Abnormal expression of cerebrospinal fluid cation chloride cotransporters in patients with Rett syndrome.
Rett Syndrome is a progressive neurodevelopmental disorder caused mainly by mutations in the gene encoding methyl-CpG-binding protein 2. The relevance of MeCP2 for GABAergic function was previously documented in animal models. In these models, animals show deficits in brain-derived neurotrophic factor, which is thought to contribute to the pathogenesis of this disease. Neuronal Cation Chloride Cotransporters (CCCs) play a key role in GABAergic neuronal maturation, and brain-derived neurotrophic factor is implicated in the regulation of CCCs expression during development. Our aim was to analyse the expression of two relevant CCCs, NKCC1 and KCC2, in the cerebrospinal fluid of Rett syndrome patients and compare it with a normal control group. The presence of bumetanide sensitive NKCC1 and KCC2 was analysed in cerebrospinal fluid samples from a control pediatric population (1 day to 14 years of life) and from Rett syndrome patients (2 to 19 years of life), by immunoblot analysis. Both proteins were detected in the cerebrospinal fluid and their levels are higher in the early postnatal period. However, Rett syndrome patients showed significantly reduced levels of KCC2 and KCC2/NKCC1 ratio when compared to the control group. Reduced KCC2/NKCC1 ratio in the cerebrospinal fluid of Rett Syndrome patients suggests a disturbed process of GABAergic neuronal maturation and open up a new therapeutic perspective.


Preoperative statin administration does not protect against early postoperative acute respiratory distress syndrome: a retrospective cohort study.
Statins have been shown to possess antiinflammatory and immunomodulatory effects. In this study, we sought to determine if preoperative statin therapy is associated with a reduced frequency of postoperative acute respiratory distress syndrome (ARDS) in surgical populations at increased risk of developing ARDS. We performed a retrospective cohort evaluation of the association between preoperative statin therapy and early postoperative ARDS in patients undergoing elective high-risk thoracic and aortic vascular surgery. The association between preoperative statin therapy and postoperative ARDS was assessed using propensity-adjusted analyses to control for indication bias and confounding factors. Of 1845 patients, 722 were receiving preoperative statin therapy. One hundred twenty patients developed postoperative ARDS. Frequencies of ARDS among those receiving statin therapy versus those who were not was 7.2% and 6.1%, respectively (OR = 1.20; 95% CI, 0.83-1.75; P = 0.330). Neither the stratified propensity score analysis (pooled OR 0.93; 95% CI, 0.60-1.43) nor matched analysis (OR = 0.78; 95% CI, 0.48-1.27) identified a statistically significant association between preoperative statin administration and postoperative ARDS. When compared to matched controls, patients who developed postoperative ARDS did not differ in mortality (7.7% vs 8.8%, P = 0.51), hospital length of stay (21 days vs 15 days, P = 0.21), or ventilator-free days (24 days vs 25 days, P = 0.62). In patients undergoing high-risk surgery, preoperative statin therapy was not associated with a statistically significant reduction in postoperative ARDS. These results do not support the use of statins as prophylaxis against ARDS in patients undergoing high-risk surgery.


Quantifying the information content of lattice images.
Quantitative information may be extracted from local areas of images that consist of one or more types of unit cell. Fourier-space analysis, real-space intensity analysis, and real-space vector pattern recognition are discussed. The pattern recognition approach efficiently exploits the available information by representing the intensity distribution within each unit cell of the image as a multidimensional vector. Thus, the amount and the effect of noise present are determined, statistically significant features are identified, and quantitative comparisons are made with model images. In the case of chemical lattice images, the position of a vector can be directly related to the atomic composition of the unit cell it represents, allowing quantitative chemical mapping of materials at near-atomic sensitivity and resolution. More generally, the vector approach allows the efficient and quantitative extraction of information from images, which consist of mosaics of unit cells.


New antibiotics for healthcare-associated pneumonia.
Current antibiotics available for the treatment of healthcare-associated pneumonia (HCAP) may result in clinical failure due to resistance development, side effect intolerance, or poor pharmacokinetic-pharmacodynamic profiles. New agents active against common HCAP pathogens are needed. The mechanism of action, spectrum of activity, pharmacokinetics, adverse effects, and clinical efficacy of seven new agents in clinical development or recently approved with either methicillin-resistant Staphylococcus aureus (MRSA) or pseudomonal activity are reviewed. They include doripenem, a new antipseudomonal carbapenem; ceftobiprole and ceftaroline, two anti-MRSA cephalosporins; iclaprim, a selective dihydrofolate reductase antagonist; and three glycopeptides, dalbavancin, telavancin, and oritavancin.


Cluster headache--course over ten years in 189 patients.
One-hundred-and-eighty-nine cluster headache patients, referred to Parma and Pavia Headache Centres between 1976 and 1986 with a disease duration of over 10 years, were interviewed about the course of cluster headache. They were classified as episodic (n = 140) or chronic (n = 49) cluster headache patients on the basis of course during the year of onset. Episodic patients showed the following outcome: maintenance of an episodic form (primary episodic form) in 80.7% of cases, shift towards a chronic form (secondary chronic form) in 12.9% and shift towards an intermediate pattern ("combined" form) in 6.4%. In chronic patients, cluster headache was still chronic (primary chronic form) at the moment of observation in 52.4% of cases, while it turned into an episodic form ("secondary" episodic form) in 32.6% and into a "combined" form in 14.3%. Nineteen patients (10%) had had no attacks for at least three years at the moment of examination. We can conclude from our data that: cluster headache is a disease of long duration, perhaps lifelong; episodic cluster headache tends to worsen; chronic cluster headache may easily turn into a better prognostic episodic form; prophylactic drugs are unable to induce recovery. The following factors seem related to a poor outcome: a later onset, the male gender and a disease duration of over 20 years for the episodic forms.


Maternal mortality in a transitional hospital in Enugu, south east Nigeria.
Nigeria has one of the highest maternal mortality ratios in the world. The study was to determine the trend of maternal mortality ratio in the hospital as it transits from a General through a Specialist to a Teaching hospital. It was a retrospective review of maternal deaths at Enugu State University Teaching Hospital Parklane, over its 5 year transition period (January 2004 to December 2008). There were 7146 live births and 60 maternal deaths giving an overall maternal mortality ratio (MMR) of 840/100,000 livebirths. The MMR rose from 411 to 1137/100,000 live births as a specialist hospital, with a decline to 625/100,000 as a Teaching hospital. Pre-eclampsia/eclampsia was the leading cause (29.63%) of maternal death. MMR was highest as a Specialist hospital due to limited manpower and inadequate facilities to properly manage the rising number of referred obstetric emergencies. Adequate preparations should be made before upgrading a hospital, to enable it cope with the challenges of managing referred obstetric emergencies


Recent advances in neoadjuvant chemoradiotherapy in locally advanced rectal cancer.
Preoperative chemoradiotherapy (CRT) has been actively used in Europe and the United States to treat advanced low rectal cancer, and provides excellent local control. In Japan, however, the standard treatment is lateral lymph node dissection, and to date CRT has not been actively used. In recent years, an increasing number of Japanese institutions have been using preoperative CRT to treat locally advanced rectal cancer. In this review, we describe the latest trends in CRT under five headings: short-course or long-course radiation, efforts to improve combined chemotherapy, the addition of preoperative adjuvant chemotherapy, the watch and wait strategy, and the significance of lateral lymph node dissection in patients receiving CRT.


Two new subspecies of Photorhabdus luminescens, isolated from Heterorhabditis bacteriophora (Nematoda: Heterorhabditidae): Photorhabdus luminescens subsp. kayaii subsp. nov. and Photorhabdus luminescens subsp. thracensis subsp. nov.
Bacterial isolates from nematodes from Turkish soil samples were initially characterized by molecular methods and seven members of the genus Photorhabdus identified to the species level, using riboprint analyses and metabolic properties. Strain 07-5 (DSM 15195) was highly related to the type strain of Photorhabdus luminescens subsp. laumondii DSM 15139T, and was regarded a strain of this subspecies. Strains 1121T (DSM 15194T), 68-3 (DSM 15198) and 47-10 (DSM 15197) formed one, strain 39-8T (DSM 15199T), 39-7 (DSM 15196) and 01-12 (DSM 15193) formed a second cluster that branched intermediate the three subspecies of Photorhabdus luminescens. Based upon moderate 16S rRNA gene sequence similarities and differences in metabolic properties among themselves and with type strains of the three subspecies we consider the two clusters to represent two new subspecies of Photorhabdus luminescens for which the names Photorhabdus luminescens subsp. kayaii, type strain 1121T (DSM 15194T, NCIMB 13951T), and Photorhabdus luminescens subsp. thracensis subsp. nov., type strain 39-8T (DSM 15199T, NCIMB 13952T) are proposed.


[The ways of purulent spread in necrotizing pancreatitis].
Results of examination of 206 patients with destructive pancreatitis are presented. On the basis of complex research it has been allocated five most significant variants of distribution of purulent process to retroperitoneal space. Existence of correlation of morphological changes in pancreas and retroperitoneal cellular tissue has been shown. Contrast ductography allowed to identify more precisely a place of defect in a wall of a duct and to predict the ways of purulent spread in retroperitoneal space. In view of it surgical tactics allows to improve results of treatment.


Transurethral light delivery for prostate photoacoustic imaging.
Photoacoustic imaging has broad clinical potential to enhance prostate cancer detection and treatment, yet it is challenged by the lack of minimally invasive, deeply penetrating light delivery methods that provide sufficient visualization of targets (e.g., tumors, contrast agents, brachytherapy seeds). We constructed a side-firing fiber prototype for transurethral photoacoustic imaging of prostates with a dual-array (linear and curvilinear) transrectal ultrasound probe. A method to calculate the surface area and, thereby, estimate the laser fluence at this fiber tip was derived, validated, applied to various design parameters, and used as an input to three-dimensional Monte Carlo simulations. Brachytherapy seeds implanted in phantom, ex vivo, and in vivo canine prostates at radial distances of 5 to 30 mm from the urethra were imaged with the fiber prototype transmitting 1064 nm wavelength light with 2 to 8 mJ pulse energy. Prebeamformed images were displayed in real time at a rate of 3 to 5 frames per second to guide fiber placement and beamformed offline. A conventional delay-and-sum beamformer provided decreasing seed contrast (23 to 9 dB) with increasing urethra-to-target distance, while the short-lag spatial coherence beamformer provided improved and relatively constant seed contrast (28 to 32 dB) regardless of distance, thus improving multitarget visualization in single and combined curvilinear images acquired with the fiber rotating and the probe fixed. The proposed light delivery and beamforming methods promise to improve key prostate cancer detection and treatment strategies.


