Expression of bone morphogenetic proteins in normal kidney and renal cell carcinoma

CILJ ISTRAŽIVANJA. Cilj doktorskog rada bio je prikazati izražaj koštanih morfogenetskih proteina (BMP), njihovih receptora (BMPR) i proteina njihovog signalnog puta (pSmad1/5/8) u zdravom tkivu bubrega, tkivu karcinoma bubrega (RCC) te tkivu na granici između karcinoma i normalnog bubrega. Nadalje, željeli smo prikazati promjenu u njihovoj ekspresiji između zdravog tkiva i RCC u usporedbi s kliničkim i patološkim značajkama ispitanika. ISPITANICI I METODE. Istraživanje je bilo prospektivno, a istražili smo tkivo bubrega u 64 ispitanika u kojih je, na Klinici za urologiju Kliničkog bolničkog centra Rijeka, učinjena nefrektomija zbog nemetastatskog karcinoma bubrega. Analizirali smo zdravo tkivo bubrega, tkivo karcinoma bubrega te granično tkivo, a analiza tkiva je provedena na Zavodu za anatomiju, Medicinskog fakulteta u Rijeci. Zabilježili smo ekspresiju proteina BMP-6, BMP-7, BMPR-IA, BMPR-IB, BMPR-II i pSmad1/5/8 (imunohistokemija i Western Blot: WB) odnosno njihove mRNA: mRNA BMP-2, mRNA BMP-4, mRNA BMP-6, mRNA BMP-7, mRNA BMPR-IA, mRNA BMPR-IB, BMP-RII (lančana reakcija polimeraze nakon reverzne transkripcije: RT-PCR). Razliku u ekspresiji BMP između različitih tkiva utvrdili smo korištenjem Student t testa (WB) i Mann-Whitney testa (RT-PCR). REZULTATI. U normalnom bubregu proteini BMP-7, BMPR-IA, BMPR-IB, BMPR-II i pSmad1/5/8 dominantno su bili prisutni u proksimalnim tubulima. U tkivu RCC pronašli smo ekspresiju proteina BMP-7, BMPR-IA, BMPR-IB, BMPR-II te pSmad 1/5/8. Ekspresiju BMP-6 u normalnom tkivu i u RCC nismo dokazali imunohistokemijskom metodom. Granično tkivo je pokazivalo obrazac ekspresije normalnog tkiva bubrega odnosno karcinoma bubrega u ovisnosti o kojem dijelu graničnog tkiva se radilo. Western blot metodom dokazali smo ekspresiju proteina BMP-6 i BMP-7, BMPR-IB, BMPR-II i pSmad1/5/8 u tkivu zdravog bubrega i u RCC. Ekspresija BMPR-IB i pSmad1/5/8 je bila povećana, a ekspresija BMPR-II, BMP-6 i BMP-7 je bila snižena u bolesnika s RCC. BMPR-IA se, ovom metodom, nije prikazao. mRNA svih BMP i njihovih receptora je pronađena u oba tkiva i bila je viša u RCC. Najsnažniji izražaj pokazao je mRNA BMP-2. Pronašli smo snažniju ekspresiju mRNA BMP-2 u bolesnika s višim tumorskim stadijem i višu ekspresiju mRNA BMP-6 u bolesnika starijih od 60 godina. Također je izražaj mRNA BMP-7 veći u žena. U uznapredovalom tumorskom stadiju (T3 i T4) bolesnika s RCC došlo je do gubitka ekspresije mRNA BMP-6, BMP-7 i BMPR-IB. ZAKLJUČAK. BMP i njihovi receptori su prisutni u zdravom tkivu bubrega te u tkivu karcinoma bubrega. BMP proteini, u ovoj studiji, su podjednakog izražaja u normalnom tkivu i karcinomu bubrega ali je ekspresija njihove mRNA znatno veća u RCC. Spoznaja kako