Identification of different critical embryonic periods to modify egg incubation temperature in mule ducks.
Egg incubation of mule ducks, mainly used for fatty liver production, is one of the critical phases in this sector. Based on hatching rate, the best incubation parameters have already been well described for poultry, but the literature on ducks is lacking. In this study, we tested different incubation conditions by varying two important factors, temperature and relative humidity, in mule ducks. These variations were applied at different periods during embryogenesis in order to measure the impact of environmental disturbances on different zootechnical performances. The temperature was increased by 1.5 °C (16 h/24) and the relative humidity was set up to 65%, during 10 days. Six 10-day developmental windows were tested, from embryonic day 9 to embryonic day 14. Our results are in line with previous reports showing that increasing incubation temperature, even when relative humidity is adjusted, can have a negative impact on duck embryonic mortality up to 24.5% for the condition E10-E20 (P < 10-5). However, the hatchability can be maintained at the level of the control groups when these modifications are applied on the latest windows (from the 11th embryonic day). Sex ratio, hatching BW, and internal temperature are also sensitive to these incubation changes, and their modification could have a major impact on later zootechnical performance. These results should contribute to the development or embryonic temperature programming approaches, especially for the fatty liver production industry.


Developmental Changes in Dendritic Spine Morphology in the Striatum and Their Alteration in an A53T α-Synuclein Transgenic Mouse Model of Parkinson's Disease.
The aging process is accompanied by various neurophysiological changes, and the severity of neurodegenerative disorders such as Parkinson's disease (PD) increases with aging. However, the precise neuroanatomical changes that accompany the aging process in both normal and pathologic conditions remain unknown. This is in part because there is a lack of high-resolution imaging tool that has the capacity to image a desired volume of neurons in a high-throughput and automated manner. In the present study, focused ion beam/scanning electron microscopy (FIB/SEM) was used to image striatal neuropil in both wild-type (WT) mice and an A53T bacterial artificial chromosome (BAC) human α-synuclein (A53T-BAC-SNCA) transgenic (Tg) mouse model of PD, at 1, 3, 6, and 22 months of age. We demonstrated that spine density gradually decreases, and average spine head volume gradually increases with age in WT mice, suggesting a homeostatic balance between spine head volume and spine density. However, this inverse relationship between spine head volume and spine density was not observed in A53T-BAC-SNCA Tg mice. Taken together, our data suggest that PD is accompanied by an abnormality in the mechanisms that control synapse growth and maturity.


Role of Janus Kinase (JAK) Inhibitor in Autoimmune Ocular Inflammation: A Systematic Review.
To evaluate the effectiveness of Janus kinase (JAK) inhibitors for the treatment of patients with autoimmune disease and associated inflammatory ocular diseases. We identified relevant literature by screening the MEDLINE, PubMed, and Cochrane databases for randomized controlled trials, cohort studies, case controls, and case reports. Seven studies, including 11 patients, were included in the final systematic analysis. Of the 11 patients, there were 5 cases of juvenile idiopathic arthritis- (JIA-) associated uveitis, 1 case of rheumatoid arthritis- (RA-) associated keratitis, 1 case of RA-associated scleritis, 1 case of psoriasis-associated conjunctivitis, 2 cases of noninfectious scleritis, and 1 case of uveitis with suspected autoimmune disease. None of these 11 patients responded adequately to conventional treatments, including biological agents; these were all refractory cases and switched to JAK inhibitor therapy. Irrespective of whether they were suffering from uveitis, scleritis, or other types of ocular inflammation, all 11 patients showed an improvement to JAK inhibitors without significant side effects. Different types of JAK inhibitors might be associated with different responses when used to treat ocular inflammation. JAK inhibitors may represent an alternative treatment option for patients with autoimmune ocular inflammation.


Rapture of renal angiomyolipoma during pregnancy: a case report.
Rapture of a renal angiomyolipoma and massive retroperitoneal hemorrhage, during pregnancy is rare and occasionally fatal. The association of this complication with pregnancy has been reported sporadically in the literature. We report a case of a 28 years old woman, in 33 week of her first pregnancy who came to our hospital complaining of abdominal pain in the right hemiabdomen, associated with nausea and vomiting. The ultrasound and MRI (Magnetic resonance imaging) scan showed a 7 x 7 x 5 cm mass suggestive of angiomyolipoma in the right kidney, with evidence of retroperitoneal bleeding right perirenal and intrarenal haematoma. Given the size of the tumor, presence of symptoms and hemodynamic instability of the patient right nephrectomy was performed, following emergency caesarean delivery. The histological study of the resected mass revealed the presence of angiomyolipoma. In conclusion, it seems that these tumours show a greater growth index in pregnant women. Angiomyolipoma with spontaneous bleeding during pregnancy is a dangerous condition that may cause mortality in the mother and fetus, but elective, simultaneous cesarean section and radical nephrectomy can be performed. We have also done a review of the literature focusing on its management and its relationship with pregnancy.


[Proposal for a national system of pacemaker patient registration. Goals, technical aspects and cost analysis (author's transl)].
Based on a personal experience with computer logging of pacemaker data, the Authors describe a system for a nation-wide registration of pacemaker patients. The advantages of such registration are stressed. The way of collecting and storing data, the interactions between pacemaker centers and the central registration system, the registration card, the estimated costs are discussed in detail.


Human urine-derived stem cells can be induced into osteogenic lineage by silicate bioceramics via activation of the Wnt/β-catenin signaling pathway.
Human urine-derived stem cells (USCs) have great application potential for cytotherapy as they can be obtained by non-invasive and simple methods. Silicate bioceramics, including calcium silicate (CS), can stimulate osteogenic differentiation of stem cells. However, the effects of silicate bioceramics on osteogenic differentiation of USCs have not been reported. In this study, at first, we investigated the effects of CS ion extracts on proliferation and osteogenic differentiation of USCs, as well as the related mechanism. CS particles were incorporated into poly (lactic-co-glycolic acid) (PLGA) to obtain PLGA/CS composite scaffolds. USCs were then seeded onto these scaffolds, which were subsequently transplanted into nude mice to analyze the osteogenic differentiation of USCs and mineralization of extracellular matrix formed by USCs in vivo. The results showed that CS ion extracts significantly enhanced cell proliferation, alkaline phosphatase (ALP) activity, calcium deposition, and expression of certain osteoblast-related genes and proteins. In addition, cardamonin, a Wnt/β-catenin signaling inhibitor, reduced the stimulatory effects of CS ion extracts on osteogenic differentiation of USCs, indicating that the observed osteogenic differentiation of USCs induced by CS ion extracts involves Wnt/β-catenin signaling pathway. Furthermore, histological analysis showed that PLGA/CS composite scaffolds significantly enhanced the osteogenic differentiation of USCs in vivo. Taken together, these results suggest the therapeutic potential of combining USCs and PLGA/CS scaffolds in bone tissue regeneration.


Endolymphatic sac tumor with von Hippel-Lindau disease: report of a case with analysis of von Hippel-Lindau gene and review.
Endolymphatic sac tumors (ELSTs) are very rare and locally aggressive low-grade neoplasm of endolymphatic system origin, which are associated with von Hippel-Lindau (VHL) disease. Evidence suggests that the specific VHL alteration influences the phenotype. Because of the rarity of ELSTs, only a small number of cases have been subjected to molecular genetic analysis. The correlation between ELSTs and the genotype of VHL remains unclear. Herein, we reported a case of ELST with VHL gene analysis who presented with a family history of VHL disease. The radiologic, histologic, and immunohistochemical features of the tumor were typical of ELST. Using the polymerase chain reaction-single-strand conformation polymorphism sequencing techniques, a germline mutation was identified as IVS1 + 1G→A at position 553 + 1. The mutation found in this case has not been previously reported in ELSTs. It is hoped that the study would contribute to a better understanding of ELSTs and the correlation between ELSTs and the genotype of VHL.


Cyclic AMP-associated shape change in mesangial cells and its reversal by prostaglandin E2.
The mesangial cell is a glomerular cell type with smooth muscle-like (contractile) properties. The responses evoked in cultured mesangial cells by catecholamines were examined in the presence or absence of prostaglandin E2 (PGE2) with or without a phosphodiesterase inhibitor. Exposure to 10(-4) M norepinephrine, epinephrine, or isoproterenol elevated intracellular cyclic AMP (cAMP) levels in mesangial cells (25th to 30th passages) nearly threefold. If isobutylmethylxanthine (MIX) was also included, the hormones caused marked further increases in cAMP (after a 20-min incubation, control with MIX, 64.2 +/- 5.2 pmoles/mg protein; 10(-4) M norepinephrine, 4266 +/- 284 pmoles/mg protein; 10(-4) M epinephrine, 5812 +/- 173 pmoles/mg protein; and 10(-4) M isoproterenol, 3136 +/- 114 pmoles/mg protein). Under both of these circumstances (that is, catecholamines with or without MIX) greater than 50% of the cells underwent a change in shape (that is, had a round cell body with long, thin tapered processes). The cAMP and shape change response was independent of extracellular calcium ions and appeared to be due to beta-adrenergic stimulation. Isoproterenol with MIX stimulated an alteration in morphology and cAMP production at concentrations of 10(-4) M to 10(-9) M. Within 10 min following beta-adrenergic stimulation (10(-4) M isoproterenol plus MIX) cAMP was maximum; at this time a shape change was first evident. Eighty-five to one hundred percent of the cells had undergone a shape change by 40 min. Dibutyryl cAMP (10(-3) M) also induced a shape change in cultured mesangial cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Single-Run Catalysis and Kinetic Control of Human Telomerase Holoenzyme.
Genome stability in eukaryotic cells relies on proper maintenance of telomeres at the termini of linear chromosomes. Human telomerase holoenzyme is required for maintaining telomere stability in a majority of proliferative human cells, making it essential for control of cell division and aging, stem cell maintenance, and development and survival of tumor or cancer. A dividing human cell usually contains a limited number of active telomerase holoenzymes. Recently, we discovered that a human telomerase catalytic site undergoes catalysis-dependent shut-off and an inactive site can be reactivated by cellular fractions containing human intracellular telomerase-activating factors (hiTAFs). Such ON-OFF control of human telomerase activity suggests a dynamic switch between inactive and active pools of the holoenzymes. In this review, we will link the ON-OFF control to the thermodynamic and kinetic properties of human telomerase holoenzymes, and discuss its potential contributions to the maintenance of telomere length equilibrium. This treatment suggests probabilistic fluctuations in the number of active telomerase holoenzymes as well as the number of telomeres that are extended in a limited number of cell cycles, and may be an important component of a fully quantitative model for the dynamic control of telomerase activities and telomere lengths in different types of eukaryotic cells.


