Publications by authors named "Stefan Toth"

28 Publications

  • Page 1 of 1

Comparison in detection of prodromal Parkinson's disease patients using original and updated MDS research criteria in two independent cohorts.

Parkinsonism Relat Disord 2021 Jun 30;87:48-55. Epub 2021 Apr 30.

Department of Neurology, P. J. Safarik University, Trieda SNP 1, 04011, Kosice, Slovak Republic; Department of Neurology, University Hospital L. Pasteur, Rastislavova 43, 04190, Kosice, Slovak Republic.

Introduction: MDS research criteria for prodromal Parkinson's disease (pPD) were published in 2015 and updated in 2019. We aimed to determine the difference in pPD patient detection rates in two cohorts recruited via gastrointestinal symptoms (PARCAS study) and the presence of a probable REM sleep behaviour disorder (PDBIOM study) using the original and updated criteria.

Methods: We evaluated all risk and prodromal markers, except genetic testing, plasma urate and physical inactivity, in both cohorts and DaT scan, diabetes mellitus type II and cognitive deficit in the PARCAS cohort. Thresholds of 50% probability for possible pPD and 80% for probable pPD were used.

Results: PPD status as identified by the original/updated criteria showed differences for probable pPD (n = 8/9; original/updated criteria) and possible pPD (n = 9/13) in the PARCAS cohort (total n = 158), as well as for probable pPD (n = 19/21) and possible pPD (n = 6/3) in the PDBIOM cohort (total n = 48). A high concordance rate was found between the two criteria sets (p < 0.001 for all groups).

Conclusion: All probable pPD cases remained in the same category after evaluation with both criteria; three possible pPD cases based on the original criteria exceeded the threshold for probable pPD based on the updated criteria, and five possible new pPD cases were detected, with only one shift in the opposite direction. The updated MDS pPD research criteria tend to identify more patients as positive, yet their accuracy needs to be determined in prospective studies.
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http://dx.doi.org/10.1016/j.parkreldis.2021.04.028DOI Listing
June 2021

Potential influence of prenatal 2.45 GHz radiofrequency electromagnetic field exposure on Wistar albino rat testis.

Histol Histopathol 2021 Mar 25:18331. Epub 2021 Mar 25.

Institute of Neurobiology of Biomedical Research Center of Slovak Academy of Sciences, Kosice, the Slovak Republic.

An ever-increasing use of wireless devices over the last decades has forced scientists to clarify their impact on living systems. Since prenatal development is highly sensitive to numerous noxious agents, including radiation, we focused on the assessment of potential adverse effects of microwave radiation (MR) on testicular development. Pregnant Wistar albino rats (3 months old, weighing 282±8 g) were exposed to pulsed MR at a frequency of 2.45 GHz, mean power density of 2.8 mW/cm², and a specific absorption rate of 1.82 W/kg for 2 hours/day throughout pregnancy. Male offspring were no longer exposed to MR following birth. Samples of biological material were collected after reaching adulthood (75 days). In utero MR exposure caused degenerative changes in the testicular parenchyma of adult rats. The shape of the seminiferous tubules was irregular, germ cells were degenerated and often desquamated. The diameters of the seminiferous tubules and the height of the germinal epithelium were significantly decreased (both at ∗∗p<0.01), while the interstitial space was significantly increased (∗∗p<0.01) when compared to the controls. In the group of rats prenatally exposed to MR, the somatic and germ cells were rich in vacuoles and their organelles were often altered. Necrotizing cells were more frequent and empty spaces between Sertoli cells and germ cells were observed. The Leydig cells contained more lipid droplets. An increased Fluoro Jade - C and superoxide dismutase 2 positivity was detected in the rats exposed to MR. Our results confirmed adverse effects of MR on testicular development.
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http://dx.doi.org/10.14670/HH-18-331DOI Listing
March 2021

Vaccines Targeting PSCK9 for the Treatment of Hyperlipidemia.

Cardiol Ther 2020 Dec 31;9(2):323-332. Epub 2020 Jul 31.

MEDIPARK, University Research Park for Preclinical and Clinical Research, Pavol Jozef Safarik University, Trieda SNP 1, 040 11, Kosice, Slovakia.

Despite progress in both primary and secondary prevention, cardiovascular diseases (CVD) are still the largest group of ailments contributing to morbidity and mortality worldwide. Atherosclerotic changes, the primary pathological substrate for CVD, are closely related to hypercholesterolemia. Therefore, the treatment of hypercholesterolemia is a key therapeutic strategy for CVD management. Statins, as the gold standard in the treatment of hypercholesterolemia, have shown enhanced cardiac outcomes in many randomized clinical trials. However, often despite the maximum allowed and tolerated dosage of statins, we are not able to reach the target cholesterol levels, and thus patients persist at an increased cardiovascular risk. Recently, most of the large clinical studies in the field of preventive cardiology have focused on proprotein convertase subtilisin kexin type 9 (PCSK9) and its activity regulation. PCSK9 plays an essential role in the metabolism of LDL particles by inhibiting LDL receptor recirculation to the cell surface. Recent studies have shown that inhibition of PCSK9 by the administration of monoclonal antibodies is capable of significantly reducing LDL levels (up to an additional 60%) as well as reducing the incidence of CVD. However, this treatment procedure of administering the anti-PCSK9 antibodies, most frequently two times a month, has its limitations in terms of time, patient adherence, and nevertheless cost. Administration of active vaccination instead of passive immunization with anti-PCSK9 antibodies may be an effective way of controlling blood cholesterol levels. However, clinical data, as well as human testing, are still inadequate. This work aims to provide an overview of PCSK9 vaccines and their potential clinical benefit.
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http://dx.doi.org/10.1007/s40119-020-00191-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7394273PMC
December 2020

The effect of Betanin parenteral pretreatment on Jejunal and pulmonary tissue histological architecture and inflammatory response after Jejunal ischemia-reperfusion injury.

Exp Mol Pathol 2019 10 1;110:104292. Epub 2019 Aug 1.

