Nowadays, most of the newly developed active pharmaceutical ingredients (APIs) consist of cohesive particles with a mean particle size of <100μm, a wide particle size distribution (PSD) and a tendency to agglomerate, therefore they are difficult to handle in continuous manufacturing (CM) lines. The current paper focuses on the impact of various glidants on the bulk properties of difficult-to-handle APIs. Three challenging powders were included: two extremely cohesive APIs (acetaminophen micronized (APAPμ) and metoprolol tartrate (MPT)) which previously have shown processing issues during different stages of the continuous direct compression (CDC)-line and a spray dried placebo (SD) powder containing hydroxypropylmethyl cellulose (HPMC), known for its sub-optimal flow with a high specific surface area (SSA) and low density. Four flow-enhancing excipients were used: a hydrophilic (Aerosil® 200) and hydrophobic (Aerosil® R972) fumed silica grade, a mesoporous silica grade (Syloid® 244FP), and a calcium phosphate excipient (TRI-CAFOS® 200-7). The APIs and binary API/glidant blends (varied between 0.5-2.75 w/w%) were characterized for their bulk properties relevant for CDC. The results indicated that optimizing different bulk parameters (e.g., density, flow, compressibility..) of an API required varying weight percentages of the glidant (e.g., different surface area coverage (SAC)) depending on the APIs. Moreover, even at similar SAC, the impact of the glidant on the bulk characteristic of the APIs depended on the glidant type properties. While nano-sized silicon dioxide were effective for improving the flowability of a powder, other glidants (mesoporous silica and tricalcium phosphate (TCP)) showed also promise as alternatives. Additionally, an excess of glidant, referred to as oversilication, negatively impacted some bulk parameters, but other characteristics were unaffected. Finally, to determine the appropriate concentration of the different classes of glidants, SAC calculations, an understanding of the glidant's working mechanism, and knowledge about the API's characteristics (i.e., morphology, compressibility, flowability, aeration, density, and wall friction) are required. This study confirmed the necessity of including various material characterization techniques to assess the impact of glidants on the bulk characteristics of APIs.
- MeSH
- Hypromellose Derivatives * chemistry MeSH
- Chemistry, Pharmaceutical methods MeSH
- Calcium Phosphates * chemistry MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Metoprolol * chemistry MeSH
- Bulk Drugs MeSH
- Silicon Dioxide chemistry MeSH
- Acetaminophen * chemistry MeSH
- Excipients * chemistry MeSH
- Powders * MeSH
- Drug Compounding methods MeSH
- Rheology * MeSH
- Particle Size * MeSH
- Publication type
- Journal Article MeSH
The interaction between the main psychotropic ingredient of Cannabis, Δ9- tetrahydrocannabinol (THC), with the endogenous cannabinoid system (ECS) is a critical and underrated issue that deserves utmost attention. The ECS, indeed, contributes to the formation and regulation of excitatory and inhibitory (E/I) neuronal networks that in the hippocampus underly spatial memory. This study explored sex-specific consequences of prenatal exposure to THC in hippocampus-dependent memory and the underlying cellular and molecular contributors of synaptic plasticity and E/I homeostasis. Sprague Dawley dams were exposed to THC (2 mg/kg) or vehicle, from gestational day 5-20. The adolescent progeny of both sexes was tested for: spatial memory retrieval and flexibility in the Barnes Maze; mRNA expression of relevant players of hippocampal synaptic plasticity; density of cholecystokinin-positive basket cells (CCK+BCs) - a major subtype of hippocampal inhibitory interneurons; mRNA expression of the excitatory and inhibitory synaptic proteins neuroligins (Nlgns), as a proxy of synaptic efficiency. Our results show a sex-specific disruption in spatial memory retrieval and flexibility, a male-specific decrease in CCK+BCs density and increase in the expression of markers of neuroplasticity, and consistent changes in the expression of Nlgn-1 and 3 isoforms. Despite a delay in memory retrieval, flexibility of memory was spared in prenatally-THC-exposed female offspring as well as most of the markers of neuroplasticity; a sex-specific increase in CCK+BCs density, and a consistent expression of Nlgn-3 was observed. The current results highlight a major vulnerability to prenatal exposure to THC on memory processing in the male progeny, and sex-specific alterations in the E/I balance and synaptic plasticity.
