The Role of Endotoxaemia in the development of Renal Disorders Experimental Obstructive Jaundice in Rats

In rats with 2-week obstructive jaundice the sensitivity to endotoxin was studied and the effect of a single dose of endotoxin on histological development in the kidney, liver and spleen was also investigated. We were tested the effect on accumulation and distribution within organs, of fibrinogen labelled with radioactive iodine 125. We showed an increased sensitivity to endotoxin in obstructive jaundice. The cause of death in most rats was acute circulatory failure during the course of endotoxic shock, without clinical features of disseminated intravascular coagulation. In the isotope study, after endotoxin administration there was a specific dynamic increase of fibrinogen accumulation in the kidneys of rats with obstructive jaundice. We proposed, that the cause of the kidney changes during the course of obstructive jaundice could be the local activation of intrarenal coagulation

Transplantation of autologous bone marrow mononuclear cells with VEGF gene improves diabetic critical limb ischaemia

Wstęp: Celem pracy była ocena bezpieczeństwa i skuteczności skojarzonej terapii autogennymi jednojądrzastymi komórkami szpiku kostnego oraz terapii genowej plazmidem VEGF165 u chorych z krytycznym niedokrwieniem kończyn dolnych spowodowanym cukrzycą. Materiał i metody: U 16 chorych z bólami spoczynkowymi oraz niedokrwiennym owrzodzeniem kończyny dolnej w przebiegu cukrzycy zdecydowano o podaniu komórek jednojądrzastych i plazmidu VEGF165. Komórki jednojądrzaste szpiku oraz plazmid były podawane drogą iniekcji domięśniowych do mięśni niedokrwionej kończyny. Do oceny wyniku zastosowanej terapii określano poziom VEGF w surowicy oraz wskaźnik kostka-ramię. Do określenia stopnia odczuwania bólu została użyta skala wzrokowo-analogowa (VAS). Do wykazania w naczyniach wykonywano CT angiografię przed i po 3 miesiącach terapii. Wyniki: Średnie (&#177; SD) stężenie VEGF w osoczu wzrastało nieistotnie statystycznie z 257 &#177; 80 pg/l przed terapią do 391 &#177; 82 pg/l (P > 0,05) po 2 tygodniach od zakończenia leczenia. Wskaźnik kostka&#8211;ramię wzrósł istotnie statystycznie z poziomu 0,26 &#177; 0,22 przed terapią do 0,49 &#177; 0,30 (p < 0,001) po 3 miesiącach terapii. Zmniejszenie bólu spoczynkowego obserwowano u wszystkich pacjentów, średnia wartość VAS zmniejszyła się z 6,3 &#177; 1,4 przed terapią do 1,2 &#177; 1,1 po 3 miesiącach (p < 0,002). Angiogramy wykazały rozwój naczyń krążenia obocznego w 12 kończynach. Niedokrwienne owrzodzenie zostało całkowicie wyleczone w przepadku 12 chorych. Amputacje przeprowadzono tylko u 4 pacjentów z powodu zaawansowanego zakażenia rany, jakkolwiek poziom amputacji obniżono w tych przypadkach poniżej kolana. Powikłania były ograniczone do przemijających obrzęków podudzi u 2 chorych i gorączki u 2 pacjentów. Wniosek: Domięśniowa autotransplantacja komórek jednojądrzastych szpiku kostnego w połączeniu z podaniem phVEGF165 genu jest bezpieczną oraz skuteczną metodą leczenia pacjentów z cukrzycą i krytycznym niedokrwieniem kończyn dolnych. (Endokrynol Pol 2013; 64 (2): 129&#8211;138)Introduction: The aim of this study was to assess the safety and efficacy of combined autologous bone marrow mononuclear cell and VEGF165 gene therapy in patients with diabetes mellitus suffering from critical limb ischaemia (CLI). Material and methods: The administration of mononuclear cells (MNCs) and naked VEGF165 plasmid was performed in 16 limbs of 16 patients with rest pain and ischaemic ulcers due to diabetes. MNCs and plasmid were injected into the muscles of the ischaemic limbs. The levels of VEGF in serum and the ankle-brachial index (ABI) were measured before and after treatment. The Visual Analogue Scale (VAS) was used to evaluate pain sensation. CT angiography was performed before and after three months of therapy. Results: Mean (&#177; SD) plasma levels of VEGF increased non-significantly from 257 &#177; 80 pg/L to 391 &#177; 82 pg/L (p > 0.05) two weeks after therapy. The ABI improved significantly from 0.26 &#177; 0.22 to 0.49 &#177; 0.30 (p < 0.001) three months after therapy. A decrease in rest pain was observed in all patients; mean VAS decreased from 6.3 &#177; 1.4 to 1.2 &#177; 1.1 after three months (p < 0.002). Angiograms showed the development of collateral vessels in 12 limbs. Ischaemic ulcers healed in 12 limbs. Amputation was performed in four patients only, because of advanced wound infection. However, the level of amputations was lowered below knee level in these cases. Complications were limited to transient leg oedema in two patients and fever in two patients. Conclusions: Intramuscular bone marrow MNCs autotransplantation combined with the administration of phVEGF165 gene is safe, feasible and effective for patients with diabetes and CLI. (Endokrynol Pol 2013; 64 (2): 129&#8211;138

