Effect of knee joint icing on knee extension strength and knee pain early after total knee arthroplasty: a randomized cross-over study.
To investigate the acute effect of knee joint icing on knee extension strength and knee pain in patients shortly after total knee arthroplasty. A prospective, single-blinded, randomized, cross-over study. A fast-track orthopaedic arthroplasty unit at a university hospital. Twenty patients (mean age 66 years; 10 women) scheduled for primary unilateral total knee arthroplasty. The patients were treated on two days (day 7 and day 10) postoperatively. On one day they received 30 minutes of knee icing (active treatment) and on the other day they received 30 minutes of elbow icing (control treatment). The order of treatments was randomized. Maximal knee extension strength (primary outcome), knee pain at rest and knee pain during the maximal knee extensions were measured 2-5 minutes before and 2-5 minutes after both treatments by an assessor blinded for active or control treatment. The change in knee extension strength associated with knee icing was not significantly different from that of elbow icing (knee icing change (mean (1 SD)) -0.01 (0.07) Nm/kg, elbow icing change -0.02 (0.07) Nm/kg, P = 0.493). Likewise, the changes in knee pain at rest (P = 0.475), or knee pain during the knee extension strength measurements (P = 0.422) were not different between treatments. In contrast to observations in experimental knee effusion models and inflamed knee joints, knee joint icing for 30 minutes shortly after total knee arthroplasty had no acute effect on knee extension strength or knee pain.


Butein inhibits cancer cell growth by rescuing the wild-type thermal stability of mutant p53.
p53 is a transcription factor that activates the expression of various genes involved in the maintenance of genomic stability, and more than 50% of cancers harbor inactivating p53 mutations, which are indicative of highly aggressive cancer and poor prognosis. Pharmacological targeting of mutant p53 to restore the wild-type p53 tumor-suppressing function is a promising strategy for cancer therapy. In this study, we identified a small molecule, Butein, that reactivates mutant p53 activity in tumor cells harboring the R175H or R273H mutation. Butein restored wild-type-like conformation and DNA-binding ability in HT29 and SK-BR-3 cells harboring mutant p53-R175H and mutant p53-R273H, respectively. Moreover, Butein enabled the transactivation of p53 target genes and decreased the interactions of Hsp90 with mutant p53-R175H and mutant p53-R273H proteins, while Hsp90 overexpression reversed targeted p53 gene activation. In addition, Butein induced thermal stabilization of wild-type p53, mutant p53-R273H and mutant p53-R175H, as determined via CETSA. From docking study, we further proved that Butein binding to p53 stabilized the DNA-binding loop-sheet-helix motif of mutant p53-R175H and regulated its DNA-binding activity via an allosteric mechanism, conferring wild-type-like the DNA-binding activity of mutant p53. Collectively, the data suggest that Butein is a potential antitumor agent that restores p53 function in cancers harboring mutant p53-R273H or mutant p53-R175H. SIGNIFICANCE: Butein restores the ability of mutant p53 to bind DNA by reversing its transition to the Loop3 (L3) state, endows p53 mutants with thermal stability and re-establishes their transcriptional activity to induce cancer cell death.


Mycosinthetized Ag, CuO and ZnO nanoparticles from a promising Trichoderma harzianum strain and their antifungal potential against important phytopathogens.
Fungal green biosynthesis of nanoparticles (NPs) is a promising eco-friendly method for mass-scale production. In the present study Ag, CuO and ZnO nanoparticles were biogenically synthetized using a cell filtrate of a strain of Trichoderma harzianum as a reducer and stabilizer agent. The structure, morphology and physicochemical properties of the NPs were characterized through transmission electron microscopy, dynamic light scattering, wide angle X-ray scattering and thermogravimetric analysis. Since nanotechnology could offer promising applications in agricultural area, we evaluated the ability of the NPs to reduce the growth of important fungal phytopathogens as Alternaria alternata, Pyricularia oryzae and Sclerotinia sclerotiorum. Silver and CuO NPs reduced significantly the mycelial growth of A. alternata and P. oryzae in a dose dependent manner. This is the first report of a multiple extracellular biosynthesis of NPs from T. harzianum and the first time that CuO and ZnO NPs were obtained from this fungus. In addition, we highlighted the rapid production of NPs, as well as, the potential of Ag and CuO for the control of phytopathogens. On the other hand, the three types of NPs could be easily and sustainably produced on a large scale with the chance of having multiple applications in biotechnological processes.


Variability of Blood Eosinophil Count at Stable-State in Predicting Exacerbation Risk of Chronic Obstructive Pulmonary Disease.
Chronic obstructive pulmonary disease (COPD) phenotyping using stable-state blood eosinophil level was shown to have prognostic implication in terms of exacerbation risk. However, using a single cut-off of blood eosinophil level to predict clinical outcome has been challenged. There have been suggestions that variability of blood eosinophil count at stable-state could provide additional information on exacerbation risk. A retrospective cohort study was conducted in a major regional hospital and a tertiary respiratory referral centre in Hong Kong, including 275 Chinese patients with COPD, to investigate the possible role of variability of blood eosinophil count at stable-state to predict COPD exacerbation risk in one year. Higher variability of baseline eosinophil count, which is defined as the difference of the minimal and maximal eosinophil count at stable-state, was associated with increased risk of COPD exacerbation in the follow-up period with adjusted OR (aOR) of 1.001 (95% CI = 1.000-1.003, p-value = 0.050) for 1 unit (cells/µL) increase in variability of baseline eosinophil count, aOR of 1.72 (95% CI = 1.00-3.58, p-value = 0.050) for 1 SD increase in variability of baseline eosinophil count and aOR of 1.06 (95% CI = 1.00-1.13) for 50 cells/µL increase in variability of baseline eosinophil count. The AUC by ROC analysis was 0.862 (95% CI = 0.817-0.907, p-value < 0.001). The cut-off for variability of baseline eosinophil count identified was 50 cells/µL, with sensitivity of 82.9% and specificity of 79.3%. Similar findings were also shown in the subgroup with stable-state baseline eosinophil count below 300 cells/µL. Variability of baseline eosinophil count at stable-state might predict the exacerbation risk of COPD, exclusively among patients with baseline eosinophil count below 300 cells/µL. The cut-off value for variability was 50 cells/µValidation of the study findings in large scale prospective study would be meaningful.


Influence of cortisol on insulin- and insulin-like growth factor 1 (IGF-1)-induced steroid production and on IGF-1 receptors in cultured bovine granulosa cells and thecal cells.
During stress, hyperactivity of the adrenal gland can directly and indirectly inhibit ovarian function. However, little evidence existed to support the notion that glucocorticoids could influence insulin-like growth factor 1 (IGF-1) action within the ovary. Therefore, the effect of cortisol on IGF-1-induced granulosa and thecal cell function was evaluated. Granulosa and thecal cells from bovine ovarian follicles were cultured for 2 d in the presence of 10% fetal calf serum and then cultured for an additional 2 d in serum-free medium with added hormones. Cortisol had little or no effect (p > 0.05) on IGF-1-induced progesterone production by granulosa cells from both small (1-5 mm) or large (> or =8 mm) follicles. Also, cortisol had little or no effect (p > 0.05) on basal, insulin-, or IGF-1-induced estradiol production by granulosa cells from small or large follicles, or on the number of IGF-1 receptors in granulosa cells from small follicles. Cortisol had no effect (p > 0.10) on insulin-induced granulosa cell numbers, but increased IGF-1-induced granulosa cell numbers. In thecal cells, doses of 1-100 ng/mL of cortisol increased (p< 0.05) insulin- and IGF-1-induced thecal cell numbers by 10-20%, progesterone production by 18-36%, and androstenedione production by two- to fourfold. The estimated dose of cortisol necessary to stimulate 50% of the maximum androstenedione production in the presence of IGF-1 was 7 ng/mL. In contrast, cortisol decreased (p < 0.05) the number of IGF-1 receptors in thecal cells by 45%. In conclusion, cortisol at physiological levels can directly influence ovarian follicular function in cattle, especially thecal androstenedione production.


Preparation of Highly Biocompatible ZnSe Quantum Dots Using a New Source of Acetyl Cysteine as Capping Agent.
In this paper, we describe a facile method for preparation of ZnSe quantum dots (QDs) using an inexpensive and biocompatible source of acetyl cysteine in aqueous media. The structural properties of the ZnSe QDs have been characterized using XRD, FT-IR, and TEM techniques. The optical properties of the as-prepared QDs were found to be size-dependent, due to the strong confinement regime at relatively low refluxing time. Effect of solution pH and refluxing temperature on absorption and emission characteristics of the ZnSe QDs was studied. The empirical effective mass approximation also reveals that, both solution pH and refluxing temperature parameters would effect on ZnSe QDs growth, and increase their size. However, the influence of the solution pH was found to be more prominent. Water-solubility, high emission intensity and sub-10 nm nanocrystals size are the most essential features that suggest our synthesized aqueous-based ZnSe QDs (with a very cost-effective and biocompatible capping agent) can be utilized for biological intentions.


The Haemophilus influenzae Hap serine protease promotes adherence and microcolony formation, potentiated by a soluble host protein.
Haemophilus influenzae initiates infection by colonizing the upper respiratory mucosa. The process of colonization involves adherence to epithelium and evasion of host immunity. In this study, we examined the H. influenzae Hap adhesin, which has serine protease activity and undergoes autoproteolytic cleavage and extracellular release in broth. We found that the uncleaved cell-associated form of Hap mediates adherence to cultured epithelial cells and promotes bacterial aggregation and microcolony formation. Adherence and aggregation are augmented by secretory leukocyte protease inhibitor, a natural component of respiratory secretions that inhibits Hap autoproteolysis. These observations suggest a novel paradigm in host-pathogen relations, in which a soluble host protein whose primary function is to protect host epithelium potentiates properties that facilitate bacterial colonization.


