Allele frequency divergence reveals ubiquitous influence of positive selection in Drosophila.
Resolving the role of natural selection is a basic objective of evolutionary biology. It is generally difficult to detect the influence of selection because ubiquitous non-selective stochastic change in allele frequencies (genetic drift) degrades evidence of selection. As a result, selection scans typically only identify genomic regions that have undergone episodes of intense selection. Yet it seems likely such episodes are the exception; the norm is more likely to involve subtle, concurrent selective changes at a large number of loci. We develop a new theoretical approach that uncovers a previously undocumented genome-wide signature of selection in the collective divergence of allele frequencies over time. Applying our approach to temporally resolved allele frequency measurements from laboratory and wild Drosophila populations, we quantify the selective contribution to allele frequency divergence and find that selection has substantial effects on much of the genome. We further quantify the magnitude of the total selection coefficient (a measure of the combined effects of direct and linked selection) at a typical polymorphic locus, and find this to be large (of order 1%) even though most mutations are not directly under selection. We find that selective allele frequency divergence is substantially elevated at intermediate allele frequencies, which we argue is most parsimoniously explained by positive-not negative-selection. Thus, in these populations most mutations are far from evolving neutrally in the short term (tens of generations), including mutations with neutral fitness effects, and the result cannot be explained simply as an ongoing purging of deleterious mutations.


An integrative approach to identify YB-1-interacting proteins required for cisplatin resistance in MCF7 and MDA-MB-231 breast cancer cells.
The Y-box binding protein 1 (YB-1) is a multifunctional protein that affects transcription, splicing, and translation. Overexpression of YB-1 in breast cancers causes cisplatin resistance. The exact mechanism by which YB-1 confers cisplatin resistance is unknown. The aim of the present study was to identify, using mass spectrometry, proteins that interact with YB-1 that are important for cisplatin resistance in two breast cancer cell lines, namely MCF7 and MDA-MB-231. A tagged YB-1 construct was used to identify proteins interacting directly with YB-1 in breast cancer cells. We then focused on proteins that are potentially involved in breast cancer progression based on the ONCOMINE public microarray database. Genes encoding for these YB-1-interacting proteins were examined in the public NCBI comparative genomic hybridization database to determine whether they are localized to regions of chromosomes that are rearranged in breast cancer tissues. From these analyses, we generated a list of proteins potentially involved in cisplatin resistance. Cisplatin dose-response curves were constructed in MCF7 and MDA-MB-231 transfected with four siRNA corresponding to each of these YB-1 interactors to identify proteins significantly affecting cisplatin sensitivity upon gene silencing. Depletion of only the X-linked ribosomal protein S4 (RPS4X) resulted in consistent resistance to cisplatin in both cell lines with at least three different siRNA sequences against RPS4X. Further analyses indicated that the knock down of RPS4X decreased DNA synthesis, induced cisplatin resistance, and is equivalent to the overexpression of YB-1 in both MCF7 and MDA-MB-231 cells. These results suggest that the RPS4X/YB-1 complex is a significant potential target to counteract cisplatin resistance in breast cancer.


Effect of different levels of gibberellic acid and kinetin on quality and self-life of banana (Musa spp.) fruits.
Artificial fruit ripening agents have become increasingly popular in recent years. During the off-seasons, various ripening agents are used for the ripening of banana fruits. The effects of various ripening agents of banana fruits (variety: Malbhog) were studied. Seven treatments consisting of distilled water, Gibberellic acid (GA3) @ 100 ppm, GA3 @ 200 ppm, GA3 @ 300 ppm, kinetin @ 3 ppm, kinetin @ 5 ppm, and kinetin @ 7 ppm were laid out in a Completely Randomized Design with three replications. The maximum loss in weight (9.195%), Total Soluble Solids (20.33˚B), the highest color score (6), pH (4.767), Total Soluble Solids/Titratable Acidity TA (34.23), and Pulp peel ratio (2.84) were observed in banana sprayed with distilled water whereas the minimum value for TSS (12.67˚B), pulp peel ratio (1.813), peel color rating (2.67), TSS/TA (15.32) were observed in GA3 @ 300 ppm treated fruits. The maximum (32.67 days) and the minimum (18.33 days) shelf-life was observed in GA3 @ 300 ppm and distilled water respectively. This research will help in the regulation of ripening as per the needs of consumers and the distance of the market. In horticulture, it will help to extend storage life and reduce quality and economic loss.


[Reliability and validity of the standardized Mini Mental State Examination in the diagnosis of mild dementia in Turkish population].
Reliability and validity of the Mini Mental State Examination in differentiating mild dementia from normal controls in Turkish population. The Standardized Mini Mental State Examination (SMMSE) and its instruction were translated into Turkish. A total of 212 subjects with mean age of 77 +/- 6, were recruited for the study. 71 were diagnosed to be demented and 141 were evaluated as normal controls. The scale total score was analysed for discriminant validity using Student's t-test. Sensitivity, specificity, positive and negative predictive values and kappa score were calculated for all of the scores between 18 and 29. Kappa value was calculated for the comparison of the dementia diagnosis between the two investigators using the best cut off score obtained in the analysis above. Statistical analysis revealed that the Turkish version of the SMMSE has high discriminant validity and interrater reliability in the diagnosis of mild dementia. The cut off score 23/24 was found to have the highest sensitivity (0.91), specificity (0.95), positive and negative predictive values (0.90 and 0.95) and kappa score (0.86). Interrater reliability analysis showed high correlation (r:0.99) and kappa value (0.92). The results of this study showed that the Turkish version of the SMMSE has high reliability and validity for the diagnosis of mild dementia in Turkish population.


Cellulose nanocrystals-based bio-composite optical materials for reversible colorimetric responsive films and coatings.
Photonic materials with a tunable chiral nematic structure that can selectively reflect light dynamically are valuable for applications in smart responsive materials. Here, we prepared potential photonic composites with a chiral nematic structure by forming cellulose nanocrystals (CNCs) and waterborne polyurethane (WPU) composites with different compositions on different substrates by evaporation-induced self-assembly. With increasing WPU content, the reflected wavelength increased from 400 to 680 nm, which was mainly caused by the increase of the chiral nematic pitch. In addition, the mechanical properties were better for higher WPU content. WPU was sensitive to small amounts of moisture in ethanol owing to the swollen WPU after absorbing water will increase the helical pitch. The reversible red shift induced by moisture was approximately 100 nm. When wood was used as the substrate, the CNCs still self-assembled to form chiral nematic structures and the adhesion forces of the composites to the wood substrate were strong. By using MgCl2 solution as an ink, invisible patterns can be written on the coating, which can be revealed temporarily by ethanol. In addition, the invisible pattern of photonic coating is rewritable. The easily prepared environmentally friendly photonic composite has great potential in sensors, anti-counterfeiting labels and smart decorative coatings.


Intragastric and atomized administration of canagliflozin inhibit inflammatory cytokine storm in lipopolysaccharide-treated sepsis in mice: A potential COVID-19 treatment.
To date, drugs to attenuate cytokine storm in severe cases of Corona Virus Disease 2019 (COVID-19) are not available. In this study, we investigated the effects of intragastric and atomized administration of canagliflozin (CAN) on cytokine storm in lung tissues of lipopolysaccharides (LPS)-induced mice. Results showed that intragastric administration of CAN significantly and widely inhibited the production of inflammatory cytokines in lung tissues of LPS-induced sepsis mice. Simultaneously, intragastric administration of CAN significantly improved inflammatory pathological changes of lung tissues. Atomized administration of CAN also exhibited similar effects in LPS-induced sepsis mice. Furthermore, CAN significantly inhibited hypoxia inducible factor 1α (HIF-1α) and phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) protein levels in LPS-treated lung tissues. These results indicated that CAN might attenuate cytokine storm and reduce the inflammatory symptoms in critical cases in COVID-19. Its action mechanism might involve the regulation of HIF-1α and glycolysis in vivo. However, further studies about clinical application and mechanism analysis should be validated in the future.


1-Tosyl-2-[(1-tosyl-1H-benzimidazol-2-yl)methyl-sulfanyl]-1H-benzimidazole.
In the title compound, C(29)H(24)N(4)O(4)S(3), the two N-tosyl-benzimidazolyl unit are connected through a -S-CH(2)- fragment, the dihedral angle between the benzimidazole rings being 76.09 (5)°. The methyl-thio group is disordered with respect to exchange of the S and C atoms in a 0.547 (4):0.453 (4) ratio. In the crystal, C-H⋯O and C-H⋯π inter-actions connect adjacent mol-ecules into infinite layers parallel to the ab plane. The crystal packing is further stabilized by a π-π inter-action [centroid-centroid separation = 3.5187 (4) Å].


Effect of fluoride on uptake of D-glucose by isolated epithelial cells of rat intestine.
Inhibition on uptake of D-glucose by isolated intestinal epithelial cells (IIEC) was observed when the fluoride concentration ranged between 0.25 and 5 mM. Active transport was almost completely inhibited at 5 mM. When CaCl2 was added to fluoride, the inhibitory effect on glucose uptake was abolished. Preincubation of IIEC with different concentrations of fluoride (2.5-5.0 mM) for different intervals of time (2-20 min) at different pH levels (6.2-7.8) and temperatures (0-37 degrees C) revealed that the conditions which led to higher uptake of fluoride by IIEC produced maximum inhibition. The degree of inhibition was not appreciably altered by a change in glucose concentrations. A concentration-dependent effect of fluoride on lactic acid and carbon dioxide production by IIEC was also observed.


Hypotension in Nursing Home Residents on Antihypertensive Treatment: Is it Associated with Mortality?
To assess whether low systolic blood pressure (SBP) or diastolic blood pressure (DBP) due to antihypertensive medications might be related to mortality among nursing home (NH) residents. Observational, longitudinal. Nursing home. Age ≥60 years, receiving antihypertensive medications. Demographic characteristics, mobility status, number of chronic diseases and drugs, nutritional status, and antihypertensive medications were noted. At the first visit, we recorded blood pressure (BP) measurements of last 1 year, which were measured regularly at 2-week intervals and considered their mean values. SBP and DBP thresholds were analyzed for mortality by ROC analysis. Multivariate Cox regression analyses were performed to determine factors related to mortality. The sample included 253 residents with a mean age of 75.7 ± 8.7 years, and 66% were male. Residents were evaluated at a mean follow-up time of 14.3 ± 5.2 months (median: 15) for short-term mortality and 31.6 ± 14.3 months (median: 40) for long-term mortality. The prevalence of low SBP (≤110 mm Hg) and low DBP (≤65 mm Hg) was 34.8% and 15.8%, respectively. In follow-up, the short-term mortality rate was 21.7% (n = 55) and the long-term mortality rate was 42.2% (n = 107). Low SBP (≤110 mm Hg) was related to mortality in short- and long-term follow-ups [short-term follow-up: hazard ratio (HR) 3.7, 95% confidence interval (CI) 1.5-8.6, P = .01; long-term follow-up: HR 1.8, 95% CI 1.1-3.0, P = .02], adjusted for age, mobility status, nutritional state, and total number of diseases and drugs. Low DBP (≤65 mm Hg) was related to mortality in short- and long-term follow-ups [short-term follow-up: HR 3.0, 95% CI 1.2-7.8, P = .02, long-term follow-up: HR 2.8, 95% CI 1.5-5.2, P = .001], adjusted for age, mobility status, nutritional state, and total number of diseases and drugs. Systolic hypotension was found in more than one-third of the NH residents receiving antihypertensive treatment. Low SBP and DBP were significant factors associated with mortality. Particular attention should be paid to prevent low SBP and DBP in NH residents on antihypertensive treatment.