Department of Internal Medicine, Borthers of Mercy Hospital, Brno, Czech Republic; 2nd Department of Surgery, Faculty of Medicine, Masaryk University and St. Anne's University Hospital, Brno, Czech Republic. Electronic address:

Intestinal ischemic-reperfusion (IR) injury has detrimental effects on both local and distant organs in the body. Betanin is known for its antioxidant properties, and it is found mostly in vegetables. Therefore, the aim of the present study was to test the hypothesis that betanin administration prior intestinal IR, may be beneficial in protecting jejunal mucosa and lung parenchyma against IR damage. Male specific pathogen-free Charles River Wistar rats were used (n = 42). Betanin (50 mg/kg) was administered intraperitoneally 30 min before ischemia of the superior mesenteric artery lasting 1 h, followed by 1, 4 and 24 h of reperfusion. Immunohistochemical as well as histomorphometrical analysis indicated a protective effect of betanin pretreatment on jejunal tissue. Regarding morphometrical analysis betanin significantly (p < 0.01) augments intestinal villus height after 24 of reperfusion comparing to early stages. Betanin application reduced number of mast cells population in early reperfusion periods (p < 0.05). The protective effect of betanin on lung parenchyma, was detected in late reperfusion period (24 h) with improvement of histopathological injury index and morphometric analysis (p < 0.001 for both). The improvement of histopathological injury index (p < 0.001) and morphometric analysis (p < 0.001) during the late reperfusion period, suggests a protective effect of betanin on lung parenchyma. Moreover, suppression of the inflammatory response was mirrored by the reduction of myeloperoxidase (MPO) positive cells within lung parenchyma after 1 and 4 h of reperfusion (p < 0.001). Especially, during the first 4 h of reperfusion after betanin administration, a reduction of 74% of the polymorphonuclear neutrophils infiltration (MPO positive cell population) and of a nearly 46% of active MCs was observed. Upon morphometric examination, the lung histological architecture after 24 h of reperfusion appeared to be almost 100% better following betanin treatment, with 25% thinner interalveolar septa and 20% larger alveolar surface for respiratory gas exchange. The results suggest that betanin pretreatment protects the jejunal mucosa and the lung parenchyma, as well as reduces the inflammatory cell density after intestinal IR injury.
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http://dx.doi.org/10.1016/j.yexmp.2019.104292DOI Listing
October 2019

The effect of 2.45 GHz non-ionizing radiation on the structure and ultrastructure of the testis in juvenile rats.

Histol Histopathol 2019 Apr 27;34(4):391-403. Epub 2018 Sep 27.

Department of Anatomy, Histology and Physiology, Institute of Histology and Embryology, University of Veterinary Medicine and Pharmacy in Košice, Slovakia.

Background: Nowadays, mobile devices that emit non-ionizing electromagnetic radiation (EMR) are predominantly used by juveniles and pubescents. The aim of the present study was to evaluate the effect of whole body pulsed EMR on the juvenile Wistar albino rat testis at a frequency of 2.45 GHz and mean power density of 2.8 mW/cm².

Methods: The investigated animals (n=24) were divided into two control and two EMR groups (5 and 6 week old rats; 6 rats per group). Both EMR groups were irradiated continually for 3 weeks (2h/day) from postnatal days 14 and 21, respectively.

Results: EMR caused an irregular shape of seminiferous tubules with desquamated immature germ cells in the lumen, a large number of empty spaces along the seminiferous epithelium and dilated and congested blood vessels in the interstitial tissue of the testis. The cytoplasm of Sertoli cells showed strong vacuolization and damaged organelles, with the cytoplasm full of different heterophagic and lipid vacuoles or the cytoplasm of spermatocytes with swollen mitochondria in both irradiated groups. A significant increase in the total tubular area of seminiferous tubules was observed in both EMR groups compared with controls (P<0.001). A significant increase in the TUNEL-positive apoptotic nuclei (P<0.01) was accompanied by a significant rise in both Cu-Zn-SOD (P<0.01) and Mn-SOD (P<0.001) positive cells in the 6 week old experimental rats compared to control animals.

Conclusion: Our results confirmed a harmful effect of non-ionizing radiation on the structure and ultrastructure of the juvenile rat testis.
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http://dx.doi.org/10.14670/HH-18-049DOI Listing
April 2019

Detection of anti-infliximab antibodies in Slovak IBD patients and its costs saving effect.

Neuro Endocrinol Lett 2017 Nov;38(Suppl1):5-9

1st Department of Internal Medicine, and Louis Pasteur University Hospital, Slovakia.

Objective: Management chronic inflammatory bowel disease (IBD) patients is associated with diagnosis, targeted treatment and and individual approach. There is a group of patients which loss the response to the biologic treatment caused by insufficient levels of biologics or positive antibodies against these drugs. This study was aimed to determine the prevalence of patients with positive antibodies against the biological treatment and the costs saving probabilities of the antibodies detection during the treatment.

Study Design: This retrospective study was based on examination of 183 IBD patients' sera (72 with Crohn's disease (CD) and 111 ulcerative colitis (UC)) treated with infiliximab.

Methods: Circulating serum infliximab concentrations and anti-infliximab antibodies (ATI) were quantified by ELISA methods. Costs associated with the treatment were analysed from the data of General Health Insurance Company, Slovakia.

Results: The average infliximab concentrations in groups of CD were 2.9 µg/mL, 38.9% of samples had a concentration ≤1 µg/mL. Group with UC had average infliximab levels of 3.19 µg/mL, 32.4% bellow ≤1 µg/mL. Positive ATI levels were detected in 52 patients, in 28 patients with CD (38.8%) and 24 patients with UC (21.6%). The average values of the antibodies were 387.75 U/ml in CD and 391.94 U/ml in UC group. More than 28% IBD patients were positive for ATI. After application of the results to the database of all IBD patients, finishing of the treatment with ATI could lead (after considering the ATI quantification costs) to possible annual savings of more than €2 million in Slovakian health-care system.

Conclusions: Monitoring of infliximab and antibodies against infliximab and anti-TNF-α biologics may help optimize treatment strategies and costs for biological treatment.
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November 2017

Quercetin protects jejunal mucosa from experimental intestinal ischemia reperfusion injury by activation of CD68 positive cells.

Acta Histochem 2018 Jan 10;120(1):28-32. Epub 2017 Nov 10.

