIAL AND METHODS A retrospective analysis of 939 burned patients admitted to the Clinical Department of Burns, Plastic and Reconstructive Surgery, Military Institute of Medicine (MIM) in Warsaw and to the Center for the Treatment of Burns (CTB) in Siemianowice Śląskie in 2012 and 2013 was performed. The aim was to reveal which early factors could be used during segregation of burn victims in mass-casualty incidents on the battlefield and in civilian circumstances. Only easy and quick-to-evaluate factors that can be examined without medical equipment and laboratory tests were used in creating the proprietary triage algorithm. RESULTS As a result of our study, we created an algorithm for fast triage in mass-casualty situations. The algorithm is based on parameters that can be easily evaluated without additional equipment. To create the algorithm, we used factors that had the strongest impact on mortality prediction in severely burned patients, in multifactor analysis: advanced age (p<0.001. OR=1.04), extent of the deep burn (p<0.001. OR=1.1), and low systolic arterial pressure (p<0.001. OR=0.96). CONCLUSIONS The FTB (Fast Triage in Burns) algorithm is a new triage method dedicated for massive burn events in civilian circumstances. The FTB algorithm is a simple, quick, and credible means of segregating burn victims. The algorithm is dedicated to use in pre-hospital care, during mass-casualty events both in civilian and battlefield circumstances. The aim is to be able to evaluate burn victims immediately, without access to medical equipment or additional tests and to evaluate indications for burn center care. It is a unique method designed to be used during segregation in isolated burn mass-casualty incidents.


Synthesis of the 6,6,5,7-tetracyclic core of daphnilongeranin B.
An efficient approach toward the synthesis of the 6,6,5,7-tetracyclic core of the daphnilongeranin B, a Daphniphyllum alkaloid, is reported. The bridged 6,6-bicyclic system was constructed using a gold(i) catalysed Conia-ene reaction, while the 5- and 7-membered rings were assembled by two diastereoselective Michael addition reactions, respectively.


Post-traumatic endophthalmitis in children.
To study the clinico-microbiological profile and evaluate the functional and anatomical outcomes of pediatric post-traumatic endophthalmitis (PPTE). This was a retrospective interventional case series. All medical case records of patients diagnosed with PPTE over a 5-year period from January 2011 to December2015 were reviewed. Data recorded was: age, sex, type of trauma, mode of injury, interval between trauma and presentation, treatment, follow-up duration and final functional and anatomical outcomes. Forty-one eyes of 41 patients diagnosed with PPTE met the inclusion criteria. There were 26 boys and 15 girls. The mean age at the time of presentation was 7.34 years (1month -16years). The median interval between trauma and presentation was 3 days (mean = 13.71 days; 1-240 days). The average follow-up period was 292.24 days (median 150 days; 30 1440 days). Injury with wooden stick (20,50%) was the most common mode of injury. Culture positivity was noted in 25(61%) cases. Staphylococcusaureus was the most common organisms identified on culture. Univariate analysis of in dependent variables was done using the Chi-square test showed patients with positive culture for gram-positive organisms had better anatomical (p=0.038) and functional outcomes (p=0.043). 35(85%) patients underwent vitrectomy along with intraocular antibiotics. Optimal anatomical and functional outcomes were noted in 23(56.1%) and12(29.3%) respectively. PPTE carries a significantly poor prognosis in terms of ocular integrity and visual function. PPTE is common in boys and often caused by vegetative matter. Staphylococcus aureus is the most common organism seen in PPTE. Early presentation following trauma and identification of gram-positive organisms have a better prognosis.


Evaluation of intraosteoblastic activity of dalbavancin against Staphylococcus aureus in an ex vivo model of bone cell infection.
Long-acting lipoglycopeptides are promising therapeutic options in Staphylococcus aureus bone and joint infections (BJIs). This study evaluated the ability of dalbavancin to eradicate the intraosteoblastic reservoir of S. aureus, associated with BJI chronicity. Osteoblastic cells were infected with a standardized inoculum of the S. aureus reference strain HG001 and incubated for 24 h with dalbavancin, vancomycin or rifampicin using the MIC, 10×MIC, 100×MIC and/or the intraosseous concentrations reached using standard therapeutic doses (i.e. vancomycin, 10 mg/L; rifampicin, 2 mg/L; and dalbavancin, 6 mg/L). The remaining intracellular bacteria were quantified by plating cell lysates. MICs of dalbavancin, vancomycin and rifampicin were 0.125, 1 and 0.004 mg/L, respectively. Dalbavancin significantly reduced the intracellular inoculum of S. aureus starting at a concentration equal to the MIC, with a significant dose effect, ranging from a reduction of 31.4% (95% CI = 17.6%-45.2%) at MIC to 51.6% (95% CI = 39.8%-63.4%) at 100×MIC compared with untreated cells. Of note, dalbavancin was the only molecule to significantly reduce the intraosteoblastic inoculum at low concentration (MIC). At intraosseous concentrations, dalbavancin reduced the intracellular inoculum by 49.6% (95% CI = 45.1%-54.1%) compared with untreated cells (P < 0.001), with no significant difference compared with vancomycin (38.1%; 95% CI = 19.2%-57.0%; P = 0.646), and was less efficient than rifampicin (69.0%; 95% CI = 63.2-74.8; P < 0.001). Dalbavancin was able to decrease the intraosteoblastic S. aureus inoculum by 50% at intraosseous concentrations reached during standard human therapeutic dosing, with no difference compared with vancomycin, and remained less efficient than rifampicin. However, it was the only molecule significantly active at low concentration.


Trajectory study of supercollision relaxation in highly vibrationally excited pyrazine and CO2.
Classical trajectory calculations were performed to simulate state-resolved energy transfer experiments of highly vibrationally excited pyrazine (E(vib) = 37,900 cm(-1)) and CO(2), which were conducted using a high-resolution transient infrared absorption spectrometer. The goal here is to use classical trajectories to simulate the supercollision energy transfer pathway wherein large amounts of energy are transferred in single collisions in order to compare with experimental results. In the trajectory calculations, Newton's laws of motion are used for the molecular motion, isolated molecules are treated as collections of harmonic oscillators, and intermolecular potentials are formed by pairwise Lennard-Jones potentials. The calculations qualitatively reproduce the observed energy partitioning in the scattered CO(2) molecules and show that the relative partitioning between bath rotation and translation is dependent on the moment of inertia of the bath molecule. The simulations show that the low-frequency modes of the vibrationally excited pyrazine contribute most to the strong collisions. The majority of collisions lead to small DeltaE values and primarily involve single encounters between the energy donor and acceptor. The large DeltaE exchanges result from both single impulsive encounters and chattering collisions that involve multiple encounters.


The frequency and efficiency of endogene suppression by transitive silencing signals is influenced by the length of sequence homology.
Transitivity, the spread of RNA silencing along primary target sequences, leads to the degradation of secondary targets that have no sequence homology to the initial silencing trigger. We demonstrate that increasing the distance between direct and adjacent target sequences in a transgenic primary target delays the onset of silencing of a secondary target gene. Silencing can spread in a 3' to 5' direction over a distance of at least 500 nucleotides (nt), but this requires consistently more time compared to a distance of 98 nt or 250 nt. The efficiency and frequency of transitive silencing of an endogene depends on the length of its sequence homology with the primary target. With a length of 500 nt, efficient silencing can eventually be established in all plants, whereas lengths of 250 nt and 98 nt homology result in less efficient and less frequent suppression. These results suggest that amplification of secondary small interfering RNAs (siRNAs) is a time-requiring process that gradually expands the population of siRNAs until a steady-state level is reached. Moreover, the length of the sequence homology in the primary target providing secondary siRNAs determines whether this steady-state level readily exceeds the threshold necessary for efficient silencing.