A Systematic Review to Identify Screening Tools and Practices that Can Be Used by Children's Rehabilitation Service Providers to Screen Parents' Mental Health.
Parents of children with disabilities often report stress, depression, and anxiety. This review identified screening tools and practices that pediatric rehabilitation service providers can use to screen the mental health of parents of children with disabilities. An interdisciplinary team and patient partner completed the systematic review in which 16,015 articles were screened and 473 articles were included to i) identify mental health tools that were used with parents, ii) determine the clinical utility of frequently used tools, iii) examine the screening practices used in pediatric rehabilitation contexts. 115 screening tools were used to screen parents' mental health. The Parenting Stress Index was used most often. Seven studies reported screening in order to recommend further assessment or supports. Increased awareness, training, resources, and infrastructure are needed to support parents' mental health. Evidence is needed to guide mental health screening practices in pediatric rehabilitation and determine their effectiveness.


Delayed self-regulation and time-dependent chemical drive leads to novel states in epigenetic landscapes.
The epigenetic pathway of a cell as it differentiates from a stem cell state to a mature lineage-committed one has been historically understood in terms of Waddington's landscape, consisting of hills and valleys. The smooth top and valley-strewn bottom of the hill represent their undifferentiated and differentiated states, respectively. Although mathematical ideas rooted in nonlinear dynamics and bifurcation theory have been used to quantify this picture, the importance of time delays arising from multistep chemical reactions or cellular shape transformations have been ignored so far. We argue that this feature is crucial in understanding cell differentiation and explore the role of time delay in a model of a single-gene regulatory circuit. We show that the interplay of time-dependent drive and delay introduces a new regime where the system shows sustained oscillations between the two admissible steady states. We interpret these results in the light of recent perplexing experiments on inducing the pluripotent state in mouse somatic cells. We also comment on how such an oscillatory state can provide a framework for understanding more general feedback circuits in cell development.


Arthritis of the temporomandibular joints.
The most common disease of the temporomandibular joint (TMJ) is osteoarthritis. Rheumatoid arthritis and psoriatic arthritis may also involve this joint. Other diseases that may occasionally affect the TMJ include familial Mediterranean fever, systemic lupus erythematosus, gout, Sjögren's syndrome and infectious arthritis. Many cases of TMJ syndrome are labeled as idiopathic facial pain syndrome, a category that probably represents a number of different entities. The role of dental malocclusion has been greatly overemphasized in the past.


Production and characterization of an alkaline thermostable crude lipase from an isolated strain of Bacillus cereus C(7).
A bacterial strain isolated from spoiled coconut and identified as Bacillus cereus was found capable of producing alkaline thermostable extracellular lipase. Optimum temperature, time, and pH for enzyme substrate reaction were found to be 60 degrees C, 10 min, and 8.0 respectively. Common surfactants except Triton X 100 and cetyltrimethylammonium bromide have no or very little inhibitory effects on enzyme activity. The enzyme was found to be stable in presence of oxidizing agents and protease enzyme. The maximum lipase production was achieved at 30-33 degrees C, pH 8.0 on 24 h of fermentation using 50 ml medium in a 250-ml Erlenmeyer flask. The superior carbon and nitrogen sources for lipase production were starch (2%) and ammonium sulfate (nitrogen level 21.2 mg/100 ml), peptone (nitrogen level 297 mg/100 ml), and urea (nitrogen level 46.62 mg/100 ml) in combination, respectively. The maximum enzyme activity obtained was 33 +/- 0.567 IU/ml.


Antiproliferative Activity of Carnosic Acid is Mediated via Inhibition of Cell Migration and Invasion, and Suppression of Phosphatidylinositol 3-Kinases (PI3K)/AKT/Mammalian Target of Rapamycin (mTOR) Signaling Pathway.
BACKGROUND Lung cancer is one of the leading causes of cancer-related mortalities worldwide and majority of these deaths result from non-small cell lung cancer (NSCLC). The primary objective of this research was to determine the anticancer potential of carnosic acid, a plant derived abietane diterpene, against human lung cancer cells, as well as to determine its effects on cell migration and invasion, apoptosis, and the PI3K/AKT/m-TOR signaling pathway. MATERIAL AND METHODS Cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay; fluorescence microscopy using acridine orange/ethidium bromide stain and Comet assay were used to study cellular apoptosis. In vitro wound healing assay was used to study effects on cell migration; Transwell assay was used to study cell invasion after drug treatment. Western blot assay was used to study effects of carnosic acid on the PI3K/AKT/m-TOR signaling pathway. RESULTS It was shown that carnosic acid could inhibit the growth of A-549 human non-small cell lung carcinoma cells dose-dependently showing an IC₅₀ value of 12.5 μM. This growth inhibition of A-549 cells was mediated via apoptotic cell death as observed by fluorescence microscopy showing nuclear fragmentation and chromatin condensation. Carnosic acid, dose-dependently, also inhibited cell migration and invasion. Finally, western blot assay revealed that carnosic acid also led to inhibition of the PI3K/AKT/m-TOR signaling pathway. CONCLUSIONS In conclusion, our results showed that Carnosic acid has the potential to inhibit cancer cell growth in A-549 lung cancer cells by activating apoptotic death, inhibiting cell migration and invasion and suppressing PI3K/AKT/m-TOR signaling pathway.


Octopine as an end product of anaerobic glycolysis in the chambered nautilus.
The terminal step in the anaerobic glycolysis of muscle in the chambered nautilus, Nautilus pompilius, is not pyruvate reduction to lactate as in vertebrate muscle. Instead of lactate dehydrogenase, these organisms utilize octopine dehydrogenase (E.C. 188.8.131.52), catalyzing the reductive condensation of pyruvate and arginine, which is dependent on the reduced form of nicotinamide adenine dinucleotide, to form octopine and the oxidized form of the coenzyme. The kinetic properties of octopine dehydrogenase favor the production of octopine, which accumulates under a variety of conditions.


Neuropeptides degranulate serous cells of ferret tracheal glands.
To determine whether serous or mucous cells in tracheal submucosal glands respond to the neuropeptides substance P (SP) and vasoactive intestinal peptide (VIP), we studied the peptide-induced changes in gland cell morphology accompanying release of 35SO4-labeled macromolecules from tracheal explants of ferrets. Explants were labeled for 1 h in medium containing 35SO4 and washed for 3.5 additional hours. Base-line secretion in the absence of drugs declined between 1.5 and 3.5 h after the pulse. Between 2.5 and 3.5 h, the average percent change in counts per minute recovered per sample period was not significantly different from zero (P greater than 0.3; n = 6). Substance P (10(-5) M) and VIP (2 X 10(-6) M) added 4 h after labeling each increased greatly the release of 35SO4-labeled macromolecules (SP, 219%; VIP, 180%) above base line. Bethanechol, a muscarinic-cholinergic agonist (10(-5) M), increased secretion by an average of 142% above base line (each effect, P less than 0.05; n = 6 each). Light and electron microscopy of the control tissues showed glands with narrow lumens and numerous secretory granules. Glands treated with SP or VIP had enlarged lumens and the serous cells were markedly degranulated. These phenomena were documented by morphometry and suggest that SP and VIP cause secretion from glands at least partially by stimulating exocytosis from serous cells.


Continuation-based numerical detection of after-depolarization and spike-adding thresholds.
The changes in neuronal firing pattern are signatures of brain function, and it is of interest to understand how such changes evolve as a function of neuronal biophysical properties. We address this important problem by the analysis and numerical investigation of a class of mechanistic mathematical models. We focus on a hippocampal pyramidal neuron model and study the occurrence of bursting related to the after-depolarization (ADP) that follows a brief current injection. This type of burst is a transient phenomenon that is not amenable to the classical bifurcation analysis done, for example, for periodic bursting oscillators. In this letter, we show how to formulate such transient behavior as a two-point boundary value problem (2PBVP), which can be solved using well-known continuation methods. The 2PBVP is formulated such that the transient response is represented by a finite orbit segment for which onsets of ADP and additional spikes in a burst can be detected as bifurcations during a one-parameter continuation. This in turn provides us with a direct method to approximate the boundaries of regions in a two-parameter plane where certain model behavior of interest occurs. More precisely, we use two-parameter continuation of the detected onset points to identify the boundaries between regions with and without ADP and bursts with different numbers of spikes. Our 2PBVP formulation is a novel approach to parameter sensitivity analysis that can be applied to a wide range of problems.


Melanin pigment of Streptomyces puniceus RHPR9 exhibits antibacterial, antioxidant and anticancer activities.
The present study reveals the production of dark, extracellular melanin pigment (386 mg/L) on peptone yeast extract iron agar medium by Streptomyces puniceus RHPR9 using the gravimetric method. UV-Visible, Fourier Transform Infrared (FTIR), and Nuclear Magnetic Resonance (1H) (NMR) spectroscopy confirmed the presence of melanin. Extracted melanin showed antibacterial activity against human pathogens such as Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli except for Klebsiella pneumoniae. A potent free radical scavenging activity was observed at 100 μg/mL of melanin by the DPPH method with a concentration of 89.01±0.05% compared with ascorbic acid 96.16±0.01%. Antitumor activity of melanin was evaluated by MTT assay against HEK 293, HeLa, and SK-MEL-28 cell lines with IC50 values of 64.11±0.00, 14.43±0.02, and 13.31±0.01 μg/mL respectively. Melanin showed maximum anti-inflammatory activity with human red blood cells (hRBC) (78.63 ± 0.01%) and minimum hemolysis of 21.37±0.2%. The wound healing potential of the pigment was confirmed on HeLa cells, cell migration was calculated, and it was observed that cell migration efficiency decreased with an increase in the concentration of melanin. To our knowledge, this is the first evidence of melanin produced from S. puniceus RHPR9 that exhibited profound scavenging, anti-inflammatory and cytotoxic activities.