Reduced activity of vertically acting motoneurons during convergence.
Previous studies have revealed unexpected relationships between the firing rates of horizontally acting motoneurons and vergence. During a vergence task, for example, antidromically identified abducens internuclear neurons show a negative correlation between vergence angle and firing rate, which is the opposite of the modulation displayed by the medial rectus motoneurons to which they project. For a given horizontal eye position, medial rectus motoneurons discharge at a higher rate if the eyes are converged than if the same eye position is reached during a task that requires version; paradoxically, however, the horizontal rectus eye muscles show corelaxation during vergence. These complex and unexpected relationships inspired the present author to investigate whether the tonic firing rates of vertically acting motoneurons in oculomotor nucleus are correlated with vergence angle. Monkeys were trained to fixate a single, randomly selected, visual target among an array of 60 red plus-shaped LEDs, arranged at 12 different distances in three-dimensional space. The targets were arranged to permit dissociation of vertical eye position and vergence angle. Here I report, for the first time, that most vertically acting motoneurons in oculomotor nucleus show a significant negative correlation between tonic firing rate and vergence angle. This suggests the possibility that there may be a general corelaxation of extraocular muscles during vergence.NEW & NOTEWORTHY An array of 60 plus-shaped LEDs, positioned at various locations in three-dimensional space, was used to elicit conjugate and disjunctive saccades while single neurons in oculomotor nucleus were recorded from rhesus monkeys. This study demonstrates that most vertically acting motoneurons in oculomotor nucleus discharge at a lower rate when the eyes are converged.


Cross-Sectional Comparative Study on Central Sensitization-Psychosocial Associated Comorbidities and Psychological Characteristics in Breast Cancer Survivors with Nociceptive Pain and Pain with Neuropathic Features and without Pain.
The frequency of a high Central Sensitization Inventory (CSI) total score and the prevalence of pain have already been established among breast cancer survivors (BCS). However, the psychological factors' influence based on the clinical features of pain is still unknown, as well as BCS characteristics with no pain. Thus, our main aim was to evaluate the presence of a high CSI total score in BCS with pain and compare it with BCS without pain and to evaluate the influence of psychosocial factors. A cross-sectional comparative study was designed to compare BCS with nociceptive pain (n = 19), pain with neuropathic features (n = 19) or no pain (n = 19), classified by the Leeds Assessment of Neuropathic Symptoms and Signs (LANSS). CSI, pain catastrophizing, fear of movement, anxiety and depression symptoms were analyzed and compared among the three groups. The CSI total score was higher in both BCS pain groups compared to BCS without pain, but there were no statistical differences between the pain groups. The same observation was made when comparing pain catastrophizing. The neuropathic feature group showed greater levels of fear of movement, anxiety and depression compared to the no pain group. Thus, CS-psychosocial associated comorbidities and pain-catastrophizing thoughts were more prevalent among BCS with pain, regardless of the clinical features of pain. BCS with neuropathic pain features showed greater psychological disturbances.


A novel TAT-mitochondrial signal sequence fusion protein is processed, stays in mitochondria, and crosses the placenta.
Mutations in nuclear and mitochondrial genomes can lead to defects in mitochondrial function. To date, repair of these defects with exogenous proteins or gene transfer has been difficult with either viral or nonviral vectors. We hypothesized that TAT fusion proteins would cross both mitochondrial membranes and that incorporation of a mitochondrial signal sequence into a TAT fusion protein would allow processing and localization of exogenous proteins in mitochondria. A TAT-mitochondrial malate dehydrogenase signal sequence (mMDH)-enhanced green fluorescent protein (eGFP) fusion protein was constructed. TAT-mMDH-eGFP allowed rapid transduction and localization of fusion protein into mitochondria of multiple cell types. In contrast, TAT-GFP, without a mitochondrial signal sequence, rapidly transduced into cells and mitochondria, displayed pseudo-first-order kinetics, but did not remain there. Mice injected 5 days prior with TAT-mMDH-eGFP had detectable eGFP activity in multiple tissue types. Western blotting of cytosolic and mitochondrial fractions isolated from their livers confirmed eGFP localization to mitochondria and that the mMDH transit peptide was recognized and processed. Furthermore, TAT-mMDH-eGFP fusion protein injected into pregnant mice crossed the placenta and was detectable in both the fetus and the newborn pups. TAT fusion proteins containing a mitochondrial signal sequence are a viable method to localize proteins to mitochondria.


How do sex hormones modify arrhythmogenesis in long QT syndrome? Sex hormone effects on arrhythmogenic substrate and triggered activity.
Gender differences in cardiac repolarization and the arrhythmogenic risk of patients with inherited and acquired long QT syndromes are well appreciated clinically. Enhancing our knowledge of the mechanisms underlying these differences is critical to improve our therapeutic strategies for preventing sudden cardiac death in such patients. This review summarizes the effects of sex hormones on the expression and function of ion channels that control cardiac cell excitation and repolarization as well as key proteins that regulate Ca(2+) dynamics at the cellular level. Moreover, it examines the role of sex hormones in modifying the dynamic spatiotemporal (regional and transmural) heterogeneities in action potential duration (eg, the arrhythmogenic substrate) and the susceptibility to (sympathetic) triggered activity at the tissue, organ, and whole animal levels. Finally, it explores the implications of these effects on the management of patients with LQTS.


Gradients in the in vivo intestinal stem cell compartment and their in vitro recapitulation in mimetic platforms.
Intestinal tissue, and specifically its mucosal layer, is a complex and gradient-rich environment. Gradients of soluble factor (BMP, Noggin, Notch, Hedgehog, and Wnt), insoluble extracellular matrix proteins (laminins, collagens, fibronectin, and their cognate receptors), stromal stiffness, oxygenation, and sheer stress induced by luminal fluid flow at the crypt-villus axis controls and supports healthy intestinal tissue homeostasis. However, due to current technological challenges, very few of these features have so far been included in in vitro intestinal tissue mimetic platforms. In this review, the tightly defined and dynamic microenvironment of the intestinal tissue is presented in detail. Additionally, the authors introduce the current state-of-the-art intestinal tissue mimetic platforms, as well as the design drawbacks and challenges they face while attempting to capture the complexity of the intestinal tissue's physiology. Finally, the compositions of an "idealized" mimetic system is presented to guide future developmental efforts.


CT and pathologic correlation of Tc-99m medronate splenic uptake in metastatic breast carcinoma.
The CT and autopsy findings in a case of bone tracer uptake by a spleen infiltrated with metastatic breast carcinoma is presented. Calcium could not be demonstrated by either CT or special stains of histologic sections of the spleen. The mechanisms that have been postulated for radiotracer uptake by the spleen are reviewed in relationship to the correlation by this case. In addition, bone tracer uptake did not occur in five other sites of metastatic disease.


Facilitators and challenges to exclusive breastfeeding in Belagavi District, Karnataka, India.
A primary objective of this study was to identify specific facilitators and challenges around exclusive breastfeeding (EBF) in our community in India, from the perspective of breastfeeding mothers and their support networks. We conducted eight focus groups incorporating 75 women and their support networks in the Belagavi District, Karnataka State, India. We used a directed content analysis to guide the analysis. The specific facilitator that emerged as a theme, broad support for and knowledge about breastfeeding on the individual, family and community levels, was a seeming contraction to the identified specific challenge, the paradox of the common practice of supplemental feeds. Despite voicing strong support for and knowledge about EBF, participants were familiar with a variety of supplemental feeding practices in their communities. In place of universal condemnation of all supplemental feeding, policy makers might consider strategies to address the most potentially dangerous of these practices.


Prevalence and predictors of tuberculosis coinfection among HIV-seropositive patients attending the Aminu Kano Teaching Hospital, northern Nigeria.
The HIV/AIDS epidemic has been accompanied by a severe epidemic of tuberculosis (TB), although the prevalence of coinfection is largely unknown, especially in developing countries, including Nigeria. The aim of this study was to determine the prevalence and predictors of TB coinfection among HIV-seropositive Nigerians. The case files of HIV/AIDS patients attending Aminu Kano Teaching Hospital, Nigeria from January to December 2006 were reviewed. A total of 1320 HIV/AIDS patients had complete records and were reviewed, among which 138 (10.5%) were coinfected with TB (95% CI, 8.9% to 12.2%). Pulmonary TB was diagnosed in 103 (74.6%) patients, among whom only 18 (17.5%) were sputum-positive. Fifty (36.2%) coinfected patients had some type of extrapulmonary TB (EPTB); 15 had both pulmonary TB and EPTB. Among the 35 patients with EPTB only, 20 (57.1%) had abdominal TB, 5 (14.3%) had TB adenitis, 5 (14.3%) had spinal TB, 3 (8.6%) were being monitored for tuberculous meningitis, and 1 (2.9%) each had renal TB and tuberculous adrenalitis. The highest prevalence of TB, 13.7% (n = 28), was seen among patients aged 41-50 years. TB coinfection was significantly associated with marital status, WHO clinical stage, and CD4 count. Marital status (OR, 2.1; 95% CI, 1.28-3.59; P = 0.04), WHO clinical stage at presentation (4.81; 1.42-8.34; P = 0.001), and baseline CD4 count (2.71; 1.51-6.21; P = 0.02) remained significant predictors after adjustment for confounding. The moderately high prevalence of TB among HIV-seropositive patients underscores the urgent need for strategies that lead to rapid identification and treatment of coinfection with active or latent TB.


Dydrogesterone: metabolism in man.
An investigation of the urinary metabolites of the oral progestational agent dydrogesterone in healthy women of childbearing age is reported. The drug was administered in 3H-labelled form and the urine of the first 8 h, containing on average 38% of the radioactivity administered, was used as the source of the metabolites. It was fortified with urine collected during the first 8 h of a similar study with nonlabelled dydrogesterone. After enzymatic hydrolysis of conjugated metabolites, 43 different chemical species were isolated by means of extraction, followed by column and thin layer chromatography. Three of these metabolites, constituting about 70% of the urinary radioactivity, were positively identified as 20 alpha-hydroxy-9 beta, 10 alpha-pregna-4, 6-diene-3-one (52%), 21-hydroxy-9 beta, 10 alpha-pregna-4, 6-diene-3, 20-dione (18%) and 16 alpha-hydroxy-9 beta, 10 alpha-4, 6-diene-3, 20-dione (1%). Of the remainder, 20 (13%) were tentatively characterized as various products of oxidative attack, all probably having the 4, 6-diene-3-one configuration intact. It is concluded that the 4, 6-diene-3-one configuration is metabolically stable in combination with the 9 beta, 10 alpha configuration. This finding may explain why dydrogesterone, in contrast no progesterone, is orally effective, and lacks estrogenic properties.


