Quorum-dependent expression of rsmX and rsmY, small non-coding RNAs, in Pseudomonas syringae.
Pseudomonas syringae pathovars are known to produce N-acyl-homoserine lactones (AHL) as quorum-sensing molecules. However, many isolates, including P. syringae pv. tomato DC3000 (PtoDC3000), do not produce them. In P. syringae, psyI, which encodes an AHL synthase, and psyR, which encodes the transcription factor PsyR required for activation of psyI, are convergently transcribed. In P. amygdali pv. tabaci 6605 (Pta6605), there is one nucleotide between the stop codons of both psyI and psyR. However, the canonical stop codon for psyI in PtoDC3000 was converted to the cysteine codon by one nucleotide deletion, and 23 additional amino acids extended it to a C-terminal end. This resulted in overlapping of the open reading frame (ORF) for psyI and psyR. On the other hand, stop codons in the psyR ORF of P. syringae 7 isolates, including pv. phaseolicola and pv. glycinea, were found. These results indicate that many pathovars of P. syringae have genetically lost AHL production ability by the mutation of their responsible genes. To examine whether PtoDC3000 modulates the gene expression profile in a population-dependent manner, we carried out microarray analysis using RNAs prepared from low- and high-density cells. We found the expressions of rsmX and rsmY remarkably activated in high-density cells. The activated expressions of rsmX and rsmY were confirmed by Northern blot hybridization, but these expressions were abolished in a ΔgacA mutant of Pta6605. These results indicate that regardless of the ability to produce AHL, P. syringae regulates expression of the small noncoding RNAs rsmX/Y by currently unknown quorum-sensing molecules.


Separation of resistance to antitumor diarylsulfonylurea agents from collateral sensitivity to mitochondrial toxins.
Compared with parental GC3/c1 human colon adenocarcinoma cells, which are diarylsulfonylurea (DSU)-sensitive cells, the DSU-resistant clone LYC5 demonstrates 4.2-, 12.8-, and 5.3- fold increase in sensitivity to the mitochondrial toxins rotenone, antimycin, and oligomycin, respectively. Studies with hybrids formed by fusion of parental GC3/c1 cells with LYC5 cells have indicated that resistance to antitumor DSUs and collateral sensitivity to mitochondrial toxins are recessive and therefore potentially linked. To examine this, we transfected a cDNA library from GC3/c1 cells, constructed in pcDNA3, into LYC5 cells. G418-resistant colonies were selected and further selected in a single step for resistance to rotenone (100 nm). Individual colonies (designated T5LR) were expanded and tested for sensitivity to mitochondrial toxins, antitumor DSU agents (LY195779 and LY186391) that demonstrate a 45-50-fold differential potency against GC3/c1, LYC5 cells, and the antimitotic agent vincristine. Results demonstrate that resistance to mitochondrial toxins rotenone, antimycin, and oligomycin can be transferred without conferring a DSU-sensitive phenotype. Furthermore, in T5LR clones, resistance to mitochondrial toxins was not associated with increased resistance to vincristine or increased P-glycoprotein expression, supporting the contention that resistance to these agents is independent of P-glycoprotein. Southern blot analysis of T5LR clones demonstrated unique integration sites for the neomycin phosphotransferase gene into genomic DNA in clones 4 and 9, indicating independent derivation. Analysis of clones 4, 6, and 9 with use of polymerase chain reaction demonstrated a cDNA insert of approximately 1.0 kilobase.


Protofibrillar intermediates of amyloid beta-protein induce acute electrophysiological changes and progressive neurotoxicity in cortical neurons.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder that is thought to be caused in part by the age-related accumulation of amyloid beta-protein (Abeta). The presence of neuritic plaques containing abundant Abeta-derived amyloid fibrils in AD brain tissue supports the concept that fibril accumulation per se underlies neuronal dysfunction in AD. Recent observations have begun to challenge this assumption by suggesting that earlier Abeta assemblies formed during the process of fibrillogenesis may also play a role in AD pathogenesis. Here, we present the novel finding that protofibrils (PF), metastable intermediates in amyloid fibril formation, can alter the electrical activity of neurons and cause neuronal loss. Both low molecular weight Abeta (LMW Abeta) and PF reproducibly induced toxicity in mixed brain cultures in a time- and concentration-dependent manner. No increase in fibril formation during the course of the experiments was observed by either Congo red binding or electron microscopy, suggesting that the neurotoxicity of LMW Abeta and PF cannot be explained by conversion to fibrils. Importantly, protofibrils, but not LMW Abeta, produced a rapid increase in EPSPs, action potentials, and membrane depolarizations. These data suggest that PF have inherent biological activity similar to that of mature fibrils. Our results raise the possibility that the preclinical and early clinical progression of AD is driven in part by the accumulation of specific Abeta assembly intermediates formed during the process of fibrillogenesis.


Nighttime Chemistry and Morning Isoprene Can Drive Urban Ozone Downwind of a Major Deciduous Forest.
Isoprene is the predominant non-methane volatile organic compound emitted to the atmosphere and shapes tropospheric composition and biogeochemistry through its effects on ozone, other oxidants, aerosols, and the nitrogen cycle. Isoprene is emitted naturally by vegetation during daytime, when its photo-oxidation is rapid, and in the presence of nitrogen oxides (NOx) produces ozone and degrades air quality in polluted regions. Here, we show for a city downwind of an isoprene-emitting forest (St. Louis, MO) that isoprene actually peaks at night; ambient levels then endure, owing to low nighttime OH radical concentrations. Nocturnal chemistry controls the fate of that isoprene and the likelihood of a high-ozone episode the following day. When nitrate (NO3) radicals are suppressed, high isoprene persists through the night, providing photochemical fuel upon daybreak and leading to a dramatic late-morning ozone peak. On nights with significant NO3, isoprene is removed before dawn; days with low morning isoprene then have lower ozone with a more typical afternoon peak. This biogenic-anthropogenic coupling expands the daily high-ozone window and likely has an opposite O3-NOx response to what would otherwise be expected, with implications for exposure and air-quality management in cities that, like St. Louis, are downwind of major isoprene-emitting forests.


Infanticide by males in a spider with suicidal maternal care, Stegodyphus lineatus (Eresidae)
Infanticide by males in species with high maternal investment may be the most extreme example of a reproductive conflict of interests between males and females. The subsocial spider Stegodyphus lineatus is semelparous, and the offspring eat their mother. Male and female maturation overlap, such that late maturing males encounter females that are already guarding the single clutch. A late male may try to remove the existing egg sac, thereby forcing the female to replace the clutch and use his sperm to fertilize at least some of the eggs. Males were responsible for 33% of all the egg losses in a population of 278 females with eggs. By losing her clutch, the female loses time equivalent to the age of the lost clutch plus the time it takes to replace it. The earlier in the season a female lost a clutch and the younger the eggs were, the longer was the interval between clutch loss and clutch replacement. Females that lost their egg sacs by infanticide replaced them after a significantly longer time than females that lost their egg sacs from other causes. The time loss of 18.5 days on average corresponds to a 23% reduction in female survival probability. In addition to reduced survival, females suffered a fecundity cost: replacement clutches were smaller than the original clutches and the reduction in egg number was greater with increasing intervals between the first and second clutch. Females defended their egg sacs aggressively against males. In field tests, the outcome of fights between males and females was determined by the relative size difference of the contestants. The mating system of S. lineatus provides an example of a male mating strategy that evolved despite being very costly for the females.


Differential activation mechanisms of Erk-1/2 and p70(S6K) by glucose in pancreatic beta-cells.
Glucose can activate the mitogen-activated kinases, Erk-1/2, and the ribosomal-S6 kinase, p70(S6K), in beta-cells, contributing to an increase in mitogenesis. However, the signaling mechanism by which glucose induces Erk-1/2 and p70(S6K) phosphorylation activation is undefined. Increased glucose metabolism increases [Ca(2+)](i) and [cAMP], and it was investigated if these secondary signals were linked to glucose-induced Erk-1/2 and p70(S6K) activation in pancreatic beta-cells. Blocking Ca(2+) influx with verapamil, or inhibiting protein kinase A (PKA) with H89, prevented glucose-induced Erk-1/2 phosphorylation. Increasing cAMP levels by GLP-1 potentiated glucose-induced Erk-1/2 phosphorylation via PKA activation. Elevation of [Ca(2+)](i) by glyburide potentiated Erk-1/2 phosphorylation, which was also inhibited by H89, suggesting increased [Ca(2+)](i) preceded PKA for glucose-induced Erk-1/2 activation. Adenoviral-mediated expression of dominant negative Ras in INS-1 cells decreased IGF-1-induced Erk-1/2 phosphorylation but had no effect on that by glucose. Collectively, our study indicates that a glucose-induced rise in [Ca(2+)](i) leads to cAMP-induced activation of PKA that acts downstream of Ras and upstream of the MAP/Erk kinase, MEK, to mediate Erk-1/2 phosphorylation via phosphorylation activation of Raf-1. In contrast, glucose-induced p70(S6K) activation, in the same beta-cells, was mediated by a distinct signaling pathway independent of Ca(2+)/cAMP, most likely via mTOR-kinase acting as an "ATP-sensor."


Characterisation of regulatory sequences at the Epstein-Barr virus BamHI W promoter.
Epstein-Barr virus, a human gammaherpesvirus, possesses a unique set of latent genes whose constitutive expression in B cells leads to cell growth transformation. The initiation of this growth transforming infection depends on a viral promoter in BamHI W (Wp) whose regulation is poorly understood. Using Wp reporter constructs in in vitro transfection assays, we found that Wp was 11- to 190-fold more active in B cell than in non-B cell lines and that three regions of the promoter (termed UAS1, UAS2, and UAS3) contributed to transcriptional activation. The upstream regions UAS3 (-1168 to -440) and UAS2 (-352 to -264) both functioned in a cell lineage-independent manner and were together responsible for the bulk of Wp activity in non-B cells; mutational analysis indicated the importance of a YY1 binding site in UAS2 in that context. By contrast, UAS1 (-140 to -87) was B cell specific and was the key determinant of the promoter's increased activity in B cell lines. Mutational analysis of UAS1 sequences combined with in vitro bandshift assays revealed the presence of three binding sites for cellular factors in this region. When mutations that abolished factor binding in bandshift assays were introduced into a Wp reporter construct, the loss of any one of the three UAS1 binding sites was sufficient to reduce promoter activity by 10- to 30-fold in B cells. From sequence analysis, two of these appear to be novel transcription factor binding sites, whereas the third was identified as a cyclic AMP response element (CRE). Our data indicate that this CRE interacts with CREB and ATF1 proteins present in B cell nuclear extracts and that this interaction is important for Wp activity.