Department of Chemical Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic; 2nd Department of Surgery, Faculty of Medicine, Masaryk University and St. Anne's University Hospital, Brno, Czech Republic. Electronic address:

The aim of our study was to analyse the possible protective effect of quercetin application during the jejunal ischemia-reperfusion injury (IRI) in rats. Quercetin was administered intraperitoneally 30min before 1h ischemia of superior mesenteric artery with following 24h lasting reperfusion period. The male specific pathogen-free (SPF) Charles River Wistar rats were used. In the group with applied quercetin, the significantly increased (p<0.001) levels of anti-inflammatory cytokine IL10 were observed both in the blood serum and jejunal tissue. The improvement of the mucosal tissue morphology and proliferating and DNA repairing cell number measured by PCNA activity were recorded by more than 30% higher in the quercetin group. Simultaneously, significant elongation of the intestinal glands (p<0.001) and increase in the number of CD68-positive cells in the lamina propria mucosae (p<0.001) in comparison with control group were found. Based on our results, the preventive application of quercetin before induction of jejunal IRI stimulates faster jejunal mucosa restoration and it seems to have immunomodulatory and anti-inflammatory effects as well. CD68-positive macrophages could have crucial role in this process since they work as both growth factor and cytokine producers.
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http://dx.doi.org/10.1016/j.acthis.2017.11.001DOI Listing
January 2018

Quercetin attenuates the ischemia reperfusion induced COX-2 and MPO expression in the small intestine mucosa.

Biomed Pharmacother 2017 Nov 12;95:346-354. Epub 2017 Sep 12.

Department of Chemical Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic. Electronic address:

Quercetin, the active substance of tea, fruits and vegetables, exerts a broad spectrum of pharmacological activities and is considered to have potential therapeutic application. The present study was designed to investigate the beneficial effect of quercetin against experimental ischemia- reperfusion (IR) injury of the small intestine in rats. Quercetin was administrated intraperitoneally 30min before 1h ischemia of superior mesenteric artery with following reperfusion periods lasting 1, 4 and 24h. The male specific pathogen-free Charles River Wistar rats were used (n=45). In acute phase, 4h after start of reperfusion, the quercetin induced a significant decrease in mucosal injury index (p<0.05) accompanied by a significant decrease in cyclooxygenase-2 (COX-2) expression in the epithelial lining of the intestinal villi in comparison with the control group (p<0.01). In the epithelium of the intestinal glands, COX-2 expression resulting from IR injury significantly increased regardless quercetin application (in control group p<0.001; in quercetin group p<0.05), but in quercetin group, significant decrease in it during 24h of reperfusion in a late phase of IR injury was detected (p<0.001). Based on morphology of COX-2 positive cells, the COX-2 positivity was found particularly in goblet cells of the intestinal villi epithelium and enteroendocrine cells respectively, in the glandular epithelium. We concluded that quercetin application attenuated mucosal damage from IR injury by inhibiting neutrophil infiltration which was demonstrated by a lower number of myeloperoxidase positive cells in the lamina propria during both phases of IR injury and the significant decrease in that in a late phase after 24h of reperfusion (p<0.05).
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http://dx.doi.org/10.1016/j.biopha.2017.08.038DOI Listing
November 2017

Elevated Circulating PCSK9 Concentrations Predict Subclinical Atherosclerotic Changes in Low Risk Obese and Non-Obese Patients.

Cardiol Ther 2017 Dec 16;6(2):281-289. Epub 2017 Jun 16.

1st Department of Internal Medicine, Faculty of Medicine, Pavol Jozef Safarik University, Trieda SNP 1, 041 90, Košice, Slovakia.

Introduction: Many studies have highlighted the important role of PCSK9 in the development of cardiometabolic changes and its possible function as a biomarker of myocardial infarction or ischemic heart disease. This study aimed to determine the relationship between circulating PCSK9 levels and subclinical vascular changes in the group of low risk patients without manifest cardiovascular diseases.

Methods: In this study, 120 healthy patients, free of manifest cardiovascular diseases, diabetes mellitus, and without lipid-lowering therapy, were divided into three groups based on BMI: normal weight (N = 50), overweight (N = 30), and obese (N = 40). Biochemical parameters, including basic lipid and non-lipid ones, were analyzed. PCSK9 levels were measured by ELISA, vascular changes were quantified by carotid ultrasound (carotid artery intima-media thickness, cIMT), and arterial stiffness parameters (pulse wave velocity, PWV; augmentation index, AI; stiffness parameter, β) were measured by an echo-tracking method.

Results: Plasma levels of PCSK9 significantly increased in obese (172.78 ± 51.67 ng/mL) in comparison with overweight (120.14 ± 37.64, p < 0.001) and normal weight groups (114.92 ± 35.87, p < 0.001). Differences between the overweight and normal weight groups were not significant (p = 0.85). The level of PCSK9 significantly correlated with values of BMI (p < 0.001, r = 0.38). In addition to increase in laboratory parameters associated with moderate metabolic changes, significant increase in cIMT and parameters of vascular changes (β, AI, PWV) were detected in groups with elevated BMI. Significant positive linear correlation of PCSK9 concentrations and cIMT (p < 0.001, r = 0.39), PWV (p < 0.001, r = 0.31), and β (p < 0.001, r = 0.3) were found. In multivariable regression analysis after adjusting for gender, age, BMI, and LDL, the impact of PCSK9 on cIMT, β, and PWV remained significant (p = 0.006, 0.03, and 0.002, respectively).

Conclusion: PCSK9 plasma levels significantly correlated with subclinical vascular changes and their values were significantly elevated in obese subjects. We assume that PCSK9 could be used as a predictor of early vascular involvement, prior to the existence of manifest atherosclerosis. These results also highlight the role of anti-PCSK9 treatment in primary prevention.
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http://dx.doi.org/10.1007/s40119-017-0092-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688969PMC
December 2017

Addition of omega-3 fatty acid and coenzyme Q10 to statin therapy in patients with combined dyslipidemia.