Deciphering the induction of Listeria monocytogenes into sublethal injury using fluorescence microscopy and RT-qPCR.
The adaptive response of bacterial cells to changing environmental conditions depends on the behavior of single cells within the population. Exposure of Listeria monocytogenes to sublethal acidic conditions in foods or in the gastrointestinal track of the host may induce injuries relevant to difficult physiological states within the dormancy continuum. In this study, exposure to acidic conditions (acetic-AA and hydrochloric acid-HCl adjusted to pH 3.0, 2.7, 2.5 at 20 °C for 5 h) was used to evaluate injury of L. monocytogenes, Scott A strain. To differentiate the resistant sub-population from the total, Tryptic Soy Agar with 0.6 % Yeast Extract (TSAYE) supplemented or not with 5 % NaCl were comparatively used. Sublethally injured cells were detected by comparing plate counts with fluorescence microscopy, using combinations of CFDA (viability) and Propidium-Iodide (death). Effect of acid stress on the relative transcription of clpP, mazE, mazF, relA, gadC, gadD, gadB, sigB, inlA and prfA upon transition of total population into different physiological stages was evaluated through RT-qPCR. AA treated cells showed measurable logarithmic reduction at pH 2.7 and 2.5, while there was a significant percentage of CFDA-/PI+ cells. Evaluation of the potentially culturable population on TSAYE, from the percentage of CFDA/PI-stained cells, revealed that unstained cells represented a non-culturable sub-population. Exposure to Ringer's solution pH 2.7, adjusted with AA, resulted in higher percentages of non-esterase active with membrane integrity cells (CFDA-/PI-) compared to the percentages of the enumerated culturable cells on TSAYE after 4 and 5 h. Under the same conditions, after 1 h of exposure macroscopic observation revealed size colony variations (SCVs) of the total population (CFU on TSAYE). L. monocytogenes retained its culturability after hydrochloric acid exposure, while cells remained metabolically active (CFDA+). However, a stochastic change in cell's shape, was detected after exposure to pH 3.0 and 2.5, adjusted with HCl, for 2 h at 20 °C. A pattern of gene up-regulation was observed during treatment with AA pH 2.7 and HCl pH 3.0 at the 3rd h of exposure. Deciphering L. monocytogenes sublethal injury sheds light into the physiological and molecular characteristics of this state and provides the food science community with quantitative data to improve risk assessment.


Liveborn children and risk of hepatocellular carcinoma.
Clinical, animal, and epidemiologic evidence indicates that exogenous steroids influence the risk of hepatocellular carcinoma (HCC) and a recent study suggested that parity also may increase the risk of this tumor in women. The latter hypothesis was evaluated in the data from a case-control study which was carried out in Athens and covered 166 male and 19 female cases of HCC, and 381 male and 51 female hospital controls. Among males, there was no association between the number of liveborn children and risk of HCC, whereas among women, there was a suggestive positive association. Compared with women with one or two children, the relative risk for HCC was 0.6 among nulliparous women, 1.3 among those with three or four children and 1.7 among those with five or more children. The association of parity with risk of HCC was limited to women who were positive for hepatitis-B surface antigen (HBsAg) and was not confounded by hepatitis-C virus infection or tobacco smoking. The small number of HCC cases does not permit firm conclusions. If confirmed, however, these results would provide the foundation for a practical preventive advice that could be given to women who are positive for HBsAg.


Self-directed physical activity interventions for motor symptoms and quality of life in early and mid-stage Parkinson's disease: A systematic review and meta-analysis.
Self-directed physical activity (SDPA) has been found in previous research to alleviate some of the symptoms of early and mid-stage Parkinson's disease (PD) patients. So, this study aimed to determine the impact of SDPA on these patients in the areas of motor symptoms, and quality of life (QoL). PubMed, Embase, the Cochrane Library, the Web of Science, Scopus, and ProQuest were all searched. The risk-of-bias tool of Cochrane for randomized trials, Version 2 (RoB 2), was also used to assess the credibility of studies in terms of their procedures. 15 eligible studies were analyzed. SDPA improved motor function and mobility, including timed up and go (TUG) test [standardized mean difference (SMD), -0.55 (95% CI, -0.91 - -0.18), p = 0.003], 6-minute walking (6MW) test [1.11 (0.75 - 1.47), p < 0.00001], stride length [0.45 (0.18 - 0.72), p = 0.001], gait velocity [0.42 (0.04 - 0.81 p = 0.03], Unified Parkinson's Disease Rating Scale Part-III (UPDRS-III) [-0.76 (-1.18 - -0.33), p = 0.0005] and enhanced Berg Balance Scale (BBS) [0.88 (0.50 - 1.27), p < 0.00001]). Despite engaging in SDPA, there was no significant improvement observed in freezing of gait (FOG) [0.23 (-0.11 - 0.56), p = 0.18] as well as the Parkinson's Disease Questionnaire-39 (PDQ-39) [0.04 (-0.55 - 0.62), p = 0.90]). The motor symptoms of those with early to mid-stage PD improved with SDPA, however the research found no enhancement in FOG or QoL.


Retrospective Analysis of Fondaparinux and Low-Molecular-Weight Heparin in the Treatment of Women With Recurrent Spontaneous Abortion.
To compare the clinical efficacy of fondaparinux and LMWH and provide clinical evidence for the effectiveness of fondaparinux in the treatment of recurrent spontaneous abortion caused by PTS. A retrospective analysis was conducted for 120 patients diagnosed with a recurrent spontaneous abortion caused by PTS in Qingdao Jinhua Women's Hospital from March 2019 to April 2020. The patients were divided into two groups: 68 cases in the control group, treated with LMWH, 52 cases in the observational group, treated with fondaparinux. The pregnancy outcomes and adverse reactions between the two groups of recurrent miscarriage patients were compared. No significant difference was detected in the general data between the two groups of patients before treatment (P>0.05). In the observational group, the R value was increased, and the α and MA values were decreased after three months of treatment compared to those before treatment (P<0.05). In the control group, the R value was increased, and the MA value was decreased after three months of treatment compared to those before treatment (P<0.05). After treatment, no significant difference was observed in the pregnancy outcome between the two groups (P>0.05). The total adverse reaction rate of the fondaparinux group was lower than that of the LMWH group (P<0.05). In this study, no significant difference was detected in the pregnancy outcome between fondaparinux and LMWH in the treatment of recurrent spontaneous abortion caused by PTS, but fondaparinux had a low occurrence rate of adverse reactions and high safety.


Heterologous expression of Homo sapiens alpha-folate receptors in E. coli by fusion with a trigger factor for enhanced solubilization.
The role of Alpha folate receptors (FRα) in folate metabolism and cancer development has been extensively studied. The reason for this is not only associated to its direct relation to disease development but also to its potential use as a highly sensitive and specific biomarker for cancers therapies. Over the recent years, the crystal structures of human FRα complexed with different ligands were described relying on an expensive and time-consuming production process. Here, we constructed an efficient system for the expression and purification of a human FRα in E. coli. Unlike a conventional expression method we used a specific protein fusion expressing the target protein together with a trigger factor (TF). This factor is a chaperone from E. coli that assists the correct folding of newly synthesized polypeptide chains. The activity of rTFFRα was comparable to glycosylphosphatidylinositol (GPI) anchored proteins extracted from HeLa tumor cells. Our work demonstrates a straightforward and versatile approach for the production of active human FRα by heterologous expression; this approach further enhances the development of inhibition studies and biotechnological applications. The purified product was then conjugated to liposomes, obtaining a 35% higher signal from densitometry measurement on the immunoblotting assay in the contruct containing the Ni-NTA tag, as a mimesis of an exosome, which is of vital importance to nanotherapeutic techniques associated to treatment and diagnosis of tumors.


Expression of an Aspergillus niger xylanase in yeast: Application in breadmaking and in vitro digestion.
The cDNA of the β-1,4-endoxylanase of Aspergillus niger US368 was cloned and expressed in Pichia pastoris under the constitutive GAP promoter. The maximum activity obtained was 41 U mL(-1), which was about 3-fold higher than that obtained with the native species. The purified enzyme showed a specific activity of 910 U mg(-1) and a molecular mass of 24 kDa. It had an optimal activity at pH 4 and 50 °C, stable in a wide range of pH and in the presence of some detergents and organic solvents. r-XAn11-His6 (recombinant xylanase) was used as an additive in breadmaking. A decrease in water absorption, an increase in dough rising and improvements in volume and specific volume of the bread were recorded. The r-XAn11-His6 was also used in in vitro digestion of barley and wheat bran leading to a decrease of the viscosities and an increase of the reducing sugars and total sugars contents.


A Nomogram Based on the Log Odds of Positive Lymph Nodes Predicts the Prognosis of Patients with Colon Neuroendocrine Tumors After Surgery: A Surveillance, Epidemiology, and End Results Population-Based Study.
This work focused on determining the highly efficient nodal classification system from American Joint Committee on Cancer (AJCC) tumor node metastasis (TNM) classification (eighth edition), positive lymph node, log odds of positive lymph nodes (LODDS), lymph node ratio, examined lymph node, and establishing the new nomogram for predicting cancer-specific survival in colon neuroendocrine tumors (CNETs). From the Surveillance, Epidemiology, and End Results database, 943 CNET cases undergoing radical operation were enrolled, and randomized as training (n = 663) or validation set (n = 280). For the above 5 lymph node classification systems, their prediction performances were compared with C-index, Akaike information criterion (AIC), and area under the receiver operating characteristic curve. Univariate together with multivariate regression was carried out for identifying independent risk factors. Afterward, this work established 1 nomogram and confirmed its accuracy based on C-index, calibration curves, together with the area under the curve value. Besides, it was compared with the AJCC TNM classification system with regard to model prediction performance. LODSS achieved the greatest area under the curve and C-index, whereas the smallest AIC. Upon multivariate regression, age, histologic grade, T stage, M stage, and LODDS independently predicted the risk of CNETs. For the validation set, the C-index of the nomogram was 0.794, and the area under the curves at 1, 3, and 5 years was 0.826, 0.857, and 0.870, separately. Additionally, as revealed by the C-index, AIC, decision curve analysis, as well as Kaplan-Meier analysis, our nomogram had superior performance to the AJCC TNM classification system. For postoperative patients with CNETs, the LODDS might achieve the best prediction performance. Moreover, the LODDS-based nomograms might show superior survival prediction performance to the AJCC TNM classification system (eighth edition).


[In vitro analysis of diffusion through radicular tissue of paraformaldehyde contained in a paste commonly used in endodontics].
The aim of the study was to show how paraformaldehyde contained in a preparation normally used in endodontic therapy spreads through the root tissues. The results reveal that paraformaldehyde has the same particular tendency as formaldehyde to spread in the periapex, maximum levels are reached after 48 hours and tend to decrease after 72 and 96 hours.


Safe and effective topical application dose of lidocaine for surgery with laryngomicroscopy.
A clinical case of a patient in whom seizure developed after topical application of lidocaine to the oropharyngeal region was described. The exploratory studies on the optimal dosage of lidocaine in the surgery under laryngomicroscope was performed in 22 patients. During the application of 2% lidocaine viscous and 4% lidocaine solution to the oropharyngeal region, the patient was instructed to expectorate the excess intermittently to avoid absorption. In addition, the oropharyngeal region was swabbed with gauze. The serum lidocaine concentrations after application were less than 1.8 micrograms/ml. The total application dose of the drug was within the range of 324 to 640 mg, and the total recovery rate from the saliva and gauze was 52% to 81%. We found a good correlation between net application dose (total application dose minus excessive lidocaine dose) and serum lidocaine concentration. In this study, it was shown that the safe and effective net application dose of lidocaine for the surgery may be within the range of 127 to 260 mg.


