From algae to advancements: laminarin in biomedicine.

Laminarin, a complicated polysaccharide originating from brown algae, has emerged as a compelling candidate in the domain of biomedical research. This enigmatic molecule, composed of glucose units associated with both ß-1,3 and ß-1,6 glycosidic bonds, possesses an array of remarkable characteristics that render it auspicious for multifaceted biomedical applications. This review investigates the comprehensive potential of laminarin in the biomedical domain, emphasizing its remarkable biocompatibility, low cytotoxicity, and cell proliferation support. Laminarin's immunomodulatory attributes position it as an encouraging contender in immunotherapy and the development of vaccines. Moreover, its anti-inflammatory and antioxidant characteristics provide a promising avenue for combatting conditions associated with oxidative stress. In particular, laminarin excels as a drug delivery vehicle owing to its exceptional encapsulation capabilities emerging from its porous framework. Integrating pH and redox responsiveness in laminarin-based drug delivery systems is poised to redefine targeted therapies. Laminarin substantially contributes to tissue engineering by improving adhesion, migration of cells, and deposition of extracellular matrix. This augmentation magnifies the regenerative capability of tissue-engineered constructs, substantiated by the advancement of laminarin-based wound dressings and tissue scaffolds, marking considerable progress in the domain of wound healing and tissue regeneration. While laminarin exhibits substantial potential in biomedical applications, it remains in the initial phases of exploration. Comprehensive preclinical and clinical research is warranted to verify its effectiveness and safety across various applications. In essence, laminarin, a marine marvel, has the capability to remodel biomedical research, offering inventive solutions to complex difficulties.

Structural and biochemical characterisation of the N-carbamoyl-ß-alanine amidohydrolase from Rhizobium radiobacter MDC 8606.

N-carbamoyl-ß-alanine amidohydrolase (CßAA) constitutes one of the most important groups of industrially relevant enzymes used in the production of optically pure amino acids and derivatives. In this study, a CßAA-encoding gene from Rhizobium radiobacter strain MDC 8606 was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme (RrCßAA) showed a specific activity of 14 U·mg-1 using N-carbamoyl-ß-alanine as a substrate with an optimum activity at 55 °C and pH 8.0. In this work, we report also the first prokaryotic CßAA structure at a resolution of 2.0 Å. A discontinuous catalytic domain and a dimerisation domain attached through a flexible hinge region at the domain interface have been revealed. We identify key ligand binding residues, including a conserved glutamic acid (Glu131), histidine (H385) and arginine (Arg291). Our results allowed us to explain the preference of the enzyme for linear carbamoyl substrates, as large and branched carbamoyl substrates cannot fit in the active site of the enzyme. This work envisages the use of RrCßAA from R. radiobacter MDC 8606 for the industrial production of L-α-, L-ß- and L-γ-amino acids. The structural analysis provides new insights on enzyme-substrate interaction, which shed light on engineering of CßAAs for high catalytic activity and broad substrate specificity.

Peptide-based targeted cancer therapeutics: Design, synthesis and biological evaluation.

Cancer is the leading cause for human mortality together with cardiovascular diseases. Abl (Abelson) tyrosine kinases play a fundamental role in transducing various signals that control proliferation, survival, migration and invasion in several cancers such as Chronic Myeloid Leukemia (CML), breast cancer and brain cancer. For these reasons Abl tyrosine kinases are considered important biological targets in drug discovery. In this study a series of lysine-based oligopeptides with expected Abl inhibitory activity were designed resembling the binding of FDA-approved drugs (i.e. of Imatinib and Nilotinib), synthesized, purified by High Performance Liquid Chromatography (HPLC), analyzed by mass spectrometry (MS) and biologically tested in vitro in CML (AR-230 and K-562), breast cancers (MDA-MB 231 and MDA-MB 468) and glioblastoma cell lines (U87 and U118). The solid-phase peptide synthesis (SPPS) by Fmoc (9-fluorenylmethoxycarbonyl) chemistry was used to synthesize target compounds. AutoDock Vina was applied for simulation binding to Abl. The biological activities were measured evaluating cytotoxic effect, induction of apoptosis and inhibition of cancer cells migration. The new peptides exhibited different concentration-dependent antiproliferative effect against the tumor cell lines after 72 h treatment. The most promising results were obtained with the U87 glioblastoma cell line where a significant reduction of the migration ability was detected with one compound (H-Lys1-Lys2-Lys3-NH2).