Splenomegaly-associated thrombocytopenia after hepatic yttrium-90 radioembolization.
Thrombocytopenia is often observed after yttrium-90 radioembolization (RE). Possible mechanisms include radiation toxicity to the bone marrow, consumption in the liver due to local radiation effects, and sequestration in the spleen. We sought to identify the causative factors. Patients with complete baseline and 3-month post-RE imaging and laboratory data were included in this retrospective analysis. Univariate and multivariate regression analyses were performed on clinical, procedural, and imaging data. A total of 116 patients were identified (65 male, 51 female; median age 64 years). Forty-six patients were treated for primary and 70 for metastatic liver malignancy. Of these, 86 were treated with resin and 30 with glass microspheres; median activity was 1.85 GBq. Eighty-three patients underwent whole-liver treatment. Maximum individual change in platelet count was -20.2 % leading to new or increased grade of thrombocytopenia in 48 patients (41.4 %) by National Cancer Institute Common Toxicity Criteria for Adverse Events version 4.02 criteria. Independent risk factors for decreased platelet count included change in spleen volume (median change +17.5 %; p = 0.001) and whole-liver (rather than lobar or segmental) treatment (p = 0.025). Administered activity and absorbed dose were not associated with change in platelet count. The change in spleen volume itself was associated with previous epidermal growth factor receptor-inhibitor treatment (p = 0.002), whole-liver absorbed dose (p = 0.027), and multiple-line chemotherapy (0.012) for whole-liver treatments only. Post-RE treatment increase of spleen volume is correlated with decreased peripheral platelet count suggesting a mechanism of increased portal hypertension and hypersplenism being responsible.


Stereochemical analysis of leubethanol, an anti-TB-active serrulatane, from Leucophyllum frutescens.
Bioactivity-guided fractionation of the methanolic root bark extract of Leucophyllum frutescens led to the identification of leubethanol (1), a new serrulatane-type diterpene with activity against both multi-drug-resistant and drug-sensitive strains of virulent Mycobacterium tuberculosis. Leubethanol (1) was identified by 1D/2D NMR data, as a serrulatane closely related to erogorgiane (2), and exhibited anti-TB activity with minimum inhibitory concentrations in the range 6.25-12.50 μg/mL. Stereochemical evidence for 1 was gleaned from 1D and 2D NOE experiments, from 1H NMR full spin analysis, and by comparison of the experimental vibrational circular dichroism (VCD) spectrum to density functional theory calculated VCD spectra of two diastereomers.


Parathyroid hormone and parathyroid hormone-related peptide stimulate insulin-like growth factor-binding protein secretion by rat osteoblast-like cells through a adenosine 3',5'-monophosphate-dependent mechanism.
Specific insulin-like growth factor-binding proteins (IGFBPs) that may enhance or inhibit insulin-like growth factor (IGF) action are produced in various tissues. In the present study we demonstrated that IGFBPs are synthesized and secreted by rat osteoblast-like cells (UMR 106-01). PTH and PTH-related peptide (PTHrP) were potent stimuli for IGFBP production by UMR cells, whereas GH, IGF-I, insulin, epidermal growth factor, and T3 had little or no effect. A maximal 8- to 30-fold increase in IGFBP production was attained at 10(-7)-10(-6) M PTH and PTHrP, with a half-maximal effect at approximately 10(-9) M. By Western blot analysis, PTH and PTHrP markedly and selectively increased the production of 29,000 mol wt (Mr) and, to a lesser extent, 24,000 Mr IGFBPs. Agents that elevate intracellular cAMP by different mechanisms [(Bu)2cAMP, forskolin, and isobutylmethylxanthine] mimicked the effect of PTH and PTHrP on IGFBP synthesis. In comparison, PTH did not stimulate IGFBP production in fibroblasts and ROS 17/2.8 cells, which secrete IGFBPs of 42,000, 38,000, 34,000, 28,000, and 24,000 Mr, but not of 29,000 Mr. The PTH-responsive IGFBPs from UMR cells were nonglycosylated proteins with preferential affinity for IGF-I over IGF-II. These IGFBPs were not immunoprecipitated with antisera against rat IGFBP-2 or human IGFBP-1. Thus, PTH and PTHrP increase the production in UMR 106-01 cells of discrete IGFBP forms with Mr of 29,000 and 24,000 through a cAMP-mediated mechanism, independent of IGF-I synthesis. Taken with the known effects of PTH on IGF production in bone cells, the data suggest that PTH and PTHrP may modulate local IGF action in bone through the regulation of specific IGFBP availability.


A fermented soy permeate improves the skeletal muscle glucose level without restoring the glycogen content in streptozotocin-induced diabetic rats.
Exercise is essential into the therapeutic management of diabetic patients, but their level of exercise tolerance is lowered due to alterations of glucose metabolism. As soy isoflavones have been shown to improve glucose metabolism, this study aimed to assess the effects of a dietary supplement containing soy isoflavones and alpha-galactooligosaccharides on muscular glucose, glycogen synthase (GSase), and glycogen content in a type 1 diabetic animal model. The dietary supplement tested was a patented compound, Fermented Soy Permeate (FSP), developed by the French Company Sojasun Technologies. Forty male Wistar rats were randomly assigned to control or diabetic groups (streptozotocin, 45 mg/kg). Each group was then divided into placebo or FSP-supplemented groups. Both groups received by oral gavage, respectively, water or diluted FSP (0.1 g/day), daily for a period of 3 weeks. At the end of the protocol, glycemia was noticed after a 24-h fasting period. Glucose, total GSase, and the glycogen content were determined in the skeletal muscle (gastrocnemius). Diabetic animals showed a higher blood glucose concentration, but a lower glucose and glycogen muscle content than controls. Three weeks of FSP consumption allowed to restore the muscle glucose concentration, but failed to reduce glycemia and to normalize the glycogen content in diabetic rats. Furthermore, the glycogen content was increased in FSP-supplemented controls compared to placebo controls. Our results demonstrated that diabetic rats exhibited a depleted muscle glycogen content (-25%). FSP-supplementation normalized the muscle glucose level without restoring the glycogen content in diabetic rats. However, it succeeded to increase it in the control group (+20%).


Idiopathic Hypertrophic Spinal Pachymeningitis: A Diagnostic Challenge: A Case Report and Review of the Literature.
Idiopathic hypertrophic pachymeningitis (IHP) can resemble other disorders associated with spinal compression. It is a rare inflammatory fibrosing disease of the dura of unidentified etiology and is considered a diagnosis of exclusion. We present a case of idiopathic hypertrophic spinal pachymeningitis occupying a long segment of cervical dura. This is a case of 38-year-old female patient, who suffered progressive neck pain for 2-year duration. Examination revealed spasticity in all four limbs, plus three symmetric reflexes all over, and the sensory level at T4. Magnetic resonance imaging showed spinal cord compression by a thickened anterior and posterior dura adjacent to the cord from C2 to C7. The diagnosis of spinal IHP was confirmed through biopsy. The patient improved after treatment with corticosteroids. Early surgical intervention with postoperative corticosteroid therapy is a known treatment for this disease, as a way to prevent irreversible neurological damage.