Failure of BRCA1 dysfunction to alter ovarian cancer survival.
Many factors modify ovarian cancer survival. There are conflicting reports regarding survival of individuals with hereditary BRCA1-related ovarian cancer. None have controlled for other mechanisms of BRCA1 silencing in the control cohort. Fifty-nine cancers with presumed BRCA1 dysfunction because of mutation (24 germ-line and 16 somatic) or absent BRCA1 mRNA because of promoter hypermethylation (n = 19) were identified among 250 consecutively screened ovarian cancers. Controls were matched from the same population based on p53 mutation type, age at diagnosis, Fédération Internationale des Gynaecologistes et Obstetristes surgical stage and histological grade, residual disease, preoperative CA125, disease site, and the presence of BRCA1 mRNA translatable in an in vitro protein expression assay. BRCA1 promoter hypermethylation was determined by the methylation-specific PCR technique. The significance of promoter hypermethylation was confirmed by the absence of detectable BRCA1 mRNA. The median survival for individuals with ovarian cancer BRCA1 dysfunction was 4.1 years versus 3.5 years in the case matched controls (P = 0.98). Grouped on the basis of the mechanism of BRCA1 dysfunction, median survival was 4.5, 2.8, and 2.3 years for germ-line, somatic, and BRCA1 promoter-silenced ovarian cancers. However, for the corresponding matched controls with wild-type BRCA1 sequence, the median survival was virtually identical: 4.6, 2.8, and 3.3 years, respectively. In a Cox proportional hazards analysis, only residual disease (P = 0.0001), age (P = 0.01), and Fédération Internationale des Gynaecologistes et Obstetristes stage (P = 0.011) entered the survival model. In contrast with other published reports, we are unable to detect large survival differences between matched case-control cohorts of ovarian cancers with BRCA1 inactivation by any of three independent mechanisms.


Thyroid Deficiency Before Birth Alters the Adipose Transcriptome to Promote Overgrowth of White Adipose Tissue and Impair Thermogenic Capacity.
Background: Development of adipose tissue before birth is essential for energy storage and thermoregulation in the neonate and for cardiometabolic health in later life. Thyroid hormones are important regulators of growth and maturation in fetal tissues. Offspring hypothyroid in utero are poorly adapted to regulate body temperature at birth and are at risk of becoming obese and insulin resistant in childhood. The mechanisms by which thyroid hormones regulate the growth and development of adipose tissue in the fetus, however, are unclear. Methods: This study examined the structure, transcriptome, and protein expression of perirenal adipose tissue (PAT) in a fetal sheep model of thyroid hormone deficiency during late gestation. Proportions of unilocular (UL) (white) and multilocular (ML) (brown) adipocytes, and UL adipocyte size, were assessed by histological and stereological techniques. Changes to the adipose transcriptome were investigated by RNA sequencing and bioinformatic analysis, and proteins of interest were quantified by Western blotting. Results: Hypothyroidism in utero resulted in elevated plasma insulin and leptin concentrations and overgrowth of PAT in the fetus, specifically due to hyperplasia and hypertrophy of UL adipocytes with no change in ML adipocyte mass. RNA sequencing and genomic analyses showed that thyroid deficiency affected 34% of the genes identified in fetal adipose tissue. Enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathways were associated with adipogenic, metabolic, and thermoregulatory processes, insulin resistance, and a range of endocrine and adipocytokine signaling pathways. Adipose protein levels of signaling molecules, including phosphorylated S6-kinase (pS6K), glucose transporter isoform 4 (GLUT4), and peroxisome proliferator-activated receptor γ (PPARγ), were increased by fetal hypothyroidism. Fetal thyroid deficiency decreased uncoupling protein 1 (UCP1) protein and mRNA content, and UCP1 thermogenic capacity without any change in ML adipocyte mass. Conclusions: Growth and development of adipose tissue before birth is sensitive to thyroid hormone status in utero. Changes to the adipose transcriptome and phenotype observed in the hypothyroid fetus may have consequences for neonatal survival and the risk of obesity and metabolic dysfunction in later life.


Love at first fright: partner salience moderates roller-coaster-induced excitation transfer.
This study examined the effects of residual nervous system arousal on perceptions of sexual attraction. Researchers approached individuals (males, n = 165; females, n = 135) at amusement parks as they were either waiting to begin or as they had just gotten off a roller-coaster ride. Participants were shown a photograph of an average attractive, opposite-gendered individual and asked to rate the individual on attractiveness and dating desirability. Participants were also asked to rate their seatmates' levels of attractiveness. Consistent with the predictions of excitation transfer theory, for males and females riding with a nonromantic partner, ratings of attractiveness and dating desirability toward the photographed individual were higher among persons exiting than entering the ride. Among persons riding with a romantic partner, there were no significant differences in attractiveness or dating desirability ratings between persons entering and exiting the ride. The findings are discussed in terms of the potential moderator effects of a salient romantic partner on excitation transfer.


Management of pregnancy in women with factor VII deficiency: A case series.
Inherited factor VII deficiency is the most common autosomal recessive inherited bleeding disorder, with an estimated incidence of one per 500 000 cases in the general population. Bleeding manifestations correlate poorly with circulating FVII levels. During pregnancy, increases in FVII levels can occur in women with mild-moderate FVII deficiencies but not in those with severe deficiency. We present five pregnant patients with FVII deficiency and describe the management during their pregnancies and peripartum periods. Retrospective analysis of six pregnancies in five women with FVII deficiency followed during pregnancy and delivery at an academic medical centre between January 2013 and December 2019. Of the five patients, two had severe, one with moderate and two with mild FVII deficiency. Early postpartum haemorrhage (PPH) occurred in two patients. One of the two severe FVII-deficient patients had PPH with a laceration at delivery despite replacement therapy with recombinant factor VII. The other PPH occurred in a patient with mild FVII deficiency who delivered twins by caesarean section under general anaesthesia. Neuraxial anaesthesia was utilized in only one woman with mild deficiency whose FVII level normalized at the end of the pregnancy. Management of delivery for women with FVII deficiency should be addressed on a case-by-case basis at centres with expertise in rare bleeding disorders, maternal foetal medicine and obstetric anaesthesiology. These management discussions should factor the patient's bleeding history, third trimester PT, FVII level, multiple gestation and mode of delivery.


Deletions at 22q11.2 in idiopathic Parkinson's disease: a combined analysis of genome-wide association data.
Parkinson's disease has been reported in a small number of patients with chromosome 22q11.2 deletion syndrome. In this study, we screened a series of large, independent Parkinson's disease case-control studies for deletions at 22q11.2. We used data on deletions spanning the 22q11.2 locus from four independent case-control Parkinson's disease studies (UK Wellcome Trust Case Control Consortium 2, Dutch Parkinson's Disease Genetics Consortium, US National Institute on Aging, and International Parkinson's Disease Genomics Consortium studies), which were independent of the original reports of chromosome 22q11.2 deletion syndrome. We did case-control association analysis to compare the proportion of 22q11.2 deletions found, using the Fisher's exact test for the independent case-control studies and the Mantel-Haenszel test for the meta-analyses. We retrieved clinical details of patients with Parkinson's disease who had 22q11.2 deletions from the medical records of these patients. We included array-based copy number variation data from 9387 patients with Parkinson's disease and 13 863 controls. Eight patients with Parkinson's disease and none of the controls had 22q11.2 deletions (p=0·00082). In the 8451 patients for whom age at onset data were available, deletions at 22q11.2 were associated with Parkinson's disease age at onset (Mann-Whitney U test p=0·001). Age at onset of Parkinson's disease was lower in patients carrying a 22q11.2 deletion (median 37 years, 95% CI 32·0-55·5; mean 42·1 years [SD 11·9]) than in those who did not carry a deletion (median 61 years, 95% CI 60·5-61·0; mean 60·3 years [SD 12·8]). A 22q11.2 deletion was present in more patients with early-onset (p<0·0001) and late-onset Parkinson's disease (p=0·016) than in controls, and in more patients with early-onset than late-onset Parkinson's disease (p=0·005). Clinicians should be alert to the possibility of 22q11.2 deletions in patients with Parkinson's disease who have early presentation or features associated with the chromosome 22q11.2 deletion syndrome, or both. UK Medical Research Council, UK Wellcome Trust, Parkinson's UK, Patrick Berthoud Trust, National Institutes of Health, "Investissements d'Avenir" ANR-10-IAIHU-06, Dutch Parkinson Foundation (Parkinson Vereniging), Neuroscience Campus Amsterdam, National Institute for Health Research, National Institute on Aging, National Institutes of Health.


Precision radiotherapy using MR-linac for pancreatic neuroendocrine tumors in MEN1 patients (PRIME): a protocol for a phase I-II trial, and systematic review on available evidence for radiotherapy of pNETs.
Surgical resection is the standard of care for the treatment of pancreatic neuro-endocrine tumors (pNETs) in patients with Multiple Endocrine Neoplasia Type 1 (MEN1). However, surgery can cause significant short- and long-term morbidity. Magnetic resonance-guided radiotherapy (MRgRT) is a potential effective treatment with little side effects. With traditional radiotherapy techniques, irradiation of pancreatic tumors to high dose levels was hampered by poor visibility of the tumor during treatment. MRgRT uses onboard MRI to guide the treatment, thereby enabling delivery of ablative irradiation doses to the tumor, while sparing surrounding tissues. In this study, we describe results from a systematic review assessing efficacy of radiotherapy in pNET and present the protocol of the PRIME study. PubMed, Embase and Cochrane Library were searched for articles assessing efficacy and side effects of radiotherapy for the treatment of pNETs. Risk of bias was assessed using the ROBINS-I Risk of Bias Tool for observational studies. Descriptive statistics were used to describe results of included trials. Four studies comprising of 33 patients treated by conventional radiotherapy were included. Despite the heterogeneity of studies, radiotherapy appeared to be effective for the treatment of pNETs with most patients responding (45.5%) or stabilizing (42.4%) in tumor size. Due to the limited literature available and concerns about damage to surrounding tissue, conventional radiotherapy is currently little used for pNETs. The PRIME study is a phase I-II trial with a single arm prospective cohort study design, investigating the efficacy of MRgRT in MEN1 patients with pNET. MEN1 patients with growing pNETs with a size between 1.0 and 3.0 cm without malignant features are eligible for inclusion. Patients are treated with 40 Gy in 5 fractions on the pNET, using online adaptive MRgRT on a 1.5T MR-linac. The primary endpoint is the change in tumor size at MRI 12 months follow-up. Secondary endpoints include radiotoxicity, quality of life, endocrine and exocrine pancreas function, resection rate, metastatic free and overall survival. When MRgRT is found effective with low radiotoxicity, it could reduce the need for surgery for pNET and preserve quality of life. PROSPERO https://clinicaltrials.gov/, (CRD42022325542).