Study of the automated breast tumor extraction using 3D ultrasound imaging: The usefulness of depth-width ratio and surface-volume index.
We applied quantitative parameters in three-dimensional ultrasonic images to distinguish benign from malignant breast tumors in 29 benign cases including 8 cysts and 21 fibroadenomas, and 32 malignant cases including 23 ductal carcinomas, 2 special types of carcinoma, 1 malignant lymphoma and 6 others. This procedure involved simultaneously acquiring video data from real-time ultrasonic images and recording the original position and orientation of the probe. Both sets of data were fed directly into a desktop computer. Fuzzy reasoning and relaxation techniques were use to automatedly extract the shape of the tumor and render it in three dimensions. We then evaluated three parameters: 2D-D/W, the so-called depth-width ratio measured in B-mode images: 3D-D/W; and the S/V index ([surface area](3)/36π [volume](2)) calculated from the three-dimensional volume extracted with this system. All three parameters were significantly higher in the malignant group (averages: 0.81, 0.64, and 11.3, respectively) than in the benign group (averages: 0.62, 0.47, and 3.78, respectively). All three parameters were thus found to be useful in differentiating the two groups.


Di-μ2-chlorido-bis-{chlorido-[2,4,6-tris-(pyridin-2-yl)-1,3,5-triazine-κ3 N 2,N 1,N 6]nickel(II)}.
In the title compound, [Ni2Cl4(C18H12N6)2], the NiII ions are hexa-coordinated in a distorted octa-hedral coordination environment defined by three N atoms of the tridentate 2,4,6-tri-2-pyridyl-1,3,5-triazine ligand and three Cl- anions in a meridional geometry. The two NiII ions are bridged by two Cl anionic ligands, thereby forming a dinuclear complex. A crystallographic centre of inversion is located at the centroid of the Ni2Cl2 ring.


Integrated vector management program in the framework of the COVID-19 pandemic in Medellin, Colombia
Introduction: The COVID-19 pandemic pressured health care systems to remain alert and active in their vector-borne disease control and prevention programs, leading to changes in vector control strategies in urban areas affected by dengue, Zika and chikungunya. Objective: To describe the adaptations made to the vector control and surveillance program in Medellín during the COVID-19 health emergency. Materials and methods: Once the health emergency started, biosecurity protocols were developed. Entomological surveillance was strengthened from the institutional environment instead of homes. Data was collected in Medellín from 2018 to 2021 during the vector control and surveillance program activities, which included epidemiological and entomovirological surveillance, entomological index survey, ovitrap monitoring, community mobilization, search and elimination of mosquito breading sites, and chemical control. These actions were adapted and/or increased to promote self-care among communities in total and partial confinement, and to develop prevention and control measures. Results: Mosquito monitoring was increased by 40% using ovitraps, entomological virological surveillance showed an increase in 2020 of 34,4% compared to 2019 and virtual media was used to keep and improve contact with the community. Conclusion: The COVID-19 pandemic had a significant impact on arbovirus prevention and control programs. The city of Medellín quickly adapted its entomo-virological surveillance activities, control measures, and the contact with the community during the pandemic, which allow the Integrated Vector Management program to remain active in the city.


Identification of an intestinal neutral glycosphingolipid as a phenotype-specific receptor for the K88ad fimbrial adhesin of Escherichia coli.
In this study, we identified a receptor for the K88ad fimbrial adhesin of Escherichia coli in neutral glycosphingolipid preparations from intestinal epithelial cells of K88ad-adhesive pigs, which was absent in preparations from K88ad-nonadhesive pigs. Neither K88ab nor K88ac adhesin variants bound to this neutral glycosphingolipid. Because this receptor is an intestinal glycosphingolipid that binds K88ad adhesin, it has been designated IGLad. Carbohydrate compositional analysis of a partially purified preparation of IGLad identified galactose, glucose, and N-acetylglucosamine in a ratio of 1.5:1.0:0.5 as the major monosaccharides. Preliminary characterization experiments using lectins showed that IGLad contains the terminal glycanic structure Galbeta1-4GlcNAc. Removal of terminal beta-linked galactose residues from IGLad decreased the recognition of IGLad by the K88ad adhesin, indicating that terminal beta-linked galactose is an essential component of the K88ad adhesin recognition site on IGLad. Studies with purified glycosphingolipid standards demonstrated that K88ad adhesin binds to neolactotetraosylceramide (nLc4Cer) (Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer) , lactotriosylceramide (GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer) and lactotetraosylceramide (Galbeta1-3GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer) . Based on these studies, IGLad appears to be nLc4Cer.


Kill 'Em All: Efgartigimod Immunotherapy for Autoimmune Diseases.
Neonatal Fc receptors (FcRns) recycle IgGs by preventing their lysosome degradation. As this process also enhances half-life of pathogenic auto-IgG, inspired from the mechanisms of intravenous immunoglobulin, several inhibitors of IgG-FcRn interface have been conceived for treating autoimmune diseases. Among them, the high-affinity FcRn-binding engineered Fc molecule efgartigimod has recently completed phase I clinical trial.


Olfactory Dysfunction and Balance Dysfunction are Associated with Increased Falls in Older Adults.
This study aims to characterize the association between impairments in olfaction and balance, both of which are mediated in part by the cerebellum, and how this relates to prospective incidence of falls in a cohort of aging adults. The Health ABC study was queried to identify 296 participants with data on both olfaction (measured using the 12-item Brief Smell Identification Test) and balance-related function (measured using the Romberg test). The relationship between olfaction and balance was investigated using multivariable logistic regression. Predictors of performance on a standing balance assessment and predictors of falls were studied. Of 296 participants, 52.7% had isolated olfactory dysfunction, 7.4% had isolated balance dysfunction, and 5.7% had dual dysfunction. Severe olfactory dysfunction was associated with increased odds of balance dysfunction when compared to those without olfactory dysfunction, even when adjusting for age, gender, race, education, BMI, smoking, diabetes, depression, and dementia (OR = 4.1, 95% CI [1.5, 13.7], p = 0.011). Dual sensory dysfunction was associated with worse performance on a standing balance assessment (β = -22.8, 95% CI [-35.6, -10.1], p = 0.0005) and increased falls (β = 1.5, 95% CI [1.0, 2.3], p = 0.037). This study highlights a unique relationship between olfaction and balance, and how dual dysfunction is associated with increased falls. With substantial implications of falls on morbidity and mortality in older adults, this novel relationship between olfaction and balance emphasizes a potentially shared mechanism between olfactory dysfunction and increased fall risk in older adults; however, further study is required to explore the novel relationship of olfaction with balance and future falls. 3 Laryngoscope, 133:1964-1969, 2023.


Use of exposure time and life expectancy in models for toxicity to aquatic organisms.
The exposure time is a variable which is usually not incorporated into models for toxicity. However, with persistent organic pollutants (POPs) exhibiting a nonspecific mode of action and lipophilic properties this variable can be modeled by the usage of the internal concentrations as a measure of the toxicity with fish. The bioconcentration process with fish is a relatively well understood and predictable process which allows the calculation of the internal lethal concentration. When the exposure time is relatively short the critical internal lethal concentrations are relatively constant for the group of POPs whereaas the LC50 measured in the ambient water is quite variable. When the exposure time is relatively long, results on the measurement of the critical internal concentration with fish over different exposure times has demonstrated that the internal lethal concentration falls with increasing exposure times in a consistent and predictable manner. This reduction in life expectancy can be described in a model which can be used to estimate the critical internal concentration for any exposure time. It also provides information useful in assessing the risk to fish and potentially other species due to the occurrence of residues of POPs in natural aquatic systems. It is suggested that these relationships can be extended to other groups of organisms and chemicals.


Similarities between G-proteins in visual cells of Sepia and cattle.
In contrast to antisera against native transducin a polyclonal antiserum raised against heat-denatured bovine transducin crossreacts with the G-protein from Sepia visual cells. This antiserum recognizes a 44 kDa (G alpha) and a 36 kDa (G beta) protein band from Sepia photosensory membrane preparation. Furthermore we purified the antibody-binding G-protein from Sepia by binding it to light-activated rhodopsin of Sepia and GTP-induced extraction, similar to the purification of bovine transducin. This G-protein is probably involved in the phototransduction process. The purified Sepia G-protein did bind to vertebrate photosensoric membrane upon illumination, but was not eluted by GTP-containing buffer solution. After extensive bleaching, the G-protein became soluble.


Cathepsin B of Cynoglossus semilaevis: identification, expression, and activity analysis.
Cathepsin B (EC 188.8.131.52) is a member of the papain family cysteine protease and in mammals is known to be involved in protein degradation and other biological functions. However, very little is known about the function of cathepsin B in fish. In this study, we identified and analyzed a cathepsin B homologue (CsCatB) from tongue sole (Cynoglossus semilaevis, Pleuronectiformes), an economic fish species cultured in China. CsCatB is composed of 322 amino acid residues and shares 70-81.3% overall sequence identities with its counterpart in teleosts and humans. CsCatB possesses typical cathepsin B structural features including the propeptide region and the papain family cysteine protease domain, the latter containing the four catalytic residues (Q101, C107, H277, and N297) that are conserved in lower and higher vertebrates. Quantitative real time RT-PCR analysis showed that CsCatB expression occurred in multiple tissues and was positively regulated by bacterial infection and by immunization with a subunit vaccine. Recombinant CsCatB purified from Escherichia coli exhibited apparent protease activity, which was optimal at 35 °C and pH 5.5. In contrast, a mutant CsCatB bearing glutamic acid substitution at H277 was dramatically reduced in proteolytic activity. These results indicate that CsCatB is a biologically active protease that is likely to be involved in host immune response during bacterial infection and vaccination.