Proof of the feasibility of a nanocell-based wide-range optical magnetometer.

We present an experimental scheme performing scalar magnetometry based on the fitting of Rb $ {{\rm D}_2} $D2 line spectra recorded by derivative selective reflection spectroscopy from an optical nanometric-thick cell. To demonstrate its efficiency, the magnetometer is used to measure the inhomogeneous magnetic field produced by a permanent neodymium--iron-boron alloy ring magnet at different distances. The computational tasks are realized by relatively cheap electronic components: an Arduino Due board for external control of the laser and acquisition of spectra, and a Raspberry Pi computer for the fitting. The coefficient of variation of the measurements remains under 5% in the magnetic field range of 40-200 mT, limited only by the size of the oven and translation stage used in our experiment. The proposed scheme is expected to operate with high measurement precision also for stronger magnetic fields ($ {\gt} {500}\;{\rm mT}$>500mT) in the hyperfine Paschen-Back regime, where the evolution of atomic transitions can be calculated with high accuracy.

Analysis of genome sequence and trehalose lipid production peculiarities of the thermotolerant Gordonia strain.

Gordoniae are one of the most promising hydrocarbon-oxidizing actinobacteria. Here we present the genome sequence analysis of thermotolerant strain Gordonia sp. 1D isolated from oil-refinery soil. It is capable of alkane consumption and biosurfactant production at temperatures of up to 50°C. Gordonia sp. 1D demonstrates maximum biosurfactant production when grown on hexadecane, and at 40°C it was slightly higher than at 27°C: 35 and 39 mN/m, respectively. For the first time, it was experimentally confirmed that the carbohydrate component of extracellular biosurfactants produced by strain 1D is trehalose. In addition, genes for the production of trehalose lipid biosurfactants were identified. The genetic determinants for two different pathways for trehalose synthesis were found. The strain carries genes otsA and otsB involved in de novo trehalose biosynthesis. Moreover, the genes treY and treZ responsible for trehalose biosynthesis from maltooligosaccharides and starch or glycogen were identified.

A universal automated tool for reliable detection of seizures in rodent models of acquired and genetic epilepsy.

OBJECTIVE: Prolonged electroencephalographic (EEG) monitoring in chronic epilepsy rodent models has become an important tool in preclinical drug development of new therapies, in particular those for antiepileptogenesis, disease modification, and treating drug-resistant epilepsy. We have developed an easy-to-use, reliable, computational tool for automated detection of electrographic seizures from prolonged EEG recordings in rodent models of epilepsy. METHODS: We applied a novel method based on advanced time-frequency analysis that detects EEG episodes with excessive activity in certain frequency bands. The method uses an innovative technique of short-term spectral analysis, the Similar Basis Function algorithm. The method was applied for offline seizure detection from long-term EEG recordings from four spontaneously seizing, chronic epilepsy rat models: the fluid percussion injury (n = 5 rats, n = 49 seizures) and post-status epilepticus models (n = 119 rats, n = 993 seizures) of acquired epilepsy, and two genetic models of absence epilepsy, Genetic Absence Epilepsy Rats from Strasbourg and Wistar Albino Glaxo from Rijswijk (n = 41 and 14 rats, n = 8733 and 825 seizures, respectively). RESULTS: Our comparative analysis revealed that the EEG amplitude spectra of these four rat models are remarkably similar during epileptiform activity and have a single expressed peak within the 17- to 25-Hz frequency range. Focusing on this band, our computer program detected all seizures in the 179 rats. A quick semiautomated user inspection of the EEGs for the period of each identified event allowed quick rejection of artifact events. The overall processing time for 12-day-long recordings varied from a few minutes (5-10) to 30 minutes, depending on the number of artifact events, which was strongly correlated with the signal quality of the raw EEG data. SIGNIFICANCE: Our automated seizure detection tool provides high sensitivity, with acceptable specificity, for long- and short-term EEG recordings from both acquired and genetic chronic epilepsy rat models. This tool has the potential to improve the efficiency and rigor of preclinical research and therapy development using these models.

