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The majority of commercially available monoclonal antibody (mAb) formulations are stabilized with one of three non-ionic surfactants: polysorbate 20 (PS20), polysorbate 80 (PS80), or poloxamer 188 (P188). All three surfactants are susceptible to degradation, which can result in functionality loss and subsequent protein aggregation or free fatty acid particle formation. Consequently, quantitative, and qualitative analysis of surfactants is an integral part of formulation development, stability, and batch release testing. Due to the heterogeneous nature of both polysorbates and poloxamer, online isolation of all the compounds from the protein and other excipients that may disturb the subsequent liquid chromatography with charged aerosol detection (LC-CAD) analysis poses a challenge. Herein, we present an analytical method employing LC-CAD, utilizing a combination of anion and cation exchange columns to completely remove proteins online before infusing the isolated surfactant onto a reversed-phase column. The method allows high throughput analysis of polysorbates within 8 minutes and poloxamer 188 within 12 minutes, providing a separation of the surfactant species of polysorbates (unesterified species, lower esters, and higher esters) and poloxamer 188 (early eluters and main species). Accuracy and precision assessed according to the International Council for harmonisation (ICH) guideline were 96 - 109 % and ≤1 % relative standard deviation respectively for all three surfactants in samples containing up to 110 mg/mL mAb. Subsequently, the method was effectively applied to quantify polysorbate 20 and polysorbate 80 in nine commercial drug products with mAb concentration of up to 180 mg/mL., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Although the phenotype of severe hemophilia has been well studied, there are still knowledge gaps in nonsevere hemophilia., Objectives: The objective of this study was to characterize the clinical bleeding phenotype in nonsevere hemophilia and its association with different factor VIII/IX assessments., Methods: This was a cross-sectional, multicenter study to investigate the bleeding phenotype in adults with nonsevere hemophilia by the number of bleeding and joint bleeding in the past 5 years, a joint score, and the International Society on Thrombosis and Haemostasis bleeding assessment tool (ISTH-BAT). Factor levels were analyzed by 1-stage (lowest in history and at study inclusion) and chromogenic assay (at study inclusion). Patients were enrolled between March 2015 and May 2019., Results: Of the 111 patients (86 with mild and 25 with moderate hemophilia), 57 patients (54.8%) reported any bleeding and 24 (23.1%) any joint bleeding in the past 5 years. A joint score ≥1 was found in 44 patients (41.9%), an ISTH-BAT ≥4 in 100 patients (90.1%), and an ISTH-BAT joint item ≥1 in 50 patients (45.0%). Within the ISTH-BAT, muscle and joint bleeds showed the largest difference between mild and moderate hemophilia. The lowest factor VIII/IX level in patients' history was best associated with bleeding outcomes. Factor was inversely associated with joint bleeds (incidence rate ratio 0.88; 95% CI, 0.79-0.98), joint score, and ISTH-BAT (odds ratios from proportional odds ordinal logistic regression 0.92; 95% CI, 0.87-0.97; and 0.89; 95% CI, 0.86-0.93, respectively)., Conclusion: The occurrence of joint bleeding differentiated persons with mild and moderate hemophilia. The ISTH-BAT and lowest factor in patients' history provided valuable information of the bleeding phenotype in nonsevere hemophilia., (© 2023 The Authors.)

