Hyaluronan and hyaluronidases are critical in tissue remodeling, inflammation, and tumor progression. This chapter provides a comprehensive guide to hyaluronan zymography, a powerful technique for detecting and quantifying hyaluronidase activity in complex biological samples. The method involves separating proteins by polyacrylamide gel electrophoresis with hyaluronan incorporated into the gel matrix. Following electrophoresis, the gel is incubated to allow hyaluronidases to degrade the hyaluronan substrate, resulting in clear digestion zones. Detailed protocols for sample preparation and the zymographic process are included, offering researchers a robust tool for studying hyaluronidase activity and regulation in various biological contexts.
In the last two decades, a school of thought emerged that perceives male reproductive health, testicular function, and sperm output as a sentry for general, somatic health. Large-scale epidemiologic studies have already linked the reduced sperm count to increased risk of chronic somatic disease (e.g., cancer, cardiovascular, neurological and bone diseases), yet most of these studies have not taken full advantage of advanced andrological analysis. Altered proteostasis, i.e., the disbalance between protein synthesis and turnover, is a common denominator of many diseases, including but not limited to cancer and neurodegenerative diseases. This chapter introduces the concept of cellular proteostasis as a measure of sperm structural and functional integrity and an endpoint of varied impacts on spermiogenesis and sperm maturation, including heritability, general health, lifestyle, and occupational and environmental reprotoxic exposure. Special consideration is given to small molecule protein modifiers, sperm-binding seminal plasma proteins, zinc-interacting proteins, and redox proteins responsible for the maintenance of protein structure and the protection of spermatozoa from oxidative damage. While the main focus is on human male infertility, serious consideration is given to relevant animal models, and in particular to male food animals with extensive records of fertility from artificial insemination services. Altogether, the proteostatic biomarker discovery and validation studies set the stage for the integration of proteomics of sperm proteostasis with genomic and high throughput phenomic approaches to benefit both human and animal reproductive medicine.
- MeSH
- fertilita * fyziologie MeSH
- homeostáze proteinů * fyziologie MeSH
- lidé MeSH
- mužská infertilita * metabolismus genetika patologie patofyziologie MeSH
- spermatogeneze * MeSH
- spermie * metabolismus patologie fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Cellular processes such as tissue regeneration, inflammation, and migration require the proteolysis of the extracellular matrix and the proteolytic activation of signaling molecules. A widely used and accessible technique for studying these processes is gelatin zymography, particularly for investigating matrix metalloproteinases (MMPs), though it is not limited to them. This method is favored for its simplicity, low cost, and robustness. Despite certain limitations, it remains a preferred approach for the initial investigation of complex samples.Here, we present a protocol applicable to various sample sources, including proteases from human cell lines and bacteria isolated from chronic wounds. We also explore changes in protease activity within exudates from human chronic wounds, a challenging analysis for more complex techniques. Additionally, we emphasize the potential to extend the basic protocol to study the conditions under which proteases are active.
BACKGROUND: Age-related neurodegenerative diseases (NDs) pose a formidable challenge to healthcare systems worldwide due to their complex pathogenesis, significant morbidity, and mortality. Scope and Approach: This comprehensive review aims to elucidate the central role of the microbiotagut- brain axis (MGBA) in ND pathogenesis. Specifically, it delves into the perturbations within the gut microbiota and its metabolomic landscape, as well as the structural and functional transformations of the gastrointestinal and blood-brain barrier interfaces in ND patients. Additionally, it provides a comprehensive overview of the recent advancements in medicinal and dietary interventions tailored to modulate the MGBA for ND therapy. CONCLUSION: Accumulating evidence underscores the pivotal role of the gut microbiota in ND pathogenesis through the MGBA. Dysbiosis of the gut microbiota and associated metabolites instigate structural modifications and augmented permeability of both the gastrointestinal barrier and the blood-brain barrier (BBB). These alterations facilitate the transit of microbial molecules from the gut to the brain via neural, endocrine, and immune pathways, potentially contributing to the etiology of NDs. Numerous investigational strategies, encompassing prebiotic and probiotic interventions, pharmaceutical trials, and dietary adaptations, are actively explored to harness the microbiota for ND treatment. This work endeavors to enhance our comprehension of the intricate mechanisms underpinning ND pathogenesis, offering valuable insights for the development of innovative therapeutic modalities targeting these debilitating disorders.
- MeSH
- dysbióza metabolismus MeSH
- hematoencefalická bariéra metabolismus MeSH
- lidé MeSH
- mozek * metabolismus MeSH
- neurodegenerativní nemoci * mikrobiologie metabolismus MeSH
- osa mozek-střevo * fyziologie MeSH
- probiotika MeSH
- stárnutí * MeSH
- střevní mikroflóra * fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Učební texty Univerzity Karlovy
3., upravené vydání 113 stran : ilustrace ; 30 cm
Vysokoškolská učebnice, která se zaměřuje na cytologii a obecnou histologii.