Comments of Connecticut State Medical Society concerning proposed practitioner fee schedule regulations before the Workers' Compensation Commission 23 September 1993.
The Connecticut General Assembly passed legislation last session reforming the workers' compensation system. One provision of that legislation mandated the establishment of a medical fee schedule for the workers' compensation system by 1 October 1993. In response to that mandate, Workers' Compensation Commission Chairman Jesse Frankl relied upon the work and advice of his Medical Fee Advisory Group, of which the Connecticut State Medical Society was a member, and published proposed regulations for a medical fee schedule in the 31 August 1993 Connecticut Law Journal. A public hearing on those regulations was held on 23 September 1993. The following is testimony submitted by the Connecticut State Medical Society at that public hearing in support of those regulations.


Leiomyosarcoma of Mandible: A Diagnostic Dilemma; Case Report and Review of Literature.
Leiomyosarcoma and its pleomorphic variant are rare entities in the head and neck region. Since they usually present as slow growing, discrete firm, and non-ulcerated painless mass, they seem to be deceptively benign and are thus misdiagnosed. Histopathological and immunohistochemical studies are the ways of getting a definitive diagnosis. Till date surgery has been the primary treatment but effectiveness of radiotherapy/chemotherapy is still questionable. Here, we describe a case of leiomyosarcoma in mandible along with discussion about the ways of diagnosis, its differentiation with its pleomorphic variant and their managements.


Relation of Obesity to Outcomes of Hospitalizations for Atrial Fibrillation.
Obesity has been linked with increased incidence of atrial fibrillation (AF), but impact of presence of obesity on outcomes of hospitalizations for AF has not been investigated. We used the National Inpatient Sample database 2010 to 2014 to identify all adult hospitalizations aged ≥18years with a primary diagnosis of AF. Obese patients were identified using the co-morbidity variable for obesity, as defined in National Inpatient Sample databases. Multivariable logistic regression was used to compare in-hospital outcomes (mortality, acute stroke events) between obese and non-obese patients with AF. Of 431, 734 hospitalizations for AF, 66,138 (15.3%) were obese. Obese patients were younger and more likely to be African-Americans compared with non-obese patients. Despite being younger, obese patients had significantly higher prevalence of cardiovascular co-morbidities such as hypertension, diabetes mellitus, dyslipidemia, smoking, heart failure, and chronic renal failure (p <0.001 for all). After multivariate risk-adjustment, obese patients had a lower observed in-hospital mortality (0.5% vs 1.0%; unadjusted odds ratio = 0.52, 95% confidence interval [CI] 0.46 to 0.58, p <0.001; adjusted odds ratio = 0.83, 95% CI 0.73 to 0.94, p <0.001) and acute stroke events (0.4% vs 0.7%, unadjusted odds ratio = 0. 65, 95% CI 0.57 to 0.73, p < 0.001; adjusted odds ratio = 0.82, 95% CI 0.72 to 0.94) compared with non-obese patients. In conclusion, in this large retrospective analysis of an unselected nationwide cohort of patients hospitalized for AF, obese patients demonstrated lower risk-adjusted odds of in-hospital mortality and stroke events, consistent with an "obesity paradox."


Predicting tigecycline susceptibility in multidrug-resistant Klebsiella species and Escherichia coli strains of environmental origin.
Tigecycline (TGC) is an important antimicrobial agent used as a last resort for difficult-to-treat infections mainly caused by carbapenem-resistant Enterobacteriaceae, but TGC-resistant strains are emerging, raising concerns. In this study, 33 whole-genome characterized multidrug-resistant (MDR) strains (Klebsiella species and Escherichia coli) positive mainly to mcr-1, bla, and/or qnr from the environment were investigated for TGC susceptibility and mutations in TGC resistance determinants, predicting a genotype-phenotype relationship. TGC minimum inhibitory concentrations (MICs) of Klebsiella species and E. coli ranged from 0.25 to 8 and 0.125 to 0.5 mg/L, respectively. In this context, KPC-2-producing Klebsiella pneumoniae ST11 and Klebsiella quasipneumoniae subsp. quasipneumoniae ST4417 strains were resistant to TGC, while some E. coli strains of ST10 clonal complex positive for mcr-1 and/or blaCTX-M exhibited reduced susceptibility to this antimicrobial. Overall, neutral and deleterious mutations were shared among TGC-susceptible and TGC-resistant strains. A new frameshift mutation (Q16stop) in RamR was found in a K. quasipneumoniae strain and was associated with TGC resistance. Deleterious mutations in OqxR were identified in Klebsiella species and appear to be associated with decreased susceptibility to TGC. All E. coli strains were determined as susceptible, but multiple point mutations were identified, highlighting deleterious mutations in ErmY, WaaQ, EptB, and RfaE in strains exhibiting decreased susceptibility to TGC. These findings demonstrate that resistance to TGC is not widespread in environmental MDR strains and provide genomic insights about resistance and decreased susceptibility to TGC. From a One Health perspective, the monitoring of TGC susceptibility should be constant, improving the genotype-phenotype relationship and genetic basis.


Characterization of an iPSC line NCHi006-A from a patient with hypoplastic left heart syndrome (HLHS).
Hypoplastic left heart syndrome (HLHS) is a severe congenital heart defect characterized by underdeveloped structures on the left side of the heart, including hypoplasia of the left ventricle and stenosis or atresia of the aortic and mitral valves. Here, we generated an iPSC line from the peripheral blood mononuclear cells of a male patient with HLHS through Sendai virus-mediated transfection of 4 Yamanaka factors. This iPSC line exhibited normal morphology, expressed pluripotency markers, had a normal karyotype, and could differentiate into cells of three germ layers. This iPSC line can be used for studying cellular and developmental etiologies of HLHS.


Change of the protein composition of the thyroid colloid during treatment with propylthiouracil and thyroxine: a microgel electrophoretic study of single rat thyroid follicles.
Prophylthiouracil and thyroxine were given daily to rats for 4 weeks. Samples of colloid were collected in vivo from the superficial thyroid follicles during this period and their protein composition was analysed by gel electrophoresis. It was observed that the aggregates of thyroglobulin, i.e., the 27-S thyroid iodoprotein and the heavier fractions, were reduced to 50% after 1 week and were almost absent after 2 weeks. A faster migrating thyroglobulin fraction was observed in the samples of colloid and in the homogenate of the whole gland after 48 h of treatment. During the following period of treatment there was an increase in the relative amount of the faster migrating thyroglobulin fraction compared to 19-S thyroglobulin in the colloid, the former comprising approx. 75% of the globulins after 4 weeks, It can be concluded that propylthiouracil inhibits the formation of the 27-S iodoprotein and that a structurally altered and iodine-poor thyroglobulin fraction is accumulated in the follicle lumen.


COMPARATIVE EVALUATION OF NO-SCALPEL VASECTOMY AND STANDARD INCISIONAL VASECTOMY.
No-scalpel vasectomy employs a refined method of dissection and delivery of the vas deferens. We compared no-scalpel vasectomy with standard incisional vasectomy in 176 patients over a 33 month period. The haemorrhage rate was 1.08 per cent for no-scalpel vasectomy compared with 11.9 per cent for standard vasectomy (p < 0.005). The infection rate was 3.26 per cent for no-scalpel vasectomy as against 14.28 per cent for standard vasectomy (p < 0.01). There was a 37.5 per cent reduction in operating time and a substantial reduction in pain during and after the procedure when no-scalpel vasectomy was performed and also there was no failure of vasectomy. No-scalpel vasectomy is a satisfactory alternative to standard vasectomy with fewer complications and increased patient acceptability.


2-(2-Selenocyanic acid ethyl ester)-1H-benz[de] isoquinoline-1,3-(2H)-dione, synthesis photophysics and interaction with bovine serum albumin: a spectroscopic approach.
The compound 2-(2-selenocyanic acid ethyl ester)-1H-benz[de] isoquinoline-1,3-(2H)-dione (SEBID), a ubiquitous, bioactive naphthalimide derivative is expected to possess an anticancer, anti-tumor and other important therapeutic activities of significant potency with low systematic toxicity. In this paper, the synthesis of the compound, photophysics of the newly prepared naphthalimide derivative and its interaction with model transport protein Bovine serum albumin (BSA) have been reported using the absorption and steady state fluorescence spectroscopic techniques exploiting the intrinsic fluorescence emission properties of BSA as a probe. Interaction of this organoselenium compound in different dioxane-water mixtures with increase in the polarity of the medium has been studied spectroscopically. Interaction of SEBID with BSA leads to a dramatic decrease in the fluorescence intensity of BSA, which suggests the binding of SEBID with the tryptophan residue of BSA. Furthermore, different thermodynamic parameters for SEBID-BSA interaction have been calculated. Rationalization of the data has been attempted, particularly in relation to prospective applications in the biomedical research.


Quantitative Proteomics Reveals the Mechanism of Silver Nanoparticles against Multidrug-Resistant Pseudomonas aeruginosa Biofilms.
The decline of clinically effective antibiotics has made it necessary to develop more effective antimicrobial agents, especially for refractory biofilm-related infections. Silver nanoparticles (AgNPs) are a new type of antimicrobial agent that can eradicate biofilms and reduce bacterial resistance, but its anti-biofilm mechanism has not been elucidated. In this study, we investigated the molecular mechanism of AgNPs against multidrug-resistant Pseudomonas aeruginosa by means of anti-biofilm tests, scanning electron microscopy (SEM), and tandem mass tag (TMT)-labeled quantitative proteomics. The results of anti-biofilm tests demonstrated that AgNPs inhibited the formation of P. aeruginosa biofilm and disrupted its preformed biofilm. SEM showed that when exposed to AgNPs, the structure of the P. aeruginosa biofilm was destroyed, along with significant reduction of its biomass. TMT-labeled quantitative proteomic analysis revealed that AgNPs could defeat the P. aeruginosa biofilm in multiple ways by inhibiting its adhesion and motility, stimulating strong oxidative stress response, destroying iron homeostasis, blocking aerobic and anaerobic respiration, and affecting quorum sensing systems. Our findings offer a new insight into clarifying the mechanism of AgNPs against biofilms, thus providing a theoretical basis for its clinical application.


"Kiss-and-run" glutamate secretion in cultured and freshly isolated rat hippocampal astrocytes.
Under physiological conditions, astrocytes not only passively support and nourish adjacent neurons, but also actively modulate neuronal transmission by releasing "glial transmitters," such as glutamate, ATP, and D-serine. Unlike the case for neurons, the mechanisms by which glia release transmitters are essentially unknown. Here, by using electrochemical amperometry and frequency-modulated single-vesicle imaging, we discovered that hippocampal astrocytes exhibit two modes of exocytosis of glutamate in response to various stimuli. After physiological stimulation, a glial vesicle releases a quantal content that is only 10% of that induced by nonphysiological, mechanical stimulation. The small release event arises from a brief (approximately 2 ms) opening of the fusion pore. We conclude that, after physiological stimulation, astrocytes release glutamate via a vesicular "kiss-and-run" mechanism.


