One of the main challenges in analyzing chemical messengers in the brain is the optimization of tissue sampling and preparation protocols. Limiting postmortem time and terminating enzyme activity is critical to identify low-abundance neurotransmitters and neuropeptides. Here, we used a rapid and uniform conductive heat transfer stabilization method that was compared with a conventional fresh freezing protocol. Together with a selective chemical derivatization method and an optimized quantitation approach using deuterated internal standards, we spatially mapped neurotransmitters and their related metabolites by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) in rat brain tissue sections. Although the heat stabilization did not show differences in the levels of dopamine, norepinephrine, and serotonin, their related metabolites 3,4-dihydroxyphenylacetaldehyde, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 3-methoxy-4-hydroxyphenylacetaldehyde, dihydroxyphenylethyleneglycol, and 5-hydroxyindoleacetic acid were all significantly lower, indicating reduced neurotransmitter postmortem turnover ratios. Heat stabilization enabled detection of an increased number and higher levels of prodynorphin, proenkephalin, and tachykinin-derived bioactive neuropeptides. The low-abundant C-terminal flanking peptide, neuropeptide-γ, and nociceptin remained intact and were exclusively imaged in heat-stabilized brains. Without heat stabilization, degradation fragments of full-length peptides occurred in the fresh frozen tissues. The sample preparation protocols were furthermore tested on rat brains affected by acute anesthesia induced by isoflurane and medetomidine, showing comparable results to non-anesthetized animals on the neurotransmitters level without significant changes. Our data provide evidence for the potential use of heat stabilization prior to MALDI-MSI analyses to improve the examination of the in vivo state of neuronal chemical messengers in brain tissues not impacted by prior acute anesthesia.
- MeSH
- Rats MeSH
- Brain Chemistry * physiology MeSH
- Brain * metabolism MeSH
- Neurons * metabolism chemistry MeSH
- Neurotransmitter Agents * metabolism analysis MeSH
- Rats, Sprague-Dawley MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization * methods MeSH
- Hot Temperature * MeSH
- Freezing MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Current antibiotics and chemotherapeutics are becoming ineffective because pathogenic bacteria and tumor cells have developed multiple drug resistance. Therefore, it is necessary to find new substances that can be used in treatment, either alone or as sensitizing molecules in combination with existing drugs. Peptaibols are bioactive, membrane-active peptides of non-ribosomal origin, mainly produced by filamentous fungi such as Trichoderma spp. This study focused on producing peptaibol-rich extracts from Trichoderma atroviride O1, cultivated on malt extract agar (MA) under circadian and constant darkness conditions for 13 days. Peptaibol production was detected by MALDI-TOF mass spectrometry after six days of cultivation. The extracts demonstrated antibacterial activity against Staphylococcus aureus strains, particularly the methicillin-resistant variant, but not against the Gram-negative Pseudomonas aeruginosa. Quorum sensing interference revealed that a peptaibol-rich extract suppressed Vibrio campbellii BAA-1119's AI-2 signaling system to a degree comparable with gentamycin. Beyond antibacterial properties, the extracts exhibited notable antiproliferative activity against human ovarian cancer cells and their adriamycin-resistant subline in both 2D and 3D models. Specifically, MA-derived extracts reduced ovarian cancer cell viability by 70% at 50 μg/mL, especially under light/dark regime of cultivation. Compared to previously published results for PDA-based extracts, MA cultivation shifted the biological effects of peptaibol-containing extracts toward anticancer potential. These findings support the idea that modifying fungal cultivation parameters, the bioactivity of secondary metabolite mixtures can be tailored for specific therapeutic applications.
- MeSH
- Agar * chemistry MeSH
- Anti-Bacterial Agents * pharmacology metabolism MeSH
- Hypocreales MeSH
- Culture Media chemistry MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Cell Line, Tumor MeSH
- Peptaibols * pharmacology metabolism biosynthesis chemistry MeSH
- Cell Proliferation drug effects MeSH
- Antineoplastic Agents * pharmacology metabolism MeSH
- Pseudomonas aeruginosa drug effects MeSH
- Staphylococcus aureus drug effects MeSH
- Trichoderma * metabolism growth & development chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Skin represents the largest organ in the human body, functioning as a protective barrier against environmental factors while playing a critical role in thermoregulation. Acne vulgaris is recognized as the most common dermatological condition affecting adolescents, and if left untreated, it can result in lasting skin damage and associated psychosocial challenges. This study aims to develop innovative polymeric biomaterials that could effectively support the treatment of acne vulgaris. The synthesis of these biomaterials involves the use of polyethylene glycol 6000, sodium alginate, and the antioxidant protein glutathione (GHS) to create polymeric hydrogels. These hydrogels were generated via a UV-mediated crosslinking process. To enhance the functional properties of the hydrogels, zinc oxide microparticles (ZnO), synthesized through a wet precipitation method, were incorporated into the formulations. Characterization of the ZnO was performed using Fourier-Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), particle sizer analysis, and Scanning Electron Microscopy (SEM). Additionally, the bioactivity of the synthesized materials was evaluated through incubation in media simulating physiological body fluids. The cytotoxic effects of the biomaterials were assessed using an indirect test on mouse fibroblast (L929) cells, in accordance with ISO 10993-5 guidelines. The results of our research indicate that the developed biomaterials exhibit potential as a carrier for active substances, contributing positively to the treatment of acne vulgaris and potentially improving overall skin health.
