Accumulation of environmental chitin in the lungs can lead to pulmonary fibrosis, characterized by inflammatory infiltration and fibrosis in acidic chitinase (Chia)-deficient mice. Transgenic expression of Chia in these mice ameliorated the symptoms, indicating the potential of enzyme supplementation as a promising therapeutic strategy for related lung diseases. This study focuses on utilizing hyperactivated human Chia, which exhibits low activity. We achieved significant activation of human Chia by incorporating nine amino acids derived from the crab-eating monkey (Macaca fascicularis) Chia, known for its robust chitin-degrading activity. The modified human Chia retained high activity across a broad pH spectrum and exhibited enhanced thermal stability. The amino acid substitutions associated with hyperactivation of human Chia activity occurred species specifically in monkey Chia. This discovery highlights the potential of hyperactivated Chia in treating pulmonary diseases resulting from chitin accumulation in human lungs.

• MeSH
• Enzyme Activation drug effects   MeSH
• Chitin metabolism   chemistry   MeSH
• Chitinases * metabolism   genetics   chemistry   MeSH
• Hydrogen-Ion Concentration MeSH
• Humans MeSH
• Macaca fascicularis MeSH
• Mice MeSH
• Lung metabolism   pathology   enzymology   MeSH
• Enzyme Stability MeSH
• Amino Acid Substitution MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Microbial transglutaminase (MTG) is an enzyme widely used in the food industry because it creates cross-links between proteins, enhancing the texture and stability of food products. Its unique properties make it a valuable tool for modifying the functional characteristics of proteins, significantly impacting the quality and innovation of food products. In this study, response surface methodology was employed to optimize the fermentation conditions for microbial transglutaminase production by the strain Streptoverticillium cinnamoneum KKP 1658. The effects of nitrogen dose, cultivation time, and initial pH on the activity of the produced transglutaminase were investigated. The significance of the examined factors was determined as follows: cultivation time > nitrogen dose > pH. The interaction between nitrogen dose and cultivation time was found to be crucial, having the second most significant impact on transglutaminase activity. Optimal conditions were identified as 48 h of cultivation with a 2% nitrogen source dose and an initial medium pH of approximately 6.0. Under these conditions, transglutaminase activity ranged from 4.5 to 5.5 U/mL. The results of this study demonstrated that response surface methodology is a promising approach for optimizing microbial transglutaminase production. Future applications of transglutaminase include the development of modern food products with improved texture and nutritional value, as well as its potential use in regenerative medicine for creating biomaterials and tissue scaffolds. This topic is particularly important and timely as it addresses the growing demand for innovative and sustainable solutions in the food and biomedical industries, contributing to an improved quality of life.

• MeSH
• Nitrogen * metabolism   MeSH
• Fermentation * MeSH
• Hydrogen-Ion Concentration MeSH
• Culture Media * chemistry   MeSH
• Transglutaminases * metabolism   MeSH
• Publication type
• Journal Article MeSH

The present study has undertaken the isolation of marine yeasts from mangrove sediment samples and their ability to produce alkaline protease enzymes. A total of 14 yeast isolates were recovered on yeast-malt agar (YMA) and yeast extract peptone dextrose (YEPD) agar medium. After screening for proteolytic activity on skim milk agar, marine yeast isolate, AKB-1 exhibited a hydrolysis zone of 18 mm. Optimal conditions for the enzyme production from yeast isolate AKB-1 were at 30 °C, pH 8, fructose as carbon source, potassium nitrate as nitrogen source, and 25% saline concentration. Under the optimal conditions, the protease enzyme activity of the isolate AKB-1 was observed to be 978 IU/mL. The structural and functional analysis was carried out through FTIR and HPLC analysis for the extracted protease enzyme. Furthermore, the enzyme produced was partially purified by solvent extraction using ethyl acetate and ammonium sulfate precipitation (3.4-fold) followed by dialysis (56.8-fold). The molecular weight of the purified enzyme was observed to be around 60 kDa using SDS-PAGE. The extracted protein showed good antibacterial activity against six different clinical bacterial pathogens and the highest against Bacillus cereus (16 ± 0.5 mm). The extracted protease enzyme was revealed to remove blood stains from cloth within 20 min of application similar to the commercial detergent. The marine yeast isolate was further identified as Candida orthopsilosis AKB-1 (Accession number KY348766) through 18S rRNA sequencing, and a phylogenetic tree was generated.