[Density of the neurovegetative innervation of the uterus of rats].
Using the Karnovsky and Roots modified by E1 Badawi and Schenk's technic for the cholinergic innervation and with the method of Lindvall and Björklund modified by de la Torre and Surgeon for the adrenergic ones, we demonstrate the innervation of each tissular layer, in cervix, corpus and cornua of the uterus of the she-rat in pro-oestrus state. A statistic evaluation is established for the different areas. The density of cholinergic innervation is richer than the adrenergic one. The number of cholinergic fibers is maximum in the cervix. It decreases in corpus and cornua especially in the mucosal and sub-mucosal layers. The density of the muscular nervous network is predominant in the corpus versus the cervix. The number of adrenergic nerves is maximum in corpus it diminishes in the cervix and the cornua. In each region, the muscular adrenergic network domines over the other tissular layers. We confirm statistically morphometric non parametric observations of these other authors.


Tension free vaginal tape (TVT) vs Transobturator tape (TOT) complications and outcomes.
Stress urinary incontinence (SUI) is managed with both TVT and TOT. The TVT route of placement could result in a higher com-plication rate. The aim of this study is to analyze and compare outcomes and complications of TVT and TOT in SUI treatment. Prospective study in 64 women with isolated SUI was validated through Urogenital Distress Inventory (UDI-6), Incontinence Impact questionnaire (IIQ-7) and International Continence Impact Questionnaire (ICIQ5-SF). Intraoperative and postoperative complications were noted. Patients were considered cured when negative on stress test and with no need for additional surgery during follow up, after one and three months. The cure rates for both TVT 26/30 (86,6%) and TOT 30/34 (88,2) were comparable. Higher rates of bleeding, bladder perforations, pain and dyspareunia are recorded in TVT group. Quality of life graded on the questionnaire basis proved significant improvement in both procedures. Both TVT and TOT have comparable cure results in the treatment of SUI. The TOT is equally effective in the SUI treatment with significantly lower complication incidence.


Hemlock Alkaloids in Aloes. Occurrence and Distribution of gamma-Coniceine.
The hemlock alkaloid gamma-coniceine was identified in a number of ALOE species, namely A. GILLILANDII, Reynolds A. BALLYI Reynolds, A. RUSPOLIANA Baker, A. IBITIENSIS Perrier and A. DELTOIDEODONTA Baker. Coniine was identified in A. VIGUIERI Perrier. The levels of gamma-coniceine are higher than those found in CONIUM MACULATUM L. Some species also contained trace amounts of conhydrinone and pseudoconhydrin. Three of the species are Madagascan endemics, one is restricted to Arabia, while the rest are remote from each other in East Africa. Some of the species are loosely related but there is no overall taxonomic affinity between them.


Effect of cyclophosphamide or x-rays on spontaneously occurring metastases from tumors transplanted into the tails of mice.
A spontaneous metastases model in mice is being used to test the efficiency of various treatments in eliminating metastases. Solid tumors were transplanted into the tails of mice and removed by tail transection when they had grown to a 4- to 5- or 6- to 7-mm mean diameter. Subsequently, 70 to 95% of mice not given other treatment developed metastases in the lungs or in regional lymph nodes (lumbar sacral region), or in both sites. The present paper reports the effects of whole-body or partial-body treatment on these metastases. The treatments, which started at the time of surgical transection of the tail, included a range of single or fractionated doses of cyclophosphamide (CTX) or X-rays given either to the whole body or locally to the lungs only. CTX reduced the incidence of metastases in both sites although the incidence of lung metastases was reduced by smaller doses of CTX than that of the lumbar sacral metastases. Whole-body irradiation of 6 grays (600 rads) had no effect on the incidence of metastases, whereas local irradiation of the lungs with single doses of 14.5 or 20 grays reduced the number substantially, as did 95 mg or more of CTX per kg. Thus, CTX or radiation reduced the incidence of lung metastases in a system where metastases developed from cells seeded from a primary tumor rather than from a cell suspension injected into the tail vein.


Clinical evaluation of enteric media in the primary isolation of Salmonella and Shigella.
Over a 1-year period, media for the isolation of enteric pathogens were compared on 455 stool specimens. Fifty-three pathogens were isolated, of which 56% were Shigella sonnei and 13% were Sh. flexneri. Of these isolates, 90% were found on xylose-lysine-desoxycholate agar, 87% on Hekton enteric agar, and 80% on MacConkey without crystal violet with 2% agar and 0.007% neutral red, but only 28% were recovered on Salmonella-Shigella agar. Less than one-half of the shigellae were recovered after Selenite-F enrichment. On the other hand, enrichment was the most helpful method for isolating salmonellae. Studies on cultures from which mixed isolates were obtained indicated that numbers and chance distribution have an effect on the results obtained. The performance of Salmonella-Shigella agar in the isolation of enteric pathogens was inferior, and the effort involved to obtain those isolates was greater than for Hekton enteric and xylose-lysine-desoxycholate agars.


Prognostic significances of oxidative DNA damage evaluated by 8-hydroxy-deoxyguanosine and antioxidant enzymes in patients undergoing resection of gastric and colon carcinoma.
Oxidant/antioxidant balance has been suggested as an important factor for initiation and progression of cancer. In order to determine whether the degree of oxidative DNA damage and antioxidant enzyme activities in plasma obtained from patients with gastric and colon cancer who undergo resection can be used as a useful prognostic predictor, plasma level of 8-hydroxydeoxyguanosine (8-OHdG), activities of glutathione peroxidase (G-Px) and superoxide dismutase (SOD) were examined. 19 patients with gastric cancer and 26 patients with colon cancer who were undergoing resection of tumor were included by the study. Venous blood samples were taken just before the surgery. Plasma level of 8-OHdG was determined with ELISA, SOD and G-Px activities in plasma were measured by spectrophotometric kits. 8-OHdG level and activity of G-Px were found to be decreased, SOD activity was found to be increased in both gastric and colon cancer groups as compared to control group. Alpha fetoprotein was found to be correlated with G-Px in the gastric cancer group and correlated with 8-OHdG in the colon cancer group. SOD activity was correlated with CA-15-3 in the gastric cancer group. Low plasma level of 8-OHdG and altered antioxidant activity may implicate the deficient repair of oxidative DNA damage in patients with gastric and colon cancer. Those measured parameters were not found to be related with histopathological data but correlated with some tumor markers.