Pathology and Case Definition of Severe Perkinsea Infection of Frogs.
Severe Perkinsea infection (SPI) is an emerging disease of frogs responsible for mass mortalities of tadpoles across the United States. It is caused by protozoa belonging to the phylum Perkinsozoa that form a distinct group referred to as the Pathogenic Perkinsea Clade of frogs. In this work, we provide detailed description of gross and histologic lesions from 178 naturally infected tadpoles, including 10 species from 22 mortality events and 6 amphibian health monitoring studies from diverse geographic areas. On external examination, we observed abdominal distension (10, 5.6%), cutaneous erythema and petechia (3, 1.7%), subcutaneous edema (3, 1.7%), and areas of white skin discoloration (3, 1.7%). On macroscopic examination of internal organs, we found hepatomegaly (68, 38.2%), splenomegaly (51, 28.7%), nephromegaly (47, 26.4%), ascites (15, 8.4%), segmental irregular thickening and white discoloration of the intestine (8, 4.5%), pancreatomegaly (4, 2.2%), and pancreatic petechia (1, 0.6%). Histologically, over 60% of the liver (148/165, 89.7%), kidney (113/147, 76.9%), spleen (96/97, 99%), and pancreas (46/68, 67.6%) were invaded by myriad intracellular and extracellular Perkinsea hypnospore-like and trophozoite-like organisms. Numerous other tissues were affected to a lesser extent. Mild histiocytic inflammation with fewer lymphocytes or eosinophils was commonly observed in areas of infection that were not obscured by lympho-granulocytic hematopoietic tissue. In light of these observations, we suggest a logical pathogenesis sequence. Finally, we propose a "case definition" for SPI to promote standardized communication of results and prevent misdiagnosis with epidemiological and pathologically overlapping diseases such as ranavirosis.


The Ocular Microbiome: Molecular Characterisation of a Unique and Low Microbial Environment.
Aim: The ocular surface is continually exposed to bacteria from the environment and traditional culture-based microbiological studies have isolated a low diversity of microorganisms from this region. The use of culture-independent methods to define the ocular microbiome, primarily involving 16S ribosomal RNA gene sequencing studies, have shown that the microbial communities present on the ocular surface have a greater diversity than previously reported. Method: A review of the literature on ocular microbiome research in health and disease. Results: Molecular techniques have been used to investigate the effect of contact lens wear and disease on the microbiota of the ocular surface and eyelids and the immunoregulatory role of the ocular surface microbiota. Studies have shown that compositional changes in the microbiota occur in ocular surface disorders such as blepharitis, trachoma and dry eye and also suggest a role of the ocular and non-ocular microbiome in retinal disease including age-related macular degeneration, glaucoma, uveitis and diabetic retinopathy. However, ocular microbiome studies need to recognise the potential for contamination to impact findings and carefully control each stage of the experimental procedure and to utilise statistical methods to identify contamination signals. Conclusion: The healthy ocular surface is characterised by a relatively stable, comparatively low diversity microbiome with recent findings that the bacteria of the ocular surface appear to have a role in maintaining homeostasis by modulating immune function.


An improved method of nasojejunal feeding tube placement for patients requiring endoscopic nasobiliary drainage.
To avoid a second endoscopy for nasojejunal feeding tube placement (NFTP) in patients undergoing endoscopic nasobiliary drainage (ENBD), we studied improved NFTP method and compared it to endoscopic method. Patients with ENBD were divided into two groups. One group (18 patients) received endoscopic NFTP and the other group (26 patients) received improved NFTP. Placement time, physical condition of the patients and complications were recorded. In 18 patients who underwent endoscopic NFTP, NFT was successfully placed on the first attempt in 14 patients with a first placement success rate of 77.8%. NFT was wrongly intubated into the trachea in one patient inducing coughing, and after it was removed, the second placement was successful. The total success rate of endoscopic NFTP was 83.3% with an average placement time of 17.0 minutes. In 26 patients undergoing improved NFTP, all were successfully placed on the first attempt with a success rate of 100%, and an average placement time of 2.55 minutes. In patients with ENBD, the success rate of improved NFTP was significantly higher than endoscopic NFTP (χ²=36.4, p<0.05) with a significantly shorter placement time (t=18.5, p<0.05). For patients with ENBD, improved NFTP method is superior to the endoscopic method as it is more effective, convenient, faster, and cheaper. Additionally it avoids a second endoscopy and has fewer complications, better security and a higher success rate. The improved method is a safer, easier, more effective and practical method of EN and deserves general adoption in clinical work.


[Refractory eosinophilic gastroenteritis with gastritis, duodenal ulcer, and colitis: a case report].
Eosinophilic gastroenteritis is a rare disease that causes various abdominal symptoms that result from the infiltration of eosinophils in the digestive tract. However, this condition has been poorly explored, and the treatment criteria and prognosis after treatment are still unclear. A 20-year-old man with a refractory duodenal ulcer had been undergoing treatment since 14 years of age at another hospital. He was admitted to our hospital with abdominal pain and anemia (hemoglobin:6.3g/dL). His blood test showed elevated serum immunoglobulin E levels, considering that he was allergic to many foods. Furthermore, endoscopic biopsy detected the occurrence of eosinophilic gastroenteritis with gastritis, duodenal ulcer, and colitis. We treated him by avoiding allergenic foods and prescribing antihistamine and vonoprazan;however, duodenal ulcer and gastrointestinal tract inflammation did not show improvement. Thus, he was diagnosed with wide-ranging and refractory eosinophilic gastroenteritis and treated with 40mg/day of steroids. After 2 months, he recovered from gastritis and duodenal ulcer, and his eosinophil level decreased, as assessed using endoscopic biopsy. Eosinophilic gastroenteritis is poorly investigated, and its treatment standards have not yet been determined. Nonetheless, steroid treatment is often applied in severe cases.


Lianas suppress seedling growth and survival of 14 tree species in a Panamanian tropical forest.
Lianas are a common plant growth form in tropical forests, where they compete intensely with trees, decreasing tree recruitment, growth, and survival. If the detrimental effects of lianas vary significantly with tree species identity, as is often assumed, then lianas may influence tree species diversity and community composition. Furthermore, recent studies have shown that liana abundance and biomass are increasing relative to trees in neotropical forests, which will likely magnify the detrimental effects of lianas and may ultimately alter tree species diversity, relative abundances, and community composition. Few studies, however, have tested the responses of multiple tree species to the presence of lianas in robust, well-replicated experiments. We tested the hypotheses that lianas reduce tree seedling growth and survival, and that the effect of lianas varies with tree species identity. We used a large-scale liana removal experiment in Central Panama in which we planted 14 replicate seedlings of 14 different tree species that varied in shade tolerance in each of 16 80 x 80 m plots (eight liana-removal and eight unmanipulated controls; 3136 total seedlings). Over a nearly two-yr period, we found that tree seedlings survived 75% more, grew 300% taller, and had twice the aboveground biomass in liana-removal plots than seedlings in control plots, consistent with strong competition between lianas and tree seedlings. There were no significant differences in the response of tree species to liana competition (i.e., there was no species by treatment interaction), indicating that lianas had a similar negative effect on all 14 tree species. Furthermore, the effect of lianas did not vary with tree species shade tolerance classification, suggesting that the liana effect was not solely based on light. Based on these findings, recently observed increases in liana abundance in neotropical forests will substantially reduce tree regeneration, but will not significantly alter tropical tree species diversity, relative abundance, or community composition.


Key factors of clinical research network capacity building.
In general, clinical research network capacity building refers to programs aimed at enhancing networks of researchers to conduct clinical research. Although in the literature there is a large body of research on how to develop and build capacity in clinical research networks, the conceptualizations and implementations remain controversial and challenging. Moreover, the experiences learnt from the past accomplishments and failures can assist in the future capacity building efforts to be more practical, effective and efficient. In this paper, we aim to provide an overview of capacity building in clinical research network by (1) identifying the key barriers to clinical research network capacity building, (2) providing insights into how to overcome those obstacles, and (3) sharing our experiences in collaborating with national and international partners to build capacity in clinical research networks. In conclusion, we have provided some insight into how to address the key factors of clinical research network capacity building and shared some empirical experiences. A successful capacity building practice requires a joint endeavor to procure sufficient resources and support from the relevant stakeholders, to ensure its efficiency, cost-effectiveness, and sustainability.


Critical comparison of image analysis workflows for quantitative cell morphological evaluation in assessing cell response to biomaterials.
Quantitative image analysis is an important tool in understanding cell fate processes through the study of cell morphological changes in terms of size, shape, number, and orientation. In this context, this work explores systematically the main challenges involved in the quantitative analysis of fluorescence microscopy images and also proposes a new protocol while comparing its outcome with the widely used ImageJ analysis. It is important to mention that fluorescence microscopy is by far most widely used in biocompatibility analysis (observing cell fate changes) of implantable biomaterials. In this study, we employed two different image analyses toolsets: (a) the conventionally employed ImageJ software, and (b) a recently developed automated digital image analyses framework, called ImageMKS. While ImageJ offers a powerful toolset for image analyses, it requires sophisticated user expertise to design and iteratively refine the analyses workflow. This workflow primarily comprises a sequence of image transformations that typically involve de-noising and labeling of features. On the other hand, ImageMKS automates the image analyses protocol to a large extent, and thereby mitigates the influence of the user bias on the final results. This aspect is addressed using a case study of C2C12 mouse myoblast cells grown on poly(vinylidene difluoride) (PVDF) based polymeric substrates. In particular, we used a number of fluorescence microscopy images of these mouse myoblasts grown on PVDF-based nanobiocomposites under the influence of electric field. In addition to the MKS workflows requiring much less user time because of their automation, it was observed that ImageMKS workflows consistently produced more reliable results that correlated better with the previously reported experimental studies.


High Prevalence of Celiac Disease Among Screened First-Degree Relatives.
To investigate the prevalence of first-degree relatives (FDRs) with celiac disease detected at screening and diagnostic significance of anti-tissue transglutaminase (anti-TTG). We performed a retrospective cohort study of 104 patients with a diagnosis of celiac disease and their FDRs, collecting data from electronic records of Mayo Clinic and celiac disease registry from December 20, 1983, to May 22, 2017. We collected demographics, presenting symptoms, indication for testing, family history, number of other family members screened, biopsy reports, and results of serologic tests. Of 477 FDRs identified, 360 were screened (mean screening rate per family, 79%±25%) and 160 FDRs (44.4%) were diagnosed with celiac disease, at a mean age 31.9±21.6 years (62% female). All diagnosed FDRs had positive anti-TTG titers. Clinical features were documented in 148 diagnosed FDRs, of those 9 (6%) had classic, 97 (66%) had non-classic symptoms, and 42(28%) had no reported symptoms. Histology reports were available from 155 FDRs: 12 (8%) had Marsh 1, 77 (50%) had Marsh 3a, and 66 (43%) had Marsh 3b. A level of anti-TTG greater than or equal to 2.75 of the upper limit of normal identified FDRs with villous atrophy with 87% sensitivity, 82% specificity, and a positive predictive value of 95%. In a retrospective cohort study of patients diagnosed with celiac disease, we found a high prevalence of celiac disease among screened FDRs. High anti-TTG titers associated with villous atrophy on small bowel biopsies, irrespective of symptoms.


