Role of PLGA Variability in Controlled Drug Release from Dexamethasone Intravitreal Implants.

Long-acting injectable formulations based on poly(lactide-co-glycolide) (PLGA) have been commercialized for over 30 years in at least 20 FDA-approved products. These formulations offer several advantages, including reduced dosing frequency, improved patient compliance, and maintenance of therapeutic levels of drug. Despite extensive studies, the inherent complexity of the PLGA copolymer still poses significant challenges associated with the development of generic formulations having drug release profiles equivalent to those of the reference listed drugs. In addition, small changes to PLGA physicochemical properties or the drug product manufacturing process can have a major impact on the drug release profile of these long-acting formulations. This work seeks to better understand how variability in the physicochemical properties of similar PLGAs affects drug release from PLGA solid implants using Ozurdex (dexamethasone intravitreal implant) as the model system. Four 50:50, acid-terminated PLGAs of similar molecular weights were used to prepare four dexamethasone intravitreal implants structurally equivalent to Ozurdex. The PLGAs were extensively characterized by using a variety of analytical techniques prior to implant manufacture using a continuous, hot-melt extrusion process. In vitro release testing of the four structurally equivalent implants was performed in both normal saline and phosphate-buffered saline (PBS), yielding drastically different results between the two methods. In normal saline, no differences in the release profiles were observed. In PBS, the drug release profiles were sensitive to small changes in the residual monomer content, carboxylic acid end group content, and blockiness of the polymers. This finding further underscores the need for a physiologically relevant in vitro release testing method as part of a robust quality control strategy for PLGA-based solid implant formulations.

Misregulation of cell adhesion molecules in the Ciona neural tube closure mutant bugeye.

Neural tube closure (NTC) is a complex multi-step morphogenetic process that transforms the flat neural plate found on the surface of the post-gastrulation embryo into the hollow and subsurface central nervous system (CNS). Errors in this process underlie some of the most prevalent human birth defects, and occur in about 1 out of every 1000 births. Previously, we discovered a mutant in the basal chordate Ciona savignyi (named bugeye) that revealed a novel role for a T-Type Calcium Channel (Cav3) in this process. Moreover, the requirement for CAV3s in Xenopus NTC suggests a conserved function among the chordates. Loss of CAV3 leads to defects restricted to anterior NTC, with the brain apparently fully developed, but protruding from the head. Here we report first on a new Cav3 mutant in the related species C. robusta. RNAseq analysis of both C. robusta and C. savignyi bugeye mutants reveals misregulation of a number of transcripts including ones that are involved in cell-cell recognition and adhesion. Two in particular, Selectin and Fibronectin leucine-rich repeat transmembrane, which are aberrantly upregulated in the mutant, are expressed in the closing neural tube, and when disrupted by CRISPR gene editing lead to the open brain phenotype displayed in bugeye mutants. We speculate that these molecules play a transient role in tissue separation and adhesion during NTC and failure to downregulate them leads to an open neural tube.

Disruption of left-right axis specification in Ciona induces molecular, cellular, and functional defects in asymmetric brain structures.