J Basic Clin Physiol Pharmacol 2017 Jul;28(4):327-336

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Background: Statins represent a group of drugs that are currently indicated in the primary and secondary prevention of cardiovascular events. Their administration can be associated with side effects and the insufficient reduction of triacylglyceride (TAG) levels. This study aimed to assess the effect of the triple combination of statins with omega-3 fatty acids and coenzyme Q10 (CoQ10) on parameters associated with atherogenesis and statin side effects.

Methods: In this pilot randomized double-blind trial, 105 subjects who met the criteria of combined dislipidemia and elevated TAG levels were randomly divided into three groups. In the control group, unaltered statin therapy was indicated. In the second and third groups, omega-3 PUFA 2.52 g/day (Zennix fa Pleuran) and omega-3 PUFA 2.52 g+CoQ10 200 mg/day (Pharma Nord ApS) were added, res//. At the end of the 3-month period (±1 week), all patients were evaluated.

Results: Significant reduction of hepatic enzymes activity, systolic blood preasure, inflammatory markers and TAG levels were detected in both groups in comparison to the control group. Activity of SOD and GPx increased significantly after additive therapy. Coenzyme Q10 addition significantly reduced most of the abovementioned parameters (systolic blood preasure, total cholesterol, LDL, hsCRP, IL-6, SOD) in comparison with the statin+omega-3 PUFA group. The intensity of statin adverse effects were significantly reduced in the group with the addition of CoQ10.

Conclusions: The results of this pilot study suggest the possible beneficial effects of triple combination on the lipid and non-lipid parameters related to atherogenesis and side effects of statin treatment.
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http://dx.doi.org/10.1515/jbcpp-2016-0149DOI Listing
July 2017

Rare Presentation of Left Lower Lobe Pulmonary Artery Dissection.

Case Rep Med 2017 5;2017:2760535. Epub 2017 Jan 5.

First Department of Internal Medicine, Louis Pasteur University Hospital, Trieda SNP 1, 041 90 Košice, Slovakia.

. Pulmonary arterial dissection with chronic pulmonary arterial hypertension as its major cause is a very rare but life-threatening condition. In most cases the main pulmonary trunk is the affected site usually without involvement of its branches. Segmental or lobar pulmonary artery dissection is extremely rare. . We report a unique case of left lower lobe pulmonary artery dissection in a 70-year-old male, with confirmed chronic pulmonary hypertension. To confirm dissection MDCT pulmonary angiography was used. Multiplanar reformation (MPR) images in sagittal, coronal, oblique sagittal, and curved projections were generated. This case report presents morphologic CT features of rare chronic left lobar pulmonary artery dissection associated with chronic pulmonary hypertension at a place of localised pulmonary artery calcification. CT pulmonary angiography excluded signs of thromboembolism and potential motion or flow artefacts. . To the best of our knowledge, no case of lower lobe pulmonary artery dissection with flap calcification has been reported yet. CT imaging of the chest is a key diagnostic tool that is able to detect an intimal flap and a false lumen within the pulmonary arterial tree and is preferred in differential diagnosis of rare complications of sustained pulmonary arterial hypertension.
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http://dx.doi.org/10.1155/2017/2760535DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5244010PMC
January 2017

Protective effect of ischemic preconditioning on the jejunal graft mucosa injury during cold preservation.

Exp Mol Pathol 2015 Oct 27;99(2):229-35. Epub 2015 Jun 27.

International Clinical Research Center, St. Anne's University Hospital and Masaryk University, Brno, Czech Republic. Electronic address:

Protection of intestinal graft mucosa during cold preservation is still an unmet need in clinical practice, thus affecting the success of transplantation. The present study investigates the ability of two ischemic preconditioning (IPC) procedures to limit cold preservation injury. Three groups of Sprague-Dawley rats were recruited (n=11 each) as follows: the short IPC (SIPC) performed through 4 cycles of mesenteric ischemia of 4 min each followed by 10 min of reperfusion, the long IPC (LIPC) obtained by 2 ischemic cycles of 12 min each followed by 10 min of reperfusion, and the control group (C) without IPC. Grafts were then stored in cold histidine-tryptophan-ketoglutarate solution and samples were taken at 0, 3, 6 and 9 h lasting preservation. Both IPC groups showed an advanced degree of preservation with delayed development of graft mucosa damage, mainly in the crypt region. At the beginning of preservation, the graft mucosa in both IPC groups showed lower degree of mucosal injury index (MII) by 50% in comparison with C group. Specifically, a significant improvement of MII was observed after 3h of preservation in the LIPC group (p<0.05) in comparison with untreated C grafts. Significant atrophy of the intestinal mucosa in C group was found after 3h of preservation (p<0.01), in SIPC group the progress of atrophy was delayed to 6 h (p<0.001), and in LIPC group only moderate decrease in that was found. A parallel increase of laminin expression with the MII rate after 6 and 9h of preservation in comparison with the level at time 0 was observed in all grafts (p<0.001 and p<0.01, respectively). In both IPC groups the apoptotic cell (AC) rate was significantly reduced at the beginning of cold preservation (p<0.05 both). Moreover, in both the SIPC and C groups, the progressive increase in MII rate connected with AC rate decrease was due to a predominance of necrosis. By contrast in the LIPC group, after an increase of nearly 50% in the AC rate at the 3rd hour, its level remained fairly constant during the further 6 h of preservation, thus probably preventing necrosis and improving graft viability.
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http://dx.doi.org/10.1016/j.yexmp.2015.06.020DOI Listing
October 2015

Influence of dietary supplementation with flaxseed and lactobacilli on the mucosal morphology and proliferative cell rate in the jejunal mucosa of piglets after weaning.

Int J Exp Pathol 2015 Jun 1;96(3):163-71. Epub 2015 May 1.

Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy, Kosice, Slovak Republic.

