The precise measurement of cell temperature and an in-depth understanding of thermogenic processes are critical in unraveling the complexities of cellular metabolism and its implications for health and disease. This review focuses on the mechanisms of local temperature generation within cells and the array of methods developed for accurate temperature assessment. The contact and noncontact techniques are introduced, including infrared thermography, fluorescence thermometry, and other innovative approaches to localized temperature measurement. The role of thermogenesis in cellular metabolism, highlighting the integral function of temperature regulation in cellular processes, environmental adaptation, and the implications of thermogenic dysregulation in diseases such as metabolic disorders and cancer are further discussed. The challenges and limitations in this field are critically analyzed while technological advancements and future directions are proposed to overcome these barriers. This review aims to provide a consolidated resource for current methodologies, stimulate discussion on the limitations and challenges, and inspire future innovations in the study of cellular thermodynamics.
- MeSH
- lidé MeSH
- teplota MeSH
- termogeneze * fyziologie MeSH
- termografie * metody MeSH
- termometrie metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy 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
- antibakteriální látky farmakologie metabolismus chemie izolace a purifikace MeSH
- Bacillus cereus účinky léků MeSH
- bakteriální proteiny * chemie farmakologie metabolismus izolace a purifikace MeSH
- Candida * enzymologie izolace a purifikace genetika klasifikace MeSH
- endopeptidasy * chemie metabolismus izolace a purifikace farmakologie MeSH
- fylogeneze MeSH
- geologické sedimenty mikrobiologie MeSH
- koncentrace vodíkových iontů MeSH
- kultivační média chemie MeSH
- mikrobiální testy citlivosti MeSH
- molekulová hmotnost MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- časopisecké články 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 * genetika metabolismus enzymologie MeSH
- fermentace * MeSH
- isopropylthiogalaktosid metabolismus MeSH
- karboxylesterhydrolasy * genetika metabolismus chemie biosyntéza MeSH
- kultivační média chemie MeSH
- kyseliny kumarové metabolismus MeSH
- lignin MeSH
- rekombinantní proteiny genetika metabolismus biosyntéza chemie MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
Druhé vydání 131 stran : ilustrace ; 23 cm
Publikace se zaměřuje na otužování, na fyziologické aspekty adaptace člověka na chlad. Určeno odborné i široké veřejnosti.; Monografie pojednává o různých aspektech reakce a adaptace lidského organismu na působení chladu se zvláštním zřetelem na chlad extrémní. Tomu jsou vystaveni sportovci (maratonští plavci, horolezci, vodáci, triatlonisté, sportovní otužilci a často také lyžaři), v profesní oblasti pak potápěči, vodní záchranáři, členové horské služby, vojáci, ale i všichni, kteří jsou nuceni pracovat v drsných klimatických podmínkách. Adaptace na chlad se týká samozřejmě i běžné, nesportující populace - otužování vůči chladu je důležité zejména z hlediska snížení nemocnosti pro akutní respirační choroby. Kniha podává přehled fyziologických pochodů nastupujících jako reakce na celkové působení intenzivního chladového podnětu, zabývá se projevy adaptace na chlad a objasňuje podstatu zvyšování obranyschopnosti organismu otužováním. Text je určen převážně lékařům, ale též pracovníkům v profesích vyžadujících adaptaci na chlad. Dále pak pedagogům a trenérům sportovců, kteří přicházejí s chladem do styku. Kompletní průvodce adaptací na zimu a chladné prostředí pro lékaře, sportovce i širokou veřejnost nabízí odpovědi na tyto otázky: Které fyziologické procesy stojí za adaptací na chlad? Jak funguje lidské tělo v extrémních podmínkách? Je otužování jen módní vlna, nebo má smysl pro všechny? Jak správně trénovat tělo na chlad a jak tyto poznatky aplikovat ve sportu i v každodenním životě? Jaká rizika jsou spojena s chladem - a jak jim předejít?
- Klíčová slova
- otužování,
- MeSH
- fyziologická adaptace MeSH
- nízká teplota MeSH
- sporty MeSH
- zdravé chování MeSH
- změny tělesné teploty MeSH
- Publikační typ
- monografie MeSH
- populární práce MeSH
- Konspekt
- Fyziologie člověka a srovnávací fyziologie
- NLK Obory
- fyziologie
- zdravotní výchova
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
- krysa rodu rattus MeSH
- mozek - chemie * fyziologie MeSH
- mozek * metabolismus MeSH
- neurony * metabolismus chemie MeSH
- neurotransmiterové látky * metabolismus analýza MeSH
- potkani Sprague-Dawley MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice * metody MeSH
- vysoká teplota * MeSH
- zmrazování MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In the study, we employ an affordable, tissue-saving, and precise simultaneous multiplex immunofluorescence method with heat-induced antibody stripping to identify and structurally analyse nigral Lewy bodies in dopaminergic neurones. Analysis of different alpha-synuclein epitopes and proteoforms reveals an almost uniform, onion-like morphology of the Lewy bodies. The N-terminal and C-terminal domains are predominantly accessible to antibody binding in the peripheral shell of the bodies.