Using 17O solid-state NMR and first principles calculation to characterise structure and dynamics in inorganic framework materials.
The use of solid-state (17)O NMR to determine local chemical environment and to characterise oxygen dynamics is illustrated in studies of zirconium tungstate, ZrW(2)O(8), and tungsten oxide, WO(3). Simple 1D magic-angle spinning (MAS) NMR allows the chemical environments in ZrW(2)O(8) to be readily characterised, and the use of a combination of one- and two-dimensional experiments to characterise oxygen dynamics in its cubic phase is reviewed. Combining local information about structure and dynamics from NMR with long-range structural information from diffraction allows a comprehensive picture of the material to be developed. Recent work is described that uses first principles calculation of NMR parameters to probe subtle asymmetries in the WO(6) octahedra that form the structural motif in WO(3). NMR is shown to be a highly sensitive probe of local structure, allowing different models derived from high-quality neutron diffraction studies to be distinguished. The density functional theory (DFT) calculations allow clear correlations between (17)O chemical shifts and distortions of the structure to be established.


Cytochrome P-450 activity in human leiomyoma and normal myometrium.
Variations in cytochrome P-450 levels may influence the responsiveness of uterine and breast tissue as well as carcinomas to endocrine therapy and may be of particular importance with agents such as tamoxifen (Nolvadex) where hydroxylation is known to alter therapeutic activities. Therefore, a sensitive spectrophotometric assay of cytochrome P-450 levels in reproductive tissue microsomes was developed to measure cyclohexane hydroxylase activity. Cyclohexane served as a substrate for several forms of cytochrome P-450. Human uterine leiomyomas (uterine fibroid tumor) contained significantly higher (p less than 0.01) cytochrome P-450 activity than adjacent normal myometrium. Specific activities for both leiomyomas (2.87 +/- 0.26 nmol/min/mg) and normal myometrium (1.60 +/- 0.11 nmol/min/mg) were in the range of those observed for untreated rabbit liver microsomes (1 to 3 nmol/min/mg). The contribution of smooth muscle in the specimen, the phase of the menstrual cycle, and the clinical diagnosis did not influence the level of cytochrome P-450 activity.


In-center hemodialysis absenteeism: prevalence and association with outcomes.
The aim of this study was to determine the rate of missed treatments among hemodialysis (HD) patients, and the association between treatment nonadherence and clinical outcomes. The data used in this study were based on electronic medical records and Medicare claims. This is a retrospective, observational study. HD patients miss 9.9% of all treatments. Approximately half of the missed treatments are due to observable medical events, predominantly hospitalizations, while half result from nonadherence ("absence"). A single absence is associated with a 1.4-fold greater risk of hospitalization, and a 2.2-fold greater risk of death in the subsequent 30 days. Treatment nonadherence is common among HD patients and is associated with adverse outcomes. Interventions that improve adherence may improve patient health and reduce costs.


Solitary adult myofibroma of the mandible simulating an odontogenic cyst.
The solitary adult myofibroma is a rare lesion but has a predilection for the head and neck. Intraosseous lesions are common in childhood but uncommon in adults. The lesion is considered to be completely benign but there is the potential for it being confused with more aggressive spindle cell tumours. Histologically it is characterised by two cell types arranged in a biphasic pattern; namely centrally positioned small rounded cells with pale staining nuclei and eosinophilic cytoplasm and peripherally elongated spindle cells. A case is reported of a 34-year-old female patient who presented with a solitary myofibroma in the lower third molar region of the mandible which clinically and radiographically simulated an odontogenic cyst.


A novel de novo TMEM63A variant in a patient with severe hypomyelination and global developmental delay.
Heterozygous variants in TMEM63A have been recently identified as the cause of infantile-onset transient hypomyelination. To date, four TMEM63A variants have been reported in five patients. These patients exhibited favorable clinical course, developmental progress, and completion of myelination. The patient was a 5-year-old girl with severe global developmental delay, absent speech, no turning over, no gazing, hypotonia, and daily episodes of autonomic seizures. Brain MRI showed hypomyelination of deep and subcortical white matter that appeared hyperintense in T2-weighted imaging from 2 months of age and that showed no change at 4 years of age. Exome sequencing of the patient and her parents revealed a novel de novo missense variant, NM_014698.3:c.1658G>T, p.(Gly553Val), in the TMEM63A gene, which was confirmed by Sanger sequencing. The variant has not been registered in public databases, and it substitutes a highly conserved glycine residue located in a pore-lining transmembrane helix. No other candidate variants were identified. Although TMEM63A variants are generally thought to cause transient hypomyelination with favorable developmental progress, identification of a de novo TMEM63A variant in our patient suggests that the TMEM63A-related clinical spectrum is broad and includes severe developmental delay with seizures.


Follow-up of patients subjected to direct and indirect pulp capping of young permanent teeth. A retrospective study.
A retrospective study of the success rate of direct pulp capping (DPC) and indirect pulp capping (IPC) was carried out in children between 6-14 years-old, considering separately primary caries or caries affecting teeth with molar incisor hypomineralization (MIH). Data were collected in a dental public health service. Following the inclusion criteria, 232 treatments were analyzed. Success was defined by the presence of a functional tooth without clinical signs or symptoms of pulpal or periapical disease. The success rate was correlated to patient gender, the affected tooth and the indication of therapy using the chi-squared and Fisher exact test. The success time related to treatment type was evaluated through the Mann-Whitney test. The IPC and DPC success rate was 99.4%, and 84.6%, respectively (p = .01). Success was significantly lower when caries affected teeth with MIH than when caries affected teeth without MIH (p = .01). The mean survival for DPC and IPC was 14.07 ± 1.30 and 15.98 ± 0.80 months, respectively (p = .07). When caries were located in teeth that were not affected by MIH, IPC was significantly more successful than DPC, but did not differ significantly when caries were placed in teeth with MIH. Key points Minimally invasive therapy is a successful approach for decayed young permanent teeth. The success of IPC was greater than the success of DPC wen caries was placed in teeth not affected by MIH. In teeth not affected by MIH the success of DPC or IPC did not differ significantly.


Inflammatory Bowel disease promote oral cancer and pharyngeal cancer: new evidence of Mendelian randomization.
Evidence from observational studies shows that inflammatory bowel disease (IBD) [comprising ulcerative colitis (UC) and Crohn's disease (CD)] is a risk factor to Oral cavity and pharyngeal cancer (OC&PC) [comprising Oral cavity cancer (OCC) and Oropharyngeal cancer (OPC)], but it is unclear whether these diseases have potential causality. We aimed to explore the causal relationship between IBD and OC&PC. A mendelian randomized (MR) study was performed to estimate the causal relationship between IBD and OC&PC. The potential causal relationship was statistically significant between IBD and OCC (OR = 1.14, 95% confidence interval (CI): 1.02-1.27, p = .02), UC and OCC (OR = 1.13, 95% CI: 1.01-1.27, p = .03), respectively. There was a universal null effect of IBD on OC&PC (IBD: OR = 1.01, 95%CI: 0.93-1.10, p = .74; UC: OR = 1.00, 95%CI: 0.92-1.10, p = .94; CD: OR = 1.02, 95%CI: 0.94-1.09, p = .69), and IBD on OPC (IBD: OR = 0.93, 95%CI: 0.81-1.06, p = 0.26; UC: OR = 0.90, 95%CI: 0.79-1.03, p = .12; CD: OR = 1.04, 95%CI: 0.94-1.15, p = .44). MR analyses support new evidence indicating there may be a positive causal effect of IBD (including UC) on OCC. Further investigation of the potential biological mechanisms is necessary.


Antimicrobial resistance and genetic diversity of Enterococcus faecalis from yolk sac infections in broiler chicks.
Despite restrictions on the use of antibiotics in poultry, the percentage of multidrug resistant bacteria, isolated from both adult birds and chicks, remains high. These bacteria can spread between countries via hatching eggs or chicks. Antibiotic resistant bacteria can also pose a threat to hatchery and farm workers or to consumers of poultry. The aim of the study was to perform a phenotypic and genotypic analysis of the drug resistance of E. faecalis isolates from yolk sac infections in broiler chicks from Poland and the Netherlands and to determine their genetic diversity. The tests revealed resistance to antibiotics from category D, that is, tetracycline (69.7%); category C - lincomycin (98.7%), erythromycin (51.3%), aminoglycosides (high-level streptomycin and kanamycin resistance - 10.5% and 3.95%, respectively), and chloramphenicol (7.9%); and category B - ciprofloxacin (25% with resistance or intermediate resistance). No resistance to penicillin, ampicillin, high-level gentamicin, tigecycline, or linezolid was noted. Various combinations of the erm(B), tet(M), tet(L), tet(O), ant(6)-Ia, aph(3')-IIIa, ant(4')-Ia, cat, and msr(A/B) genes were detected in all isolates (irrespective of the drug-resistance phenotype). Among isolates that carried the tet(M) and/or the tet(L) gene, 28% also had the Int-Tn gene, in contrast with isolates possessing tet(O). There were 28 sequence types and 43 PFGE restriction patterns. About 60% of isolates were of sequences types ST59, ST16, ST116, ST282, ST36, and ST82. Nine new sequence types were shown (ST836-ST844). In conclusion, broiler chicks can be a source of drug-resistant sequence types of E. faecalis that are potentially hazardous for people and animals. Restrictive programs for antibiotic use in broiler breeding flocks should be developed to decrease drug resistance in day-old chicks and reduce economic losses during rearing.


L-tryptophan ingestion does not induce progressive systemic sclerosis (scleroderma).
A retrospective analysis of 153 patients suffering from progressive systemic sclerosis failed to show any correlation between either onset or worsening of the disease and L-tryptophan ingestion. Only one woman developed typical acrosclerosis without signs of the eosinophilia-myalgia syndrome during exposure to the drug.


Pelvic and mammary Burkitt-like lymphoma with simultaneous development of leukemia.
The authors report the case of a young nun who came to their attention because of the simultaneous development of mammary and pelvic lymphoblastic lymphoma and acute leukemia showing aspects typical of nonendemic Burkitt's lymphoma. The rapid tumoral growth and the equally rapid spontaneous cell lysis led to severe renal insufficiency and metabolic acidosis which were ultimately the cause of death. Given the rareness of the clinical situation and the diagnostic problems involved, the authors examined the literature dealing with this subject.