Silencing of miR-101 Prevents Cartilage Degradation by Regulating Extracellular Matrix-related Genes in a Rat Model of Osteoarthritis.
Osteoarthritis (OA) is a common, degenerative joint disease characterized by articular cartilage degradation. Currently, clinical trials based on microRNA therapy have been performed to treat various diseases. However, no treatment has been found for arthritis. This study investigated the functions of miR-101 in cartilage degradation in vivo and evaluated the feasibility of using miR-101 as a therapeutic agent for OA. Mono-iodoacetate-induced arthritis (MIA) rats were used as an animal model of OA. miR-101 mimic or miR-101 inhibitor was injected into the rats' knees to evaluate its effects on cartilage degradation. Cartilage degradation aggravated at 14 days after the injection of miR-101 mimic. By contrast, miR-101 silencing reduced cartilage degradation. Moreover, the administration of miR-101 mimic is sufficient to cause cartilage degradation in the normal cartilage of rats. By contrast, miR-101 inhibitor could prevent this change. Microarray and qPCR were used to investigate the different expressed genes after injecting miR-101 mimic and miR-101 inhibitor in the rats' articular cartilage. Several cartilage degradation-related genes were selected and validated to function in cartilage degradation with miR-101. Our results demonstrated the therapeutic effect of miR-101 inhibition on cartilage degradation in MIA rats by regulating several cartilage degradation-related genes.


Injectable and biocompatible alginate-derived porous hydrogels cross-linked by IEDDA click chemistry for reduction-responsive drug release application.
In this work, novel injectable and reduction-responsive hydrogels were successfully prepared via inverse electron demand Diels-Alder reaction between alginate-norbornene and a water-soluble PEG based disulfide cross-linker. The reduction-responsive cross-linker was designed to contain a PEG chain within two disulfide linkages, and two terminal tetrazine groups. The resulting hydrogels possessed high swelling ratios, porous morphology, excellent drug loading efficiency (~92%), and suitable mechanical properties. The drug release experiments demonstrated that the hydrogels released more than 90% of the encapsulated doxorubicin (DOX) in the presence of 10 mM glutathione while a minimal DOX release (<25%) was measured in physiological buffer (PBS, pH = 7.4) after 11 d. The cross-linker and hydrogels did not exhibit any apparent cytotoxicity to fibroblast cells. In contrast, DOX-loaded hydrogels induced anti-tumor activity against cancer cells. The injectable and reduction-responsive hydrogels hold great potential as a biomaterial for stimuli responsive drug delivery applications.


Recent progress in self-powered multifunctional e-skin for advanced applications.
Electronic skin (e-skin), new generation of flexible wearable electronic devices, has characteristics including flexibility, thinness, biocompatibility with broad application prospects, and a crucial place in future wearable electronics. With the increasing demand for wearable sensor systems, the realization of multifunctional e-skin with low power consumption or even autonomous energy is urgently needed. The latest progress of multifunctional self-powered e-skin for applications in physiological health, human-machine interaction (HMI), virtual reality (VR), and artificial intelligence (AI) is presented here. Various energy conversion effects for the driving energy problem of multifunctional e-skin are summarized. An overview of various types of self-powered e-skins, including single-effect e-skins and multifunctional coupling-effects e-skin systems is provided, where the aspects of material preparation, device assembly, and output signal analysis of the self-powered multifunctional e-skin are described. In the end, the existing problems and prospects in this field are also discussed.


Ageing reduces the reproductive performance of male white shrimp Litopenaeus vannamei by altering sperm intracellular Ca2+ concentrations and interfering with sperm apoptosis.
Although the effects of age on the reproductive performance of various invertebrates, including white shrimp Litopenaeus vannamei are increasingly well documented, the mechanisms manifesting these impacts remain poorly understood. To ascertain the mechanisms of age on reproductive performance, the sperm quality, intracellular contents of Ca2+, insemination and hatching rates, and status of sperm apoptosis in terms of the expression of key regulatory genes were investigated in 11 and 16 month old male L. vannamei. The aged male individuals (16 months) had lesser reproductive performance in terms of fertilization and hatching rates. In addition, fewer and less viable sperm were detected in aged shrimp, which may be due to the altered expression of apoptosis-related genes. Furthermore, the aged males had lesser intracellular contents of Ca2+ in the sperm which may decrease the capacity of these gametes to undergo a complete acrosome reaction. In general, due to the decrease in intracellular contents of Ca2+ and alterations in the process of apoptosis, aged L. vannamei have relatively lesser quality sperm and may, therefore, have lesser reproductive performance.


Maturation of the auditory brainstem in low risk-preterm infants: a comparison with age-matched full term infants up to 6 years.
Whether preterm birth has a significant effect on the maturation of the human brain remains an equivocal issue. Since experience plays a crucial role in the development of the brain, it is conceivable that extra-uterine preterm exposure could exert some effects on the maturation of the developing human brain. The present study compared the post-term maturation of the central components of brainstem auditory evoked response (BAER) in low risk preterm infants with that of age-matched full term infants up to 6 years. The preterm infants demonstrated similar maturational profiles to those of the term infants in both interpeak intervals and amplitude measures. No systematic, statistically significant differences were found between the preterm and term infants in any of the BAER variables although the I-V interval tended to be slightly shortened in the prematurely born infants. These results suggest that preterm birth or earlier exposure to sound environment extra utero is unlikely to lead to significant neurophysiological consequence in the developing auditory brainstem of low risk infants.


Intragraft drug infusion as an adjunct to balloon catheter thrombectomy for salvage of thrombosed infragenicular vein grafts: a preliminary report.
Early infragenicular vein graft thrombosis is associated with poor secondary patency, particularly when no correctable defect is identified. We have attempted to improve patency of thrombosed vein grafts by direct infusion of vasodilator and anticoagulant drugs after surgical thrombectomy. Among 212 infragenicular vein grafts, 16 (7.5%) required thrombectomy within 30 days of surgery (14 in situ saphenous vein, 1 composite vein, and 1 cephalic vein graft). Causes for failure were corrected in four (graft twist, intimal tear, suture failure, and external compression), resulting in prolonged patency. No cause for failure was apparent in the 12 remaining grafts after balloon catheter thrombectomy and arteriography. Two of these grafts occluded within 10 days despite multiple attempts at vein patch angioplasty, distal graft extension, and repeat thrombectomy with systemic anticoagulation. In the remaining 10 grafts, a small polyethylene catheter was placed in a proximal vein branch for direct intragraft drug infusion. Heparin (10 units/min) and nitroglycerin (50 micrograms/min) were the agents infused most frequently, for a mean duration of 52 hours after thrombectomy. Of these 10 infused grafts, 8 remained patent during a mean 17-month follow-up (range, 6 to 38 months). This was accomplished despite previous and repeated failures of thrombectomy and systemic anticoagulation in seven of these eight grafts. Two infused grafts rethrombosed within 30 days of infusion, resulting in amputation. No catheter-related complications occurred. Increased thrombogenicity, intimal injury, and spasm after balloon catheter thrombectomy may contribute to vein graft rethrombosis in the absence of technical defects. Direct intragraft infusion of nitroglycerin and heparin contributed to prolonged salvage of 80% of thrombosed vein grafts in this preliminary experience.


Gastroschisis reduction using "Applied Alexis", a wound protector and retractor.
We used an Applied Alexis (Applied Medical Resources Corp, USA) wound protector and retractor (WPAR) instead of a "spring loaded silo" (SLS) for gastroschisis reduction because SLS is currently unavailable in Japan. Our patient was a 1,707 g female diagnosed prenatally with gastroschisis born at 35 weeks gestation by spontaneous vaginal delivery. Attempted primary closure failed and a WPAR was used as a silo. A piece of the bottom ring of the WPAR was cut out and a smaller ring was made from it, so it would fit into the patient's abdomen. Eviscerated organs were inserted into the retractor through the bottom ring, and the bottom ring was placed into the abdominal cavity through the gastroschisis defect without suturing to complete the silo. Operating time was 20 minutes. Three days later, all eviscerated organs had been reduced into the peritoneal cavity, the silo was removed, and the gastroschisis defect closed without difficulty. Postoperative recovery was uneventful and the WPAR was an excellent alternative for making a silo.


Monitoring a cohort of trainees: changes over time and associations between health literacy, health behaviour and health.
For many entrants, the initial stages of professional training are a challenge. Demands at work can lead to new physical and psychological stress, as well as new social requirements. These new requirements can influence the health behaviour and the state of health of young employees. In recent years, there have been many studies on health literacy (HL). HL represents resources and potentials that allow individuals to achieve improved control of their health and of factors that influence health. Thus, HL can influence both well-being and health. In the present study, the health of trainees in different branches (health and welfare services, office, sales, technology) is monitored over time ending in the period of the COVID-19 pandemic. Furthermore, the association between health literacy and health or health behaviour has been examined. In 2017/18, a baseline survey (T0) was performed on trainees in various sectors (office, sales, teaching, nursing and social welfare, engineering, hairdressers), who had been contacted through vocational colleges in four federal states in north Germany. The trainees were surveyed again in the in the first year after training in 2021 (T3). Demographic data were collected, as well as information on health literacy (HLS-EU-16), health behaviour (physical exercise, nutrition, smoking and alcohol) and state of health (BMI, psychological well-being and subjective state of health). Recognition, satisfaction at work and thoughts of leaving the profession were surveyed with the Copenhagen Psychosocial Questionnaire (COPSOQ). Statistical analysis was performed with tests for paired samples and multivariate regression analysis in SPSS 26. 129 data sets were evaluated, with a follow-up rate of 10.2%. 85% of the trainees were female. The mean age at follow-up was 25.6 years. 56% were employed in the health service or social welfare. 35% worked in the office, sales or engineering. At T3, 47% of the employees exhibited limited health literacy, 67% low levels of exercise and 30% risky alcohol use. 42% exhibited overweight and 42% poor psychological well-being. An association between health literacy (HL) and psychological well-being was only observed in cross-section (HL inadequate vs. HL adequate OR: 3.2 95% CI: 1.07-9.49, p = 0.037). The odds ratio relative to subjective state of health was also increased, although the association was not statistically significant (HL inadequate vs. HL adequate OR: 2.7 95% CI: 0.72-9.78, p = 0.143). In the sector for health service and social welfare, there was statistically significant deterioration over time in all COPSOQ variables (recognition, satisfaction at work, thoughts of leaving the profession). For a group of trainees in the first year of work during the covid-19 epidemic, the present findings show that there is a need to prevent risky health behaviour, overweight and poor psychological well-being. Health literacy was shown to be a modifiable parameter, that apparently is associated in cross - but not in longitudinal section with the health of young employees. It would appear to be reasonable to modify developing health literacy in the setting of work and school.