The aim of the study was to investigate the influence of flaxseed and lactobacilli supplementation to the diet of piglets during the time period between 10 days before and 21 days after weaning. The morphometry of the jejunal mucosa and proliferative ratio of both epithelial and lamina propria cells were compared with those found in a group of piglets fed with the usual diet added with sunflower oil during the same time period. The addition of flaxseed oil to the diet significantly increased the crypt depth in comparison with both groups supplemented with sunflower (P < 0.05 and 0.001 respectively) on the weaning day. Moreover, the flaxseed addition caused a significant decrease in villus height (P < 0.01) and crypt depth (P < 0.01) 21 days postweaning in comparison with the sunflower group. The proliferative ratio of the epithelial cells in the sunflower group on the weaning day was significantly higher than in both flaxseed groups (P < 0.01). Paradoxically, significantly higher proliferative activity in the mucosal connective tissue in the group with flaxseed supplementation in comparison with the sunflower group was observed on the day of weaning, as well as 3 days later (P < 0.05 both). A combination of flaxseed with lactobacilli showed significantly lower proliferative activity in the connective tissue cells from weaning up to 7 days after weaning (P < 0.05 all) in comparison with the flaxseed group.
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http://dx.doi.org/10.1111/iep.12129DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4545427PMC
June 2015

Influence of dietary supplementation with flaxseed and lactobacilli on the cells of local innate immunity response in the jejunal mucosa in piglets after weaning.

Acta Histochem 2015 Mar 9;117(2):188-95. Epub 2015 Jan 9.

Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy, Kosice, Slovak Republic.

A histological study was designed to determine the influence of flaxseed and/or lactobacilli inclusion in the diet of piglets from 10 days before to 21 days after weaning. The selected inflammatory cell population incidence in the piglet jejunal mucosa was investigated. Significantly higher numbers of myeloperoxidase-positive (P<0.01) and CD163-positive (P<0.001) cells in the jejunal mucosa were recorded on the weaning day and for 7 days after (P<0.001 and P<0.01, respectively) in the flaxseed group compared with the basal diet. The number of intraepithelial lymphocytes was also significantly increased until 3 days after weaning (P<0.001). A prolonged significant increase in the myeloperoxidase-positive cells and intraepithelial lymphocyte numbers in the flaxseed+lactobacilli group was detected. In contrast, the number of CD163-positive cells in the flaxseed+lactobacilli group was significantly lower on the day of weaning (P<0.05) and 3 days after (P<0.01). The same effect was observed in the group with lactobacilli alone during the first 3 days after weaning (P<0.05 and P<0.01, respectively) and these findings indicate down-regulation of CD163 expression in the jejunal mucosa by lactobacilli. The presence of lactobacilli in the diet had a stimulatory effect on goblet cell quantity in the epithelium (P<0.001) and a distinct 50% reduction in the flaxseed group (P<0.01) compared with the basal diet was observed on the weaning day. A significant increase in myeloperoxidase-positive cell number in the jejunal mucosa in the flaxseed+lactobacilli group was the only significant difference (P<0.05 and P<0.01, respectively) found 21 days after weaning in comparison with all the other groups, indicating the pro-inflammatory effect of this feed additive combination. We conclude that dietary supplementation with flaxseed and lactobacilli on the cells of local innate immunity response in the jejunal mucosa in piglets after weaning might be linked with significant anti-inflammatory effects in the jejunal mucosa.
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http://dx.doi.org/10.1016/j.acthis.2014.12.005DOI Listing
March 2015

Impact of alanyl-glutamine dipeptide on proliferative and inflammatory changes in jejunal mucosa after acute mesenteric ischemia.

J Pediatr Surg 2014 Sep 10;49(9):1385-9. Epub 2014 Feb 10.

Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University, Šrobárova 2, Košice, Slovak Republic.

Purpose: The aim of our study was to determinate the impact of dipeptide (alanyl-glutamine) administration on inflammatory and proliferative changes in jejunal mucosa after acute mesenteric ischemia.

Methods: Male Wistar rats (n=30) were divided into three groups: ischemia/reperfusion (IR) group which undergoes 60min of mesenteric ischemia and 1 or 24h of reperfusion (IR1, IR24, n=12). Groups with dipeptide administration (D+IR1, D+IR24, Dipeptiven con inf., i.v., 0.75 g/kg) prior to IR injury were followed by 1 and 24h of reperfusion. At the end of reperfusion period jejunal bioptic samples were obtained for histological (H&E), histochemical (Alcian blue) and immunohistochemical (anti-PCNA, anti-MPO) evaluations.

Results: Our results pointed out a significant (p<0.001) increase of histopathological injury score in IR1 group compared to D+IR1 group. Immunohistochemical evaluation showed that MPO-positivity was significantly increased in IR groups after 1 (p<0.001) as well as 24h of reperfusion (p<0.01) compared to dipeptide pretreated groups. Proliferative/reparatory rate was assessed using anti-PCNA antibody and showed a significant increase (p<0.01) in PCNA cell positivity in lamina propria in dipeptide treated group compared to IR group.

Conclusion: In conclusion we may suggest that administration of alanyl-glutamine dipeptide prior to IR injury may help to protect small intestine and its mucous membrane integrity against insult such as intestinal ischemic/reperfusion injury presents.
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http://dx.doi.org/10.1016/j.jpedsurg.2014.01.056DOI Listing
September 2014

Immunohistochemical expression of MPO, CD163 and VEGF in inflammatory cells in acute respiratory distress syndrome: a case report.

Int J Clin Exp Pathol 2014 15;7(7):4539-44. Epub 2014 Jun 15.

Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University Šrobárova 2, Košice, Slovak Republic.

Acute respiratory distress syndrome (ARDS) is a serious medical condition occurring in patients with polytrauma, pulmonary or non-pulmonary sepsis, pneumonia and many other circumstances. It causes inflammation of the lung parenchyma leading to impaired gas exchange with a systemic release of inflammatory mediators, causing consequential lung tissue injury, hypoxemia and frequently multiple organ failure. The aim of current study was to describe expression of inflammatory markers (myeloperoxidase, CD163 and vascular endothelial growth factor) by the cells in acute phase of ARDS. The lung samples of a 20-year-old man who had suffered a serious motorbike accident were obtained for histological examination. He died on the seventh day as a consequence of respiratory failure. Our results imply that expression of CD163 was restricted to activated alveolar macrophages and monocytes. Immunopositivityof MPO was observed in neutrophil granulocytes within lung alveoli and lung blood vessels. Myeloperoxidase positivity was observed in alveolar macrophages, too. Vascular endothelial growth factor was expressed in cytoplasm of neutrophil granulocytes, monocytes, small-sized alveolar macrophages and type II pneumocytes localized mostly inside lung alveoli. On the contrary, no positivity was observed in lung endothelial cells of blood vessels.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4129085PMC
May 2015

Trehalase as a possible marker of intestinal ischemia--reperfusion injury.