Double arch impressions for simplified restorative dentistry.
Use of a double arch, closed-mouth technique as described in this article enables dentists to obtain cast and die placement in centric occlusion in a single step. Complete closure of the teeth is mandatory for registering the interocclusal relationship correctly. The accuracy of the interocclusal relationship is more likely because of the decreased number of steps in the technique. Several different trays are available for the double arch impression technique. The more rigid trays buttress the impression material and reduce the possibilities of distortion in the final impression. Several variations in the double arch impression technique are possible, and tray selection is dependent on the demands of each patient.


The diversity of microorganisms associated with Acromyrmex leafcutter ants.
Molecular biological techniques are dramatically changing our view of microbial diversity in almost any environment that has so far been investigated. This study presents a systematic survey of the microbial diversity associated with a population of Acromyrmex leafcutter ants. In contrast to previous studies on social insects, which targeted specific groups of symbionts occurring in the gut (termites, Tetraponera ants) or in specialised cells (Camponotus ants) the objective of our present study was to do a total screening of all possible micro-organisms that can be found inside the bodies of these leafcutter ants. We amplified, cloned and sequenced SSU rRNA encoding gene fragments from 9 microbial groups known to have insect-associated representatives, and show that: (1) representatives of 5 out of 9 tested groups are present, (2) mostly several strains per group are present, adding up to a total of 33 different taxa. We present the microbial taxa associated with Acromymex ants in a phylogenetic context (using sequences from GenBank) to assess and illustrate to which known microorganisms they are closely related. The observed microbial diversity is discussed in the light of present knowledge on the evolutionary history of Acromyrmex leafcutter ants and their known mutualistic and parasitic symbionts. The major merits of the screening approach documented here is its high sensitivity and specificity, which allowed us to identify several microorganisms that are promising candidates for further study of their interactions with Acromyrmex leafcutter ants or their gardens.


beta-Actin messenger RNA localization and protein synthesis augment cell motility.
In chicken embryo fibroblasts (CEFs), beta-actin mRNA localizes near an actin-rich region of cytoplasm specialized for motility, the lamellipodia. This localization is mediated by isoform-specific 3'-untranslated sequences (zipcodes) and can be inhibited by antizipcode oligodeoxynucleotides (ODNs) (Kislauskis, E.H., X.-C. Zhu, and R.H. Singer. 1994. J. Cell Biol. 127: 441-451). This inhibition of beta-actin mRNA localization resulted in the disruption of fibroblast polarity and, presumably, cell motility. To investigate the role of beta-actin mRNA in motility, we correlated time-lapse images of moving CEFs with the distribution of beta-actin mRNA in these cells. CEFs with localized beta-actin mRNA moved significantly further over the same time period than did CEFs with nonlocalized mRNA. Antizipcode ODN treatment reduced this cell translocation while control ODN treatments showed no effect. The temporal relationship of beta-actin mRNA localization to cell translocation was investigated using serum addition to serum-deprived cultures. beta-actin mRNA was not localized in serum-deprived cells but became localized within minutes after serum addition (Latham, V.M., E.H. Kislauskis, R.H. Singer, and A.F. Ross. 1994. J. Cell Biol. 126:1211-1219). Cell translocation increased over the next 90 min, and actin synthesis likewise increased. Puromycin reduced this cell translocation and blocked this induction in cytosolic actin content. The serum induction of cell movement was also inhibited by antizipcode ODNs. These observations support the hypothesis that beta-actin mRNA localization and consequent protein synthesis augment cell motility.


The cardiac arrest survival score: A predictive algorithm for in-hospital mortality after out-of-hospital cardiac arrest.
Out-of-hospital cardiac arrest (OHCA) is associated with high mortality. Current methods for predicting mortality post-arrest require data unavailable at the time of initial medical contact. We created and validated a risk prediction model for patients experiencing OHCA who achieved return of spontaneous circulation (ROSC) which relies only on objective information routinely obtained at first medical contact. We performed a retrospective evaluation of 14,892 OHCA patients in a large metropolitan cardiac arrest registry, of which 3952 patients had usable data. This population was divided into a derivation cohort (n = 2,635) and a verification cohort (n = 1,317) in a 2:1 ratio. Backward stepwise logistic regression was used to identify baseline factors independently associated with death after sustained ROSC in the derivation cohort. The cardiac arrest survival score (CASS) was created from the model and its association with in-hospital mortality was examined in both the derivation and verification cohorts. Baseline characteristics of the derivation and verification cohorts were not different. The final CASS model included age >75 years (odds ratio [OR] = 1.61, confidence interval [CI][1.30-1.99], p < 0.001), unwitnessed arrest (OR = 1.95, CI[1.58-2.40], p < 0.001), home arrest (OR = 1.28, CI[1.07-1.53], p = 0.008), absence of bystander CPR (OR = 1.35, CI[1.12-1.64], p = 0.003), and non-shockable initial rhythm (OR = 3.81, CI[3.19-4.56], p < 0.001). The area under the curve for the model derivation and model verification cohorts were 0.7172 and 0.7081, respectively. CASS accurately predicts mortality in OHCA patients. The model uses only binary, objective clinical data routinely obtained at first medical contact. Early risk stratification may allow identification of more patients in whom timely and aggressive invasive management may improve outcomes.


Renal Function: Guardian of Immune Homeostasis.
Microbial cell components from the intestinal microbiota spread systemically in a healthy host, posing the threat of inappropriate immune activation. In this issue of Immunity, Troha et al. identify a key role for renal function in the clearance of circulating Lys-type peptidoglycan, revealing a mechanism that keeps these inflammatory triggers in check to maintain systemic immune homeostasis.


Machine learning to predict the occurrence of bisphosphonate-related osteonecrosis of the jaw associated with dental extraction: A preliminary report.
The aim of this study was to build and validate five types of machine learning models that can predict the occurrence of BRONJ associated with dental extraction in patients taking bisphosphonates for the management of osteoporosis. A retrospective review of the medical records was conducted to obtain cases and controls for the study. Total 125 patients consisting of 41 cases and 84 controls were selected for the study. Five machine learning prediction algorithms including multivariable logistic regression model, decision tree, support vector machine, artificial neural network, and random forest were implemented. The outputs of these models were compared with each other and also with conventional methods, such as serum CTX level. Area under the receiver operating characteristic (ROC) curve (AUC) was used to compare the results. The performance of machine learning models was significantly superior to conventional statistical methods and single predictors. The random forest model yielded the best performance (AUC = 0.973), followed by artificial neural network (AUC = 0.915), support vector machine (AUC = 0.882), logistic regression (AUC = 0.844), decision tree (AUC = 0.821), drug holiday alone (AUC = 0.810), and CTX level alone (AUC = 0.630). Machine learning methods showed superior performance in predicting BRONJ associated with dental extraction compared to conventional statistical methods using drug holiday and serum CTX level. Machine learning can thus be applied in a wide range of clinical studies.


A case of successful treatment of a patient with hyperglycemia of 2700 mg/dL.
A 34-year-old man with obesity who was an avid consumer of soft drinks was found in a coma after complaining of a poor physical condition for a few days. On arrival, he had hyperglycemia of 2700 mg/dL, coma, shock, sepsis, aspiration pneumonia, acute renal failure, acute pancreatitis, liver dysfunction, and systemic mycosis. The rapid infusion of a large volume of isotonic saline, insulin, antibiotics, and ulinastatin was performed, and mechanical ventilation was applied. The treatment was complicated by transient hypernatremia resulting from osmostasis, which gradually decreased. He demonstrated transient decerebrate posturing upon stimulation; however, he became conscious within a week of admission, and his associated diseases also improved. After correcting his hyperglycemia, the patient was discharged on foot. We report our case of a patient with hyperglycemia of 2700 mg/dL, which was the highest value reported in the English literature. During the correction of the hyperglycemia, transient hypernatremia occurred to prevent abrupt decrease in osmolality, which thus resulted in cell swelling.


Computational insight into the anticholinesterase activities and electronic properties of physostigmine analogs.
Aim: Alzheimer's disease (AD) is known to be themajor cause of dementia among the elderly. The structural properties and binding interactions of the AD drug physostigmine (-)-phy, and its analogues (-)-hex and (-)-phe and (+)-phe, were examined, as well as their impact on the conformational changes of two different AD target enzymes AChE and BChE. Materials & methods: The conformational changes were studied using molecular dynamics and structural properties using Quantum mechanics. Results & conclusions: The binding free energy (ΔGbind) and the change in the free energy surface (FES) computed from the funnel metadynamics (FMD) simulation, both support the idea that inhibitors (-)-phe and (-)-hex have better binding activities toward enzyme AChE, and that (-)-phe is stronger in binding than the present AD drug (-)-phy.


Gastric emptying of lipids after ingestion of a solid-liquid meal in humans.
We measured gastric emptying of fat and water from a mixed meal in 6 healthy volunteers using a technique of duodenal perfusion that included phase-specific, nonabsorbable markers. Pairs of water-soluble markers (polyethylene glycol 4000 and phenolsulfonphthalein) and (equivalent) lipid-soluble ([14C]sucrose octaoleate and [3H]glycerol triether) markers were used. For each pair, one marker was used to label the corresponding component of the meal (water of fat), and the other was perfused into the duodenum as a recovery marker. The perfusion method was validated by comparing the amount of marker emptied, as estimated by marker dilution, with the amount obtained by complete aspiration of intestinal contents proximal to an occlusive balloon. Throughout the postprandial period, ratios of lipid marker to total fatty acids in the stomach remained constant, indicating a stable relationship of meal marker to total lipids. Water always emptied faster than did lipids, substantial amounts of which were still in the stomach after 6 h. There was close agreement between recovery of fat proximal to the balloon and calculations of lipid emptying from duodenal perfusion, supporting the validity of the marker dilution technique for the gastric emptying of lipids. This study demonstrates that when ingested as part of an ordinary mixed meal, water and fat have different patterns of emptying, water leaving the stomach first.


Less is more: biological effects of NiSe2/rGO nanocomposites with low dose provide new insight for risk assessment.
Nickel selenide nanomaterials (NiSe2 NMs) with different vacancies demonstrated high catalytic activity as electrocatalyst in oxygen evolution reaction. As the growing needs of the industrial applications in electrocatalyst, the increased occupational exposure and environmental releasing of NMs would be unavoidable. While, much efforts have been made to evaluate the ecological safety of such engineered NMs at unrealistically high concentrations, failed to provide the comprehensively guideline for exposure thresholds. To supplement the current knowledge gap, we testified the cytotoxicity of NiSe2/rGO nanocomposites with different surface defects under more realistic exposure mode. Compared with the short-term exposure and repetitive exposure, rat lung macrophages exhibited the augmented oxidative stress, dysfunction of mitochondria, damage of DNA and disorder of calcium homeostasis under the long-term NiSe2/rGO exposure. Noteworthily, no significant differences could be found between the NiSe2/rGO with different surface defects, indicated that the defect type of NMs were not the accurate predictor for real risk assessment. Collectively, the study provided the real potential toxic effects and exposure thresholds of NMs that might be highly possible industrial produced, and appealed the new insight for risk assessments of engineered NMs under the long-term exposure, which exhibited difference from the traditional evaluation of short-term and repetitive exposure.