BACKGROUND: Left-right asymmetries are a common feature of metazoans and can be found in a number of organs including the nervous system. These asymmetries are particularly pronounced in the simple central nervous system (CNS) of the swimming tadpole larva of the tunicate Ciona, which displays a chordate ground plan. While common pathway elements for specifying the left/right axis are found among chordates, particularly a requirement for Nodal signaling, Ciona differs temporally from its vertebrate cousins by specifying its axis at the neurula stage, rather than at gastrula. Additionally, Ciona and other ascidians require an intact chorionic membrane for proper left-right specification. Whether such differences underlie distinct specification mechanisms between tunicates and vertebrates will require broad understanding of their influence on CNS formation. Here, we explore the consequences of disrupting left-right axis specification on Ciona larval CNS cellular anatomy, gene expression, synaptic connectivity, and behavior. RESULTS: We show that left-right asymmetry disruptions caused by removal of the chorion (dechorionation) are highly variable and present throughout the Ciona larval nervous system. While previous studies have documented disruptions to the conspicuously asymmetric sensory systems in the anterior brain vesicle, we document asymmetries in seemingly symmetric structures such as the posterior brain vesicle and motor ganglion. Moreover, defects caused by dechorionation include misplaced or absent neuron classes, loss of asymmetric gene expression, aberrant synaptic projections, and abnormal behaviors. In the motor ganglion, a brain structure that has been equated with the vertebrate hindbrain, we find that despite the apparent left-right symmetric distribution of interneurons and motor neurons, AMPA receptors are expressed exclusively on the left side, which equates with asymmetric swimming behaviors. We also find that within a population of dechorionated larvae, there is a small percentage with apparently normal left-right specification and approximately equal population with inverted (mirror-image) asymmetry. We present a method based on a behavioral assay for isolating these larvae. When these two classes of larvae (normal and inverted) are assessed in a light dimming assay, they display mirror-image behaviors, with normal larvae responding with counterclockwise swims, while inverted larvae respond with clockwise swims. CONCLUSIONS: Our findings highlight the importance of left-right specification pathways not only for proper CNS anatomy, but also for correct synaptic connectivity and behavior.

Consequences of school closure on access to education: Lessons from the 2013-2016 Ebola pandemic.

The COVID-19 pandemic has seen an unprecedented shutdown of society. Among the various safety measures taken, much attention has been given to school closure as a non-pharmaceutical mitigation tool to curb the spread of the disease through ensuring "social" (physical) distancing. Nearly 1.725 billion children in over 95% of countries worldwide have been affected by school closures implemented in April 2020 as the virus continued to spread. In the field of education, policymakers' attention has been directed at keeping students on board through remote learning and addressing the immediate needs of schools upon reopening. The study presented in this article focuses on who remains absent after schools resume. Using publicly available survey data from the USAID Demographic Health Surveys Program and the UNICEF Multiple Indicator Cluster Survey from before and after the 2013-2016 Ebola pandemic in Guinea and Sierra Leone in West Africa, the author examined changes in school enrolment and dropout patterns, with targeted consideration given to traditionally marginalised groups. At the time, schools closed for between seven to nine months in the two countries; this length and intensity makes this Ebola pandemic the only health crisis in the recent past to come close to the pandemic-related school closures experienced in 2020. The author's findings suggest that post-Ebola, youth in the poorest households saw the largest increase in school dropout. Exceeding expected pre-Ebola dropout rates, an additional 17,400 of the poorest secondary-age youth were out of school. This evidence is important for minimising the likely post-COVID-19 expansion in inequality. The author's findings point to the need for sustainable planning that looks beyond the reopening of educational institutions to include comprehensive financial support packages for groups most likely to be affected.

Conséquences de la fermeture des écoles sur l'accès à l'éducation : les enseignements de la pandémie d'Ebola de 2013 à 2016 ­ La pandémie de COVID-19 a infligé un arrêt sans précédent à la société. Parmi les diverses mesures de sécurité qui ont été prises, une attention particulière à été accordée à la fermeture des écoles devenue un outil non pharmaceutique pour freiner la propagation de la maladie par le biais d'une distanciation « sociale ¼ (physique) ainsi garantie. Près de 1,725 milliard d'enfants dans plus de 95 % des pays du monde entier ont été concernés par les fermetures d'établissements scolaires entrées en vigueur en avril 2020, alors que le virus continuait de se propager. Dans le domaine de l'éducation, les politiques se sont appliquées à permettre aux écoliers de continuer à étudier au moyen de l'apprentissage à distance et à répondre aux besoins immédiats des écoles lors de leur réouverture. L'étude présentée dans cet article est essentiellement consacrée à ceux qui restent absents une fois que l'école a repris. S'appuyant sur des données accessibles au public du Programme d'enquêtes démographiques et sanitaires de l'USAID (l'Agence des États-Unis pour le développement international) et l'Enquête par grappes à indicateurs multiples de l'UNICEF avant et après la pandémie d'Ebola de 2013­2016 en Guinée et en Sierra Leone, en Afrique de l'Ouest, l'auteur s'est penché sur les changements concernant les inscriptions dans les établissements scolaires et les schémas d'abandon de la scolarité en portant une attention particulière aux groupes traditionnellement marginalisés. À cette époque-là, les écoles ont fermé leurs portes pendant sept à neuf mois dans les deux pays. La durée et l'intensité de ces fermetures ont fait de la pandémie d'Ebola la seule crise sanitaire du passé récent à se rapprocher en la matière de celle de COVID-19 en 2020. Les conclusions de l'auteur indiquent qu'après la pandémie d'Ebola, la plus forte hausse des abandons scolaires a été enregistrée chez jeunes des foyers les plus démunis. Dépassant les taux d'abandon d'avant Ebola auxquels on s'attendait, 17 000 écoliers supplémentaires faisant partie des plus pauvres au niveau de l'enseignement secondaire ont abandonné l'école, une preuve importante pour œuvrer à réduire au minimum l'augmentation probable des inégalités après la pandémie de COVID-19. Les conclusions de l'auteur montrent qu'il est nécessaire de planifier durablement en se projetant au-delà de la réouverture des écoles afin d'inclure de vastes aides financières pour les groupes qui seront probablement touchés.