Antidepressant prescriptions, discontinuation, depression and perinatal outcomes, including breastfeeding: A population cohort analysis.
To explore associations between exposure to antidepressants, their discontinuation, depression [medicated or unmediated] and preterm birth [<37 and <32 weeks], small for gestational age (SGA) [<10th and <3rd centiles], breastfeeding [any] at 6-8 weeks. Design: A population-based cohort study. Setting: The Secure Anonymised Information Linkage [SAIL] databank in Wales, linking maternal primary care data with infant outcomes. Participants: 107,573, 105,331, and 38,725 infants born 2000-2010 with information on prematurity, SGA and breastfeeding respectively, after exclusions. Exposures: Maternal antidepressant prescriptions in trimesters 2 or 3, discontinuation after trimester 1, recorded diagnosis of depression [medicated or unmediated] in pregnancy. Methods: Odds ratios for adverse pregnancy outcomes were calculated, adjusted for smoking, parity, socio-economic status, and depression. Exclusive formula feeding at 6-8 weeks was associated with prescriptions in trimesters 2 or 3 for any antidepressants (adjusted odds ratio [aOR] 0.81, 95% confidence intervals 0.67-0.98), SSRIs [aOR 0.77, 0.62-0.95], particularly higher doses [aOR 0.45, 0.23-0.86], discontinuation of antidepressants or SSRIs after trimester 1 (aOR 0.70, 0.57-0.83 and 0.66, 0.51-0.87), diagnosis of depression aOR 0.76 [0.70-0.82), particularly if medicated (aOR 0.70, 0.58-0.85), rather than unmedicated (aOR 0.87, 0.82-0.92). Preterm birth at <37 and <32 weeks' gestation was associated with diagnosis of depression (aOR 1.27, 1.17-1.38, and 1.33, 1.09-1.62), particularly if medicated (aOR 1.56, 1.23-1.96, and 1.63, 0.94-2.84); birth at <37 weeks was associated with antidepressants, (aOR 1.24, 1.04-1.49]. SGA <3rd centile was associated with antidepressants (aOR 1.43, 1.07-1.90), and SSRIs (aOR 1.46, 1.06-2.00], particularly higher doses [aOR 2.10, 1.32-3.34]. All adverse outcomes were associated with socio-economic status and smoking. Exposure to antidepressants or depression increased risks of exclusive formula feeding at 6-8 weeks, and prescription of antidepressants was associated with SGA <3rd centile. Prescription of antidepressants offers a useful marker to target additional support and additional care before and during pregnancy and lactation.


Magnetic resonance imaging characteristics of thymoma in Vietnamese patients with myasthenia gravis in relation to histopathological type and disease staging.
Myasthenia gravis (MG) is a relatively uncommon neuromuscular disease. The thymus plays an important role in the pathogenesis of MG, with 70% of patients presenting with thymic follicular hyperplasia and 20% presenting with thymoma. This study aimed to characterize magnetic resonance (MR) images of thymomas in patients with MG, associated with various stages and types. A prospective study was performed in 46 MG patients with thymoma, who underwent surgery between August 2014 and September 2018, in Vietnam. Differences in MR imaging (MRI) images, according to type and stage, were assessed by the χ2 test or Fisher's exact test and Student's t-test. A p-value < 0.05 was considered significant. The proportions of each type of thymomas found to be invasive were 28.6%, 61.1%, 66.7%, 90.9%, and 100%, for type A, AB, B1, B2, and B3 thymomas, respectively. Heterogeneous signal intensities on T1- and T2-weighted images, necrosis, and cysts were more commonly observed in high-risk thymomas than in low-risk thymomas. Non-invasive thymomas were more likely to display smooth margins than invasive thymomas (p < 0.05). Only 16.7% of invasive thymomas were detected in MR findings. Only a few characteristic MRI features could be used to differentiate thymomas by stage or type.


Contemporary prostate cancer prevalence among T1c biopsy-referred men with a prostate-specific antigen level < or = 4.0 ng per milliliter.
To investigate the prostate cancer (PCa) prevalence and risk factors of men with prostate-specific antigen (PSA) level< or =4.0 ng/ml and an unsuspicious digital rectal examination (DRE) in a large biopsy referral cohort. Between 1997 and 2005, 855 men underwent initial transrectal ultrasound (TRUS)-guided prostate biopsy at the University Hospital Hamburg-Eppendorf. Patients with any previous surgical or medical treatment were excluded from analyses. Logistic regression analyses were performed to determine risk factors of PCa at biopsy and high-grade PCa defined as biopsy Gleason sum> or =7. Overall PCa detection rate was 23.1%. The majority had a biopsy Gleason sum of 6 (79.5%) and 20.5% had a biopsy Gleason sum> or =7. Total PSA (tPSA) and percentage of free PSA (%fPSA) were statistically significantly different in men with and without PCa (all p<0.001). In tPSA strata < or = 0.5, 0.6-1.0, 1.1-2.0, 2.1-3.0, and 3.1-4.0 ng/ml, PCa prevalence was 4.0%, 10.6%, 14.8%, 24.5%, and 32.1%, respectively. In logistic regression analyses addressing PCa and Gleason sum > or = 7 at biopsy, %fPSA and prostate volume represented independent and most informative risk factors. Our data demonstrate that a substantial percentage (23.1%) of men with a PSA< or =4.0 ng/ml and an unsuspicious DRE in a biopsy referral population harbor PCa, with 20.5% being high grade. Low %fPSA and low prostate volume represent important parameters in PCa and in high grade disease detection at biopsy, respectively.


Quality of Life and Behavioural Assessment in Post-adenotonsillectomy Cases of Paediatric Age Group.
Adenotonsillectomy is a widely practiced surgical intervention to manage obstructive sleep apnoea syndrome (OSAS) in the paediatric age group. We conducted a prospective, non randomised, interventional study among 40 patients in paediatric age group (2-18 years) suffering from adenotonsillar hypertrophy with an indication of adenotonsillectomy based on American Academy of Otolaryngology-Head and Neck Surgery criteria. Care givers completed the obstructive sleep apnoea 18 (OSA 18) quality of life (QoL) survey and Rutter Children's Behaviour Questionnaire (RCBQ) before adenotonsillectomy as well as 6 months after surgery. We found that mean score of OSA 18 and total RCBQ decreased significantly 6 months after surgery. The mean score of OSA 18 (4.12 ± 1.4) and total RCBQ score (20.5 ± 2) significantly reduced to 1.19 ± 0.12 and 7.4 ± 2.55 (p < 0.0001), respectively, after surgery. There was significant improvement in all the domains of OSA 18 score after intervention including sleep disturbance (mean score change 2.56, p < 0.0001), physical suffering (2.99, p < 0.0001), emotional distress (2.71, p < 0.0001), daytime problems (2.56, p < 0.0001), caregiver concerns (3.76, p < 0.0001). We also observed that QoL as well as behaviour was independent of age and gender of patients. This study demonstrates significant score changes across all questionnaire domains, comparing pre and post operative data indicating an improvement in their quality of life and behaviour. Hence, we advocate that adenotonsillectomy may be considered as an effective treatment in properly selected patients with OSAS.


Design and Preclinical Validation of the Composite-Type Optical Fiberscope for Minimally Invasive Procedures of Intrauterine Disease.
Because of the rapid growth of minimally invasive surgical procedures, we developed an ultrathin fiberscopic imaging system for laser surgery that could be an important instrument for clinical use in an office setting. A new device, consisting of a custom 1.1-mm diameter flexible fiberscope and ytterbium laser-supported ablation system (composite-type optical fiberscope), has been developed to achieve accurate laser irradiation for minimally invasive procedures of intrauterine disease. This system was validated ex vivo with successful laser ablation under observation. This study provides the design, characterization, performance, and preclinical validation of an optimized composite-type optical fiberscope system. The validation of this fiberscope for endometrial ablation was performed in 12 hysterectomy samples. The irradiated spot diameter and depth were (mean ± SD) 2.80 ± 0.28 mm and 1.53 ± 0.58 mm, respectively, in the range of 185 to 400 J. The effects of the laser on endometrial tissue depended strongly on the combined effects of the output power density and duration of irradiation. The new device makes it possible to perform accurate ablation, because our technology concentrated on combining the 2 subsystems into a single prototype capable of simultaneously observing both the endometrial lesion and the laser spot on a monitor. In conclusion, the feasibility and performance of the composite-type fiberscope system were demonstrated in preclinical studies.


Driver gene detection through Bayesian network integration of mutation and expression profiles.
The identification of mutated driver genes and the corresponding pathways is one of the primary goals in understanding tumorigenesis at the patient level. Integration of multi-dimensional genomic data from existing repositories, e.g., The Cancer Genome Atlas (TCGA), offers an effective way to tackle this issue. In this study, we aimed to leverage the complementary genomic information of individuals and create an integrative framework to identify cancer-related driver genes. Specifically, based on pinpointed differentially expressed genes, variants in somatic mutations and a gene interaction network, we proposed an unsupervised Bayesian network integration (BNI) method to detect driver genes and estimate the disease propagation at the patient and/or cohort levels. This new method first captures inherent structural information to construct a functional gene mutation network and then extracts the driver genes and their controlled downstream modules using the minimum cover subset method. Using other credible sources (e.g. Cancer Gene Census and Network of Cancer Genes), we validated the driver genes predicted by the BNI method in three TCGA pan-cancer cohorts. The proposed method provides an effective approach to address tumor heterogeneity faced by personalized medicine. The pinpointed drivers warrant further wet laboratory validation. The supplementary tables and source code can be obtained from https://xavieruniversityoflouisiana.sharefile.com/d-se6df2c8d0ebe4800a3030311efddafe5. Supplementary data are available at Bioinformatics online.


[The synthesis of cationic glycerolipid acetals containing aliphatic and heterocyclic bases].
1,3-Dioxolane series cationic lipids containing residues of aliphatic or heterocyclic nitrogenous bases were synthesized. The bases were attached to the glycerol backbone either directly (piperazine) or via a spacer group through a thioether bond.