Real-world outcomes of combined therapy of photodynamic therapy with anti-vascular endothelial growth factor for polypoidal choroidal vasculopathy.
To describe the real-world outcomes of photodynamic therapy (PDT) as a rescue therapy in eyes with polypoidal choroidal vasculopathy (PCV) refractory to anti-vascular endothelial growth factor (VEGF) monotherapy in a British cohort of patients. This is a retrospective chart review of 53 eyes with PCV. Based on the timing of PDT, the eyes were stratified into two groups (9 in the Initial-PDT group, 44 in the Deferred group). The number of anti-VEGF injections/year and the best corrected visual acuity (BCVA) before and after PDT were analysed. Multivariate regression model was created to identify factors predictive of visual outcome and treatment burden after PDT. The Deferred group received a mean of 9.4 injections/year but significantly reduced to 7.2 after PDT (p < 0.001). The Initial-PDT group required significantly fewer injections after PDT compared to the Deferred group (p = 0.004). The Deferred group experienced improvement in BCVA from 58.7 letters at baseline to 63.8 at 18-months follow-up (p < 0.001), but no significant increase was observed in the Initial-PDT group (p = 0.310). Better baseline BCVA is associated with higher likelihood of achieving good BCVA ≥ 70 letters after PDT (Odd Ratio=1.12, 95% CI: 1.03-1.21, p = 0.006), while increased number of anti-VEGF injections/year before PDT reduces the likelihood of easing treatment burden to ≥12 weeks apart between each injection after PDT (Odd Ratio=0.724, 95% CI: 0.58-0.91, p = 0.006). PDT as a rescue therapy is beneficial in the long-term management of PCV, particularly in eyes that had experienced a significant period of prior exposure to anti-VEGF monotherapy.


Neuropeptide Y and its receptor subtypes specifically modulate rat peritoneal macrophage functions in vitro: counter regulation through Y1 and Y2/5 receptors.
It is well documented that neuropeptide Y (NPY) exerts a wide range of biological functions through at least five NPY Y receptor subtypes (Y1-Y5), but its immunological effects only recently came into focus. Using NPY family peptides and NPY-related receptor-specific peptides as well as Y1 and Y2 receptor antagonists, we have tested which NPY Y receptors are involved in NPY-induced modulation of rat peritoneal macrophage function in vitro. NPY and PYY increased oxidative burst in phorbol myristate acetate (PMA)-stimulated macrophages involving activation of protein kinase C (PKC), and decreased it in zymosan-stimulated cells resembling inhibition of signaling pathways subsequent to binding of zymosan particles for the iC3b fragment receptor on macrophages. The combined treatment with NPY and NPY Y receptor antagonists revealed that NPY-induced potentiation of oxidative burst in PMA-stimulated cells is mediated through Y1 and Y2 receptors, while NPY-induced suppression in zymosan-stimulated cells is mediated through Y2 receptors only. NPY-related peptides differently modulated macrophage function, confirming involvement of NPY Y2 receptor in both potentiation and suppression of oxidative burst in these cells. Additionally, it was shown that NPY Y5 receptor mediated suppression of oxidative burst in PMA- and zymosan-stimulated macrophages. Taken together, the present data reveal an NPY Y1 and Y2/Y5 receptor interaction in NPY-induced modulation of macrophage functions related to inflammation.


Vancomycin Area under the Concentration-Time Curve Estimation Using Bayesian Modeling versus First-Order Pharmacokinetic Equations: A Quasi-Experimental Study.
To evaluate the efficiency of Bayesian modeling software and first-order pharmacokinetic (PK) equations to calculate vancomycin area under the concentration-time curve (AUC) estimations. Unblinded, crossover, quasi-experimental study at a tertiary care hospital for patients receiving intravenous vancomycin. Vancomycin AUC monitoring was compared using Bayesian modeling software or first-order PK equations. The primary endpoint was the time taken to estimate the AUC and determine regimen adjustments. Secondary endpoints included the percentage of vancomycin concentrations usable for AUC calculations and acute kidney injury (AKI). Of the 124 patients screened, 34 patients had usable vancomycin concentrations that led to 44 AUC estimations. Without electronic health record (EHR) integration, the time from assessment to intervention in the Bayesian modeling platform was a median of 9.3 min (quartiles Q1-Q3 7.8-12.4) compared to 6.8 min (Q1-Q3 4.8-8.0) in the PK equations group (p = 0.004). With simulated Bayesian software integration into the EHR, however, the median time was 3.8 min (Q1-Q3 2.3-6.9, p = 0.019). Vancomycin concentrations were usable in 88.2% in the Bayesian group compared to 48.3% in the PK equation group and there were no cases of AKI. Without EHR integration, Bayesian software was more time-consuming to assess vancomycin dosing than PK equations. With simulated integration, however, Bayesian software was more time efficient. In addition, vancomycin concentrations were more likely to be usable for calculations in the Bayesian group.


Quail neural crest cells cannot read positional values in the dorsal trunk feathers of the chicken embryo.
If quail neural crest cells are grafted to the chick, they migrate into the feathers of the host and produce melanin pigment. In one study, the dorsal trunk feathers of the chimaera were found to have quail-like pigment patterns. This was interpreted in terms of a positional information model. By contrast, in another study it was found that pigment patterns in the wing plumage of the chimaera bore little or no resemblance to the quail, showing instead a rather uniform, dark pigmentation. This was interpreted in terms of a prepattern in the ectoderm. This striking difference in results could be because the wing and trunk plumages have their pigment patterns specified in different ways. We have examined this possibility by making detailed maps of the dorsal trunk plumage of the normal quail and the quail-chick chimaera. Using this novel technique, we can accurately record the secondary pigment patterns in the embryonic down plumage. In the quail there are well-defined, longitudinal stripes running down the back, whereas the chimaera shows rather uniform, dark pigment in this area. There is little or no indication of stripes and some chimaerae develop asymmetric, mottled patterns. Grafts to the cephalic region also produce uniform pigmentation with no quail-like patterning. These findings indicate that neural crest cells cannot read positional values in the feathers of another species.


Investigation of Twelve Significant Mycotoxin Contamination in Nut-Based Products by the LC-MS/MS Method.
In this study, a total of 80 peanut butter, hazelnut butter, and chocolate samples were obtained from local markets in Ankara, Turkey. These foods were analyzed for twelve toxicological important mycotoxins, such as aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), and aflatoxin G2 (AFG2); fumonisin B1 (FB1) and fumonisin B2 (FB2); ochratoxin A (OTA); sterigmatocystin (STE); deoxynivalenol (DON); zearalenone (ZON); T-2 toxin (T2); and HT-2 toxin (HT2) by the LC-MS/MS multi-mycotoxin method. In addition to this analysis, the presence of total aerobic mesophilic bacteria was investigated in the samples. The samples were analyzed microbiologically using standard procedures. Finally, the minimum and maximum levels of AFB1, AFB2, AFG1, FB2, OTA, STE, DON, ZON, T2, and HT2 in the samples were found to be 0.04-27.37 µg/kg, 0.06-6.19 µg/kg, 0.14-0.40 µg/kg, 2.73-2.93 µg/kg, 0.01-37.26 µg/kg, 0.19-2.25 µg/kg, 11.81-42.09 µg/kg, 0.03-7.57 µg/kg, 1.41-2.54 µg/kg, and 6.94-7.43 µg/kg, respectively. AFG2 and FB1 were not detected in any of the samples. The most frequently detected mycotoxins in analyzed samples were OTA (78.75%) and AFB1 (75%). In addition, total aerobic mesophilic bacteria were isolated from 53.75% of samples. Some of the tested food samples contained mycotoxins above the Turkish Food Codex maximum limit.


6-(4-Bromophenyl)-6,7-dihydro-1,3-dioxolo[4,5-g]quinolin-8(5H)-one: bilayers built from N-H...O, C-H...O and C-H...pi(arene) hydrogen bonds.
Molecules of the title compound, C(16)H(12)BrNO(3), exhibit a polarized molecular-electronic structure. A combination of one N-H...O hydrogen bond and one C-H...O hydrogen bond links the molecules into sheets, and pairs of sheets are linked into bilayers by a single C-H...pi(arene) hydrogen bond.


FIT2 organizes lipid droplet biogenesis with ER tubule-forming proteins and septins.
Lipid droplets (LDs) are critical for lipid storage and energy metabolism. LDs form in the endoplasmic reticulum (ER). However, the molecular basis for LD biogenesis remains elusive. Here, we show that fat storage-inducing transmembrane protein 2 (FIT2) interacts with ER tubule-forming proteins Rtn4 and REEP5. The association is mainly transmembrane domain based and stimulated by oleic acid. Depletion of ER tubule-forming proteins decreases the number and size of LDs in cells and Caenorhabditis elegans, mimicking loss of FIT2. Through cytosolic loops, FIT2 binds to cytoskeletal protein septin 7, an interaction that is also required for normal LD biogenesis. Depletion of ER tubule-forming proteins or septins delays nascent LD formation. In addition, FIT2-interacting proteins are up-regulated during adipocyte differentiation, and ER tubule-forming proteins, septin 7, and FIT2 are transiently enriched at LD formation sites. Thus, FIT2-mediated nascent LD biogenesis is facilitated by ER tubule-forming proteins and septins.


Effects of single and fractionated low-dose irradiation on vascular endothelial cells.
An increasing number of epidemiological studies suggest that chronic low-dose irradiation increases the risk of atherosclerosis. We evaluated and compared the in vitro biological effects of both single and fractionated low-doses of X-ray irradiation on endothelial cells. Human umbilical vein endothelial cells (HUVECs) were irradiated with X-rays, with single doses of 0.125, 0.25 and 0.5 Gy or fractionated doses of 2 × 0.125 Gy and 2 × 0.25 Gy, with 24 h interfraction interval. Survival, apoptosis, reactive oxygen species (ROS) production, nuclear factor-κB (NF-κB) activation, intercellular adhesion molecule-1 (ICAM-1) expression, HUVEC adhesiveness and DNA damage were investigated. We did not observe any effect on viability and apoptosis. Both single and fractionated doses induced ROS generation, NF-κB activation, ICAM-1 protein expression and HUVEC adhesiveness, but only fractionated doses increase significantly ICAM-1 mRNA. The effects measured after fractionated dose result always higher than those induced by the single dose. Moreover, we observed that DNA double strand break (DSB), visualized with γ-H2AX foci, is dose-dependent and that the kinetics of γ-H2AX foci is not affected by fractionated doses. We showed that single and fractionated low-dose irradiations with low energy X-rays do not affect cell viability and DNA repair. Interestingly, the greater increase of ICAM-1 surface exposure and endothelial adhesiveness observed after fractionated irradiation, suggests that fractionated low-doses may accelerate chronic vascular inflammation, from which the atherosclerotic process can arise.