je mRNA BMP-2 snažno eksprimiran u RCC te kako njegova ekspresija ovisi o tumorskom stadiju govori u prilog njegove važnosti u tumorigenezi karcinoma bubrega koju bi trebalo daljnjim istraživanjima ispitati.OBJECTIVES. Aim of our study was to present expression of different bone morphogenetic proteins (BMPs), their receptors (BMPRs) and protein of their signaling pathway (pSmad1/5/8) in healthy kidney tissue, renal cell carcinoma (RCC) and borderline tissue. Also, we compared changes in expression between healthy kidney tissue and RCC and correlate their expression with clinicopathological parameters. PATIENTS AND METHODS. We prospectively examined kidney tissue samples of 64 patients who underwent nephrectomy for localized kidney cancer. All patients were operated in Department of Urology, University Hospital Rijeka. Analysis was performed on healthy kidney tissue, RCC and borderline tissue on Institute of Anatomy, School of Medicine Rijeka. Protein expression of BMP-6, BMP-7, BMPR-IA, BMPR-IB, BMPR-II and pSmad 1/5/8 was determinated with immunohistochemistry and Western blot:WB) and expression of mRNA BMP-2, mRNA BMP-4, mRNA BMP-6, mRNA BMP-7, mRNA BMPR-IA, mRNA BMPR-IB, BMP-RII with reverse transcriptase polymerase chain reaction (RT-PCR). Difference in expression between tissue was estimated by Student t test (WB) and Mann-Whitney test (RT-PCR). RESULTS. In healthy kidney proteins BMP-7, BMPR-IA, BMPR-IB, BMPR-II and pSmad1/5/8 were dominantly expressed in proximal tubule. In RCC expression of proteins BMP-7, BMPR-IA, BMPR-IB, BMPR-II and pSmad 1/5/8 was found. Expression of BMP-6 was not noticed in any tissue with immunohistochemistry. Borderline tissue showed expression like RCC or healthy tissue in dependence which part of borderline tissue was examined. Proteins BMP-6, BMP-7, BMPR-IB, BMPR-II and pSmad1/5/8 were detected with WB in RCC and normal kidney. Expression of BMPR-IB and pSmad1/5/8 was increased and expression of BMPR-II, BMP-6 and BMP-7 was decreased in RCC. BMPR-IA was not detected with WB. mRNA of all BMPs and BMPRs was detected in both tissue with higher expression in RCC, specially for BMP-2. Expression of mRNA BMP-2 was increased in patients with higher tumor stage, mRNA BMP-6 was increased in elderly (> 60 years) and mRNA BMP-7 in female. We also found loss of mRNA BMP-6, BMP-7 i BMPR-IB expression in tumor stage T3 and T4. CONCLUSION. BMPs and their receptors was expressed in healthy kidney tissue and RCC. BMP proteins, in our study, have similar level of expression in both tissue but expression of their mRNA was higher in RCC. High level of mRNA BMP-2 expression in RCC and their increased expression with higher tumor stage are arguments in favour of their role in RCC. Futher investigation is needed to elucidate role of BMP-2 in RCC