The effect of vaginal delivery and Caesarean section on the anal Sphincter complex of Primipara based on optimized three-dimensional ultrasound image and nuclear regression Reconstruction Algorithm.
The study used the optimized nuclear regression reconstruction algorithm to explore the value of three-dimensional perineal ultrasound evaluation of the effect of caesarean delivery and caesarean section on the anal sphincter complex of primipara. This study performed three-dimensional perineal ultrasound scanning of the anal sphincter complex of 157 primiparas 42 days after delivery. Among them, 77 were in caesarean delivery (spontaneous delivery group) and 80 were in caesarean section (caesarean delivery group) from September 2018 to December 2020 in our hospital. The thickness of the end plane, the middle plane, the distal plane and the distal plane of the external anal sphincter at 3, 6, 9, 12 o'clock direction, and measure the thickness of the central plane of the pubic rectum muscle at 4, 8 o'clock direction. At the same time, the study used tomography and volume contrast imaging to observe the morphology and integrity of the anal sphincter complex. The thickness of the distal anal sphincter at the 12 o'clock direction, the proximal anal sphincter at 6, 12 o'clock, and the central plane at 9 and 12 o'clock in the obstetric group were smaller than those in the caesarean section group (all P < 0.05). There were no significant differences in the thickness of the remaining anal internal and external anal sphincter and puborectalis muscles between the two groups in different directions (all P> 0.05). In the obstetric group, a perineal sphincter defect was found via three-dimensional perineal ultrasound. The delivery method has a certain influence on the shape of the anal sphincter complex. The thickness of the internal and external anal sphincter of the primiparous women in a certain direction is significantly smaller than that of caesarean section. Transperineally three-dimensional ultrasound can clearly show the morphological characteristics and integrity of the anal sphincter complex, and diagnose the defect of the anal sphincter complex.


High alpha-methylacyl-CoA racemase (AMACR) is associated with ERG expression and with adverse clinical outcome in patients with localized prostate cancer.
Alpha-methylacyl-CoA racemase (AMACR) is a well-characterized marker extensively utilized in prostate cancer (PCA) diagnosis. However, the prognostic value of AMACR expression and its relation to TMPRSS2-ERG gene rearrangement as one of the most common molecular alterations in PCA is not fully explored. AMACR expression was investigated in a cohort of 218 men with localized PCA treated by radical prostatectomy and correlated with ERG and various clinical and pathological parameters. In vitro studies assessed AMACR changes to ERG knockdown and other related genes. In addition, bioinformatics validated the significance of AMACR/ERG expression and assessed relevant genetic signatures in relation to AMACR/ERG expression. AMACR expression was significantly associated with disease progression and with ERG (p ∼0). Seventeen percent of cancer foci showed negative/weak AMACR expression while being ERG positive. High AMACR expression was significantly associated with positive surgical margins (p = 0.01), specifically in tumors with lower Gleason score <7, with ∼95 % exhibiting positive surgical margin (p = 0.008). High AMACR showed marginal association with PSA biochemical recurrence (BCR) (p = 0.06) which was slightly more pronounced in ERG-positive tumors (p = 0.04). This was validated in other public cohorts. However, in this cohort, the association with BCR was not statistically significant in multivariate analysis (p = 0.09). Using in vitro cellular models, AMACR messenger RNA (mRNA) expression, but not protein levels, showed an association with ERG expression. We report for the first time a significant association between AMACR and ERG with prognostic implication. Patients with high AMACR/ERG-positive PCA may be at higher risk for disease progression, and additional studies in larger cohorts are needed to confirm the above findings. Functional studies investigating the molecular pathways connecting AMACR and ERG may provide an additional insight into PCA progression pathways.


Accounting for size-specific predation improves our ability to predict the strength of a trophic cascade.
Predation can influence the magnitude of herbivory that grazers exert on primary producers by altering both grazer abundance and their per capita consumption rates via changes in behavior, density-dependent effects, and size. Therefore, models based solely on changes in abundance may miss key components of grazing pressure. We estimated shifts in grazing pressure associated with changes in the abundance and per capita consumption rates of sea urchins triggered by size-selective predation by sea otters (Enhydra lutris). Field surveys suggest that sea otters dramatically decreased the abundance and median size of sea urchins. Furthermore, laboratory experiments revealed that kelp consumption by sea urchins varied nonlinearly as a function of urchin size such that consumption rates increased to the 0.56 and 0.68 power of biomass for red and green urchins, respectively. This reveals that shifts in urchin size structure due to size-selective predation by sea otters alter sea urchin per capita grazing rates. Comparison of two quantitative models estimating total consumptive capacity revealed that a model incorporating shifts in urchin abundance while neglecting urchin size structure overestimated grazing pressure compared to a model that incorporated size. Consequently, incorporating shifts in urchin size better predicted field estimates of kelp abundance compared to equivalent models based on urchin abundance alone. We provide strong evidence that incorporating size-specific parameters increases our ability to describe and predict trophic interactions.


Meta-analysis of the relationship between ACE I/D gene polymorphism and end-stage renal disease in patients with diabetic nephropathy.
Diabetic nephropathy (DN) is the major cause for end-stage renal disease (ESRD) and the pathogenesis for DN developing into ESRD is not clear at present. Results from published studies on the relationship between angiotensin-converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism and ESRD risk in DN patients are still conflicting. This meta-analysis was performed to evaluate the association between ACE I/D gene polymorphism and ESRD risk in DN patients. Association studies were identified from the databases of PubMed, Embase and Cochrane Library on 1 October 2011, and eligible investigations were identified and synthesized using the meta-analysis method. Results were expressed using odds ratios (OR) for dichotomous data and 95% confidence intervals (CI) were also calculated. Twelve studies reporting the relation between ACE I/D gene polymorphism and ESRD risk in DN patients were identified. In overall populations, there was a notable association between D allele or DD genotype and ESRD susceptibility (D: OR = 1.32, 95% CI: 1.11-1.56, P = 0.002; DD: OR = 1.67, 95% CI: 1.25-2.21, P = 0.0004). In the sub-group analysis according to ethnicity, D allele or DD genotype was associated with ESRD risk in Asians. In Caucasians, the association of DD genotype with ESRD risk was observed, but the D allele was not. Furthermore, ACE I/D gene polymorphism was associated with ESRD risk in patients with DN due to diabetes mellitus type 2, but the association was not found for patients with DN due to diabetes mellitus type-1. Our results indicate that D allele or DD homozygous is associated with the ESRD susceptibility in DN patients. However, more investigations are required to further this association.


Real-time vascular imaging and photodynamic therapy efficacy with micelle-nanocarrier delivery of chlorin e6 to the microenvironment of melanoma.
Strategies combining anti-vascular therapy and vascular imaging may facilitate the prediction of early response and outcome in cancer treatment. The aim of this study was to investigate the relationship between the tumor-associated vasculature in melanoma and to develop an approach for melanoma treatment by utilizing the free form and micelle form of the photosensitizer (PS) chlorin e6 in photodynamic therapy (PDT). Green fluorescence protein (GFP) expressing B16-F10 melanoma cells were implanted into the mouse ear dermis. Ce6 in free form or in micelle form was administered via the tail vein. An OV100 imaging system was used to record the red fluorescence of Ce6 to obtain real-time vascular images in the GFP tumor. Compared to free Ce6, Ce6 linked to the micelle-nanocarrier depicted a much clearer vascular image and had an effective vascular destruction by PDT. Micelle Ce6 was localized in lysosomes and in the endoplasmic reticulum of cultured endothelial cells, implying an active endocytosis of the nano-carrier. Micelle Ce6 can serve as a bifunctional PS for vascular imaging and PDT, which facilitates its delivery in the tumor microenvironment.


The spatiotemporal characteristics of the C1 component and its modulation by attention.
Slotnick (this issue) provided a selective review of studies on the attentional modulation of the C1 component of the visual evoked potential, and offers a number of guidelines to maximize the likelihood of observing such modulation in terms of electrode choice, stimulus placement, and types of attentional cue and target stimulus. However, the broader literature pertaining to attentional modulation of the C1 does not support many of these guidelines, and the question of why exactly C1 modulations are so rare remains very much open. Here, we provide clarifications that are critical to an accurate appraisal of the current state of this literature.


Roentogenographic chest screening in the detection and survival of patients with lung cancer. Cooperative Study Group for Early Detection of Lung Cancer in the German Democratic Republic.
In the German Democratic Republic (GDR), annual mass roentgenographic screening of the chest was introduced twenty years ago. To ascertain its value in the detection of lung cancer, data were collected about treatment results at the country's main chest clinics. The study cover 13,283 operations and 10,838 resections, accounting for nearly 90% of all patients with lung cancer treated surgically in the GDR from 1949-1974. From 1965 to 1968, the five-year survival was more favorable in patients who were screened than in those who were diagnosed after clinical symptoms had appeared (36% for the former, 29% for the latter). For improvement of overall results, we recommend differentiated regular chest roentgenographic screening of men 40 to 70 years of age, individualized on the basis of tobacco usage, and full exploitation of all diagnostic and surgical tools now available.


The Inhibitory Innate Immune Sensor NLRP12 Maintains a Threshold against Obesity by Regulating Gut Microbiota Homeostasis.
In addition to high-fat diet (HFD) and inactivity, inflammation and microbiota composition contribute to obesity. Inhibitory immune receptors, such as NLRP12, dampen inflammation and are important for resolving inflammation, but their role in obesity is unknown. We show that obesity in humans correlates with reduced expression of adipose tissue NLRP12. Similarly, Nlrp12-/- mice show increased weight gain, adipose deposition, blood glucose, NF-κB/MAPK activation, and M1-macrophage polarization. Additionally, NLRP12 is required to mitigate HFD-induced inflammasome activation. Co-housing with wild-type animals, antibiotic treatment, or germ-free condition was sufficient to restrain inflammation, obesity, and insulin tolerance in Nlrp12-/- mice, implicating the microbiota. HFD-fed Nlrp12-/- mice display dysbiosis marked by increased obesity-associated Erysipelotrichaceae, but reduced Lachnospiraceae family and the associated enzymes required for short-chain fatty acid (SCFA) synthesis. Lachnospiraceae or SCFA administration attenuates obesity, inflammation, and dysbiosis. These findings reveal that Nlrp12 reduces HFD-induced obesity by maintaining beneficial microbiota.


Synergy and oxygen adaptation for development of next-generation probiotics.
The human gut microbiota has gained interest as an environmental factor that may contribute to health or disease1. The development of next-generation probiotics is a promising strategy to modulate the gut microbiota and improve human health; however, several key candidate next-generation probiotics are strictly anaerobic2 and may require synergy with other bacteria for optimal growth. Faecalibacterium prausnitzii is a highly prevalent and abundant human gut bacterium associated with human health, but it has not yet been developed into probiotic formulations2. Here we describe the co-isolation of F. prausnitzii and Desulfovibrio piger, a sulfate-reducing bacterium, and their cross-feeding for growth and butyrate production. To produce a next-generation probiotic formulation, we adapted F. prausnitzii to tolerate oxygen exposure, and, in proof-of-concept studies, we demonstrate that the symbiotic product is tolerated by mice and humans (ClinicalTrials.gov identifier: NCT03728868 ) and is detected in the human gut in a subset of study participants. Our study describes a technology for the production of next-generation probiotics based on the adaptation of strictly anaerobic bacteria to tolerate oxygen exposures without a reduction in potential beneficial properties. Our technology may be used for the development of other strictly anaerobic strains as next-generation probiotics.