Acta Biochim Pol 2013 20;60(3):411-6. Epub 2013 Sep 20.

Department of Medical and Clinical Biochemistry and LABMED, and Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University, Košice, Slovakia.

Background: Different pathological affections of the small intestine cause corresponding morphological and functional changes. The present study was aimed to assess intestinal trehalase activities during ischemia and following reperfusion, correlate them with the pathological changes and determine whether trehalase could be used as a biochemical marker of the intestinal ischemia, ischemia - reperfusion injury.

Material And Methods: Wistar rats, randomly divided into 5 experimental groups (IR) (each n=15), were subjected to one hour mesenteric ischemia followed by 0, 1, 4, 12 and 24 hours of reperfusion. As a control group sham operated animals were used (n=15). The activity of trehalase was determined using an adapted Dahlqwist method. The range of intestinal injury was determined using histological (histopathological injury index and goblet cell quantification) and immunohistochemical (Ki67, InSitu TUNEL) methods.

Results: The highest activities of trehalase were recorded in the control group (C=4.42 ± 0.373 μmol/mg/h). The most altered intestinal histology detected in group IR1 was accompanied by the lowest trehalase activity (IR1=0.97 ± 0.209 μmol/mg/h; p < 0.001 C vs. IR1). Improved histological structure in the remaining reperfusion periods correlated with increase in trehalase activity. Almost normal mucosal histological architecture and 72% of the enzymatic activity were restored after 24 hours of reperfusion (IR24=3.20 ± 0.266 μmol/mg/h; p < 0.01 IR1 vs. IR24).

Conclusion: The correlation between intestinal histology and trehalase activities during intestinal injury has been shown. Trehalase activity is closely associated with the status of the histological architecture of the small intestine.
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April 2014

Intravenous administration of tetramethylpyrazine reduces intestinal ischemia-reperfusion injury in rats.

Am J Chin Med 2013 ;41(4):817-29

Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University, Slovakia.

Intestinal ischemia-reperfusion injury (IIRI) is a life-threatening condition requiring prompt medical intervention. Tetramethylpyrazine (TMP) is a biologically active alkaloid isolated from Ligusticum wallichii. Previously, it was shown that TMP causes vasodilatation and inhibition of platelet aggregation as well as exhibits significant antioxidant effects. Therefore, the aim of the present study was to evaluate possible therapeutic effects of TMP in the prevention of IIRI. Wistar rats (n = 80) were randomly divided into eight experimental groups and subjected to a 1 h occlusion of cranial mesenteric artery followed by 0, 1, 12, and 24 h period of reperfusion. Thirty minutes before the IIRI animals received either TMP (30 mg/kg, i.v.) or identical volume of saline. In addition, a control group of 10 animals was not exposed to IIRI. Intestine morphology was evaluated by using histopathological injury index examination (HII), goblet and Paneth cells quantification as well as by applying immunofluorescent methods such as InSitu TUNEL and caspase-3 positivity assessment. Here we showed that preconditioning with TMP prior IIRI decreases the grade of injury. Significant reduction of HII was detected in TMP pretreated groups after 0, 1, and 12 h of reperfusion where injury reduction up to 75% was found. Lower histopathological damage in preconditioned groups was accompanied with increased number of secretory epithelial cells and decreased number of apoptotic cells. These results demonstrate the protective effect of TMP on the small intestine mucosa, suggesting administration of TMP as a molecule for pharmacological intervention against IIRI.
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http://dx.doi.org/10.1142/S0192415X13500559DOI Listing
October 2013

Immunohistochemical study of jejunal graft mucosa cell populations during the initial adaptation phase in the host body in rats.

Acta Histochem 2013 Oct 28;115(8):803-9. Epub 2013 Apr 28.

Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University, Šrobárova 2, 041 80 Košice, Slovakia. Electronic address:

The character of the changes in cell populations within the jejunal graft mucosa during the initial adaptation phase in the host body was investigated. 24 adult male Wistar rats underwent intestinal heterotopic allotransplantation. Aorto-aortal and porto-caval anastomoses were performed using the end-to-side microsurgery technique. Graft tissues were compared to the intestinal tissues of the recipients. This study demonstrates that: (1) Distinct injury to the graft mucosa 1h after transplantation was accompanied by significant reduction in numbers of epithelial secretory cell populations. The injury was more intense in the mesenteric portion. Six hours after transplantation the graft mucosa was covered by a continuous epithelium, but the number of goblet and Paneth cells was found to be less than 30% of that in the recipient epithelium. (2) In comparison with recipients, myeloperoxidase-positive cell numbers increased significantly in the graft mucosa 1 h after transplantation. In the epithelial layer, denudation and destruction of villi was associated with a significant reduction in intraepithelial lymphocyte numbers. A significant decrease in mucosal mast cell numbers was detected 6 h after transplantation. They attained only 10% of the number found in the recipients. (3) Time-dependent changes in the graft mucosa revealed that CD163-positive cells increased significantly in the graft mucosa during 6 h after transplantation and reached the level found in the recipients. In contrast, the myeloperoxidase-positive cell population significantly decreased in the graft mucosa within the initial 6 h.
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http://dx.doi.org/10.1016/j.acthis.2013.03.007DOI Listing
October 2013

The relationship between morphology and disaccharidase activity in ischemia- reperfusion injured intestine.

Acta Biochim Pol 2012 30;59(4):631-8. Epub 2012 Nov 30.

2nd Department of Obstetrics and Gynecology, PJ Šafárik University and L Pasteur University Hospital, Košice, Slovak Republic.

Background: Questions regarding functional markers characterizing injured intestines remain unanswered. Brush border disaccharidases are crucial for the functioning of the intestines.

Aims: The study was designed to assess changes in disaccharidase activity (DA) following intestinal injury and to compare them with morphological changes.