Repeated whisker stimulation evokes invariant neuronal responses in the dorsolateral striatum of anesthetized rats: a potential correlate of sensorimotor habits.
The dorsolateral striatum (DLS) receives extensive projections from primary somatosensory cortex (SI), but very few studies have used somesthetic stimulation to characterize the sensory coding properties of DLS neurons. In this study, we used computer-controlled whisker deflections to characterize the extracellular responses of DLS neurons in rats lightly anesthetized with isoflurane. When multiple whiskers were synchronously deflected by rapid back-and-forth movements, whisker-sensitive neurons in the DLS responded to both directions of movement. The latency and magnitude of these neuronal responses displayed very little variation with changes in the rate (2, 5, or 8 Hz) of whisker stimulation. Simultaneous recordings in SI barrel cortex and the DLS revealed important distinctions in the neuronal responses of these serially connected brain regions. In contrast to DLS neurons, SI neurons were activated by the initial deflection of the whiskers but did not respond when the whiskers moved back to their original position. As the rate of whisker stimulation increased, SI responsiveness declined, and the latencies of the responses increased. In fact, when whiskers were deflected at 5 or 8 Hz, many neurons in the DLS responded before the SI neurons. These results and earlier anatomic findings suggest that a component of the sensory-induced response in the DLS is mediated by inputs from the thalamus. Furthermore, the lack of sensory adaptation in the DLS may represent a critical part of the neural mechanism by which the DLS encodes stimulus-response associations that trigger motor habits and other stimulus-evoked behaviors that are not contingent on rewarded outcomes.


Iatrogenic perforation of the left heart during placement of a chest drain.
Chest drain placement is a standard procedure for treating pneumothorax and pleural effusions and has a low complication rate. It is a safe and efficient procedure if image guidance is used. If the anatomic orientation is hampered and neither air nor fluids can be initially aspirated, more complex imaging than a chest x-ray is indicated to avoid major complications. We report the case of an 88-year-old male patient suffering from chronic heart failure who was admitted to another hospital following acute cardiac decompensation. Because of dyspnea with voluminous bilateral effusions, an attempt was made to drain the left pleural cavity. A malposition of the chest drain was suspected because blood was initially draining from the catheter. The hemodynamically stable patient was referred to our university hospital, where computed tomography of the chest revealed the location of the intercostal drain. The drain had perforated the left ventricle, run through the mitral valve and exited the left atrium via a pulmonary vein, ending in the middle lobe. The patient was brought to the surgical theater, where cardiac surgeons performed a left anterolateral thoracotomy and extracted the drain successfully. Three days later, the patient was discharged from our hospital in a good general condition.


Cardiovascular consequences of sleep apnea: II-Cardiovascular mechanisms.
Obstructive sleep apnea (OSA) is a common disorder with serious cardiovascular consequences. The pathogenesis in this context is likely to be multifactorial process including large negative swings in intrathoracic pressure, intermittent hypoxemia and hypercapnia, increased sympathetic nervous system activity, vascular endothelial dysfunction, oxidative stress, systemic inflammation, excessive platelet activation as well as metabolic dysregulation. Although there is scientific support for a considerable impact of OSA on vascular structure and function, it is likely that development of cardiovascular diseases is determined by multiple genotypic and phenotypic factors. The current article focuses on the available research evidence addressing the cardiovascular mechanisms in this context.


Optimal Control Design of Impulsive SQEIAR Epidemic Models with Application to COVID-19.
This paper presents a SEIAR-type model considering quarantined individuals (Q), called SQEIAR model. The dynamic of SQEIAR model is defined by six ordinary differential equations that describe the numbers of Susceptible, Quarantined, Exposed, Infected, Asymptomatic, and Recovered individuals. The goal of this paper is to reduce the size of susceptible, infected, exposed and asymptomatic groups to consequently eradicate the infection by using two actions: the quarantine and the treatment of infected people. To reach this purpose, optimal control theory is presented to control the epidemic model over free terminal optimal time control with an optimal cost. Pontryagin's maximum principle is used to characterize the optimal controls and the optimal final time. Also, an impulsive epidemic model of SQEIAR is considered to deal with the potential suddenly increased in population caused by immigration or travel. Since this model is suitable to describe the COVID-19 pandemic, especial attention is devoted to this case. Thus, numerical simulations are given to prove the accuracy of the theoretical claims and applied to the particular data of this infection. Moreover, numerical computations of the COVID-19 are compared with diseases like Ebola and Influenza. In addition, the controller is evaluated with system parameters identified by using actual data of China. Finally, the controller tuned with the estimated parameters of the Chinese data is applied to the actual data of Spain to compare the quarantine and treatment policies in both countries.


Microstructural white matter changes following gait training with Hybrid Assistive Limb initiated within 1 week of stroke onset.
The early initiation of robot-assisted gait training in patients with acute stroke could promote neuroplasticity. The aim of this study was to clarify the microstructural changes of white matter associated with gait training using Hybrid Assistive Limb (HAL) by diffusion tensor imaging (DTI). Patients with first-ever stroke and requiring a walking aid started gait training within 1 week of stroke onset. The patients were quasi-randomly assigned either to the conventional physical therapy (CPT) group or gait training using HAL (HAL) group. Motor function and DTI were examined at baseline and after 3-5 months. Voxel-based statistical analyses of fractional anisotropy (FA) images were performed using diffusion metric voxel-wise analyses. Volume of interest (VOI)-based analyses were used to assess changes in FA (ΔFA). Twenty-seven patients (17 in the CPT group and 10 in the HAL group) completed the study. There were improvements in motor function and independency in the CPT and HAL groups (p < .001). Compared to baseline, there were decreases in FA in the ipsi-lesional cerebral peduncle in the CPT group (p < .001) and increases in the contra-lesional rostrum of the corpus callosum in the HAL group (p < .001) at the second assessment, consistent with the mean ΔFA in each group from VOI analysis (CPT/HAL: cerebral peduncle, -0.066/-0.027, p = .027; corpus callosum, 0.002/0.042, p < .001). Gait training using HAL initiated within 1 week after stroke onset facilitated the recovery of inter-hemispheric communication and prevented the progression of Wallerian degeneration of the affected pyramidal tract.


Height-for-age and weight-for-age growth charts for Pakistani infants under six months: derived from a novel case selection method using multiple indicator cluster survey data.
In the past two decades, there has been a growing recognition of the need to establish indigenous standards or reference growth charts, particularly following the WHO multicenter growth study in 2006. The availability of accurate and reliable growth charts is crucial for monitoring child health. The choice of an appropriate model for constructing growth charts depends on various data characteristics, including the distribution's tails and peak. While Pakistan has reported some reference growth charts, there is a notable absence of indigenous charts for children under two years of age, especially for infants aged 0-6 months who are exclusively breastfed. Additionally, acquiring data poses a significant challenge, particularly for low-income countries, as it demands substantial resources such as finances, time, and expertise. The Multiple Indicator Cluster Survey (MICS) constitutes a large-scale national survey conducted periodically in low-income countries under the auspices of UNICEF. In this study, we propose methods for generating selection variables utilizing the "Novel Case Selection Method," as previously published. Further our approach enables to select and fit appropriate model to the MICS data, selected, and to develop the standard growth charts. Out of the 11,478 children under 6 months of age included in MICS-6 (Pakistan), 3,655 children (1,831 males and 1,824 females) met the specified criteria and were selected using the "Novel Case Selection Method". The sample was distributed across provinces as follows: 841 (23.0%) from KPK, 1,464 (40.1%) from Punjab, 819 (22.4%) from Sindh, and 531 (14.5%) from Balochistan. This sample encompassed both rural (76.4%) and urban (23.6%) populations. Following data cleaning and outlier removal, a total of 3,540 records for weight (1,768 males and 1,772 females) and 3,515 records for height (1,759 males and 1,756 females) were ultimately available for the development of standard charts. The Bayesian Information Criterion (BIC) was employed to determine the optimal degrees of freedom for L, M, and S using RefCurv_0.4.2. Three families within the gamlss class-namely, Box Cox Cole and Green (BCCG), Box Cox T (BCT), and Box Cox Power Exponential (BCPE)-were applied, each with three smoothing techniques: penalized splines (ps), cubic splines (cs), and polynomial splines (poly). The best-fitted model was selected from these nine combinations based on the Akaike Information Criteria. The Novel Case Selection Method yielded 3655 cases as per criteria. After cleaning the data, this method lead to selection of 3540 children for "weight for age" (W/A) and 3515 children for "height for age" (H/A). The "BCPE" family and "ps" as smoothing method proved to be best on AIC for all four curves, i.e. the W/A male, W/A female, H/A male, and H/A female. The optimum selected degrees of freedom for the curve "W/A", for both genders were (M = 1, L = 0, S = 0). The optimum degrees of freedom for H/A male were again (M = 1, L = 0, S = 0), but for females the selected degrees of freedom were (M = 1, L = 1, S = 1). The indigenous fitted standard curves for Pakistan were on lower trajectory in comparison to WHO standards. This study uses the Novel Case Selection Method with introduced algorithms to construct tailored growth charts for lower and middle-income countries. Leveraging extensive MICS data, the methodology ensures representative national samples. The resulting charts hold practical value and await validation from established data sources, offering valuable tools for policy makers and clinicians in diverse global contexts.


Late onset multiple sclerosis: concerns in aging patients.
Late onset multiple sclerosis (LOMS) is when the first symptom starts after 50 years of age, representing 4.5% of multiple sclerosis (MS) patients. This study describes the clinical characteristics of patients with LOMS followed at a specialized MS center in São Paulo. Data was obtained from medical records of 742 patients with MS. The LOMS frequency was 4.18%, median age at onset was 54 years and the predominant disease course was primary progressive (64.3%). The patients reached the disability landmarks of EDSS grades 3.0, 6.0 and 7.0 in the following proportion and time: EDSS 3.0: 77.42% of patients in 3.7 years; EDSS 6.0: 58.06% in 5.1 years and EDSS 7.0: 32.26% in 5.7 years. The comparative analysis with a matched control group of patients with early onset MS showed that late onset, associated with a progressive course, were predictors of reaching EDSS 3.0 and 6.0 in a shorter time.