Dark resonance formation with magnetically induced transitions: extension of spectral range and giant circular dichroism.

Dark resonances were formed via electromagnetically induced transparency for the first time, to the best of our knowledge, involving magnetically induced ΔF=±2 atomic transitions of alkali metal atoms, which are forbidden at zero magnetic field. The probability of these transitions undergoes rapid growth when 300-3000 G magnetic field is applied, allowing formation of dark resonances, widely tunable in the GHz range. It is established that for ΔF=+2 (ΔF=-2) transition, the coupling laser tuned to ΔF=+1 (ΔF=-1) transition of the hyperfine Λ-system must be σ+ (σ-) polarized, manifesting anomalous circular dichroism.

Selective reflection from an Rb layer with a thickness below λ/12 and applications.

We have studied the peculiarities of selective reflection from an Rb vapor cell with a thickness L<70 nm, which is smaller than the length scale of evanescent fields λ/2π and more than an order of magnitude smaller than the optical wavelength. A 240 MHz redshift due to the atom-surface interaction is observed for a cell thickness of L=40 nm. In addition, complete frequency-resolved hyperfine Paschen-Back splitting of atomic transitions to four components for Rb87 and six components for Rb87 is recorded in a strong magnetic field (B>2 kG).

N-type resonances in a buffered micrometric Rb cell: splitting in a strong magnetic field.

N-type resonances excited in rubidium atoms confined in micrometric-thin cells with variable thickness from 1 µm to 2 mm are studied experimentally for the cases of a pure Rb atomic vapor and of a vapor with neon buffer gas. Good contrast and narrow linewidth were obtained for thicknesses as low as 30 µm. The higher amplitude and sharper profile of N-type resonances in the case of a buffered cell was exploited to study the splitting of the 85Rb D1 N-resonance in a magnetic field of up to 2200 G. The results are fully consistent with the theory. The mechanism responsible for forming N-resonances is discussed. Possible applications are addressed.

Three-photon electromagnetically induced transparency using Rydberg states.

We demonstrate electromagnetically induced transparency in a four-level cascade system where the upper level is a Rydberg state. The observed spectral features are sub-Doppler and can be enhanced due to the compensation of Doppler shifts with AC Stark shifts. A theoretical description of the system is developed that agrees well with the experimental results, and an expression for the optimum parameters is derived.

Hyperfine Paschen-Back regime realized in Rb nanocell.

A simple and efficient scheme based on a one-dimensional nanometric-thin cell filled with Rb and strong permanent ring magnets allows direct observation of the hyperfine Paschen-Back regime on the D(1) line in the 0.5-0.7 T magnetic field. Experimental results are perfectly consistent with the theory. In particular, with σ(+) laser excitation, the slopes of the B-field dependence of frequency shifts for all 10 individual transitions of (85,87)Rb are the same and equal to 18.6 MHz/mT. Possible applications for magnetometry with submicron spatial resolution and tunable atomic frequency references are discussed.

Rhythmogram-based analysis for continuous electrographic data of the human brain.

Ecologically relevant stimuli are rarely used in scientific studies because they are difficult to control. Instead, researchers employ simple stimuli with sharp boundaries (in space and time). Here, we explore how the rhythmogram can be used to provide much needed rigorous control of natural continuous stimuli like music and speech. The analysis correlates important features in the time course of stimuli with corresponding features in brain activations elicited by the same stimuli. Correlating the identified regularities of the stimulus time course with the features extracted from the activations of each voxel of a tomographic analysis of brain activity provides a powerful view of how different brain regions are influenced by the stimulus at different times and over different (user-selected) timescales. The application of the analysis to tomographic solutions extracted from magnetoencephalographic data recorded while subjects listen to music reveals a surprising and aesthetically pleasing aspect of brain function: an area believed to be specialized for visual processing is recruited to analyze the music after the acoustic signal is transformed to a feature map. The methodology is ideal for exploring processing of complex stimuli, e.g., linguistic structure and meaning and how it fails, for example, in developmental dyslexia.

Essential features of optical processes in neon-buffered submicron-thin Rb vapor cell.