The CDK4/6-UCHL5-BRD4 axis confers resistance to BET inhibitors in MLL-rearranged leukemia cells by suppressing BRD4 protein degradation.
Among acute leukemias, mixed-lineage leukemia-rearranged (MLL-r) leukemia is associated with poor prognosis. Bromodomain and extra-terminal inhibitors (BETi) are promising agents for treatment of hematological malignancies; however, the mechanisms underlying sensitivity to BETi and biomarkers to predict sensitivity are yet to be clarified. Here, we established OTX015-resistant MLL-r cell lines (OTX015-R cells) and used them to explore therapeutic targets in BETi-resistant MLL-r leukemia. OTX015-R cells exhibited resistance to various BETi, and levels of bromodomain-containing protein 4 (BRD4) and BRD4-regulated molecules, such as c-MYC and B-cell/CLL lymphoma-2 (BCL-2), were remarkably increased in OTX015-R cells relative to those in the parental cells; however, BRD4 mRNA transcript levels were not elevated. These results suggest that overexpression of BRD4 protein, through suppression of BRD4 degradation, may contribute to BETi-resistance. Notably, expression of ubiquitin carboxyl-terminal hydrolase isozyme L5 (UCHL5) was increased in OTX015-R cells. Further, a UCHL5 inhibitor, b-AP15, and UCHL5 knockdown had antitumor effects by degrading BRD4. In addition, sensitivity to OTX015 was partially recovered in OTX015-R cells pretreated with b-AP15. Furthermore, cyclin-dependent kinase 4/6 (CDK4/6) inhibition decreased UCHL5 expression, suppressed OTX015-R cell proliferation, and induced apoptosis. These results indicate that the CDK4/6-UCHL5-BRD4 axis confers resistance to BETi by suppressing BRD4 degradation. We propose that this pathway is a potential novel therapeutic target in BETi-resistant MLL-r leukemia with BRD4 overexpression.


Artificial intelligence in bronchopulmonary dysplasia- current research and unexplored frontiers.
Provide an overview of bronchopulmonary dysplasia, its definitions, and their shortcomings. Explore the areas where machine learning may be used to further our understanding of bronchopulmonary dysplasia.


Detection and comparison of phenolic compounds in different extracts of black currant leaves by liquid chromatography coupled with high-resolution ESI-LTQ-Orbitrap MS and high-sensitivity ESI-Qtrap MS.
Liquid preparations such as tinctures and infusion derived from Ribes nigrum L. (black currant) leaves are widely used in Europe. Despite the various uses reported for the infusion and hydroalcoholic extracts of the leaves of this species, their chemical profile has not yet been defined. In the present study, the infusion, hydroalcholic and methanolic extracts prepared from R. nigrum leaves were analyzed. LC-MS analyses of these extracts allowed the characterization of 31 phenolic compounds mainly belonging to organic acids, flavonoids, catechins and its oligomers. Quantitative results proved that these preparations are a rich source of polyphenols and flavonoids and among them the infusion presented the highest polyphenol levels. A metabolomics approach was also used for the comparison and evaluation of the chemical composition of the different extracts obtained from R. nigrum leaves.


Mapping of c-Fos expression in male tree shrew forebrain.
The tree shrew is susceptible to stimuli. However, mapping of c-Fos expression in male tree shrew forebrain has not been explored. The present results provided the first detailed mapping of c-Fos expression in the forebrain of the tree shrew (Tupaia belangeri chinensis). Acute restraint stress rapidly increased the density of c-Fos-immunoreactive (-ir) neurons in the medial orbital cortex (MO), infralimbic cortex, intermediate part of the lateral septal nucleus (LSi), ventral part of the lateral septal nucleus (LSv), anterior part of the bed nucleus of the stria terminalis, posterior part of the bed nucleus of the stria terminalis (STP), paraventricular nucleus of the hypothalamus, supraoptic nucleus, lateral hypothalamic area, ventromedial hypothalamic nucleus (VMH), and medial amygdaloid nucleus (MeA). Furthermore, a significant increase in c-Fos expression was observed in the MO, LSi, LSv, STP, VMH, arcuate hypothalamic nucleus, anterior amygdaloid area, MeA, and cortical amygdaloid nucleus immediately after acute footshock stress. In addition, the distinct patterns of c-Fos expression in the forebrain were shown in context-, restraint-, or footshock-treated tree shrews. In general, the present study provides the first detailed maps of c-Fos expression in male tree shrew forebrain immediately after various stimuli.


Sickness impact in people with muscular dystrophy: a longitudinal study over 10 years.
To describe changes of function in terms of sickness impact over 10 years in adult patients with different types of muscular dystrophy. Patients with muscular dystrophy answered the Sickness Impact Profile and Self-report ADL questionnaires in 1991 and 2001. The study population was identified in a comprehensive prevalence study in the county of Orebro, Sweden. The study group comprised 44 people grouped according to whether they had myotonic dystrophy or muscular dystrophy with proximal or distal muscles affected. Comparison was made between assessments of sickness impact in terms of function at the two time points. Most obvious deterioration over time was in activities of daily living that require finger and arm strength. Ambulation was significantly decreased in myotonic dystrophy and proximal muscular dystrophy. Those walking without assistive devices decreased from 91% to 52%, and the number with a disability pension increased from 36 to 55%. There was a relatively small influence with regard to psychosocial dysfunction assessed by the Sickness Impact Profile. This longitudinal study shows the deteriorating functions reported by patients with muscular dystrophy. This knowledge could be used to formulate new interventions in order to offer appropriate support and treatment to this patient group.


Pros and Cons: Usage of organs from donors infected with hepatitis C virus - Revision in the direct-acting antiviral era.
Should organs from hepatitis C antibody positive donors (HCVD+) be used for transplantation? Organ shortage forces transplant teams to use donors with extended criteria. The decision to transplant a HCVD+ graft is a balance between the risk of transmission of a virus that could lead to end-stage liver diseases and the benefit of access to transplantation, specifically in patients with life-threatening disease. The other issue is the impact of HCV-related liver fibrosis in the donor graft on the long-term outcome in the recipient. Thus, the use of HCVD+ demonstrated a shorter meantime on the waiting list in kidney transplantation. When a HCVD+ graft is transplanted, the risk of HCV transmission depends on; 1) the quality of screening of the donor; 2) the presence of viral replication in the donor at the time of transplantation and the ability to detect it; and 3) the HCV status of the recipient but also the type of transplanted organ. In liver transplantation, the use of HCVD+ graft is usually restricted to recipients with a chronic HCV infection. Several reports showed some competition between HCV donor and recipient strain without deleterious impact on graft and patient survival. Controversies are still pending regarding the quality of the graft and the progression of fibrosis. The recent approval of direct-acting antiviral agents (DAA) dramatically changes the landscape of HCV infection treatment. After transplantation, combinations of DAA show high efficacy and good safety profile. In the near future, extensive use of DAA should reduce the number of HCVD+ with a positive HCV RNA, limiting the risk of transmission but also the number of patients on waiting lists for a disease related to HCV.


Stage 2: Visual information and communication context as modulators of interpersonal coordination in face-to-face and videoconference-based interactions.
Interpersonal coordination of body movement-or similarity in patterning and timing of body movement between interaction partners-is well documented in face-to-face (FTF) conversation. Here, we investigated the degree to which interpersonal coordination is impacted by the amount of visual information available and the type of interaction conversation partners are having. To do so within a naturalistic context, we took advantage of the increased familiarity with videoconferencing (VC) platforms and with limited visual information in FTF conversation due to the COVID-19 pandemic. Pairs of participants communicated in one of three ways: FTF in a laboratory setting while socially distanced and wearing face masks; VC in a laboratory setting with a view of one another's full movements; or VC in a remote setting with a view of one another's face and shoulders. Each pair held three conversations: affiliative, argumentative, and cooperative task-based. We quantified interpersonal coordination as the relationship between the two participants' overall body movement using nonlinear time series analyses. Coordination changed as a function of the contextual constraints, and these constraints interacted with coordination patterns to affect subjective conversation outcomes. Importantly, we found patterns of results that were distinct from previous research; we hypothesize that these differences may be due to changes in the broader social context from COVID-19. Taken together, our results are consistent with a dynamical systems view of social phenomena, with interpersonal coordination emerging from the interaction between components, constraints, and history of the system.


Synthesis of 7- and 12-hydroxy- and 7,12-dihydroxy-3-keto-5 beta-cholan-24-oic acids by reduction of 3,7-, 3,12- and 3,7,12-oxo derivatives.
7 alpha-, 12 alpha-, 12 beta-Hydroxy and 7 alpha,12 alpha- and 7 alpha,12 beta-dihydroxy-3-ketocholanoic acids were prepared in satisfactory yields protecting the 3-keto group as dimethyl ketal and subsequent reduction with sodium borohydride of the corresponding 7- and 12-oxo functionalities. The same procedure gave also 3,12-diketo-7 alpha-hydroxy-cholanoic acid.


Demographic aspects of functional psychoses in Canada.
Canadian national data for functional psychoses (classified as schizophrenia, effective psychoses, paranoid states and reactive psychoses) are analyzed for age, sex, marital status, expectancy for first admissions and length of stay for discharges. Differences are found such that each psychosis can be distinguished from the others, thus providing indirect evidence supporting the use of the different diagnoses. The demographic characteristics of reactive psychoses from North American data have not been previously described, and are found to be similar to Scandinavian descriptions. Sex ratios for subgroup diagnoses whow similarities between catatonic schizophrenics, manic (bipolar) affectives, and reactive psychoses. Schizoaffective psychoses resemble affectives more than schizophrenia, and paraphrenia is similar to affectives. Total expectancies for functional psychosis (4.4% for males, 5.5% for females) are similar to Scandinavian figures, but the distribution by diagnosis differs, perhaps representing different diagnostic practices, but generally similar sex ratios and high rates in single persons are found.


Evidence for an attentional priority map in inferotemporal cortex.
From incoming sensory information, our brains make selections according to current behavioral goals. This process, selective attention, is controlled by parietal and frontal areas. Here, we show that another brain area, posterior inferotemporal cortex (PITd), also exhibits the defining properties of attentional control. We discovered this area with functional magnetic resonance imaging (fMRI) during an attentive motion discrimination task. Single-cell recordings from PITd revealed strong attentional modulation across 3 attention tasks yet no tuning to task-relevant stimulus features, like motion direction or color. Instead, PITd neurons closely tracked the subject's attention state and predicted upcoming errors of attentional selection. Furthermore, artificial electrical PITd stimulation controlled the location of attentional selection without altering feature discrimination. These are the defining properties of a feature-blind priority map encoding the locus of attention. Together, these results suggest area PITd, located strategically to gather information about object properties, as an attentional priority map.