Methods: Wistar rats, randomly divided into six experimental groups (each n = 6), were subjected to different ischemic/reperfusion injury. One-hour mesenteric ischemia followed by reperfusion for 0, 1, 2, 4, 12 or 24 hours was induced. As a control group sham-operated animals were used (n = 6). Intestine morphology was evaluated using histopathological injury index (HII) and goblet cell (GC) detection. DA (sucrase and maltase) was studied in mucosal scrape or in entire intestinal wall samples.

Results: Moderate morphological damage (HII, GC) after mesenteric ischemia was detected. Deepening of the injury was found during reperfusion with a maximum after two hours. Improved morphology with longer reperfusion confirmed reversible damage with almost normal mucosal structure after 24 hours of reperfusion. Similar pattern was observed when DA was measured. The lowest activity was detected after 2 hours of reperfusion followed by increasing activity in the subsequent reperfusion periods. Physiological values after 24 hours of reperfusion were seen only in samples of entire intestinal wall.

Conclusions: Significant changes in intestinal DA were observed after intestinal ischemia/reperfusion injury. A similar pattern was seen for morphological characteristics. Although based on microscopic survey the intestine seems to be fairly regenerated, some functional limitation is expected to persist.
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June 2013

Intestinal ischemia-reperfusion injury - the histopathological status of remote vital organs in acute and subacute phases.

Ann Transplant 2012 Jan-Mar;17(1):11-20

2nd Department of Gynaecology and Obstetrics, Faculty of Medicine, Pavol Jozef Šafárik University, Košice, Slovak Republic.

Background: Improvement of graft recovery and function follows current trends in intestinal transplantation; however, the alteration of remote organs (RO) predicts complicated systemic rejection. This study was conceived to describe the histopathological status of RO arising in both acute and subacute stages after intestinal ischemia-reperfusion injury (IIR) injury.

Material/methods: Wistar rats (n=54) were divided into 7 experimental groups (n=7 each). All the animals were subjected to 60 min mesenteric ischemia and subsequently to reperfusion 2 h, 4 h, 24 h, 72 h, 10 days, 20 days and 30 days following the groups IR2 h, IR4 h, IR24 h, IR72 h, IR10 d, IR20 d and IR30 d. As a control group (S; n=5) sham-operated animals were used. Histopathological scores (HPS) were evaluated in biopsies of the right kidney, heart and colon ascendens.

Results: Statistically significant increase in kidney HPS was seen during reperfusion, with the peak in IR4h group (p<0.01). Thereafter, improved morphology was observed; however, increased HPS was seen even in the subacute stage, and significant deterioration of HPS up to 10 days of reperfusion was detected (p<0.05). Heart biopsies also showed statistically increased HPS value in IR4h group (p<0.05). Intact morphology of the colon was detected in all reperfusion periods.

Conclusions: IIR causes a systemic reaction affecting RO. The peak of alteration for kidney and heart morphology was induced by 60 min of ischemia followed by 4 h of reperfusion. Thereafter, improved morphology was observed, although latent persistence of histopathological changes was seen even in the subacute stage. The colon remained intact during the whole experiment despite its anatomical proximity, confirming its high immunological capacity.
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http://dx.doi.org/10.12659/aot.882631DOI Listing
August 2012

Mesenteric ischemia-reperfusion injury: specific impact on different cell populations within the jejunal wall in rats.

Acta Histochem 2012 May 30;114(3):276-84. Epub 2011 Jun 30.

Department of Histology and Embryology, Pavol Jozef Śafárik University, Košice, Slovakia.

The progress of jejunal damage and recovery in the course of mesenteric ischemia-reperfusion injury in rats at different time periods was investigated. Mesenteric ischemia lasting 1h followed by 1h of reperfusion caused a significant disintegration of the mucosa, reduction of the muscular layer and diminution of the wall thickness. The loss of epithelium included enterocytes, goblet cells and Paneth cells. Paradoxically, increasing numbers of serotonin-producing cells and the beginning of regenerative processes, expressed by significantly higher proliferation, were recorded in the epithelium during this period. Disintegration of connective tissue and massive degranulation of serotonin-positive cells were found in the lamina propria. After 24h of reperfusion, restitution of the mucosa was found, expressed by normal villous morphology and re-epithelialization. However, some parameters were still significantly affected even more than in the acute phase of reperfusion. In the epithelium, decreased numbers of Paneth cells and increased population of serotonin-producing cells were found. The greatest proliferation of connective tissue cells and intensified reduction of the muscular layer were also detected in this reperfusion period. After 30 days of reperfusion, moderate damage remained, but only the increased number of Paneth cells and decreased number of serotonin-producing cells in the lamina propria were significant.
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http://dx.doi.org/10.1016/j.acthis.2011.06.004DOI Listing
May 2012

Different ischemic preconditioning regimens affecting preservation injury of intestines.

Eur Surg Res 2011 28;46(4):207-13. Epub 2011 Apr 28.

Department of Histology and Embryology, Faculty of Medicine, P.J. Šafárik University, Košice, Slovakia.

Decreasing ischemia-reperfusion injury in intestinal transplantation is of paramount importance for improving graft recovery and function. This study explores the ability of two ischemic preconditioning (IPC) regimens to reduce preservation injury. Sprague-Dawley rats were divided into 3 groups (n = 11 each). In the controls (group C), intestinal grafts were harvested and preserved. IPC was performed either through 4 cycles of mesenteric ischemia of 4 min each followed by 10 min of reperfusion (group BIPC) or 2 ischemic cycles of 12 min each followed by 10 min of reperfusion (group LIPC). Grafts were stored in histidine-tryptophan-ketoglutarate, and samples were taken 0, 3, 6, 9, 12, 18, and 24 h after preservation. Preservation injury was scored using the Park/Chiu scale. Goblet cells (GC), enteroendocrine cells (EEC) and serotonin-producing EEC (SPEEC) were studied for evaluation of the graft conditions. Group C had the most advanced preservation injury followed by group BIPC. GC count was lowest in group C, followed by BIPC. Comparison between groups BIPC and LIPC showed superior parameters (preservation injury, GC, EEC, and SPEEC) in LIPC. In conclusion, an IPC regimen of 2 ischemic cycles of 12 min each followed by 10 min of reperfusion distinctly decreased the preservation injury of intestinal grafts compared with non-manipulated grafts.
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http://dx.doi.org/10.1159/000327396DOI Listing
September 2011

Morphological and apoptotic changes in the intestinal mucosa and lung parenchyma after ischaemic/reperfusion injury of the jejunum.