Sentinel lymph node evaluation does not improve staging accuracy in colon cancer.
Lymph node involvement is an important prognostic factor in colorectal cancer. Sentinel lymph node (SLN) evaluation for assessing lymph node status in colorectal cancer remains controversial. Here we evaluated the sensitivity, predictive value, and accuracy of SLN evaluation for determining lymph node status in resectable colon cancer. A prospective phase 2 cohort study of SLN evaluation in colon cancer was conducted from September 1998 to April 2006. Patients underwent resection and SLN mapping with 1% isosulfan blue and (m99)Tc sulfur colloid injection. SLNs were evaluated by hematoxylin and eosin (HE) staining and, if findings were negative, by additional thin HE sections and immunohistochemical (IHC) staining for pancytokeratin and MOC31. Overall survival for patients with IHC-positive disease was evaluated by Kaplan-Meier analysis and the log rank test. SLNs were identified in 119 (99%) of the 120 patients eligible for the study. Median number of SLNs identified was 4 (range, 0-13). Forty-nine patients (40%) had nodal metastases on HE. The SLN accurately identified nodal metastases in 29 (59%) of these 49 patients and was negative for metastases in 22 patients (41%). SLNs in eight patients (7%) were negative by HE but positive by IHC staining. Positive IHC status did not affect survival after a median follow-up of 33 months (P = .41). The low sensitivity and high false-negative rate of SLN evaluation does not support this technique for improving the accuracy of nodal staging for patients with colon cancer. The significance of IHC-positive SLNs remains uncertain.


Fluoride concentration of bottled water and public water in Lebanon.
People in Lebanon turned to bottle water consumption because of its poor public water quality. In general, fluoride is known as dental caries preventive. A study in 1986 and two other national surveys in 1994 and 2004 showed that the concentration of fluoride in public Lebanese water was insignificant for the prevention of dental caries (less than 0.3 mg/L). The aim of the study was to measure the fluoride concentration in the highest selling and known commercial brands for bottled water in Lebanon, as well as to assess their effectiveness regarding prevention of dental caries. Fluoride has a notable therapeutic effect but in small doses that fluoride can be found in drinking water. Analysis using an absorptiometry of 625nm and another technique using an Orion electrode of 9609 BN have shown the low content of fluoride in the Lebanese waters (less than 0.3 mg/l). Strategies have recently been evolved based on fluroide supplementation (if the results turn negative) to reduce the index of caries in LEBANON.


GATA2 regulates the erythropoietin receptor in t(12;21) ALL.
The t(12;21) (p13;q22) chromosomal translocation resulting in the ETV6/RUNX1 fusion gene is the most frequent structural cytogenetic abnormality in children with acute lymphoblastic leukemia (ALL). The erythropoietin receptor (EPOR), usually associated with erythroid progenitor cells, is highly expressed in ETV6/RUNX1 positive cases compared to other B-lineage ALL subtypes. Gene expression analysis of a microarray database and direct quantitative analysis of patient samples revealed strong correlation between EPOR and GATA2 expression in ALL, and higher expression of GATA2 in t(12;21) patients. The mechanism of EPOR regulation was mainly investigated using two B-ALL cell lines: REH, which harbor and express the ETV6/RUNX1 fusion gene; and NALM-6, which do not. Expression of EPOR was increased in REH cells compared to NALM-6 cells. Moreover, of the six GATA family members only GATA2 was differentially expressed with substantially higher levels present in REH cells. GATA2 was shown to bind to the EPOR 5'-UTR in REH, but did not bind in NALM-6 cells. Overexpression of GATA2 led to an increase in EPOR expression in REH cells only, indicating that GATA2 regulates EPOR but is dependent on the cellular context. Both EPOR and GATA2 are hypomethylated and associated with increased mRNA expression in REH compared to NALM-6 cells. Decitabine treatment effectively reduced methylation of CpG sites in the GATA2 promoter leading to increased GATA2 expression in both cell lines. Although Decitabine also reduced an already low level of methylation of the EPOR in NALM-6 cells there was no increase in EPOR expression. Furthermore, EPOR and GATA2 are regulated post-transcriptionally by miR-362 and miR-650, respectively. Overall our data show that EPOR expression in t(12;21) B-ALL cells, is regulated by GATA2 and is mediated through epigenetic, transcriptional and post-transcriptional mechanisms, contingent upon the genetic subtype of the disease.


Infection of CD34(+) hematopoietic progenitor cells by human herpesvirus 7 (HHV-7).
To investigate the tropism of the T-lymphotropic human herpesvirus 7 (HHV-7) for hematopoietic progenitors, cord blood CD34(+) cells were inoculated in vitro with HHV-7 and then induced to differentiate along the granulocytic and erythroid lineages by the addition of appropriate cytokine cocktails. In semisolid assays, HHV-7 modestly affected the growth of committed (granulocytic/macrophagic and erythroid) progenitors, whereas it significantly decreased the number of pluripotent (granulocytic/erythroid/ monocytic/megakaryocytic) progenitors. Such inhibitory effect was completely abrogated by incubating HHV-7 inoculum with anti-HHV-7 neutralizing serum. In liquid cultures, HHV-7 hastened maturation along the myeloid but not the erythroid lineage, as demonstrated by the up-regulation of CD33 early myeloid antigen at day 7 of culture, and of CD15 and CD14 antigens at day 15. Moreover, HHV-7 messenger RNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in cells maturating along both the myeloid and the erythroid lineages. To evaluate the relevance of these in vitro findings, the presence of HHV-7 was investigated in bone marrow (BM) unfractionated mononuclear cells (MCs) as well as in purified CD34(+) and CD34(-) cell subsets, obtained from 14 normal adult donors. HHV-7 DNA was detected by DNA-PCR in 4 of 7 BMMC samples, and it was found to be associated with both the CD34(-) (2 of 7) and the CD34(+ )(1 of 7) fractions. These data indicate that HHV-7 infects BM cells in vivo and shows the ability to affect the survival/differentiation of CD34(+) hematopoietic progenitors in vitro by inhibiting more ancestral progenitors and perturbing the maturation of myeloid cells.


Phosphorylation of hydroxylysine residues in collagen synthesized by cultured aortic smooth muscle cells.
O5-Phosphohydroxylysine was chemically synthesized and techniques were established for its identification by combined use of cation-exchange chromatography, thin-layer electrophoresis at pH 1.9 and 3.5, and thin-layer chromatography. Clean separation of phosphohydroxylysine from the other phospho amino acids, phosphoethanolamine, and phosphocholine was achieved. Conditions were also determined to permit hydrolysis of proteins in 2 M HCl without loss of the phosphono group of phosphohydroxylysine residues. Experiments were then performed showing that 32P was incorporated into the hydroxylysine residues of cell-associated collagens when cultured calf aorta medial smooth muscle cells were incubated with [32P]orthophosphate. In other experiments, the cells incorporated [3H]lysine into hydroxylysine residues of cell-associated collagen and then 32P into phosphohydroxylysine residues. The doubly labeled phosphohydroxylysine subsequently isolated showed nearly 1:1 stoichiometry with respect to incorporation of precursor lysine and phosphorus. Finally, in preliminary experiments done with a cell-free extract of the smooth muscle cells, 32P was transferred from [gamma-32P]ATP to hydroxylysine residues in several kinds of collagenous substrates. Thus, this work shows that smooth muscle cells have the capacity to phosphorylate hydroxylysine residues in their cell-associated collagens and provides preliminary evidence that a protein kinase is involved.