Direct detection of transcription factors in cotyledons during seedling development using sensitive silicon-substrate photonic crystal protein arrays.
Transcription factors control important gene networks, altering the expression of a wide variety of genes, including those of agronomic importance, despite often being expressed at low levels. Detecting transcription factor proteins is difficult, because current high-throughput methods may not be sensitive enough. One-dimensional, silicon-substrate photonic crystal (PC) arrays provide an alternative substrate for printing multiplexed protein microarrays that have greater sensitivity through an increased signal-to-noise ratio of the fluorescent signal compared with performing the same assay upon a traditional aminosilanized glass surface. As a model system to test proof of concept of the silicon-substrate PC arrays to directly detect rare proteins in crude plant extracts, we selected representatives of four different transcription factor families (zinc finger GATA, basic helix-loop-helix, BTF3/NAC [for basic transcription factor of the NAC family], and YABBY) that have increasing transcript levels during the stages of seedling cotyledon development. Antibodies to synthetic peptides representing the transcription factors were printed on both glass slides and silicon-substrate PC slides along with antibodies to abundant cotyledon proteins, seed lectin, and Kunitz trypsin inhibitor. The silicon-substrate PC arrays proved more sensitive than those performed on glass slides, detecting rare proteins that were below background on the glass slides. The zinc finger transcription factor was detected on the PC arrays in crude extracts of all stages of the seedling cotyledons, whereas YABBY seemed to be at the lower limit of their sensitivity. Interestingly, the basic helix-loop-helix and NAC proteins showed developmental profiles consistent with their transcript patterns, indicating proof of concept for detecting these low-abundance proteins in crude extracts.


Tryptophan depletion affects the autonomic stress response in generalized social anxiety disorder.
In generalized social anxiety disorder (gSAD), serotonergic dysfunctions are found, as well as abnormalities of the autonomic nervous system (ANS) in basal conditions and of the hypothalamic pituitary adrenal (HPA) axis in response to psychological challenges. These findings raise the question whether these phenomena are interrelated. Therefore we designed a study in which two groups with nine pair wise age and gender matched gSAD patients (total of 10 men and 8 women), who were successfully treated with a selective serotonin reuptake inhibitor (SSRI), underwent a tryptophan depletion challenge (TD) or a placebo condition. A TD procedure temporarily decreases serotonergic neurotransmission. In order to activate the stress system the TD/placebo challenge was combined with a public speaking task. We assessed ANS responses, as measured with the promising new marker salivary alpha-amylase (sAA), and HPA-axis responses, as measured with salivary cortisol. The most important result was that the TD group showed a significant larger sAA response to the public speaking task as compared to the placebo group, reflecting hyperresponsivity of the ANS in this group, whereas no differences were seen in cortisol responses. This suggests that in gSAD there is a vulnerability of the ANS more than the HPA-axis.


Sitosterolemia presenting with severe hypercholesterolemia and intertriginous xanthomas in a breastfed infant: case report and brief review.
Sitosterolemia is an autosomal recessive disorder characterized by increased intestinal absorption of plant sterols. It is caused by mutations in genes encoding ATP-binding cassette, subfamily G5 (ABCG5) or G8 (ABCG8), and clinical features include elevated plant sterol levels, xanthomas, and accelerated atherosclerosis. Although it was originally reported in patients with normolipemic xanthomas, patients with sitosterolemia also hyperabsorb cholesterol, and serum cholesterol levels tend to be elevated. We report an infant with sitosterolemia who presented with severe hypercholesterolemia and intertriginous xanthomas. A 15-month-old Korean girl presented with yellow dermal plaques over flexural areas including the wrist, neck, and gluteal folds, which were consistent with intertriginous xanthomas. The lesions were first noticed at 3 months of age when she was being exclusively breastfed. Her total cholesterol and low-density lipoprotein-cholesterol levels were 675 and 540 mg/dL, respectively. A low-fat/low-cholesterol diet and cholestyramine therapy were introduced. Unexpectedly, her serum cholesterol level decreased dramatically and normalized in 2 months. Cholestyramine was tapered off. The xanthomas also regressed and disappeared by 3 years of age. Gas chromatography-mass spectrometric analysis was performed with serum drawn at 3 years of age when her low-density lipoprotein-cholesterol was 118 mg/dL, which revealed striking elevation of her sitosterol level at 19.36 mg/dL. Direct sequencing for ABCG5 revealed compound heterozygous null mutations c.904+1G>A (p.Met302Asnfs*82) and c.1336C>T(p.Arg446*). Our case suggests that sitosterolemia can present with severe hypercholesterolemia and intertriginous xanthomas. Sitosterolemia should be suspected when a patient with hypercholesterolemia shows unexpectedly good response to dietary modification or bile acid sequestrant therapy.


Central prolactin modulates insulin sensitivity and insulin secretion in diabetic rats.
Prolactin secretion is self-regulating as it acts upon hypothalamic dopaminergic systems which inhibit prolactin release from the anterior pituitary. Circulating prolactin improves glucose homeostasis by increasing insulin action and secretion, but central prolactin effects on glucose homeostasis have not been examined. Here, we determined that chronic central infusion of prolactin modulates insulin resistance and β-cell function and mass in 90% of pancreatectomized diabetic male rats. Diabetic rats were divided into three groups according to the dose of intracerebroventricular infusion of prolactin during 4 weeks: (1) low-dose prolactin (Low-PRL; 0.1 µg/h), (2) high-dose prolactin (High-PRL; 1 µg/h) and (3) vehicle only (cerebrospinal fluid). Nondiabetic rats were centrally infused with the vehicle. Chronic intracerebroventricular infusion of Low-PRL lowered body weight and epididymal fat pads by increasing hypothalamic dopamine levels that reduced serum prolactin levels and potentiated leptin signaling. However, High-PRL slightly exacerbated energy dysregulation, decreased hypothalamic dopamine levels, and elevated serum prolactin levels. Both dosages promoted β-cell mass but in a different manner: Low-PRL decreased β-cell apoptosis, whereas High-PRL increased its proliferation. However, only Low-PRL enhanced first-phase insulin secretion and improved insulin sensitivity at a hyperglycemic state in comparison to the control. Low-PRL also increased glucose infusion rates and decreased hepatic glucose output in hyperinsulinemic states, signifying an improvement in hepatic insulin sensitivity. However, High-PRL exacerbated hepatic insulin resistance compared with the control diabetic rats. In contrast to the exacerbation of insulin resistance caused by High-PRL, Low-PRL may improve energy and glucose metabolism by increasing hypothalamic dopamine levels in diabetic rats.


Endoscopic Ultrasound Guided Gallbladder Interventions: a Review of the Current Literature.
Interventional endoscopic ultrasound (EUS) is a rapidly expanding field with a wide variety of indications, including different drainage procedures and delivery of locoregional treatment mainly for pancreatic solid tumors. Transgastric or transduodenal gallbladder drainage in high-risk patients with acute cholecystitis or biliary decompression in patients with unresectable distal biliary malignant obstruction who failed endoscopic retrograde colangiography is one of the newest areas of EUS-guided intervention. The large-caliber lumen apposing metal stents placed during these procedures allow direct endoscopic gallbladder access and the possibility of performing gallstone treatment or resection of mucosal polyps. The current review presents the indications of endoscopic gallbladder interventions and discusses the results of available studies, foreseeing future potential applications.


Population coverage by HLA class-I restricted cytotoxic T-lymphocyte epitopes.
Vaccination using cytotoxic T-lymphocyte (CTL) epitopes has become a widely used immunization strategy, especially because their structure makes them an attractive alternative to the delivery of whole proteins as immunogens. Nonetheless, their use is limited, in particular because of their specificity, being recognized only by cognate HLA alleles. The potential for immunizing a substantial portion of an ethnically diverse population using a modest number of peptides has been aided by the identification of HLA supertypes. However, the derivation of epitopes is often guided by methods that do not guarantee cross-reactivity, so we consider the feasibility of providing vaccine coverage to a multi-ethnic population under different assumptions. In particular, two large datasets are used to estimate the number of peptides needed to provide > or =90% group-specific coverage of a multiethnic population, when specificity is assumed to be either to a single serologic or molecular type. These assumptions are evaluated utilizing a clinically important epitope repertoire derived from two human cytomegalovirus proteins, and data on the in vitro memory response elicited by these peptides is presented. In summary, our combined theoretical and empiric studies suggest that 90% coverage of some ethnic groups is attainable with 11 uniquely defined HLA-restricted CTL epitopes. The derivation of four or more additional CTL epitopes is needed to attain 90% coverage of Blacks or Asians, the minimally covered groups. Ninety percent coverage of all major ethnic groups in a multi-ethnic population appears feasible without relying on cross-reactivity, but may require two to three times more CTL epitopes than estimated for serologic data, homogenous populations, or HLA alleles grouped as supertypes.


