Breast cancer in Bedouin-Arab patients in southern Israel: epidemiologic and biologic features in comparison with Jewish patients.
Breast cancer (BC) is the most frequent cancer type, and the leading cause of death from cancer among women in Israel. The Bedouin-Arab (BA) population in southern Israel is characterized by a high rate of consanguinity, common hereditary disorders, and transition from a semi-nomadic, traditional society to a more sedentary and urbanized society. In this hospital-based study, the demographic and the clinicopathological characteristics of BC in BA were compared with Jewish patients. 85 BA patients treated at the Soroka Medical Center, Beer Sheba, during the years 2004-2012, were studied and compared with 180 consecutive Jewish patients treated during the year 2007. Clinicopathological features compared included age, menopausal state, number of births, a history of BC in first-degree relatives, tumor size (T), extent of lymph- node involvement (N), distant metastases (M), stage, grade, estrogen and progesterone receptor (ER/PR), and Her2 status. Types of treatment, relapse rate and site, as well as outcome were also studied. Cox's regression models were applied for studying disease-free, and overall survival. Compared with Jewish patients, BA patients were younger (average age 49±12 yrs vs 59±13, p<0.001), had a lower rate of BC in first-degree relatives (p<0.001), and a larger number of births (6±4.2 vs 2.5±1.9, p<0.001). BA patients had larger tumors (p=0.02), more extensive lymph-node involvement (p=0.002), and more advanced stage (p=0.003). Grade, ER, PR, and Her2 status were similar in the two ethnic groups. Relapse type was most commonly systemic in BA patients (p=0.05), and loco-regional in Jewish patients (p=0.02). Median survival was 63, and 35 months for Jewish and BA patients, respectively (log-rank test, p=0.02). In Cox multivariate analysis, stage and PR status (HR-0.14, p<0.0001; HR-3.11, p=0.046), but not ethnicity, influenced overall survival. BC presents a decade earlier, and with more advanced disease in BA compared with Jewish patients. Biologic parameters including grade, ER, PR, and Her2 status were similar in both groups. Although prognosis was worse in BA than in Jewish patients, it was affected only by stage and PR status, but not by ethnicity.


Retinoic acid. Inhibition of the clonal growth of human myeloid leukemia cells.
Vitamin A and its analogues (retinoids) affect normal and malignant hematopoietic cells. We examined the effect of retinoids on the clonal growth in vitro of myeloid leukemia cells. Retinoic acid inhibited the clonal growth of the KG-1, acute myeloblastic leukemia, and the HL-60, acute promyelocytic leukemia, human cell lines. The KG-1 cells were extremely sensitive to retinoic acid, with 50% of the colonies inhibited by 2.4-nM concentrations of the drug. A 50% growth inhibition of HL-60 was achieved by 25 nM retinoic acid. Complete inhibition of growth of both leukemia cell lines was seen with 1 microM retinoic acid. Exposure of KG-1 cells to retinoic acid for only 3-5 d was sufficient to inhibit all clonal growth. The all-trans and 13-cis forms of retinoic acid were equally effective in inhibiting proliferation. Retinal, retinyl acetate, and retinol (vitamin A) were less potent inhibitors. Clonal growth of the human K562 and mouse M-1 myeloid leukemic cell lines was not affected by 10 microM retinoic acid. Retinoic acid also inhibited the clonal growth of leukemia cells from five of seven patients with acute myeloid leukemia. Retinoic acid at concentrations of 5 nM to 0.3 microM inhibited 50% clonal growth, and 1 microM retinoic acid inhibited 64-98% of the leukemic colonies. The inhibition of clonal growth of KG-1 and HL-60 cell lines and of leukemic cells from two patients was not associated with the presence of a specific cytoplasmic retinoic acid-binding protein. Our study suggests that retinoic acid may prove to be effective in the treatment of human myeloid leukemia.


Treatment of snoring and obstructive sleep apnea with mandibular repositioning appliances.
Snoring and obstructive sleep apnea form part of a spectrum of sleep disordered breathing affecting a significant proportion of the general population and particularly the middle aged. The consequences can be severe and even life threatening for both the individual directly affected and those more remotely involved. Adverse sequelae can manifest themselves acutely or in the longer term as a result of obstructive breathing induced hypersomnolence, neurocognitive deficits and cardiovascular abnormalities. The combination of anatomical and neuromuscular risk factors in the pathogenesis of OSA has resulted in a varied approach to its management. One such treatment option is mandibular repositioning appliances (MRA), which mechanically stabilize the airway. Whilst the efficacy of this simple intervention has been rigorously proven quite recently in a significant proportion of patients with varying disease severity, individual patient selection in its application remains uncertain. Short-term side-effects are common but usually transient, whilst in the long-term minor permanent adverse developments on the dentition and occlusion have been reported. Considering both the medicolegal implications of snoring and OSA and the increasing popularity of MRA, it is recommended that skilled multidisciplinary respiratory and dental personnel form the primary care team.


Supplemental levels of iron and calcium interfere with repletion of zinc status in zinc-deficient animals.
Negative interactions between minerals interfering with each other's absorption are of concern when iron and calcium supplements are given to pregnant women and children. We have previously reported that supplemental levels of iron and calcium inhibit the bioaccessibility of zinc, and compromise zinc status in rats fed diets with high levels of these two minerals. The present study examined the effect of supplemental levels of iron and calcium on the recovery of zinc status during a zinc repletion period in rats rendered zinc-deficient. Iron and calcium, both individually and in combination, significantly interfered with the recovery of zinc status in zinc deficient rats during repletion with normal levels of zinc in the diet. Rats maintained on diets containing supplemental levels of these two minerals had significantly lower body weight, and the concentration of zinc in serum and organs was significantly lower than in zinc-deficient rats not receiving the supplements. Iron and calcium supplementation also significantly inhibited the activity of zinc-containing enzymes in the serum as well as liver. Both iron and calcium independently exerted this negative effect on zinc status, while their combination seemed to have a more prominent effect, especially on the activities of zinc containing enzymes. This investigation is probably the first systematic study on the effect of these two minerals on the zinc status of zinc deficient animals and their recovery during repletion with normal amounts of zinc.


Management of lymphatic fistulas by total parenteral nutrition.
Lymphatic fistula is a rare surgical complication, which mainly occurs after cervical or retroperitoneal lymph node dissection and which frequently requires a surgical repair. A small series of nine postoperative lymphatic fistulas treated conservatively with total parenteral nutrition (TPN) is reported. All the patients were malnourished at the beginning of the TPN, and all exhibited an objective improvement of their nutritional status after completion of the treatment. Due to the interruption of the enteral alimentation or to the nutritional repletion, spontaneous closure of the fistula was achieved in eight of nine patients treated with TPN longer than 1 week. The authors conclude that whenever the immediate surgical repair is not recommended, or it is not successful, a 2- to 3-week course of TPN may be used with the chance of spontaneous healing. In any case, patients conservatively treated by TPN can undergo a delayed operation with minimal risk because of the improved nutritional status.


Delays in appropriate antibiotic therapy for gram-negative bloodstream infections: a multicenter, community hospital study.
Gram-negative bacterial bloodstream infection (BSI) is a serious condition with estimated 30% mortality. Clinical outcomes for patients with severe infections improve when antibiotics are appropriately chosen and given early. The objective of this study was to estimate the association of prior healthcare exposure on time to appropriate antibiotic therapy in patients with gram-negative BSI. We performed a multicenter cohort study of adult, hospitalized patients with gram-negative BSI using time to event analysis in nine community hospitals from 2003-2006. Event time was defined as the first administration of an antibiotic with in vitro activity against the infecting organism. Healthcare exposure status was categorized as community-acquired, healthcare-associated, or hospital-acquired. Time to appropriate therapy among groups of patients with differing healthcare exposure status was assessed using Kaplan-Meier analyses and multivariate Cox proportional hazards models. The cohort included 578 patients with gram-negative BSI, including 320 (55%) healthcare-associated, 217 (38%) community-acquired, and 41 (7%) hospital-acquired infections. 529 (92%) patients received an appropriate antibiotic during their hospitalization. Time to appropriate therapy was significantly different among the groups of healthcare exposure status (log-rank p=0.02). Time to first antibiotic administration regardless of drug appropriateness was not different between groups (p=0.3). The unadjusted hazard ratios (HR) (95% confidence interval) were 0.80 (0.65-0.98) for healthcare-associated and 0.72 (0.63-0.82) for hospital-acquired, relative to patients with community-acquired BSI. In multivariable analysis, interaction was found between the main effect and baseline Charlson comorbidity index. When Charlson index was 3, adjusted HRs were 0.66 (0.48-0.92) for healthcare-associated and 0.57 (0.44-0.75) for hospital-acquired, relative to patients with community-acquired infections. Patients with healthcare-associated or hospital-acquired BSI experienced delays in receipt of appropriate antibiotics for gram-negative BSI compared to patients with community-acquired BSI. This difference was not due to delayed initiation of antibiotic therapy, but due to the inappropriate choice of antibiotic.


[Fluoroscopic-guided primary single-step percutaneous gastrostomy: initial results using the Freka® GastroTube].
To determine the practicability and outcome of fluoroscopic-guided primary one-step treatment of percutaneous gastrostomy (PG) with the system Freka® Gastro Tube (Fresenius Kabi, Germany). In 39 patients (mean age 62.7 ± 12.0 years), primary PG was performed based on clinical indication from August 2009 to April 2010. The intervention was performed by an experienced radiologist under aseptic conditions by direct puncture with Freka® Gastro Tube under fluoroscopic guidance. The clinical data and outcome as well as any complications originated from the electronic archive of the University Medical Center Hamburg-Eppendorf. The intervention was technically successful in all 39 patients. Within the mean follow-up time of 155.3 ± 73.6 days, 29 patients (74.4 %) did not experience complications. 10 patients (25.6 %) had to be revised. Complications manifested after a mean of 135.6 ± 61.2 days and mainly corresponded to accidental dislocation (50 %). One patient had to be surgically revised under suspicion of a malpositioned tube and suspected intestinal perforation. Clinically relevant wound infections were not detected. The total costs per patient were 553.17 € for our single-step treatment (OPS 5 - 431.x) vs. 963.69 € (OPS 5 - 431.x and OPS 8 - 123.0) for the recommended two-step treatment. Fluoroscopic-guided primary single-step treatment with Freka® Gastro Tube system is feasible and not associated with an increased complication rate when compared to published literature applying a two-step treatment approach. Material costs as well as human and time resources could be significantly reduced using the single-step treatment.