The use of the arterial homograft in treatment of local prosthetic graft infection - report of two cases

Infekcja po wszczepieniu protez z materiałów sztucznych stanowi ogromny problem w chirurgii naczyniowej. W pracy przedstawiono 2 przypadki zakażenia protezy naczyniowej aortalno-dwuudowej, ograniczone do dystalnej części ramienia. Ze względu na brak efektu leczenia przy zastosowaniu metod klasycznych chorych operowano, wymieniając zakażony odcinek na homograft tętniczy. Kontrola pooperacyjna wykazała ustąpienie infekcji.Infection after vascular operations, particularly after synthetic prosthetic graft implantation, remains a serious complication of reconstructive vascular surgery. In this paper there are presented two cases of bifurcated graft infection limited to the distal part of one of the branches. Because of the unsatisfactory results of conventional treatment, the patients underwent in situ replacement of the infected prosthesis with an arterial homograft. The postoperative check-up revealed the complete remission of infection

The Impairment of Wound Healing Process is Correlated With Abnormalities of TNF-α Production by Peritoneal Exudate Cells in Obstructive Jaundiced Rats

The wound healing process and production of tumour necrosis factor alpha (TNF-α) by peritoneal cells of 7-day and 14-day obstructive jaundice (OJ) and sham-operated rats were investigated. In the study the skin wound breaking strength was measured, In addition such histological and biochemical parameters as fibroblast and endothelial cell proliferation, inflammatory cell infiltration and hydroxyproline content were evaluated in polyurethane sponge discs implanted subcutaneously into rats. TNF-α production by peritoneal exudate cells (PEC), both spontaneous and lipopolysaccharide (LPS)- induced was determined by a bioassay. In OJ rats the process of both early as well as late phase of healing was impaired. The breaking strength of skin wound was decreased, the fibroblast and endothelial cell proliferation and collagen deposition, as well as hydroxyproline content were diminished. In 7 day OJ the numbers of inflammatory cells in the implants were lowered with a subsequent slight increase on day 14 of OJ. The spontaneous and LPS induced TNF- α production by PEC were significantly higher in 7 day OJ as compared with sham-operated controls. On day 14 of OJ the LPS-induced TNF-α level was, in contrast, much lower and did not differ much from the spontaneous TNF-α production. We conclude that the impairment of wound healing in OJ results from disturbances in functioning of the immune system caused by systemic endotoxaemia

Serum metabolomics approach to monitor the changes in metabolite profiles following renal transplantation

Systemic metabolic changes after renal transplantation reflect the key processes that are related to graft accommodation. In order to describe and better understand these changes, the 1HNMR based metabolomics approach was used. The changes of 47 metabolites in the serum samples of 19 individuals were interpreted over time with respect to their levels prior to transplantation. Considering the specific repeated measures design of the experiments, data analysis was mainly focused on the multiple analyses of variance (ANOVA) methods such as ANOVA simultaneous component analysis and ANOVA-target projection. We also propose here the combined use of ANOVA and classification and regression trees (ANOVA-CART) under the assumption that a small set of metabolites the binary splits on which may better describe the graft accommodation processes over time. This assumption is very important for developing a medical protocol for evaluating a patient’s health state. The results showed that besides creatinine, which is routinely used to monitor renal activity, the changes in levels of hippurate, mannitol and alanine may be associated with the changes in renal function during the post-transplantation recovery period. Specifically, the level of hippurate (or histidine) is more sensitive to any short-term changes in renal activity than creatinine

Clinical significance of oxidation and acetylation genetic polymorphism in patients with hyperthyreosis