Anxiety in 3- to 7-year-old children with autism spectrum disorder seeking treatment for disruptive behavior.
Anxiety is a common and impairing problem in children with autism spectrum disorder, but little is known about it in preschool children with autism spectrum disorder. This article reports on the characteristics of anxiety symptoms in young children with autism spectrum disorder using a parent-completed rating scale. One hundred and eighty children (age 3-7 years) participated in a clinical trial of parent training for disruptive behaviors. Anxiety was measured as part of pre-treatment subject characterization with 16 items from the Early Childhood Inventory, a parent-completed scale on child psychiatric symptoms. Parents also completed other measures of behavioral problems. Sixty-seven percent of children were rated by their parents as having two or more clinically significant symptoms of anxiety. There were no differences in the Early Childhood Inventory anxiety severity scores of children with IQ < 70 and those with ⩾70. Higher levels of anxiety were associated with severity of oppositional defiant behavior and social disability. Anxiety symptoms are common in preschoolers with autism spectrum disorder. These findings are consistent with earlier work in school-age children with autism spectrum disorder. There were no differences in anxiety between children with IQ below 70 and those with IQ of 70 and above. Social withdrawal and oppositional behavior were associated with anxiety in young children with autism spectrum disorder.


Hearing Outcomes After Surgical Manipulation of the Membranous Labyrinth During Superior Semicircular Canal Dehiscence Plugging or Posterior Semicircular Canal Occlusion.
To analyze audiometric data after surgical manipulation of the membranous labyrinth during plugging of superior semicircular canal dehiscence (SSCD) or posterior semicircular canal occlusion (PSCO) for benign paroxysmal positional vertigo. Retrospective chart review. Tertiary referral center. Patients undergoing plugging of SSCD or PSCO between 2009 and 2019. Pre- and postoperative audiometric data were collected per AAO-HNS guidelines. Hearing outcomes at initial and last follow-up were compared. Subanalyses were performed for surgical approach and age. Eighty-seven total procedures in 76 patients including 43 middle cranial fossa for SSCD, 29 transmastoid SSCD, and 15 PSCO. Mean preoperative air-conduction-pure-tone averages was 21.1±14.9 dB compared with 26.1 ± 19.6 dB at initial follow-up and 24.4 ± 18.6 dB at last follow-up (p = 0.006). Mean preoperative bone-conduction-pure-tone average was 14.3 ± 11.9 dB compared with 18.3 ± 15.6 dB at initial follow-up and 18.5 ± 16.9 dB at last follow-up (p < 0.001). There were five cases of hearing loss >20 dB including one case of profound sensorineural hearing loss >55 dB. PSCO resulted in the most hearing loss at initial follow-up but largely resolves with time. Transmastoid approaches for SSCD resulted in more hearing loss compared with middle cranial fossa. Hearing outcomes were generally stable for SSCD approaches but showed improvement over time for PSCO. Age >50 was associated with greater hearing loss of 5.2 ± 11.1 dB compared with 1.3 ± 10.5 dB but did not reach statistical significance (p = 0.110). Surgical manipulation of the membranous labyrinth results in statistically significant hearing loss in a pooled analysis. Transient hearing loss is observed in PSCO and TM SSCD plugging was associated with postoperative hearing loss. There was a trend toward increased hearing loss in patients >50 years old.


First Report of Segmented Filamentous Bacteria Associated with Rhigonema Sp. (Nematoda: Rhigonematidae) Dwelling in Hindgut of Riukiaria Sp. (Diplopoda: Xystodesmidae).
We morphologically and molecularly characterized segmented filamentous bacteria (SFB) associated with Rhigonema sp. nematodes in millipede hindguts. Seventy-three Riukiaria sp. millipedes were collected from a broad-leaf forest in Japan, and nematodes were excised from the millipede's hindguts. The occurrence rate of SFB associated with nematodes was 24 % (10/41) for males, 47 % (14/30) for females, and 100 % (2/2) for juveniles. Genomic DNA was extracted from four SFB-rich nematode heads, and we obtained 40 bacterial clones via analysis of nearly full-length 16S rDNA gene sequences. At the phylum level, Firmicutes, Proteobacteria, and Verrucomicrobia accounted for 55 %, 40 %, and 5 % of SFB, respectively. In Firmicutes, Clostridiaceae (28 %) and Lachnospiraceae (15 %) were the dominant groups. Our sequences were divided into seven and three subclades between Firmicutes and Proteobacteria in the phylogenetic tree. In the Firmicutes clade, eight sequences were classified as Lachnospiraceae with a bootstrap value >83 %. A phylogenetic tree involving known uncultured Lachnospiraceae sequences characterized the phylogenetic position of SFB associated with nematodes. Our results suggest that the association of SFB with nematode bodies was probably incidental and that SFB are not always present in millipede hindguts. Our bacterial groups corresponded to those of arthropod hindgut, and SFB associated with nematodes were inferred to belong to Lachnospiraceae. Because the Lachnospiraceae sequences obtained in this study showed specific lineages that differed from all the known deposited sequence data, these groups may be unique to Riukiaria sp.


Synergistic effect of curcumin and insulin on muscle cell glucose metabolism.
Curcumin is a major phenolic compound of Curcuma longa, which has long been used in traditional Indian medicine. Recently, curcumin has been reported to have antihyperglycemic activity in animal models. However, the molecular basis of this action has not been adequately described. In the present study, curcumin was observed to contain a high level of polyphenols and strong antioxidant activity. Then, the antihyperglycemic effect of curcumin was examined for various signaling pathways using C(2)C(12) mouse myoblast cells. From this, curcumin treatment strongly induced glucose uptake and the phosphorylation of AMPK (AMP-activated protein kinase)/ACC (acetyl-CoA carboxylase), but not PI3-kinase (phosphoinositide 3-kinase)/Akt. Interestingly, the co-treatment of insulin and curcumin produced a mutual synergistic activation of both AMPK/ACC and PI3-kinase/Akt pathways. On the other hand, the synergism could not be observed from the co-treatment of insulin and EGCG. It suggests that the two signaling pathways can crosstalk each other in a case-sensitive manner upon the treatment of antioxidant polyphenols. Further, these results were consistent with the findings of GLUT4 translocation to the cell surface. Our findings indicate that curcumin can promote AMPK activation and glucose uptake with increased insulin sensitivity in muscle cells as a potential anti-diabetic therapeutic agent.


[The diagnostic of defects of inborn immunity under B-cell tumors of lymphatic system].
The genetic characteristics are key risk factors of development of many human neoplasms including B-cell tumors of lymphatic system. The relationship between polymorphic variants of genes FCGR2A (His 1 66Agr), CD14 (C-159T). IL1β (T-31C), IL2 (7:330G) and 7LR2 (Arg753Ghn) and development of various forms of B-cell tumors of lymphatic system in 80 patients was investigated. The statistically significant differences of rates of particiular genotypes of single nucleotid polymorphisms of genes FCGR2A, CD14. IL1β, IL2 and TLR2 in patients with indolent and aggressive types of course of non-Hodgkin lymphoma and also multiple myeloma. The results prove hypothesis that genetic variants of genes of inborn immune response effect the origin and character of course of different types of lymphoproliferative diseases. The markers can become additional prognostic characteristics of benign and aggressive course of tumors.


Recurrent epithelial ovarian cancer: an update on treatment.
An estimated 85% of patients with epithelial ovarian cancer who achieve a full remission following first-line therapy will develop recurrent disease. Although each subsequent line of therapy is characterized by shorter disease-free intervals, median survival for these patients ranges from 12 months to 24 months. Emerging therapies in the management of ovarian cancer have resulted in a shift in paradigm, including in the appropriate time to institute therapy, and in the selection of therapy. This review focuses on chemotherapy and emerging biologic agents that present a therapeutic option for patients with recurrent ovarian cancer.


Information technology creeps into rural general practice.
This study looked at the effect of information technology on rural medical practice. Eight GPs in rural medical practice in Victoria were interviewed, and World Wide Web sites were accessed for information relevant to the rural GPs use of information technology. The results indicated that rural GPs are developing their use of information technology according to their needs. The use of information technology is changing the nature of rural medical practice, and bringing more support and information to rural GPs. However, some of the technologies now available are of little perceived use to the GPs, and GPs with good support staff are better able to take advantage of the technology than GPs with little or inadequate support. The findings of this study have implications for policies being implemented to increase the availability of information technology to support rural health care.


[Drug use in children resident in a small urban area].
The use of potentially addictive substances may be influenced by the young person's place of residence. To determine the frequency and characteristics of tobacco, alcohol and cannabis use in secondary school pupils in a small urban area during the academic year 2008-09. Cross-sectional survey. Socio-demographic data related to tobacco, alcohol and cannabis use were gathered by means of a confidential self-administered questionnaire. The survey was responded to by 88% of the pupils (393). Prevalence of tobacco use was 48%, 16% of whom smoked every day. Forty four percent of third grade pupils and fifty two per cent of forth-graders, did consume alcohol on the week ends. Thirty-five percent said that they had been drunk on at least one occasion, while 30% of the pupils had tried cannabis, and 50% of third-graders. Among those who had used cannabis, 23% had done so in the previous month. At onset of cannabis use was 13 years. Almost 80% of those surveyed said they had not paid for the first drugs they used. The use of legal and illegal drugs for those in our sample begins between the ages of 12 and 14 years, representing slightly higher frequencies than those found in big cities. Despite the illegality of the sale of tobacco and alcohol to minors, Spanish teenagers continue to obtain them. It is essential for primary care professionals, educators and families to be aware of the importance of early detection of the use of psychoactive substances. The prevention of drug addiction is more effective the sooner the potential risk is detected.


Venous Thromboembolism in Cancer Patients Undergoing Chemotherapy: Risk Factors and Prevention.
Ambulatory cancer patients undergoing chemotherapy are at a substantial risk for venous thromboembolism (VTE) over the course of therapy and beyond it. Factors accounting for this risk include the activation of extrinsic and intrinsic coagulation pathways, platelet activation, impaired fibrinolysis, use of catheters, infusion of blood products, and thrombogenic potential of several chemotherapeutic drugs. A few stratification models can help identify patients at a higher risk of chemotherapy-associated VTE, who may benefit from preventive strategies. Although low-molecular-weight heparins (LMWHs) effectively reduce the risk of VTE, current guidelines recommend against their routine use. Based on the results of recent randomized controlled clinical trials, the administration of prophylactic doses of the novel direct oral anticoagulants (DOACs) to ambulatory cancer patients undergoing chemotherapy has the potential to offer an effective and safe protection against VTE, obviating the inconveniences of heparins. Except for patients in whom the novel drugs are unsuitable or are contraindicated, in all other patients LMWHs should be replaced by low-dose DOACs.