Screening for highly heterozygous chickens in outbred commercial broiler lines to increase detection power for mapping quantitative trait loci.
Two commercial broiler lines were used in an experiment to map quantitative trait loci (QTL) affecting disease resistance. Chickens from these lines were genotyped with 27 microsatellite markers to estimate heterozygosity and polymorphism information content (PIC), the probability that one parent is heterozygous at a marker locus and the other has a different genotype. Heterozygosity estimated from allelic frequencies was 0.52 for the two lines; however, heterozygosity calculated from actual counts of heterozygous loci was much lower (0.36) than the estimated heterozygosity. The PIC was 0.45 in these lines, and average allele number per marker locus was about 3.5. Twenty-five males produced from a cross between these two lines were screened with the DNA markers to select birds with high heterozygosity at marker loci. Mating simulation showed that uninformative matings could be reduced by about 5% if 12 pairs of males and females with the highest heterozygosity at marker loci were selected, which was about a 25% reduction in total uninformative matings. This experiment demonstrated that the heterozygosity and PIC in commercial broiler lines were low and selection for the birds with high heterozygosity at marker loci could increase informative content in chickens used in the experiments of QTL mapping, thus increasing detection power for QTL mapping.


Effect of compounds of the urea-guanidinium class on renaturation and thermal stability of acid-soluble collagen.
The effects of guanidinium salts in decreasing the renaturation rate and lowering the thermal stability of acid-soluble calf-skin collagen have been compared with those of formamide and urea. With the exception of guanidinium sulphate at higher concentrations, no qualitative differences were apparent in the effects of these perturbants, which thus differed only in molar activity. Activity variation in the guanidinium salts reflected a net effect resulting from additivity of cation and anion contributions. As observed in other protein systems, lyotropic activity increased in the series formamide<urea<guanidinium ion, and in the guanidinium salts in the anion order fluoride<sulphate<chloride<bromide<nitrate<iodide. Low activities of guanidinium fluoride and sulphate were attributable to counter-effects of the anions, which acted as structural stabilizers. Changes in renaturation kinetics induced by either temperature or added perturbants appeared to conform with the Flory-Weaver model for the collagen transition. Additivity and non-specificity of the observed effects are discussed with particular reference to a common mechanism involving weak, non-saturated binding of perturbants at protein peptide groups.


Ab initio studies of the optoelectronic structure of undoped and doped silicon nanocrystals and nanowires: the role of size, passivation, symmetry and phase.
Silicon nanocrystals and nanowires have been extensively studied because of their novel properties and their applications in electronic, optoelectronic, photovoltaic, thermoelectric and biological devices. Here we discuss results from ab initio calculations for undoped and doped Si nanocrystals and nanowires, showing how theory can aid and improve comprehension of the structural, electronic and optical properties of these systems.


Secondary Bubble Entrainment via Primary Bubble Bursting at a Viscoelastic Surface.
Bubble bursting at liquid surfaces is ubiquitous and plays a key role for the mass transfer across interfaces, impacting global climate and human health. Here, we document an unexpected phenomenon that when a bubble bursts at a viscoelastic surface of a bovine serum albumin solution, a secondary (daughter) bubble is entrapped with no subsequent jet drop ejection, contrary to the counterpart experimentally observed at a Newtonian surface. We show that the strong surface dilatational elastic stress from the viscoelastic surface retards the cavity collapse and efficiently damps out the precursor waves, thus facilitating the dominant wave focusing above the cavity nadir. The onset of daughter bubble entrainment is well predicted by an interfacial elastocapillary number comparing the effects of surface dilatational elasticity and surface tension. Our Letter highlights the important role of surface rheology on free surface flows and may find important implications in bubble dynamics with a contaminated interface exhibiting complex surface rheology.


[Macroscopic and histologic analysis of the rat prostate after denervation].
To determine the role of autonomic innervation in the preservation of the prostate gland structure, we analyzed the gross and histological features of the rat prostate submitted to denervation by resection of the pelvic ganglia. The study comprised 80 male Sprague-Dawley rats aged 3 months; 42 were anesthetized and resection of the right pelvic ganglia was performed via laparotomy. Thirty days thereafter they were sacrificed. Gross and microscopic analyses of the right ventral lobe were performed. Computerized morphometric analyses of the areas occupied by the lumen, epithelial and stromal compartments, the height of the epithelium and the vascular pattern were performed. The results were compared with those obtained from the 38 ventral lobes of rats of the same age that had not undergone denervation. Gross examination of denervated rat ventral lobe demonstrated atrophy and weighted 36% less (p < 0.001). Histological analysis of the compartments showed 27.9% were epithelial, 48.3% stromal and 51.8% glandular--epithelial and lumen--for the control group versus 14.8% (p < 0.001), 55.7% and 44.4% (not significant), respectively, for the denervated group. Denervated rat epithelial height measured 39.7% less. No differences in the vascular pattern were found. Resection of the rat pelvic ganglia, thereby inducing near-total vesico-prostatic denervation, causes structural and functional changes in the rat prostate compatible with atrophy. The observed changes were a reduction in weight and volume, as well as in the height and surface area of the secretory epithelium.


Identification and cloning potentially protective antigens of Coxiella burnetii using sera from mice experimentally infected with Nine Mile phase I.
Coxiella burnetii is an obligate intracellular bacterium that causes acute Q fever and occasional chronic infections in humans. To determine the immunodominant antigens during infection with C. burnetii, sera from mice experimentally infected with Nine Mile phase I were tested by immunoblotting. The mouse sera recognized antigens with a variety of molecular weights, including proteins of 14, 22, 28, 34, and 60 kDa as immunodominant antigens. In order to clone potential protective antigens, a genomic DNA library of Nine Mile phase I was constructed in the expression vector Lambda ZAP Express and screened with sera from mice that recovered from C. burnetii infection. A total of 102 immunoreactive clones with various signal intensities were identified from about 8,000 plaques. These clones were purified and expressed in the excised plasmid pBK-CMV. The proteins expressed by these recombinant plasmids were analyzed by SDS-PAGE and immunoblotting. Fifty-four clones expressed immunoreactive proteins of molecular masses ranging from approximately 14 to 60 kDa. Sequence analysis and BLAST search of the recently completed genome sequence identified a variety of novel immunoreactive proteins. These proteins are logical vaccine candidates for testing protective activity against C. burnetii challenge. We established a sublethal challenge model in BALB/c mice with protection from the development of severe splenomegaly as an indicator of vaccinogenic activity. Further characterization of these proteins will provide essential information for developing novel, specific diagnostic reagents and potential subunit vaccine candidates against C. burnetii infection.


Short-Chain Fatty Acids Protect Against High-Fat Diet-Induced Obesity via a PPARγ-Dependent Switch From Lipogenesis to Fat Oxidation.
Short-chain fatty acids (SCFAs) are the main products of dietary fiber fermentation and are believed to drive the fiber-related prevention of the metabolic syndrome. Here we show that dietary SCFAs induce a peroxisome proliferator-activated receptor-γ (PPARγ)-dependent switch from lipid synthesis to utilization. Dietary SCFA supplementation prevented and reversed high-fat diet-induced metabolic abnormalities in mice by decreasing PPARγ expression and activity. This increased the expression of mitochondrial uncoupling protein 2 and raised the AMP-to-ATP ratio, thereby stimulating oxidative metabolism in liver and adipose tissue via AMPK. The SCFA-induced reduction in body weight and stimulation of insulin sensitivity were absent in mice with adipose-specific disruption of PPARγ. Similarly, SCFA-induced reduction of hepatic steatosis was absent in mice lacking hepatic PPARγ. These results demonstrate that adipose and hepatic PPARγ are critical mediators of the beneficial effects of SCFAs on the metabolic syndrome, with clearly distinct and complementary roles. Our findings indicate that SCFAs may be used therapeutically as cheap and selective PPARγ modulators.


Comparison of the positive and negative ion electrospray ionization and matrix-assisted laser desorption ionization-time-of-flight mass spectra of the reaction products of phosphatidylethanolamines and hypochlorous acid.
Phosphatidylethanolamines (PEs) react with HOCl under formation of the mono- and dichloramines which are easily converted into secondary products (nitriles and imines). PEs with unsaturated acyl residues also give chlorhydrines. The aim of this study was to investigate whether all products may be detected by electrospray ionization (ESI) and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Results indicated that chloramines and imines are nearly exclusively detectable by ESI-MS, whereas all other products are detectable by both MALDI and ESI-MS. Therefore, ESI-MS is superior for the detection of chlorinated products of PEs.


Conjugated hyperbilirubinaemia in Nigerian infants.
A prospective study of 101 Nigerian infants with conjugated hyperbilirubinaemia seen over 6 years shows that extrahepatic biliary tract obstruction, idiopathic hepatitis, and bacterial infections were the common causes. A firm diagnosis was based on clinical, biochemical and histological features when the patient presented early. However, most of the infants presented late and the superimposed features of prolonged cholestasis made differentiation of the probable causes difficult. Erythrocyte peroxide haemolysis test and laparatomy aided diagnosis in these cases. Seventy five per cent of the patients with sepsis treated with antibiotics, and 70% of those with hepatitis treated symptomatically, recovered. Surgery was successful in only 15% of the patients with biliary tract obstruction. These were those who had either diverticulum of the common bile duct, localized atresia or stenosis or in whom biliary obstruction was due to viscid bile.


Persistant abnormal serum parathyroid hormone after renal transplantation: correlation between renal graft function.
Secondary hyperparathyroidism in uremia renal transplantation. We studied the i-PTH (immunoactive parathyroid hormone) of nineteen immunoactive parathyroid hormone) of nineteen patients before and after renal transplantation with average of 20 months post transplantation. In group I patients with normal i-PTH serum level had an significant lower serum level of creatinine than Group II with elevated abnormal i-PTH. While the calcium and phosphorus showed no significant difference between group I and Group II patients. We suggested that abnormal serum i-PTH level after renal transplantation after one year was closely related to renal graft function. This group of abnormal serum i-PTH should be closely monitored for their renal graft function.