The increase in world energy consumption, and the worries from potential future disasters that may derive from climate change have stimulated the development of renewable energy technologies. One promising method is the utilization of whole photosynthetic cyanobacterial cells to produce photocurrent in a bio-photo electrochemical cell (BPEC). The photocurrent can be derived from either the respiratory or photosynthetic pathways, via the redox couple NADP + /NADPH mediating cyclic electron transport between photosystem I inside the cells, and the anode. In the past, most studies have utilized the fresh-water cyanobacterium Synechocystis sp. PCC 6803 (Syn). Here, we show that the globally important marine cyanobacterium Trichodesmium erythraeum flourishing in the subtropical oceans can provide improved currents as compared to Syn. We applied 2D-fluorescence measurements to detect the secretion of NADPH and show that the resulting photocurrent production is enhanced by increasing the electrolyte salinity, Further enhancement of the photocurrent can be obtained by the addition of electron mediators such as NAD + , NADP + , cytochrome C, vitamin B1, or potassium ferricyanide. Finally, we produce photocurrent from additional cyanobacterial species: Synechocystis sp. PCC6803, Synechococcus elongatus PCC7942, Acaryochloris marina MBIC 11017, and Spirulina, using their cultivation media as electrolytes for the BPEC., Competing Interests: Declaration of competing interest The authors declare that they have no competing or financial interests., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Data on the effect of ABO blood group (ABO), von Willebrand factor (VWF) levels, and age on factor VIII (FVIII) in non-severe haemophilia A (HA) is scarce., Objective: To investigate if ABO, VWF levels, and age have an influence on the variability of FVIII levels and consequently on the assessment of severity in non-severe HA., Patients/methods: Eighty-nine patients with non-severe HA and 82 healthy controls were included. Data on ABO was collected and FVIII clotting activity (FVIII:C) with one-stage clotting assay (FVIII:C OSA) and chromogenic substrate assay (FVIII:C CSA), FVIII antigen (FVIII:Ag) and VWF antigen (VWF:Ag) and activity (VWF:Act) were determined., Results: In HA, FVIII:C OSA and CSA and FVIII:Ag were not different between non-O (n = 42, median 15.5, interquartile range 10.4-24.0; 10.0, 6.8-26.0 and 15.2, 10.7-24.9) and O (n = 47, 14.1, 9.0-23.0; 10.0, 5.0-23.0 and 15.2, 9.3-35.5), whereas in healthy controls, non-O individuals had significantly higher FVIII levels., Fviii: C showed no relevant correlation with VWF levels in HA, but we observed strong correlations in healthy controls. Age had only a minor influence in HA, but had a considerable impact on FVIII:C in healthy controls. In multivariable regression analysis ABO, VWF:Ag and age were not associated with FVIII:C in HA, whereas this model explained 61.3% of the FVIII:C variance in healthy controls., Conclusions: We conclude that in non-severe HA ABO and VWF levels do not substantially influence the variability of FVIII levels and age has only minor effects on it, which is important information for diagnostic procedures., (© 2020 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals, Inc. on behalf of International Society on Thrombosis and Haemostasis.)

Porting is a powerful methodology to recalibrate an existing database of ion chromatography (IC) retention times by reflecting the changes of column behavior resulting from either batch-to-batch variability in the production of the column or the manufacture of new versions of a column. This approach has been employed to update extensive databases of retention data of inorganic and organic anions forming part of the "Virtual Column" software marketed by Thermo Fisher Scientific, which is the only available commercial optimization tool for IC separation. The current porting process is accomplished by performing three isocratic separations with two representative analyte ions in order to derive a porting equation which expresses the relationship between old and new data. Although the accuracy of retention prediction is generally enhanced on new columns, errors were observed on some columns. In this work, the porting methodology was modified in order to address this issue, where the porting equation is now derived by using six representative analyte ions (chloride, bromide, iodide, perchlorate, sulfate, and thiosulfate). Additionally, the updated porting methodology has been applied on three Thermo Fisher Scientific columns (AS20, AS19, and AS11HC). The proposed approach showed that the new porting methodology can provide more accurate and robust retention prediction on a wide range of columns, where average errors in retention times for ten test anions under three eluent conditions were less than 1.5%. Moreover, the retention prediction using this new approach provided an acceptable level of accuracy on a used column exhibiting changes in ion-exchange capacity., (Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.)