Economic and Clinical Burden of Managing Sickle Cell Disease with Recurrent Vaso-Occlusive Crises in the United States.
The aim of this study was to describe the clinical complications, treatment use, healthcare resource utilization (HCRU), and costs among patients with sickle cell disease (SCD) with recurrent vaso-occlusive crises (VOCs) in the US. Merative MarketScan Databases were used to identify patients with SCD with recurrent VOCs from March 1, 2010, to March 1, 2019. Inclusion criteria were ≥ 1 inpatient or ≥ 2 outpatient claims for SCD and ≥ 2 VOCs per year in any 2 consecutive years after the first qualifying SCD diagnosis. Individuals without SCD in these databases were used as matched controls. Patients were followed for ≥ 12 months, from their second VOC in the 2nd year (index date) to the earliest of inpatient death, end of continuous enrollment in medical/pharmacy benefits, or March 1, 2020. Outcomes were assessed during follow-up. In total, 3420 patients with SCD with recurrent VOCs and 16,722 matched controls were identified. Patients with SCD with recurrent VOCs had a mean of 5.0 VOCs (standard deviation [SD] = 6.0), 2.7 inpatient admissions (SD 2.9), and 5.0 emergency department visits (SD 8.0) per patient per year during follow-up. Compared to matched controls, patients with SCD with recurrent VOCs incurred higher annual ($67,282 vs. $4134) and lifetime ($3.8 million vs. $229,000 over 50 years) healthcare costs. Patients with SCD with recurrent VOCs experience substantial clinical and economic burden driven by inpatient costs and frequent VOCs. There is a major unmet need for treatments that alleviate or eliminate clinical complications, including VOCs, and reduce healthcare costs in this patient population.


Variability in the immune system: of vaccine responses and immune states.
System-wide approaches are now being applied to study vaccine responses, whose mechanisms of action, and failure, are not well understood. These works have repeatedly shown vaccine response to be an orchestrated process involving multiple arms of immunity most noticeable sensing and innate components. Prediction of vaccine responses based on system-wide measures is achievable, but challenges remain for robust population wide predictions based only on pre-vaccination measures, especially in partially efficacious vaccines such as influenza. This is especially true in older adults, who are often less responsive to vaccination and exhibit high level of variation compared to young in many components of immunity. Despite this increase in variation, most of the studies on aging use group averages of immune phenotypes to model immune system behavior. Using systems approaches, it is possible to exploit this variation to form distinguishable clusters of phenotypes within and across individuals to discover underlying immune states.


External validation study of a clinical decision aid to reduce unnecessary antibiotic prescriptions in women with acute cystitis.
Empirical prescribing of antibiotics to women with symptoms of acute cystitis prior to culture results is common, but subsequent culture results are often negative. A clinical decision aid for prescribing decisions in acute cystitis was previously developed that could reduce these unnecessary antibiotic prescriptions but has not been validated. This study sought to validate this decision aid for empirical antibiotic prescribing decisions in a new cohort of women with suspected acute cystitis. External validation study of a clinical decision aid in 397 women with symptoms of acute cystitis, involving 230 Canadian family practitioners across Canada between 2009 and 2011. The sensitivity and specificity of the decision aid compared to a gold standard positive urine culture (defined as ≥102 cfu/ml (≥105 CFU/L)) was determined, and compared with physician management, and the earlier development cohort study estimates. Other outcomes assessed were total antibiotic prescriptions, unnecessary antibiotics for negative urine cultures, and recommendations for urine culture testing. Chi-square tests were used for unpaired comparisons, adjusted for physician clustering. McNemar's test was used for paired comparisons. There were 245/397 (61.7%) positive urine cultures. The cystitis aid sensitivity was 202/245 (82.5%, 95% Confidence Interval (CI)) = 77.1%, 86.8%), compared to 167/208 (80.3%) in the previous development cohort (p = 0.54), and 239/245 (97.6%) by family physicians in the current study (p < 0.001). Specificity was low for physicians (10/152, 6.6%) compared to the decision aid (54/152, 35.5%; p < 0.001, resulting in more antibiotic prescriptions by physicians (381/397, 96.0%) than would occur with decision aid recommendations (300/397, 75.6%, p < 0.001). Unnecessary antibiotic prescriptions where urine cultures were negative would be reduced an absolute 11.1% with cystitis aid recommendations (98/397, 24.7%) compared to usual physician care (142/397, 35.8%; p = 0.001). Urine cultures would also be reduced (97/397, 24.4% decision aid vs 351/397, 88.4% physicians; p < 0.001). A 3-item clinical decision aid demonstrated reproducible accuracy in two cohorts of women with acute cystitis symptoms. Clinically important reductions in total and unnecessary antibiotic use, as well as urine culture testing, could result with routine clinical use compared to current empirical physician management practices.


Ostial PV isolation: is there a role for three-dimensional mapping?
Pulmonary vein (PV) isolation by elimination of spike potentials has been reported to cure drug refractory atrial fibrillation. Because of the heterogenous morphology of the PVs, sequential electroanatomic reconstruction of the PVs was performed in 39 patients (group A), who underwent subsequent PV isolation by interruption of all conductive myocardial fibers by distinct RF current applications using a "lasso" approach. In group B (157 patients), only biplane two-dimensional fluoroscopy was performed to guide the diagnostic and the ablation catheters. After reprocedures (in 7% of patients in group A and 22% of group B), which depicted a recurrence of a spike potential inside or at the ostium of >1 previously isolated PV in all restudied patients, stable sinus rhythm was documented in 69% of patients in group A and 60% of patients in group B. Reasons for the relapse of the previously eliminated spike potentials include a temporary ablation effect and a too distal interruption of the conducting myocardial fiber. Detailed knowledge of the individual three-dimensional morphology enhanced the clinical success rate of PV isolation but is time-consuming using CARTO (8.0 +/- 1.7 vs 5.0 +/- 1.6, P < 0.001). Further technical improvement to fuse the individual three-dimensional anatomy and the electrophysiological markers to a composed "electroanatomic" map may overcome this limitation in the future.


Relationship between alcohol and nutrient intakes and blood status indices of older people living in the UK: further analysis of data from the National Diet and Nutrition Survey of people aged 65 years and over, 1994/5.
To examine the relationships between alcohol consumption and a range of nutrient intakes and blood status indices in older people. National Diet and Nutrition Survey: cross-sectional survey of nationally representative sample of people aged 65 years or over. Mainland Britain during 1994/5. 1198 people (623 males, 575 females) aged 65 years or over, of whom 925 were living in private households and 273 were living in institutions. Intermediate alcohol consumption (particularly 0.1-14 units week(-1); 1 unit = 8 g) derived from a 4-day diet diary or a 12-month recall questionnaire, was associated with higher intakes of vitamins C, E, B1, iron, calcium, energy from food, carbohydrate and non-starch polysaccharides than heavy alcohol consumption (28 + units week(-1)) or abstinence, after adjustment for a number of factors (age, sex, domicile, social class, cigarette smoking, self-reported health, grip strength and total energy intake). Intermediate alcohol consumption was also associated with higher blood concentrations (independent of intake) of vitamin C, beta-cryptoxanthin, lutein and calcium, with the lowest concentrations being found in heavy alcohol users. The lowest concentrations of serum ferritin were found in light drinkers and the highest levels in heavier alcohol drinkers. Alcohol consumption ranging from 0.1 to >28 units week(-1) was directly correlated with intakes of B vitamins, total energy and fat, with blood concentrations (independent of intake) of lycopene, high density lipoprotein (HDL)-cholesterol, plasma pyridoxal phosphate and retinol, and with blood pressure and grip strength. Compared with abstinence and heavy drinking, light to moderate alcohol consumption in older people is associated with higher intakes of certain nutrients, and higher blood concentrations (independent of intake) of some micronutrient status indices, including antioxidants. The explanation for the latter associations remains unclear and further investigation is recommended. Heavier alcohol consumption is associated with both beneficial and adverse effects with respect to nutrient intakes and health status.


Influences of gender in metabolic syndrome and its components among people living with HIV virus using antiretroviral treatment in Hawassa, southern Ethiopia.
Data regarding the influences of gender in metabolic syndrome (MetS) among patients using antiretroviral treatment (ART) in Ethiopia is scarce. The aim of this study was to assess the influences of gender in MetS and its components among HIV-infected patients receiving ART. A cross-sectional study was conducted between February 2012 and April 2013. Data on demographic, clinical and anthropometric characteristics were collected from 185 HIV patients using ART. Glucose and lipid profiles were measured from overnight fast blood. The International Diabetes Federation (IDF) and United States national cholesterol education program: adult treatment (US NCEP-ATP) panel III criteria were used to define MetS. A total number of 185 (36.8% males and 63.2% females) participants were recruited in this study. The overall prevalence of MetS was 24.3 and 17.8%, diagnosed using IDF and NCEP-ATP criteria respectively. Using IDF criteria, MetS was significantly higher in females compared to males (33.3 vs. 8.8%; p = <0.0001) respectively. Low HDL-c and central obesity were significantly higher MetS components in female compared to males (p = 0.003); and (p = <0.0001, using IDF and NCEP-ATP criteria) respectively. BMI >25 kg/m(2) was significantly associated with MetS in both IDF and NCEP-ATP criteria: unadjusted (UOR) and adjusted odds ratio (AOR) with 95% CI were 3.0 (1.3-6.5) and 3.8 (1.5-9.8); as well as 3.2 (1.4-7.4) and 3.4 (1.4-7.4) respectively. Furthermore age >40 years was significantly associated with MetS using NCEP-ATP: UOR and AOR (95% CI) were 3.1 (1.2-8.3), and 3.8 (1-13.70) respectively. Comprehensive medical care approach including with MetS components are a crucial instruments in order to minimize the risk of developing cardiovascular diseases in HIV-infected patients using ART.