Unidirectionally progressive left pneumonectomy & mediastinal lymph node dissection.
The patient has lower left lung tumor and adenocarcinoma at the openings of both upper and lower left lung. Preoperative bronchoscopic biopsy has confirmed the diagnosis. The surgical approach is unidirectionally progressive left pneumonectomy + mediastinal lymph node dissection. The layers of structure are treated one after another until the fissure from a single direction through the working port. Hence, the resecting order should be left superior pulmonary vein-left lower pulmonary vein-left main bronchus-left pulmonary artery. The vessels and bronchi are cut using an endoscopic linear stapler or the Hemolock clips. The resected lobe is placed into a large-size specimen bag and retrieved through the working port to prevent contamination of the chest incision by any tumor tissue. Mediastinal lymph node dissection is performed at the end. The surgery is performed under general anesthesia with double-lumen endotracheal intubation. The patient is placed in a 90-degree position lying on the unaffected side. Similar to traditional resection of left lung lobes, an approximately 1.5-cm observation port is created in the 7th intercostal space between the middle and anterior axillary lines, an approximately 4-cm working port in the 4th intercostal space between the anterior axillary line and the midclavicular line, and an approximately 1.5-cm auxiliary port in the 9th intercostal space between the posterior axillary line and the subscapular line. The operator stands in front of the patient, manipulating the endoscopic instruments while watching the monitor.


[Therapy and etiology of sebopsoriasis].
9 patients suffering from psoriasis vulgaris associated with sebopsoriasis lesions as well as 21 patients showing sebopsoriasis only were treated with 1% bifonazole cream and gel. We found good and excellent results in 86% of the sebopsoriatic, but only in 40% of the psoriasis vulgaris lesions. These results were achieved independently of the galenic preparation. In some of the sebopsoriasis patients, typical lesions were brought about by intracutaneous injection of bacterial antigens like steptococcus viridans and staphylococcus albus; my be, this is a hint on the etiology.


Mouse Paternal RNAs Initiate a Pattern of Metabolic Disorders in a Line-Dependent Manner.
A wide range of diseases result from environmental effects, and the levels of many native transcripts are altered. The alteration of non-coding RNAs (ncRNAs) and transmission of the variation to the next generation is increasingly recognized as a marker of disease. However, the determining signals and mechanisms of RNA-induced heritability remain unclear. We performed functional tests with four different genotypes of mice maintained on a high-fat diet to trace the transfer of the obesity/diabetes phenotype to the next generation in order to detect common signals. Two founders of four mouse lines (B6/D2 hybrid and Dnmt2 -/-C57BL/6 ) resist and do not change their phenotype while their sperm RNAs after microinjection into fertilized mouse eggs transfer the newly acquired phenotypes in a susceptible inbred line (C57BL/6 or Balb/c). Unexpectedly, in the same line of experiments, sperm RNA from animals raised on a normal diet when mixed with the sperm RNA from animals raised on a diet high in fat or synthetic miR-19b (inducer of obesity) affects or prevents the development of obesity and diabetes. However, it remains unclear what happens to ncRNA signaling under diet. With a comprehensive new analysis of the transcripts maintained as an RNA/DNA hybrid in sperm, we suggest that a fraction of the RNAs are stably attached to the genome. Thus, we propose that changes in the dynamics of ncRNA retention on DNA by factors such as transcriptional variations or lack of adequate methylation could serve as molecular markers to trace these epigenetics events.


Effects of high-fat diets with different carbohydrate-to-protein ratios on energy homeostasis in rats with impaired brain melanocortin receptor activity.
Changes in dietary macronutrient composition and/or central nervous system neuronal activity can underlie obesity and disturbed fuel homeostasis. We examined whether switching rats from a diet with high carbohydrate content (HC; i.e., regular chow) to diets with either high fat (HF) or high fat/high protein content at the expense of carbohydrates (LC-HF-HP) causes differential effects on body weight and glucose homeostasis that depend on the integrity of brain melanocortin (MC) signaling. In vehicle-treated rats, switching from HC to either HF or LC-HF-HP feeding caused similar reductions in food intake without alterations in body weight. A reduced caloric intake (-16% in HF and LC-HF-HP groups) required to maintain or increase body weight underlay these effects. Chronic third cerebroventricular infusion of the MC receptor antagonist SHU9119 (0.5 nmol/day) produced obesity and hyperphagia with an increased food efficiency again observed during HF (+19%) and LC-HF-HP (+33%) feeding. In this case, however, HF feeding exaggerated SHU9119-induced hyperphagia and weight gain relative to HC and LC-HF-HP feeding. Relative to vehicle-treated controls, SHU9119 treatment increased plasma insulin (2.8-4 fold), leptin (7.7-15 fold), and adiponectin levels (2.4-3.7 fold), but diet effects were only observed on plasma adiponectin (HC and LC-HF-HP<HF). Finally, SHU9119-treated LC-HF-HP-fed rats were less glucose tolerant than others. Relatively low plasma adiponectin levels likely contributed to this effect. Thus HF feeding amplifies obesity induced by impaired MC signaling, provided that the carbohydrate-to-protein (C/P) ratio is high enough. Reduction of the C/P ratio within a HF diet ameliorates hyperphagia and obesity in rats with impaired MC signaling but aggravates associated disturbances in fuel homeostasis.


Silymarin: not just another antioxidant.
Silymarin (Silybum marianum; SM), popularly known as milk thistle, is an extract that has been used for many centuries to treat liver diseases. In recent years, several studies have shown that SM is not only just another antioxidant but also a multifunctional compound that exhibits several beneficial properties for use in the treatment and prevention of different types of pathologies and disorders. This review aims at demonstrating the main protective activities of SM in diseases, such as cancer, diabetes, hepatitis, non-alcoholic fatty liver disease, alcoholic liver disease, hepatitis C virus, hepatitis B virus, metabolic syndrome, depression, cardiovascular diseases and thalassemia, in addition to its photoprotective activity in in vitro tests and preclinical studies. Its main functions include antioxidant and anti-inflammatory effects, and it acts as modulator of signaling pathways. It has been suggested that SM presents great multifunctional potential and is capable of achieving promising results in different types of research. However, caution is still needed regarding its indiscriminate use in humans as there are only a few clinical studies relating to the adequate dose and the actual efficacy of this extract in different types of diseases.


Identification of the Principle of Taste Sensors to Detect Non-Charged Bitter Substances by 1H-NMR Measurement.
A taste sensor with lipid/polymer membranes is attracting attention as a method to evaluate taste objectively. However, due to the characteristic of detecting taste by changes in membrane potential, taste sensors cannot measure non-charged bitter substances. Many foods and medicines contain non-charged bitter substances, and it is necessary to quantify these tastes with sensors. Therefore, we have been developing taste sensors to detect bitter tastes caused by non-charged substances such as caffeine. In previous studies, a sensor for detecting bitterness caused by caffeine and theobromine, theophylline, was developed, using a membrane modified with hydroxybenzoic acid (HBA) as the sensing part. The sensor was designed to form intramolecular hydrogen bonds (H-bonds) between the hydroxy group and carboxy group of HBA and to successively cause the intermolecular H-bonds between HBA and caffeine molecules to be measured. However, whether this sensing principle is correct or not cannot be confirmed from the results of taste sensor measurements. Therefore, in this study, we explored the interaction between HBA and caffeine by 1H-nuclear magnetic resonance spectroscopy (NMR). By the 1H NMR detection, we confirmed that both the substances interact with each other. Furthermore, the nuclear Overhauser effect (NOE) of intermolecular spatial conformation in solution was measured, by which 2,6-dihydroxybenzoic acid (2,6-DHBA) preferably interacted with caffeine via the H-bonding and stacking configuration between aromatic rings. Identifying the binding form of 2,6-DHBA to caffeine was estimated to predict how the two substances interact.


[Multiplicity of mitochondrial monoamine exidases and ways of selective blocking of the enzyme activity].
Problems on multiple forms of mitochondrial monoamine oxidases are considered. Possible nature of monoamine oxidase isoenzymes and its A and B forms is discussed. Substrates specificity of mitochondrial monoamine oxidase and the importance of amine chemical structure for the enzyme activity is examined. The data on the structure of mitochondrial monoamine oxidase active center and the presence in it of several regions responsible for amine binding are analyzed. The nature of these regions is discussed. New approaches to selective blocking of the activity of mitochondrial monoamine oxidase are considered on the basis of the hypothesis given on the mechanism of substrate and inhibitor interaction with the enzyme.


HIV counselling and testing utilisation and attitudes of male inmates in a South African prison.
The Department of Correctional Services Policy on the management of HIV and AIDS for offenders include voluntary counselling and testing (VCT) for HIV as one of the priorities in the rehabilitation of inmates. The aim of this study was to determine factors associated with the utilisation of VCT services in the correctional centres in terms of level of satisfaction, their experiences and expectations, and motivating factors and barriers for VCT utilisation at Losperfontein Correctional Centre, South Africa. This was a case control study (cases being those who underwent testing and controls those who did not) examining predictors of HIV VCT utilisation among 200 male adult sentenced inmates serving medium and maximum sentences. Results indicate that a poor health system (OR=0.34, 95%CI: 0.23 - 0.50) was inversely associated with HIV testing acceptance in prison, while age, educational level, population group, marital status, length of incarceration and access to HIV testing in prison were not associated with HIV testing acceptance in prison. Half of the participants (50%) agreed that VCT services are accessible and are promoted at their correctional centre. Most were satisfied with different components of VCT services, ranging from 79% (fair to very good) for 'the way he/she received you' to 62% 'clarified all your concerns'. This study demonstrated some challenges and benefits to the field of health promotion and HIV prevention in the correctional centres especially with regard to VCT services.


Impact of perioperative care on the post-operative recovery of women undergoing surgery for gynaecological tumours.
To assess perioperative care in patients undergoing abdominal surgery for gynaecological tumours and how it relates to post-operative (PO) complications and oral PO feeding. Ninety-one women undergoing major abdominal surgery for gynaecological tumours were enrolled. Data included mechanical bowel preparation (MBP), prescribed diet, length of fast, start date of oral diet and progression of food consistency, anaesthetic technique, use of opioids and intravenous hydration (IH). Outcomes evaluated were nausea, vomiting and abdominal distension. The median pre-operative length of fast was 11.4 h. PO digestive complications occurred in 46.2% of the patients. Median intraoperative total IH and crystalloids were significantly higher in patients with abdominal distension during the first and second PO day. MBP with mannitol implied greater intraoperative IH and was significantly associated with a higher incidence of immediate PO nausea. Post-operative IH was also associated with gastrointestinal complications. The best cut-off point for the cumulative fluid load PO for determining a longer PO hospital stay was 4 L. Performing MBP before surgery and excessive IH are factors related to major digestive complications in our study population. Changes in pre-operative fasting time and PO refeeding should be considered to reduce the gastrointestinal complications and PO recovery time.