Introduction: The relationship between genetically determined polymorphic oxidation and acetylation and susceptibility to some disease was aroused much interest. The aim of our study was to evaluate whether patients with hyperthyreosis differ from healthy persons in their ability to oxidize sparteine and acetylate sulphadimidine as model drugs. Oxidation and acetylation were estimated in 48 patients with hiperthyreosis. Material and methods: The control group consisted of 160 healthy volunteers for comparison of oxidation phenotype and 60 healthy volunteers for comparison of acetylation phenotype. The phenotyping of oxidation revealed two distinct populations among 40 patients with hyperthyreosis: 38 persons (95%) were extensive metabolizers (EM) of sparteine and 2 persons (5%) was poor metabolizers (PM). In 160 healthy persons (91.2%) were EM and 14 persons (8.8%) were PM. The difference between frequency distribution of PM and EM in healthy persons and in patients with hyperthyreosis was not statistically significant. Results: The phenotyping of acetylation showed among 48 patients with hyperthyreosis 8 persons (13%) were rapid acetylators (RA) and 40 persons (87%) were slow acetylators (SA). In 60 healthy volunteers the phenotype of rapid acetylation was observed in 31 persons (51%) and slow acetylation in 29 persons (49%). Relative risk (odds ratio) of development of thyroid diseases was 5.34 times higher for SA in comparison to RA. The prevalence of SA among patients with hyperthyreosis in comparison to healthy volunteers was statistically significant (p < 0.0002). Conclusions: The results of our study may suggest that slow acetylation phenotype is associated with increased risk of the development of hyperthyreosis.Wstęp: Badania nad udziałem czynników genetycznych w powstawaniu niektórych chorób prowadzi się w coraz szerszym zakresie. Celem pracy było stwierdzenie, czy między grupą chorych na nadczynność tarczycy a grupą kontrolną zdrowych ochotników istnieje różnica w zdolności utleniania sparteiny i acetylacji sulfadimidyny jako leków modelowych. Materiał i metody: Badaniami objęto 268 osób. W tej grupie 48 osób było chorych na nadczynność tarczycy. Grupę kontrolną dla fenotypu oksydacji stanowiło 160 zdrowych ochotników, natomiast dla fenotypu acetylacji 60 osób zdrowych. Wyniki: Wśród 40 chorych z nadczynnością tarczycy stwierdzono 38 (95%) ekstensywnych metabolizerów (EM, extensive metabolizers) i 2 (5%) słabych metabolizerów (PM, poor metabolizers) sparteiny. W grupie kontrolnej 160 ochotników, 146 (91,2%) okazało się EM, a 14 (8,8%) - PM. Niewielka przewaga częstości występowania PM wśród pacjentów z nadczynnością tarczycy w porównaniu z grupą osób zdrowych nie była statystycznie istotna. Wśród 48 chorych na nadczynność tarczycy wyróżniono 13% szybkich acetylatorów (RA, rapid acetylators) (8 osób) i 87% wolnych acetylatorów (SA, slow acetylators) (40 osób). Natomiast u 60 zdrowych ochotników stwierdzono 51% RA (31 osób) i 49% SA (29 osób). Odsetek szybkich i wolnych acetylatorów w grupie chorych z nadczynnością tarczycy różnił się w sposób statystycznie istotny w porównaniu z odsetkiem w grupie osób zdrowych (p < 0,0002). Względne ryzyko zachorowania na nadczynność tarczycy, wyrażone za pomocą proporcji szans (OR, odds ratio), jest ponad 5 razy większe dla wolnych niż dla szybkich acetylatorów. Wnioski: Wyniki niniejszych badań mogą sugerować, że osoby z fenotypem wolnej acetylacji są predysponowane do zapadalności na nadczynność tarczycy

Urine metabolome profiling of immune-mediated inflammatory diseases

Background: Immune-mediated inflammatory diseases (IMIDs) are a group of complex and prevalent diseases where disease diagnostic and activity monitoring is highly challenging. The determination of the metabolite profiles of biological samples is becoming a powerful approach to identify new biomarkers of clinical utility. In order to identify new metabolite biomarkers of diagnosis and disease activity, we have performed the first large-scale profiling of the urine metabolome of the six most prevalent IMIDs: rheumatoid arthritis, psoriatic arthritis, psoriasis, systemic lupus erythematosus, Crohn?s disease, and ulcerative colitis. Methods: Using nuclear magnetic resonance, we analyzed the urine metabolome in a discovery cohort of 1210 patients and 100 controls. Within each IMID, two patient subgroups were recruited representing extreme disease activity (very high vs. very low). Metabolite association analysis with disease diagnosis and disease activity was performed using multivariate linear regression in order to control for the effects of clinical, epidemiological, or technical variability. After multiple test correction, the most significant metabolite biomarkers were validated in an independent cohort of 1200 patients and 200 controls. Results: In the discovery cohort, we identified 28 significant associations between urine metabolite levels and disease diagnosis and three significant metabolite associations with disease activity (PFDR < 0.05). Using the validation cohort, we validated 26 of the diagnostic associations and all three metabolite associations with disease activity (PFDR < 0.05). Combining all diagnostic biomarkers using multivariate classifiers we obtained a good disease prediction accuracy in all IMIDs and particularly high in inflammatory bowel diseases. Several of the associated metabolites were found to be commonly altered in multiple IMIDs, some of which can be considered as hub biomarkers. The analysis of the metabolic reactions connecting the IMID-associated metabolites showed an overrepresentation of citric acid cycle, phenylalanine, and glycine-serine metabolism pathways. Conclusions: This study shows that urine is a source of biomarkers of clinical utility in IMIDs. We have found that IMIDs show similar metabolic changes, particularly between clinically similar diseases and we have found, for the first time, the presence of hub metabolites. These findings represent an important step in the development of more efficient and less invasive diagnostic and disease monitoring methods in IMIDs