Exploring the application of stem cells in tendon repair and regeneration.
To conduct a systematic review of the current evidence for the effects of stem cells on tendon healing in preclinical studies and human studies. A systematic search of the PubMed, CINAHL (Cumulative Index to Nursing and Allied Health Literature), Cochrane, and Embase databases was performed for stem cells and tendons with their associated terminology. Data validity was assessed, and data were collected on the outcomes of trials. A total of 27 preclinical studies and 5 clinical studies met the inclusion criteria. Preclinical studies have shown that stem cells are able to survive and differentiate into tendon cells when placed into a new tendon environment, leading to regeneration and biomechanical benefit to the tendon. Studies have been reported showing that stem cell therapy can be enhanced by molecular signaling adjunct, mechanical stimulation of cells, and the use of augmentation delivery devices. Studies have also shown alternatives to the standard method of bone marrow-derived mesenchymal stem cell therapy. Of the 5 human studies, only 1 was a randomized controlled trial, which showed that skin-derived tendon cells had a greater clinical benefit than autologous plasma. One cohort study showed the benefit of stem cells in rotator cuff tears and another in lateral epicondylitis. Two of the human studies showed how stem cells were successfully extracted from the humerus and, when tagged with insulin, became tendon cells. The current evidence shows that stem cells can have a positive effect on tendon healing. This is most likely because stem cells have regeneration potential, producing tissue that is similar to the preinjury state, but the results can be variable. The use of adjuncts such as molecular signaling, mechanical stimulation, and augmentation devices can potentially enhance stem cell therapy. Initial clinical trials are promising, with adjuncts for stem cell therapy in development.


Recent development of ionic liquid stationary phases for liquid chromatography.
Based on their particular physicochemical characteristics, ionic liquids have been widely applied in many fields of analytical chemistry. Many types of ionic liquids were immobilized on a support like silica or monolith as stationary phases for liquid chromatography. Moreover, different approaches were developed to bond covalently ionic liquids onto the supporting materials. The obtained ionic liquid stationary phases show multi-mode mechanism including hydrophobic, hydrophilic, hydrogen bond, anion exchange, π-π, and dipole-dipole interactions. Therefore, they could be used in different chromatographic modes including ion-exchange, RPLC, NPLC and HILIC to separate various classes of compounds. This review mainly summarizes the immobilized patterns and types of ionic liquid stationary phases, their retention mechanisms and applications in the recent five years.


Inception loops discover what excites neurons most using deep predictive models.
Finding sensory stimuli that drive neurons optimally is central to understanding information processing in the brain. However, optimizing sensory input is difficult due to the predominantly nonlinear nature of sensory processing and high dimensionality of the input. We developed 'inception loops', a closed-loop experimental paradigm combining in vivo recordings from thousands of neurons with in silico nonlinear response modeling. Our end-to-end trained, deep-learning-based model predicted thousands of neuronal responses to arbitrary, new natural input with high accuracy and was used to synthesize optimal stimuli-most exciting inputs (MEIs). For mouse primary visual cortex (V1), MEIs exhibited complex spatial features that occurred frequently in natural scenes but deviated strikingly from the common notion that Gabor-like stimuli are optimal for V1. When presented back to the same neurons in vivo, MEIs drove responses significantly better than control stimuli. Inception loops represent a widely applicable technique for dissecting the neural mechanisms of sensation.


[Urodynamics in female stress urinary incontinence].
Urodynamic studies are essential in order to rule out the existence of vesical instabilities. In our opinion, the urethral outlines are not always revealing and demand, apart from a highly planned technique, a critical and prudent assessment of the results.


Immunotargeting of insulin reactive CD8 T cells to prevent diabetes.
Insulin is one of the earliest targeted autoantigens in the immune destruction of insulin-producing beta cells by autoreactive CD4 and CD8 T cells in type 1 diabetes. In this study, we used Non-obese diabetic (NOD) transgenic T cells engineered to express MHC class I-insulin peptide complexes linked to a T cell activation component (InsCD3-ζ), to target insulin-reactive CD8 T cells. We showed that activated, but not naïve, InsCD3-ζ CD8 T cells killed diabetogenic insulin-reactive CD8 target cells in vitro, inducing antigen-specific cell death mediated via both the release of perforin and the Fas-Fas ligand pathway. In vivo, InsCD3-ζ CD8 T cells migrated to the pancreatic lymph nodes of NOD mice after adoptive transfer. Concomitant with this, infiltration of CD8 T cells was also reduced in the pancreatic islets. Finally, in vivo, we showed that diabetes induced by adoptive transfer of insulin-reactive T cells was reduced following injection of activated InsCD3-ζ CD8 T cells. Furthermore, young NOD mice injected with InsCD3-ζ CD8 T cells developed a lower incidence and delayed onset of diabetes. Thus, using this novel system we have demonstrated that InsCD3-ζ CD8 T cells can directly kill insulin-reactive CD8 T cells in vitro and by targeting insulin-specific CD8 T cells early in the course of disease alter the progression of spontaneous diabetes in vivo in NOD mice.


Modelling autocatalytic networks with artificial microbiology.
Cells can usefully be equated to autocatalytic networks that increase in mass and then divide. To begin to model relationships between autocatalytic networks and cell division, we have written a program of artificial chemistry that simulates a cell fed by monomers. These monomers are symbols that can be assembled into linear (non-branched) polymers to give different lengths. A reaction is catalysed by a particular polymer or 'enzyme' that may itself be a reactant of that reaction (autocatalysis). These reactions are only studied within the confines of the 'cell' or 'reaction chamber'. There is a flux of material through the cell and eventually the mass of polymers reaches a threshold at which we analyse the cell. Our results indicate a similarity between the connectivity of the reaction network and that of real metabolic networks. Developing the model will entail attributing increased probabilities of reactions to polymers that are colocalised to evaluate the consequences of the dynamics of large assemblies of diverse molecules (hyperstructures) and of cell division.


Kinetics of meiotic maturation in oocytes from unstimulated ovaries and duration of pronucleus presence and preimplantation development.
To evaluate the meaning of meiotic maturation kinetics and duration of pronucleus presence (DPP) for parthenogenetic activation outcome. Retrospective study. University hospital. Eight patients with endometrioid adenocarcinoma and 65 patients who underwent in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI). After collection of oocytes from nonstimulated ovaries of patients with endometrioid adenocarcinoma, in vitro maturation (IVM) and parthenogenetic activation performed with time-lapse imaging; after ICSI, embryos similarly incubated with time-lapse imaging. Timing of the release of the first polar body (fPB), DPP, and developmental stage with IVM and parthenogenetic activation; after ICSI, assessment of DPP and preimplantation developmental stage. With IVM, 55.2% of oocytes matured; 53.1% of fPBs were released within 24 hours, and 46.9% of fPBs were released after 24 hours. Regarding developmental stage, oocytes that released fPB later during IVM tended to develop more than oocytes that released the fPB within 24 hours. For embryos from parthenogenetic activation the DPP was statistically significantly shorter than the DPP of embryos from ICSI. With ICSI, the DPP was statistically significantly shorter in embryos that developed to ≥8 cells than embryos whose final development included ≤7 cells. The development rate in parthenogenetic activation was statistically significantly lower than that in ICSI. Embryo development is negatively affected by DPP that is too short or too long. When the DPP was short with parthenogenetic activation, embryo development did not proceed, indicating that DPP is an important determinant of parthenogenetic activation outcomes as with the timing of fPB release.


The impact of persistent pediatric obstructive sleep apnea on the Quality of Life of Patients' families.
To (1) quantify the quality of life (QOL) for families of children affected by persistent obstructive sleep apnea (OSA), and (2) assess factors contributing to the negative impact of OSA on families. Prospective case series in a multidisciplinary upper airway center at a tertiary pediatric institution. Our study included patients with persistent OSA referred to our clinic from 2014 to 2016. Both patients and their families completed validated questionnaires assessing QOL and OSA symptoms, including the Family Impact Questionnaire (FIQ), Pediatric Sleep Questionnaire, the Pediatric Quality of Life Inventory, the OSA-18, and the Epworth Sleepiness Scale. Families of 67 patients were included. The mean patient age was 12.5 (95%CI 11.9-13.1); 23 (34.3%) were female, and the mean BMI percentile was 80.2 (95%CI 73.6-86.8). The single most common comorbidity was Down syndrome (45.6%). The mean obstructive apnea-hypopnea index was 9.7 ± 10.3 events/hour. A significant association exists between OSA severity and 18 FIQ negative subscore (P < 0.001). Financial impact was the primary negative concern for parents of patients with OSA compared to those without OSA (P = 0.03). There were no other significant differences between those with and without OSA. There was a significant correlation between persistent OSA severity and the FIQ negative impact of disease score on patients' families. Concern regarding financial burden was more common for families of children with OSA than for those without OSA. This suggests that targeted interventions, particularly regarding the financial burden of persistent OSA diagnosis and management, may provide some relief to families.


Nitroimidazoles as anaerobic electron acceptors for xanthine oxidase.
The rates of anaerobic nitroreduction of 15 nitroimidazoles by xanthine-xanthine oxidase were measured. The compounds studied were mainly 2-nitroimidazoles, including misonidazole and analogues of potential value in cancer therapy; most 5-nitroimidazoles such as metronidazole and nimorazole reacted too slowly for measurement under the conditions used. Using 0.1 mM nitroimidazole, reduction rates varied between ca. 4 and 680 nmole min-1 U-1 xanthine oxidase at 37 degrees and pH 7.4. These rates were correlated with the reduction potential characterizing one-electron reduction to the nitro radical-anion, and compared with those for other nitroaryl compounds and with reduction by free flavin mononucleotide.


Structure-function relationships in the transposition protein B of bacteriophage Mu.
The B-protein of phage Mu, which is required for high frequency intermolecular transposition in vivo, shows ATPase activity in vitro, binds nonspecifically to DNA, and stimulates intermolecular strand transfer. To elucidate the structural bases for B-protein function, it was subjected to limited proteolysis with two different proteases, trypsin and chymotrypsin. The resulting fragments were mapped by amino acid sequencing. These data show that the B-protein is organized in two domains: an amino-terminal domain of 25 kDa and a carboxyl-terminal domain of 8-kDa. A fragment analogous to the amino-terminal domain, produced by deleting the 3' end of a cloned B gene, proved to be insoluble and had to be renatured after elution from a sodium dodecyl sulfate gel. The renatured protein retains ATP-binding activity and to a lesser extent the DNA-binding activity of the MuB protein, but is unable to hydrolyze ATP or function in transposition. We also show in this study that efficient DNA-strand transfer by the B-protein occurs even in the absence of a detectable ATPase activity or in the presence of adenosine 5'-O-(thio)triphosphate (ATP gamma S).