Discovery of CC chemokine receptor-3 (CCR3) antagonists with picomolar potency.
Starting with our previously described(20) class of CC chemokine receptor-3 (CCR3) antagonist, we improved the potency by replacing the phenyl linker of 1 with a cyclohexyl linker and by replacing the 4-benzylpiperidine with a 3-benzylpiperidine. The resulting compound, 32, is a potent and selective antagonist of CCR3. SAR studies showed that the 3-acetylphenyl urea of 32 could be replaced with heterocyclic ureas or heterocyclic-substituted phenyl ureas and still maintain the potency (inhibition of eotaxin-induced chemotaxis) of this class of compounds in the low-picomolar range (IC(50) = 10-60 pM), representing some of the most potent CCR3 antagonists reported to date. The potency of 32 for mouse CCR3 (chemotaxis IC(50) = 41 nM) and its oral bioavailability in mice (20% F ) were adequate to assess the efficacy in animal models of allergic airway inflammation. Oral administration of 32 reduced eosinophil recruitment into the lungs in a dose-dependent manner in these animal models. On the basis of its overall potency, selectivity, efficacy, and safety profile, the benzenesulfonate salt of 32, designated DPC168, entered phase I clinical trials.


Proteolytic stress causes heat shock protein induction, tau ubiquitination, and the recruitment of ubiquitin to tau-positive aggregates in oligodendrocytes in culture.
Molecular chaperones and the ubiquitin-proteasome system are participants in the defense against unfolded proteins and provide an effective protein quality control system that is essential for cellular functions and survival. Ubiquitinated tau-positive inclusion bodies containing the small heat shock protein alphaB-crystallin in oligodendrocytes are consistent features of a variety of neurodegenerative diseases, and defects in the proteasome system might contribute to the aggregation process. Oligodendrocytes, the myelin-forming cells of the CNS, are specifically sensitive to stress situations. Here we can show that in cultured rat brain oligodendrocytes proteasomal inhibition by MG-132 or lactacystin caused apoptotic cell death and the induction of heat shock proteins in a time- and concentration-dependent manner. Specifically, alphaB-crystallin was upregulated, and ubiquitinated proteins accumulated. After incubation with MG-132 the tau was dephosphorylated, which enhanced its microtubule-binding capacity. Proteasomal inhibition led to ubiquitination of tau and its association with alphaB-crystallin and to the occurrence of thioflavine S-positive aggregates in the oligodendroglial cytoplasm. These aggregates were positive for tau and also contained ubiquitin and alphaB-crystallin; hence they resembled the glial cytoplasmic inclusions observed in white matter disease and frontotemporal dementias with parkinsonism linked to chromosome 17 (FTDP-17). In summary, the data underscore the specific sensitivity of oligodendrocytes to stress situations and point to a causal relationship of proteasomal impairment and inclusion body formation.


Trends in Infective Endocarditis Mortality in the United States: 1999 to 2020: A Cause for Alarm.
Data on national trends in mortality due to infective endocarditis (IE) in the United States are limited. Utilizing the multiple causes of death data from the Centers for Disease Control and Prevention's Wide-Ranging Online Data for Epidemiologic Research database from 1999 to 2020, IE and substance use were identified using the International Classification of Diseases, Tenth Revision, Clinical Modification codes. Between 1999 and 2020, the IE-related age-adjusted mortality rates declined. IE-related crude mortality accelerated significantly in the age groups 25-34 years (average annual percentage change, 5.4 [95% CI, 3.1-7.7]; P<0.001) and 35-44 years (average annual percentage change, 2.3 [95% CI, 1.3-3.3]; P<0.001), but remained stagnant in those aged 45-54 years (average annual percentage change, 0.5 [95% CI, -1.9 to 3]; P=0.684), and showed a significant decline in those aged ≥55 years. A concomitant substance use disorder as multiple causes of death in those with IE increased drastically in the 25-44 years age group (P<0.001). The states of Kentucky, Tennessee, and West Virginia showed an acceleration in age-adjusted mortality rates in contrast to other states, where there was predominantly a decline or static trend for IE. Age-adjusted mortality rates due to IE in the overall population have declined. The marked acceleration in mortality in the 25- to 44-year age group is a cause for alarm. Regional differences with acceleration in IE mortality rates were noted in Kentucky, Tennessee, and West Virginia. We speculate that this acceleration was likely due mainly to the opioid crisis that has engulfed several states and involved principally younger adults.


Angioside: The role of Angiogenesis and Hypoxia in Lung Neuroendocrine Tumours According to Primary Tumour Location in Left or Right Parenchyma.
Well-differentiated lung neuroendocrine tumours (Lu-NETs), classified as typical (TC) and atypical (AC) carcinoids, represent 30% of NETs. Angiogenesis plays an essential role in NET development and progression. A higher vascular network is a marker of differentiation, with positive prognostic implications. Materials and Methods: We retrospectively evaluated microvessel density (MVD) by CD34 immunohistochemical (IHC) staining and hypoxia by IHC staining for Hypoxia-inducible factor 1α (HIF-1α), comparing right- and left-lung parenchyma in 53 lung NETs. Results: The median age was 66 years (39−81), 56.6% males, 24.5% AC, 40.5% left-sided tumours and 69.8% TNM stage I. The mitotic count was <2/10 per 10 HPF in 79.2%, and the absence of necrosis in 81.1%, 39.6% with Ki67, was ≤2%. The MVD, the number of vessels and the average vessel area median values were significantly higher in the right than the left parenchyma (p: 0.025, p: 0.019, p: 0.016, respectively). Hypoxia resulted present in 14/19 (73.6%) left tumours and in 10/20 (50%) right tumours in the parenchyma (p: 0.129). Conclusions: This study suggests a biological rationale for a different angiogenesis and hypoxia according to the Lu-NETs' location. In our study, left primary tumours were less vascularized and most likely to present hypoxia than right primary tumours. This finding could have potentially useful prognostic and predictive implications for Lu-NETs.


Signaling Cascade through DC-ASGPR Induces Transcriptionally Active CREB for IL-10 Induction and Immune Regulation.
The types and magnitude of Ag-specific immune responses can be determined by the functional plasticity of dendritic cells (DCs). However, how DCs display functional plasticity and control host immune responses have not been fully understood. In this study, we report that ligation of DC-asialoglycoprotein receptor (DC-ASGPR), a C-type lectin receptor (CLR) expressed on human DCs, resulted in rapid activation of Syk, followed by PLCγ2 and PKCδ engagements. However, different from other Syk-coupled CLRs, including Dectin-1, signaling cascade through DC-ASGPR did not trigger NF-κB activation. Instead, it selectively activated MAPK ERK1/2 and JNK. Rapid and prolonged phosphorylation of ERK1/2 led to sequential activation of p90RSK and CREB, which consequently bound to IL10 promoter and initiated cytokine expression. In addition, DC-ASGPR ligation activated Akt, which differentially regulated the activities of GSK-3α/β and β-catenin and further contributed to IL-10 expression. Our observations demonstrate that DC-ASGPR induces IL-10 expression via an intrinsic signaling pathway, which provides a molecular explanation for DC-ASGPR-mediated programing of DCs to control host immune responses.


Multicomponent intervention to improve blood pressure management in chronic kidney disease: a protocol for a pragmatic clinical trial.
The purpose of this study is to incorporate behavioural economic principles and user-centred design principles into a multicomponent intervention for the management of uncontrolled hypertension (HTN) in chronic kidney disease (CKD) in primary care. This is a multicentre, pragmatic, controlled trial cluster-randomised at the clinician level at The Brigham and Women's Practice -Based Research Network of 15 practices. Of 220 total clinicians, 184 were eligible to be enrolled, and the remainder were excluded (residents and clinicians who see urgent care or walk-in patients); no clinicians opted out. The intervention consists of a clinical decision support system based in behavioural economic and user-centred design principles that will: (1) synthesise existing laboratory tests, medication orders and vital sign data; (2) increase recognition of CKD, (3) increase recognition of uncontrolled HTN in CKD patients and (4) deliver evidence-based CKD and HTN management recommendations. The primary endpoint is the change in mean systolic blood pressure between baseline and 6 months compared across arms. We will use the Reach Effectiveness Adoption Implementation Maintenance framework. At the conclusion of this study, we will have: (1) validated an intervention that combines laboratory tests, medication records and clinical information collected by electronic health records to recognise uncontrolled HTN in CKD patients and recommend a course of care, (2) tested the effectiveness of said intervention and (3) collected information about the implementation of the intervention that will aid in dissemination of the intervention to other practice settings. The Human Subjects Institutional Review Board at Brigham and Women's Hospital provided an expedited review and approval for this study protocol, and a Data Safety Monitoring Board will ensure the ongoing safety of the trial. NCT03679247.


[Electron-microscopic and autoradiographic study of giant cells of foreign bodies in the focus of aseptic inflammation].
Ultrastructure of sinuous proximal and straight distal tubules, as well as collecting tubules of the cortical layer in the rat kidney fixed with perfusion has been studied with electron microscopic morphometry 7--8 weeks after contralateral nephrectomy. The volume of mitochondria, the area of their crists, the area of the membranes in intracellular labyrinth and other morphometrical parameters have been calculated. Relative volume of mitochondria in the nephron areas studied does not change, while in the collecting tubules it is elevated. The area of crists in mitochondria and "coefficient of morphological organization level" of these organells in the proximal tubules are increased, in the distal do not change, in the collecting tubules increase again. Subcellular changes described are discussed mainly in terms of enhancement of concentrating function of the compensatory-hypertrophic kidney.