- MeSH
- alfa-synuklein * metabolismus analýza MeSH
- demence s Lewyho tělísky patologie MeSH
- dopaminergní neurony metabolismus patologie MeSH
- fluorescenční protilátková technika metody MeSH
- Lewyho tělíska * metabolismus patologie MeSH
- lidé MeSH
- substantia nigra * metabolismus patologie MeSH
- vysoká teplota MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Azo dyes are used as coloring agent in textile industries at larger scale. As a result, large quantity of dye-enriched waste water is generated which subsequently poses environmental problems. Biological tool involving bacteria having azoreductase enzyme has proved to be more effective and efficient in dye effluent treatment. Current work focuses on Staphylococcus caprae (S. caprae) for degradation and decolorization of Reactive Red-195 (RR-195) azo dye. For this purpose, factors such as pH, temperature, inoculums, carbon and nitrogen sources, and dye concentrations have been optimized for maximum decolorization and degradation. S. caprae (4 mg/mL) efficiently resulted into 90% decolorization of RR-195 dye under static condition at 100 μg/mL concentration, 30 °C and pH 7.0 at a 12-h contact period. FTIR analysis has revealed the formation of new functional groups in the treated dye such as O-H stretch at 3370 cm-1, C-H band stretching at 2928 cm-1, and new band at 1608 cm-1 which specify the degradation of aromatic ring, 1382 and 1118 cm-1 represents desulfonated peaks. Biodegraded metabolites of RR-195 dye such as phenol, 3, 5-di-tert-butylphenol, and phthalic acid have been identified respectively that find industrial applications. Phytotoxicity test has shown non-toxic effects of treated dye on germination of Vigna radiata and Triticum aestivum seeds. Further, antibiotic diffusion assay has confirmed the biosafety of S. caprae.
- MeSH
- azosloučeniny * metabolismus toxicita MeSH
- barvicí látky * metabolismus MeSH
- biodegradace * MeSH
- chemické látky znečišťující vodu metabolismus MeSH
- dusík metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- odpadní voda * mikrobiologie chemie MeSH
- průmyslový odpad MeSH
- Staphylococcus capitis metabolismus izolace a purifikace MeSH
- Staphylococcus metabolismus MeSH
- teplota MeSH
- textilie MeSH
- textilní průmysl MeSH
- uhlík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Human milk harbors diverse bacterial communities that contribute to infant health. Although pumping and storing milk is a common practice, the viable bacterial composition of pumped milk and the impact of storage practice on these bacteria remains under-explored. This metagenomic observational study aimed to characterize viable bacterial communities in freshly pumped human milk and its changes under different storage conditions. METHODS: In 2023, twelve lactating mothers from the CELSPAC: TNG cohort (Czech Republic) provided freshly pumped milk samples. These samples were stored under various conditions (refrigeration for 24 h, 48 h, or freezing for six weeks) and treated with propidium monoazide (PMA) to selectively identify viable cells. The DNA extracted from individual samples was subsequently analyzed using 16S rRNA amplicon sequencing on the Illumina platform. RESULTS: The genera Streptococcus, Staphylococcus, Diaphorobacter, Cutibacterium, and Corynebacterium were the most common viable bacteria in fresh human milk. The median sequencing depth and Shannon index of fresh human milk samples treated with PMA (+ PMA) were significantly lower than in untreated (-PMA) samples (p < 0.05 for all), which was true also for each time point. Also, significant changes in these parameters were observed between fresh human milk samples and their paired frozen samples (p < 0.05), while no differences were found between fresh human milk samples and those refrigerated for up to 48 h (p > 0.05). Of specific genera, only + PMA frozen human milk samples showed a significant decrease in the central log-ratio transformed relative abundances of the genera Diaphorobacter and Cutibacterium (p < 0.05) in comparison to + PMA fresh human milk samples. CONCLUSIONS: The study demonstrated that the bacterial profiles significantly differed between human milk samples treated with PMA, which represent only viable bacteria, and those untreated. While storage at 4 °C for up to 48 h did not significantly alter the overall diversity and composition of viable bacteria in human milk, freezing notably affected both the viability and relative abundances of some bacterial genera.
- MeSH
- azidy MeSH
- Bacteria * izolace a purifikace genetika klasifikace MeSH
- chlazení MeSH
- dospělí MeSH
- lidé MeSH
- mateřské mléko * mikrobiologie MeSH
- mikrobiota * MeSH
- propidium analogy a deriváty MeSH
- RNA ribozomální 16S MeSH
- skladování potravin * metody MeSH
- zmrazování MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
145 stran : ilustrace ; 30 cm
Směrnice, které se zaměřují na proces sterilizace zdravotnických prostředků pomocí páry a tepla. Určeno odborné veřejnosti.
- MeSH
- pára MeSH
- reprodukovatelnost výsledků MeSH
- řízení kvality MeSH
- sektor zdravotní péče MeSH
- sterilizace MeSH
- vysoká teplota MeSH
- zdravotnické prostředky MeSH
- Publikační typ
- směrnice MeSH
- Geografické názvy
- Česká republika MeSH
- Evropa MeSH
- Konspekt
- Metrologie. Standardizace
- NLK Obory
- technika lékařská, zdravotnický materiál a protetika
It has been demonstrated that freezing-induced acidity changes have an impact on the structural integrity, degree of aggregation, and chemical stability of frozen food and pharmaceutical products. The stability of the compounds in solutions is maintained by the presence of buffers. However, many buffers are unsuitable for applications involving freezing as this process substantially alters the acidity. In this study, we determine the effect of initial pH, concentration, and cooling rate on the freezing-induced change in acidity of phosphate buffered saline (PBS) in the frozen state via UV-VIS spectroscopy. Furthermore, we examine the impact of individual salts present in PBS and discuss the mechanisms affecting the resulting acidity that we approximate via Hammett acidity function (H2-).
- MeSH
- fosfáty * chemie MeSH
- koncentrace vodíkových iontů MeSH
- pufry MeSH
- solný roztok * chemie MeSH
- zmrazování MeSH
- Publikační typ
- časopisecké články MeSH