• MeSH
• Anti-Bacterial Agents pharmacology   metabolism   chemistry   isolation & purification   MeSH
• Bacillus cereus drug effects   MeSH
• Bacterial Proteins * chemistry   pharmacology   metabolism   isolation & purification   MeSH
• Candida * enzymology   isolation & purification   genetics   classification   MeSH
• Endopeptidases * chemistry   metabolism   isolation & purification   pharmacology   MeSH
• Phylogeny MeSH
• Geologic Sediments microbiology   MeSH
• Hydrogen-Ion Concentration MeSH
• Culture Media chemistry   MeSH
• Microbial Sensitivity Tests MeSH
• Molecular Weight MeSH
• Enzyme Stability MeSH
• Temperature MeSH
• Publication type
• Journal Article MeSH

β-Glucans comprise a group of β-D-glucose polysaccharides (glucans) that occur naturally in the cell walls of bacteria, fungi, and cereals. Its degradation is catalyzed by β-glucanases, enzymes that catalyze the breakdown of β-glucan into cello-oligosaccharides and glucose. These enzymes are classified as endo-glucanases, exo-glucanases, and glucosidases according to their mechanism of action, being the lichenases (β-1,3;1,4-glucanases, EC 3.2.1.73) one of them. Hence, we aimed to enhance lichenase production by Thermothelomyces thermophilus through the application of response surface methodology, using tamarind (Tamarindus indica) and jatoba (Hymenaea courbaril) seeds as carbon sources. The crude extract was immobilized, with a focus on improving lichenase activity, using various ionic supports, including MANAE (monoamine-N-aminoethyl), DEAE (diethylaminoethyl)-cellulose, CM (carboxymethyl)-cellulose, and PEI (polyethyleneimine)-agarose. Regarding lichenase, the optimal conditions yielding the highest activity were determined as 1.5% tamarind seeds, cultivation at 50 °C under static conditions for 72 h. Moreover, transitioning from Erlenmeyer flasks to a bioreactor proved pivotal, resulting in a 2.21-fold increase in activity. Biochemical characterization revealed an optimum temperature of 50 °C and pH of 6.5. However, sustained stability at varying pH and temperature levels was challenging, underscoring the necessity of immobilizing lichenase on ionic supports. Notably, CM-cellulose emerged as the most effective immobilization medium, exhibiting an activity of 1.01 U/g of the derivative (enzyme plus support), marking a substantial enhancement. This study marks the first lichenase immobilization on these chemical supports in existing literature.

• MeSH
• Enzymes, Immobilized * metabolism   chemistry   MeSH
• Fungal Proteins * metabolism   chemistry   MeSH
• Glycoside Hydrolases * metabolism   chemistry   biosynthesis   MeSH
• Hydrogen-Ion Concentration MeSH
• Fruit metabolism   MeSH
• Seeds metabolism   MeSH
• Sordariales MeSH
• Enzyme Stability MeSH
• Tamarindus metabolism   microbiology   MeSH
• Publication type
• Journal Article MeSH

Feruloyl esterases (FAEs) are a crucial component of the hemicellulose-degrading enzyme family that facilitates the degradation of lignocellulose while releasing hydroxycinnamic acids such as ferulic acid with high added value. Currently, the low enzyme yield of FAEs is one of the primary factors limiting its application. Therefore, in this paper, we optimized the fermentation conditions for the expression of FAE BpFaeT132C-D143C with excellent thermal stability in Escherichia coli by experimental design. Firstly, we explored the effects of 11 factors such as medium type, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration, and inoculum size on BpFaeT132C-D143C activity separately by the single factor design. Then, the significance of the effects of seven factors, such as post-induction temperature, shaker rotational speed, and inoculum size on BpFaeT132C-D143C activity, was analyzed by Plackett-Burman design. We identified the main factors affecting the fermentation conditions of E. coli expressing BpFaeT132C-D143C as post-induction temperature, pre-induction period, and post-induction period. Finally, we used the steepest ascent path design and response surface method to optimize the levels of these three factors further. Under the optimal conditions, the activity of BpFaeT132C-D143C was 3.58 U/ml, which was a significant 6.6-fold increase compared to the pre-optimization (0.47 U/ml), demonstrating the effectiveness of this optimization process. Moreover, BpFaeT132C-D143C activity was 1.52 U/ml in a 3-l fermenter under the abovementioned optimal conditions. It was determined that the expression of BpFaeT132C-D143C in E. coli was predominantly intracellular in the cytoplasm. This study lays the foundation for further research on BpFaeT132C-D143C in degrading agricultural waste transformation applications.