A microRNA-mRNA expression network during oral siphon regeneration in Ciona.

Here we present a parallel study of mRNA and microRNA expression during oral siphon (OS) regeneration in Ciona robusta, and the derived network of their interactions. In the process of identifying 248 mRNAs and 15 microRNAs as differentially expressed, we also identified 57 novel microRNAs, several of which are among the most highly differentially expressed. Analysis of functional categories identified enriched transcripts related to stress responses and apoptosis at the wound healing stage, signaling pathways including Wnt and TGFß during early regrowth, and negative regulation of extracellular proteases in late stage regeneration. Consistent with the expression results, we found that inhibition of TGFß signaling blocked OS regeneration. A correlation network was subsequently inferred for all predicted microRNA-mRNA target pairs expressed during regeneration. Network-based clustering associated transcripts into 22 non-overlapping groups, the functional analysis of which showed enrichment of stress response, signaling pathway and extracellular protease categories that could be related to specific microRNAs. Predicted targets of the miR-9 cluster suggest a role in regulating differentiation and the proliferative state of neural progenitors through regulation of the cytoskeleton and cell cycle.

Thermal Field-Flow Fractionation for Characterization of Architecture in Hyperbranched Aromatic-Aliphatic Polyesters with Controlled Branching.

Thermal field-flow fractionation (ThFFF) was used to characterize the architecture of aromatic-aliphatic polyesters with varying degrees of branching. Thermal diffusion and Soret coefficients (DT and ST, respectively) provide a novel route to polymer architecture analysis. This paper demonstrates an innovative strategy to extract architecture information from the physicochemical separation parameters embedded in ThFFF retention times without explicit separation of linear and branched samples. A Soret contraction factor (g″), defined as the ratio of the ST of a branched polymer to the ST of a molecular weight equivalent linear analogue, is introduced as a metric to indicate degree of branching (DB). This approach circumvents several challenges associated with the analysis of high molar mass polymers with a high degree of branching. The g″ value is shown to be proportional to the degree of branching for linear (DB, 0%), gradually branched (DB, <50%), hyperbranched (DB, 50%), and pseudodendritic (DB, 100%) polyesters allowing the establishment of architecture calibration curves. Furthermore, positive log(g″) values (∼0.2) at low molar mass are attributed to cyclic subpopulations. This work demonstrates the usefulness of the Soret contraction factor for statistically and hyperbranched polymer systems and its sensitivity to cyclic polymers.

Conserved Activity of Reassociated Homotetrameric Protein Subunits Released from Mesoporous Silica Nanoparticles.

Mesoporous silica nanoparticles (MSN) with enlarged pores were prepared and characterized, and reversibly dissociated subunits of concanavalin A were entrapped in the mesopores, as shown by multiple biochemical and material characterizations. When loaded in the MSN, we demonstrated protein stability from proteases and, upon release, the subunits reassociated into active proteins shown through mannose binding and o-phthalaldehyde fluorescence. We have demonstrated a versatile and facile method to load homomeric proteins into MSN with potential applications in enhancing the delivery of large therapeutic proteins.