Development of a multiplex real-time PCR assay using two thermocycling platforms for detection of major bacterial pathogens associated with bovine respiratory disease complex from clinical samples.
Bovine respiratory disease complex (BRDC) is one of the most significant diseases of cattle. Bacterial pathogens involved in BRDC include Mannheimia haemolytica, Mycoplasma bovis, Histophilus somni, and Pasteurella multocida. We developed and evaluated a multiplexed real-time hydrolysis probe (rtPCR) assay using block-based Peltier and rotary-based thermocycling on lung tissue, nasal swabs, and deep nasopharyngeal swabs. The rtPCR results were compared to culture or a gel-based M. bovis PCR using statistical analysis to determine optimum quantification cycle (Cq) cutoffs to maximize agreement. The limits of detection were 1.2-12 CFU/reaction for each pathogen. M. haemolytica was the most prevalent organism detected by rtPCR, and was most frequently found with P. multocida. The rtPCR assay enabled enhanced levels of detection over culture for all pathogens on both thermocycling platforms. The rotary-based thermocycler had significantly lower Cq cutoffs (35.2 vs. 39.7), which maximized agreement with gold standard culture or gel-based PCR results following receiver operating characteristic analysis to maximize sensitivity (Se) and specificity (Sp). However, overall assay Se and Sp were similar on both platforms (80.5% Se, 88.8% Sp vs. 80.1% Se, 88.3% Sp). Implementation of these tests could enhance the detection of these pathogens, and with high-throughput workflows could reduce assay time and provide more rapid results. The assays may be especially valuable in identifying coinfections, given that many more antemortem samples tested in our study were positive for 2 or more pathogens by rtPCR ( n = 125) than were detected using culture alone ( n = 25).


Lower all-cause, cardiovascular, and cancer mortality in centenarians' offspring.
To assess the cause of death for centenarians' offspring and controls. Cross-sectional study. Community-based, nationwide sample. Family pedigree information was collected on 295 offspring of centenarians (from 106 families with a parent already enrolled in the nationwide New England Centenarian Study) and on 276 controls (from 82 control families) from 1997 to 2000. Controls were individuals whose parents were born in the same year as the centenarians but at least one of whom died at the average life expectancy. Age at death and cause of death. Centenarians' offspring had a 62% lower risk of all-cause mortality (P<.001), a 71% lower risk of cancer-specific mortality (P=.002), and an 85% lower risk of coronary heart disease-specific mortality (P<.001). Significant differences were not found for other causes of death. However of those who died centenarian offsprings dead at a significantly younger age than controls. These findings suggest that centenarians' offspring have lower all-cause mortality rates and cause-specific mortality rates for cancer and coronary heart disease. These results suggest that mechanisms for survival to exceptional old age may go beyond the avoidance or delay of cardiovascular disease and also include the avoidance or delay of cancer. Moreover survival advantage of centenarian offsprings may not be due to factors related to childhood mortality. Ultimately, survival to exceptional old age may involve lower susceptibility to a broad range of age-related diseases, perhaps secondary to inhibition of basic mechanisms of aging.


Development and psychometric testing of the safety climate instrument suitable for nurses handling chemotherapy drugs.
To develop and test the psychometric properties of the Taiwanese safety climate instrument suitable for nurses handling chemotherapy drugs. This is an instrument development study. All four stages, including questionnaire design, expert consultation, cognitive testing and psychometric validation, were used in this study. The data were collected between August and December 2018. Nurses with experience in handling chemotherapy drugs (N = 484) at one medical centre and two regional hospitals in Taiwan completed this instrument. Data were randomly split into two groups: one group (N = 237) for exploratory factor analysis and the other (N = 247) for confirmatory factor analysis. The instruments' items were based on qualitative research, and the content validity index levels exceeded the acceptable value. An exploratory factor analysis revealed 43 items remaining in six factors, which accounted for 74.4% of variance. The result of the confirmatory factor analysis verified the acceptability of a 43-item model. The composite reliability values, Cronbach's alpha values, convergent validity and discriminant validity for each factor exceeded the acceptable value. Most climate safety instruments used in the health care sector focus mainly on patient safety outcomes. Furthermore, there is no safety climate instrument for handling chemotherapy drugs, and there is a cultural difference. Through the development and validation process, we have developed a new instrument suitable for nurses handling chemotherapy drugs, which has good psychometric properties. This instrument is valuable as its development was based on the concept of a safety climate for health care perceptions and qualitative survey findings. Hospital managers can use this instrument regularly to evaluate nurses' perceptions of the safety climate to determine the strengths and weaknesses of their workplace, thereby assisting organizational managers in proposing concrete actions.


Classification of Pseudomonas aeruginosa O antigens by immunoelectrophoresis.
Heated saline extracts of 89 strains, and (1) supernates of phenol-water extracts (L1 fractions), (2) purified lipopolysaccharide, (3) trichloracetic-acid (TCA) extracts, and (4) sodium-hydroxide extracts of 23 strains representing all Pseudomonas aeruginosa O antigens were subjected electrophoresis. Precipitation lines obtained with homologous and heterologous antisera were evaluated by electrodensitometric measurement. The characteristics of the immunoelectrophoretic groups established were as follows. Group I: two lines running at different rates towards the anode; three subgroups on the basis of the behaviour of alkali-treated antigens. Group II: triple line at the starting well, alkali sensitive. Group III: triple line at the starting well, alkali resistant; two subgroups according to reactivity or non-reactivity of L1 fractions. Group IV: triple line on the cathode side, alkali resistant, L1 fraction non-reactive. Group V: single line on the anode side, alkali sensitive, L1 fraction and TCA extract non-reactive. O antigens identified by agglutination corresponded closely with the immunoelectrophoretic pattern: strains with identical O antigens or sharing major somatic components fell, with one exception, into the same immunoelectrophoretic group.


Medication-Related Osteonecrosis of the Jaw: A Systematic Review and a Bioinformatic Analysis.
The objective was to evaluate the current evidence regarding the etiology of medication-related osteonecrosis of the jaw (MRONJ). This study systematically reviewed the literature by searching PubMed, Web of Science, and ProQuest databases for genes, proteins, and microRNAs associated with MRONJ from the earliest records through April 2023. Conference abstracts, letters, review articles, non-human studies, and non-English publications were excluded. Twelve studies meeting the inclusion criteria involving exposure of human oral mucosa, blood, serum, saliva, or adjacent bone or periodontium to anti-resorptive or anti-angiogenic agents were analyzed. The Cochrane Collaboration risk assessment tool was used to assess the quality of the studies. A total of 824 differentially expressed genes/proteins (DEGs) and 22 microRNAs were extracted for further bioinformatic analysis using Cytoscape, STRING, BiNGO, cytoHubba, MCODE, and ReactomeFI software packages and web-based platforms: DIANA mirPath, OmicsNet, and miRNet tools. The analysis yielded an interactome consisting of 17 hub genes and hsa-mir-16-1, hsa-mir-21, hsa-mir-23a, hsa-mir-145, hsa-mir-186, hsa-mir-221, and hsa-mir-424. A dominance of cytokine pathways was observed in both the cluster of hub DEGs and the interactome of hub genes with dysregulated miRNAs. In conclusion, a panel of genes, miRNAs, and related pathways were found, which is a step toward understanding the complexity of the disease.


Bright hot impacts by erupted fragments falling back on the Sun: a template for stellar accretion.
Impacts of falling fragments observed after the eruption of a filament in a solar flare on 7 June 2011 are similar to those inferred for accretion flows on young stellar objects. As imaged in the ultraviolet (UV)-extreme UV range by the Atmospheric Imaging Assembly onboard the Solar Dynamics Observatory, many impacts of dark, dense matter display uncommonly intense, compact brightenings. High-resolution hydrodynamic simulations show that such bright spots, with plasma temperatures increasing from ~10(4) to ~10(6) kelvin, occur when high-density plasma (>>10(10) particles per cubic centimeter) hits the solar surface at several hundred kilometers per second, producing high-energy emission as in stellar accretion. The high-energy emission comes from the original fragment material and is heavily absorbed by optically thick plasma, possibly explaining the lower mass accretion rates inferred from x-rays relative to UV-optical-near infrared observations of young stars.


[Rheumatism and urogenital infection].
The antiinfectious treatment of rheumatic diseases is certainly very old, but it has gained increased significance as a result of new bacteriological and immunological findings. Urogenital infection as a possible trigger mechanism in reactive arthritides and ankylosing spondylitis was investigated particularly thoroughly. Many experimental, immunological and clinical results indicate a pathogenetic role of infections with chlamydia or mycoplasma in inflammatory rheumatic diseases. However, numerous tests on cultures proved negative, and many serological-immunological findings are inconsistent. The "link" between urogenital infection and rheumatic diseases is still unknown. Of particular interest are the relations between chlamydial prostatitis and ankylosing spondylitis. Earlier findings on the induction of osteogenesis from undifferentiated connective tissue cells by epithelial cells from the prostate could be a basis for further research.


Mutation of Pseudomonas aeruginosa to piperacillin resistance mediated by beta-lactamase production.
Piperacillin-resistant mutants were detected in each of ten strains of Pseudomonas aeruginosa. The resistance was associated with constitutive production by the strains of high levels of beta-lactamase which was similar in character to the induced chromosomal Id beta-lactamase of the parent strains. This type of mutation to piperacillin-resistance occurred at a low frequency. The beta-lactamase production and piperacillin-resistance were stable on repeated subculture and prolonged storage of the strains.


Human rotavirus type 2: cultivation in vitro.
A strain of type 2 human rotavirus (Wa) was grown to relatively high titer through 14 passages in primary cultures of African green monkey kidney (AGMK) cells. This passage series was initiated with virus that had been passaged 11 times serially in newborn gnotobiotic piglets. In contrast, virus present in the stool of patient Wa as well as virus from the first, second, or third passage in piglets could not be propagated successfully in African green monkey kidney cells. Prior to each passage in cell culture, the virus was treated with trypsin and the inoculated cultures were centrifuged at low speed. Cultivation of a type 2 human rotavirus should aid attempts to characterize this virus and to develop a means of immunoprophylaxis for a serious diarrheal disease of human infants.


Noise analysis in air-coupled PVDF ultrasonic sensors.
In this paper we analyze the noise generated in a piezo-polymer based sensor for low frequency ultrasound in air. The sensor includes two curved PVDF transducers for medium and short range applications. A lumped RLC equivalent circuit was derived from the measurement of the transducer's electrical admittance, in air, by taking into account both mechanical and dielectric losses, which we suppose are the major sources of noise in similar devices. The electrical model was used to study and optimize the noise performance of a 61 kHz transducer and to simulate the electrical behavior of the complete transmitter-receiver system. The validity of the overall electrical model with low noise was confirmed after verifying, with Pspice, agreement of the practical and theoretical results.