• MeSH
• Escherichia coli * genetics   metabolism   enzymology   MeSH
• Fermentation * MeSH
• Isopropyl Thiogalactoside metabolism   MeSH
• Carboxylic Ester Hydrolases * genetics   metabolism   chemistry   biosynthesis   MeSH
• Culture Media chemistry   MeSH
• Coumaric Acids metabolism   MeSH
• Lignin MeSH
• Recombinant Proteins genetics   metabolism   biosynthesis   chemistry   MeSH
• Enzyme Stability MeSH
• Temperature MeSH
• Publication type
• Journal Article MeSH

Tento článek shrnuje roli telomer a enzymu telomerázy v reprodukční fyziologii zvířat a možnosti jejich terapeutického ovlivnění. Telomery, nacházející se na koncích chromozomů, hrají klíčovou roli v ochraně genomové stability a buněčné proliferace. Jejich zkracování může negativně ovlivnit fertilitu, kvalitu gamet a embryonální vývoj. Telomeráza je enzym, který prodlužuje telomery a je aktivní především v kmenových a zárodečných buňkách. Poznatky z humánní medicíny naznačují, že podpora její aktivity může mít pozitivní vliv na repro dukční úspěšnost samců i samic. Protože biologie telomer je u lidí i zvířat velmi podobná, lze poznatky přenést do veterinární medicíny (Fick, 2012). Tento článek analyzuje dostupné studie a zkoumá potenciální využití telomerázové terapie ve veterinární praxi.

This article summarizes the role of telomeres and the enzyme telomerase in animal reproductive physiology and explores potential the rapeutic interventions. Telomeres, located at the ends of chromosomes, play a crucial role in maintaining genomic stability and cellular proliferation. Their shortening can negatively impact fertility, gamete quality, and embryonic development. Telomerase is an enzyme that extends telomeres and is primarily active in stem and germ cells. Findings from human medicine suggest that enhancing its activity may positively influence reproductive success in both males and females. Since telomere biology is highly similar between humans and animals, these insights can be translated into veterinary medicine (Fick, 2012). This article reviews current studies and examines the potential applications of telomerase therapy in veterinary practice.

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

Purine de novo purine synthesis involves 10 reactions catalysed by six enzymes, including phosphoribosylformyglycinamidine synthase (PFAS). To date, genetic defects of three of these enzymes, namely ATIC, ADSL and PAICS, have been characterised in humans. Here, we report for the first time two individuals with PFAS deficiency. Probands were identified through metabolic and genetic screening of neurologically impaired individuals. The pathogenicity of the variants was established by structural and functional studies. Probands C1 and C2 presented with prematurity, short stature, recurrent seizures and mild neurological impairment. C1 had elevated urinary levels of formylglycineamide riboside (FGAr) and bi-allelic PFAS variants encoding the NP_036525.1:p.Arg811Trp substitution and the NP_036525.1:p.Glu228_Ser230 in-frame deletion. C2 is a 20-year-old female with a homozygous NP_036525.1:p.Asn264Lys substitution. These amino acid changes are predicted to affect the structural stability of PFAS. Accordingly, C1 skin fibroblasts showed decreased PFAS content and activity, with impaired purinosome formation that was restored by transfection with pTagBFP_PFAS_wt. The enzymatic activities of the corresponding recombinant mutant PFAS proteins were also reduced, and none of them, after transfection, corrected the elevated FGAR/r levels in PFAS-deficient HeLa cells. While genetic defects in purine de novo synthesis are typically considered in patients with severe neurological impairment, these disorders, especially PFAS deficiency, should also be considered in milder phenotypes.