- MeSH
- Acne Vulgaris drug therapy MeSH
- Alginates chemistry MeSH
- Biocompatible Materials chemistry pharmacology MeSH
- Cell Line MeSH
- Fibroblasts drug effects metabolism MeSH
- Glutathione * metabolism MeSH
- Hydrogels * chemistry MeSH
- Skin * drug effects metabolism MeSH
- Humans MeSH
- Mice MeSH
- Drug Carriers chemistry MeSH
- Zinc Oxide * chemistry pharmacology MeSH
- Regeneration drug effects MeSH
- Spectroscopy, Fourier Transform Infrared MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Zázvor lékařský (Zingiber officinale) je vytrvalá bylina původem z jihovýchodní Asie. V tradiční medicíně se využívá již tisíce let k léčbě mnoha onemocnění, např. zažívacích potíží, diabetu, infekcí a neurologických onemocnění. Používá se také jako koření. Oddenek zázvoru obsahuje velké množství bioaktivních látek – silice, flavonoidy, fenolické kyseliny a další. Některé léčivé účinky zázvoru již potvrdily i klinické studie (antidiabetický, antiemetický). Zázvor působí také na nervový systém – anxiolyticky, antidepresivně, neuroprotektivně, prokognitivně a antikonvulzivně. Je důležité zdůraznit, že zázvor lékařský není běžně používán jako lék v psychiatrii, avšak existuje několik studií a průzkumů, které naznačují některé potenciální benefity zázvoru v oblasti psychického zdraví. Tato oblast výzkumu však dosud stále probíhá a následující informace jsou tedy spíše hypotetické a vyžadují další výzkum pro potvrzení.
Ginger (Zingiber officinale) is a perennial herb native to Southeast Asia. It has been used in traditional medicine for thousands of years to treat many diseases, such as digestive problems, diabetes, infections, and neurological diseases. It is also used as a spice. The rhizome of ginger contains many bioactive substances – essential oils, flavonoids, phenolic acids, and others. Clinical studies have confirmed some medicinal effects of ginger (antidiabetic, antiemetic). Ginger also affects the nervous system – anxiolytic, antidepressant, neuroprotective, procognitive, and anticonvulsant. It is important to point out that ginger is not commonly used as a medicine in psychiatry. Still, several studies and research suggest some potential mental health benefits of ginger. However, this area of research is still ongoing, and the accompanying information is therefore rather hypothetical and requires further research for confirmation.
Polyphenolic compounds play an essential role in plant growth, reproduction, and defense mechanisms against pathogens and environmental stresses. Extracting these compounds is the initial step in assessing phytochemical changes, where the choice of extraction method significantly influences the extracted analytes. However, due to environmental factors, analyzing numerous samples is necessary for statistically significant results, often leading to the use of harmful organic solvents for extraction. Therefore, in this study, a novel DES-based shaking-assisted extraction procedure for the separation of polyphenolic compounds from plant samples followed by LC-ESI-QTOF-MS analysis was developed. The DES was prepared from choline chloride (ChCl) as the hydrogen bond acceptor (HBA) and fructose (Fru) as the hydrogen bond donor (HBD) at various molar ratios with the addition of 30% water to reduce viscosity. Several experimental variables affecting extraction efficiency were studied and optimized using one-variable-at-a-time (OVAT) and confirmed by response surface design (RS). Nearly the same experimental conditions were obtained using both optimization methods and were set as follows: 30 mg of sample, 300 mg of ChCl:Fru 1:2 DES containing 30% w/w of water, 500 rpm shaking speed, 30 min extraction time, 10°C extraction temperature. The results were compared with those obtained using conventional solvents, such as ethanol, methanol and water, whereby the DES-based shaking-assisted extraction method showed a higher efficiency than the classical procedures. The greenness of the developed method was compared with the greenness of existing procedures for the extraction of polyphenolic substances from solid plant samples using the complementary green analytical procedure index (ComplexGAPI) approach, while the results for the developed method were better or comparable to the existing ones. In addition, the practicability of the developed procedure was evaluated by application of the blue applicability grade index (BAGI) metric. The developed procedure was applied to the determination of spruce root samples with satisfactory results and has the potential for use in the analysis of similar plant samples.