Feasibility of ultra-high-speed acquisition in xSPECT bone algorithm: a phantom study with advanced bone SPECT-specific phantom.
Although xSPECT Bone (xB) provides quantitative single-photon emission computed tomography (SPECT) high-resolution images, patients' burden remains high due to long acquisition time; therefore, this study aimed to investigate the feasibility of shortening the xB acquisition time using a custom-designed phantom. A custom-designed xSPECT bone-specific (xSB) phantom with simulated cortical and spongious bones was developed based on the thoracic bone phantom. Both standard- and ultra-high-speed (UHS) xB acquisitions were performed in a male patient with lung cancer. In this phantom study, SPECT was acquired for 3, 6, 9, 12, and 30 min. The clinical SPECT acquisition time per rotation was 9 and 3 min for standard and UHS, respectively. SPECT images were reconstructed using ordered subset expectation maximization with three-dimensional resolution recovery (Flash3D; F3D) and xB algorithms. Quantitative SPECT value (QSV) and coefficient of variation (CV) were measured using the volume of interests (VOIs) placed at the center of the vertebral body and hot sphere. A linear profile was plotted on the spinous process at the center of the xSB phantom; then, the full width at half maximum (FWHM) was measured. The standardized uptake value (SUV) and standard deviation from the first thoracic to the fifth lumbar vertebrae in clinical standard- and UHS-xB images were measured using a 1-cm3 VOI. The QSV of F3D images was underestimated even in large regions, whereas those of xB images were close to actual radioactivity concentration. The CV was similar or lower for xB images than that for F3D images but was not decreased with increasing acquisition time for both reconstruction images. The FWHM of xB images was lower than those of F3D images at all acquisition times. The mean SUV values from the first thoracic to fifth lumbar vertebrae for standard- and UHS-xB images were 6.73 ± 0.64 and 6.19 ± 0.87, respectively, showing a strong positive correlation. Results of this phantom study suggest that xB imaging can be obtained in only one-third of the acquisition time without compromising the image quality. The SUV of UHS-xB images can be similar to that of standard-xB images in terms of clinical interpretation.


Inhibition of proteolysis by the proform of eosinophil major basic protein (proMBP) requires covalent binding to its target proteinase.
By proteolytic cleavage of insulin-like growth factor binding proteins, the metalloproteinase pregnancy-associated plasma protein-A (PAPP-A) is able to control the biological activity of insulin-like growth factors. PAPP-A circulates in pregnancy as a proteolytically inactive complex, disulfide bound to the proform of eosinophil major basic protein (proMBP). We here demonstrate that co-transfection of mammalian cells with PAPP-A and proMBP cDNA results in the formation of a covalent PAPP-A/proMBP complex in which PAPP-A is inhibited. Formation of the complex also occurs when PAPP-A and proMBP synthesized separately are incubated. Complex formation was monitored by Western blotting, and by using an immunoassay specific for the complex. Using mutagenesis, we further demonstrate that the complex forms in a specific manner and depends on the presence of two proMBP cysteine residues. Mutated proMBP, in which Cys-51 and -169 are replaced by serine, is unable to form the covalent complex with PAPP-A. Of particular interest, such mutated proMBP further lacks the ability to inhibit PAPP-A. For the first time, this conclusively demonstrates that proMBP is a proteinase inhibitor. We further conclude that proMBP inhibits PAPP-A in an unusual manner, not paralleled by other proteinase inhibitors of our knowledge, which requires proMBP to be covalently bound to PAPP-A by disulfide bonds. ProMBP binding to PAPP-A most likely either abrogates substrate access to the active site of PAPP-A or induces a conformational change in the structure of PAPP-A, as we, by further mutagenesis, were able to exclude that the inhibitory mechanism of proMBP is based on a cysteine switch-like mechanism.


Pain management.
Postoperative pain management in the critically ill patient is a challenge for nurses. Knowing the basis of pain transmission and mechanisms of action of interventions can assist the critical care nurse in making clinical decisions regarding pain control for individual patients. There are a number of modalities available to treat postoperative pain including both pharmacologic and nonpharmacologic interventions. Techniques such as PCA not only can provide good analgesia, but allow the critically ill patient at least one aspect of control in the otherwise highly controlled environment of the critical care unit. Epidural or intrathecal analgesia, using either opioids or LAAs alone or in combination, provides excellent analgesic effect (with minimal side effects) and may improve patient outcomes. Nonpharmacologic techniques, unfortunately, are commonly overlooked as adjuncts to traditional analgesia routines because of the nature of the illness in the critically ill patient. Nonpharmacologic techniques of pain management have a place in the care of the critically ill when applied based on the assessment of an individual patient's needs and abilities to participate in his or her care. Ensuring optimal patient comfort can benefit critically ill patients and improve clinical outcomes.


Causes and control of filamentous growth in aerobic granular sludge sequencing batch reactors.
Poor long-term stability of aerobic granules developed in sequencing batch reactors (SBRs) remains a limitation to widespread use of aerobic granulation in treating wastewater. Filamentous growth has been commonly reported in aerobic granular sludge SBR. This review attempts to address the instability problem of aerobic granular sludge SBR from the perspective of filamentous growth in the system. The possible causes of filamentous growth are identified, including long retention times of solids, low substrate concentration in the liquid phase, high substrate gradient within the granule, dissolved oxygen deficiency in the granule, nutrient deficiency inside granule, temperature shift and flow patterns. Because of cyclic operation of aerobic granular sludge SBR and peculiarities of aerobic granules, various stresses can be present simultaneously and can result in progressive development of filamentous growth in aerobic granular sludge SBR. Overgrowth of filamentous bacteria under stress conditions appears to be a major cause of instability of aerobic granular sludge SBR. Specific recommendations are made for controlling filamentous growth.


Primaquine and lysosomotropic amines inhibit malaria sporozoite entry into human liver cells.
The binding and entry of Plasmodium berghei sporozoites to human hepatoma HepG2 cells is inhibited in a dose-dependent manner by primaquine, chloroquine and other lysosomotropic amines. The site of action of these agents appears to be the hepatoma cell itself, not the sporozoite. While this inhibitory effect of primaquine is rapidly reversible, the precise mechanism responsible for this effect is not presently known.


Effect of Chlorhexidine on the Bonding Effect of an Etch-and-Rinse Adhesive to Pretreatment Dentin.
The purpose of this study was to evaluate the effect of 2% chlorhexidine (CHX) on an etch-and rinse adhesive to dentin. Caries-free molars were selected and processed to expose a flat dentin surface. The specimens were bonded with a composite resin by an etch-and-rinse adhesive, which was pretreated with 2% CHX for 0, 15, 30, 45, and 60 seconds. The microtensile bond strength (MTBS) was evaluated before and after thermocycling. No significant differences were observed between groups before thermocycling. The 60-second CHX-pretreated group showed a significantly greater MTBS than the control group after thermocycling. The 2% CHX pretreatment could improve the bonding strength of the etch-and-rinse adhesive and slow down the aging progress of the bonding interface.


Tramped teats--clinical mastitis disease complex in tied cows. Environmental risk factors and interrelationships with other diseases.
The effects of stall length, manure system, type of bedding and calving disorders on incidence rates of tramped teats, udder injuries, and clinical mastitis in Swedish Red and White tied cows, as well as interrelationships between these 3 disorders were investigated. Data used consisted of 370 herds, for a total of 45,133 cow records. Cows in herds with liquid manure system were at higher risk of udder injuries and mastitis than in herds with solid system. Lower risk of both udder injuries and mastitis was found for cows in herds with short stall size (less than 180 cm) compared to herds with stalls of 205-219 cm length. It was suggested that other factors, such as slipperiness of the stall floor and presence and type of feeding barrier, must be considered when the effect of stall characteristics on tramped teats is investigated. In farms with solid manure system cut straw or sawdust bedding increased the risk of tramped teats and mastitis relative to uncut straw. Parity and calving disorders were important risk factors for udder health and their effect was greater in herds with liquid manure system. Tramped teats and udder injuries were the most serious risk factors for clinical mastitis.


The alterations of vascular smooth muscle reactivity in vitro by human chorionic gonadotrophin.
The study tests the hypothesis that human chorionic gonadotrophin (hCG) alters vascular smooth muscle reactivity by examining the effect of hCG administration on the contractility and relaxation of isolated vascular smooth muscle. Aortic rings from rats pre-treated with intraperitoneal administration of 5,000 I.U of hCG and control animals were contracted to phenylephrine, angiotensin II, CaCl2 and KCl. The experiments with phenylephrine were repeated with rings that were either de-endothelialized, incubated with L-NMMA, or incubated with calcium ionophore A23187. Aortic rings precontracted with phenylephrine were relaxed to acetylcholine (endothelium-dependent), sodium nitroprusside, hydralazine (endothelium-independent) or in the presence of A23187. The contractile responses of aortic rings from hCG-treated animals of phenylephrine, angiotensin II, CaCl2 and KCl were significantly attenuated. This effect was not reversed by pre-treatment with L-NMMA or by de-endothelialisation. In aortic rings from hCG-treated animals, there was almost total inhibition of acetylcholine-induced relaxation, but unaltered relaxation responses to sodium nitroprusside and hydralazine. The inhibitory effects of hCG-treatment on both the contraction and relaxation responses were either fully or partially reversed in the presence of calcium ionophore A23187. These observations suggest that hCG attenuates both contractile and endothelium-dependent relaxation responses by a mechanism which involves inhibition of extracellular calcium ion influx and may indicate a new role for the hormone in the altered vascular responses of both normal and abnormal pregnancies.


Vanadate protects human neuroblastoma SH-SY5Y cells against peroxynitrite-induced cell death.
We investigated the effect of vanadate, a tyrosine phosphatase inhibitor, on cell death induced by peroxynitrite in human neuroblastoma SH-SY5Y cells. Vanadate prevented cell death induced by 3-morpholinosydnonimine (SIN-1), a peroxynitrite donor; whereas SIN-1-induced cell death was not prevented by neither okadaic acid, an inhibitor of serine/threonine phosphatases 1 and 2A, nor cyclosporin A, an inhibitor of serine/threonine phosphatase 2B. Vanadate did not prevent cell death induced by N-ethyl-2-(1-ethyl-hydroxy-2-nitrosohydrazino)-ethanamine, a nitric oxide donor. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-kinase), did not block the protective effect of vanadate, suggesting that the protective effect of vanadate is independent on PI3-kinase. Vanadate increased tyrosine phosphorylation of several proteins including the focal adhesion protein p130 Crk-associated substrate (p130(cas)). By the treatment with SIN-1, the endogenous association of p130(cas) and Crk was disrupted, and the association was restored by vanadate treatment. These results suggest that disruption of tyrosine phosphorylation signaling may be critical for peroxynitrite-induced cell death, and that vanadate prevents cell death at least in part through the enhancement in tyrosine phosphorylation of the proteins including p130(cas).