- MeSH
- Maze Learning drug effects MeSH
- Cholecystokinin metabolism MeSH
- Hippocampus * drug effects metabolism MeSH
- Rats MeSH
- Neuronal Plasticity * drug effects MeSH
- Sex Characteristics * MeSH
- Rats, Sprague-Dawley * MeSH
- Spatial Memory * drug effects MeSH
- Pregnancy MeSH
- Dronabinol * pharmacology toxicity MeSH
- Prenatal Exposure Delayed Effects * chemically induced MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Pregnancy MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
It has long been known that environmental conditions, particularly during development, affect morphological and functional properties of the brain including sensory systems; manipulating the environment thus represents a viable way to explore experience-dependent plasticity of the brain as well as of sensory systems. In this review, we summarize our experience with the effects of acoustically enriched environment (AEE) consisting of spectrally and temporally modulated complex sounds applied during first weeks of the postnatal development in rats and compare it with the related knowledge from the literature. Compared to controls, rats exposed to AEE showed in neurons of several parts of the auditory system differences in the dendritic length and in number of spines and spine density. The AEE exposure permanently influenced neuronal representation of the sound frequency and intensity resulting in lower excitatory thresholds, increased frequency selectivity and steeper rate-intensity functions. These changes were present both in the neurons of the inferior colliculus and the auditory cortex (AC). In addition, the AEE changed the responsiveness of AC neurons to frequency modulated, and also to a lesser extent, amplitude-modulated stimuli. Rearing rat pups in AEE leads to an increased reliability of acoustical responses of AC neurons, affecting both the rate and the temporal codes. At the level of individual spikes, the discharge patterns of individual neurons show a higher degree of similarity across stimulus repetitions. Behaviorally, rearing pups in AEE resulted in an improvement in the frequency resolution and gap detection ability under conditions with a worsened stimulus clarity. Altogether, the results of experiments show that the exposure to AEE during the critical developmental period influences the frequency and temporal processing in the auditory system, and these changes persist until adulthood. The results may serve for interpretation of the effects of the application of enriched acoustical environment in human neonatal medicine, especially in the case of care for preterm born children.
- MeSH
- Acoustic Stimulation * MeSH
- Acoustics MeSH
- Inferior Colliculi growth & development physiology MeSH
- Rats MeSH
- Humans MeSH
- Neurons physiology MeSH
- Neuronal Plasticity * physiology MeSH
- Animals, Newborn MeSH
- Auditory Pathways * growth & development physiology MeSH
- Auditory Perception MeSH
- Auditory Cortex * growth & development physiology MeSH
- Age Factors MeSH
- Environment MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Osteomalacie patří mezi metabolické osteopatie a je charakterizována poruchou mineralizace nově tvořeného osteoidu. Etiologie onemocnění je široká a zahrnuje vrozené a získané příčiny. Tyto příčiny vedou většinou k chybění substrátů mineralizace (vápník, fosfáty) nebo k porušenému procesu mineralizace. V praxi může být klinický obraz osteomalacie neúplný a laboratorní obraz může zahrnovat širokou škálu odchylek. Z těchto důvodů může být diagnóza osteomalacie stanovena opožděně nebo zaměněna za jinou patologii skeletu. Osteomalacie patří rutinně do diferenciální diagnostiky při nálezu nízké kostní denzity, i když se s ní ve srovnání s osteoporózou setkáváme v klinické praxi vzácněji. Léčba onemocnění závisí na vyvolávající příčině a při správné diagnostice může dojít k plnému vyléčení. Prezentujeme kazuistiku pacienta ze standardního lůžkového interního oddělení, u kterého nebyla diagnóza osteomalacie jednoznačná. Kazuistika popisuje 75letého kachektického pacienta s malnutriční osteomalacií při nepoznané chronické pankreatitidě.