Acta Vet Hung 2010 Jun;58(2):243-56

P. J. Safárik University, Department of Histology and Embryology, Faculty of Medicine, Srobárova 2, SK-04001 Kosice, Slovakia.

Ischaemic/reperfusion (IR) injury of the small intestine may lead to the development of multiple organ failure. Little is known about the morphological changes occurring in the organs during the subacute course of this syndrome. The objective of this study was to observe histopathological features and the role of apoptosis in the jejunal mucosa and lung parenchyma after intestinal IR injury in a long-term experiment. Wistar rats (n = 36) were divided into 4 experimental groups (IR(10), IR(20), IR(30), S). Groups IR(10), IR(20) and IR(30) (each n = 10) were subjected to 1-hour ischaemia of the cranial mesenteric artery followed by 10, 20 or 30 days of reperfusion, respectively. The control group S (n = 6) was not subjected to ischaemia. The jejunal mucosa remained intact after all periods of reperfusion. Apoptotic cells were found particularly in the lamina propria, with the most significant difference observed in the IR(30) group (P < 0.01). The lung parenchyma had lower regenerative capacity, which was confirmed by a high index of histological damage after 30 days of reperfusion (P < 0.01) and by the presence of an increased number of apoptotic cells, especially in the pulmonary interstitium. The number of apoptotic cells was ten times higher than in the control group (P < 0.001).
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http://dx.doi.org/10.1556/AVet.58.2010.2.10DOI Listing
June 2010

Development of jejunal graft damage during intestinal transplantation.

Ann Transplant 2009 Jul-Sep;14(3):62-9

Department of Histology and Embryology, Faculty of Medicine, P. J. Safárik University, Kosice, Slovakia.

Background: Intestinal transplantation (ITx) represents difficult life-saving intervention reserved for patients with irreversible intestinal failure. A serious complication of ITx is jejunal graft (JG) damage. The aim of the study was to evaluate the development of JG damage during ITx and determine the share of pathological elements (mechanical manipulation, ischemia, reperfusion) in this damage.

Material/methods: Male Wistar rats (n=60; 30 donors and 30 recipients) were used. The harvest of JG as well as heterotopic allotransplantation was performed using a technique adapted from Balaz et al. (2003). In all transplantations, three samples of JG were obtained: immediately after harvest (Sa1), after preservation (Sa2) and 60min after transplantation (Sa3). The samples were stained using the Hematoxylin&Eosin method and histopathological injury index (HII) was assessed using Park/Chiu classification. For detection and quantification of neuroendocrine cells (NECs) Singh's modification of the Masson-Hamperl argentaffin technique was used.

Results: The lowest level of HII was detected in Sa1=0.25+/-0.18; higher after preservation Sa2=1.42+/-0.38 and the highest HII was observed after transplantation Sa3=3.08+/-0.38. The percentage share of mechanical manipulation with the graft in jejunal damage during ITx was 8.11% (Sa1), the share of the ischemic element represented 37.98% (Sa2) and reperfusion had 53.91% of the share in jejunal damage (Sa3). The activity of NECs had sinusoidal character (Sa1=0.5+/-0.1; Sa2=1.4+/-0.0; Sa3=0.35+/-0.05).

Conclusions: The development of JG damage during ITx had progressive character. Mechanical manipulation had minimal influence on jejunal damage. One third of damage was caused by the ischemic component and the largest impact on JG damage resulted from reperfusion.
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November 2009

Nutritional depletion in relation to mortality in patients with chronic respiratory insufficiency treated with long-term oxygen therapy.

Wien Klin Wochenschr 2004 Sep;116(17-18):617-21

Department of Tuberculosis and Respiratory Medicine, Faculty of Medicine, P J Safárik University and L Pasteur Teaching Hospital, Cancer Institute, Kosice, Slovakia.

Objective: An association between malnutrition, weight loss and mortality has been demonstrated in patients with chronic obstructive pulmonary disease (COPD), but the prognostic influence of low body-mass index (BMI) and plasma concentrations of albumin and cholesterol is less clear in patients with chronic respiratory insufficiency treated with domiciliary long-term oxygen therapy (LTOT). We therefore analysed the prognostic value of BMI, plasma albumin and cholesterol concentrations in patients receiving LTOT.

Patients And Methods: From 1996 to 2001, LTOT was initiated in 255 patients. Analysis of the impact of nutritional status on survival was confined to a study group of 108 patients in whom the main outcomes, i.e. BMI, plasma cholesterol and albumin, were measured. Standard laboratory methods were used in the biochemical analyses. Pulmonary function was assessed with bodyplethysmography.

Results: 63 patients (58.3%) survived for two years post-initiation of LTOT and 45 patients (41.7%) did not. There were no differences between these two groups in pulmonary function tests and arterial blood gases at the start of LTOT. Overall, 10.2% of the study population were underweight, defined as BMI <20 kg/m2. Compared with patients who survived two years of LTOT, those who did not survive were older (69.3+/-0.9 versus 64.7+/-1.2 years, p<0.01), had significantly lower BMI (24.5+/-0.7 versus 26.5+/-0.7 kg/m2, p < 0.05), lower plasma cholesterol (4.61+/-0.19 versus 5.22+/-0.13 mmol/l, p<0.01) and lower plasma albumin concentrations (41.4+/-0.4 versus 42.8+/-0.4 g/l, p <0.05). Logistic regression analysis revealed that, in addition to age (p < 0.01), both BMI (p < 0.05) and cholesterol (p < 0.05) significantly affected the 2-year survival.

Conclusion: This study suggests that nutritional status is closely linked with prognosis in patients with chronic respiratory insufficiency treated with domiciliary LTOT: low BMI, low plasma cholesterol and low albumin are related to worse 2-year survival in such patients.
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http://dx.doi.org/10.1007/s00508-004-0226-6DOI Listing
September 2004