Two distinct mechanisms shape the reliability of neural responses.
Despite intrinsic noise sources, neurons can generate action potentials with remarkable reliability. This reliability is influenced by the characteristics of sensory or synaptic inputs, such as stimulus frequency. Here we use conductance-based models to study the frequency dependence of reliability in terms of the underlying single-cell properties. We are led to distinguish a mean-driven firing regime, where the stimulus mean is sufficient to elicit continuous firing, and a fluctuation-driven firing regime, where spikes are generated by transient stimulus fluctuations. In the mean-driven regime, the stimulus frequency that induces maximum reliability coincides with the firing rate of the cell, whereas in the fluctuation-driven regime, it is determined by the resonance properties of the subthreshold membrane potential. When the stimulus frequency does not match the optimal frequency, the two firing regimes exhibit different "symptoms" of decreased reliability: reduced spike-time precision and reduced spike probability, respectively. As a signature of stochastic resonance, reliable spike generation in the fluctuation-driven regime can benefit from intermediate amounts of noise that boost spike probability without significantly impairing spike-time precision. Our analysis supports the view that neurons are endowed with selection mechanisms that allow only certain stimulus frequencies to induce reliable spiking. By modulating the intrinsic cell properties, the nervous system can thus tune individual neurons to pick out specific input frequency bands with enhanced spike precision or spike probability.


The expression of acylphosphatase is associated with the metastatic phenotype in human colorectal tumors.
We have performed differential display comparing gene expression from cell lines derived from human colorectal tumors. The cell lines were selected for study based on their ability to form metastases following injection into athymic mice. One gene which was expressed exclusively by the metastatic lines was identified as human acylphosphatase (e.c. 126.96.36.199, acylphosphate phosphohydrolase). The expression of this gene was confirmed by RT-PCR using gene-specific primers. This gene product catalyzes the hydrolysis of phosphorylated intermediates of Na+/K(+)-ATPase and of Ca(2+)-ATPases of mammalian cells. Changes in the activity of the Na+/K(+)-ATPase pump, regulated by acylphosphatase, have been previously reported in chemically-induced colonic tumors. The differential expression of this gene in the human metastatic colorectal lines suggests it may be involved in the metastatic phenotype.


Alpha-1-antitrypsin phenotypes in adult liver disease patients.
Alpha-1-antitrypsin (AAT) is an important serine protease inhibitor in humans. Hereditary alpha-1-antitrypsin deficiency (AATD) affects lungs and liver. Liver disease caused by AATD in paediatric patients has been previously well documented. However, the association of liver disease with alpha-1-antitrypsin gene polymorphisms in adults is less clear. Therefore, we aimed to study AAT polymorphisms in adults with liver disease. We performed a case-control study. AAT polymorphisms were investigated by isoelectric focusing in 61 patients with liver cirrhosis and 9 patients with hepatocellular carcinoma. The control group consisted of 218 healthy blood donors. A significant deviation of observed and expected frequency of AAT phenotypes from Hardy-Weinberg equilibrium (chi-square = 34.77, df 11, P = 0.000) in the patient group was caused by a higher than expected frequency of Pi ZZ homozygotes (f = 0.0143 and f = 0.0005, respectively, P = 0.000). In addition, Pi M homozygotes were more frequent in patients than in controls (63% and 46%, respectively, P = 0.025). Our study results show that Pi ZZ homozygosity in adults could be associated with severe liver disease. Presence of Pi M homozygosity could be associated with liver disease via some mechanism different from Z allele-induced liver damage through accumulation of AAT polymers.


Association between Regular Aspirin Use and Circulating Markers of Inflammation: A Study within the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial.
Regular aspirin use may decrease cancer risk by reducing chronic inflammation. However, associations between aspirin use and circulating markers of inflammation have not been well studied. Serum levels of 78 inflammatory markers were measured in 1,819 55- to 74-year-old men and women in the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial. Data were combined from three completed case-control studies and reweighted to the PLCO screening arm. Self-reported aspirin and ibuprofen use (number of tablets taken per day/week/month) over the previous 12 months was collected at baseline. Associations between (i) nonregular (<4 tablets/month), (ii) low (1-4 tablets/week), (iii) moderate (1 tablet/day), or (iv) high (2+ tablets/day) regular aspirin or ibuprofen use and marker levels were assessed with weighted logistic regression. Aspirin use was nominally associated with (Ptrend across categories ≤ 0.05) decreased levels of chemokine C-C motif ligand 15 [CCL15; OR, 0.5; 95% confidence intervals (CI), 0.3-0.8; moderate versus nonregular use]; soluble vascular endothelial growth factor receptor 2 (sVEGFR2; OR, 0.7; 95% CI, 0.4-1.0); soluble tumor necrosis factor receptor 1 (sTNFR1; OR, 0.6; 95% CI, 0.4-0.9) and increased levels of CCL13 (OR, 1.3; 95% CI, 0.8-2.1); CCL17 (OR, 1.1; 95% CI, 0.7-1.9) and interleukin 4 (IL4; OR, 1.6; 95% CI, 0.9-2.8). Trends were not statistically significant following correction for multiple comparisons. Likewise, no statistically significant associations were observed between ibuprofen use and marker levels. No significant associations were observed between regular aspirin use and the inflammatory markers assessed. Additional studies are needed to better understand the relationship between aspirin use, chronic inflammation, and cancer risk.


Compartment model predicts VEGF secretion and investigates the effects of VEGF trap in tumor-bearing mice.
Angiogenesis, the formation of new blood vessels from existing vasculature, is important in tumor growth and metastasis. A key regulator of angiogenesis is vascular endothelial growth factor (VEGF), which has been targeted in numerous anti-angiogenic therapies aimed at inhibiting tumor angiogenesis. Systems biology approaches, including computational modeling, are useful for understanding this complex biological process and can aid in the development of novel and effective therapeutics that target the VEGF family of proteins and receptors. We have developed a computational model of VEGF transport and kinetics in the tumor-bearing mouse, which includes three-compartments: normal tissue, blood, and tumor. The model simulates human tumor xenografts and includes human (VEGF121 and VEGF165) and mouse (VEGF120 and VEGF164) isoforms. The model incorporates molecular interactions between these VEGF isoforms and receptors (VEGFR1 and VEGFR2), as well as co-receptors (NRP1 and NRP2). We also include important soluble factors: soluble VEGFR1 (sFlt-1) and α-2-macroglobulin. The model accounts for transport via macromolecular transendothelial permeability, lymphatic flow, and plasma clearance. We have fit the model to available in vivo experimental data on the plasma concentration of free VEGF Trap and VEGF Trap bound to mouse and human VEGF in order to estimate the rates at which parenchymal cells (myocytes and tumor cells) and endothelial cells secrete VEGF. Interestingly, the predicted tumor VEGF secretion rates are significantly lower (0.007-0.023 molecules/cell/s, depending on the tumor microenvironment) than most reported in vitro measurements (0.03-2.65 molecules/cell/s). The optimized model is used to investigate the interstitial and plasma VEGF concentrations and the effect of the VEGF-neutralizing agent, VEGF Trap (aflibercept). This work complements experimental studies performed in mice and provides a framework with which to examine the effects of anti-VEGF agents, aiding in the optimization of such anti-angiogenic therapeutics as well as analysis of clinical data. The model predictions also have implications for biomarker discovery with anti-angiogenic therapies.


Health-related quality of life of people with HIV: an assessment of patient related factors and comparison with other chronic diseases.
The health-related quality of life (HRQOL) of people with HIV is lower than in the general population, but it is unknown how it compares with that of persons with other chronic medical conditions. We compared HRQOL in HIV with HRQOL in diabetes mellitus type 1, diabetes mellitus type 2 and rheumatoid arthritis (RA). In addition, we investigated factors associated with HRQOL in HIV. Cross-sectional study. HRQOL was measured with the Medical Outcomes Study Short Form 36-item Health Survey in a nationwide sample of people with HIV in care in the Netherlands and on combination antiretroviral therapy for at least 6 months. We added data from studies in diabetes mellitus types 1 and 2, and RA. Logistic regression analysis was used to examine: the association between disease group and a poor HRQOL, and patient factors associated with poor HRQOL in HIV. The odds of a poor physical HRQOL in the HIV group were comparable with the odds in diabetes mellitus types 1 and 2, but lower than in RA patients. The odds of a poor mental HRQOL in HIV were higher than in the other groups. In HIV, a history of AIDS, longer duration of combination antiretroviral therapy and severe comorbidity were associated with a poor physical HRQOL. Sub-Saharan African descent and CD4 cell count of less than 350 cells/μl were associated with poor mental HRQOL. People with HIV were more likely to have a poor mental HRQOL than patients with other chronic conditions. Addressing mental health should be an integral part of outpatient HIV care.


Development of criteria to distinguish inflammatory from noninflammatory arthritis, enthesitis, dactylitis, and spondylitis: a report from the GRAPPA 2013 Annual Meeting.
To describe a research project to develop simple clinical criteria to aid in the identification of inflammatory arthritis, enthesitis, dactylitis, and spondylitis and distinguish these from non-inflammatory conditions. The criteria are particularly intended to aid non-rheumatologists, e.g., dermatologists, who need assistance identifying psoriatic arthritis in patients with psoriasis, but may be useful to all clinicians in properly diagnosing rheumatologic conditions. The proposed research methodology includes the use of a nominal group exercise among expert clinicians and patient focus groups, Delphi exercises among clinicians and patients, application of criteria test sets to a small group of representative patients with inflammatory and non-inflammatory musculoskeletal conditions, and validation by application of optimal criteria sets to large groups of patients with inflammatory and noninflammatory conditions. Examples of elements to describe inflammatory conditions derived from a nominal group exercise conducted at the 2013 GRAPPA annual meeting are described, along with planned project activities. This project will lead to the development of practical criteria to aid in the diagnosis and appropriate clinical care of patients with chronic inflammatory musculoskeletal conditions.


Pathogenesis of myocardial injury in myocarditis and cardiomyopathy.
The pathogenesis of myocardial cell injury in myocarditis and cardiomyopathy was investigated. The presence of viral genomes in the murine heart in experimental coxsackievirus B3 myocarditis was studied by Northern blotting analysis using a 32P-labeled cDNA probe from the 5' end sequence. The strongest signal of positive autoradiograms was always at about 7.4 kilobases, corresponding to the size of the complete genome of the virus. Successive infections with coxsackievirus and encephalomyocarditis (EMC) virus infection showed simultaneous acute myocarditis and healed myocarditis, and the results suggest that successive virus infections cause additional myocardial damage, and develop lesions similar to chronic myocarditis or dilated cardiomyopathy. Anti-heart auto-antibody, induced during EMC virus infection, reacted predominantly with myosin. Indium-111 antimyosin scintigraphy showed positive in some of the patients with cardiomyopathy, and the uptake was inversely correlated with left ventricular function. When mice were injected with antimyosin antibody, mouse immunoglobulin G was detected in hearts in the chronic stage of EMC virus myocarditis, in myocytes surrounding fibrosis and calcification, suggesting deposition of antimyosin antibody. Although further study is necessary to clarify the mechanism of uptake of antimyosin in cardiomyopathy, antimyosin antibody may accumulate in viable myocytes with ongoing degeneration as well as in necrosis.