Topology- and size-dependent binding of DNA nanostructures to the DNase I.
The susceptibility of DNA nanomaterials to enzymatic degradation in biological environments is a significant obstacle limiting their broad applications in biomedicine. While DNA nanostructures exhibit some resistance to nuclease degradation, the underlying mechanism of this resistance remains elusive. In this study, the interaction of tetrahedral DNA nanostructures (TDNs) and double-stranded DNA (dsDNA) with DNase I is investigated using all-atom molecular dynamics simulations. Our results indicate that DNase I can effectively bind to all dsDNA molecules, and certain key residues strongly interact with the nucleic bases of DNA. However, the binding of DNase I to TDNs exhibits a non-monotonic behavior based on size; TDN15 and TDN26 interact weakly with DNase I (~ - 75 kcal/mol), whereas TDN21 forms a strong binding with DNase I (~ - 110 kcal/mol). Furthermore, the topological properties of the DNA nanostructures are analyzed, and an under-twisting (~32°) of the DNA helix is observed in TDN15 and TDN26. Importantly, this under-twisting results in an increased width of the minor groove in TDN15 and TDN26, which primarily explains their reduced binding affinity to DNase I comparing to the dsDNA. Overall, this study demonstrated a novel mechanism for local structural control of DNA at the nanoscale by adjusting the twisting induced by length.


Natural agents in the prevention of cancer, part two: preclinical data and chemoprevention for common cancers.
This paper is the second of a series examining the use of nutritional supplements as chemopreventive agents. The animal and in vitro data are reviewed in support of their use. Human safety data and mechanisms of action are described as well. Many over-the-counter dietary supplements have been shown to have significant chemopreventive activity in preclinical studies. Few side effects are associated with even long-term use of these agents. Along with dietary and lifestyle risk-reducing strategies, nutritional supplementation appears to be a viable intervention for those considered to be at high risk of developing cancer.


Effects of music therapy under general anesthesia in patients undergoing abdominal surgery.
Music therapy, an innovative approach that has proven effectiveness in many medical conditions, seems beneficial also in managing surgical patients. The aim of this study is to evaluate its effects, under general anesthesia, on perioperative patient satisfaction, stress, pain, and awareness. This is a prospective, randomized, double-blind study conducted in the operating theatre of visceral surgery at Sahloul Teaching Hospital over a period of 4 months. Patients aged more than 18 undergoing a scheduled surgery under general anesthesia were included. Patients undergoing urgent surgery or presenting hearing or cognitive disorders were excluded. Before induction, patients wore headphones linked to an MP3 player. They were randomly allocated into 2 groups: Group M (with music during surgery) and group C (without music). Hemodynamic parameters, quality of arousal, pain experienced, patient's satisfaction, and awareness incidence during anesthesia were recorded. One hundred and forty patients were included and allocated into 2 groups that were comparable in demographic characteristics, surgical intervention type and anesthesia duration. Comparison of these two groups regarding the hemodynamic profile found more stability in group M for systolic arterial blood pressure. A calm recovery was more often noted in group M (77.1% versus 44%, p < 10-3). The average Visual Analog Scale (VAS) score was lower in the intervention group (33.8 ± 13.63 versus 45.1 ± 16.2; p < 10-3). The satisfaction rate was significantly higher among the experimental group (81.4% versus 51.4%; p < 10-3). The incidence of intraoperative awareness was higher in group C (8 cases versus 3 cases) but the difference was not statistically significant. Music therapy is a non-pharmacological, inexpensive, and non-invasive technique that can significantly enhance patient satisfaction and decrease patients' embarrassing experiences related to perioperative stress, pain, and awareness.


Improving the prioritization of children at the emergency department: Updating the Manchester Triage System using vital signs.
Vital signs are used in emergency care settings in the first assessment of children to identify those that need immediate attention. We aimed to develop and validate vital sign based Manchester Triage System (MTS) discriminators to improve triage of children at the emergency department. The TrIAGE project is a prospective observational study based on electronic health record data from five European EDs (Netherlands (n = 2), United Kingdom, Austria, and Portugal). In the current study, we included 117,438 consecutive children <16 years presenting to the ED during the study period (2012-2015). We derived new discriminators based on heart rate, respiratory rate, and/or capillary refill time for specific subgroups of MTS flowcharts. Moreover, we determined the optimal cut-off value for each vital sign. The main outcome measure was a previously developed 3-category reference standard (high, intermediate, low urgency) for the required urgency of care, based on mortality at the ED, immediate lifesaving interventions, disposition and resource use. We determined six new discriminators for children <1 year and ≥1 year: "Very abnormal respiratory rate", "Abnormal heart rate", and "Abnormal respiratory rate", with optimal cut-offs, and specific subgroups of flowcharts. Application of the modified MTS reclassified 744 patients (2.5%). Sensitivity increased from 0.66 (95%CI 0.60-0.72) to 0.71 (0.66-0.75) for high urgency patients and from 0.67 (0.54-0.76) to 0.70 (0.58-0.80) for high and intermediate urgency patients. Specificity decreased from 0.90 (0.86-0.93) to 0.89 (0.85-0.92) for high and 0.66 (0.52-0.78) to 0.63 (0.50-0.75) for high and intermediate urgency patients. These differences were statistically significant. Overall performance improved (R2 0.199 versus 0.204). Six new discriminators based on vital signs lead to a small but relevant increase in performance and should be implemented in the MTS.


Sulphate groups are involved in the antigenicity of keratan sulphate and mask i antigen expression on their poly-N-acetyllactosamine backbones. An immunochemical and chromatographic study of keratan sulphate oligosaccharides after desulphation or nitrosation.
Conditions were established for desulphation of hexa-, octa-, deca- and larger oligosaccharides derived from corneal keratan sulphate after treatment with endo-beta-galactosidase. The antigenicities of the desulphated oligosaccharides were compared with those of the native oligosaccharides in chromatogram binding, plastic-plate binding or inhibition of binding assays using a novel microimmunochemical approach with oligosaccharide-lipid conjugates (neoglycolipids). The results clearly show that sulphate residues are essential components of the antigenic determinant(s) recognised by three monoclonal antibodies to keratan sulphate, 5-D-4, 1-B-4 and MZ15, but they mask the i antigen activity of the linear poly-(N-acetyllactosamine) backbones of this glycosaminoglycan. Immunochemical assays, before and after beta-N-acetylglucosaminidase treatment of desulphated linear hexa-, octa- and decasaccharides derived from keratan sulphate, indicate that for reaction with one anti-i antibody, Den, there is an absolute requirement for the non-reducing beta-galactosyl residue of the i antigen structure to be in the terminal position, but with a second anti-i antibody, Tho, there is in addition some reactivity with the i antigen structure having an N-acetylglucosamine residue at the non-reducing end. The chromatographic properties after desulphation or nitrosation of a minor keratan sulphate oligosaccharide (a dodecasaccharide), which reacts especially well with antibody 5-D-4, have provided the first evidence for the presence of glucosamine residues that may be N-sulphated in corneal keratan sulphate.