Reoperation rate and risk factors of elective spinal surgery for degenerative spondylolisthesis: minimum 5-year follow-up.
The favorable outcome of surgical treatment for degenerative lumbar spondylolisthesis (DS) is widely recognized, but some patients require reoperation because of complications, such as pseudoarthrosis, persistent pain, infection, and progressive degenerative changes. Among these changes, adjacent segmental disease (ASD) and same segmental disease (SSD) are common reasons for reoperation. However, the relative risks of the various factors and their interactions are unclear. The purpose of this study was to determine the longitudinal reoperation rate after surgery for DS and to assess the incidence and independent risk factors for ASD and SSD. This study is a retrospective consecutive case series of patients with DS who were surgically treated. We assessed 163 consecutive patients who were surgically treated for DS between 2003 and 2008. Individual patients were followed for at least 5 years after the initial surgery. The primary end point was any type of second lumbar surgery. Radiographic measurements and demographic data were reviewed. We compared patients who underwent reoperation with those who did not. Logistic regression analysis was used to determine the relative risk of ASD and SSD in patients surgically treated for DS. Radiographic measurements and demographic data were reviewed. We identified the incidence and risk factors for reoperation, and we performed univariate and multivariate analyses to determine the independent risk factors for revision surgery for SSD and for ASD as the two distinct reasons for the reoperation. Age, gender, etiology, body mass index (BMI), and other radiographic data were analyzed to determine the risk factors for developing SSD and ASD. The average patient age was 65.8 (50-81 years; 73 women and 90 men; mean follow-up, 5.9±1.6 years). Eighty-nine patients had posterior lumbar interbody fusion and 74 had laminotomies. Twenty-two patients had L3-L4 involvement and 141 had L4-L5 involvement. The cumulative reoperation rate was 6.1% at 1 year, 8.5% at 2 years, 15.2% at 3 years, 17.7% at 5 years, and 23.3% (38/163 patients) at the final follow-up. A significantly higher reoperation rate was observed for patients undergoing laminotomy than for patients undergoing posterior lumbar interbody fusion (33.8% vs. 14.4%, p=.01). Eighteen patients (11.0%) had SSD, and 13 patients (8.9%) developed ASD. Higher BMI (obesity) and greater disc height (greater than 10 mm) predicted the occurrence of SSD in the multivariate model (BMI=odds ratio 4.11 [95% confidence interval 1.29-13.11], p=.016; disc height=3.18 [1.03-9.82], p=.044), and gender (male) and facet degeneration (Fujiwara grade greater than 3) predicted the development of ASD in the multivariate model (gender=4.74 [1.09-20.45], p=.037; facet degeneration=6.31 [1.09-36.52], p=.039). The incidence of reoperation in patients surgically treated for DS was 23.2% at a mean time of 5.9 years. A significantly higher incidence of reoperation was observed in patients treated with decompression alone compared with those treated with decompression and fusion. Body mass index and disc height were identified as independent risk factors for SSD, whereas male gender and facet degeneration were identified as independent risk factors for ASD. The results of this comprehensive review will guide spine surgeons in their preoperative planning and in the surgical management of patients with DS, thereby reducing the reoperation rate.


Safety and tolerability of a one-time, in-office administration of 5% povidone-iodine in the treatment of adenoviral conjunctivitis: The Reducing Adenoviral Patient Infected Days (RAPID) study.
To report safety and tolerability of a one-time administration of ophthalmic 5% povidone-iodine (5% PVP-I) in a double-masked randomized trial for the treatment of adenoviral conjunctivitis (Ad-Cs). Of 212 participants screened, 56 eligible participants with red eye symptoms ≤4 days and a positive adenoviral rapid immunoassay were randomized to a one-time administration of ophthalmic 5% PVP-I or preservative free artificial tears (AT). Safety was assessed by corneal fluorescein staining (baseline, immediate post-administration and Day 1) and visual acuity (VA) (baseline and Day 1). Tolerability was assessed using participant-rated overall ocular discomfort (baseline, immediately post-administration and on Day 1. In the 5% PVP-I group, corneal staining increased immediately post-administration but returned to baseline levels by Day 1. There was no change in VA between baseline and Day 1 in either 5% PVP-I or AT groups (p = 0.87). In the 5% PVP-I group, there was no change in participant-rated overall discomfort immediately post-administration (p = 0.78) or on day 1 (p = 0.10) compared to baseline. In the AT group, participant-rated overall discomfort was lower immediately post-administration but returned to baseline levels by Day 1. One adverse event was reported in the 5% PVP-I group on Day 1-2 that was classified as not related to treatment. These results suggest ophthalmic 5% PVP-I used as a one-time treatment is safe and well tolerated by patients with Ad-Cs.


The carbon stock potential of the restored mangrove ecosystem of Pasarbanggi, Rembang, Central Java.
Mangrove ecosystems can absorb significant amounts of carbon and help mitigate climate change. However, their existence continues to be endangered by natural and human forces. Therefore, mangrove restoration is regarded as a crucial component of the global climate change agenda. This study aims to estimate the potential total carbon stock of restored mangrove ecosystems in Pasarbanggi, Rembang, Central Java. The above-below-ground (root) carbon stock was calculated using several published allometric equations. The loss-on-ignition method analyzed leaf litter and sediment carbon stocks. This study estimates the Pasarbanggi mangrove ecosystem's total carbon stock potential at 0.02 × 106 MgC, which is equivalent to the potential CO2 emission of 0.08 × 106 MgCO2e, with up to 65% stored in sediments. This study highlights the critical role of restored mangrove ecosystems on the climate change mitigation agenda by reducing the concentration of atmospheric CO2.


The Effect of Acupuncture on Visual Function in Patients with Congenital and Acquired Nystagmus.
Background: The aim of this study is to examine the short-term effect of visual function following acupuncture treatment in patients with congenital idiopathic nystagmus and acquired nystagmus (CIN and AN). Methods: An observational pilot study on six patients with confirmed diagnosis of nystagmus (three CIN and three AN patients (2♀, 4♂; mean age 42.67; SD ± 20.57 y)), was performed. Acupuncture treatment was done following a standardized protocol applying needle-acupuncture on the body and the ears. The treatment was scheduled with 10 sessions of 30 min duration over five weeks. To assess the effect of the treatment, we performed before, between, and after acupuncture objective measurement of the BCVA (EDTRS charts), contrast vision (CSV-1000, Vector Vision), nystagmography (Compact Integrated Pupillograph), complemented by evaluation questionnaires. A placebo non-acupuncture control group (Nr: 11, 22 eyes; 8♀, 3♂; mean age: 33.34 y (SD ± 7.33 y)) was taken for comparison. Results: The results showed that, following acupuncture treatment, CIN and AN patients showed improvement (SD± mean) in their binocular BCVA (baseline: 0.45 ± 0.36; between: 0.53 ± 0.34 and post-treatment: 0.51 ± 0.28), and in their monocular contrast sensitivity (baseline: 11.29 ± 12.35; between: 11.43 ± 11.45 and post-treatment: 14.0 ± 12.22). The post-/baseline-difference showed a significant improvement in contrast vision and in BCVA for CIN and AN patients, but not for controls (p = 0.029 and p = 0.007, respectively). The effect of the eye showed also, within CIN and AN, significant values for the examined parameters in the post-/baseline difference (p = 0.004 and p ≤ 0.001). Evaluated only binocularly, the respective between-/baseline and post-/baseline difference in the CIN and AN group showed significant values (p < 0.045). Two AN patients reported reduction of oscillations. Among general subjective symptoms, our patients reported reduction of tiredness and headache attacks, improvement of vision, and shorter sleep onset time. Conclusion: The applied acupuncture protocol showed improvement in the visual function of nystagmus patients and thus, in their quality of life. Further studies are mandatory to differentiate which group of nystagmus patients would benefit more from acupuncture.


Myoplasmic [ca], crossbridge phosphorylation and latch in rabbit bladder smooth muscle.
TONIC SMOOTH MUSCLE EXHIBIT THE LATCH PHENOMENON: high force at low myosin regulatory light chains (MRLC) phosphorylation, shortening velocity (Vo), and energy consumption. However, the kinetics of MRLC phosphorylation and cellular activation in phasic smooth muscle are unknown. The present study was to determine whether Ca(2+)-stimulated MRLC phosphorylation could suffice to explain the agonist- or high K(+)-induced contraction in a fast, phasic smooth muscle. We measured myoplasmic [Ca(2+)], MRLC phosphorylation, half-time after step-shortening (a measure of Vo) and contractile stress in rabbit urinary bladder strips. High K(+)-induced contractions were phasic at both 22℃ and 37℃: myoplasmic [Ca(2+)], MRLC phosphorylation, 1/half-time, and contractile stress increased transiently and then all decreased to intermediate values. Carbachol (CCh)-induced contractions exhibited latch at 37℃: stress was maintained at high levels despite decreasing myoplasmic [Ca(2+)], MRLC phosphorylation, and 1/half-time. At 22℃ CCh induced sustained elevations in all parameters. 1/half-time depended on both myoplasmic [Ca(2+)] and MRLC phosphorylation. The steady-state dependence of stress on MRLC phosphorylation was very steep at 37℃ in the CCh- or K(+)-depolarized tissue and reduced temperature flattend the dependence of stress on MRLC phosphorylation compared to 37℃. These data suggest that phasic smooth muscle also exhibits latch behavior and latch is less prominent at lower temperature.


Influence of Match Location, Quality of Opponents, and Match Status on Movement Patterns in Brazilian Professional Football Players.
Aquino, R, Munhoz Martins, GH, Palucci Vieira, LH, and Menezes, RP. Influence of match location, quality of opponents and match status on movement patterns in brazilian professional football players. J Strength Cond Res 31(8): 2155-2161, 2017-The aim of this study was to examine the independent and interactive effects of match location, quality of opponents, and match status on the movement patterns in a professional Brazilian football team. Sixteen matches of the fourth division Brazilian Championship of 2015 were analyzed during the competitive stages (classifier, 8 matches; octave-finals, 2 matches; quarterfinals, 2 matches; semifinals, 2 matches; and finals, 2 matches). A 5-Hz Global Positioning System Sports QSTARZ was used to record the total distance (TD), maximum speed (VMAX), average speed (VAVERAGE), and frequency of high-intensity activities (HIA). The Student's t-test for independent samples showed significantly higher values (p ≤ 0.05) of VMAX, VAVERAGE, and HIA in home matches when compared with away matches. Comparing the quality of opponents, statistically higher values of TD, VMAX, and HIA were found when the team played against strong opponents. Regarding match status, 1-way analysis of variance demonstrated that when the team won presented significantly higher values of TD, VMAX, VAVERAGE, and HIA compared with matches when the team lost. There were no substantial interactive effects of match situational variables on movement patterns. Finally, multiple linear regression showed that the variable quality of opponents has a higher relative contribution to the variance in HIA (19%) than match status (16%) and match location (4%). In particular, the results indicate that physical performance in professional football is influenced by match situational variables, resulting in a change in the team's style of play.