Urinary bladder catheterization – modern approach

Kateterizacija je rutinski postupak kojim se omogućava drenaža urina iz mokraćnoga mjehura. Može se učiniti u dijagnostičke i terapijske svrhe. Postoji mnogo različitih katetera, a njihov izbor ovisi o razlogu njegova postavljanja. Kateteri za jednokratnu kateterizaciju obično su ravni, napravljeni od polivinil-klorida ili, ponekad, od lateksa. Trajni kateteri imaju retencijski mehanizam, a najčešće se koristi Foleyjev kateter (balon služi kao retencijski mehanizam). Kod dugotrajne kateterizacije preporučuje se korištenje Foleyjeva katetera od biokompatibilnog materijala. Silikonski kateteri, kao i oni obloženi hidrogelom, bolje se toleriraju nego oni proizvedeni od lateksa ili poliuretana. Potrebno je izabrati urinarni kateter najmanjega promjera, koji zadovoljava svrhu kateterizacije. Duljina kateterizacije mora biti što je kraće moguća. Otežana kateterizacija najčešće je prisutna kod muškaraca. Njezini uzroci mogu biti striktura uretre, uvećana prostata ili skleroza vrata mjehura. Otežana kateterizacija u žena prisutna je kod adipoznih osoba ili u slučaju kada se ne može pronaći vanjsko ušće uretre. Najčešća komplikacija vezana uz kateterizaciju je infekcija mokraćnoga sustava, koja sa sobom donosi značajan i morbiditet i mortalitet. To je inače i najčešći uzrok nozokomijalnih infekcija, čak do 40 %. Ostale moguće komplikacije su parafimoza, „lažni prolaz”, striktura uretre, perforacija uretre i krvarenje. Kateterizacija mokraćnoga mjehura u većine je pacijenata jednostavan postupak koji zbog mogućih popratnih komplikacija zahtijeva standardiziran pristup i provedbu.Catheterization is a routine medical procedure that facilitates direct drainage of the urinary bladder. It is used for both the diagnostic and therapeutic purposes. Many types of catheters are available for urethral catheterization, and the choice of a specific type of catheter depends on the reason for catheterization. The catheter for one-time catheterization is usually a straight catheter made from polyvynyl chloride or sometimes latex. For long-term catheterization catheters need a retention mechanism. Foley type catheters (balloon served as retention mechanism) are most often used for long-term urethral catheterization. If long-term catheterization is anticipated, it is advisable to use a Foley catheter made of the most biocompatible material. Catheters made of silicone or coated with hydrogel are, in general, better tolerated over the long-term than those made of materials like latex and polyurethane. In addition, one should choose the smallest urethral catheter that will accomplish the purpose of catheterization. The duration of catheterization must be as short as possible. Difficult catheterization is mostly seen in male patients. The most frequents causes are urethral stricture, prostatic enlargement and bladder neck contracture. Difficulty in catheterization of the female urethra is uncommon and usually results from extreme obesity and inability to locate the urethral meatus. The most frequent complication associated with catheterization is catheter-associated urinary tract infection resulting in significant morbidity and mortality. It is the most common nosocomial infection, comprising >40 % of all institutionally aquired infections. Other possible complications are paraphimosis, creation of false passage, urethral stricture, urethral perforation and bleeding. Catheterization is a routine procedure in the majority of patients but because of potential complications, performance must be standardized

Health care of patients suffering from autonomic dysreflexia

Autonomna disrefleksija (AD) klinički je sindrom koji se javlja u osoba sa znatnim oštećenjem kralježnične moždine (OKM). AD se najčešće pojavljuje u osoba s OKM-om u visini Th6 ili više, ali može nastati i u osoba s OKM-om do Th10. Uzrok nastanka sindroma je neinhibirani odgovor autonomnog živčanog sustava na neki štetan podražaj (primjerice, retenciju urina ili konstipaciju) nastao ispod razine ozljede. Radi se o sindromu koja zahtijeva brzo prepoznavanje i promptno liječenje. Neliječena AD može uzrokovati cerebrovaskularne i kardiovaskularne komplikacije i smrtni ishod, a sve kao posljedica nekontrolirane hipertenzije. Prevencija AD-a je posebno važna. Preventivne metode moraju poznavati osobe s OKM-om, članovi obitelji i zdravstveni djelatnici. Zdravstveni djelatnici trebali bi educirati bolesnike i članove njihove obitelji ili osobe koje se brinu za njih, a sve kako bi se AD prepoznala na vrijeme i spriječio nastanak komplikacija koje su opasne za život. Edukacijski programi trebaju sadržavati usvajanje preventivnih strategija, prepoznavanje znakova i simptoma AD-a te pravilno liječenje sindroma. Medicinske sestre/tehničari najmnogobrojnija su skupina zdravstvenih djelatnika i ponajviše su u izravnom kontaktu s pacijentima, stoga je njihova uloga u prevenciji, prepoznavanju simptoma autonomne disrefleksije i u edukaciji važna za poboljšanje stupnja kvalitete života osoba s OKM-om