- Publication type
- Journal Article MeSH
Solanum viarum, a perennial shrub, belongs to the family Solanaceae known for its therapeutic value worldwide. As a beneficial remedial plant, it is used for treating several disorders like dysentery, diabetes, inflammation, and respiratory disorders. Phytochemistry studies of this plant have shown the presence of steroidal glycoside alkaloids, including solasonine, solasodine, and solamargine. It also has flavonoids, saponins, minerals, and other substances. S. viarum extracts and compounds possess a variety of pharmacological effects, including antipyretic, antioxidant, antibacterial, insecticidal, analgesic, and anticancer activity. Most of the heavy metals accumulate in the aerial sections of the plant which is considered a potential phytoremediation, a highly effective method for the treatment of metal-polluted soils. We emphasize the forgoing outline of S. viarum, as well as its ethnomedicinal and ethnopharmacological applications, the chemistry of its secondary metabolites, and heavy metal toxicity. In addition to describing the antitumor activity of compounds and their mechanisms of action isolated from S. viarum, liabilities are also explained and illustrated, including any significant chemical or metabolic stability and toxicity risks. A comprehensive list of information was compiled from Science Direct, PubMed, Google Scholar, and Web of Science using different key phrases (traditional use, ethnomedicinal plants, western Himalaya, Himachal Pradesh, S viarum, and biological activity). According to the findings of this study, we hope that this review will inspire further studies along the drug discovery pathway of the chemicals extracted from the plant of S. viarum. Further, this review shows that ethnopharmacological information from ethnomedicinal plants can be a promising approach to drug discovery for cancer and diabetes.
- MeSH
- Biodegradation, Environmental * MeSH
- Ethnopharmacology * MeSH
- Phytochemicals pharmacology isolation & purification MeSH
- Phytotherapy MeSH
- Humans MeSH
- Plant Extracts * pharmacology chemistry isolation & purification MeSH
- Solanum * chemistry MeSH
- Medicine, Traditional * methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Cardiovascular diseases (CVDs) are a group of diseases with a very high rate of morbidity and mortality. The clinical presentation of CVDs can vary from asymptomatic to classic symptoms such as chest pain in patients with myocardial infarction. Current therapeutics for CVDs mainly target disease symptoms. The most common CVDs are coronary artery disease, acute myocardial infarction, atrial fibrillation, chronic heart failure, arterial hypertension, and valvular heart disease. In their treatment, conventional therapies and pharmacological therapies are used. However, the use of herbal medicines in the therapy of these diseases has also been reported in the literature, resulting in a need for critical evaluation of advances related to their use. Therefore, we carried out a narrative review of pharmacological and herbal therapeutic effects reported for these diseases. Data for this comprehensive review were obtained from electronic databases such as MedLine, PubMed, Web of Science, Scopus, and Google Scholar. Conventional therapy requires an individual approach to the patients, as when patients do not respond well, this often causes allergic effects or various other unwanted effects. Nowadays, medicinal plants as therapeutics are frequently used in different parts of the world. Preclinical/clinical pharmacology studies have confirmed that some bioactive compounds may have beneficial therapeutic effects in some common CVDs. The natural products analyzed in this review are promising phytochemicals for adjuvant and complementary drug candidates in CVDs pharmacotherapy, and some of them have already been approved by the FDA. There are insufficient clinical studies to compare the effectiveness of natural products compared to approved therapeutics for the treatment of CVDs. Further long-term studies are needed to accelerate the potential of using natural products for these diseases. Despite this undoubted beneficence on CVDs, there are no strong breakthroughs supporting the implementation of natural products in clinical practice. Nevertheless, they are promising agents in the supplementation and co-therapy of CVDs.
- Publication type
- Journal Article MeSH
- Review MeSH
Černucha setá (Nigella sativa L.) je jednoletá bylina z čeledi pryskyřníkovitých, původní v západní Asii a severní Africe. Její semena se používají jako koření, zejména v Indii a na Středním východě, ale také k léčebným účelům. Semena černuchy obsahují velké množství bioaktivních látek, které vykazují množství farmakologických účinků. Černucha setá patří mezi donedávna zapomenuté léčivky a v různých systémech tradiční medicíny se používá již tisíce let, ale studie moderní medicíny posledních let ukazují, že její léčebné využití může být mnohem širší. Je pravděpodobné, že by mohla najít využití i v léčbě psychických poruch, jako jsou úzkost, deprese a některá neurodegenerativní onemocnění. Bez zajímavosti není ani její potenciální využití v terapii covidu-19.