A new submicron thin cell (STC) filled with Rb and neon gas is developed and comparison of resonant absorption with STC containing pure Rb is provided. The effect of collapse and revival of Dicke-type narrowing is still observable for the thickness L = lambda /2 and L = lambda , where lambda is a resonant laser wavelength 794 nm (D(1) line). For an ordinary Rb cm-size cell with addition of buffer gas, the velocity selective optical pumping/saturation (VSOP) resonances in saturated absorption spectra are fully suppressed if neon pressure > 0.5 Torr. A spectacular difference is that for L = lambda , VSOP resonances are still observable even when neon pressure is > or = 6 Torr. Narrow fluorescence spectra at L = lambda /2 allow one to realize online buffer gas pressure monitoring. A good agreement with theoretical model is observed.

Simulation of sleep spindles and spike and wave discharges using a novel method for the calculation of field potentials in rats.

We suggest a new method for calculation of extracellular field potentials generated by a large population of pyramidal cells (PCs), using a single PC compartmental model. Similar methods described earlier use the assumption that the intracellular potential or current distributions of the cells within the population are much alike as a result of simultaneous activation at about the same longitudinal location (i.e., all the PCs in the population are located on the same level and are ideally synchronized). However, the degree of synchronization of natural firing even during synchronized rhythmic discharges in the cortex is not as high. We introduce the possibility to vary the degree of synchronization of the PCs' activity in the population, thus taking into account disperse timing of cortical pyramidal cells' firing. The temporal variability in cell firing is described by a Gaussian distribution, the width of which defines the degree of synchronization/desynchronization. In addition, the suggested method allows for certain spatial spread of PCs in the population along longitudinal axis of the PCs. The method was applied to test the assumption that the transition from sleep spindles to rhythmic spike and wave discharges (SWDs) observed in absence epilepsy may occur due to an increase in pyramidal cells' firing synchronization. We show that in case of weak synchronization of PC firing in the population, the shape of field potential during rhythmic thalamic input is similar to the oscillations during a sleep spindle, while at stronger synchronization of PCs, it looks much more as a SWD, with clear expressed spikes and waves. This suggests that in large population of pyramidal cells the changes in the degree of synchronization of cell firing may explain the changes in the shape of field potential from spindle oscillations to SWDs and vice versa.

A computer model of field potential responses for the study of short-term plasticity in hippocampus.

Activity-dependent synaptic plasticity has important implications for network function. The previously developed model of the hippocampal CA1 area, which contained pyramidal cells (PC) and two types of interneurons involved in feed-forward and recurrent inhibition, respectively, and received synaptic inputs from CA3 neurons via the Schaffer collaterals, was enhanced by incorporating dynamic synaptic connections capable of changing their weights depending on presynaptic activation history. The model output was presented as field potentials, which were compared with those derived experimentally. The parameters of Schaffer collateral-PC excitatory model synapse were determined, with which the model successfully reproduced the complicated dynamics of train-stimulation sequential potentiation/depression observed in experimentally recorded field responses. It was found that the model better reproduces the time course of experimental field potentials if the inhibitory synapses on PC are also made dynamic, with expressed properties of frequency-dependent depression. This finding supports experimental evidence that these synapses are subject to activity-dependent depression. The model field potentials in response to various randomly generated and real (derived from recorded CA3 unit activity) long stimulating trains were calculated, illustrating that short-term plasticity with the observed characteristics could play specific roles in frequency processing in hippocampus and thus providing a new tool for the theoretical study of activity-dependent synaptic plasticity.

A model synapse that incorporates the properties of short- and long-term synaptic plasticity.

We propose a general computer model of a synapse, which incorporates mechanisms responsible for the realization of both short- and long-term synaptic plasticity-the two forms of experimentally observed plasticity that seem to be very significant for the performance of neuronal networks. The model consists of a presynaptic part based on the earlier 'double barrier synapse' model, and a postsynaptic compartment which is connected to the presynaptic terminal via a feedback, the sign and magnitude of which depend on postsynaptic Ca(2+) concentration. The feedback increases or decreases the amount of neurotransmitter which is in a ready for release state. The model adequately reproduced the phenomena of short- and long-term plasticity observed experimentally in hippocampal slices for CA3-CA1 synapses. The proposed model may be used in the investigation of certain real synapses to estimate their physiological parameters, and in the construction of realistic neuronal networks.