Exploring biochemical pathways for mono-ethylene glycol (MEG) synthesis from synthesis gas.
Mono-ethylene glycol (MEG) is an important petrochemical with widespread use in numerous consumer products. The current industrial MEG-production process relies on non-renewable fossil fuel-based feedstocks, such as petroleum, natural gas, and naphtha; hence, it is useful to explore alternative routes of MEG-synthesis from gases as they might provide a greener and more sustainable alternative to the current production methods. Technologies of synthetic biology and metabolic engineering of microorganisms can be deployed for the expression of new biochemical pathways for MEG-synthesis from gases, provided that such promising alternative routes are first identified. We used the BNICE.ch algorithm to develop novel and previously unknown biological pathways to MEG from synthesis gas by leveraging the Wood-Ljungdahl pathway of carbon fixation of acetogenic bacteria. We developed a set of useful pathway pruning and analysis criteria to systematically assess thousands of pathways generated by BNICE.ch. Published genome-scale models of Moorella thermoacetica and Clostridium ljungdahlii were used to perform the pathway yield calculations and in-depth analyses of seven (7) newly developed biological MEG-producing pathways from gases, including CO2, CO, and H2. These analyses helped identify not only better candidate pathways, but also superior chassis organisms that can be used for metabolic engineering of the candidate pathways. The pathway generation, pruning, and detailed analysis procedures described in this study can also be used to develop biochemical pathways for other commodity chemicals from gaseous substrates.


Proteomic identification of novel targets regulated by the mammalian target of rapamycin pathway during oligodendrocyte differentiation.
Previous work from our laboratory demonstrated that the mammalian target of rapamycin (mTOR) is active during and required for oligodendrocyte progenitor cell (OPC) differentiation. Here, we applied an iTRAQ mass spectrometry-based proteomic approach to identify novel targets of the mTOR pathway during OPC differentiation. Among the 978 proteins identified in this study, 328 (34%) exhibited a greater than 20% change (P < 0.05) in control versus rapamycin-treated cultures following 4 days of differentiation in vitro. Interestingly, 197 (20%) proteins were elevated in rapamycin-treated cultures, while 131 (13%) proteins were downregulated by rapamycin. In support of our previous data, inhibiting mTOR caused a dramatic reduction in the expression of myelin proteins. mTOR also was required for the induction of proteins involved in cholesterol and fatty acid synthesis, as well as the expression of many cytoskeletal proteins, cell signaling components, and nuclear/transcriptional regulators. Of particular interest was the identification of several critical mediators of oligodendrocyte differentiation. Specifically, mTOR activity controls the developmentally programmed upregulation of the prodifferentiation factors Fyn and Quaking, whereas the expression of the differentiation repressor Gpr17 was elevated by mTOR inhibition. These data reveal a distinct signature of mTOR-regulated protein expression during OPC differentiation.


[Method of making a pattern of the surgical wound].
The method of making a pattern of the skin wound defects by means of polyvinylchloride film with a glue layers is described. This method allows one to make rapidly and with the utmost precision the replica of a wound defect of complex configuration located in multiprofile anatomical regions.


[Vasoactive and cardiotonic drugs in the treatment of initial manifestations of cerebrovascular insufficiency in occlusive diseases of major blood vessels of the head].
Patients (n = 118) with initial manifestations of brain blood supply failure induced by stenoses and occlusions of the main vessels of the head were treated by cardio- and vasoactive drugs (corglycon, strophanthin, sulfocamphocaine, euphylline, cavinton, verapamil) under control of the systemic (by Evans blue dilution according to Stewart-Hamilton) and cerebral (by rheoencephalography and ultrasound dopplerography) hemodynamics. As a result of such treatment policy, the number of cases with a complete reverse development of the symptomatology was 16.8% greater than in analogous situations where no therapy with the method proposed was carried out.


Twin boundary profiles with linear-quadratic coupling between order parameters.
A new type of twin boundary was found when two order parameters interact by linear-quadratic coupling QP(2). In this solution, we find that a domain wall consists of two layers in which in one layer both order parameters Q and P are active while in the second layer only Q is active. The adjacent domains are equally asymmetric (Q, P) and (Q, 0) so that one phase could be polar and/or magnetic and contain a ferroelastic strain while the second layer is ferroelastic only without polar or magnetic properties. The two layers represent a stepwise transition between the two domains.We analyze the full phase diagram depending on the coupling constant and anisotropy of the gradient term, and show that in a certain regime the order parameter Q becomes activated only in the interfacial region. A common solution contains kinks and breathers whereby the width of the interface can be very wide in agreement with the first order character of the transition.


Left ventricular hypertrophy is more prevalent in blacks than whites in the general population: the Dallas Heart Study.
Although recent studies have suggested that blacks compared with whites have an increased prevalence of left ventricular hypertrophy, it remains uncertain whether this is true despite adjustment for body composition (fat mass and fat-free mass) and when assessed by cardiac MRI in the general population. The Dallas Heart Study is a population-based study of Dallas County in which 1335 black and 858 white participants 30 to 67 years of age underwent detailed assessment including dual-energy x-ray absorptiometry scan to measure body composition and cardiac MRI. Left ventricular hypertrophy, whether defined by indexation to body surface area (P<0.001), fat-free mass (P=0.002), or height2.7 (P<0.001) was 2- to 3-fold more common in black versus white women. Similar results were seen when comparing black and white men (P<0.001 when left ventricular hypertrophy was indexed to body surface area or height2.7 and P=0.05 when indexed to fat-free mass). Ethnic disparities in left ventricular mass persisted in multivariable models despite adjustment for fat mass, fat-free mass, systolic blood pressure, age, gender, and measures of socioeconomic status. We conclude that blacks compared with whites have increased left ventricular mass and a 2- to 3-fold higher prevalence of left ventricular hypertrophy in the general population, as assessed by cardiac MRI. The ethnic differences in left ventricular mass are independent of differences in body composition.


Synthesis of hepatitis B surface antigen in mammalian cells: expression of the entire gene and the coding region.
We have constructed two simian virus 40 early replacement recombinants that have the coding sequences for hepatitis B virus surface antigen (HBsAg). One construction, LSV-HBsAg, has the coding region for HBsAg but not the portion encoding the putative pre-surface antigen leader. Transformed monkey kidney cells (COS) infected with this recombinant express large quantities of the characteristic partially glycosylated HBsAg molecule, which are assembled into 22-nm particles that appear similar to those produced by human liver cells infected with hepatitis B virus. This result indicates that the pre-surface antigen sequences are not required for the synthesis of HBsAg or its assembly into particulate structures. The second recombinant, LSV-HBpresAg, has the entire surface antigen gene, including the putative promoter and pre-surface antigen region. COS cells infected with this recombinant plasmid produce 40- to 50-fold less HBsAg than those infected with the LSV-HBsAg recombinant plasmid. RNA mapping studies suggest that the transcription of the HBsAg gene is initiated at more than one site, or alternatively, that RNA splicing of transcripts occurs in the pre-surface antigen region.


Detection of co-eluted peptides using database search methods.
Current experimental techniques, especially those applying liquid chromatography mass spectrometry, have made high-throughput proteomic studies possible. The increase in throughput however also raises concerns on the accuracy of identification or quantification. Most experimental procedures select in a given MS scan only a few relatively most intense parent ions, each to be fragmented (MS2) separately, and most other minor co-eluted peptides that have similar chromatographic retention times are ignored and their information lost. We have computationally investigated the possibility of enhancing the information retrieval during a given LC/MS experiment by selecting the two or three most intense parent ions for simultaneous fragmentation. A set of spectra is created via superimposing a number of MS2 spectra, each can be identified by all search methods tested with high confidence, to mimick the spectra of co-eluted peptides. The generated convoluted spectra were used to evaluate the capability of several database search methods - SEQUEST, Mascot, X!Tandem, OMSSA, and RAId_DbS - in identifying true peptides from superimposed spectra of co-eluted peptides. We show that using these simulated spectra, all the database search methods will gain eventually in the number of true peptides identified by using the compound spectra of co-eluted peptides. Reviewed by Vlad Petyuk (nominated by Arcady Mushegian), King Jordan and Shamil Sunyaev. For the full reviews, please go to the Reviewers' comments section.


Regulation of the association of adducin with actin filaments by Rho-associated kinase (Rho-kinase) and myosin phosphatase.
The small GTPase Rho is believed to regulate the actin cytoskeleton and cell adhesion through its specific targets. We previously identified the Rho targets: protein kinase N, Rho-associated kinase (Rho-kinase), and the myosin-binding subunit (MBS) of myosin phosphatase. Here we purified MBS-interacting proteins, identified them as adducin, and found that MBS specifically interacted with adducin in vitro and in vivo. Adducin is a membrane-skeletal protein that promotes the binding of spectrin to actin filaments and is concentrated at the cell-cell contact sites in epithelial cells. We also found that Rho-kinase phosphorylated alpha-adducin in vitro and in vivo and that the phosphorylation of alpha-adducin by Rho-kinase enhanced the interaction of alpha-adducin with actin filaments in vitro. Myosin phosphatase composed of the catalytic subunit and MBS showed phosphatase activity toward alpha-adducin, which was phosphorylated by Rho-kinase. This phosphatase activity was inhibited by the phosphorylation of MBS by Rho-kinase. These results suggest that Rho-kinase and myosin phosphatase regulate the phosphorylation state of adducin downstream of Rho and that the increased phosphorylation of adducin by Rho-kinase causes the interaction of adducin with actin filaments.


Correlations between structural and mechanical properties of human trabecular femur bone.
In this article, the author presents mathematical relationships between the structural and mechanical properties of cancellous human bone tissue obtained by experimental analysis of specimens. Bone tissue material can adjust its internal structure to the existing loading conditions. The mechanical properties affect the structural properties but changes in structural properties likewise cause changes in the mechanical properties of the tissue. In normal tissue, the processes of tissue construction, destruction, and reconstruction are mutually balanced and complementary; if that balance is disturbed, lesions can occur. Therefore, normal bone tissue and pathologically changed tissue (osteoporosis and osteoarthrosis) coming from the area of human femoral head were examined. The structural properties of cancellous tissue specimens were determined non-destructively for three-dimensional reconstructions with the use of modern micro-CT methods. The mechanical properties of the specimens were determined by an uniaxial compression test in three orthogonal directions. Next, in order to specify the compressive strength, a failure test was conducted in the direction perpendicular to the neck-shaft angle of the hip joint.