Photoreceptor specialization and the visuomotor repertoire of the primitive chordate Ciona.

The swimming tadpole larva of Ciona has one of the simplest central nervous systems (CNSs) known, with only 177 neurons. Despite its simplicity, the Ciona CNS has a common structure with the CNS of its close chordate relatives, the vertebrates. The recent completion of a larval Ciona CNS connectome creates enormous potential for detailed understanding of chordate CNS function, yet our understanding of Ciona larval behavior is incomplete. We show here that Ciona larvae have a surprisingly rich and dynamic set of visual responses, including a looming-object escape behavior characterized by erratic circular swims, as well as negative phototaxis characterized by sustained directional swims. Making use of mutant lines, we show that these two behaviors are mediated by distinct groups of photoreceptors. The Ciona connectome predicts that these two behavioral responses should act through distinct, but overlapping, visuomotor pathways, and that the escape behavior is likely to be integrated into a broader startle behavior.

Cellular Processes of Notochord Formation.

This review covers recent advances in our understanding of the cell biology and morphogenesis of the ascidian notochord. In its development, the ascidian notochord undergoes a rapid series of cellular and morphogenic events that transform a group of 40 loosely packed cells in the neurula embryo into a tubular column with central lumen in the larva. The ascidian notochord has been a subject of intensive research in recent years, and particular focus in this review will be on events associated with the development and function of polarized cell properties, and the mechanism of lumen formation.

ACAM, a novel member of the neural IgCAM family, mediates anterior neural tube closure in a primitive chordate.

The neural IgCAM family of cell adhesion molecules, which includes NCAM and related molecules, has evolved via gene duplication and alternative splicing to allow for a wide range of isoforms with distinct functions and homophilic binding properties. A search for neural IgCAMs in ascidians (Ciona intestinalis, Ciona savignyi, and Phallusia mammillata) has identified a novel set of truncated family members that, unlike the known members, lack fibronectin III domains and consist of only repeated Ig domains. Within the tunicates this form appears to be unique to the ascidians, and it was designated ACAM, for Ascidian Cell Adhesion Molecule. In C. intestinalis ACAM is expressed in the developing neural plate and neural tube, with strongest expression in the anterior sensory vesicle precursor. Unlike the two other conventional neural IgCAMs in C. intestinalis, which are expressed maternally and throughout the morula and blastula stages, ACAM expression initiates at the gastrula stage. Moreover, C. intestinalis ACAM is a target of the homeodomain transcription factor OTX, which plays an essential role in the development of the anterior central nervous system. Morpholino (MO) knockdown shows that ACAM is required for neural tube closure. In MO-injected embryos neural tube closure was normal caudally, but the anterior neuropore remained open. A similar phenotype was seen with overexpression of a secreted version of ACAM. The presence of ACAM in ascidians highlights the diversity of this gene family in morphogenesis and neurodevelopment.

The Population Education Transition Curve: Education Gradients Across Population Exposure to New Health Risks.

The salutary effect of formal education on health-risk behaviors and mortality is extensively documented: ceteris paribus, greater educational attainment leads to healthier lives and longevity. Even though the epidemiological evidence has strongly indicated formal education as a leading "social vaccine," there is intermittent reporting of counter-education gradients for health-risk behavior and associated outcomes for certain populations during specific periods. How can education have both beneficial and harmful effects on health, and under which contexts do particular effects emerge? It is useful to conceptualize the influence of education as a process sensitive to the nature, timing of entry, and uniqueness of a new pleasurable and desirable lifestyle and/or product (such as smoking) with initially unclear health risks for populations. Developed herein is a hypothesis that the education gradient comprises multiple potent pathways (material, psychological, cognitive) by which health-risk behaviors are influenced, and that there can be circumstances under which pathways act in opposite directions or are differentially suppressed and enhanced. We propose the population education transition (PET) curve as a unifying functional form to predict shifting education gradients across the onset and course of a population's exposure to new health risks and their associated consequences. Then, we estimate PET curves for cases with prior epidemiological evidence of heterogeneous education gradients with health-risk behaviors related to mass-produced cigarettes in China and the United States; saturated fats, sugar, and processed food diets in Latin America; and HIV infection in sub-Saharan Africa. Each offers speculation on interactions between environmental factors during population exposure and education pathways to health-risk behaviors that could be responsible for the temporal dynamics of PET curves. Past epidemiological studies reporting either negative or positive education gradients may not represent contradictory findings as much as come from analyses unintentionally limited to just one part of the PET process. Last, the PET curve formulation offers richer nuances about educational pathways, macro-historical population dynamics, and the fundamental cause of disease paradigm.