Preferences for long-term care services: willingness to pay estimates derived from a discrete choice experiment.
Ageing populations increase pressure on long-term care. Optimal resource allocation requires an optimal mix of care services based on costs and benefits. Contrary to costs, benefits remain largely unknown. This study elicits preferences in the general elderly population for long-term care services for varying types of patients. A discrete choice experiment was conducted in a general population subsample aged 50-65 years (N = 1082) drawn from the Dutch Survey Sampling International panel. To ascertain relative preferences for long-term care and willingness to pay for these, participants were asked to choose the best of two care scenarios for four groups of hypothetical patients: frail and demented elderly, with and without partner. The scenarios described long-term care using ten attributes based on Social Production Function theory: hours of care, organized social activities, transportation, living situation, same person delivering care, room for individual preferences, coordination of services, punctuality, time on waiting list, and co-payments. We found the greatest value was attached to same person delivering care and transportation services. Low value was attached to punctuality and room for individual preferences. Nursing homes were generally considered to be detrimental for well-being except for dementia patients without a partner. Overall, long-term care services were thought to produce greatest well-being for the patients 'without a partner' and those 'with dementia'. Individuals combining these two risk factors would benefit the most from all services except transportation which was considered more important for the frail elderly. The results support the notion that long-term care services represent different value for different types of patients and that the value of a service depends upon the social context. Examination of patient profiles confirmed the notion that physical, mental and social vulnerability affect valuation of the services. Policy-making would profit from allocation models in which budgetary requirements of different services can be balanced against the well-being they produce for individuals.


Maternal and developmental toxicity evaluation of melatonin administered orally to pregnant Sprague-Dawley rats.
Melatonin (MEL) is a widely used, over-the-counter sleep aid, and it has putative contraceptive, antioxidant, antiaging, and anticancer effects. The developmental toxicity potential for repeated oral doses of MEL had not previously been evaluated. In the present studies, time-mated, Sprague-Dawley-derived (CD) rats were administered MEL or vehicle by gavage on gestation days (gd) 6-19. MEL-treated groups received 1-, 10-, 100-, 150-, or 200-mg/kg body weight/day in the screening study (15 rats/group), and 50, 100, or 200 mg/kg/day in the definitive study (25 rats/group). In both studies, maternal food/water consumption, body weight, and clinical signs were monitored at regular intervals throughout gestation. At termination (gd 20, both studies), maternal liver and gravid uterine weights, number of ovarian corpora lutea, conceptus survival, fetal sex, and fetal body weight were evaluated. Fetal morphological examination included external structures (both studies) as well as visceral and skeletal structures (definitive study). In the screening study, maternal serum levels of 17beta-estradiol, progesterone, prolactin, and luteinizing hormone were determined by radioimmunoassay, and mammary tissue was fixed, stained, and evaluated for percent glandular area within the fat pad. No maternal morbidity/mortality was found in either study. In the screening study, aversion to treatment (> or =100 mg/kg/day) and reduced maternal weight gain (> or =150 mg/kg/day) were noted, but reproductive/endocrine parameters and fetal development were not affected. In the definitive study, aversion to treatment was noted at > or =50 mg/kg/day, and mild sedation, reduced maternal food intake, and reduced body weight gain were found during initial treatment with 200 mg/kg/day. MEL had no effect on prenatal survival, fetal body weight, or incidences of fetal malformations/variations. Thus, in the definitive study, the maternal toxicity NOAEL and LOAEL were 100 and 200 mg/kg/day, respectively, and the developmental toxicity NOAEL was > or =200 mg/kg/day.


Role of the Long Non-Coding RNA Growth Arrest-Specific 5 in Glucocorticoid Response in Children with Inflammatory Bowel Disease.
Glucocorticoids (GCs) are widely employed in inflammatory, autoimmune and neoplastic diseases, and, despite the introduction of novel therapies, remain the first-line treatment for inducing remission in inflammatory bowel disease (IBD). Given the high incidence of suboptimal response, associated with a significant number of side-effects, that are particularly severe in paediatric patients, the identification of subjects that are most likely to respond poorly to GCs is extremely important. Recent evidence suggests that the long non-coding RNA (lncRNA) GAS5 could be a potential marker of GC resistance. To address this issue, we evaluated the association between the lncRNA GAS5 and the efficacy of steroids, in terms of inhibition of proliferation, in two cell lines derived from colon and ovarian cancers, to confirm the sensitivity and specificity of these lncRNAs. These cells showed a different sensitivity to GCs and revealed differential expression of GAS5 after treatment. GAS5 was up-regulated in GC-resistant cells and accumulated more in the cytoplasm compared to the nucleus in response to the drug. The functions of GAS5 were assessed by silencing, and we found that GAS5 knock-down reduced the proliferation during GC treatment. Furthermore, for the first time, we measured GAS5 levels in 19 paediatric IBD patients at diagnosis and after the first cycle of GCs, and we demonstrated an up-regulation of the lncRNA in patients with unfavourable steroid response. Our preliminary results indicate that GAS5 could be considered a novel pharmacogenomic marker useful for the personalization of GC therapy.


A rapid method for the purification of glucose-6-phosphate dehydrogenase from bovine lens.
This paper describes a simple and rapid method for the purification of glucose-6-phosphate dehydrogenase from bovine lens, together with analysis of the kinetic behaviour and some properties of the enzyme. The purification consisted of two steps, 2',5'-ADP-Sepharose 4B affinity chromatography and DEAE Sepharose Fast Flow ion exchange chromatography in procedure which took two working days. The enzyme was obtained with a yield of 13.7% and had a specific activity of 2.64 U/mg protein. The overall purification was about 19,700-fold. The molecular weight of the enzyme was found to be 62 +/- 3 kDa by Sephadex G-200 gel filtration chromatography. A protein band corresponding to a molecular weight of 69.2 +/- 3.2 kDa was obtained on SDS polyacrylamide slab gel electrophoresis. On chromatofocusing, lens glucose-6-phosphate dehydrogenase gave a single peak at pI 5.14. The activation energy of the reaction catalyzed by the enzyme was calculated from Arrhenius plot as Ea = 5.88 kcal/mol. The pH versus velocity curve had two peaks at pH 7.7 and 9.6. By the double-reciprocal plots and the product inhibition studies, it was shown that the enzyme follows 'Ordered Bi Bi' sequential kinetics. From the graphical and statistical analyses, KmNADP+, KmG-6-P, KiNADPH, Ki6-PGA were estimated to be 0.008 +/- 0.002, 0.035 +/- 0.013, 0.173 +/- 0.007 and 1.771 +/- 0.160 mM, respectively. The observed kinetic behaviour of glucose-6-phosphate dehydrogenase from bovine lens was in accordance with the enzyme from other sources.


Maternal Hypertensive Disorders in Pregnancy and Postpartum Plasma B Vitamin and Homocysteine Profiles in a High-Risk Multiethnic U.S., Population.
Background: Hypertensive disorders of pregnancy are a recognized risk factor of a woman's future cardiovascular risk. The potential role of micronutrients in mitigating hypertensive disorders is not fully understood. This study examined maternal postpartum plasma B vitamin profiles by hypertensive disorders of pregnancy in a high-risk multiethnic U.S. population. Materials and Methods: The analyses included 2584 mothers enrolled within 3 days postpartum at the Boston Medical Center. Hypertensive disorders of pregnancy included gestational hypertension and pre-eclampsia disorders (pre-eclampsia, eclampsia, hemolysis, elevated liver enzymes, and/or low platelets syndrome) as documented in the medical records. Plasma folate, vitamin B12, and homocysteine levels were measured in blood samples collected at enrollment. Kernel density plots and multivariable regressions were used to examine the relationship between hypertensive disorders and postpartum B vitamin profiles. Results: Of the 2584 mothers, 10% had pre-eclampsia disorders that were associated with significantly lower plasma folate (adjusted beta coefficient (aβ): -0.10; 95% CI: -0.22 to -0.06) and increased homocysteine (aβ: 0.08; 95% CI: 0.04-0.13), but not with vitamin B12 concentrations. These associations remained robust after adjusting for a range of pertinent covariables and were more pronounced in non-Hispanic Black women compared with other groups. However, gestational hypertension was not significantly associated with any postpartum biomarker. Conclusions: We found that pre-eclampsia disorders, but not gestational hypertension, was associated with lower folate and higher homocysteine levels postpartum, especially among Black mothers. This finding, if further confirmed, may have implications for postpartum care, including attention to maternal micronutrient status to reduce and prevent hypertensive disorders in pregnancy-associated consequences in subsequent pregnancies and lifespan. Registration date: July 25, 2017; Registry website: https://clinicaltrials.gov/ct2/show/NCT03228875.