Causes of blood methylomic variation for middle-aged women measured by the HumanMethylation450 array.
To address the limitations in current classic twin/family research on the genetic and/or environmental causes of human methylomic variation, we measured blood DNA methylation for 479 women (mean age 56 years) including 66 monozygotic (MZ), 66 dizygotic (DZ) twin pairs and 215 sisters of twins, and 11 random technical duplicates using the HumanMethylation450 array. For each methylation site, we estimated the correlation for pairs of duplicates, MZ twins, DZ twins, and siblings, fitted variance component models by assuming the variation is explained by genetic factors, by shared and individual environmental factors, and by independent measurement error, and assessed the best fitting model. We found that the average (standard deviation) correlations for duplicate, MZ, DZ, and sibling pairs were 0.10 (0.35), 0.07 (0.21), -0.01 (0.14) and -0.04 (0.07). At the genome-wide significance level of 10-7, 93.3% of sites had no familial correlation, and 5.6%, 0.1%, and 0.2% of sites were correlated for MZ, DZ, and sibling pairs. For 86.4%, 6.9%, and 7.1% of sites, the best fitting model included measurement error only, a genetic component, and at least one environmental component. For the 13.6% of sites influenced by genetic and/or environmental factors, the average proportion of variance explained by environmental factors was greater than that explained by genetic factors (0.41 vs. 0.37, P value <10-15). Our results are consistent with, for middle-aged woman, blood methylomic variation measured by the HumanMethylation450 array being largely explained by measurement error, and more influenced by environmental factors than by genetic factors.


[Levels of dissatisfaction of external users of the Instituto Nacional de Salud del Niño (National Institute of Child Health) in Lima, Peru].
To determine the changes in the levels of dissatisfaction of the external users of the Instituto Nacional de Salud del Niño, in Lima, Peru. Observational study based on a software to evaluate surveys completed by health care users (Software de Evaluación de Encuestas de Usuarios en Salud - SEEUS). The population consisted of every person accompanying external users to outpatient visits, inpatient visits or emergency treatment in the years 2008, 2009 and 2010. To calculate the sample size, last year's proportion of users dissatisfied with each service was used, with an estimated error of 5% at a 95% confidence level. A sample of 2051 people was obtained, which revealed a decreasing trend in the levels of dissatisfaction with outpatient visits, and an increased dissatisfaction in the medical capabilities dimension for hospitalization and emergency services. In addition, a dissatisfaction index lower than 0.20 was registered in the medical capabilities dimension for outpatient visits. In contrast, the highest dissatisfaction indexes were registered in the hygiene dimension for 2009 (0.25) and 2010 (0.25), and in the others dimension for emergency services in 2009 (0.25). The dimensions showing an increase in the dissatisfaction index were tangible aspects, hygiene and others for hospitalization and emergency services. On the other hand, the medical capabilities dimension for outpatient visits showed a decrease in the level of dissatisfaction over the evaluated period.


Gold nanoparticles promote osteogenic differentiation in human adipose-derived mesenchymal stem cells through the Wnt/β-catenin signaling pathway.
Gold nanoparticles (AuNPs) are attractive materials for use in biomedicine due to their physical properties. Increasing evidence suggests that several nanoparticles induce the differentiation of human mesenchymal stem cells into osteoblasts and adipocytes. In this study, we hypothesized that chitosan-conjugated AuNPs promote the osteogenic differentiation of human adipose-derived mesenchymal stem cells. For the evaluation of osteogenic differentiation, alizarin red staining, an alamarBlue(®) assay, and a quantitative real-time polymerase chain reaction analysis were performed. In order to examine specific signaling pathways, immunofluorescence and a western blotting assay were performed. Our results demonstrate that chitosan-conjugated AuNPs increase the deposition of calcium content and the expression of marker genes related to osteogenic differentiation in human adipose-derived mesenchymal stem cells at nontoxic concentrations. These results indicate that chitosan-conjugated AuNPs promote osteogenesis through the Wnt/β-catenin signaling pathway. Therefore, chitosan-conjugated AuNPs can be used as a reagent for promoting bone formation.


Congenital toxoplasmosis in England, Wales, and Northern Ireland: some epidemiological problems.
It has been suggested that congenital toxoplasmosis could be prevented by antenatal serological screening, followed by treatment or by termination of pregnancy if infection occurs. The only study of the incidence of congenital toxoplasmosis in England, Wales, and Northern Ireland took place more than 10 years ago. To obtain more recent figures laboratory reports of cases occurring from 1975 to 1980 were analysed. A total of 91 cases were reported over the six years. By criteria established to classify these infections only 34 were congenital, 20 were acquired postnatally, and 37 were unclassifiable. The mean annual number of cases of congenital toxoplasmosis was considerably smaller than that found in other recent studies. The condition could be underdiagnosed or rates of placental transmission could be lower in Britain than in other countries. Variation in reporting criteria of the laboratories made the data difficult to interpret. Improved diagnosis of congenital toxoplasmosis would not only clarify the epidemiology but would also help clinicians in management of suspected cases. Further antenatal surveys are necessary to assess the role of screening in the prevention of congenital toxoplasmosis.


A pragmatic checklist to identify pediatric ICU patients at risk for cardiac arrest or code bell activation.
In-hospital cardiac arrest is a rare event associated with significant morbidity and mortality. The ability to identify the ICU patients at risk for cardiac arrest could allow the clinical team to prepare staff and equipment in anticipation. This pilot study was completed at a large tertiary care pediatric intensive care unit to determine the feasibility of a simple checklist of clinical variables to predict deterioration. The daily checklist assessed patient risk for critical deterioration defined as cardiac arrest or code bell activation within 24h of the checklist screen. The Phase I checklist was developed by expert consensus and evaluated to determine standard diagnostic test performance. A modified Phase II checklist was developed to prospectively test the feasibility and bedside provider "number needed to train". For identifying patients requiring code bell activation, both checklists demonstrated a sensitivity of 100% with specificity of 76.0% during Phase I and 97.7% during Phase II. The positive likelihood ratio improved from 4.2 to 43.7. For identifying patients that had a cardiac arrest within 24h, the Phase I and II checklists demonstrated a sensitivity of 100% with specificity again improving from 75.7% to 97.6%. There was an improved positive likelihood ratio from 4.1 in Phase I to 41.9 in Phase II, with improvement of "number needed to train" from 149 to 7.4 providers. A novel high-risk clinical indicators checklist is feasible and provides timely and accurate identification of the ICU patients at risk for cardiac arrest or code bell activation.


Global exponential stability of recurrent neural networks for synthesizing linear feedback control systems via pole assignment.
Global exponential stability is the most desirable stability property of recurrent neural networks. The paper presents new results for recurrent neural networks applied to online computation of feedback gains of linear time-invariant multivariable systems via pole assignment. The theoretical analysis focuses on the global exponential stability, convergence rates, and selection of design parameters. The theoretical results are further substantiated by simulation results conducted for synthesizing linear feedback control systems with different specifications and design requirements.


Self-injurious behaviour: reviewing evidence for best practice.
This article brings together some of the research literature from the last 15 years concerning self-injurious behaviour (SIB) in people with learning disabilities. There is a focus on medical, biological and psychological research literature because of the important contribution these disciplines have made to the understanding of such behaviours. The article also explores contemporary evidence for the management of people with learning disabilities who present with SIB. The care and management of this group of people presents learning disability nurses with distinctively distressing challenges. Within the article definition, prevalence, topography, causation and evidence for best practice are outlined. The article concludes with a brief exploration of the role of the learning disability nurse in contributing to the health and wellbeing of this group of people.


Omega-3 polyunsaturated fatty acids: a potential new treatment of immune renal disease.
Omega-3 polyunsaturated fatty acids are among new treatments being tested for efficacy in immune renal disease. The principal omega-3 polyunsaturated fatty acids are eicosapentaenoic acid and docosahexaenoic acid. They are derived from alpha-linolenic acid, which is found mainly in marine lipids. Eicosapentaenoic acid and docosahexaenoic acid undergo biologic transformation into trienoic eicosanoids that alter inflammatory mediators and vascular reactivity, both of which are important in the pathogenesis of certain glomerular immune diseases. Investigators have shown that proteinuria was prevented and survival was prolonged in autoimmune models of nephritis after dietary supplementation with fish oil. Furthermore, vascular damage may be modified by the influence of eicosapentaenoic acid and docosahexaenoic acid on blood rheology, aggregation of platelets, and plasma lipids. In short-term clinical studies, omega-3 polyunsaturated fatty acids seem to diminish cyclosporine-induced nephrotoxicity and the attendant complication of hypertension, to inhibit inflammatory and atherogenic mechanisms in lupus nephritis, and to preserve renal function and reduce proteinuria in IgA nephropathy. Long-term clinical trials for testing fish oil in these three clinical conditions are under way to confirm or refute these apparent beneficial therapeutic results.


Lentivirus infection of macrophages.
The ovine and caprine lentiviruses infect monocytes, and the viral DNA is integrated into the cellular DNA. The provirus remains silent until the monocyte matures into a macrophage. Intrinsic to this maturation is the induction of a class of immediate early genes in the monocyte that includes the transcription factors JUN and FOS. These transcription factors are thought to couple short-term signals in the cell to long-term cellular differentiation by regulation of specific cellular genes. Thus, JUN and FOS bind to the AP-1 site in the promoters of cellular genes and activate their transcription, resulting in maturation of the monocyte into a macrophage. In addition, these cellular factors activate the same AP-1 sequence in the visna virus LTR, leading to transcriptional activation, full viral gene expression, and production of progeny virus. The expression of viral antigens in the context of MHC class II on the macrophage leads to the production of cytokines and a lymphoproliferative response that causes the lesions in specific target organs in an infected animal. We still understand only the framework of these events. The specific mechanisms by which viral genes alter macrophage gene expression and the molecular basis of different viral tropism for specific tissue macrophages, i.e. microglia, remain to be determined.