Osteomalacia belongs to the metabolic osteopathies and is characterized by a mineralization disorder of the newly formed osteoid. The etiology of the disease is broad and includes congenital and acquired causes. These causes usually lead to a lack of mineralization substrates (calcium, phosphates) or to an impaired mineralization process. In practice, the clinical picture of osteomalacia can be incomplete, and the laboratory picture can include a wide range of deviations. For these reasons, the diagnosis of osteomalacia may be delayed or mistaken for another skeletal pathology. Osteomalacia is routinely included in the differential diagnosis when low bone density is found, although it is less common in clinical practice compared to osteoporosis. The treatment of the disease depends on the underlying cause, and with the correct diagnosis, a full cure can occur. We present a case report of a patient from a standard inpatient ward, in whom the diagnosis of osteomalacia was not clear. The case report includes a 75-year-old cachectic patient with malnourished osteomalacia with unrecognized chronic pancreatitis.
- MeSH
- Pancreatitis, Chronic diagnosis complications MeSH
- Cachexia etiology complications MeSH
- Humans MeSH
- Malabsorption Syndromes etiology complications MeSH
- Osteomalacia * etiology physiopathology MeSH
- Aged MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Aged MeSH
- Publication type
- Case Reports MeSH
The advent of new, advanced quantitative MRI metrics allows for in vivo evaluation of multiple biological processes highly relevant for ageing. The presented study combines several MRI parameters hypothesised to detect distinct biological characteristics as myelin density, cellularity, cellular membrane integrity and iron concentration. 116 healthy volunteers, continuously distributed over the whole adult age span, underwent a multi-modal MRI protocol acquisition. Scatterplots of individual MRI metrics revealed that certain MRI protocols offer much higher sensitivity to early adulthood changes while plateauing in higher age (e.g., global functional connectivity in cerebral cortex or orientation dispersion index in white matter), while other MRI metrics provided reverse ability-stable levels in young adulthood with sharp changes with rising age (e.g., T1ρ and T2ρ). Nonetheless, despite the previously published validations of specificity towards microstructural biology based on cytoarchitectonic maps in healthy population or alterations in certain pathologies, several metrics previously hypothesised to be selective to common measures failed to show similar scatterplot distributions, pointing to further confounding factors directly related to age. Furthermore, other metrics, previously shown to detect different biological characteristics, exhibited substantial intercorrelations, be it due to the nature of the MRI protocol itself or co-dependence of relevant biological microstructural processes. All in all, the presented study provides a unique basis for the design and choice of relevant MRI parameters depending on the age group of interest. Furthermore, it calls for caution in simplistic biological inferences in ageing based on one simple MRI metric, even though previously validated under other conditions. Complex multi-modal approaches combining several metrics to extract the shared subcomponent will be necessary to achieve the desired goal of histological MRI.