- MeSH
- cytologie MeSH
- histologie MeSH
- Konspekt
- Buněčná biologie. Cytologie
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- cytologie, klinická cytologie
- histologie
- NLK Publikační typ
- učebnice vysokých škol
2. vydání 143 stran : ilustrace (převážně barevné) ; 24 cm
Vysokoškolská příručka a učebnice, která se zaměřuje na farmaceutickou technologii, konkrétně na disoluční metody. Určeno i odborníkům v praxi.
- MeSH
- farmakokinetika MeSH
- léčivé přípravky MeSH
- lékové formy MeSH
- příprava léků MeSH
- rozpustnost MeSH
- vyvíjení léků MeSH
- Publikační typ
- příručky MeSH
- Konspekt
- Farmacie. Farmakologie
- NLK Obory
- farmacie a farmakologie
- NLK Publikační typ
- učebnice vysokých škol
This report presents a fatal case of a young female Type I diabetic patient who developed convulsions and loss of consciousness after taking methamphetamine and spending some time in a dance club. During the convulsions, she was given sugar and when no response occurred, her boyfriend who was not experienced in the use of insulin administered a dose of insulin to her. The woman lost consciousness and died despite the efforts of the emergency service. A biochemical analysis revealed a high level of insulin (196.67 mU/L) and low levels of glucose (2.96 mmol/L) and C-peptide (26 pmol/L). Toxicological analysis revealed a methamphetamine concentration of 389 ng/mL and an amphetamine concentration of 19 ng/mL. The forensic perspective of the difficult determination of the contribution of each of the factors to the death, i.e., the pre-existing medical condition (Type I diabetes), the use of methamphetamine, the physical exertion at the dance club, and, finally, the non-indicated administration of insulin, is discussed. The ruling of the court is also reported.
- MeSH
- bezvědomí chemicky indukované MeSH
- C-peptid krev MeSH
- diabetes mellitus 1. typu * MeSH
- dospělí MeSH
- fatální výsledek MeSH
- hypoglykemika škodlivé účinky MeSH
- inzulin * aplikace a dávkování MeSH
- krevní glukóza analýza MeSH
- lidé MeSH
- methamfetamin * škodlivé účinky MeSH
- poruchy spojené s užíváním amfetaminu komplikace MeSH
- stimulanty centrálního nervového systému * škodlivé účinky MeSH
- tanec MeSH
- tělesná námaha MeSH
- záchvaty MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
Rare and unknown actinobacteria from unexplored environments have the potential to produce new bioactive molecules. This study aimed to use 16 s rRNA metabarcoding to determine the composition of the actinobacterial community, particularly focusing on rare and undescribed species, in a nature reserve within the Brazilian Cerrado called Sete Cidades National Park. Since this is an inaccessible area without due legal authorization, it is understudied, and, therefore, its diversity and biotechnological potential are not yet fully understood, and it may harbor species with groundbreaking genetic potential. In total, 543 operational taxonomic units (OTUs) across 14 phyla were detected, with Actinobacteria (41.2%), Proteobacteria (26.5%), and Acidobacteria (14.3%) being the most abundant. Within Actinobacteria, 107 OTUs were found, primarily from the families Mycobacteriaceae, Pseudonocardiaceae, and Streptomycetaceae. Mycobacterium and Streptomyces were the predominant genera across all samples. Seventeen rare OTUs with relative abundance < 0.1% were identified, with 82.3% found in only one sample yet 25.5% detected in all units. Notable rare and transient genera included Salinibacterium, Nocardia, Actinomycetospora_01, Saccharopolyspora, Sporichthya, and Nonomuraea. The high diversity and distribution of Actinobacteria OTUs indicate the area's potential for discovering new rare species. Intensified prospection on underexplored environments and characterization of their actinobacterial diversity could lead to the discovery of new species capable of generating innovative natural products.
- MeSH
- Actinobacteria * chemie klasifikace genetika izolace a purifikace MeSH
- biodiverzita MeSH
- metagenom MeSH
- půda chemie MeSH
- půdní mikrobiologie * MeSH
- RNA ribozomální 16S analýza MeSH
- taxonomické DNA čárové kódování MeSH
- veřejné parky MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Brazílie MeSH
- MeSH
- diabetes mellitus 2. typu * diagnóza MeSH
- lidé MeSH
- selfmonitoring glykemie metody MeSH
- Check Tag
- lidé MeSH