Retroinfusion of NFkappaB decoy oligonucleotide extends cardioprotection achieved by CD18 inhibition in a preclinical study of myocardial ischemia and retroinfusion in pigs.
Myocardial reperfusion injury is partially mediated by postischemic inflammation. Beyond acute PMN recruitment, postischemic inflammation comprises subacute PMN adhesion, eg via NFkappaB activation. In a pig model of 60-min LAD occlusion by PTCA ballon inflation and 1 to 7 days of reperfusion, we investigated the impact of targeted NFkappaB decoy oligonucleotide (ODN) transfection in the area at risk (AAR) on infarct size and regional myocardial function. After 55 min of LAD occlusion, liposomes containing NFkappaB ODN were selectively retroinfused into the anterior interventricular vein for 5 min. Then, retroinfusion was stopped and reperfusion was initiated. Where indicated, CD18 antibody IB4 was infused systemically at 30 min of ischemia. Methylen blue and tetrazolium-red staining were used for quantification of the infarct size. Subendocardial segment shortening (SES) by sonomicrometric crystals in infarct area and AAR was assessed under pacing (expressed as % of control region). NFkappaB decoy ODN retroinfusion reduced infarct size (36 +/- 4% versus 49 +/- 5% in control hearts at day 7), whereas functional reserve of the AAR (SES 73 +/- 17% versus 46 +/- 18% at 180/min) tended to improve. Similar effects were observed after IB4 infusion (38 +/- 5% infarct size, 85 +/- 7% SES at 180/min). A combination of NFkappaB decoy ODN retroinfusion and IB4 infusion further decreased infarct size (26 +/- 2%) and improved functional reserve (SES 94 +/- 6% at 180/min). We conclude that NFkappaB decoy ODN transfection by retroinfusion is feasible in pig hearts and provides postischemic cardioprotection in addition to CD18 blockade.


Bronchoscopy in severe childhood asthma: Irresponsible or irreplaceable?
For children with severe asthma, guideline-based management focuses on the escalation of anti-inflammatory and bronchodilatory medications while addressing comorbid conditions. Bronchoscopy, in this context, has been relegated to ruling out asthma mimickers. More recently, however, there have been questions surrounding the clinical utility of bronchoscopy in severe childhood asthma. In this solicited lecture summary, we discuss the past, present, and potential future applications of bronchoscopy in severe childhood asthma.


HCC complicated by PVT: Outcome and the role of anticoagulation therapy.
This retrospective study investigated the clinical impact of PVT on the course of patients with HCC who were and were not treated with anticoagulation (AC). We retrospectively evaluated a cohort of 60 patients diagnosed with HCC and PVT. Nine patients were excluded for lack of follow-up. HCC, PVT diagnosis, and imaging follow-up were performed using contrast-enhanced computed tomography or MRI. Of the 51 patients evaluated, 12 received AC and 39 did not. Forty-two patients were male; mean age was 60.3 years. Mean survival after HCC diagnosis was 32.9 months; after PVT diagnosis, it was 18.4 months. No symptoms directly related to PVT development were reported. AC therapy was initiated for 12 patients and had to be discontinued for 3 patients because of complications. AC was not associated with a difference in PVT progression (49% in non-AC group vs. 50% in AC group). After adjusting for age, HCC type (single vs. multifocal), and Child-Pugh score, AC was associated with an improved survival after HCC diagnosis (adjusted hazard ratio [HR] = 0.37; 95% confidence interval [CI] 0.14 to 0.99) and after PVT diagnosis (HR = 0.34; 95% CI 0.13 to 0.88). Patients with HCC complicated by PVT in both AC and non-AC groups had a similar rate of progression. Neither group had symptoms attributable to PVT. Possible AC-related complications need to be considered before proceeding with therapy in patients with HCC and PVT. AC may be associated with a survival advantage in patients with HCC and PVT.


Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during allergic airway disease.
The NADPH oxidase DUOX1 contributes to epithelial production of alarmins, including interleukin (IL)-33, in response to injurious triggers such as airborne protease allergens, and mediates development of mucus metaplasia and airway remodeling in chronic allergic airways diseases. DUOX1 is also expressed in non-epithelial lung cell types, including macrophages that play an important role in airway remodeling during chronic lung disease. We therefore conditionally deleted DUOX1 in either lung epithelial or monocyte/macrophage lineages to address its cell-specific actions in innate airway responses to acute airway challenge with house dust mite (HDM) allergen, and in chronic HDM-driven allergic airway inflammation. As expected, acute responses to airway challenge with HDM, as well as type 2 inflammation and related features of airway remodeling during chronic HDM-induced allergic inflammation, were largely driven by DUOX1 with the respiratory epithelium. However, in the context of chronic HDM-driven inflammation, DUOX1 deletion in macrophages also significantly impaired type 2 cytokine production and indices of mucus metaplasia. Further studies revealed a contribution of macrophage-intrinsic DUOX1 in macrophage recruitment upon chronic HDM challenge, as well as features of macrophage activation that impact on type 2 inflammation and remodeling.


Case Report of Conjunctival Melanoacanthoma.
The purpose of this study was to describe a case of conjunctival melanoacanthoma, an exceedingly rare condition that has yet to be fully described in the literature. Melanoacanthomas are most commonly seen on the skin or oral mucosa and are believed to result from local irritation or trauma. A 34-year-old Hispanic man presented with a painless, solitary, pigmented conjunctival lesion, in addition to bilateral pterygia suggesting chronic solar damage. The lesion was excised and sent for analysis. Histopathologic analysis of tissue samples demonstrated melanocyte proliferation and epithelial dysplasia, yielding a final pathologic diagnosis of conjunctival melanoacanthoma with dysplastic and acantholytic-type features. The patient is being closely followed and has not had recurrence of the lesion. Only 1 prior case of conjunctival melanoacanthoma has been documented. As such, there is no standard of care regarding appropriate management.


Detection of the SARS-CoV-2 spike protein in saliva with Shrinky-Dink© electrodes.
Using the children's toy, Shrinky-Dink ©, we present an aptamer-based electrochemical (E-AB) assay that recognizes the spike protein of SARS-CoV-2 in saliva for viral infection detection. The low-cost electrodes are implementable at population scale and demonstrate detection down to 0.1 fg mL -1 of the S1 subunit of the spike protein.


ANO5 mutations in the Dutch limb girdle muscular dystrophy population.
A Dutch cohort of 105 limb girdle muscular dystrophy (LGMD) patients were subject to subsequent genetic investigations. In half the families a causative mutation was found. Recently mutations were identified in ANO5 causing LGMD2L and Miyoshi-like myopathy (MMD3), but could also be found in patients with hyperCKemia only. Therefore, we analysed the index cases of the remaining 31 as yet undiagnosed families from our previously described cohort of LGMD patients for the presence of ANO5 mutations. Detailed history and neurological examination were available for all patients. Serum creatine kinase (CK) activity, skeletal muscle computed tomography (CT) and cardiological investigations were performed. Mutations in ANO5 were found in 16% of the families: 11 index patients and two sibs, eight males and five females. The founder mutation c.191dupA was present in 8 out of 13 patients. Ten different pathogenic mutations were identified of which seven were novel: five missense and two splice site mutations. The age of these patients ranged from 26 to 69 years and the age of onset varied from 21 to 57 years. Symptoms at onset were related to proximal leg weakness. The weakness was slowly progressive. Calf hypertrophy was present in three patients. Males were more severely affected than females. Serum CK activity was highly elevated in the early stage of disease and moderately increased in later stages. Muscle biopsy showed predominantly dystrophic changes. One patient had hypertrophic cardiomyopathy, two others had intraventricular septum thickening.


Glycolysis inhibition ameliorates brain injury after ischemic stroke by promoting the function of myeloid-derived suppressor cells.
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells which are immunosuppressive and glycolytically inactive in inflammatory diseases. However, it is unknown whether MDSCs contribute to ischemic stroke and how glycolysis regulates MDSC function in such a context. Here, we showed that MDSCs arise in the blood of patients at early phase of stroke. Similar results were observed in temporary middle cerebral artery occlusion-induced cerebral ischemic mice. Pharmaceutical exhaustion of MDSCs aggravated, while adoptive transfer of MDSCs rescued the ischemic brain injury. However, the differentiation of MDSCs into immunopotent myeloid cells which coincides with increased glycolysis was observed in the context of ischemic stroke. Mechanistically, the glycolytic product lactate autonomously induces MDSC differentiation through activation of mTORC1, and paracrinely activates Th1 and Th17 cells. Moreover, gene knockout or inhibition of the glycolytic enzyme PFKFB3 increased endogenous MDSCs by blocking their differentiation, and improved ischemic brain injury. Collectively, these results revealed that glycolytic switch decreases the immunosuppressive and neuroprotective role of MDSCs in ischemic stroke and pharmacological targeting MDSCs via glycolysis inhibition constitutes a promising therapeutic strategy for ischemic stroke.


Disabling chronic low back pain with Modic type 1 MRI signal: acute reduction in pain with intradiscal corticotherapy.
The objective of the current study was to compare short- and long-term effect on chronic low back pain of intradiscal injection of methylprednisolone with or without presence of Modic type 1 MRI changes. Medical charts of patients receiving intradiscal injection of methylprednisolone from January 1, 1995 to December 31, 1998 were retrospectively reviewed. Clinical parameters were recorded at baseline, 24h after injection and at follow-up (12-14 months). Patients were studied in three groups: Modic I-a, if patients had Modic type 1 changes with no previous surgery or nucleolysis (n=30); Modic I-b, if patients had Modic type 1 changes at the level of previous surgery or nucleolysis (n=37); Control, if patients had no Modic type 1 changes (n=30). Twenty-four hours after methylprednisolone injection, higher proportion of patients with self-assessed improvement was observed in Modic I-a (90%) and Modic I-b (71%) than in Control (30%). Low back pain decreased in both Modic groups. Low back pain did not vary from baseline in controls. No effect was detected in three groups, neither for radiating pain 24h after injection, nor for any outcome parameters at the latest follow-up. We suggest that patients with disabling chronic low back pain and Modic type 1 MRI changes have specific acute response to intradiscal injection of methylprednisolone. Clinical studies are however necessary to further investigate the effectiveness and safety of such injections.


Analgesic efficacy of piroxicam in postoperative dental pain.
The severity of postoperative dental pain can be variable depending on the type of procedure. Both centrally acting and peripherally acting analgesics, such as nonsteroidal anti-inflammatory drugs (NSAIDs), aspirin, and acetaminophen are used. NSAIDs are generally better suited to ambulatory outpatients. The most commonly used postoperative dental pain model includes patients who have undergone surgical removal of impacted third molar teeth. The analgesic efficacy of piroxicam in this pain model was studied both in the United States and in foreign centers. The foreign studies suggest that piroxicam at 20-mg doses produces analgesia in patients with postoperative dental pain. Seven single-dose, randomized, double-blind trials of 798 patients in the United States more clearly evaluated the efficacy of piroxicam. These studies used various doses of piroxicam (5, 10, 20, and 40 mg), aspirin 648 mg, and placebo. Safety results showed that a wide range of piroxicam doses were safe when administered in single doses. Although neither piroxicam 5 mg nor 10 mg produced clinically significant analgesia, 20-mg and 40-mg doses were significantly superior to placebo and both were comparable with aspirin 648 mg over the initial six hours. Piroxicam 20 mg and 40 mg, however, produced significantly longer durations of analgesia than aspirin 648 mg, and it appears that the analgesic effect of piroxicam may extend for up to 24 hours in a substantial proportion of patients.