A transiently expressed connexin is essential for anterior neural plate development in Ciona intestinalis.

A forward genetic screen in the ascidian Ciona intestinalis identified a mutant line (frimousse) with a profound disruption in neural plate development. In embryos with the frimousse mutation, the anteriormost neural plate cells, which are products of an FGF induction at the blastula and gastrula stages, initially express neural plate-specific genes but fail to maintain the induced state and ultimately default to epidermis. The genetic lesion in the frimousse mutant lies within a connexin gene (cx-11) that is transiently expressed in the developing neural plate in a temporal window corresponding to the period of a-lineage neural induction. Using a genetically encoded calcium indicator we observed multiple calcium transients throughout the developing neural plate in wild-type embryos, but not in mutant embryos. A series of treatments at the gastrula and neurula stages that block the calcium transients, including gap junction inhibition and calcium depletion, were also found to disrupt the development of the anterior neural plate in a similar way to the frimousse mutation. The requirement for cx-11 for anterior neural fate points to a crucial role for intercellular communication via gap junctions, probably through mediation of Ca(2+) transients, in Ciona intestinalis neural induction.

A one-dimensional model of PCP signaling: polarized cell behavior in the notochord of the ascidian Ciona.

Despite its importance in development and physiology the planar cell polarity (PCP) pathway remains one of the most enigmatic signaling mechanisms. The notochord of the ascidian Ciona provides a unique model for investigating the PCP pathway. Interestingly, the notochord appears to be the only embryonic structure in Ciona activating the PCP pathway. Moreover, the Ciona notochord as a single-file array of forty polarized cells is a uniquely tractable system for the study of polarization dynamics and the transmission of the PCP pathway. Here, we test models for propagation of a polarizing signal, interrogating temporal, spatial and signaling requirements. A simple cell-cell relay cascading through the entire length of the notochord is not supported; instead a more complex mechanism is revealed, with interactions influencing polarity between neighboring cells, but not distant ones. Mechanisms coordinating notochord-wide polarity remain elusive, but appear to entrain general (i.e., global) polarity even while local interactions remain important. However, this global polarizer does not appear to act as a localized, spatially-restricted determinant. Coordination of polarity along the long axis of the notochord requires the PCP pathway, a role we demonstrate is temporally distinct from this pathway's earlier role in convergent extension and intercalation. We also reveal polarity in the notochord to be dynamic: a cell's polarity state can be changed and then restored, underscoring the Ciona notochord's amenability for in vivo studies of PCP.

Whole-organ cell shape analysis reveals the developmental basis of ascidian notochord taper.

Here we use in toto imaging together with computational segmentation and analysis methods to quantify the shape of every cell at multiple stages in the development of a simple organ: the notochord of the ascidian Ciona savignyi. We find that cell shape in the intercalated notochord depends strongly on anterior-posterior (AP) position, with cells in the middle of the notochord consistently wider than cells at the anterior or posterior. This morphological feature of having a tapered notochord is present in many chordates. We find that ascidian notochord taper involves three main mechanisms: Planar Cell Polarity (PCP) pathway-independent sibling cell volume asymmetries that precede notochord cell intercalation; the developmental timing of intercalation, which proceeds from the anterior and posterior towards the middle; and the differential rates of notochord cell narrowing after intercalation. A quantitative model shows how the morphology of an entire developing organ can be controlled by this small set of cellular mechanisms.