- Publication type
- Journal Article MeSH
Greater wax moth (GWM), Galleria mellonella (Lepidoptera: Pyralidae), is a highly destructive honey bee pest prevalent throughout the world. It is considered as a major factor to the alarming decline in honey bee population. GWM destroys active honey combs as it feeds on the beeswax and lays eggs in bee hives, and the primary food of their larva is beeswax. Beeswax is a polymer composed mainly of saturated and unsaturated, linear and complex monoesters, and hydrocarbons. The most frequent bond in beeswax is ethene (CH2-CH2) which is also found in the common plastic polyethylene. As wax-digestion is not a common animal character, we hypothesized about a possible role of GWM gut microflora in the process; which could possibly degrade polyethylene-like polymers as well. This study was aimed to identify the GWM gut microflora via culture-dependant approach. We characterized several bacterial species based on the culture characteristics, Gram-reaction, and various biochemical tests. Sequencing of 16S-rDNA revealed nine bacterial and one microalgal species from GWM gut. The bacterial species included Gram-positive Exiguobacterium aestuarii, Bacillus circulans, Microbacterium zaea, Microbacterium sp. and Enterococcus faecalis; Gram-negative Agrobacterium sp., Sphingomonas pseudosanguinis, Sphingobium yanoikuyae and Acinetobacter radioresistens; the microalgae was Picochlorum oklahomensis. Some of them have been previously reported to degrade polycyclic aromatic hydrocarbon, low-density polyethylene, and 2-methylphenanthrene. Meanwhile, the microalga, P. oklahomensis, was reported to steal bacterial genes to adapt with abiotic stresses. Further investigation is necessary to explore the precise details about polymer degrading capabilities of these microbes; nevertheless, this study builds a foundation for elaborate and advanced future research.
BACKGROUND: ADNP is essential for embryonic development. As such, de novo ADNP mutations lead to an intractable autism/intellectual disability syndrome requiring investigation. METHODS: Mimicking humans, CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 editing produced mice carrying heterozygous Adnp p.Tyr718∗ (Tyr), a paralog of the most common ADNP syndrome mutation. Phenotypic rescue was validated by treatment with the microtubule/autophagy-protective ADNP fragment NAPVSIPQ (NAP). RESULTS: RNA sequencing of spleens, representing a peripheral biomarker source, revealed Tyr-specific sex differences (e.g., cell cycle), accentuated in females (with significant effects on antigen processing and cellular senescence) and corrected by NAP. Differentially expressed, NAP-correctable transcripts, including the autophagy and microbiome resilience-linked FOXO3, were also deregulated in human patient-derived ADNP-mutated lymphoblastoid cells. There were also Tyr sex-specific microbiota signatures. Phenotypically, Tyr mice, similar to patients with ADNP syndrome, exhibited delayed development coupled with sex-dependent gait defects. Speech acquisition delays paralleled sex-specific mouse syntax abnormalities. Anatomically, dendritic spine densities/morphologies were decreased with NAP amelioration. These findings were replicated in the Adnp+/- mouse, including Foxo3 deregulation, required for dendritic spine formation. Grooming duration and nociception threshold (autistic traits) were significantly affected only in males. Early-onset tauopathy was accentuated in males (hippocampus and visual cortex), mimicking humans, and was paralleled by impaired visual evoked potentials and correction by acute NAP treatment. CONCLUSIONS: Tyr mice model ADNP syndrome pathology. The newly discovered ADNP/NAP target FOXO3 controls the autophagy initiator LC3 (microtubule-associated protein 1 light chain 3), with known ADNP binding to LC3 augmented by NAP, protecting against tauopathy. NAP amelioration attests to specificity, with potential for drug development targeting accessible biomarkers.
- MeSH
- Autistic Disorder * pathology MeSH
- Gene Expression MeSH
- Homeodomain Proteins genetics MeSH
- Humans MeSH
- Intellectual Disability * genetics metabolism MeSH
- Brain metabolism MeSH
- Mice MeSH
- Nerve Tissue Proteins genetics MeSH
- tau Proteins MeSH
- Tauopathies * metabolism MeSH
- Evoked Potentials, Visual MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
BACKGROUND: Variants of linker histone H1 are tissue-specific and are responsible for chromatin compaction accompanying cell differentiation, mitotic chromosome condensation, and apoptosis. Heterochromatinization, as the main feature of these processes, is also associated with pronounced trimethylation of histones H3 at the lysine 9 position (H3K9me3). METHODS: By confocal microscopy, we analyzed cell cycle-dependent levels and distribution of phosphorylated histone H1 (H1ph) and H3K9me3. By mass spectrometry, we studied post-translational modifications of linker histones. RESULTS: Phosphorylated histone H1, similarly to H3K9me3, has a comparable level in the G1, S, and G2 phases of the cell cycle. A high density of phosphorylated H1 was inside nucleoli of mouse embryonic stem cells (ESCs). H1ph was also abundant in prophase and prometaphase, while H1ph was absent in anaphase and telophase. H3K9me3 surrounded chromosomal DNA in telophase. This histone modification was barely detectable in the early phases of mitosis. Mass spectrometry revealed several ESC-specific phosphorylation sites of H1. HDAC1 depletion did not change H1 acetylation but potentiated phosphorylation of H1.2/H1.3 and H1.4 at serine 38 positions. CONCLUSIONS: Differences in the level and distribution of H1ph and H3K9me3 were revealed during mitotic phases. ESC-specific phosphorylation sites were identified in a linker histone.