A multicenter, randomized comparison between 2, 5, and 8 mg of perineural dexamethasone for ultrasound-guided infraclavicular block.
This multicenter, randomized trial compared 2, 5, and 8 mg of perineural dexamethasone for ultrasound-guided infraclavicular brachial plexus block. Our research hypothesis was that all three doses of dexamethasone would result in equivalent durations of motor block (equivalence margin=3.0 hours). Three hundred and sixty patients undergoing upper limb surgery with ultrasound-guided infraclavicular block were randomly allocated to receive 2, 5, or 8 mg of preservative-free perineural dexamethasone. The local anesthetic agent (35 mL of lidocaine 1%-bupivacaine 0.25% with epinephrine 5 µg/mL) was identical in all subjects. Patients and operators were blinded to the dose of dexamethasone. During the performance of the block, the performance time, number of needle passes, procedural pain, and complications (vascular puncture, paresthesia) were recorded. Subsequently a blinded observer assessed the success rate (defined as a minimal sensorimotor composite score of 14 out of 16 points at 30 min), onset time as well as the incidence of surgical anesthesia (defined as the ability to complete surgery without local infiltration, supplemental blocks, intravenous opioids, or general anesthesia). Postoperatively, the blinded observer contacted patients with successful blocks to inquire about the duration of motor block, sensory block, and postoperative analgesia. The main outcome variable was the duration of motor block. No intergroup differences were observed in terms of technical execution (performance time/number of needle passes/procedural pain complications), onset time, success rate, and surgical anesthesia. Furthermore, all three doses of dexamethasone provided similar durations of motor block (14.9-16.1 hours) and sensory block. Although 5 mg provided a longer analgesic duration than 2 mg, the difference (2.7 hours) fell within our pre-established equivalence margin (3.0 hours). 2, 5, and 8 mg of dexamethasone provide clinically equivalent sensorimotor and analgesic durations for ultrasound-guided infraclavicular block. Further trials are required to compare low (ie, 2 mg) and ultra-low (eg, 0.5-1 mg) doses of perineural dexamethasone for brachial plexus blocks. TCTR20150624001.


Individual and combined effects of copper and parasitism on osmoregulation in the European eel Anguilla anguilla.
The European eel (Anguilla anguilla), a catadromous species, breeds in the sea and migrates to estuarine, lagoon or freshwater habitats for growth and development. Yellow eels, exposed to low or fluctuating salinities, are also exposed to multiple other stressors as pollution, over-fishing and parasitism, which contribute to the dramatic decrease of eel populations in several European countries. The objective of this study was to evaluate the single and combined effects of waterborne copper and experimental infestation of eels with the nematode Anguillicoloides crassus after a salinity challenge from nearly isotonic (18ppt) to hypo- (5ppt) and hypertonic (29ppt) conditions, in order to investigate the osmoregulatory capacity of eels exposed to these stressors. In a nearly isotonic condition (18ppt), blood osmolality remained constant over the 6 weeks contamination to Cu(2+) and Anguillicoloides crassus. In fish exposed to a salinity challenge of 29ppt for 2 weeks, no significant effect was recorded in blood osmolality, Na(+)/K(+)-ATPase (NKA) activity, Na(+) and Cl(-) concentrations. After 2 weeks at 5ppt however, a significant blood osmolality decrease was detected in fish exposed to Anguillicoloides crassus infestation with or without Cu(2+) addition. This decrease may originate from lower Cl(-) levels measured in eels exposed to both stressors. Blood Na(+) levels remained relatively stable in all tested animals, but gill NKA activities were lower in eels exposed to combined stress. No apparent branchial lesions were detected following the different treatments and immunolocalization of NKA revealed well-differentiated ionocytes. Thus, the 5ppt challenge in eels exposed to copper and Anguillicoloides crassus seems to clearly enhance iono/osmoregulatory disturbances. Funded by ANR CES/CIEL 2008-12.


[Accumulation of aflatoxins on wheat grain inoculated by Aspergillus flavus NRRL 2999 and their distribution in grinding products].
The influence of temperature ranging from 15 to 35 degrees C and relative humidity of 75 to 90% on the accumulation of aflatoxins on wheat grain inoculated by Aspergillus flavus NRRL 2999 was investigated. The lowest accumulation of aflatoxins took place at 20 degrees C and relative humidity of 80%. The highest accumulation of aflatoxins appeared in the grain after 3 day storage. Some products of three-graded 78% grinding showed a decrease in the aflatoxin content (flour of the first and second grade). During baking of leavened bread 70-80% of aflatoxins were decomposed. The feasible use of the rapid conditioning and Remix-method to reduce the content of aflatoxins in flour and bread is discussed.


Functional characterization of late embryogenesis abundant genes and promoters in pearl millet (Pennisetum glaucum L.) for abiotic stress tolerance.
Late embryogenesis abundant (LEA) genes display distinct functions in response to abiotic stresses in plants. In pearl millet (Pennisetum glaucum L.), a total of 21 PgLEA genes were identified and classified into six groups including LEA1, LEA2, LEA3, LEA5, LEA7, and dehydrins (DHN). Open reading frames (ORFs) of PgLEAs range from 291 bp (PgLEA1-1) to 945 bp (PgLEA2-11) and distributed randomly among the seven chromosomes. Phylogenetic analysis revealed that all PgLEA proteins are closely related to sorghum LEA proteins. The PgLEAs were found to be expressed differentially under high progressive vapor pressure deficit (VPD), PgLEA7 was significantly expressed under high VPD and was selected for functional validation. In silico analysis of the PgLEA promoter regions revealed abiotic stress-specific cis-acting elements such as ABRE, CCAAT, MYBS, and LTRE. Based on the type of motifs, PgLEAPC promoter (758 bp), its deletion 1 (PgLpd1, 349 bp) and deletion 2 (PgLpd2, 125 bp) were cloned into the plant expression vector pMDC164 having the promoter-less uidA gene. All the three plant expression vectors were introduced into tobacco through Agrobacterium tumefaciens-mediated transformation to obtain T1 and T2 generations of transgenic plants. Based on expression of the uidA gene, tissue-specific expression was observed in mature stems, roots and seedlings of PgLEAPC and PgLpd1 carrying transgenics only. While the transgenic PgLEAPC plants displayed significantly higher uidA expression in the stem and root tissues under salt, drought, heat, and cold stresses, very low or no expression was observed in PgLpd1 and PgLpd2 transgenics under the tested stress conditions. The results of this study indicate that the complete promoter of PgLEAPC plays a role in developing abiotic stress tolerance in plants.