The effect of exposure to biomass smoke on respiratory symptoms in adult rural and urban Nepalese populations.

BACKGROUND: Half of the world's population is exposed to household air pollution from biomass burning. This study aimed to assess the relationship between respiratory symptoms and biomass smoke exposure in rural and urban Nepal. METHODS: A cross-sectional study of adults (16+ years) in a rural population (n = 846) exposed to biomass smoke and a non-exposed urban population (n = 802) in Nepal. A validated questionnaire was used along with measures of indoor air quality (PM2.5 and CO) and outdoor PM2.5. RESULTS: Both men and women exposed to biomass smoke reported more respiratory symptoms compared to those exposed to clean fuel. Women exposed to biomass were more likely to complain of ever wheeze (32.0 % vs. 23.5%; p = 0.004) and breathlessness (17.8% vs. 12.0%, p = 0.017) compared to males with tobacco smoking being a major risk factor. Chronic cough was similar in both the biomass and non-biomass smoke exposed groups whereas chronic phlegm was reported less frequently by participants exposed to biomass smoke. Higher PM2.5 levels (≥2 SDs of the 24-hour mean) were associated with breathlessness (OR = 2.10, 95% CI 1.47, 2.99) and wheeze (1.76, 1.37, 2.26). CONCLUSIONS: The study suggests that while those exposed to biomass smoke had higher prevalence of respiratory symptoms, urban dwellers (who were exposed to higher ambient air pollution) were more at risk of having productive cough.

Whole Nervous System Expression of Glutamate Receptors Reveals Distinct Receptor Roles in Sensorimotor Circuits.

The goal of connectomics is to reveal the links between neural circuits and behavior. Larvae of the primitive chordate Ciona are well-suited to make contributions in this area. In addition to having a described connectome, Ciona larvae have a range of readily quantified behaviors. Moreover, the small number of neurons in the larval CNS (∼180) holds the promise of a comprehensive characterization of individual neurons. We present single-neuron predictions for glutamate receptor (GlutR) expression based on in situ hybridization. Included are both ionotropic receptors (AMPA, NMDA, and kainate) and metabotropic receptors. The predicted glutamate receptor expression dataset is discussed in the context of known circuits driving behaviors such as phototaxis, mechanosensation, and looming shadow response. The predicted expression of AMPA and NMDA receptors may help resolve issues regarding the co-production of GABA and glutamate by a subset of photoreceptors. The targets of these photoreceptors in the midbrain appear to express NMDA receptors, but not AMPA receptors. This is in agreement with previous results indicating that GABA is the primary neurotransmitter from the photoreceptors evoking a swimming response through a disinhibition mechanism and that glutamate may, therefore, have only a modulatory action in this circuit. Other findings reported here are more unexpected. For example, many of the targets of glutamatergic epidermal sensory neurons (ESNs) do not express any of the ionotropic receptors, yet the ESNs themselves express metabotropic receptors. Thus, we speculate that their production of glutamate may be for communication with neighboring ESNs, rather than to their interneuron targets.

Deep homologies in chordate caudal central nervous systems.

Invertebrate chordates, such as the tunicate Ciona, can offer insight into the evolution of the chordate phylum. Anatomical features that are shared between invertebrate chordates and vertebrates may be taken as evidence of their presence in a common chordate ancestor. The central nervous systems of Ciona larvae and vertebrates share a similar anatomy despite the Ciona CNS having ~180 neurons. However, the depth of conservation between the Ciona CNS and those in vertebrates is not resolved. The Ciona caudal CNS, while appearing spinal cord-like, has hitherto been thought to lack motor neurons, bringing into question its homology with the vertebrate spinal cord. We show here that the Ciona larval caudal CNS does, in fact, have functional motor neurons along its length, pointing to the presence of a spinal cord-like structure at the base of the chordates. We extend our analysis of shared CNS anatomy further to explore the Ciona "motor ganglion", which has been proposed to be a homolog of the vertebrate hindbrain, spinal cord, or both. We find that a cluster of neurons in the dorsal motor ganglion shares anatomical location, developmental pathway, neural circuit architecture, and gene expression with the vertebrate cerebellum. However, functionally, the Ciona cluster appears to have more in common with vertebrate cerebellum-like structures, insofar as it receives and processes direct sensory input. These findings are consistent with earlier speculation that the cerebellum evolved from a cerebellum-like structure, and suggest that the latter structure was present in the dorsal hindbrain of a common chordate ancestor.

doublesex/mab3 related-1 (dmrt1) is essential for development of anterior neural plate derivatives in Ciona.