Effects of Cataract Opacity and Surgery on Sleep Quality.
This study aimed to identify the type of cataract opacity associated with subjective sleep quality for cataract patients. A total of 180 consecutive patients (average age 74.2 years) underwent cataract surgery with implantation of an ultraviolet-blocking or blue light-blocking intraocular lens. The participants' subjective sleep quality was evaluated using the Pittsburgh sleep quality index (PSQI) before and at 2 months after surgery. Patients were divided into two groups for analysis: normal sleepers (preoperative PSQI ≤5) and poor sleepers (preoperative PSQI ≥6). The preoperative and postoperative PSQI scores were 2.8 ± 1.5 and 3.2 ± 2.0, respectively, for normal sleepers (n = 99), and 8.5 ± 2.9 and 7.4 ± 3.3, respectively, for poor sleepers (n = 81). The improvement in PSQI was significant in poor sleepers (p < 0.0001, Wilcoxon's test) with measured values of 0.22 ± 0.96 hours extension in sleep duration and 0.16 ± 0.62 hours shortening in sleep latency. Patients with posterior subcapsular cataract (PSC) and nuclear opacity showed the greatest improvement in PSQI score, with regression analysis identifying PSC as having a significant effect on improvement in PSQI. Significant correlations were found between the subjective sleep and cataract opacity in cataract patients. Ophthalmic personnel should therefore be aware that cataract patients with PSC have a greater potential for disability and predictable benefits from surgery in vision and subjective sleep.


Changes in CD4+ T-cell differentiation phenotype during structured treatment interruption in patients with chronic HIV-1 infection.
Markers of maturation and activation were measured on peripheral CD4+ T cells in chronically HIV-1-infected patients in a randomized, controlled pilot study of structured treatment interruption (STI). Eight subjects underwent 2 cycles of 1 month off and 1 month on highly active antiretroviral therapy (HAART), followed by a final 3-month interruption. During STI, CD4+ T-cell percentage remained relatively stable in 4 of 8 subjects. The remaining 4 STI subjects had significant rapid decline in CD4+ T-cell percentage during STI, followed by return to pre-STI baseline while on HAART. Changes in overall CD4+ T-cell percentage corresponded with fluctuations in the CD45RA+CCR7+ naive and CD45RA-CCR7+ central memory subsets. Subjects with variable CD4+ T-cell percentages tended to have higher pre-HAART plasma HIV-1 RNA set-points and experienced higher levels of plasma HIV-1 RNA rebound during STI. These results suggest that interruptions should be avoided whenever possible in patients on HAART with high plasma HIV-1 RNA set-points.


Immunotherapy trials for type 1 diabetes: the contribution of George Eisenbarth.
Type 1 diabetes (T1D) results from the autoimmune destruction of pancreatic β-cells, and as such it should respond to immunotherapy. George Eisenbarth gave many significant contributions to this field. He has been involved at some level in most immunotherapy trials during the past three decades. He was among the pioneers who attempted immunotherapy approaches in patients with recent-onset T1D. In the early 1980s he began studying relatives of those with the disease, leading to the concept that T1D was a chronic autoimmune disease, in which islet autoimmune responses would silently destroy β-cells and cause progressive impairment of insulin secretion, years to months before a diagnosis was made. Consequently, he was one of the first to attempt immune intervention in people at high risk of T1D. Throughout his career he developed autoantibody assays and predictive models (which included metabolic testing and later genetics) to identify individuals at risk of T1D. He provided seminal intellectual contributions and critical tools for prevention trials. His focus on insulin as a critical autoantigen led to multiple prevention trials, including the Diabetes Prevention Trial-Type 1 (DPT-1), which studied both parenteral and oral insulin. In the DPT-1 Oral Insulin Trial, a cohort with higher levels of insulin autoantibodies was identified that appeared to have delayed disease progression. Type 1 Diabetes TrialNet is conducting a new trial to verify or refute this observation. Moreover, George identified and tested in the mouse small molecules that block or modulate presentation of a key insulin peptide and in turn prevent the activation of insulin-specific T-lymphocytes. Thus, we believe his greatest contribution is yet to come, as in the near future we should see this most recent work translate into clinical trials.


Effect of zinc supplementation on the status of thyroid hormones and Na, K, And Ca levels in blood following ethanol feeding.
The influence of zinc (Zn) on the serum levels of triiodothyronine (T(3)), thyroxine (T(4)), thyroid-stimulating hormone (TSH) and sodium (Na), potassium (K), and calcium (Ca) was evaluated following ethanol toxicity to the rats. To achieve this, male Wistar rats (150-195 g) were given 3 ml of 30% ethanol orally, and zinc was given in the form of zinc sulfate (227 mg/l) in their drinking water daily for 8 weeks. Ethanol feeding resulted in a slight decrease in T(3) and T(4) levels and a significant increase in thyroid-stimulating hormone concentration, which may be due to the direct stimulatory effect of ethanol on thyroid. Interestingly, when zinc was given to these rats, all the above levels were brought quite close to their normal levels, thus indicating the positive role of zinc in thyroid hormone metabolism. Serum Zn and Ca levels were found to be reduced, but Na levels were raised upon ethanol feeding. Restoration of normal levels of these metals upon zinc supplementation to ethanol fed rats confirms that zinc has potential in alleviating some of the altered thyroid functions following ethanol administration.


The universal properties of stem cells as pinpointed by a simple discrete model.
The ability of a few stem-cells to repopulate a severely damaged bone marrow (BM) guarantees the stability of our physical existence, and facilitates successful BM transplantations. What are the basic properties of stem cells that enable the maintenance of the system's homeostasis? In the present work we attempt to answer this question by investigating a discrete (in time and phase-space) dynamical system. The model we present is shown to retrieve the essential properties of homeostasis, as reflected in BM functioning, namely, (a) to produce a constant amount of mature cells, and (b) to be capable of returning to this production after very large perturbations. The mechanism guaranteeing the fulfillment of these properties is extrinsic--negative feedback control in the micro-environment--and does not need additional stochastic assumptions. Nevertheless, the existence of a simple intrinsic control mechanism, a clock which determines the switch to differentiation, ascertains that the system does not admit non-trivial extinction states. This result may help clarifying some of the controversy about extrinsic versus intrinsic control over stem cell fate. It should be stressed that all conclusions are valid for any system containing progenitor and maturing cells.


Patterns of Wnt/Fzd/LRP gene expression during embryonic hematopoiesis.
Wnt signaling plays several roles in hematopoiesis, promoting hemopoietic stem cell (HSC) self-renewal, providing proliferative signals for immature progenitors and regulating lineage commitment. To ascertain which Wnt proteins and receptors are important during hematopoietic development, we used two systems; in vitro hematopoietic differentiation of embryonic stem (ES) cells and tissues isolated from sites specific for hematopoiesis during mouse embryogenesis. Initially genes involved in hematopoiesis were profiled and indicate differentiating ES cells undergo a wave of primitive hematopoiesis (Day 3.75) similar to the mouse yolk sac, followed by a wave of more definitive hematopoiesis (Day 7.75) comparable to the aorta-gonad-mesonephros (AGM) and E15.5 liver with lineage commitment by Day 15. A similar biphasic expression pattern occurred for Wnt/Fzd/LRP genes with Wnt 3, 5a, 8a, Fzd4, and LRP5 becoming upregulated during primitive hematopoiesis, followed by Wnt3a, 6, 7b, 10b, and 16 during more definitive hematopoiesis. High expression of Wnt5a, Fzd4, and LRP5 during the first phase of hematopoiesis suggests these genes are involved in early hematopoietic regulation. Wnt3a and 16 were also expressed at specific stages, with Wnt16 detected when the earliest lymphoid progenitors are formed (AGM and 2 degrees BC of ES differentiation). Wnt3a expression corresponded with the induction of definitive hematopoiesis a period, which involves rapid expansion of HSC (Day 7.75 of ES differentiation, AGM and E15.5 liver). Supplementation with Wnt3a during ES hematopoietic differentiation increased proliferation and appeared to promote stem cell expansion. Overall this study provides valuable information on the Wnt/Fzd/LRP involved in supporting embryonic hematopoiesis.


Paracrine and autocrine regulation of gene expression by Wnt-inhibitor Dickkopf in wild-type and mutant hepatocytes.
Cells are able to communicate and coordinate their function within tissues via secreted factors. Aberrant secretion by cancer cells can modulate this intercellular communication, in particular in highly organised tissues such as the liver. Hepatocytes, the major cell type of the liver, secrete Dickkopf (Dkk), which inhibits Wnt/ β-catenin signalling in an autocrine and paracrine manner. Consequently, Dkk modulates the expression of Wnt/ β-catenin target genes. We present a mathematical model that describes the autocrine and paracrine regulation of hepatic gene expression by Dkk under wild-type conditions as well as in the presence of mutant cells. Our spatial model describes the competition of Dkk and Wnt at receptor level, intra-cellular Wnt/ β-catenin signalling, and the regulation of target gene expression for 21 individual hepatocytes. Autocrine and paracrine regulation is mediated through a feedback mechanism via Dkk and Dkk diffusion along the porto-central axis. Along this axis an APC concentration gradient is modelled as experimentally detected in liver. Simulations of mutant cells demonstrate that already a single mutant cell increases overall Dkk concentration. The influence of the mutant cell on gene expression of surrounding wild-type hepatocytes is limited in magnitude and restricted to hepatocytes in close proximity. To explore the underlying molecular mechanisms, we perform a comprehensive analysis of the model parameters such as diffusion coefficient, mutation strength and feedback strength. Our simulations show that Dkk concentration is elevated in the presence of a mutant cell. However, the impact of these elevated Dkk levels on wild-type hepatocytes is confined in space and magnitude. The combination of inter- and intracellular processes, such as Dkk feedback, diffusion and Wnt/ β-catenin signal transduction, allow wild-type hepatocytes to largely maintain their gene expression.