Weekend effect on 30-day mortality for ischemic and hemorrhagic stroke analyzed using severity index and staffing level.
Previous studies on the weekend effect-a phenomenon where stroke outcomes differ depending on whether the stroke occurred on a weekend-mostly targeted ischemic stroke and showed inconsistent results. Thus, we investigated the weekend effect on 30-day mortality in patients with ischemic or hemorrhagic stroke considering the confounding effect of stroke severity and staffing level. We retrospectively analyzed data of patients hospitalized for ischemic or hemorrhagic stroke between January 1, 2015, and December 31, 2018, which were extracted from the claims database of the National Health Insurance System and the Medical Resource Report by the Health Insurance Review & Assessment Service. The primary outcome measure was 30-day all-cause mortality. In total, 278,632 patients were included, among whom 84,240 and 194,392 had a hemorrhagic and ischemic stroke, respectively, with 25.8% and 25.1% of patients, respectively, being hospitalized during the weekend. Patients admitted on weekends had significantly higher 30-day mortality rates (hemorrhagic stroke 16.84%>15.55%, p<0.0001; ischemic stroke 5.06%>4.92%, p<0.0001). However, in the multi-level logistic regression analysis adjusted for case-mix, pre-hospital, and hospital level factors, the weekend effect remained consistent in patients with hemorrhagic stroke (odds ratio [OR] 1.05, 95% confidence interval [CI] 1.00-1.10), while the association was no longer evident in patients with ischemic stroke (OR 1.01, 95% CI 0.96-1.06). Weekend admission for hemorrhagic stroke was significantly associated with a higher mortality rate after adjusting for confounding factors. Further studies are required to understand factors contributing to mortality during weekend admission.


Synthesis, reactivity, and biological activity of 5-aminouracil and its derivatives.
The synthesis and reactions of 5-aminouracil and its derivatives are comprehensively reviewed. A brief survey of biological activities, especially chemotherapeutic and pharmacological properties, is also reported.


Rapid and specific SPRi detection of L. pneumophila in complex environmental water samples.
Legionellosis is a very devastating disease worldwide mainly due to unpredictable outbreaks in man-made water systems. Developing a highly specific and sensitive rapid detection system that detects only metabolically active bacteria is a main priority for water quality assessment. We previously developed a versatile technique for sensitive and specific detection of synthetic RNA. In the present work, we further investigated the performance of the developed biosensor for detection of Legionella pneumophila in complex environmental samples, particularly those containing protozoa. The specificity and sensitivity of the detection system were verified using total RNA extracted from L. pneumophila in spiked water co-cultured with amoebae. We demonstrated that the expression level of ribosomal RNA (rRNA) is extremely dependent on the environmental conditions. The presence of amoebae with L. pneumophila, especially in nutrition-deprived samples, increased the amount of L. pneumophila 15-fold after 1 week as measured through the expression of 16s rRNA. Using the developed surface plasmon resonance imaging (SPRi) detection method, we were also able to successfully detect L. pneumophila within 3 h, both in the presence and absence of amoebae in the complex environmental samples obtained from a cooling water tower. These findings suggest that the developed biosensing system is a viable method for rapid, real-time and effective detection not only for L. pneumophila in environmental samples but also to assess the risk associated with the use of water contaminated with other pathogens.


Exudative retinopathy and detachment: a late reactivation of retinopathy of prematurity after intravitreal bevacizumab.
A 25-week postmenstrual age premature infant was treated with bilateral intravitreal bevacizumab for retinopathy of prematurity at 35 weeks' PMA. Postinjection, the retinal vessels progressed anteriorly within the retina. The patient presented 1 year after injection with bilateral exudative retinal detachments. The right eye was treated with intravitreal bevacizumab, laser ablation, and scleral buckling, resulting in resolution of the exudation and detachment. The left eye was treated with vitrectomy and lensectomy, but persistent exudation and detachment remained. This case demonstrates the rare complication of exudative retinal detachment in the setting of retinopathy of prematurity, which may become more common with increasing use of bevacizumab. Importantly, it also demonstrates the need not only for frequent examination after bevacizumab injection for retinopathy of prematurity but long-term follow-up as well, until either the retina is fully vascularized or peripheral ablation is performed.


Brain metabolic and microstructural alterations associated with hepatitis C virus infection, autoimmune hepatitis and primary biliary cholangitis.
Neuropsychiatric symptoms in hepatitis C (HCV) patients resemble those of patients with autoimmune hepatitis (AIH) or primary biliary cholangitis (PBC), whilst the mechanisms behind them are unknown. Here we looked for cerebral metabolic and/or microstructural alterations in patients with HCV, AIH or PBC as possible causes behind these symptoms. Patients with HCV infection (n = 17), AIH (n = 14) or PBC (n = 11) and age-adjusted healthy controls (n = 18) underwent brain magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS) and psychometric assessment of memory and attention. Brain relative proton density (PD) and T2 relaxation time (T2) were determined in 17 regions of interest (ROIs), as were the concentrations of N-acetyl-aspartate, choline, creatine, myo-inositol and glutamine + glutamate in frontal- (fWM) and parietal white matter (pWM). One-way analysis of variance and Kruskal-Wallis tests were used for group comparison. Correlations between altered neuropsychological findings and MRI/MRS observations were estimated with the Spearman ρ test. HCV, AIH and PBC patients revealed similar alterations in brain PD and metabolites compared to controls: significantly decreased PD in 7/17 ROIs in the HCV group, 16/17 ROIs in the PBC group and 14/17 ROIs in the AIH group, significantly increased N-acetyl-aspartate in fWM in all patients, significantly increased choline in the PBC group in both fWM and pWM, in the AIH group only in pWM and with a trend in the HCV group in pWM. Correlation analysis did not reveal significant associations between MRI/MRS alterations and neuropsychological dysfunction. The findings suggest similar pathophysiological mechanisms behind neuropsychiatric symptoms associated with HCV infection, AIH and PBC.


Systemic membrane defect and the inhibition of lymphocyte capping in Duchenne muscular dystrophy.
Eight reversible inhibitors were used to study decreases in lymphocyte capping in patients with Duchenne Muscular Dystrophy (DMD) when compared to controls. The inhibitors included hydrocortisone, chlorpromazine, calcium ionophore, Cytochalasin D, propranolol, dibucaine, fluoride and azide. All of these inhibitors disrupt cap formation. Mononuclear leukocytes from DMD patients and controls were isolated from whole blood, incubated with fluorescein-conjugated polyvalent antisera and inhibitor, induced to form caps, and the caps counted using a fluorescent microscope. Cell viabilities and morphology were assessed. After removal of inhibitor, the cells were recounted. All of the inhibitors significantly lowered capping in controls (p less than 0.001), but this effect was seen with only four out of the eight inhibitors in DMD patients. Dibucaine and azide were less inhibitory in patients (p less than 0.005, p greater than 0.05, and p greater than 0.05 respectively) while capping in patients was not inhibited by fluoride and hydrocortisone (p greater than 0.5). The lack of hydrocortisone inhibition suggests that the differences in capping between DMD patients and controls may lie within the membrane itself, rather than its associated components (i.e. cytoskeletal network), and that the defect occurs toward the beginning of the capping sequence.


United Kingdom National Ophthalmology Database study of vitreoretinal surgery: report 3, retinal detachment.
To describe rhegmatogenous retinal detachment (RD) surgery. National Ophthalmology Database study. A total of 3403 eyes from 3321 patients undergoing primary RD surgery. Participating centers prospectively collected clinical data using a single electronic medical record system, with automatic extraction of anonymized data to a national database, from 2002 to 2010. Description of the primary procedures performed, intraoperative complication rate, and proportion of eyes undergoing subsequent RD or cataract surgery. We undertook an exploratory analysis of change in visual acuity (VA) using the data available. Of 3403 operations, 2693 (79.1%) were pars plana vitrectomy (PPV), 413 (12.1%) were retinopexy with a scleral buckle (SB), and 297 (8.7%) were PPV with an SB (PPV-SB). For PPV and PPV-SB, 18.8% were with hexafluoroethane, 12.1% were with perfluoropropane, 43.1% were with sulfahexafluoride, 1.8% were with air, 17.9% were with silicone oil, and 10.7% were with cataract surgery. Within 1 year of vitrectomy, 52.1% of phakic eyes had undergone cataract surgery. For all RD operations combined (and excluding cataract surgery complications), 5.1% had 1 or more intraoperative complication, 13.0% underwent further RD surgery, and 8.3% had silicone oil in situ at last review. The RD reoperation rate was 13.3%, 12.3%, and 14.5% for PPV, SB, and PPV-SB, respectively. For 961 eyes with a baseline and final VA measurement, the median presenting logarithm of the minimum angle of resolution VA improved from 1.0 to 0.5 (20/200-20/63) after a median follow-up of 0.6 years. These results may help vitreoretinal surgeons to benchmark their intraoperative complication rate and reoperation rate and to compare their surgical techniques with their peers'. They suggest that the benefits of RD surgery greatly outweigh the risks.


Influence of river discharge on abundance and dissemination of heterotrophic, indicator and pathogenic bacteria along the East Coast of India.
In order to examine the influence of discharge from different rivers from peninsular India and urban sewage on intensity and dissemination of heterotrophic, indicator and pathogenic bacteria, a study was carried out during peak discharge period along coastal Bay of Bengal. The coastal Bay received freshwater inputs from the river Ganges while Godavari and Krishna contributed to the south. Contrasting difference in salinity, temperature, nutrients and organic matter was observed between north and south east coast of India. The highest heterotrophic, indicator and pathogenic bacterial abundance was observed in the central coastal Bay that received urban sewage from the major city. Intensity and dissemination of heterotrophic, indicator and pathogenic bacteria displayed linear relation with magnitude of discharge. The coliform load was observed up to 100km from the coast suggesting that marine waters were polluted during the monsoon season and its impact on the ecosystem needs further studies.