Use of in vitro susceptibility and pathogen prevalence data to model the expected clinical success rates of tigecycline and other commonly used antimicrobials for empirical treatment of complicated skin and skin-structure infections.
The overall expected coverage for tigecycline and selected antimicrobial regimens based upon prevailing susceptibility rates was evaluated for the empirical therapy of complicated skin and skin-structure infections (cSSSIs). Consecutive, non-duplicate bacterial isolates collected from 2000-2005 from patients with documented cSSSI in 57 medical centres located in the USA (38 centres), France (6 centres), Germany (7 centres), Italy (3 centres) and Spain (3 centres) were used to evaluate the frequency of pathogen occurrence and susceptibility rates of select parenteral antimicrobials (SENTRY Program). By applying pathogen-specific susceptibility rates to the frequency of pathogen occurrence in each country, the overall expected coverage for each treatment regimen was measured. The most frequently isolated pathogens were Staphylococcus aureus (33.3-49.9%), Pseudomonas aeruginosa (8.3-17.2%), Escherichia coli (6.6-13.9%) and enterococci (4.1-8.8%). Tigecycline was highly active against the most common pathogens, except for P. aeruginosa and Proteus mirabilis. The highest overall expected empirical coverage was obtained with combination therapy regimens, including vancomycin plus either imipenem (94.4-99.3%) or piperacillin/tazobactam (92.5-98.2%). Among the monotherapy regimens evaluated, tigecycline provided the highest overall expected coverage in the USA (90.6%), France (89.9%) and Italy (83.3%), whilst piperacillin/tazobactam showed the highest expected coverage in Germany (93.1%) and imipenem in Spain (87.7%). Our results suggest that tigecycline represents a viable additional option for the empirical treatment of cSSSI in these countries.


Leucyl-tRNA synthetase regulates lactation and cell proliferation via mTOR signaling in dairy cow mammary epithelial cells.
The role of LeuRS, an aminoacyl-tRNA synthetase, as an intracellular l-leucine sensor for the mTORC1 pathway has been the subject of much research recently. Despite this, the association between LeuRS and lactation in dairy cow mammary epithelial cells (DCMECs) remains unknown. In this study, we found that LeuRS expression in mammary gland tissue was significantly higher during lactation than pregnancy. Moreover, our data demonstrates that LeuRS is localized in the cytoplasm. Treatment with leucine increased DCMECs viability and proliferation, as well as mammalian target of rapamycin (mTOR), p-mTOR, ribosomal protein S6 kinase 1 (S6K1), p-S6K1, β-Casein, sterol regulatory element binding protein 1c (SREBP-1c), glucose transporter 1 (GLUT1), and Cyclin D1 mRNA and protein expression. Secretion of lactose and triglyceride were also increased. siRNA-mediated knockdown of LeuRS led to reduction in all of these processes. Based on these data, LeuRS up-regulates the mTOR pathway to promote proliferation and lactation of DCMECs in response to changes in the intracellular leucine concentration.


Rassf Proteins as Modulators of Mst1 Kinase Activity.
Rassf1A/5 tumor suppressors serve as adaptor proteins possessing a modular architecture with the C-terminal consisting of a coiled-coil SARAH (Salvador-Rassf-Hippo) domain and the central portion being composed of Ras associated (RA) domain. Here, we investigate the effect of Rassf effectors on Mst1 function by mapping the interaction of various domains of Rassf1A/5 and Mst1 kinase using surface plasmon resonance (SPR). The results revealed that apart from the C-terminal SARAH domain of Mst1 which interacts to form heterodimers with Rassf1A/5, the N-terminal kinase domain of Mst1 plays a crucial role in the stabilization of this complex. In addition, SPR experiments show that the RA domains play an important role in fine-tuning the Mst1-Rassf interaction, with Rassf5 being a preferred partner over a similar Rassf1A construct. It was also demonstrated that the activity profile of Mst1 in presence of Rassf adaptors completely switches. A Rassf-Mst1 complexed version of the kinase becomes apoptotic by positively regulating Mst1-H2B mediated serine 14 histone H2B phosphorylation, a hallmark of chromatin condensation. In contrast, the heterodimerization of Mst1 with Rassf1A/5 suppresses the phosphorylation of FoxO, thereby inhibiting the downstream Mst1-FoxO signalling pathway.


Standardization and normative data obtained in the Italian population for a new verbal fluency instrument, the phonemic/semantic alternate fluency test.
The phonemic/semantic alternate fluency test seems to overcome some limits of the instruments currently used to assess set-shifting abilities. In particular, this test does not make high demands on motor systems because the subject is required to rapidly change mental set to generate words by continuously alternating between phonemic and semantic criteria. Thus, it is potentially feasible for use in individuals who have movement disorders. In this regard, some data support its sensitivity in revealing cognitive impairments in people suffering from frontal-striatal-related disorders. The first aim of this study was to provide standardization and normative data for the phonemic/semantic alternate fluency test. The second aim was to upgrade normative data for the single phonemic and semantic fluency tests. For these purposes, we administered to a sample of 335 healthy Italian subjects (ranging from 20 to 90 years), a test consisting of the following three subtests: (1) a single letter-cued (phonemic) fluency subtest; (2) a single category-cued (semantic) subtest; (3) a phonemic/semantic alternate fluency subtest. A composite shifting index was also derived to capture the shifting cost a subject pays passing from performance of the two single fluency subtests to performance of the alternate fluency subtest. We computed correction grids to adjust raw scores for age, literacy and gender according to the results of regression analyses. Moreover, we computed equivalent scores to permit direct and fast comparison of performance on the three fluency tests.


Retrograde autologous priming in cardiopulmonary bypass in adult patients: effects on blood transfusion and hemodilution.
Retrograde autologous priming (RAP) is a cardiopulmonary bypass (CPB) method, at low cost. Previous studies have shown that this method reduces hemodilution and blood transfusions needs through increased intra-operative hematocrit. To evaluate RAP method, in relation to standard CPB (crystalloid priming), in adult patients. Sixty-two patients were randomly allocated to two groups: 1) Group RAP (n = 27) of patients operated using the RAP and; 2) Control group of patients operated using CPB standard crystalloid method (n = 35). The RAP was performed by draining crystalloid prime from the arterial and venous lines, before CPB, into a collect recycling bag. The main parameters analyzed were: 1) CPB hemodynamic data; 2) Hematocrit and hemoglobin values; 3) The need for blood transfusions. It was observed statistically significant fewer transfusions during surgery and reduced CPB hemodilution using RAP. The CPB hemodynamic values were similar, observing a tendency to use lower CPB flows in the RAP group patients. This investigation was designed to be a small-scale pilot study to evaluate the effects of RAP, which were demonstrated concerning the CPB hemodilution and blood transfusions.


Ginsenoside Rh2 attenuates microglial activation against toxoplasmic encephalitis via TLR4/NF-κB signaling pathway.
Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an anti-T. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.


Neuroprotective effects of pyridoxal phosphate and pyridoxal against ischemia in monkeys.
Previously, in monkeys undergoing 20 min whole brain ischemia we demonstrated that the activated calpain-induced lysosomal disruption with the resultant leakage of cathepsins B and L, causes neuronal death in the cornu Ammonis (CA) 1 sector on day 5. Selective cathepsin inhibitors significantly protected ischemic CA1 neurons from delayed necrosis. Recently, pyridoxal phosphate (PLP) and pyridoxal (hydrochloride) (PL) were demonstrated to inhibit cathepsins B and L in vitro, because the active aldehyde at position 4 of the pyridine ring has an affinity for the active site -SH of cysteine residues of cathepsins. Here, we studied whether PLP and PL can, in vivo, protect monkey CA1 neurons from ischemic insult. In monkeys undergoing 20 min whole brain ischemia, 15 mg/kg body weight/day of drugs were intravenously injected for 10 days before and after the ischemic insult. Histological analysis of the surviving CA1 neurons was done using the hippocampus resected on day 5 after ischemia. For PLP or PL, approximately 17% (P = 0.0639) or 54% (P < 0.0001) of the total population (100%) of control CA1 neurons were, respectively, saved from the ischemia-induced neuronal death, showing a remarkable contrast to the surviving neurons (approximately 3.9%) in non-treated monkeys. These data suggested that PL (perhaps PLP intracellularly) is useful as a novel neuroprotectant in primates.


Advances in Genetics and Epigenetic Alterations in Alzheimer's Disease: A Notion for Therapeutic Treatment.
Alzheimer's disease (AD) is a disabling neurodegenerative disorder that leads to long-term functional and cognitive impairment and greatly reduces life expectancy. Early genetic studies focused on tracking variations in genome-wide DNA sequences discovered several polymorphisms and novel susceptibility genes associated with AD. However, despite the numerous risk factors already identified, there is still no fully satisfactory explanation for the mechanisms underlying the onset of the disease. Also, as with other complex human diseases, the causes of low heritability are unclear. Epigenetic mechanisms, in which changes in gene expression do not depend on changes in genotype, have attracted considerable attention in recent years and are key to understanding the processes that influence age-related changes and various neurological diseases. With the recent use of massive sequencing techniques, methods for studying epigenome variations in AD have also evolved tremendously, allowing the discovery of differentially expressed disease traits under different conditions and experimental settings. This is important for understanding disease development and for unlocking new potential AD therapies. In this work, we outline the genomic and epigenomic components involved in the initiation and development of AD and identify potentially effective therapeutic targets for disease control.


Effect of intramolecular fatty acid distribution on aspects of triacylglycerol digestion and absorption studied in vitro.
The triacylglycerols from several natural sources have been used as substrates for in vitro assays of three different aspects of fat digestion and absorption. The triacylglycerol mixtures were chosen because they differed widely both in total fatty acid composition and in the intramolecular distribution of fatty acids. The assays were designed to test the rate of hydrolysis with pancreatic lipase, the ability to form insoluble calcium soaps during the hydrolysis, and the ability to form mixed micelles with bile during the hydrolysis. Differences in the rate of hydrolysis appeared to be related to the structure and the triacylglycerols from lard and human milk, both of which have palmitic acid esterified in the sn-2 position, were hydrolysed most rapidly. Similarly the ability to form calcium soaps was related to structure and those triacylglycerols which released saturated fatty acids on hydrolysis favoured calcium soap formation. While there was a range of abilities to form mixed micelles with bile no obvious correlation with structure was detected.