• MeSH
• Humans MeSH
• Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor * genetics   deficiency   metabolism   MeSH
• Young Adult MeSH
• Mutation MeSH
• Purine-Pyrimidine Metabolism, Inborn Errors * genetics   MeSH
• Child, Preschool MeSH
• Purines * biosynthesis   MeSH
• Check Tag
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Case Reports MeSH

Vysokoškolská příručka a učebnice, která se zaměřuje na farmaceutickou technologii, konkrétně na disoluční metody. Určeno i odborníkům v praxi.

• MeSH
• Pharmacokinetics MeSH
• Pharmaceutical Preparations MeSH
• Dosage Forms MeSH
• Drug Compounding MeSH
• Solubility MeSH
• Drug Development MeSH
• Publication type
• Handbook MeSH

• Conspectus
• Farmacie. Farmakologie
• NML Fields
• farmacie a farmakologie
• NML Publication type
• učebnice vysokých škol

Alveolární echinokokóza (AE) je vzácné, ale závažné parazitární onemocnění jater, které často napodobuje maligní ložiska. V této kazuistice prezentujeme případ 76letého pacienta, u něhož byla AE diagnostikována náhodně při vyšetření pro renální insuficienci. Ultrazvukové vyšetření odhalilo mnohočetná hyperechogenní ložiska v játrech. Následné CT a MR jater zobrazily mnohočetná ložiska nepravidelných okrajů bez typické vaskularizace, největší o velikosti 71 × 38 mm. Laboratorní výsledky ukázaly zvýšené jaterní enzymy, CRP a renální insuficienci. Core cut biopsie jater potvrdila přítomnost pseudocystických struktur. Vzorky byly následně zaslány do Národní referenční laboratoře pro tkáňové helmintózy, která diagnózu AE potvrdila. Sérologické vyšetření bylo provedeno až po bioptickém vyšetření a potvrdilo přítomnost protilátek proti Echinococcus multilocularis. Pacient byl odeslán do infekční ambulance, kde byla zahájena léčba albendazolem v dávce 800 mg denně. Po rehydrataci a úpravě léčby došlo ke stabilizaci renálních funkcí a při kontrolních zobrazovacích vyšetřeních byl nález stacionární. Tento případ zdůrazňuje nutnost zařazení AE do diferenciální diagnostiky ložiskových procesů jater, zejména u pacientů bez onkologické anamnézy.

Alveolar echinococcosis (AE) is a rare but serious parasitic liver disease that often mimics malignant lesions. We present the case of a 76-year-old patient in whom AE was incidentally diagnosed during examination for renal insufficiency. Abdominal ultrasound revealed multiple hyperechogenic liver lesions. Subsequent MRI showed multiple irregularly bordered lesions without typical vascularization, the largest measuring 71 × 38 mm. Laboratory tests revealed elevated liver enzymes, CRP, and renal insufficiency. A core-cut liver biopsy confirmed the presence of pseudocystic structures. Samples were sent to the National Reference Laboratory for Tissue Helminthiases, which confirmed the diagnosis of AE. Serological testing was performed after the biopsy confirming the presence of antibodies against Echinococcus multilocularis. The patient was referred to an infectious disease clinic, where treatment with albendazole at a dose of 800 mg daily was initiated. After rehydration and adjustment of therapy, renal function stabilized, and follow-up imaging showed stable findings. This case highlights the need to include AE in the differential diagnosis of hepatic lesions, particularly in patients without an oncological history.

• MeSH
• Albendazole pharmacology   therapeutic use   MeSH
• Biopsy MeSH
• Diagnostic Imaging methods   MeSH
• Echinococcosis, Hepatic * diagnosis   drug therapy   MeSH
• Liver diagnostic imaging   pathology   MeSH
• Humans MeSH
• Aged MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Aged MeSH
• Publication type
• Case Reports MeSH