Surgical Technique in the Rat Model of Kidney Transplantation

Today successful kidney transplantation procedures, techniques and immunosupression protocols are a consequence of extensive research on animal models. During every transplantation surgery there are two crucial points for the success of the entire procedure: vascular (arterial end venous) and ureteral or ureterovesical anastomosis. Renal artery and vein of the donor kidney can be anastomosed end-to-side to the abdominal aorta and vena cava of the recipient (heterotopic transplantation), or end-to-end to the remains of renal artery and vain of the recipient (orthotopic transplantation) after nepherctomy. The ureter can be anastomosed also end-to-en or we can connect it directly to the urinary bladder (ureterocystoneostomy). The aim of this study was to elucidate which technique has better results according to: animal survival, reperfusion and perfusion of the transplanted kidney, elimination of the urine from the transplanted kidney and procedure costs. The study included 240 (120 donors and 120 recipients) male Wistar rats (3 months old; weight 250–300 g Our results are clearly showing that the end-to-end vascular anastomosis, and Paquins ureterovesical anastomosis have better results in transplanted rat kidneys survival and urine drainage compared to end-to-side vascular anastomosis and end-to-end ureteral anastomosis. Based on our experience we can conclude that described methods of end-to-end vascular anastomosis and Paquins ureterovesical anastomosis are less technically demanding and have a shorter learning curve. Therefore, we can recommend the use of described methods in kidney transplantation related researches

UROLITHIASIS – PREVENTION, DIAGNOSIS AND TREATMENT

Broj bolesnika s urolitijazom sve je veći. Većina kamenaca nastaje kao posljedica metaboličke abnormalnosti, ali, unatoč tome, u većine pacijenata ne može se pronaći specifična metabolička abnormalnost. Stoga u većini slučajeva nije mogu}e kauzalno liječenje već samo simptomatska terapija. Iako većina bolesnika izmokri kamence, u preostalih se koriste različite, prije svega, kirurške metode uklanjanja kamenca. U prevenciji kamenaca najvažniji su čimbenici velik unos tekućine, uz posljedičnu obilnu diurezu, te dijetalne mjere.The prevalence of urinary stones is increasing. The majority of stones are caused by metabolic abnormality but, despite this, specific metabolic abnormality cannot be found in most patients. In most cases causal therapy is not possible so that the therapy of symptoms is the only option. Despite the fact that most patients spontaneously eliminate stones, different surgical methods are available. The cornerstone of stone prevention is a high fluid input with increased urine volume as well as diet

Expression of Bone Morphogenetic Protein-7, It´s Receptors and Smad1/5/8 in Normal Human Kidney and Renal Cell Cancer

Bone morphogenetic proteins (BMPs) are cytokines which are important for kidney homeostasis but also have role in the some renal diseases and renal cell carcinoma (RCC)1–5. In the last three decades incidence of RCC was constantly increased and the role of different molecular biomarkers in RCC is explored1. We analyzed expression of BMP-7, their receptors (BMPR-IA, BMPR-IB, BMPR-II) and proteins of their signaling pathway (pSmad1/5/8) in sixteen renal cancer samples and paired normal tissue. Tissue samples were analyzed by immunohistochemistry and Western blot. BMP-7, BMP receptors and pSmad1/5/8 were expressed in all structures of normal kidney but dominantly in the proximal tubular cells. In the cancer samples their expression was also noticed. Comparison of BMPs between different tissue showed increased expression of BMPR-IB and pSmad 1/5/8 and decreased expression of BMP-7 and BMPR-II in RCC compared to normal kidney. BMPR-IA was detected with immunohistochemistry but with Western blot attenuated signal was presented. BMP-7, BMP receptors and pSmad1/5/8 were showed in normal kidney and RCC. Detected alterations of BMP-7, BMP receptors and pSmad expression in RCC suggested their possible role in tumorigenesis of kidney cancer

UROLITHIASIS – PREVENTION, DIAGNOSIS AND TREATMENT

Broj bolesnika s urolitijazom sve je veći. Većina kamenaca nastaje kao posljedica metaboličke abnormalnosti, ali, unatoč tome, u većine pacijenata ne može se pronaći specifična metabolička abnormalnost. Stoga u većini slučajeva nije mogu}e kauzalno liječenje već samo simptomatska terapija. Iako većina bolesnika izmokri kamence, u preostalih se koriste različite, prije svega, kirurške metode uklanjanja kamenca. U prevenciji kamenaca najvažniji su čimbenici velik unos tekućine, uz posljedičnu obilnu diurezu, te dijetalne mjere.The prevalence of urinary stones is increasing. The majority of stones are caused by metabolic abnormality but, despite this, specific metabolic abnormality cannot be found in most patients. In most cases causal therapy is not possible so that the therapy of symptoms is the only option. Despite the fact that most patients spontaneously eliminate stones, different surgical methods are available. The cornerstone of stone prevention is a high fluid input with increased urine volume as well as diet