Ascidian larvae have a hollow, dorsal central nervous system that shares many morphological features with vertebrate nervous systems yet is composed of very few cells. We show here that a null mutation in the gene dmrt1 in the ascidian Ciona savignyi results in profound abnormalities in the development of the sensory vesicle (brain), as well as other anterior ectodermal derivatives, including the palps and oral siphon primordium (OSP). Although the phenotype of the mutant embryos is variable, the majority have a complete loss of the most anterior structures (palps and OSP) and extensive disruption of sensory structures, such as the light-sensitive ocellus, in the sensory vesicle. dmrt1 is expressed early in the blastula embryo in a small group of presumptive ectodermal cells as they become restricted to anterior neural, OSP and palp fates. Despite the early and restricted expression of dmrt1, we were unable, using several independent criteria, to observe a defect in the mutant embryos until the early tailbud stage. We speculate that the variability and late onset in the phenotype may be due to partially overlapping activities of other gene products.

Comparing health status in Belarus between 2001-10: a novel method using surveys with different response categories.

BACKGROUND: Two population surveys were conducted in Belarus: The Living Conditions, Lifestyle and Health (LLH) in 2001 (n = 2000) and The Health in Times of Transition (HITT) in 2010 (n = 1800). Each survey included a question on health status. The LLH questionnaire provided a 4-point Verbal Response Scale, but the HITT questionnaire used a 5-point scale. When translated into Russian, only two response categories of these scales had identical wording. These differences made a direct comparison of self-reported health status between 2001 and 2010 difficult. METHODS: We conducted a Health Category Response Scale (HCRS) survey in 2010 (n = 570) using a 100ths graduated Visual Analogue Scale (VAS) to understand how the response categories of different scales are perceived by Russian speakers. We implemented the HCRS survey's data to calculate the weighted health status (WHS) for each of the original surveys and to compare health status in Belarus between 2001and 2010. RESULTS: The WHS in Belarus showed a small, but statistically significant, improvement of 2.9 points on a 0-100 scale between 2001 and 2010 (56.2 vs. 59.1). Identical response categories were perceived differently on a 4-point and 5-point VAS. The category 'good' ('Хорошее') measured ∼12 points higher, and the category 'bad/poor' ('Плохое') measured ∼16 points lower, on the 4-point compared with the 5-point VAS. CONCLUSION: Our HCRS survey and novel method enabled a direct comparison of questions with different response options. When applied to the LLH and HITT projects, we concluded that health status in Belarus has improved between 2001 and 2010.

Asymmetrical Flow Field Flow Fractionation for Molar Mass Characterization of Polyethylene Oxide in Abuse-Deterrent Formulations.

High molar mass polyethylene oxide (HM-PEO) is commonly used to enhance the mechanical strength of solid oral opioid drug products to deter abuse. Because the properties of PEO depend on molar mass distribution, accurately determining the molar mass distribution is a necessary part of understanding PEO's role in abuse-deterrent formulations (ADF). In this study, an asymmetrical flow field-flow fractionation (AF4) analytical procedure was developed to characterize PEO polymers with nominal molar masses of 1, 4 or 7 MDa as well as those from in-house prepared placebo ADF. The placebo ADF were manufactured using direct compress or hot-melt-extrusion methods, and subjected to physical manipulation, such as heating and grinding before measurement by AF4 were performed. The molar mass distribution characterized by AF4 revealed that PEO was sensitive to thermal stress, exhibiting decreased molar mass with increased heat exposure. The optimized AF4 method was deemed suitable for characterizing HM-PEO, offering adequate dynamic separation range for PEO with molar mass from 100 kDa to approximately 10 MDa.