The relative performance of six multivariate data analysis methods derived from or combined with partial least squares (PLS) has been compared in the context of quantitative structure-retention relationships (QSRR). These methods include, GA (genetic algorithm)-PLS, Monte Carlo uninformative variable elimination (MC-UVE), competitive adaptive reweighted sampling (CARS), iteratively retaining informative variables (IRIV), variable iterative space shrinkage approach (VISSA) and PLS with automated backward selection of predictors (autoPLS). A set of 825 molecular descriptors was computed for 86 suspected sports doping compounds and used for predicting their gradient retention times in reversed-phase liquid chromatography (RPLC). The correlation between molecular descriptors selected by each technique and the retention time was established using the PLS method. All models derived from a selected subset of descriptors outperformed the reference PLS model derived from all descriptors, with very small demands of computational time and effort. A performance comparison indicated great diversity of these methods in selecting the most relevant molecular descriptors, ranging from 28 for CARS to 263 for MC-UVE. While VISSA provided the lowest degree of over-fitting for the training set, CARS demonstrated the best compromise between the prediction accuracy and the number of selected descriptors, with the prediction error of as low as 46s for the external test set. Only ten descriptors were found to be common for all models, with the characteristics of these descriptors being representative of the retention mechanism in RPLC., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In the United States, the generally non-pathogenic trypanosome of cattle is designated Trypanosoma (Megatrypanum) theileri and is distinguished morphologically from Trypanosoma (M.) cervi, a trypanosome originally described in mule deer and elk. Phylogenetic studies of the Megatrypanum trypanosomes using various molecular markers reveal two lineages, designated TthI and TthII, with several genotypes within each. However, to date there is very limited genetic data for T. theileri, and none for the Megatrypanum trypanosomes found in wild ungulates, in the U.S. In this study U.S. isolates from cattle (Bos taurus), white-tailed deer (Odocoileus virginianus) (WTD), and elk (Cervus elaphus canadensis) were compared by ribosomal DNA (rDNA) sequence analysis and their incidence in cattle and WTD in south Texas counties was investigated. Phylogenetic analyses showed clear separation of the bovine and cervine trypanosomes. Both lineages I and II were represented in the U.S. cattle and WTD parasites. Lineage I cattle isolates were of a previously described genotype, whereas WTD and elk isolates were of two new genotypes distinct from the cattle trypanosomes. The cattle isolate of lineage II was of a previously reported genotype and was divergent from the WTD isolate, which was of a new genotype. In La Salle, Starr, Webb, and Zapata counties in south Texas a total of 51.8% of white-tailed deer were positive for trypanosomes by 18S rDNA PCR. Of the cattle screened in Webb County, 35.4% were positive. Drought conditions prevailing in south Texas when the animals were screened suggest the possibility of a vector for Trypanosoma other than the ked (Lipoptena mazamae) and tabanid flies (Tabanus spp. and Haematopota spp.)., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In analogy to our previous publication, the hydrophilic interaction liquid chromatography mechanism was examined in terms of hydrogen bonding, coulombic interactions and phase ratio using linear solvation energy relationships. At first, 23 commercially available and in-house synthesized chromatographic supports are discussed in order to obtain system constants at pH 5.0 with ammonium acetate as buffer salt. Subsequently we compared these outcomes with our former results obtained at pH 3.0 with ammonium formate as buffer additive. Goodness of fit in terms of the adjusted multiple correlation coefficient was found to be reduced under the new conditions. No universal model which simultaneously comprised acidic, basic and neutral analytes could be performed. A significant enhancement of the HILIC systems hydrogen bond basicity was found when changing the pH and buffer counter ions. Even though packing materials showed similar selectivity profiles during the collection of the experimental retention data, different forces were found to account for the overall retention (e.g. Shiseido PC HILIC and Nucleodur HILIC). This indicates that HILIC type selectivity is rather based on a sum of additive or multiplicative phenomena., (Copyright © 2013 Elsevier B.V. All rights reserved.)

22 commercially available and home-made stationary phases with different surface modifications were compared under hydrophilic interaction liquid chromatographic (HILIC) conditions. The column set comprised neutral, basic, acidic, zwitterionic and mixed surface modifications. Retention data of 68 differently structured test solutes were acquired to generate retention models based on a linear solvation energy relationship (LSER) approach. A recently modified solvation parameter model with two additional molecular descriptors was evaluated in terms of its universal applicability when electrostatic forces are enabled in addition to predominant partition phenomena. The suggested method could not be confirmed to be a standardized way to characterize HILIC systems when different operating conditions are applied. However, the significant contribution of the recently introduced charge descriptors (D⁻ and D⁺) on explaining the interactions within HILIC systems was confirmed. The solvation parameter model was found to be a useful tool in the course of column development, to affirm or dismiss the preceding educated guess on how certain immobilized ligands will behave. Acidic modified surfaces (stationary phases) exhibit a very small hydrogen bond acceptor property and are less versatile when it comes to an even distribution of solutes along the retention window. Furthermore, the results indicate that basic and neutral columns are more preferable for HILIC applications and might explain why only a limited variety of strong acidic modified HILIC columns, although found in literature, are available commercially., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Unlabelled: Ultrasonography (USG) is the preferred screening method for fetal brain examination. It has some technical limitations and a relatively low sensibility and specificity for many central nervous system (CNS) malformations. Fetal cerebral magnetic resonance imaging (MRI) offers better resolution and sensibility, with scarce limitations., Objectives: To determine the fetal age according to cortical maturation as seen in MRI, correlating these data with those obtained by means of USG measurements; to correlate USG pathological findings with the MRIs and to determine how the sequence of cortical maturation varies in abnormal brains., Materials and Methods: 50 pregnant women were submitted to USG and fetal brain MRI. Fifteen carried out normal pregnancies. In the remaining 35, the USG, the clinical assessment or both, raised the suspicion of a CNS malformation. Facts studied were: the gestational age calculated by USG, analysis of the cortical gyral development by MRI (cortical age), the presence of CNS abnormalities and the correlation between the cortical maturation and the presence of CNS pathologies. Statistical analysis included the Student's t test for paired samples, the Pearson's correlation coefficient (r) and linear regression curves., Results: In the control group, fetal age highly correlated with the cortical age estimated by MRI. In the abnormal group, a wide variety of pathologies could be found, with higher sensibility and specificity than USG when applying MRI techniques. Cortical age did not correlate with the gestational age in this group; moreover, its estimation could not be achieved in severely malformed brains., Discussion: MRI allows a detailed study of the CNS before birth. It proved to be more reliable and specific than USG, with fewer technical limitations. Cortical maturation can be accurately assessed by this method in normal or slightly abnormal fetuses. However, USG is better than MRI for diagnosing skull bony defects.