Selection of Phlebotomus papatasi (Diptera: Psychodidae) lines susceptible and refractory to Leishmania major infection.
Variation in susceptibility to infection with cultured promastigotes of Leishmania major was detected among 3 different geographic strains (ISRAEL, EGYPT, and INDIA) of the sand fly Phlebotomus papatasi. The ISRAEL strain showed the greatest susceptibility and was chosen for subsequent genetic selection experiments. After 13 generations of genetic selection in this strain, a stable refractory line was obtained in which only 7.5% of the insects could be infected. A highly susceptible line was also obtained in 5 generations of selection. Infection rates in the susceptible line were stabilized at about 94%. A description of the selection process and characteristics of the 2 lines are given.


Repurposing QuantiFERON for Detection of Neutralizing Interferon-γ Autoantibodies in Patients With Nontuberculous Mycobacterial Infections.
Nontuberculous mycobacterial infections due to autoantibodies targeting interferon-γ are an emerging medical problem. However, case finding is hampered due to highly complex diagnostic procedures not available in routine laboratories. We show that QuantiFERON assays can be exploited as a simple screening tool that may facilitate adequate and timely treatment.


A study protocol for MindMoves: A lifestyle physical activity and cognitive training intervention to prevent cognitive impairment in older women with cardiovascular disease.
Cognitive impairment (CI) and cardiovascular disease (CVD) disproportionately affect women compared to men, and CVD increases risk of CI. Physical activity and cognitive training can improve cognition in older adults and may have additive or synergistic effects. However, no combined intervention has targeted women with CVD or utilized a sustainable lifestyle approach. The purpose of the trial is to evaluate efficacy of MindMoves, a 24-week multimodal physical activity and cognitive training intervention, on cognition and serum biomarkers in older women with CVD. Three serum biomarkers (brain-derived neurotrophic factor [BDNF], vascular endothelial growth factor [VEGF], and insulin-like growth factor 1 [IGF-1]) were selected as a priori hypothesized indicators of the effects of physical activity and/or cognitive training on cognition. The study design is a randomized controlled trial with a 2 × 2 factorial design, to determine independent and combined efficacies of Mind (tablet-based cognitive training) and Move (lifestyle physical activity with goal-setting and group meetings) on change in cognition (primary outcome) and serum biomarkers (secondary outcomes). We will recruit 254 women aged ≥65 years with CVD and without CI from cardiology clinics. Women will be randomized to one of four conditions: (1) Mind, (2) Move, (3) MindMoves, or (4) usual care. Data will be obtained from participants at baseline, 24, 48, and 72 weeks. This study will test efficacy of a lifestyle-focused intervention to prevent or delay cognitive impairment in older women with CVD and may identify relevant serum biomarkers that could be used as early indicators of intervention response.


Guidance on the limits to the number of embryos to transfer: a committee opinion.
Based on American Society for Reproductive Medicine (ASRM) and Society for Assisted Reproductive Technology data available through 2014, ASRM's guidelines for the limits on the number of embryos to be transferred in in vitro fertilization (IVF) cycles have been further refined in continuing efforts to promote singleton gestation and reduce the number of multiple pregnancies. This version replaces the document titled Criteria for number of embryos to transfer: a committee opinion that was published most recently in August of 2013 (Fertil Steril 2013;99:44-6).


Neurologıcal Complıcatıons and Potentıal Effects of Covıd-19: Symptoms and Conceıvable Mechanısms.
A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in December 2019 in Wuhan, China. The new coronavirus disease (COVID-19) was declared a global pandemic by the World Health Organization (WHO) in March 2020. SARS-CoV-2 can invade the nervous system aside from infecting the respiratory system as its primary target. The most common nervous system symptoms of COVID-19 are stated as headache, myalgia, fatigue, nausea, vomiting, sudden and unexplained anosmia, and ageusia. More severe conditions such as encephalomyelitis, acute myelitis, thromboembolic events, ischemic stroke, intracerebral hemorrhage, Guillain-Barré-syndrome, Bell's palsy, rhabdomyolysis, and even coma have also been reported. Cohort studies revealed that neurological findings are associated with higher morbidity and mortality. The neurological symptoms and manifestations caused by SARS-CoV-2 and COVID-19 are examined and summarized in this article.


Oral and oropharyngeal cancer.
Oral and oropharyngeal cancer (OPC) is a complex and often relentless malignancy prone to local invasion and dissemination. Despite advances in understanding of the disease and improved therapeutic interventions, it continues to be diagnosed at an advanced stage and the survival rate remains poor. The financial cost of treating OPC may be the highest of all cancers in the United States and survivors often experience major detriments to quality of life. Major risk factors for OPC are tobacco, alcohol, areca nut, and human papillomavirus infection. This article updates medical practitioners on the causes, presentation, diagnosis, and management of OPC.


Extremes of age are associated with differences in the expression of selected pattern recognition receptor genes and ACE2, the receptor for SARS-CoV-2: implications for the epidemiology of COVID-19 disease.
Older aged adults and those with pre-existing conditions are at highest risk for severe COVID-19 associated outcomes. Using a large dataset of genome-wide RNA-seq profiles derived from human dermal fibroblasts (GSE113957) we investigated whether age affects the expression of pattern recognition receptor (PRR) genes and ACE2, the receptor for SARS-CoV-2. Extremes of age are associated with increased expression of selected PRR genes, ACE2 and four genes that encode proteins that have been shown to interact with SAR2-CoV-2 proteins. Assessment of PRR expression might provide a strategy for stratifying the risk of severe COVID-19 disease at both the individual and population levels.


Predictive power of the ESUR scoring system for prostate cancer diagnosis verified with targeted MR-guided in-bore biopsy.
This study evaluates the diagnostic value of the ESUR scoring system (PI-RADS) regarding prostate cancer detection using MR-guided in-bore biopsies (IB-GB) as the reference standard. 566 lesions in 235 consecutive patients (65.7 ± 7.9 years, PSA 9.9 ± 8.5 ng/ml) with a multiparametric (mp)-MRI (T2WI, DWI, DCE) of the prostate at 3T were scored using the PI-RADS scoring system. PI-RADS single (PSsingle), summed (PSsum), and overall (PSoverall) scores were determined. All lesions were histologically verified by IB-GB. Lesions with a PSsum below 9 contained no prostate cancer (PCa) with Gleason score (GS) ≥ 4+3=7. A PSsum of 13-15 (PSoverall V) resulted in 87.8% (n=108) in PCa and in 42.3% (n=52) in GS ≥ 4+3=7. Transition zone (TZ) lesions with a PSsum of 13-15 (PSoverall V) resulted in 76.3% (n=36) in PCa and in 26.3% (n=10) in GS ≥ 4+3=7, whereas for peripheral zone (PZ) lesions cancer detection rate at this score was 92.9% (n=79) and 49.4% (n=42) for GS ≥ 4+3=7. Using a threshold of PSsum ≥ 10, sensitivity was 86.0%, and negative predictive value (NPV) was 86.2%. For higher grade PCa sensitivity was 98.6%, and NPV was 99.5%. A PSsum below 9 excluded a higher grade PCa, whereas lesions with a PSsum ≥ 13 (PSoverall V) represented in 88% PCa, and in 42% higher grade PCa. The PSsum or PSoverall demonstrated a better diagnostic value for PZ lesions with higher detection rates for higher grade PCa compared to TZ lesions.


An ensemble of interconverting conformations of the elemental paused transcription complex creates regulatory options.
Transcriptional pausing underpins the regulation of cellular RNA synthesis, but its mechanism remains incompletely understood. Sequence-specific interactions of DNA and RNA with the dynamic, multidomain RNA polymerase (RNAP) trigger reversible conformational changes at pause sites that temporarily interrupt the nucleotide addition cycle. These interactions initially rearrange the elongation complex (EC) into an elemental paused EC (ePEC). ePECs can form longer-lived PECs by further rearrangements or interactions of diffusible regulators. For both bacterial and mammalian RNAPs, a half-translocated state in which the next DNA template base fails to load into the active site appears central to the ePEC. Some RNAPs also swivel interconnected modules that may stabilize the ePEC. However, it is unclear whether swiveling and half-translocation are requisite features of a single ePEC state or if multiple ePEC states exist. Here, we use cryo-electron microscopy (cryo-EM) analysis of ePECs with different RNA-DNA sequences combined with biochemical probes of ePEC structure to define an interconverting ensemble of ePEC states. ePECs occupy either pre- or half-translocated states but do not always swivel, indicating that difficulty in forming the posttranslocated state at certain RNA-DNA sequences may be the essence of the ePEC. The existence of multiple ePEC conformations has broad implications for transcriptional regulation.


Surgical management of thymic epithelial tumors: a retrospective review of 204 cases.
Thymic epithelial tumors consist of a series of neoplasm that differ morphologically and biologically. Management strategy for these tumors remains controversial. We retrospectively reviewed 204 thymic epithelial tumors surgically treated during the period of 1970 to 1995, and we reexamined the histologic specimens using the current World Health Organization classification. One hundred eighty patients (88.2%) underwent complete resections, 17 (8.3%) received partial resections, and 7 (3.4%) received biopsies only. The complete resection rate of stages I and II tumors was significantly higher than stages III and IV tumors (98.2% vs 76.6%; p < 0.001). Twenty-four patients (11.8%) experienced 29 events of complications postoperatively, including 11 events of myasthenia gravis crisis. Myasthenia gravis, partial resection or biopsy, and stages III and IV were independent risk factors for postoperative complications. Fifteen patients (7.4%) died during hospital stays, including 6 patients from myasthenia crisis. Partial resection or biopsy and myasthenia gravis were independent risk factors for postoperative mortality. There were significantly more stage I and stage II cases in histologic types A, AB, and B1 tumors than in B2, B3, and C tumors (87.6% vs 26.4%; p < 0.001), and their complete resection rate was significantly higher than the latter group (98.9% vs 78.3%; p < 0.001). The 5-year and 10-year survival rates were 63.2% and 50.4%, respectively. Masaoka stages III and IV, histologic types B2, B3, and C, and incomplete resection were independent risk factors for poor prognosis. Complete resection remains the hope of cure for thymic epithelial tumors. The treatment strategy should be based on the current World Health Organization histologic classification and the Masaoka staging system.