Cytogenetics of collared lemmings (Dicrostonyx groenlandicus). I. Meiotic behavior and evolution of the neo-XY sex-chromosome system.
Electron-microscopic analysis of surface-spread synaptonemal complexes at pachynema and light-microscopic analysis of chromosomal configurations at diakinesis/metaphase I corroborate the hypothesized neo-XY derivation of the sex chromosomes of Dicrostonyx groenlandicus. Although an intact neo-XY pairing configuration was observed in a relatively small percentage of the pachytene cells in each individual, the high incidence of neo-XY bivalents at diakinesis/metaphase I suggests that the other observed pachytene configurations were artifacts of the physical stresses of the surface-spreading procedure. The very low frequency (0.6%) of univalent neo-X and neo-Y chromosomes at diakinesis and metaphase I is attributable to consistent synapsis and recombination between their homologous autosomally derived segments. The resultant stability of the sex bivalent through metaphase I may have increased the efficacy of sex-chromosome segregation, and thereby played a mechanistic role in the evolutionary incorporation of the neo-XY sex-chromosome constitution in D. groenlandicus.


Antiemetic studies: a methodological discussion.
Methodological issues encountered in reviewing the antiemetic literature are discussed. Variables affecting antiemetic assessment include those relating to the patient, the emetic stimulus, and the differences in action of the antiemetic drugs. Study design questions including the desirability of randomization, double blinding, use of controls, and stratifications for factors such as previous exposure to chemotherapy and strength of the emetic stimulus are discussed. From a statistical viewpoint a parallel subject design with adequate sample size is recommended. The problems of standardizing the evaluation of nausea and vomiting are considered and simple ordinal scales grading severity are suggested. The patient's overall assessment of tolerance of therapy is of primary importance.


COVER: a priori estimation of coverage for metagenomic sequencing.
In any metagenomic project, the coverage obtained for each particular species depends on its abundance. This makes it difficult to determine a priori the amount of DNA sequencing necessary to obtain a high coverage for the dominant genomes in an environment. To aid the design of metagenomic sequencing projects, we have developed COVER, a web-based tool that allows the estimation of the coverage achieved for each species in an environmental sample. COVER uses a set of 16S rRNA sequences to produce an estimate of the number of operational taxonomic units (OTUs) in the sample, provides a taxonomic assignment for them, estimates their genome sizes and, most critically, corrects for the number of unobserved OTUs. COVER then calculates the amount of sequencing needed to achieve a given goal. Our tests and simulations indicate that the results obtained through COVER are in very good agreement with the experimental results.


Rapid detection of extra-intestinal pathogenic Escherichia coli multi-locus sequence type 127 using a specific PCR assay.
Members of the ST127 uropathogenic E. coli (UPEC) clone have a high virulence potential and are also highly virulent in insect infection models. However, strains of this lineage are reported in relatively low numbers in many studies. ST127 strains are also usually widely susceptible to antibiotics and, consequently, their true prevalence may be under-recognized as they will be eradicated during empirical therapy. A genuine concern is the possibility that members of this highly virulent lineage will acquire resistance, leading to a more serious threat. The aim of this study was to design and validate a PCR assay specific to ST127. Genomic sequences obtained from various UPEC isolates from the leading clones were used in comparative genomic analyses to allow identification of highly discriminatory sequences specific to E. coli ST127. The fliC (flagellin) and a homologue of the upaG (autotransporter adhesin) gene were identified as meeting our criteria and were used to develop a multiplex PCR assay. A total of 143 UPEC isolates representing 99 different MLST clones from three locations (North West and South West England and Riyadh, Saudi Arabia) were used to validate the PCR assay. The multiplex PCR readily identified all 29 E. coli ST127 isolates but, equally importantly, produced no false positives with representatives of any of the other 98 STs tested. We report the design and validation of a specific multiplex PCR for the rapid and reliable identification of ST127, which can be used for enhanced surveillance for this high-risk clone.


FBXO31 determines poor prognosis in esophageal squamous cell carcinoma.
Cyclin D1 plays important roles in esophageal squamous cell carcinoma (ESCC) cases by amplification of the 11q13.3 locus. FBXO31 is a subunit of the SCF ubiquitin ligase, which targets cyclin D1 for degradation. In this study, we clarified the clinical significance of FBXO31 and characterized the association between cyclin D1 and FBXO31 in ESCC cases. Total RNA was extracted from tumor tissues obtained from 68 ESCC patients who underwent surgical resection. FBXO31 expression levels were determined by quantitative RT-PCR, and both FBXO31 and cyclin D1 protein expression and localization were evaluated by immunohistochemistry (IHC). Furthermore, using CGH and gene expression array data of another subset, we validated the association between cyclin D1 genomic amplification and FBXO31 expression levels. Higher FBXO31 expression levels significantly correlated with depth of tumor invasion and clinical stage (P<0.05). In addition, the FBXO31 high expression group showed a significantly poorer prognosis than the low expression group (P<0.001). Multivariate analysis indicated that FBXO31 expression was an independent prognostic factor [relative risk (RR): 1.79, confidence interval (CI): 1.14-3.01, P=0.01]. Using IHC, concordant expression was observed between cyclin D1 and FBXO31 in the nucleus and cytoplasm, respectively. CGH array indicated that cases having cyclin D1 with increased copy number were significantly associated with elevated FBXO31 expression levels (P<0.05). FBXO31 could be a novel and robust prognostic marker for ESCC.


Cytotoxic and cytokinetic effects of 1-beta-D-arabinofuranosylcytosine, daunorubicin, and 6-thioguanine on HeLa cells in culture.
Exponentially growing HeLa cells were exposed to 1-beta-D-arabinofuranosylcytosine, daunorubicin, and 6-thioguanine singly and in combination for 21 hr. Survivals were determined by a colony-counting assay, and cytokinetic effects of the drugs were evaluated by flow cytometry. When used in two-drug combinations (concurrent exposures). 1-beta-D-arabinofuranosylcytosine was found to protect cells against both 6-thioguanine- and daunorubicin-induced lethality, the former to a greater extent than the latter. Protection against 6-thioguanine appeared to be related to the degree of inhibition of DNA synthesis by 1-beta-D-arabinofuranosylcytosine, but this was not the case with daunorubicin. When cells were exposed to the three drugs simultaneously, survivals were in agreement with those predicted from the various two-drug combinations. This implies that no additional drug interactions occurred when a third drug was added to the two-drug combinations.


Chromosomal anomalies in stage D1 prostate adenocarcinoma primary tumors and lymph node metastases detected by fluorescence in situ hybridization.
We determined if characteristic chromosomal anomalies exist within the primary tumors and lymph node metastases in patients with stage D1 prostate cancer, and compared the patterns of chromosomal alterations between primary tumors and nodal metastases. Fluorescence in situ hybridization analysis using peri-centromeric probes for chromosomes 6, 7, 8, 17, X and Y was performed on 5 mu. sections from paraffin embedded tissue blocks obtained from 23 consecutive patients who underwent radical prostatectomy and bilateral pelvic lymphadenectomy in 1990 for stage D1 prostate cancer. The dominant focus of primary tumor was compared to matched nodal metastases in 12 cases. Five of 12 primary tumor foci (41.7%) had similar chromosomal gains and the same fluorescence in situ hybridization ploidy result as the corresponding nodal metastases. Chromosomes 7 and X (73.2% of cases) were most frequently gained in the primary tumors, and chromosomes X and Y (81.2% of cases) were most frequently gained in the metastases. No primary tumor or metastasis demonstrated chromosomal loss. Three of 19 primary tumors (15.7%) were diploid, while 16 of 19 (84.3%) were nondiploid. Chromosomal aneusomy was inversely correlated with increasing Gleason summary score. These data indicate that the dominant primary tumor foci may not give rise to nodal metastases, gains of chromosomes 7, X and Y may be associated with metastatic behavior, and patients with stage D1 disease have a greater rate of aneuploidy than those with lower stage cancer.


Early and late onset complications of gynaecologic surgery: a multimodality imaging approach.
The role of imaging after surgery is pivotal to drive clinical management of early and/or late onset complications. Most frequently used imaging technique after pelvic surgery is Ultrasound (US), Magnetic Resonance Imaging (MRI) and Computed Tomography (CT). While Ultrasound is a standard procedure, using grey scale and/or colour Doppler evaluation, MRI and CT scan protocols should be derived on the basis of the specific indication of the exam. Correct evaluation of female pelvis after gynaecologic surgery, having in mind the most frequent complications, is based on the correct use of the instruments and on the experience of the examiner, who should be aware of the history of the patient, type of surgery and clinical symptoms for which the exam is required; the clinician should be aware of the possibilities and limits of the different techniques, in order to choose the most appropriate imaging modality and promptly make a correct diagnosis.


Detailed Analyses of Zika Virus Tropism in Culex quinquefasciatus Reveal Systemic Refractoriness.
The role of Culex quinquefasciatus in Zika virus transmission has been debated since the epidemic of Zika occurred in the Americas in 2015 to 2016. The majority of studies have found no evidence that C. quinquefasciatus or other Culex species are competent vectors of Zika virus, and the few studies that have proposed Zika vector status for C. quinquefasciatus have relied predominantly on quantitative real-time PCR (qRT-PCR) for viral detection. We assessed the infectious range of pre- and post-epidemic Zika virus isolates in order to classify mosquito samples based on titer infectiousness and demonstrated that two strains of C. quinquefasciatus, including one previously found to be competent, are highly resistant to infection with these Zika isolates compared to Aedes aegypti and are not competent for virus transmission. Further dissection of the dynamics of Zika exposure in both A. aegypti and C. quinquefasciatus revealed that while virus transmission by C. quinquefasciatus is blocked at the levels of the midgut and salivary glands, viral RNA persists in these tissues for prolonged periods post-exposure. We assessed Zika entry dynamics in both Aedes and Culex cells, and our results suggest that Zika virus infection in Culex cells may be blocked downstream of cell entry. These findings strongly suggest that C. quinquefasciatus is not a vector of Zika virus and additionally inform the use of qRT-PCR in vector competence assays as well as our understanding of barriers to arbovirus infection in non-susceptible mosquito species.IMPORTANCE Understanding which mosquito species transmit an emerging arbovirus is critical to effective vector control. During the Zika virus epidemic in 2015 to 2016, Aedes mosquitoes were confirmed as vectors. However, studies addressing the vector status of Culex quinquefasciatus mosquitoes presented conflicting evidence and remain an outstanding source of confusion in the field. Here, we established a robust cell-based assay to identify infectious titers of Zika virus and assessed the virus titers in C. quinquefasciatus by quantitative real-time PCR (qRT-PCR). We found that while low levels of virus were detected in C. quinquefasciatus, these titers did not correspond to infectious virus, and these mosquitoes did not transmit virus in the saliva. We also present evidence that the virus may enter Culex cells before infection is disrupted. Our findings are important for future studies incriminating vector species using qRT-PCR for virus detection and offer new information on how virus transmission is blocked by mosquitoes.