Fetal brain Magnetic Resonance Imaging (MRI) is a new technique of growing interest, with a high potential to detect prenatal central nervous system abnormalities. This requires an accurate knowledge of the normal morphological sequence of brain development. In this paper we studied the cortical development of post-mortem normal fetal brains, correlating MRI estimations of fetal age with in vitro anatomical and anthropometric measurements. Ten post-mortem fetal heads were submitted to MRI. Maturational state of sulci and gyri and gray-white matter differentiation were analysed in the MRIs and by dissection of the brains. The findings were correlated with the previously estimated ages of the fetuses, which varied between 17 and 38 weeks. Consistency between methods was assessed employing intraclass correlation coefficient and Bland-Altman plots, with a 95% confidence interval. Estimations of fetal age obtained by MRI were very similar to those achieved by anthropometric measurements or by considering anatomical parameters. Gyral development proved to be more precise than gray-white matter differentiation for this purpose. Fetal MRI proved to be as reliable as the macroscopic anatomical examination for depicting normal cortical developmental sequence and age, suggesting that this technique may be a suitable option for achieving precise information about the morphology of human brains along the gestational period.

Using DNA molecules to construct a structural scaffold for nanotechnology is largely accepted. In this article, we report on two methods for constructing a figure-eight structure of DNA molecules having a relatively high yield that could be used further as a scaffold for nanotechnology applications. In the first method, two plasmids were constructed that, on digestion with a restriction endonuclease producing nicks in the corresponding sites and after heating, produced complementary single-stranded sequences, enabling the plasmids to hybridize to each other and forming a figure-eight structure. The formation of the figure-eight structure was analyzed by restriction analysis and gel electrophoresis as well as by atomic force microscopy. The second method makes use of the bacteriophage M13 that is obtained as either a single- or double-stranded circular DNA molecule. Two M13 molecules harboring complementary sequences were constructed and produced a figure-eight structure on hybridization. The methods described here could be used further for the construction of nanoelectronic devices.

Objectives: We investigated the association between the factor V Leiden gene variant and carotid atherosclerosis in a cross-sectional study and explored possible associations between this gene variant and coronary artery disease (CAD) in a case-control study., Methods: The presence (n=1696) or absence (n=703) of carotid atherosclerosis were sonographically assessed among participants of the population-based Study of Health in Pomerania (SHIP). The case-control study included 1021 patients with severe CAD and 2791 healthy SHIP participants. The factor V Leiden gene variant was determined by PCR and MnlI digestion., Results: Multivariable analyses revealed no independent association between the factor V Leiden gene variant per se and carotid atherosclerosis or CAD. In the cross-sectional study, there was an interaction between the factor V Leiden gene variant and serum LDL cholesterol in non-diabetics with respect to the risk of carotid atherosclerosis. In the case-control study a similar interaction was found for CAD. In both studies the atherosclerotic risk increased with rising serum LDL cholesterol concentrations in carriers of the factor V Leiden gene variant., Conclusion: The co-existence between the factor V Leiden gene variant and high serum LDL cholesterol is independently associated with the risk of atherosclerosis.