C3 Targeted Complement Therapy for Chronic Periodontitis - A Scoping Review.
Chronic Periodontitis (CP) is a complex disease initiated by inflammation caused by dysbiotic bacterial communities in the subgingival environment. The Porphyromonas gingivalis, a keystone pathogen at low colonization, causes immune subversion of complement component C5aR, leading to complement C3-dependent destructive inflammation responsible for the inflammatory bone loss in CP. Animal studies have shown that targeting complement C3 with its inhibitor like AMY-101 may help reduce inflammatory bone loss in CP. This scoping review elaborates on the role of complement C3 targeted therapy for CP. About 66 original studies were obtained during an initial electronic search in Medline (Pubmed), Scopus, Web of Science, and Embase. About four articles were included in the review after screening the duplicates and reading the full text. Their aims and objectives, drug dosage, route of administration, results, and conclusions were recorded. Of the four-original research, 3 were animal studies and one randomized Phase IIa clinical trial. They showed that C3 targeted complement therapy reduced the inflammatory and clinical periodontal parameters in CP. C3 targeted complement therapy may be regarded as a valuable adjunct to non-surgical periodontal treatment for CP. However, the results are still under investigation and require further verification through clinical trials.


GAPs in Slit-Robo signaling.
Neuronal migration requires the integration of a number of diverse environmental cues and subsequent translation to specific responses such as directed cytoskeletal remodeling. Accurate knowledge of the signal transduction pathways linking activation of surface receptors to actin dynamics is necessary in order to understand the regulation of such processes. Activation of the Roundabout (Robo) receptor mediates a repulsive response in certain pioneering axons and migratory neurons. Recently, Wong et al have described a signaling link between Robo activation and specific GTPase activity that appears to regulate neuronal migration. A novel family of GTPase regulators, responsive to Slit-Robo engagement, has been identified and convincingly shown to alter the migration of neuronal cells. This study not only delineates a specific signaling route from guidance receptors to directed neuronal movement, but also offers clues towards potential regulatory mechanisms that ensure specificity of the Slit-Robo response.


Dietary sodium and iodine in remote Indigenous Australian communities: will salt-reduction strategies increase risk of iodine deficiency? A cross-sectional analysis and simulation study.
Excess salt intake is a global issue. Effective salt-reduction strategies are needed, however, as salt is a vehicle for iodine fortification, these strategies may also reduce iodine intake. This study examines the case of the remote Indigenous Australian population; we employed an innovative, objective method to assess sodium and iodine intakes against requirements and modelled the potential effects of salt-reduction strategies on estimated sodium and iodine intakes. Store-sales data were collected from 20 remote Indigenous community stores in 2012-14 representing the main source of food for 2 years for ~8300 individuals. Estimated average sodium and iodine intakes were compared against recommendations (nutrient reference values weighted to age and gender distribution). Linear programming was employed to simulate potential effects of salt-reduction strategies on estimated sodium and iodine intakes. Estimated average sodium intake was 2770 (range within communities 2410-3450) mg/day, far exceeding the population-weighted upper limit (2060 mg/day). Discretionary (added) salt, bread and processed meat were the biggest contributors providing 46% of all sodium. Estimated average iodine intake was within recommendations at 206 (186-246) μg/day. The following scenarios enabled modelling of estimated average salt intake to within recommendations: 1) 67% reduction in sodium content of bread and discretionary salt intake, 2) 38% reduction in sodium content of all processed foods, 3) 30% reduction in sodium content of all processed foods and discretionary salt intake. In all scenarios, simulated average iodine intakes remained within recommendations. Salt intakes of the remote Indigenous Australian population are far above recommendations, likely contributing to the high prevalence of hypertension and cardiovascular mortality experienced by this population. Salt-reduction strategies could considerably reduce salt intake in this population without increasing risk of iodine deficiency at the population-level. These data add to the global evidence informing salt-reduction strategies and the evidence that these strategies can be synergistically implemented with iodine deficiency elimination programmes. Australian New Zealand Clinical Trials Registry: ACTRN12613000694718.


Co-infection of disseminated histoplasmosis and tuberculosis in an AIDS patient.
Histoplasmosis is a fungal disease caused by the dimorphic fungus Histoplasma capsulatum, recognized as an AIDS-defining illness since the Center for Disease Control's revision criteria in 1985. This infection is reported to be present in 5-20% of AIDS patients, and in 95% of the cases it is manifested in its disseminated form. Serum antibodies and/or antigen research can make diagnosis, but the demonstration of the agent by culture or histopathological examination remains the gold standard methods. Co-infections in patients with AIDS are well known; however, reports on disseminated tuberculosis and histoplasmosis are scarce. The authors report the case of a female patient who presented a short-course history of weight loss, fever, and mild respiratory symptoms, with hepatosplenomegaly and lymphadenopathy. Laboratory workup called attention to anemia, altered liver, canalicular enzymes, liver function tests, high titer of lactate dehydrogenase (LDH), and pulmonary nodules on thoracic computed tomography. Incidental finding of yeast forms within the leukocytes during a routine blood cell count highlighted the diagnosis of histoplasmosis. The patient started receiving amphotericin B but succumbed soon after. The authors emphasize the possibility of this co-infection, the diagnosis of severe infection through the finding of yeast forms within peripheral leukocytes, and for the high titer of LDH in aiding the differential diagnosis.


Within-canopy sampling of global irradiance to describe downwelling light distribution and infer canopy stratification in a broadleaf forest.
A broadleaf mixed forest diversified through partial tree thinning was studied to identify expedient sampling and data analysis procedures to capture the heterogeneous within-canopy downward distribution of instantaneous global photosynthetic photon flux (PPF); to extract foliage structural properties from the acquired light values; and to compute statistics descriptive of the within-canopy light and leaf layer distributions. We sampled PPF at 1-m intervals along vertical gradients using a helium-filled balloon as a platform for a light sensor. A random method was used to identify the forest floor locations for the within-canopy balloon ascents. About 400 PPF measurements were recorded per vertical transect. For each PPF value, we computed, by inversion of the Monsi-Saeki model, the number of leaf strata cumulated along the sunbeam direction from the position where the light was measured. Variability in PPF and leaf layer at different vegetation scales was computed by non-parametric statistics. The methods were evaluated as appropriate for intra-canopy PPF sampling, particularly in an undisturbed canopy. The minimum number of vertical PPF profiles required to capture the within-canopy PPF variability was 9-10 (equivalent to about 4000 measurements). The reliability and sensitivity of the inversion of the Monsi-Saeki method were sufficient to capture the canopy structural differences between undisturbed and partially thinned forests. The proposed PPF canopy sampling and data analysis procedures provide a fast, reliable and inexpensive way to characterize tree crown structure, and to predict plant growth and forest dynamics and could be applied whenever vegetation absorbed radiation is a main driving force for forest canopy processes. The experimental light attenuation data and the extracted canopy leaf layer numbers could serve to corroborate canopy mechanistic models of radiative transfer and net primary production.