Intensive fish farming is associated with a high level of stress, causing immunosuppression. Immunomodulators of natural origin, such as probiotics or phytoadditives, represent a promising alternative for increasing the immune function of fish. In this study, we tested the autochthonous trout probiotic strain L. plantarum R2 in a newly developed, low-cost application form ensuring the rapid revitalization of bacteria. We tested continuous and cyclic feeding regimes with regard to their effect on the intestinal immune response and microbiota of rainbow trout. We found that during the continuous application of probiotic feed, the immune system adapts to the immunomodulator and there is no substantial stimulation of the intestinal immune response. During the cyclic treatment, after a 3-week break in probiotic feeding and the reintroduction of probiotics, there was a significant stimulation of the gene expression of molecules associated with both cellular and humoral immunity (CD8, TGF-β, IL8, TLR9), without affecting the gene expression for IL1 and TNF-α. We can conclude that, in aquaculture, this probiotic feed can be used with a continuous application, which does not cause excessive immunostimulation, or with a cyclic application, which provides the opportunity to stimulate the immunity of trout, for example, in periods of stress.

• Klíčová slova
• gut microbiota, intestinal immune answer, probiotic feed, salmonid,
• Publikační typ
• časopisecké články MeSH

Disease conditions that involve multiple predisposing or contributing factors, or manifest as low performance and/or low-level mortality, can pose a diagnostic challenge that requires an interdisciplinary approach. Reaching a diagnosis may also be limited by a lack of available clinical profile parameter reference ranges to discriminate healthy fish from those affected by specific disease conditions. Here, we describe our experience investigating poorly performing rainbow trout (Oncorhynchus mykiss) in an intensive recirculation aquaculture, where reaching a final diagnosis of nephrocalcinosis was not as straightforward as one would wish. To list the issues making the diagnosis difficult, it was necessary to consider the creeping onset of the problem. Further diagnostic steps needed to ensure success included obtaining comparative data for fish blood profiles and water quality from both test and control aquacultural systems, excluding infections with salmonid pathogenic agents and evaluating necropsy findings. Major events in the pathophysiology of nephrocalcinosis could be reconstructed as follows: aquatic environment hyperoxia and hypercapnia → blood hypercapnia → blood acid-base perturbation (respiratory acidosis) → metabolic compensation (blood bicarbonate elevation and kidney phosphate excretion) → a rise in blood pH → calcium phosphate precipitation and deposition in tissues. This case highlights the need to consider the interplay between water quality and fish health when diagnosing fish diseases and reaching causal diagnoses.

• Klíčová slova
• acid-base, aquaculture, blood acid-base balance, carbon dioxide, fish health, rainbow trout, water quality,
• Publikační typ
• časopisecké články MeSH

The distribution of mercury species was studied in all aquatic ecosystem components (i.e., water, sediment, emergent aquatic plants, invertebrates and omnivorous and piscivorous fish) of the Záskalská water reservoir (Central Bohemia, Czech Republic) which is in the vicinity of an abandoned cinnabar mine. The results indicate that the transport of mercury from the cinnabar mine is the major source of mercury in the Záskalská reservoir. The legal maximum limit (0.07 μg/L) for total mercury concentration in water samples was exceeded only during rainy periods. The total mercury concentration in the surface sediments was in the range from 0.22 to 9.19 mg/kg in dry matter (up to 0.2% CH3Hg+) and was sample site-specific. The dominant form of mercury in sediments was mercury sulphide (22.9-79.2%). The emergent macrophytes accumulated mercury primarily by the roots from sediments, and no significant translocation of mercury to leaves was observed. The legal maximum limit for mercury content in fish muscle (0.5 mg/kg in the fresh matter) was exceeded up to 4.48 times for piscivorous fish. Hazard index values indicate a health risk concern for children and for people consuming more than 100 g of fish muscle per day. Our results emphasise the need to implement legal restrictions on the consumption of piscivorous fish caught in ecosystems downstream of abandoned cinnabar mines.

• Klíčová slova
• Aquatic ecosystem, Bioaccumulation, Hazard index, Mercury bioavailability, Mine waste,
• MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• ekosystém MeSH
• lidé MeSH
• monitorování životního prostředí MeSH
• rtuť * analýza   MeSH
• ryby MeSH
• sladká voda MeSH
• sloučeniny rtuti MeSH
• voda MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• cinnabar MeSH Prohlížeč
• rtuť * MeSH
• sloučeniny rtuti MeSH
• voda MeSH

Anastomotic leakage is a serious postoperative complication following a low anterior resection of rectum. Current research in colorectal surgery focuses on searching for techniques which could minimize the risk of leakage. The main objective of this study was to evaluate the impact of cumulative fluid balance on microcirculatory changes at the anastomotic site.This experimental study used 15 domestic pigs; all of the animals underwent rectal resection. Then the animals were divided into three groups and received IV crystalloids at various rates, i.e. 5 ml/kg/h; 10 ml/kg/h; and 20 ml/kg/h. Large bowel micro-perfusion was measured using laser Doppler flowmetry during and following the surgical procedure.The experiments were successfully performed in all 15 cases. No animal died during the procedure or during the follow-up. We found no differences in micro-perfusion of the bowel between the experimental groups during the surgical procedure and during the follow-up period after the surgery. A significant decrease in micro-perfusion was observed after the transection of the inferior mesenteric artery in all the groups; again, no differences between the groups were observed.We did not confirm the original hypothesis that excessive load of IV crystalloids during the surgery would have a negative effect on bowel micro-perfusion and thus a negative effect on the healing of the anastomosis. However, laser Doppler flowmetry was found to have high accuracy in measuring tissue microcirculation and has the potential to be used in clinical practice.