Cognitive status, fast walking speed and walking speed reserve-the Gait and Alzheimer Interactions Tracking (GAIT) study.
The aims of this study were to (1) determine if older people at their fast walking speed (FWS) are able to reach the speed required at pedestrian crossings (>1.2 m/s) and (2) determine the role of cognitive impairment on the ability to alter speed and walk quickly. Participants were recruited from the Angers Memory Clinic, France. Gait speed was assessed at preferred and FWS using a GAITRite walkway. Walking speed reserve (WSR) was calculated as the difference between FWS and preferred speeds. Participants were classified into cognitive stages (cognitively healthy, mild cognitive impairment, mild and moderate dementia) based on neuropsychological evaluations. The proportion of participants with a FWS of <1.2 m/s was reported. The association between cognitive stage and preferred, fast and walking speed reserve was assessed using multivariable regression, adjusting for covariates. The mean age of the sample (n = 681) was 73.3 (SD 5.8) years. At preferred speed 73.7%, and at FWS 12.8%, of participants had speeds less than 1.2 m/s. Poorer cognitive stage was associated with slower preferred speed (β -0.08, 95% CI -0.10, -0.06), FWS (β -0.13, 95% CI -0.16, -0.10) and also with smaller WSR (m/s) (β -0.05, 95% CI -0.07, -0.03), but not WSR (%) (β -1.73, 95% CI -4.38, 0.93). In older people, worse stages of cognitive impairment were associated with poorer ability to increase speed and walk quickly. Such limitations may result in reduced ability to access the community.


Chronic nicotine pretreatment protects the blood-brain barrier against nicotine-induced seizures in the rat.
This study was designed to investigate the possible protective actions of nicotine on cerebrovascular permeability in convulsions during nicotine-induced seizures. We have measured the permeability changes in the blood-brain barrier (BBB) macroscopically and spectrophotometrically by using Evans blue dye. Specific gravity measurements were also performed to assess brain edema which develops after blood-brain barrier opening. The experiments were carried out on Wistar rats. Rats were divided into two groups. They received acutely a convulsive dose of nicotine 3, 5, 8 and 9 mg kg(-1)i.p. or pretreated with a low dose of nicotine (0.8 mg kg(-1)i. p.) for 21 days followed by the procedure mentioned in the first group. Acute nicotine injection induced a significant increase in blood pressure and Evans-blue passage, despite a decline in specific gravity values. Low doses of chronic nicotine administration markedly reduced both the leakage of dye, and brain water content. Chronic treatment with low doses of nicotine (0.8 mg kg(-1)day(-1)s. c.) lessened the intensity of tonic-clonic seizures observed with a single dose of 3, 5, 8 or 9 mg kg(-1)nicotine. The data presented here demonstrate that nicotine pretreatment results in decreased sensitivity to nicotine-induced seizures in rats.


Molecular characterization of the Tobacco rattle virus RNA2 genome isolated from Gladiolus.
Tobacco rattle virus (TRV-K) was first identified in a symptomatic Gladiolus plant cultivated in Korea. We analyzed the TRV-K genome and compared its phylogeny with other TRV isolates. After constructing of a full-length genomic RNA2 strand clone, a complete sequence was generated from several overlapping clones. The cloned genome was 3261 bases in length, identical to TRV-K, and had three open reading frames. TRV-K had the highest sequence identity with the American isolate TRV-ORY. Sequence analysis of the RNA2 genome showed that TRV-K contains an intact 2a, 2b, and 2c coding sequence and an RNA1-related 3' terminus, which is typical of TRV RNA2. Phylogenetic analysis revealed that TRV-K is in the same cluster as the American isolates and another Korean isolate, TRV-SK; however, it was in a different cluster than the European isolates.


Adult onset tic disorders.
Tic disorders presenting during adulthood have infrequently been described in the medical literature. Most reports depict adult onset secondary tic disorders caused by trauma, encephalitis, and other acquired conditions. Only rare reports describe idiopathic adult onset tic disorders, and most of these cases represent recurrent childhood tic disorders. To describe a large series of patients with tic disorders presenting during adulthood, to compare clinical characteristics between groups of patients, and to call attention to this potentially disabling and underrecognised neurological disorder. Using a computerised database, all patients with tic disorders who presented between 1988 and 1998 to the movement disorders clinic at Columbia-Presbyterian Medical Center after the age of 21 were identified. Patients' charts were retrospectively reviewed for demographic information, age of onset of tics, tic phenomenology, distribution, the presence of premonitory sensory symptoms and tic suppressibility, family history, and associated psychiatric features. These patients' videotapes were reviewed for diagnostic confirmation and information was obtained about disability, course, and response to treatment in a structured follow up interview. Of 411 patients with tic disorders in the database, 22 patients presented for the first time with tic disorders after the age of 21. In nine patients, detailed questioning disclosed a history of previous childhood transient tic disorder, but in 13 patients, the adult onset tic disorder was new. Among the new onset cases, six patients developed tics in relation to an external trigger, and could be considered to have secondary tic disorders. The remaining patients had idiopathic tic disorders. Comparing adult patients with recurrent childhood tics and those with new onset adult tics, the appearance of the tic disorder, the course and prognosis, the family history of tic disorder, and the prevalence of obsessive-compulsive disorder were found to be similar. Adults with new onset tics were more likely to have a symptomatic or secondary tic disorder, which in this series was caused by infection, trauma, cocaine use, and neuroleptic exposure. Adult onset tic disorders represent an underrecognised condition that is more common than generally appreciated or reported. The clinical characteristics of adults newly presenting to a movement disorder clinic with tic disorders are reviewed, analysed, and discussed in detail. Clinical evidence supports the concept that tic disorders in adults are part of a range that includes childhood onset tic disorders and Tourette's syndrome.


Anatomy of the mandibular canal and surrounding structures. Part II: Cancellous pattern of the mandible.
It is necessary to correlate cancellous bone patterns with cone beam computed tomography (CBCT) images, but this has not been done to date. The goal of this study was to establish how the superior wall of the mandibular canal (MC) on CBCT images correlates with the cancellous bone around the MC on gross anatomical findings. Twenty sides from 10 dry mandibles derived from six females and four males were used for this study. In order to observe the MC distally, the specimen was prepared by the method used in our previous study. The cancellous bone around the MC was observed and classified into three types: type I (trabecular pattern), type II (osteoporotic pattern), and type III (dense/irregular pattern). The mandibles were examined with CBCT and the superior wall of the MC on CBCT was scored as visible or non-visible. Finally, the scores (visible or non-visible) were compared to the type by gross observation. For gross observation, a total of 80 areas were available for this study. The data were added to those from our previous study. As a result, 155 areas on 40 sides were analyzed. In dentulous sections, types I, II, and III were found in 55.8%, 20.9%, and 23.3%, respectively. In edentulous sections, the corresponding percentages were 25.0%, 41.1%, and 33.9%, respectively. The dentulous sections was more likely than edentulous to have a type I mandible in both sexes. More females than males had type II (osteoporotic) mandibles. When the superior wall of the MC was non-visible on CBCT, the cancellous bone was type II in 80%. We believe the results can easily be applied to preoperative diagnosis with not only radiological but also anatomical evidence. This classification now necessitates clinical trials for further evidence.


Data Collection from Buried Sensor Nodes by Means of an Unmanned Aerial Vehicle.
The development of Wireless Underground Sensor Networks (WUSNs) is a recent research axis based on sensor nodes buried a few dozen centimeters deep. The communication ranges are, however, highly reduced due to the high attenuation of electromagnetic waves in soil, leading to issues of data collection. This paper proposes to embed a data collector on an Unmanned Aerial Vehicle (UAV) coming close to each buried sensor node. The whole system was developed (sensor nodes, data collector, gateway) and experimentations were carried out in real conditions. In hovering mode, the measurements on the RSSI levels with respect to the position of the UAV highlight the interest in maintaining a high altitude when the UAV is far from the node. In dynamic mode, the experimental results demonstrate the feasibility of carrying out the data collection task while the UAV is moving. The speed of the UAV has, however, to be adapted to the required time to collect the data. In the case of numerous buried sensor nodes, evolutionary algorithms are implemented to plan the trajectory of the UAV optimally. To the best of our knowledge, this paper is the first one that reports experiment results combining WUSN and UAV technologies.


Lupus carditis: evaluation with technetium-99m MIBI myocardial SPECT and heart rate variability.
The objective of this paper was to investigate the incidence of myocardial perfusion defects in patients with systemic lupus erythematosus (SLE) associated with dysautonomic alterations. Twenty patients without any sign or symptoms of heart disease, selected from a larger population of patients with SLE, underwent technetium-99m sestamibi (Tc-99m MIBI) single photon emission computed tomography (SPECT), at rest and after dipyridamole infusion; they also underwent heart rate variability (HRV) examination by a 24 hour ambulatory electrocardiography, analyzing in the time domain the standard deviation of the R-R intervals average (SDNN) and the percentage of R-R adjacent intervals differing from each other more than 50 msec (pNN50); in the frequency domain the low (LF) and high frequencies (HF) were analyzed. Twenty healthy volunteers served as control group for heart rate variability. At MIBI-SPECT examination, the scan was found abnormal in 15 patients and normal in five: three patients demonstrated reversible defects in the anteroseptal region, four had irreversible defects in a region (two in the anteroseptal region and two in the lateral region), two had rest defects in two different regions (lateral and inferior, lateral and anteroseptal) that improved during dipyridamole scan, and six had both reversible and irreversible defects: four in a single segment (three anteroseptal and one lateral, and two in two different regions, particularly anteroseptal and lateral, lateral and inferior). All 20 patients showed significantly lower HRV parameters in comparison with the control group, except for pNN50, which indicates decreased physiologic periodic fluctuations of the autonomic nervous system. In six patients who underwent coronary angiography, the epicardial vessels were found completely normal. In view of the high incidence of myocardial hypoperfusion in patients with HRV alterations, the authors hypothesize that autonomic dysfunction may be associated with microvascular disease or metabolic alteration. They also believe that MIBI scintigraphy is a suitable technique in detecting myocardial damage in SLE patients free of clinical manifestation.