Oral cancer is one of the most disfiguring types of cancer, since the surgical removal of the tumor may result in facial distortion. Oral cancer is also known to exhibit "field cancerization", resulting in the development of a second primary tumor. Furthermore, the five-year survival rate of this disease has remained approximately 50% during the past 30 years. Prevention and early detection/treatment of oral cancer could significantly improve the quality of life for individuals at risk. Recently, the targeted elimination of oral squamous cell carcinoma cells by inducing apoptosis has emerged as a valued strategy to combat oral cancer. Studies utilizing a variety of chemical or biological interventions demonstrated promising results for induction of apoptosis in oral malignant cells. This review summarizes the results of a number of investigations focused specifically on induction of apoptosis in oral cancer cells by synthetic compounds and naturally occurring chemopreventive agents with apoptotic potential.

Background: Magnetic Resonance Imaging (MRI) allows a detailed "in vivo" macroscopic study of the human brain; previously, it has been demonstrated that Fluid Attenuated Inversion Recovery (FLAIR) sequence shows higher signal intensity of cortices belonging to limbic structures., Purpose: To measure and compare signal intensities (SI) of cytoarchitectonically different cortical regions., Methods: In 22 adult subjects, without psychiatric or neurological diseases, FLAIR sequence was performed in coronal slices, perpendicular to the main hippocampal axis. Signal intensity was measured, with a region-of-interest (ROI) function, in 12 different cortical regions. We compared these values and grouped the cortices into five groups: (1) limbic cortices, (2) paralimbic agranular cortices, (3) paralimbic granular cortices, (4) parietal-type neopallium, (5) frontal-type neopallium. A t-test for comparison of paired samples was performed, considering p

Class III malocclusion is one of the most difficult anomalies to understand. Because not all Class III patients are candidates for surgical correction, patient assessment and selection remain main issues in diagnosis and treatment planning. The purpose of this study was to separate Class III patients who can be properly treated orthodontically from those who require orthognathic surgery. A large sample size was a necessary to obtain a sufficiently robust model. Thus, a multicentric study design was chosen (Orthodontic Departments of the Universities of Frankfurt, Heidelberg, and Würzburg, Germany). The cephalograms of 175 adult patients with Class III malocclusions were analyzed. The orthodontic group comprised 87 patients, and the surgery group, 88 patients. Twenty linear, proportional, and angular measurements were made. Stepwise discriminant analysis was applied to identify the dentoskeletal variables that best separate the groups. The discriminant function model was highly significant (P <.0001); 92% of the patients were correctly classified. The following variables were extracted: Wits appraisal, length of the anterior cranial base, maxillary/mandibular (M/M) ratio, and lower gonial angle. The resulting equation was: Individual score = -1.805 + 0.209. Wits + 0.044. S-N + 5.689. M/M ratio - 0.056. Go(lower). By means of discriminant analysis, correct classification of adult Class III malocclusion patients succeeded to a very high degree. Of all the variables, the Wits appraisal was the most decisive parameter.

Dihydroorotic acid hydrazide (DHOH) did not inhibit the replication of potatovirus X (PVX) in leaf disks of Nicotiana tabacum 'Samsun'. In contrast, 5-fluoroorotic acid (5-FOA) completely inhibited the replication of PVX, as demonstrated by a serological virus assay as well as a local lesion bioassay using Gomphrena globosa as the test plant. The corresponding base analogue 5-fluorouracil (5-FU) had only a weak inhibitory effect. Time-course inhibition experiments in synchronized virus-infected leaf disks led us to conclude that 5-FOA, as well as 5-azadihydrouracil (5-ADHU), an uracil catabolite analogue, inhibit the same early event in virus infection. Neither 5-FOA nor 5-ADHU had a direct inactivating effect on free PVX virions.

9-(2,3-Dihydroxypropyl)adenine (DHPA) and 3-(adenin-9-yl)-2-hydroxypropanoic acid 2-methylpropylester (AHPA-MP)) markedly inhibit the replication of potato virus X (PVX). The latter compound inhibits virus replication more effectively at the lower concentration range. In synchronized virus-infected leaf-disks, the time-response course of inhibition by both compounds is almost identical, and similar to that of cycloheximide.

Inhibition of potato virus X replication can be enhanced by complexation of N-allyl-N'-2-pyridyl-thiourea and of N-phenyl-N'-2-pyridyl-thiourea with Cu+, Zn2+ and Cd2+. The antiphytoviral activity of some complexes with pyridylthiourea can be further augmented by adding small quantities of 2,4-dioxohexahydro-1,3,5-triazine.