- Publication type
- Journal Article MeSH
High-salt diets are a major cause of hypertension and cardiovascular (CV) disease. Many governments are interested in using food salt reduction programs to reduce the risk for salt-induced increases in blood pressure and CV events. It is assumed that reducing the salt concentration of processed foods will substantially reduce mean salt intake in the general population. However, contrary to expectations, reducing the sodium density of nearly all foods consumed in England by 21% had little or no effect on salt intake in the general population. This may be due to the fact that in England, as in other countries including the U.S.A., mean salt intake is already close to the lower normal physiologic limit for mean salt intake of free-living populations. Thus, mechanism-based strategies for preventing salt-induced increases in blood pressure that do not solely depend on reducing salt intake merit attention. It is now recognized that the initiation of salt-induced increases in blood pressure often involves a combination of normal increases in sodium balance, blood volume and cardiac output together with abnormal vascular resistance responses to increased salt intake. Therefore, preventing either the normal increases in sodium balance and cardiac output, or the abnormal vascular resistance responses to salt, can prevent salt-induced increases in blood pressure. Suboptimal nutrient intake is a common cause of the hemodynamic disturbances mediating salt-induced hypertension. Accordingly, efforts to identify and correct the nutrient deficiencies that promote salt sensitivity hold promise for decreasing population risk of salt-induced hypertension without requiring reductions in salt intake.
Oxidative stress can lead to various derivatives of the tyrosine residue in peptides and proteins. A typical product is 3-nitro-L-tyrosine residue (Nit), which can affect protein behavior during neurodegenerative processes, such as those associated with Alzheimer's and Parkinson's diseases. Surface enhanced Raman spectroscopy (SERS) is a technique with potential for detecting peptides and their metabolic products at very low concentrations. To explore the applicability to Nit, we use SERS to monitor tyrosine nitration in Met-Enkephalin, rev-Prion protein, and α-synuclein models. Useful nitration indicators were the intensity ratio of two tyrosine marker bands at 825 and 870 cm-1 and a bending vibration of the nitro group. During the SERS measurement, a conversion of nitrotyrosine to azobenzene containing peptides was observed. The interpretation of the spectra has been based on density functional theory (DFT) simulations. The CAM-B3LYP and ωB97XD functionals were found to be most suitable for modeling the measured data. The secondary structure of the α-synuclein models was monitored by electronic and vibrational circular dichroism (ECD and VCD) spectroscopies and modeled by molecular dynamics (MD) simulations. The results suggest that the nitration in these peptides has a limited effect on the secondary structure, but may trigger their aggregation.
- MeSH
- Azo Compounds chemistry MeSH
- Circular Dichroism MeSH
- Peptides chemical synthesis chemistry MeSH
- Spectrum Analysis, Raman methods MeSH
- Protein Structure, Secondary MeSH
- Molecular Dynamics Simulation MeSH
- Density Functional Theory MeSH
- Tyrosine analogs & derivatives analysis MeSH
- Publication type
- Journal Article MeSH
