Feeding-Regime-Dependent Intestinal Response of Rainbow Trout after Administration of a Novel Probiotic Feed.

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-ß, 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.

Nephrocalcinosis in farmed salmonids: diagnostic challenges associated with low performance and sporadic mortality.

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.

Trophic distribution of mercury from an abandoned cinnabar mine within the Záskalská reservoir ecosystem (Czech Republic).

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.

Foodborne fluoxetine impacts the immune response in rainbow trout (Oncorhynchus mykkis).

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.

Analysis of Cumulative Fluid Balance Impact on the Stability of Gastrointestinal Tract Anastomosis.

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.

Gene Expression of Antioxidant Enzymes in the Resected Intestine in Crohn's Disease.

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.

A porcine model of endothelial glycocalyx damage by enzymatic digestion: A pilot study.

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.

Detection of Circulating Tumor Cells in Renal Cell Carcinoma: Disease Stage Correlation and Molecular Characterization.

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.

Circulating Tumor Cells in Diagnosis and Treatment of Lung Cancer.

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.

Inhalation of molecular hydrogen prevents ischemia-reperfusion liver damage during major liver resection.

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.

Appendicitis as a rare cause of mechanical small-bowel obstruction: A literature review of case reports.

INTRODUCTION: Although bowel paralysis accompanying acute appendicitis is well known, mechanical bowel obstruction as a direct consequence of appendicitis remains a rare, but potentially life-threatening, acute abdomen. The aim of our literature review was to find all documented cases of this particular complication and compare them with our own case study. METHODS: We searched the PubMed database for relevant articles published from 1963 to 2015. The study included patients for whom direct links between appendicitis and strangulation of the terminal ileum were found, and for which the disease course had been documented in detail. The study also included our own case report since it met the inclusion criteria. A total of 190 articles were examined with a final yield of 17 case reports from 13 articles. RESULTS: 17 patients (11 men and 6 women), with a mean age of 48±23.9years, met the inclusion criteria. The average period between symptom onset and surgery was 3.4±3.7 days. Symptoms of the disease were consistent with small-bowel obstruction. Treatment included simple appendectomy (n=7), possibly supplemented by segmental resection (n=5), followed by ileocecal resection (n=4), and one case that required a right-sided hemicolectomy (n=1). CONCLUSION: We found mechanical bowel obstruction directly related to appendiceal inflammation to be extremely rare, and relatively few individual cases involving this potentially life-threatening complication have been documented in the literature. Clinical signs of the disease are variable, non-uniform, and consistent with symptoms of small-bowel obstruction during their progression.

CT/MRI pelvimetry as a useful tool when selecting patients with rectal cancer for transanal total mesorectal excision.

BACKGROUND: The quality of a total mesorectal excision (TME) is one of the most important prognostic factors for local recurrence of rectal carcinoma. The aim of this study was to clarify the manner in which lesser pelvis dimensions affect the quality of TME via the transabdominal approach, while simultaneously defining the criteria for selecting patients most likely to have Grade 3 TME outcomes for a transanal approach using the TaTME technique. METHODS: An analysis from the registry was conducted using 93 of total 198 patients with rectal cancer of the mid- and lower third of the rectum who underwent: (1) a low anterior resection, (2) an ultra-low resection with coloanal anastomosis, or (3) an intersphincteric rectal resection, all with total mesorectal excision. The procedures were carried out at the Department of Surgery at the University Hospital Hradec Králové between 2011 and 2014. Rectal specimens were histopathologically examined according to a standardized protocol. Pelvimetry data were obtained using anteroposterior, transverse, and sagittal CT or MRI scans. RESULTS: A correlation was found between the quality of the TME and pelvimetry parameter A5, i.e., the angle between the longitudinal axis of the symphysis, and the lines between the symphysis and the promontory (R(2) = -0.327, p < 0.001). The ordinal regression method was used to identify parameters of the model describing levels of probability for TME quality. These relationships were described by equations that provide probability of the achievement of each grade of TME. CONCLUSION: The correlation described by obtained equations between pelvimetry parameters and the quality of TME represents a new tool for use in preoperative decision-making with regard to resection via the transanal approach (TaTME).