Multi-view stereo analysis reveals anisotropy of prestrain, deformation, and growth in living skin.
Skin expansion delivers newly grown skin that maintains histological and mechanical features of the original tissue. Although it is the gold standard for cutaneous defect correction today, the underlying mechanisms remain poorly understood. Here we present a novel technique to quantify anisotropic prestrain, deformation, and growth in a porcine skin expansion model. Building on our recently proposed method, we combine two novel technologies, multi-view stereo and isogeometric analysis, to characterize skin kinematics: Upon explantation, a unit square retracts ex vivo to a square of average dimensions of [Formula: see text]. Upon expansion, the unit square deforms in vivo into a rectangle of average dimensions of [Formula: see text]. Deformations are larger parallel than perpendicular to the dorsal midline suggesting that skin responds anisotropically with smaller deformations along the skin tension lines. Upon expansion, the patch grows in vivo by [Formula: see text] with respect to the explanted, unexpanded state. Growth is larger parallel than perpendicular to the midline, suggesting that elevated stretch activates mechanotransduction pathways to stimulate tissue growth. The proposed method provides a powerful tool to characterize the kinematics of living skin. Our results shed light on the mechanobiology of skin and help us to better understand and optimize clinically relevant procedures in plastic and reconstructive surgery.


A meta-analysis of implant-based breast reconstruction and timing of adjuvant radiation therapy.
Implant-based breast reconstruction is the most common type of reconstruction after postmastectomy radiation therapy (PMRT). The impact of the timing of PMRT to a tissue expander or permanent implant is not well understood. The purpose of this systematic review was to evaluate outcomes in implant-based reconstruction and the timing of PMRT. A review of the English literature in the PubMed/MEDLINE database (2000-2016) was performed to identify all articles on implant-based breast reconstruction and PMRT. Cases from each study were grouped by PMRT to a tissue expander or PMRT to a permanent implant. Outcomes of interest included reconstructive failure and capsular contracture as overall rates and associations were pooled. Effect sizes (z values), risk ratios (RRs), and heterogeneity scores (I2) were calculated on meta-analysis. There were 20 studies meeting inclusion criteria with 2348 patients identified. Pooled analysis revealed an overall rate of reconstructive failure of 17.6% and Baker grade III/IV capsular contracture of 37.5%. PMRT applied to tissue expanders resulted in higher rates of reconstructive failure compared with PMRT applied to permanent silicone implants (20% versus 13.4%, RR = 2.33, P = 0.0083, 95% confidence interval 1.24-4.35), but lower rates of capsular contracture (24.5% versus 49.4%, RR = 0.53, P = 0.083, 95% confidence interval 0.26-1.09). Regardless of timing, PMRT applied to implant-based breast reconstruction was associated with high risk of reconstructive failure and capsular contracture. Surgeons should consider alternative strategies, such as autologous tissue reconstructions, in patients requiring PMRT.


The Effects of Musculoskeletal Disorders on Professional Quality of Life Among Midwives Working in Delivery Rooms.
This study was conducted to determine the effects of musculoskeletal disorders on professional quality of life among midwives working in delivery rooms. The aim of the study was to determine the effect of musculoskeletal disorders of midwives working in delivery rooms on the quality of professional life. This descriptive study was conducted with 88 midwives who worked in delivery rooms and delivered babies. The data were collected using the Nordic Musculoskeletal Questionnaire (NMQ) and the Professional Quality of Life Scale (ProQOL R-IV). The data were analysed with IBM SPSS V23 using MANOVA and path analysis. Of the midwives, 80.7% reported that they had pain in the waist in the last year. According to the result of path analysis for NMQ and ProQOL R-IV, only the path coefficient between the 12-month disability and burnout was found to be statistically significant (p = 0.012). The multivariate analysis showed that single midwives had higher job satisfaction than married ones (p = 0.030) and those who chose their profession willingly had higher job satisfaction than others (p = 0.043). It was observed that most of the midwives suffered from low back pain. Additionally, there may be a relationship between musculoskeletal problems experienced by midwives and their professional burnout levels. Relevant strategies to reduce occupational risks, especially musculoskeletal disorders, and to increase the professional quality of life of midwives should be implemented urgently.


The potency of siRNA-mediated growth inhibition following silencing of essential genes is dependent on siRNA design and varies with target sequence.
Silencing genes essential for replication and division using siRNA has potential as a therapeutic strategy for cancer treatment. In order to identify the most potent siRNA, target sequence and siRNA design must be considered together, as tolerance for structural changes can be sequence-dependent. Here we have used growth inhibition assays to investigate the effects of silencing of RRM1, RRM2, and PLK1 with standard siRNAs, Stealth() duplexes, and Dicer substrate siRNAs. The growth inhibitory effect of RRM1, RRM2, or PLK1 knockdown in A549 cells varied with mRNA target site and the format of the siRNA, with longer modified siRNAs generally more effective than standard siRNAs specific for the same target site. Standard siRNAs of varying activity became more potent inhibitors of growth when converted to Stealth() duplexes, and the increase in activity was due to a combination of chemical modification and length. In each case, the effect on activity of changing the siRNA format depended on the siRNA sequence. Taken together these results suggest that, in vitro, longer siRNAs with chemical modifications are in general more active than standard siRNAs targeting the same site, and that structure, chemical modification, and target site must be considered together to identify the most active siRNAs.


Recent patents on genes and gene sequences useful for developing breast cancer detection systems.
Breast cancer is the most prevalent disease and second leading cause of death among women in many countries. Mutations and consequent DNA damage in several genes such as oncogenes and tumour suppressor genes have been implicated with abnormal behavior of somatic cells, resulting into malignant growth termed cancer. DNA damage/changes in chromosomal DNA and failure of DNA repair mechanism are important factors for cancer causation. Aberrant behavior of cells leading to uncontrolled proliferation causing cancer is attributable to amplicon formation, large deletions in some genes, mutations, and recombination breakpoints etc. which cause aberrant gene expression. Several patented genes found to be associated with breast cancer, have been discussed, such as BRCA1, BRCA2, DAP kinase, MYH, BCSGs, BCW 2 and Id-2 etc. Several of the genes/markers associated with breast cancer can be considered as appropriate candidates for developing early detection systems/protocols for breast cancer.


[Problem of transient dementias of old age (clinico-diagnostic aspect)].
On the basis of the literature data and their own findings the authors discuss the problem of "transient" senile dementia. A special emphasis is given to depressive pseudodementias and somatogenic confusion imitating oligophrenia, particularly to their differentiation from true oligophrenia. The authors also describe supplementary differential-diagnostic tests with the administration of antidepressants (or tranquilizers) and psychostimulators (or nootropic agents), and control examination of mental functions prior to and after drug administration.


Environmental degradation of polyacrylamides. 1. Effects of artificial environmental conditions: temperature, light, and pH.
A polyacrylamide thickening agent (PATA) was formulated at four concentrations in distilled-deionized water, without and with a glyphosate-surfactant herbicide (GH). Over a 6-week period, these mixtures were exposed to various controlled temperature and light conditions. Acrylamide concentration, ammonium concentration, and pH were measured at weekly intervals to assess the degradation of polyacrylamide and acrylamide. Satellite studies were conducted to examine the effect of altered pH on solutions of PATA (i.e., does pH promote polyacrylamide depolymerization?) and GH binding to amine groups (i.e., protection from degradation). The results of these studies suggest that polyacrylamide can degrade to acrylamide by thermal and photolytic effects, that changes in pH do not promote the depolymerization of polyacrylamide, and that GH does protect polyacrylamide and acrylamide from environmental degradation. Statistically there was no linear correlation between the various parameters measured.


Growth and shape-ordering of iron nanostructures on Au single crystalline electrodes in an ionic liquid: a paradigm of magnetostatic coupling.
Fe electrodeposition on Au(111) and Au(100) in BMIBF(4) ionic liquid is found to form hitherto unreported shape-ordered nanoscale morphologies of pseudorods and pseudosquare rings, respectively, both composed of grains of 4-7 nm. The manner of growth of the square rings is a ring-on-ring structure with enlarging side length and slightly protruding four corners. The generality of the growth mechanism is verified by the formation of almost exactly the same shape-ordered Fe nanostructures on Pt, i.e., pseudorod structure on Pt(111) and pseudosquare rings Pt(100). These structures are explained within the framework of magnetostatic interactions of spontaneously magnetized grains under crystallographic constraint of the substrate surface, which result in an antiparallel arrangement in magnetization of the grains at pseudorods and magnetic flux closure at the pseudosquare rings. The closed magnetic flux further leads to magnetic field-enhanced growth at the four corners and the outer peripheries of the pseudosquare rings. The observed shape-ordering of the Fe thin film serves as a paradigm of magnetostatic coupling, in which the roles of ionic liquid as surfactant and magnetic media may not be underestimated. The present work adds a new dimension to electrodeposition in ionic liquid, by which new magnetic film structures may be expected.


Image Quality and Activity Optimization in Oncologic 18F-FDG PET Using the Digital Biograph Vision PET/CT System.
The first Biograph Vision PET/CT system (Siemens Healthineers) was installed at the University Medical Center Groningen. Improved performance of this system could allow for a reduction in activity administration or scan duration. This study evaluated the effects of reduced scan duration in oncologic 18F-FDG PET imaging on quantitative and subjective imaging parameters and its influence on clinical image interpretation. Methods: Patients referred for a clinical PET/CT scan were enrolled in this study, received a weight-based 18F-FDG injected activity, and underwent list-mode PET acquisition at 180 s per bed position (s/bp). Acquired PET data were reconstructed using the vendor-recommended clinical reconstruction protocol (hereafter referred to as "clinical"), using the clinical protocol with additional 2-mm gaussian filtering (hereafter referred to as "clinical+G2"), and-in conformance with European Association of Nuclear Medicine Research Ltd. (EARL) specifications-using different scan durations per bed position (180, 120, 60, 30, and 10 s). Reconstructed images were quantitatively assessed for comparison of SUVs and noise. In addition, clinically reconstructed images were qualitatively evaluated by 3 nuclear medicine physicians. Results: In total, 30 oncologic patients (22 men, 8 women; age: 48-88 y [range], 67 ± 9.6 y [mean ± SD]) received a single weight-based (3 MBq/kg) 18F-FDG injected activity (weight: 45-123 kg [range], 81 ± 15 kg [mean ± SD]; activity: 135-380 MBq [range], 241 ± 47.3 MBq [mean ± SD]). Significant differences in lesion SUVmax were found between the 180-s/bp images and the 30- and 10-s/bp images reconstructed using the clinical protocols, whereas no differences were found in lesion SUVpeak EARL-compliant images did not show differences in lesion SUVmax or SUVpeak between scan durations. Quantitative parameters showed minimal deviation (∼5%) in the 60-s/bp images. Therefore, further subjective image quality assessment was conducted using the 60-s/bp images. Qualitative assessment revealed the influence of personal preference on physicians' willingness to adopt the 60-s/bp images in clinical practice. Although quantitative PET parameters differed minimally, an increase in noise was observed. Conclusion: With the Biograph Vision PET/CT system for oncologic 18F-FDG imaging, scan duration or activity administration could be reduced by a factor of 3 or more with the use of the clinical+G2 or the EARL-compliant reconstruction protocol.