Epidermal hyperplasia and expansion of the interfollicular stem cell compartment in mutant mice with a C-terminal truncation of Patched1.
Hedgehog (Hh) signaling is conserved from flies to humans and is indispensable in embryogenesis and adulthood. Patched (Ptc) encodes a receptor for Hh ligands and functions as a tumor suppressor. PTCH1 mutations in humans are found in basal cell carcinoma (BCC) and irradiated Ptc1(+/-) mice recapitulate this phenotype. However, due to embryonic lethality associated with the Ptc1 null mutation, its normal function in embryonic and adult skin remains unknown. Here we describe the epidermal phenotypes of a spontaneous and viable allele of Ptc1, Ptc1(mes), in which the C-terminal domain (CTD) is truncated. Ptc1(mes/mes) embryos display normal epidermal and hair follicle development. Postnatal Ptc1(mes/mes) skin displays severe basal cell layer hyperplasia and increased proliferation, while stratification of the suprabasal layers is mostly normal. Interestingly, truncation of the Ptc1 CTD did not result in skin tumors. However, long term labeling studies revealed a greater than three-fold increase in label-retaining cells in the interfollicular epidermis of Ptc1(mes/mes) adults, indicating possible expansion of the epidermal stem cell compartment. Increased expression of regulators of epidermal homeostasis, c-Myc and p63, was also observed in Ptc1(mes/mes) adult skin. These results suggest that the CTD of Ptc1 is involved in regulating epidermal homeostasis in mature skin.


Variation in migration pattern, broodstock origin, and family productivity of coho salmon hatchery populations in British Columbia, Canada, derived from parentage-based tagging.
In salmonid parentage-based tagging (PBT) applications, entire hatchery broodstocks are genotyped, and subsequently, progeny can be nonlethally sampled and assigned back to their parents using parentage analysis, thus identifying their hatchery of origin and brood year (i.e., age). Inter- and intrapopulation variability in migration patterns, life history traits, and fishery contributions can be determined from PBT analysis of samples derived from both fisheries and escapements (portion of a salmon population that does not get caught in fisheries and returns to its natal river to spawn). In the current study of southern British Columbia coho salmon (Oncorhynchus kisutch) populations, PBT analysis provided novel information on intrapopulation heterogeneity among males in the total number of progeny identified in fisheries and escapements, the proportion of progeny sampled from fisheries versus escapement, the proportion of two-year-old progeny (jacks) produced, and the within-season return time of progeny. Fishery recoveries of coho salmon revealed heterogeneity in migration patterns among and within populations, with recoveries from north and central coast fisheries distinguishing "northern migrating" from "resident" populations. In northern migrating populations, the mean distance between fishery captures of sibs (brothers and sisters) was significantly less than the mean distance between nonsibs, indicating the possible presence of intrapopulation genetic heterogeneity for migration pattern. Variation among populations in productivity and within populations in fish catchability indicated that population selection and broodstock management can be implemented to optimize harvest benefits from hatcheries. Application of PBT provided valuable information for assessment and management of hatchery-origin coho salmon in British Columbia.


Upgrading small WWTPs in the autonomous province of Trento (Italy) by alternating oxic/anoxic process: a demonstration study.
The paper deals with the real application of a strategy, based on process control automation and remote on-line supervision, for the wastewater treatment in a piedmont. Three existing small wastewater treatment plants were selected to be upgraded and to be included into a network remotely supervised. Further, the potentialities of the process control automation were enhanced by the appropriate upgrading of the whole plant. A consolidated alternating nitrification and denitrification process was applied for the biological treatment. The selected plants well represent the area of Autonomous Province of Trento because they are characterized by remarkable seasonal fluctuation. After five months of experimentation data processed show the real stable high quality of the treated effluent in terms of total nitrogen content. Moreover, the power requirements are significantly reduced according the correct energy policy. The performances of the new biological process applied and the economical balance, put in evidence the gain by chosing the AC technology for upgrading small WWTPs.


Embryonic poly(A)-binding protein stimulates translation in germ cells.
The function of poly(A)-binding protein 1 (PABP1) in poly(A)-mediated translation has been extensively characterized. Recently, Xenopus laevis oocytes and early embryos were shown to contain a novel poly(A)-binding protein, ePABP, which has not been described in other organisms. ePABP was identified as a protein that binds AU-rich sequences and prevents shortening of poly(A) tails. Here, we show that ePABP is also expressed in X. laevis testis, suggesting a more general role for ePABP in gametogenesis. We find that ePABP is conserved throughout vertebrates and that mouse and X. laevis cells have similar tissue-specific ePABP expression patterns. Furthermore, we directly assess the role of ePABP in translation. We show that ePABP is associated with polysomes and can activate the translation of reporter mRNAs in vivo. Despite its relative divergence from PABP1, we find that ePABP has similar functional domains and can bind to several PABP1 partners, suggesting that they may use similar mechanisms to activate translation. In addition, we find that PABP1 and ePABP can interact, suggesting that these proteins may be bound simultaneously to the same mRNA. Finally, we show that the activity of both PABP1 and ePABP increases during oocyte maturation, when many mRNAs undergo polyadenylation.


Carotid sinus reflex vasoconstriction in right coronary circulation of dog and pig.
This study was designed to evaluate the effects of the sympathetic nervous system on right ventricular and right coronary hemodynamics in the anesthetized vagotomized dog and pig during constant-pressure coronary perfusion. Carotid occlusion in the dog resulted in an increase in heart rate, aortic pressure, right ventricular dP/dt, and a sustained 13% decrease in right coronary artery blood flow and a 14% increase in coronary vascular resistance. This coronary vasoconstriction was not modified during beta-blockade with propranolol but was prevented by alpha- blockade with phentolamine. Similar hemodynamic data was obtained in the pig and, in addition, it was found that the O2 consumption of the right ventricle is low (4.8 +/- 0.4 ml O2.min-1.100 g-1) and is only slightly increased (5.l5 +/- 0.8 ml.min-1.100 g-1) during carotid occlusion. These data suggest that baroreflex-mediated sympathetic stimulation to the heart evokes only a minor increase in metabolic activity in the right ventricular myocardium so that the direct neural (alpha-vasoconstrictor) effect predominates over the metabolic vasodilator mechanism.


Spatial-temporal and cancer risk assessment of selected hazardous air pollutants in Seattle.
In the Seattle Air Toxics Monitoring Pilot Program, we measured 15 hazardous air pollutants (HAPs) at 6 sites for more than a year between 2000 and 2002. Spatial-temporal variations were evaluated with random-effects models and principal component analyses. The potential health risks were further estimated based on the monitored data, with the incorporation of the bootstrapping technique for the uncertainty analysis. It is found that the temporal variability was generally higher than the spatial variability for most air toxics. The highest temporal variability was observed for tetrachloroethylene (70% temporal vs. 34% spatial variability). Nevertheless, most air toxics still exhibited significant spatial variations, even after accounting for the temporal effects. These results suggest that it would require operating multiple air toxics monitoring sites over a significant period of time with proper monitoring frequency to better evaluate population exposure to HAPs. The median values of the estimated inhalation cancer risks ranged between 4.3 × 10⁻⁵ and 6.0 × 10⁻⁵, with the 5th and 95th percentile levels exceeding the 1 in a million level. VOCs as a whole contributed over 80% of the risk among the HAPs measured and arsenic contributed most substantially to the overall risk associated with metals.


Transport and metabolism of glucose in an insulin-secreting cell line, beta TC-1.
Kinetic characteristics of glucose transport and glucose phosphorylation were studied in the islet cell line beta TC-1 to explore the roles of these processes in determining the dependence of glucose metabolism and insulin secretion on external glucose. The predominant glucose transporter present was the rat brain/erythrocyte type (Glut1), as determined by RNA and immunoblot analysis. The liver/islet glucose transporter (Glut2) RNA was not detected. The functional parameters of zero-trans glucose entry were Km = 9.5 +/- 2 mM and Vmax = 15.2 +/- 2 nmol min-1 (microL of cell water)-1. Phosphorylation kinetics of two hexokinase activities were characterized in situ. A low-Km (0.036 mM) hexokinase with a Vmax of 0.40 nmol min-1 (microL of cell water)-1 was present along with a high-Km (10 mM) hexokinase, which appeared to conform to a cooperative model with a Hill coefficient of about 1.4 and a Vmax of 0.3 nmol min-1 (microL of cell water)-1. Intracellular glucose at steady state was about 80% of the extracellular glucose from 3 to 15 mM, and transport did not limit metabolism in this range. In this static (nonperifusion) system, 2-3 times more immunoreactive insulin was secreted into the medium at 15 mM glucose than at 3 mM. The dependence of insulin secretion on external glucose roughly paralleled the dependence of glucose metabolism on external glucose. Simulations with a model demonstrated the degree to which changes in transport activity would affect intracellular glucose levels and the rate of the high-Km hexokinase (with the potential to affect insulin release).


Parents' reflective functioning and their agreement on treatment goals in attachment-based family therapy for sexual and gender minority young adults and their nonaccepting parents.
This study examined the trajectories of parents' reflective functioning over the course of individual preparatory sessions with the therapist in attachment-based family therapy (ABFT) for Israeli sexual minority young adults and their nonaccepting parents, and whether such reflective functioning was associated with parents' agreement on the goals for subsequent conjoint corrective attachment sessions. Three parents with good agreement on treatment goals were compared to three parents with poor agreement on treatment goals. Parents' in-session reflectivity regarding their child's experience of nonacceptance, and regarding the quality of their relationship with their child, was observationally measured at the session level. Reflective functioning increased over time among those parents who reported good agreement with the treatment goals. There were no changes in parents' reflective functioning in the poor agreement group. Findings suggest the potential importance of facilitating parents' reflective functioning in ABFT, before conducting conjoint corrective attachment sessions. Research and clinical implications are discussed.


Dopamine, urges to smoke, and the relative salience of drug versus non-drug reward.
When addicted individuals are exposed to drug-related stimuli, dopamine release is thought to mediate incentive salience attribution, increasing attentional bias, craving and drug seeking. It is unclear whether dopamine acts specifically on drug cues versus other rewards, and if these effects correspond with craving and other forms of cognitive bias. Here, we administered the dopamine D2/D3 agonist pramipexole (0.5 mg) to 16 tobacco smokers in a double-blind placebo-controlled crossover design. Visual fixations on smoking and money images were recorded alongside smoking urges and fluency tasks. Pramipexole attenuated a marked bias in initial orienting towards smoking relative to money but did not alter a maintained attentional bias towards smoking. Pramipexole decreased urges to smoke retrospectively after the task but not on a state scale. Fewer smoking words were generated after pramipexole but phonological and semantic fluency were preserved. Although these treatment effects did not correlate with each other, changes in initial orienting towards smoking and money were inversely related to baseline scores. In conclusion, pramipexole can reduce the salience of an addictive drug compared with other rewards and elicit corresponding changes in smoking urges and cognitive bias. These reward-specific and baseline-dependent effects support an 'inverted-U' shaped profile of dopamine in addiction.