• Klíčová slova
• Anastomotic leakage, Blood flow, Laser Doppler flowmetry, Low anterior resection,
• Publikační typ
• časopisecké články MeSH

The aim of this study was to reveal the effects of foodborne fluoxetine on morphological and condition profile, hematological profile, biochemical and oxidative stress indices on juvenile rainbow trout. The study was performed according to OECD Guideline No. 215. Fluoxetine was incorporated into Biomar 921 pellets at a dose of 0.047 mg/kg (environmental concentration), 0.577 mg/kg and 6.7 mg/kg. There was statistically significant change in hematological profile, including an increasing trend in neutrophil/lymphocyte ratio and a decreasing trend in the number of lymphocytes. Measurements of oxidative stress indicated decreased activity of the detoxifying enzyme glutathione-S-transferase in the liver and kidney. However, the measurement of GR, GPx, CAT, SOD activity, and TBARS showed no changes. Histopathological examination revealed damage to the proximal tubules of caudal kidney in exposed groups. This study confirms that fluoxetine has a significant effect on immune response.

• Klíčová slova
• Aquatic pollutants, Fish hematological profile, Fish immune response, Oxidative stress, SSRI,
• MeSH
• antidepresiva druhé generace toxicita   MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• fluoxetin toxicita   MeSH
• imunita účinky léků   MeSH
• kontaminace potravin MeSH
• krevní obraz MeSH
• krmivo pro zvířata MeSH
• Oncorhynchus mykiss krev   imunologie   MeSH
• oxidační stres účinky léků   MeSH
• proximální tubuly ledvin účinky léků   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antidepresiva druhé generace MeSH
• chemické látky znečišťující vodu MeSH
• fluoxetin MeSH

INTRODUCTION: The inflammatory process in Crohn's disease (CD) is closely associated with the formation of reactive oxygen species. Antioxidant enzymes can play an important role in the outcome of CD and may influence postoperative recurrence in these patients. The aim of our study was to evaluate gene expression of intracellular antioxidant enzymes in surgically resected intestinal specimens of patients with CD, both in macroscopically normal and in inflamed tissue. METHODS: A total of 28 patients referred for elective bowel resection were enrolled in the study. Full-thickness small intestinal specimens were investigated. Gene expression of antioxidant enzymes - superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GSR) - was evaluated both in macroscopically normal and inflamed samples. RESULTS: There were significantly lower levels of SOD1 mRNA (p = 0.007) and GSR mRNA (p = 0.027) in inflamed tissue compared to macroscopically normal areas. No significant differences were found between affected and non-affected intestinal segments in mRNA for SOD2, SOD3 and GPX. CONCLUSIONS: Our pilot data clearly showed that the gene expression of major antioxidant enzymes is not a uniform mechanism in the pathogenesis of Crohn's disease. Topically decreased gene expression of SOD1 and GSR might facilitate the segmental tissue injury caused by reactive oxygen species.

• Klíčová slova
• Crohn’s disease, antioxidant enzymes, gene expression, glutathione peroxidase, glutathione reductase, intestine, superoxide dismutase,
• MeSH
• antioxidancia * MeSH
• Crohnova nemoc * genetika   metabolismus   MeSH
• exprese genu * MeSH
• glutathionperoxidasa genetika   MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• reaktivní formy kyslíku MeSH
• střeva MeSH
• superoxiddismutasa 1 * genetika   metabolismus   MeSH
• superoxiddismutasa genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antioxidancia * MeSH
• glutathionperoxidasa MeSH
• messenger RNA MeSH
• reaktivní formy kyslíku MeSH
• SOD1 protein, human MeSH Prohlížeč
• superoxiddismutasa 1 * MeSH
• superoxiddismutasa MeSH

BACKGROUND: The endothelial glycocalyx (EG) plays a vital role in the physiology and pathophysiology of human microcirculation. Having relevant EG damage model would be important tool for testing new interventions aiming at EG protection and recovery. We describe the first in vivo EG damage model in pig. OBJECTIVE: To investigate the course of animal EG damage induced by specific enzymes. MATERIAL AND METHODS: Four anesthetized piglets received enzymes: 1g hyaluronidase and 25 IU heparanase I intravenously. Blood and urine samples were collected at baseline and 20/40/60/80/100/120 min for detecting markers of endothelial and EG function. Sublingual microcirculation and EG thickness were assessed by Side-stream Dark Field (SDF) imaging and Perfused Boundary Region (PBR) respectively. EG of the mesentery artery was visualized in fluorescent microscopy. RESULTS: Biochemical marker of EG damage syndecan-1 showed temporary increase with return to baseline and was reflected by PBR values. Albumin levels suggested brief period of capillary leakage (decrease in the serum, increase in the urine) with a trend to normalization. Urine glycosaminoglycans peaked at 120 minutes. Microcirculatory perfusion parameter showed significant alteration. Diffusion parameters were altered with no statistical significance. CONCLUSION: EG damage induced by specific enzymes was reflected by temporary changes of biochemical makers together with alteration of microcirculation and changes in fluorescent microscopy of EG layer. Our results support to further validate presented model of EG damage on a larger number of animals.