Nigella sativa is an annual herb of the buttercup family, native to Western Asia and North Africa. Its seeds are used as a spice, especially in India and the Middle East, but also for medicinal purposes. Nigella sativa seeds contain a large number of bioactive substances, which have a number of pharmacological effects. It is a forgotten medicine and has been used in various systems of traditional medicine for thousands of years, but studies of modern medicine in recent years show that its therapeutic use can be much wider. It is likely that it could also find use in the treatment of mental disorders such as anxiety, depression and some neurodegenerative diseases. Its potential use in COVID-19 therapy is not without interest.
- MeSH
- Dementia MeSH
- Mental Disorders MeSH
- Phytotherapy MeSH
- Rodentia MeSH
- Clinical Studies as Topic MeSH
- Humans MeSH
- Disease Models, Animal MeSH
- Nervous System * drug effects MeSH
- Nigella sativa * chemistry drug effects MeSH
- Plant Preparations pharmacology therapeutic use MeSH
- Seizures MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Review MeSH
In the realm of surgical and dental applications, hyaluronic acid (HA) braided threads show significant therapeutic potential due to their incorporation of pharmaceutical active ingredients. This study primarily focuses on resolving the crucial challenge of devising a deposition method that can ensure both precision and uniformity in the content of the active ingredient Octenidine dihydrochloride (OCT) within each segment of the threads. Our objective in this study was to develop a continuous deposition method for OCT onto a braided thread composed of 24 hyaluronic acid-based fibers, aiming for a specific OCT content of 0.125 μg/mm, while maintaining a maximum allowable deviation of ±15% in OCT content. The motivation behind designing this novel method stemmed from the necessity of employing a volatile solvent for the active agent. Conventional wetting methods proved unsuitable due to fluctuations in the solution's concentration during deposition, and alternative methods known to us demanded intricate technical implementations. The newly introduced method offers distinct advantages, including its online processing speed, scalability potential, and cost-efficiency of the active agent solution. Additionally, it minimizes the impact on the natural polymer thread, preserving energy by obviating the need for complete thread saturation. Our research and precise apparatus development resulted in achieving the desired thread properties, with an OCT content of (1.51 ± 0.09) μg per 12 mm thread piece. These findings not only validate the suitability of this innovative method for depositing active agents but also extend its potential applicability beyond dental care.
- Publication type
- Journal Article MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
nestr.
Léčba diabetických ran je problematická, časově i finančně náročná a omezující pro pacienty. Krevní destičky jsou fyziologicky přítomné v procesu hojení akutní rány, kde produkují růstové a angiogenní faktory, cytokiny, chemokiny, molekuly zprostředkující adhesi buněk a další bioaktivní látky napomáhající hojení. Lyzát připravený z plazmy obohacené o krevní destičky bude zabudován do degradovatelných i nedegradovatelných nanovlákenných nosičů připravených ze syntetických i přirozených polymerů dále modifikovaných fibrinem, a to s cílem postupného uvolňování růstových faktorů a dalších biologicky aktivních látek. Bioaktivita těchto nanovlákenných krytů bude hodnocena in vitro v kulturách lidských keratinocytů, dermálních fibroblastů, cévních endotelových buněk a kmenových buněk tukové tkáně. Nanovlákenné kryty s nejlepšími vlastnostmi budou aplikovány in vivo na rány u diabetických potkanů, a následně bude hodnocena jejich schopnost stimulace hojení ran.; Treatment of diabetic wounds is problematic, long-lasting, expensive and restrictive for patients. Platelets are physiologically present in the process of healing of acute wounds, where they secret growth and angiogenic factors, cytokines, chemokines, cell adhesion-mediating molecules and other bioactive substances that support healing. Lysate prepared from platelet-rich plasma will be incorporated into degradable and non-degradable nanofibrous dressings, prepared from synthetic and natural polymers further modified with fibrin, in order to assure controlled release of growth factors and other bioactive substances. Bioactivity of these dressings will be evaluated in vitro in cultures of human keratinocytes, dermal fibroblasts, vascular endothelial cells and adipose tissue-derived stem cells. Nanofibrous dressings with the best properties will be applied in vivo on wounds of diabetic rats, and their ability to stimulate wound healing will be evaluated.
- Keywords
- řízené uvolňování, diabetické rány, lyzát z trombocytů, diabetický potkan, hojení ran, wound healing, diabetic wounds, platelet lysate, control release, diabetic rat, plasma obohacená destičkami, epidermální keratinocyty, dermální fibroblasty, kmenové buňky tukové tkáně, platelet-rich plasma, epidermal keratinocytes, dermal fibroblasts, adipose-tissue stem cells,
- NML Publication type
- závěrečné zprávy o řešení grantu AZV MZ ČR