Early salvage radiotherapy following radical prostatectomy.
Depending on the pathologic tumour stage, up to 60% of prostate cancer patients who undergo radical prostatectomy will develop biochemical relapse and require further local treatment. We reviewed the results of early salvage radiation therapy (RT), defined as prostate-specific antigen (PSA) values prior to RT ≤ 0.5 ng/ml in the setting of lymph node-negative disease. Ten retrospective studies, including one multicentre analysis, were used for this analysis. Among them, we received previously unpublished patient characteristics and updated outcome data from five retrospective single-centre trials to perform a subgroup analysis for early salvage RT. Patients treated with early salvage RT have a significantly improved biochemical recurrence-free survival (BRFS) rate compared with those receiving salvage RT initiated after PSA values are >0.5 ng/ml. Similarly, within the cohort of patients with pre-RT PSA values <0.5 ng/ml, improved BRFS rates were noted among those with lower rather higher pre-RT PSA levels. It is possible that higher RT dose levels and the use of adjunctive androgen-deprivation therapy improve biochemical control outcomes in the salvage setting. Based on a literature review, improved 5-yr BRFS rates are observed for patients who receive early salvage RT compared with patients treated with salvage RT with a pre-RT PSA value >0.5 ng/ml. Whether the routine application of early salvage RT in patients with initially undetectable PSA levels will be associated with demonstrable clinical benefit awaits the results of ongoing prospective trials.


Kind and distribution of cutaneous sensation loss in hereditary transthyretin amyloidosis with polyneuropathy.
Report on the kind and distribution of somatotopic sensation loss and its utility in assessing severity of sensation loss in study of a large international cohort of patients with hereditary transthyretin amyloidosis with polyneuropathy (hATTR-PN). Smart Somatotopic Quantitative Sensation Testing (S ST QSTing) using Computer Assisted Sensation Evaluator IVc (CASE IVc) was used to assess the somatotopic distribution of touch pressure (TP) and heat pain (HP) sensation loss twice of untreated hATTR-PN patients in the Ionis NEURO-TTR trial (www.clinicaltrials.gov, NCT01737398). Of the studied cohort of 169 patients, 163 (97%) had sensation loss, both TP and HP in 121/169 (75%), TP only in 39/169 (23%), and HP only in 3/169 (2%). Sensation loss typically affected both lower (152/169-90%) and upper limb (135/169-82%), and overall TP sensation loss was greater than HP loss, except for early-onset Val30Met patients in which HP exceeded TP loss. Using S ST QSTing, a highly quantitated, standardized, referenced, and automated QSTing approach of the body's surface distribution of sensation loss we have shown that: 1) reliable and useful measurement of the body surface distribution of sensation loss is possible; 2) this measure is abnormal in most patients with hATTR-PN and is an indication of polyneuropathy severity; and 3) cutaneous sensation loss involves both large and small sensory fibers in this disease but slightly more small fibers in early onset Val30Met patients.


Integrin alphaIIb-subunit cytoplasmic domain mutations demonstrate a requirement for tyrosine phosphorylation of beta3-subunits in actin cytoskeletal organization.
Using truncated or mutated alphaIIb integrin cytoplasmic domains fused to the alphaV extracellular domain and expressed with the beta3 integrin subunit, we demonstrate that the double mutation of proline residues 998 and 999 to alanine (PP998/999AA), previously shown to disturb the C-terminal conformation of the alphaIIb integrin cytoplasmic domain, prevents tyrosine phosphorylation of beta3 integrin induced by Arg-Gly-Asp peptide ligation. This mutation also inhibits integrin mediated actin assembly and cell adhesion to vitronectin. In contrast, progressive truncation of the alphaIIb-subunit cytoplasmic domain did not reproduce these effects. Interestingly, the PP998/999AA mutations of alphaIIb did not affect beta3 tyrosine phosphorylation, cell adhesion, or actin polymerization induced by manganese. Exogenous addition of manganese was sufficient to rescue beta3 phosphorylation, cell adhesion, and actin assembly in cells expressing the PP998/999AA mutation when presented with a vitronectin substrate. Further, induction of the high affinity conformation of this mutant beta3 integrin by incubation with either Arg-Gly-Asp peptide or exogenous manganese was equivalent. These results suggest that the extracellular structure of beta3 integrins in the high affinity conformation is not directly related to the structure of the cytoplasmic face of the integrin. Moreover, the requirement for beta3 phosphorylation is demonstrated without mutation of the beta3 subunit. In support of our previous hypothesis of a role for beta3 phosphorylation in adhesion, these studies demonstrate a strong correlation between beta3 tyrosine phosphorylation and assembly of a cytoskeleton competent to support firm cell adhesion.


New insights into the regulation of myosin light chain phosphorylation in retinal pigment epithelial cells.
The retinal pigment epithelium (RPE) plays an essential role in the function of the neural retina and the maintenance of vision. Most of the functions displayed by RPE require a dynamic organization of the acto-myosin cytoskeleton. Myosin II, a main cytoskeletal component in muscle and non-muscle cells, is directly involved in force generation required for organelle movement, selective molecule transport within cell compartments, exocytosis, endocytosis, phagocytosis, and cell division, among others. Contractile processes are triggered by the phosphorylation of myosin II light chains (MLCs), which promotes actin-myosin interaction and the assembly of contractile fibers. Considerable evidence indicates that non-muscle myosin II activation is critically involved in various pathological states, increasing the interest in studying the signaling pathways controlling MLC phosphorylation. Particularly, recent findings suggest a role for non-muscle myosin II-induced contraction in RPE cell transformation involved in the establishment of numerous retinal diseases. This review summarizes the current knowledge regarding myosin function in RPE cells, as well as the signaling networks leading to MLC phosphorylation under pathological conditions. Understanding the molecular mechanisms underlying RPE dysfunction would improve the development of new therapies for the treatment or prevention of different ocular disorders leading to blindness.


A water resistant solid-phase microextraction fiber with high selectivity prepared by a metal organic framework with perfluorinated pores.
A novel solid-phase microextraction (SPME) fiber was fabricated by the construction of fluorous metal organic frameworks (FMOF) and a polyimide (PI) composite strategy. As an auxiliary material, PI was expected to help FMOF particles form well-knit film on the surface of stainless steel wire and reinforce the coating, and FMOF was expected to afford a special structure to absorb, extract and enrich. Furthermore, it was explored for the headspace SPME (HS-SPME) of six volatile aromatic compounds (VACs) from water samples followed by gas chromatographic (GC) separation with flame ionization detection. Under the optimized conditions, the fiber afforded wide linear ranges (1-1000 μgL(-1)), low detection limits (0.15-0.9 μgL(-1)) and acceptable repeatability (<4.6%) and reproducibility (<7.3%). The FMOF@PI coated fiber not only offered large enhancement factors for benzene (1227) but also exhibited high extraction selectivity for benzene to other benzene homologues, hydrocarbons and phenols; for example, the extraction ratio of benzene to toluene, n-hexane and phenol could be as high as 10.2, 64.1 and 32.3, respectively. Moreover, the FMOF@PI-coated fiber afforded good thermal, water and organic solvent stabilities, and a long lifetime (over 200 times). The developed method was successfully applied to the determination of VACs in wastewater samples.


Pretreatment growth rate predicts radiation response in vestibular schwannomas.
Vestibular schwannomas (VS) are often followed without initial therapeutic intervention because many tumors do not grow and radiation therapy is associated with potential adverse effects. In an effort to determine whether maximizing initial surveillance predicts for later treatment response, the predictive value of preirradiation growth rate of VS on response to radiation therapy was assessed. Sixty-four patients with 65 VS were treated with single-fraction stereotactic radiation surgery or fractionated stereotactic radiation therapy. Pre- and postirradiation linear expansion rates were estimated using volumetric measurements on sequential magnetic resonance images (MRIs). In addition, postirradiation tumor volume change was classified as demonstrating shrinkage (ratio of volume on last follow-up MRI to MRI immediately preceding irradiation <80%), stability (ratio 80%-120%), or expansion (ratio >120%). The median pre- and postirradiation follow-up was 20.0 and 27.5 months, respectively. Seven tumors from neurofibromatosis type 2 (NF2) patients were excluded from statistical analyses. In the 58 non-NF2 patients, there was a trend of correlation between pre- and postirradiation volume change rates (slope on linear regression, 0.29; P=.06). Tumors demonstrating postirradiation expansion had a median preirradiation growth rate of 89%/year, and those without postirradiation expansion had a median preirradiation growth rate of 41%/year (P=.02). As the preirradiation growth rate increased, the probability of postirradiation expansion also increased. Overall, 24.1% of tumors were stable, 53.4% experienced shrinkage, and 22.5% experienced expansion. Predictors of no postirradiation tumor expansion included no prior surgery (P=.01) and slower tumor growth rate (P=.02). The control of tumors in NF2 patients was only 43%. Radiation therapy is an effective treatment for VS, but tumors that grow quickly preirradiation may be more likely to increase in size. Clinicians should take into account tumor growth rate when counseling patients about treatment options.


Timely identification and isolation of affected individuals is one of the most important factors to control spreading of the newly emerged SARS-CoV-2. Consequently, it is crucial to maintain sufficient sample capacities in diagnostic laboratories. Here, we present a high-through-put approach for PCR-based SARS-CoV-2 testing. The implementation of sample pooling reduces costs and workload, especially in times with low population prevalence.


Synthesis and in vivo evaluation of (E)-N-[(11)C]Methyl-4- (3-pyridinyl)-3-butene-1-amine ([(11)C]metanicotine) as a nicotinic receptor radioligand.
(E)-N-[(11)C]Methyl-4-(3-pyridinyl)-3-butene-1-amine ([(11)C]metanicotine), a high affinity (K(i) = 16 nM) CNS-selective nicotinic agonist, was prepared by the [(11)C]alkylation of the desmethyl precursor with [(11)C]methyl trifluoromethanesulfonate. In vivo distribution studies in mice demonstrated good blood brain permeability but essentially uniform regional brain distribution and no evidence of specific binding to nicotinic cholinergic receptors. Identical results were obtained in an imaging study performed in a monkey brain. Therefore, despite literature reports supporting the use of metanicotine as a cognition enhancing nicotinic agonist, (E)-N-[(11)C]methyl-4-(3-pyridinyl)-3-butene-1-amine does not appear to be a suitable candidate for in vivo imaging studies of nicotinic acetylcholine receptors in the mammalian brain.


Serum hyaluronic acid for the assessment of graft viability in porcine liver transplantation.
For the assessment of graft viability, serum hyaluronic acid (HA) levels during porcine orthotopic liver transplantation were measured in two groups: group 1 (viable: n = 5) in which allografts were transplanted following a minimal cold (4 degrees C) preservation, and group 2 (nonviable: n = 4) in which allografts were transplanted after cold static storage (4 degrees C) for 24 h in University of Wisconsin solution. The changes in the HA levels reached a significant difference between the two groups at 30 min after reperfusion (P < 0.02). In group 1, all animals survived for over 4 days, while all animals in group 2 died within 24 h. The serum HA also demonstrated a significant correlation with prothrombin time, beta-glucuronidase, and aspartate aminotransferase at 120 min after reperfusion. These results suggest that the measurement of serum HA is a potentially effective index for evaluating hepatic allograft viability.