• Klíčová slova
• Microcirculation, albuminuria, endothelial glycocalyx, heparanase, hyaluronidase,
• MeSH
• glykokalyx * MeSH
• kapiláry MeSH
• mikrocirkulace MeSH
• pilotní projekty MeSH
• prasata MeSH
• trávení MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The presence of circulating tumor cells (CTCs) in patients with solid tumors is associated with poor prognosis. However, there are limited data concerning the detection of CTCs in renal cell cancer (RCC). The aim of this study is to evaluate the presence of CTCs in peripheral blood of patients with RCC undergoing surgery (n = 186). CTCs were tested before and after surgery as well as during the follow-up period afterwards. In total 495 CTC testing in duplicates were provided. To enrich CTCs, a size-based separation protocol and tube MetaCell® was used. CTCs presence was evaluated by single cell cytomorphology based on vital fluorescence microscopy. Additionally, to standardly applied fluorescence stains, CTCs viability was controlled by mitochondrial activity. CTCs were detected independently on the sampling order in up to 86.7% of the tested blood samples in patients undergoing RCC surgery. There is higher probability of CTC detection with growing tumor size, especially in clear cell renal cell cancer (ccRCC) cases. Similarly, the tumor size corresponds with metastasis presence and lymph node positivity and CTC detection. This paper describes for the first-time successful analysis of viable CTCs and their mitochondria as a part of the functional characterization of CTCs in RCC.

• Klíčová slova
• CTCs, MetaCell, PDL-1, culturing, gene expression, immunotherapy, in vitro, renal cancer,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Liver resection is a surgical procedure associated with a high risk of hepatic failure that can be fatal. One of the key mechanisms involves ischemia-reperfusion damage. Building on the well-known positive effects of hydrogen at mitigating this damage, the goal of this work was to demonstrate the antioxidant, anti-inflammatory, and anti-apoptotic effects of inhaled hydrogen in domestic pigs during major liver resection. METHODS: The study used a total of 12 domestic pigs, 6 animals underwent resection with inhaled hydrogen during general anesthesia, and 6 animals underwent the same procedure using conventional, unsupplemented, general anesthesia. Intraoperative preparation of the left branch of the hepatic portal vein and the left hepatic artery was performed, and a tourniquet was applied. Warm ischemia was induced for 120 minutes and then followed by liver reperfusion for another 120 minutes. Samples from the ischemic and non-ischemic halves of the liver were then removed for histological and biochemical examinations. RESULTS: An evaluation of histological changes was based on a numerical expression of damage based on the Suzuki score. Liver samples in the group with inhaled hydrogen showed a statistically significant reduction in histological changes compared to the control group. Biochemical test scores showed no statistically significant difference in hepatic transaminases, alkaline phosphatase (ALP), lactate dehydrogenase (LD), and lactate. However, a surprising result was a statistically significant difference in gamma-glutamyl-transferase (GMT). Marker levels of oxidative damage varied noticeably in plasma samples. CONCLUSIONS: In this experimental study, we showed that inhaled hydrogen during major liver resection unquestionably reduced the level of oxidative stress associated with ischemia-reperfusion damage. We confirmed this phenomenon both histologically and by direct measurement of oxidative stress in the organism.

• Klíčová slova
• Ischemia-reperfusion injury, hepatic impairment, oxidative damage markers, oxidative stress, reactive oxygen species (ROS),
• Publikační typ
• časopisecké články MeSH

Circulating tumor cells (CTCs), detached from the primary tumor or metastases and shed in the patient's bloodstream, represent a relatively easily obtainable sample of the cancer tissue that can indicate the actual state of cancer, and their evaluation can be repeated many times during the course of treatment. As part of liquid biopsy, evaluation of CTCs provides a lot of clinically relevant information, which reflects the actual, real-time conditions of the disease. CTCs can be used in cancer diagnosis or screening, real-time long-term disease monitoring and even therapy guidance. Their analysis can include their number, morphology, and biological features by using immunocytochemistry and all "-omic" technologies. This review describes methods of CTC isolation and potential clinical utilization in lung cancer.

• Klíčová slova
• Circulating tumor cells, biomarker, culturing, liquid biopsy, lung cancer, review,
• MeSH
• časná detekce nádoru MeSH
• diagnostické techniky molekulární MeSH
• lidé MeSH
• nádorové biomarkery MeSH
• nádorové cirkulující buňky metabolismus   patologie   MeSH
• nádory plic diagnóza   etiologie   terapie   MeSH
• tekutá biopsie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• nádorové biomarkery MeSH