Izolacija iz periferne krvi i analiza funkcija heterofila kokoši protočnom citometrijom – metodološko istraživanje

The aim of this study was to modify the flow cytometric method, which is used for analyzing neutrophils, to analyse the functions of avian heterophils. The blood samples used in the experiments were obtained from hens in slaughterhouses. Blood samples were collected from 10 hens for each trial. Within the scope of the present study, trials were carried out regarding the amount of blood, cell suspension, dihydrorodamine-123 (DHR-123), phorbol-12-myristate-13-acetate (PMA), and N-formyl-methionyl-leucyl-phenylalanine (fMLP), as well as the storage duration of blood samples and incubation time. The results showed that 0.5-3 ml of blood could be used to detect heterophil functions, and it would be ideal to conduct analyses using fresh blood samples. In addition, the results showed that blood stored at +4 °C for up to 8 hours may be also used if necessary. In order to isolate the cells, centrifugation for 30 minutes is sufficient, and it is appropriate to use a 30μL cell suspension. 2μL of DHR-123 should be used as a chemical probe to measure heterophil functions. Excessive use of DHR-123 affected the heterophil functions negatively. In addition, it was observed that using 2μL of fMLP, which is used as an oxidative burst stimulant, and 2μL of PMA as a stimulant of chemotaxic activity, were sufficient. It was concluded that incubation at 41 °C for 5 minutes after stimulating the heterophils is also sufficient. We conclude that the methods established in this study could be used to isolate heterophils and to analyze them by flow cytometry. Therefore, this study would contribute to further research and clinical studies in poultry.Cilj je istraživanja bio prilagoditi metodu protočne citometrije, koja se inače primjenjuje u analizi neutrofila, za analizu funkcija ptičjih heterofila. Uzorci krvi 10 kokoši dobiveni su iz klaonica. Uzevši u obzir količinu krvi, kao i trajanje pohrane krvnih uzoraka odnosno vrijeme inkubacije, analizirana su stanična suspenzija, dihidrorodamin-123 (DHR-123), forbol-12-miristat-13-acetat (PMA) i N-formil-metionil-leucil-fenilalanin (fMLP). Rezultati su pokazali da se 0,5 – 3 mL krvi može upotrijebiti za otkrivanje funkcija heterofila te da bi idealno bilo analizirati svježe uzorke krvi. Osim toga rezultati su pokazali da se i krv pohranjena na temperaturi od +4 °C, u vremenu do 8 sati, može upotrijebiti ako je to potrebno. Kako bi se stanice izolirale, dovoljno je centrifugirati 30 minuta uz primjenu stanične suspenzije od 30 μL. Kao kemijsku probu za mjerenje funkcija heterofila trebalo bi upotrijebiti 2 μL DHR-123. Prekomjerna upotreba DHR-123 negativno je utjecala na funkcije heterofila. Također, uočeno je da je bila dovoljna primjena 2 μL fMLP-a, koji služi kao stimulans oksidacijskog izgaranja, kao i primjena 2 μL PMA-a kao stimulansa kemotaktičke aktivnosti. Zaključeno je da je inkubacija na temperaturi od 41 °C, tijekom 5 minuta poslije stimulacije heterofila također dovoljna. Rezultati su pokazali da se metode ustanovljene u ovom radu mogu primijeniti za izolaciju heterofila i njihovu analizu protočnom citometrijom, čime se doprinosi daljnjim istraživanjima i kliničkim studijama u peradi

Contegra conduit for reconstruction of the right ventricular outflow tract: a review of published early and mid-time results

<p>Abstract</p> <p>Objective</p> <p>The valved conduit Contegra (bovine jugular vein) has being implanted for more than 7 years in the right ventricular outflow tract and it is noted that the available reports have been mixed. The aim of this study is to review the reported evidence in the literature.</p> <p>Methods</p> <p>Search of the relevant literature for the primary endpoints of operative mortality and morbidity and secondary endpoints of follow-up haemodynamic performance including severe stenosis, regurgitation and need for reintervention are presented.</p> <p>Results</p> <p>We selected and analysed 17 series including 767 patients. Commonest indication was Fallot's tetralogy. Operative mortality was 2.6%. Operative morbidity was 13.9%. In follow-up, the incidence of intraconduit stenosis was 10.9% (incidence of stenosis for the 12 millimetre conduit was 83.3% in one series) and that of at least moderate regurgitation was 6.3%.</p> <p>The aspirin users had a stenosis incidence of 10.5% compared to the non-users had a stenosis incidence of 9.6%.</p> <p>Conclusion</p> <p>A dissent on the performance of the Contegra is discussed, while results are satisfactory in the majority of studies apart for the smallest conduits (12 and 14 millimetre), suggesting an association to compromised run-off. The role of aspirin as antithrombotic modulator remains controversial.</p

Numerical Estimation of Film Thickness With an Air-Assisted Spray Gun

The main goal of this study is to present the effects of spraying parameters on the numerical evaluations of the fundamental behaviors of an air-assisted spray gun during the formation of child droplets in the spray flow field and material deposition on the target surface. For this purpose, first of all, the air-assisted spray gun geometry was created using the Solidworks software. Then, a computational domain with a 3D, unstructured grid structure was generated using the ANSYS-Workbench meshing tool. Numerical calculations were conducted using ANSYS-Fluent 2020-R2 commercial software. Different breakup models and their effects on the child droplet size were investigated. By coupling the Taylor analogy breakup (TAB) model and discrete phase model (DPM), the droplet size, trajectory, and coating thickness calculations were made under different atomizing air pressures. Also, the effects of spraying distance and droplet size on coating thickness and the critical Weber (We) number on the atomized particle diameter and particle speed were investigated. The results show that with the increase in atomizing air pressure, droplet sizes decrease and the film thickness on the center of the target surface and droplet speeds increases. Also, increasing the critical Weber number makes it more difficult to atomize the droplets

Cellulitis as a manifestation of miliary tuberculosis in a renal transplant recipient

Cutaneous involvement is an unusual presentation of tuberculosis (TB) and is rarely reported in renal transplant recipients. We describe a 37-year-old renal transplant recipient with disseminated Milcobacterium tuberculosis infection that presented as cellulitis. The organism was isolated from tissue and blood cultures. The patient was treated with quadruple anti-TB therapy for 12 months Anti-TB therapy led to a complete resolution of TB lesions We also provide a review of the literature on cutaneous TB in renal transplant recipients. Skin TB in renal transplant recipients usually occurs with nontuberculous mycobacteria. The spectrum of the skin lesions call be quite different and can mimic bacterial infections Mycobacteriosis should always be included in the differential diagnosis of a skin lesion in renal transplant recipients

Fever of unknown origin in uremic patients: A controlled prospective study

Twenty-nine uremic patients with fever of unknown origin(FUO) admitted to our clinic between 1994 and 1998 were evaluated prospectively. A group of 50 consecutive non-uremic patients with FUO followed up during the same period was used for comparison. The causes of FUO found in the uremic and non-uremic groups, respectively were as follows: infectious diseases, 69 vs. 44% (p=0.03); collagen vascular diseases, 6.9 vs. 6%; neoplasms, 3.4 vs. 26%; miscellaneous causes, 3.4 vs. 16%; and undiagnosed, 17.2 vs. 8%. Tuberculosis was the most common cause of FUO in both groups. The spectrum of underlying conditions for FUO in our uremic patients differed from that in the non-uremic patients and the uremic patients had a very high propensity for infectious diseases, especially tuberculosis