Comprehensive investigation of antibiotic resistance gene content in cfiA-harboring Bacteroides fragilis isolates of human and animal origins by whole genome sequencing.
The emergence of multidrug resistance in Bacteroides fragilis, especially the phylogenetic lineage carrying the carbapenemase gene cfiA, represents an increasing threat to human health. However, knowledge on the diversity of the multidrug-resistant strains and the genetic elements carrying the antibiotic resistance genes (ARGs) remains limited. The objective of the study was to describe the resistome in cfiA-positive B. fragilis. A collection of cfiA-positive B. fragilis from diverse human (8 bacteremias, 15 wound infections) and animal (2 chickens, 2 pigs, 6 dogs, 3 cats) sources in Hong Kong, 2015-2017 was analysed by whole genome sequencing. In the 36 isolates, 13 distinct ARGs (total number 83, median 2, range 0-7 per isolate) other than cfiA were detected. ARGs encoding resistance to aminoglycosides, β-lactams, macrolides, sulphonamides and tetracyclines were carried by CTn341-like, CTnHyb-like, Tn5220-like, Tn4555-like and Tn613-like transposons and were detected in phylogenetically diverse isolates of different host sources. Only few ARGs encoding resistance to metronidazole and tetracyclines were localized on plasmids. In two chicken isolates, a novel transposon (designated as Tn6994) was found to be involved in the dissemination of multiple ARGs mediating resistance to multiple antibiotics, including metronidazole and linezolid that are critically important for treatment of anaerobic infections. In mating experiments, Tn6994 and the associated phenotypic resistance could be transferred to Bacteroides nordii recipient. This study illustrates the importance of transposons in the dissemination of ARGs in the cfiA-positive division of B. fragilis. One Health approach is necessary to track the dissemination of ARGs.


Influence of steroid therapy on the incidence of pericarditis and atrial fibrillation after percutaneous epicardial mapping and ablation for ventricular tachycardia.
This study evaluates the influence of 3 therapeutic approaches on the incidence of pericarditis and atrial fibrillation (AF) after percutaneous epicardial mapping and ablation for ventricular tachycardia. Eighty-five consecutive procedures (2006-2011) were retrospectively reviewed. After the first 17 procedures (20.0%), no steroids were administered. For the subsequent 30 procedures (35.3%), systemic steroids were administered intravenously or orally, whereas the last 38 procedures (44.7%) were followed by intrapericardial steroid injection. Compared with no steroids, the incidence of pericarditic chest pain was significantly reduced by intrapericardial steroids (58.8% versus 21.1%; P=0.006) but not by intravenous or oral steroids (58.8% versus 43.4%; P=0.31). There was no significant difference in the incidence of pericarditic ECG with steroids (36.8%, 30.0%, and 41.2% for intrapericardial, intravenous or oral, and none, respectively). There was a nonsignificant reduced incidence of chest pain with ECG changes with steroids (13.2%, 10.0%, and 29.4% for intrapericardial, intravenous or oral, and none, respectively). Radiofrequency applications (65.9% of procedures) did not affect the incidence of pericarditic ECG changes, pericarditic chest pain, or pericarditis (all P>0.05). In 7 (8.3%) patients with no prior history of AF, AF was documented a median 36 hours after procedure. Patients with pericarditic ECG tended to be at greater risk of AF (16.7 versus 3.6%; P=0.091). There is a high incidence of pericarditic chest pain and ECG changes after epicardial ventricular tachycardia mapping and ablation. Pericarditic chest pain is significantly decreased by intrapericardial steroids. Procedure-related AF is relatively frequent and tends to occur more commonly with pericarditic ECG changes.


Utilizing plasma drug levels and genetic testing to achieve optimal treatment response in a patient with treatment-resistant schizoaffective disorder.
We report the case of a Chinese male with schizoaffective disorder, an active smoker and a nonresponder to clozapine (600 mg daily). Therapeutic clozapine monitoring was analyzed, revealing a low concentration-dose ratio. A pharmacogenetic test showed that the patient had the CYP1A2*1F/*1F genotype, indicating an ultra-rapid clozapine metabolizer. In combination with fluvoxamine, a CYP1A2 enzyme inhibitor, clozapine plasma concentrations approached the reference range and achieved clinical improvement. This case demonstrates how pharmacogenetics can help understand the value of therapeutic drug monitoring to enhance the treatment of refractory schizoaffective disorder.


Protein synthesis inhibition and amnesia for saccharin aversion memory in rats after intra-cisternal administration of cycloheximide.
The series of experiments reported further investigated the time course relationship of the amnesia for saccharin aversion memory in rats and the inhibition of protein synthesis resulting from cycloheximide (CXM). The first experiment showed that intraventricular CXM injected rats which had been trained 7 hrs after injection had learned the contingency under a considerable inhibition of protein synthesis. However, 1 hr after CXM injection protein synthesis inhibition was 86% and yet there was no amnesia observed 24 hr after training. It was also apparent that there were no marked regional differences in the extent of protein synthesis inhibition after intraventricular CXM. Finally, intracisternal injection of CXM 1 hr before training resulted in amnesia 24 hrs after training although this effect was greater when the injection was performed 7 hr before training. The findings (1) are consistent with Day et al's [4] suggestion of maximal protein synthesis inhibition at 1 hr post CXM injection with an approximately linear decline thereafter, and (2) provide no support for the involvement of the brain stem nucleus solitarius in taste aversion learning.


Antilipidemic activity of 4-oxo-functionalized ethyl 6-chlorochroman-2-carboxylate analogs and a related tricyclic lactone in three rat models.
The synthesis of ethyl cis-6-chloro-4-hydroxychroman-2-carboxylate (IV) and 6-chloro-4-hydroxy-chroman-2-carboxylic acid lactone (V) are reported. The antilipidemic properties of these compounds in 3 rat models were compared to the activity obtained for the previously synthesized related analogs ethyl 6-chlorochroman-2-carboxylate (II), ethyl 6-chlorochromanone-2-carboxylate (III) and clofibrate (I). The biologically most interesting analog, ethyl 6-chlorochroman-2-carboyxlate (II) like clofibrate (I), was an effective antitriglyceridemic and anticholesterolemic agent in Triton WR-1339 hyperlipidemic rats, sucrose-fed hyperlipidemic rats and chow-fed normolipemic rats. Ethyl 6-chlorochromanone-2-carboxylate (III) was found to be active after 7 days of administration to sucrose-fed rats. In sucrose-fed, male Sprague-Dawley rats, the comparative effects of these analogs on various hepatic drug parameters also were carried out. Consistent with previous findings, results obtained with these compounds provide evidence showing that changes in hepatic HMG-CoA reductase activity bear no relationship to serum cholesterol lowering in the sucrose-fed model.


Nanozyme enhanced paper-based biochip with a smartphone readout system for rapid detection of cyanotoxins in water.
Cyanobacterial harmful algal blooms in freshwater systems can produce cyanotoxins, such as microcystins (MCs) and nodularins (NODs), presenting serious threats to human health and ecosystems. Required routine monitoring of cyanotoxins in water samples, as posed by U.S. EPA drinking water contaminant candidate list 5 (CCL5), demands for cost-effective, reliable and sensitive MCs/NODs detection methods. We report the development of a colorimetric paper-based immunochip assisted by nanozyme catalysis with a smartphone readout system for rapid detection of cyanotoxins in water. We show that the introduction of biorthogonal click reaction enables in situ facile self-assembly of multi-layers of peroxidase-like nanozyme onto the anti-MCs/NODs monoclonal antibody. We can detect 13 variants of MCs/NODs even in the sub-microgram per liter range with detection limit of below 0.7 μg/L and satisfactory recovery percentages between 88 and 120% in different water matrices. Our technology shows a good correlation with the well-developed ELISA technology, demonstrating its great potential applications in resource-limited or less-developed regions for on-site and large-scale screening of cyanotoxins in water environment.


Refined human artificial chromosome vectors for gene therapy and animal transgenesis.
Human artificial chromosomes (HACs) have several advantages as gene therapy vectors, including stable episomal maintenance, and the ability to carry large gene inserts. We previously developed HAC vectors from the normal human chromosomes using a chromosome engineering technique. However, endogenous genes were remained in these HACs, limiting their therapeutic applications. In this study, we refined a HAC vector without endogenous genes from human chromosome 21 in homologous recombination-proficient chicken DT40 cells. The HAC was physically characterized using a transformation-associated recombination (TAR) cloning strategy followed by sequencing of TAR-bacterial artificial chromosome clones. No endogenous genes were remained in the HAC. We demonstrated that any desired gene can be cloned into the HAC using the Cre-loxP system in Chinese hamster ovary cells, or a homologous recombination system in DT40 cells. The HAC can be efficiently transferred to other type of cells including mouse ES cells via microcell-mediated chromosome transfer. The transferred HAC was stably maintained in vitro and in vivo. Furthermore, tumor cells containing a HAC carrying the suicide gene, herpes simplex virus thymidine kinase (HSV-TK), were selectively killed by ganciclovir in vitro and in vivo. Thus, this novel HAC vector may be useful not only for gene and cell therapy, but also for animal transgenesis.


Dynamic imaging of autophagy-lysosomal pathway and autophagy function following pulmonary hypoxia/reoxygenation in vitro.
Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.


Production and Use of Gesicles for Nucleic Acid Delivery.
Over-expression of the vesicular stomatitis virus glycoprotein (VSVG) in mammalian cells can induce the formation of VSVG-pseudotyped vesicles (named "gesicles") from membrane budding. Its use as a nucleic acid delivery tool is still poorly documented. Naked-plasmid DNA can be delivered in animal cells with gesicles in presence of hexadimethrine bromide (polybrene). However, little is known about gesicle manufacturing process and conditions to obtain successful nucleic acid delivery. In this study, gesicles production process using polyethylenimine (PEI)-transfected HEK293 cells was developed by defining the VSVG-plasmid concentration, the DNA:PEI mass ratio, and the time of gesicle harvest. Furthermore, parameters described in the literature relevant for nucleic acid delivery such as (i) component concentrations in assembly mixture, (ii) component addition order, (iii) incubation time, and (iv) polybrene concentration were tested by assessing the transfection capacity of the gesicles complexed with a green fluorescent protein (GFP)-coding plasmid. Interestingly, freezing/thawing cycles and storage at + 4 °C, - 20 °C, and - 80 °C did not reduce gesicles' ability to transfer plasmid DNA. Transfection efficiency of 55% and 22% was obtained for HeLa cells and for hard-to-transfect cells such as human myoblasts, respectively. For the first time, gesicles were used for delivery of a large plasmid (18-kb) with 42% of efficiency and for enhanced green fluorescent protein (eGFP) gene silencing with siRNA (up to 60%). In conclusion, gesicles represent attractive bioreagents with great potential to deliver nucleic acids in mammalian cells.