Teaching how to cite scientific articles: a study of citation deviation in citing "multi-authored papers" by top authors from Iranian Universities of Medical Sciences in 2017.
The present study aims at teaching the right citation models in scientific publications by top authors from Iranian universities of medical sciences in 2017, considering the relationship between moral development and self-control variables and model of "citation to multi-authored papers" in these articles. Multi-authorship increases the amount of self-citation and also increases the likelihood of being cited by others. This study is of the applied scientometrics and correlation (model presentation) studies type. The research population in the first part of this study includes all the authors who had an H-index of 10 or more in the scientific databases of 2017. The sample size at this stage was 110, selected by systematic random sampling. The collected data were analyzed using SPSS 16.0. The results of regression analysis based on the concurrent method indicated that the regression model is significant. The coefficient of determination is 0.096 and the F ratio is 5.650, which are significant at the level of p <0.001. In this regression model, the variables of the level of moral development (β = -5.801, p <0.001) and self-control (p < 0.001, β = -0.253) have significant predictive power and can be considered as predictors of behavioral modeling in citing "multi-authored papers". Based on the results, teaching how to avoid blindly citing the "multi-authored papers", which is regarded as a kind of "citation deviation", can, to some extent, lead to strengthening citation indexes.


Quantitative and integrative proteome analysis of peripheral nerve myelin identifies novel myelin proteins and candidate neuropathy loci.
Peripheral nerve myelin facilitates rapid impulse conduction and normal motor and sensory functions. Many aspects of myelin biogenesis, glia-axonal interactions, and nerve homeostasis are poorly understood at the molecular level. We therefore hypothesized that only a fraction of all relevant myelin proteins has been identified so far. Combining gel-based and gel-free proteomic approaches, we identified 545 proteins in purified mouse sciatic nerve myelin, including 36 previously known myelin constituents. By mass spectrometric quantification, the predominant P0, periaxin, and myelin basic protein constitute 21, 16, and 8% of the total myelin protein, respectively, suggesting that their relative abundance was previously misestimated due to technical limitations regarding protein separation and visualization. Focusing on tetraspan-transmembrane proteins, we validated novel myelin constituents using immuno-based methods. Bioinformatic comparison with mRNA-abundance profiles allowed the categorization in functional groups coregulated during myelin biogenesis and maturation. By differential myelin proteome analysis, we found that the abundance of septin 9, the protein affected in hereditary neuralgic amyotrophy, is strongly increased in a novel mouse model of demyelinating neuropathy caused by the loss of prion protein. Finally, the systematic comparison of our compendium with the positions of human disease loci allowed us to identify several candidate genes for hereditary demyelinating neuropathies. These results illustrate how the integration of unbiased proteome, transcriptome, and genome data can contribute to a molecular dissection of the biogenesis, cell biology, metabolism, and pathology of myelin.


Genetic characterization of Puumala hantavirus strains from Belgium: evidence for a distinct phylogenetic lineage.
Puumala hantavirus (PUUV) sequences were recovered from red bank voles (Clethrionomys glareolus) trapped between 1996 and 1998 in four localities of southern Belgium: Thuin, Montbliart, Momignies and Couvin. In addition, three PUUV isolates originating from bank voles trapped in the 1980s in southern (Montbliart) and northern (Turnhout) Belgium were genetically characterized. Analysis of the complete S and partial M segment sequences showed that the Belgian PUUV strains constitute a genetic lineage, distinct from other known PUUV lineages from Europe and Japan. This lineage also includes a wild strain (Cg-Erft) originating from a neighbouring area of Germany. Within the Belgian lineage, geographical clustering of genetic variants was observed. In the Montbliart site, the range of diversity between the most temporally distant strains (from 1986 and 1996-1998) was higher than between those from 1996 and 1998, suggesting slight genetic drift via accumulation of neutral or quasi-neutral substitutions with time.


Production of functional platelets by differentiated embryonic stem (ES) cells in vitro.
Megakaryocytes and functional platelets were generated in vitro from murine embryonic stem (ES) cells with the use of a coculture system with stromal cells. Two morphologically distinctive megakaryocytes were observed sequentially. Small megakaryocytes rapidly produced proplatelets on day 8 of the differentiation, and large hyperploid megakaryocytes developed after day 12, suggesting primitive and definitive megakaryopoiesis. Two waves of platelet production were consistently observed in the culture medium. A larger number of platelets was produced in the second wave; 104 ES cells produced up to 108 platelets. By transmission electron microscopy, platelets from the first wave were relatively rounder with a limited number of granules, but platelets from the second wave were discoid shaped with well-developed granules that were indistinguishable from peripheral blood platelets. ES-derived platelets were functional since they bound fibrinogen, formed aggregates, expressed P-selectin upon stimulation, and fully spread on immobilized fibrinogen. These results show the potential utility of ES-derived platelets for clinical applications. Furthermore, production of gene-transferred platelets was achieved by differentiating ES cells that were transfected with genes of interest. Overexpression of the cytoplasmic domain of integrin beta3 in the ES-derived platelets prevented the activation of alphaIIbbeta3, demonstrating that this system will facilitate functional platelet studies.


Grayscale level connectivity: theory and applications.
A novel notion of connectivity for grayscale images is introduced, defined by means of a binary connectivity assigned at image-level sets. In this framework, a grayscale image is connected if all level sets below a prespecified threshold are connected. The proposed notion is referred to as grayscale level connectivity and includes, as special cases, other well-known notions of grayscale connectivity, such as fuzzy grayscale connectivity and grayscale blobs. In contrast to those approaches, the present framework does not require all image-level sets to be connected. Moreover, a connected grayscale object may contain more than one regional maximum. Grayscale level connectivity is studied in the rigorous framework of connectivity classes. The use of grayscale level connectivity in image analysis applications, such as object extraction, image segmentation, object-based filtering, and hierarchical image representation, is discussed and illustrated.


T5 Exonuclease Hydrolysis of Hepatitis B Virus Replicative Intermediates Allows Reliable Quantification and Fast Drug Efficacy Testing of Covalently Closed Circular DNA by PCR.
Chronic infection with the human hepatitis B virus (HBV) is a major health problem. Virus persistence requires the establishment and maintenance of covalently closed circular DNA (cccDNA), the episomal virus template in the nucleus of infected hepatocytes. Compared to replicative DNA intermediates (relaxed circular DNA [rcDNA]), copy numbers of cccDNA in infected hepatocytes are low. Accordingly, accurate analyses of cccDNA require enrichment of nuclear fractions and Southern blotting or selective quantitative PCR (qPCR) methods allowing discrimination of cccDNA and rcDNA. In this report, we analyzed cccDNA-specific primer pairs for their ability to amplify cccDNA selectively. Using mixtures of defined forms of HBV and genomic DNA, we determined the potential of different nucleases for targeted digestion of the open/relaxed circular DNA forms in the absence and presence of genomic DNA without affecting cccDNA. We found that the combination of T5 exonuclease with a primer set amplifying an approximately 1-kb fragment permits reliable quantification of cccDNA without the requirement of prior nucleus enrichment or Hirt extraction. We tested this method in four different in vitro infection systems and quantified cccDNA copy numbers at increasing multiplicities of inoculated genome equivalents. We further analyzed the kinetics of cccDNA formation and the effect of drugs (interferon, entry inhibitors, and capsid inhibitors) on cccDNA. Our method allows reliable cccDNA quantification at early stages of infection in the presence of a high excess of input virus and replicative intermediates and is thereby suitable for drug screening and investigation of cccDNA formation and maintenance.IMPORTANCE cccDNA elimination is a major goal in future curative regimens for chronic HBV patients. However, PCR-based assays for cccDNA quantification show a principally constrained specificity when high levels of input virus or replicative intermediates are present. Here, we characterized T5 exonuclease as a suitable enzyme for medium-throughput in vitro assays that preserves cccDNA but efficiently removes rcDNA prior to PCR-based quantification. We compared T5 exonuclease with the previously described exonuclease III and showed that both nucleases are suitable for reliable quantification of cccDNA by PCR. We substantiated the applicability of our method through examination of early cccDNA formation and stable accumulation in several in vitro infection models and analyzed cccDNA stability after administration of anti-HBV drugs. Our results support the use of T5 exonuclease for fast and convenient rcDNA removal, especially for early cccDNA quantification and rapid drug testing in in vitro studies.


Normal canine brain maturation at magnetic resonance imaging.
The normal neonatal canine brain exhibits marked differences from that of the mature brain. With development into adulthood, there is a decrease in relative water content and progressive myelination; these changes are observable with magnetic resonance imaging (MRI) and are characterized by a repeatable and predictable time course. We characterized these developmental changes on common MRI sequences and identified clinically useful milestones of transition. To accomplish this, 17 normal dogs underwent MRI of the brain at various times after birth from 1 to 36 weeks. Sequences acquired were T1-weighted (T1W), T2-weighted (T2W), fluid attenuated inversion recovery, short tau inversion recovery, and diffusion weighted imaging sequences. The images were assessed subjectively for gray and white matter relative signal intensity and results correlated with histologic findings. The development of the neonatal canine brain follows a pattern that qualitatively matches that observed in humans, and which can be characterized adequately on T1W and T2W images. At birth, the relative gray matter to white matter signal intensity of the cortex is reversed from that of the adult with an isointense transition at 3-4 weeks on T1W and 4-8 weeks on T2W images. This is followed by the expected mature gray matter to white matter relative intensity that undergoes continued development to a mostly adult appearance by 16 weeks. On the fluid attenuated inversion recovery sequence, the cortical gray and white matter exhibit an additional signal intensity reversal during the juvenile period that is due to the initial high relative water content at the subcortical white matter, with its marked T1 relaxation effect.