[The binding ability of insulin-related peptides as the clue to their functional role in phylogenesis. Introduction to the problem].
The common plan of structure of the main peptides of the vertebrate insulin family--insulin itself, IGF-I, IGF-II, and relaxin--has distinct structural features. Each of the peptides performs its characteristic function. However, overlapping of insulin and IGF-I actions and its stability in the vertebrate phylogenesis have formed the concept of their regulation of growth and metabolism as a function fixed in phylogenesis for a certain type of structures. At the same time, study of insulin-related peptides in invertebrates has revealed the wider spectrum, than in vertebrates, of biological effects; this indicated that the similarity of the total structure designs not sufficient for judging about their functional role. Functional possibilities of a regulatory peptide depend fundamentally on its capability for binding to the receptor realizing its biological action. However, the binding ability has a wider significance than merely transmission of biological signals. Thus, IGF-II when interacting with receptors realizing its biological effects, also binds to he IGF-2 receptor limiting its action and, besides, to the binding proteins (BP) modulating its action. The entire cycle of interactions occurs in the organism at different affinity levels. Meanwhile insulin interacts neither with IGF-2 receptor nor with BP. In this case, specificity and sequence of interaction with each of receptors or with protein are due not to the general design of the peptide structure, but rather to structure of individual submolecular determinants--binding domains. The leading role in disclosure of composition and structure of these domains is played by the "mutant-ligand" approach evaluating affinity of modified analogs. To analyze role of structural elements of the binding domains, the author proposes the system of estimation of affinity of the studied analogs. The present work, alongside with consideration of methodical aspects of the forthcoming analysis, is an introduction to the problem of organization of the binding domains related directly with functional role of peptides of the insulin type. The proposed analysis is due to the necessity of specification of this organization both within one molecule and in different molecules with a similar plan of structure on the basis of nor always unambiguous literature data and of clarification of principles of structure of these domains.


Cross-sectional study of hepatitis B virus infection in rural communities, Central African Republic.
As most data on hepatitis in resource-poor countries relate to urban communities, surveys in the rural environment are necessary to determine the 'true' prevalence of these viral infections. We undertook a survey to determine the prevalence of hepatitis B virus (HBV) infection in an apparently healthy rural population in the Central African Republic (CAR). The cross-sectional study was based on dried blood spots (DBS) from 273 people recruited in four prefectures (Lobaye, Nana-Mambéré, Ouham and Ouaka). Eluates from DBS were tested with commercial ELISA kits to detect markers of HBV infection. DBS were directly used for DNA extraction, followed by PCR and genotyping based on preS/S gene sequences. The overall prevalence of HBc antibodies was 27.1% (Lobaye 29%, Nana-Mambéré 28%, Ouaka 29% and Ouham 23%) and that of HBsAg was 10.6% (Lobaye 9%, Nana-Mambéré 9%, Ouaka 19% and Ouham 8%), with no statistically significant difference among the surveyed communities. Nineteen sequences obtained from 74 anti-HBc-positive patients all belonged to genotype E. Risk factor analysis of HBV infection pointed to sexual transmission of the virus. The prevalence of HBV is high in rural communities in the CAR and comparable to that observed in urban areas. In addition, genotype E is prevalent in these areas. These findings underline the importance of instituting a programme of active HBV surveillance and vaccination of the population.


A Review on Theragnostic Applications of Micrornas and Long Non- Coding RNAs in Colorectal Cancer.
Colorectal cancer (CRC) is a heterogeneous malignancy leading to increased mortality and poor prognosis due to the lack of efficient early diagnostics. Metastasis of the tumor being the most common cause of mortality is accountable for almost 90% of CRC associated deaths. Intensified screening procedures and molecular target identification has inflated the median survival rate of in CRC patients. microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have come forward as potential targets for developing a novel approach in CRC theragnostics. Non-coding RNA (ncRNAs) sequences are abundantly present and thereby play a vital role in several biological processes such as cellular organization, cell fate determination, proliferation, apoptosis, tissue homeostasis maintenance as well as pathological conditions such as cancer by acting as post transcriptional regulators of gene expression. Several studies have highlighted the involvement of these ncRNAs in CRC development. However, the molecular mechanism involved in regulating CRC has not been clearly elucidated. This review, throws light upon the several non-coding RNAs involved in CRC with a focus on novel mechanisms of action, recent advances in the regulatory mechanisms that control the gene expression related to carcinogenesis. Furthermore, the potential role of ncRNAs as diagnostic as well as therapeutic targets has been reviewed.


Robert May memorial lecture: Advances in reconstructive venous surgery.
During the last decade progress in reconstructive venous surgery is mainly based on improvements of preoperative diagnosis and on changed surgical techniques (e.g. intraoperative lumen control by vascular endoscopy or angiography; temporary arteriovenous fistulas as protective measure; external supported vascular prostheses as substitutes). The main indications include acute ilio-femoral phlebothrombosis, vein injuries and in selected cases the substitution of big veins in extended tumor surgery. The late results of venous thrombectomy in the ilio-femoral segment could be remarkably improved due to changed techniques of lumen restoration (combined ring- and balloon-disobliteration, temporary a.v. fistula in inguinal or popliteal position). In patients with persisting central occlusions in the iliac veins a second repair may be considered using a suprainguinal cross-over graft (external supported e-PTFE-prosthesis). For post-thrombotic syndrome with pronounced deep venous insufficiency reconstructive procedures afford a critical selection of patients (Palma-operation, Husni-May-operation, interposition of valve bearing venous segments in the superficial femoral vein). The remarkable contribution given by R. May to the advances in venous surgery is illustrated.


Clinical and pharmaceutical applications of affinity ligands in capillary electrophoresis: A review.
Affinity capillary electrophoresis (ACE) is a separation technique that combines a biologically-related binding agent with the separating power and efficiency of capillary electrophoresis. This review will examine several classes of binding agents that have been used in ACE and applications that have been described for the resulting methods in clinical or pharmaceutical analysis. Binding agents that will be considered are antibodies, aptamers, lectins, serum proteins, carbohydrates, and enzymes. This review will also describe the various formats in which each type of binding agent has been used in CE, including both homogeneous and heterogeneous methods. Specific areas of applications that will be considered are CE-based immunoassays, glycoprotein/glycan separations, chiral separations, and biointeraction studies. The general principles and formats of ACE for each of these applications will be examined, along with the potential advantages or limitations of these methods.


Klotho as a potential biomarker and therapy for acute kidney injury.
Klotho is a single-pass transmembrane protein that is highly expressed in the kidney and is known to act as a coreceptor for fibroblast growth factor 23. The extracellular domain can be produced independently or shed from membrane-bound Klotho and functions as an endocrine substance with multiple functions including antioxidation, modulation of ion transport, suppression of fibrosis, and preservation of stem cells. Emerging evidence has revealed that Klotho deficiency is an early event in acute kidney injury (AKI), and a pathogenic factor that exacerbates acute kidney damage and contributes to long-term consequences. Restoration by exogenous supplementation or stimulation of endogenous Klotho might prevent and ameliorate injury, promote recovery, and suppress fibrosis to mitigate development of chronic kidney disease. Although data are still emerging, in this Perspectives article we discuss why this renal-derived protein is a highly promising candidate as both an early biomarker and therapeutic agent for AKI.


Phytochemical-derived tumor-associated macrophage remodeling strategy using Phoenix dactylifera L. boosted photodynamic therapy in melanoma via H19/iNOS/PD-L1 axis.
The tumor microenvironment (TME) represents a barrier to PDT efficacy among melanoma patients. The aim of this study is to employ a novel muti-tactic TME-remodeling strategy via repolarization of tumor-associated macrophages (TAMs), the main TME immune cells in melanoma, from the pro-tumor M2 into the antitumor M1 phenotype using Phoenix dactylifera L. (date palm) in combination with PDT. Screening of different date cultivars was employed to choose extracts of selective toxicity to melanoma and TAMs, not normal macrophages. Potential extracts were then fractionated and characterized by gas chromatography-mass spectrometry (GC-MS). Finally, the efficacy and the potential molecular mechanism of the co-treatment were portrayed via quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Initial screening resulted in the selection of the two Phoenix dactylifera L. cultivars Safawi and Sukkari methanolic extracts. Sukkari showed superior capacity to revert TAM phenotype into M1 as well as more prominent upregulation of M1 markers and repression of melanoma immunosuppressive markers relative to positive control (resiquimod). Molecularly, it was shown that PDT of melanoma cells in the presence of the secretome of repolarized TAMs surpassed the monotherapy via the modulation of the H19/iNOS/PD-L1immune-regulatory axis. This study highlights the potential utilization of nutraceuticals in combination with PDT in the treatment of melanoma to provide a dual activity through alleviating the immune suppressive TME and potentiating the anti-tumor responses.


Effects of antioxidant treatment on seminal parameters in patients undergoing in vitro fertilization.
The aim of this non controlled trial was to assess whether a therapy with an antioxidant supplement may improve spermatozoa quality in terms of number, motility, morphology and a higher number of successful conceptions in patients with oligoasthenoteratozoospermia undergoing cycles of medically assisted reproduction by intracytoplasmic sperm injection (ICSI). 32 patients registered at A.G.I. Medica (Siena) medically assisted reproduction centre affected by fertility problems associated with oligoasthenoteratozoospermia were included in the study. Semen analysis were evaluated according to World Health Organization 2010, before and after treatment. Moreover, we used colorimetric tests to assess oxidative stress. After evaluating oocyte fertilisation rate and the quality of embryos obtained, data were statistically analysed. Microscopy examination after the therapy, showed a general improvement in sperm parameters (number of sperms, progressive motility, viability and normal morphology) in both baseline and capacitated; also the levels of oxidative stress was notably lower after the treatment. Morever we evaluated the outcome of the IVF treatment, the percentage of fertilization and the number of embryos obtained, all the parameters was significantly higher in the N1 group. The outcomes of this trial seem to suggest that the administration of our food supplement improve semen parameters and that the evaluation of oxidative stress levels may become a diagnostic tool to assess male infertility in patients undergoing ART cycle.