Tamsulosin as adjunctive treatment after shockwave lithotripsy in patients with upper urinary tract stones: a systematic review and meta-analysis.
This study aimed to evaluate the efficacy of tamsulosin as an α(1)-blocker in the treatment of the renal and ureteral stones after shockwave lithotripsy (SWL). Relevant randomized controlled trials were identified by electronic and document searches of MEDLINE, EMBASE, the Chinese Biomedical Disk and the Cochrane Central Register of Controlled Trials until April 2010. No language restriction was applied. Trials were included if patients were randomized to receive either tamsulosin or standard therapy with or without placebo after SWL. The main outcome was the stone clearance rate. Two reviewers independently assessed trial quality and extracted data. Meta-analysis was conducted with Review Manager (RevMan) version 5.0. Fifteen studies involving 1326 subjects met the inclusion criteria. Study duration ranged from 2 weeks to 3 months. The pooled analysis showed a 24% [risk ratio (RR) = 1.24, 95% confidence interval (CI) 1.12 to 1.37] improvement in stone clearance tamsulosin. According to stone locations (renal, upper ureteral and lower ureteral), the pooling effects of tamsulosin were analysed, with a higher expulsion rate obtained than control (RR = 1.38, 1.83, 1.43, and 95% CI 1.17 to 1.61, 1.20 to 2.78, 1.13 to 1.81, respectively). Tamsulosin 0.4 mg had a high possibility of achieving successful outcome (RR = 1.29, 95% CI 1.14 to 1.47). In addition, a shorter expulsion time, lower analgesic requirements, fewer colic episodes and adverse effects were observed. Tamsulosin is a safe and effective therapy for renal and ureteral stones after SWL. Further, high-quality randomized trials are necessary to confirm its efficacy.


Radiofrequency coagulation with Thiersch's operation--a better palliative treatment in prolapse rectum.
Thiersch operation is an age-old popular option for treatment of complete rectal prolapse (proctodentia) in elderly patients. This procedure is simple, easy to perform, and with very rare postoperative complications of any severe nature. This is especially suitable for those patients who are unfit for extensive abdominal surgical procedures in practice today. This procedure does help in mechanically supporting and containing the prolapse. Although the procedure is not curative, the confining of the prolapse in the anal canal does ensure a relief from symptoms generated by the prolapse. Nevertheless, withdrawal of the occlusive material used is inevitably followed by recurrence of the prolapse, making the procedure a failure in achieving a prolonged relief. Looking at this drawback, we have made an addition to this conventional procedure by introducing circumferential coagulation of the subanodermal region through a radio frequency device. This results in inducing fibrosis and forming a zone of band. The added maneuver constricts the anal opening permanently and thus aids in reinforcement of the atonic sphincter. The study was carried out in 8 patients. Their average age was 57 years. All of them had complete prolapse of the rectum with associated features. Two of them were in early fifties, but they refused to undergo extensive procedures and opted for this method considered cheaper and quicker. A follow-up for a period ranging from 15 to 18 months was found satisfactory. None of the patients had any prolapse or its sequel. Not a single case was reported for any sinus or discharge from the site of incision. The incision site was well consolidated, and the knot or the thread could not be felt when seen after 12 months. The study shows that radio frequency coagulation followed by Theirsch operation for complete rectal prolapse in the elderly has been found to be relatively easy to perform. Except for the radio surgical unit, no special instrumentation is needed. The results of this procedure have been found to be gratifying in all the patients. It is felt that on appropriate selection of the patients and employing this new technique of operation would prove a boon in treatment of this problem. A further work in this direction and longer follow-up of the treated patients are bound to bring about exciting new dimensions in the field.


Phenolic profiles and bioactivities of different milling fractions of rice bran from black rice.
Phytochemicals are unevenly distributed in grain kernels and concentrated in bran fractions. However, their specific distribution in the grain bran, especially colored grains, is not clarified. This study divided rice bran from black rice into five fractions by stepwise milling to obtain BF1(outermost layer) to BF5 (the innermost layer). Each fraction accounted for approximately 2% of the whole kernel. The total content of phenolics (TPC), flavonoids (TFC), and anthocyanins (TAC) of five fractions significantly decreased from BF1 to BF5. The TPC, TFC and TAC of BF1 contribute 25.7%, 28.2%, 28.4% to the total of five fractions, respectively. HPLC analysis showed that the contents of most anthocyanin and phenolic acids compounds decreased from BF1 to BF5. Together with α-glucosidase and α-amylase inhibitory activities of BF1, the antioxidant activity was higher than those of other fractions. These results can guide the moderate processing of black rice and the utilization of its bran.


Hepatic accumulation of Hedgehog-reactive progenitors increases with severity of fatty liver damage in mice.
Progenitors regenerate fatty livers but the mechanisms involved are uncertain. The Hedgehog pathway regulates mesendodermal progenitors and modulates mesenchymal-epithelial interactions during tissue remodeling. To determine if Hedgehog signaling increases in liver progenitors during fatty liver injury, we compared expression of Hedgehog ligands and target genes across a spectrum of injury. Leptin-deficient ob/ob mice with fatty livers and their healthy lean littermates were studied before and after exposure to the hepatotoxin, ethionine. At baseline, ob/ob mice had greater liver damage than controls. Ethionine induced liver injury in both ob/ob and lean mice, with greater injury occurring in ob/ob mice. After ethionine, the ob/ob mice developed liver atrophy and fibrosis. Liver injury increased hepatic accumulation of progenitors, including ductular cells that produced and responded to Hedgehog ligands. A dose-response relationship was demonstrated between liver injury and expansion of Hedgehog-responsive progenitors. In severely damaged, atrophic livers, nuclei in mature-appearing hepatocytes accumulated the Hedgehog-regulated mesenchymal transcription factor, Gli2 and lost expression of the liver epithelial transcription factor, hepatocyte nuclear factor 6 (HNF-6). Hepatic levels of collagen mRNA and pericellular collagen fibrils increased concomitantly. Hence, fatty liver injury increases Hedgehog activity in liver progenitors, and this might promote epithelial-mesenchymal transitions that result in liver fibrosis.


Apelin stimulates glucose uptake through the PI3K/Akt pathway and improves insulin resistance in 3T3-L1 adipocytes.
Apelin, a cytokine mainly secreted by adipocytes, is closely related with insulin resistance. The underlying molecular mechanisms of how apelin affects insulin resistance, however, are poorly understood. This study aimed to investigate the effect of apelin on glucose metabolism and insulin resistance in 3T3-L1 adipocytes. After 10 ng/ml TNF-α treatment for 24 h, insulin-stimulated glucose uptake was reduced by 47% in 3T3-L1 adipocytes. Apelin treatment improved glucose uptake in a time- and dose-dependent manner. Treatment of 1,000 nM apelin for 60 min maximally augmented glucose uptake in insulin-resistant 3T3-L1 adipocytes. Furthermore, apelin pre-incubation also increased adipocytes' insulin-stimulated glucose uptake, and PI3K/Akt pathway were involved in these effects. In addition, immunocytochemistry staining and western blotting analysis indicated that apelin could increase glucose transporter 4 translocation from the cytoplasm to the plasma membrane. Apelin also increased the anti-inflammatory adipokine adiponectin mRNA expression while reducing that of pro-inflammatory adipokine interleukin-6 in insulin-resistant 3T3-L1 adipocytes. These results suggest that apelin stimulates glucose uptake through the PI3K/Akt pathway, promotes GLUT4 translocation from the cytoplasm to the plasma membrane, and modulates inflammatory responses in insulin-resistant 3T3-L1 adipocytes.


Circular RNA 0001313 Knockdown Suppresses Non-Small Cell Lung Cancer Cell Proliferation and Invasion via the microRNA-452/HMGB3/ERK/MAPK Axis.
Non-small cell lung cancer (NSCLC) seriously endangers human health. Circular RNAs (circRNAs) regulate diverse types of cancers, including NSCLC. This study investigated the possible mechanism of circ0001313 in NSCLC. Circ0001313 expression in NSCLC tissues was measured, and its correlation with clinicopathological features was analyzed. The binding relationships among circ0001313, microRNA (miR)-452 and HMGB3 were tested. The gain and loss of functions were performed to examine NSCLC cell malignant behaviors. After HMGB3 overexpression, ERK/MAPK pathway-related protein levels were detected. Subsequently, the rescue experiment was further performed using an ERK/MAPK pathway inhibitor PD98059. Abnormally elevated circ0001313 and decreased miR-452 in NSCLC cells were observed. Circ0001313 silencing or miR-452 overexpression significantly reduced NSCLC cell proliferation and invasion. Circ0001313 competitively bound to miR-452 to upregulate HMGB3, thus promoting NSCLC cell growth. HMGB3 overexpression activated the ERK/MAPK pathway to contribute to NSCLC development. We highlighted that silencing of circ0001313 blunted the ERK/MAPK pathway via the miR-452/HMGB3 axis, thereby inhibiting NSCLC cell proliferation and invasion.


Epidemiology and risk factors for corneal graft rejection.
The aim of this study was to evaluate the frequency and main risk factors for corneal graft rejection. This retrospective study included 285 eyes in 256 patients who underwent a penetrating keratoplasty (KPT) from January 1995 to December 2004. The minimum follow-up was 12 months to evaluate graft evolution. Except for complications, the follow-up was weekly, then monthly for 6 months, and ultimately quarterly during the first year. Thereafter the follow-up was performed semi-annually. Patients were informed about the functional signs for which they have to urgently consult. Immunologic rejection of the corneal graft occurred in 128 KPT in 112 patients (rejection frequency = 41%). The identified main risk factors were new vascularization of the recipient cornea over 2 or more quadrants, corneal opacity due to an infectious origin, posttraumatic corneal opacity or congenital glaucoma, graft diameter >8 mm, and therapeutic KPT. Rejection of the corneal graft is the primary cause of KPT failure. One out of 2 graft failures was due to rejection. Two criteria are unanimously recognized as risk factors for rejection: neovascularization of recipient cornea and antecedents of corneal rejection. The rejection must be treated early to not endanger graft success, which imposes a close follow-up for grafted patients.


Acute electrical stimulation for urinary incontinence.
Acute or maximal electric stimulation of the pelvic floor muscles has been used in incontinent patients who are suitable candidates for electrical stimulation, but unwilling or unable to use the anal plug electrodes. Seventeen of 20 patients had relief or improvement of their incontinence. However 5 of these 17 patients had a relapse of symptoms on follow-up, requiring a repeat treatment with acute or maximal electrical stimulation.


[The structural-functional characteristics of the rostral sections of the neocortex in Chiroptera].
Studies have been made on the connections of rostral neocortex in bats in order to reveal connections with the structures of the auditory sensory system the existence of which is indicated by evident specific responses to ultrasound in the form of synchronization reaction. It was shown that dorsolateral parts of the rostral neocortex receive topically organized projections from the thalamic nuclei VPL and VL. Connections with the auditory cortex and suprageniculate nucleus are not evident. Afferents of the medial wall of the rostral cortex originate from the thalamic nuclei MD and AM. Possible pathways of auditory afferentation to the dorso-lateral part of rostral neocortex are discussed.