Uropathogenic Escherichia coli Induces Serum Amyloid A in Mice following Urinary Tract and Systemic Inoculation

Serum amyloid A (SAA) is an acute phase protein involved in the homeostasis of inflammatory responses and appears to be a vital host defense component with protective anti-infective properties. SAA expression remains poorly defined in many tissues, including the urinary tract which often faces bacterial challenge. Urinary tract infections (UTIs) are usually caused by strains of uropathogenic Escherichia coli (UPEC) and frequently occur among otherwise healthy individuals, many of whom experience bouts of recurrent and relapsing infections despite the use of antibiotics. To date, whether SAA is present in the infected urothelium and whether or not the induction of SAA can protect the host against UPEC is unclear. Here we show, using mouse models coupled with immunofluorescence microscopy and quantitative RT-PCR, that delivery of UPEC either directly into the urinary tract via catheterization or systemically via intraperitoneal injection triggers the expression of SAA. As measured by ELISA, serum levels of SAA1/2 were also transiently elevated in response to UTI, but circulating SAA3 levels were only up-regulated substantially following intraperitoneal inoculation of UPEC. In in vitro assays, physiological relevant levels of SAA1/2 did not affect the growth or viability of UPEC, but were able to block biofilm formation by the uropathogens. We suggest that SAA functions as a critical host defense against UTIs, preventing the formation of biofilms both upon and within the urothelium and possibly providing clinicians with a sensitive serological marker for UTI

Therapist\u27s experience of psychotherapy relationship through the lens of attachment theory

V pričujočem delu se avtorica loteva raziskovanja terapevtovega doživljanja terapevtskega odnosa in sicer skozi prizmo navezanosti. Teoretični uvod tako predstavlja podrobno predstavitev teorije navezanosti od Bowlbyja naprej. Avtorica pregleda zgodovino koncepta kot tudi njegov razvoj in etiologijo. Drugi del teoretičnega uvoda je sestavljen iz predstavitve koncepta terapevtskega delovnega odnosa in njegove implikacije za terapevtsko delo. Avtorica se znova poglobi v razumevanje koncepta in skuša slediti njegovemu razvoju. Delo nato preide na raziskovalni del. Avtorico je zanimalo, kako terapevti doživljajo terapevtski odnos skozi lečo navezanosti. Za ta namen je terapevtski odnos razdelila na tri faze – fazo prednavezanosti, ki jo zaznamuje vzpostavljanje kontakta in učenje pravil terapevtskega dela, sledi ji faza oblikovanja navezanosti, kjer je dinamika že utečena, klienti pa opažajo prve spremembe v svojem življenju, in zadnja faza, torej faza navezanosti, ki jo oblikuje visoka stopnja zaupanja, jasna komunikacija med klientom in terapevtom in dobra delovna aliansa. Avtorica je za namen naloge intervjuvala osem terapevtov in podatke, pridobljene v teh intervjujih analizirala s pomočjo kvalitativne metode fenomenološke raziskave. Ugotovitve so pokazale, da se doživljanje terapevtov v veliki meri sklada z do sedaj aktualno teorijo. V fazi prednavezanosti so terapevti poročali o tem, da se odnos šele oblikuje, zaupanje vzpostavlja, komunikacija še usklajuje. V fazi oblikovanja navezanosti terapevti poročajo o naraščajočem zaupanju in opažanju sprememb in v zadnji fazi poročajo o močni navezanosti in z rastjo odnosa vedno bolj enakovreden odnos med terapevtom in klientom. To je tudi obdobje, ko klient in terapevt načrtujeta zaključek. Raziskovalno delo je avtorici ponudilo vpogled v globoko doživljanje terapevtskih odnosov s strani terapevta, spoznala je izzive, s katerimi se terapevti srečujejo pri svojem delu, pa tudi dobrobiti terapevtskega dela. V svojem delu je avtorica izpostavila tudi pomanjkanje informacij s področja raziskovanja terapevtske perspektive.In the current master thesis, the author is exploring a therapists’ perception of psychotherapy process through the lens of attachment. The theoretical part consists of precise description of ontogeny of the attachment theory from his beginner, John Bowlby. The author follows the history and etiology of the mentioned concept. The other part of the master thesis consists of presentation of the working alliance concept and its indication for therapeutic work. Author tries to get an overall understanding about the concept, its origins and current implications. In the current master thesis, the author was interested in therapists’ perception of psychotherapy relationship through the attachment perspective. To enquire about it she divided the attachment process on three main stages – the preattachment stage, characterized by first contact and establishment of communication, second stage, namely attachment in the making stage, where the trust and therapy framework is already established and clients can see first changes, and last, third phase that is the attachment phase, where therapist and clients have high level of trust, good communication and good working alliance. Author interviewed eight therapists and analyzed the information from the interviews via phenomenological research. Results have shown that the way therapists from the research see their relationships folds well with the current theory from the field. In the preattachment stage the therapist spoke about the way the relationship has yet to be built, the trust has yet to be gained and communication is still on shaky grounds. In the second, attachment in the making phase, therapists observe deeper trust, clients start seeing changes in their lives and in the last, attachment phase they describe the relationship as of one of strong attachment. With continuation of the relationship the relationship itself become more of an equal one and is, as such, slowly transcending into the conclusion phase. The research work enabled the author to see the depth of the therapists’ experiencing of the relationships they make with their clients, she get to know the challenges of the therapeutic relationships and also the benefits of it. As a conclusion, the author exposes the lack of information available on research done of exploring the therapists’ perspective

Poly-L-lysine as an effective and safe desquamation inducer of urinary bladder epithelium

Induced desquamation of urinary bladder epithelial cells, also called urothelial cells, is frequently used in studies of bladder epithelial regeneration and also in treating recurrent bacterial cystitis. Positively charged polymer chitosan is known to cause large-scale desquamation of terminally differentiated urothelial cells called umbrella cells. Aiming to compare the desquamation ability of another polycation poly-L-lysine, we studied the effect of this polymer on the functional and structural integrity of the urothelium in ex vivo and in vivo experiments. The urothelium was analyzed by measuring transepithelial electrical resistance, and the structural changes of its luminal surface were analyzed with scanning electron microscopy. The results revealed a selective and concentration-dependent desquamation effect of poly-L-lysine on superficial urothelial cells followed by quick regeneration of the urothelium, which functionally and structurally recovers in 2 to 3 h after poly-L-lysine-induced injury. Poly-L-lysine was thus proven to be a promising polymer to be used when desquamation of urothelial cells is required in basic and potentially clinical studies

The Use of Polymer Chitosan in Intravesical Treatment of Urinary Bladder Cancer and Infections

The most frequent diseases of the urinary bladder are bacterial infections and bladder cancers. For both diseases, very high recurrence rates are characteristic: 50–80% for bladder cancer and more than 50% for bladder infections, causing loss of millions of dollars per year for medical treatment and sick leave. Despite years of searching for better treatment, the prevalence of bladder infections and bladder cancer remains unchanged and is even increasing in recent years. Very encouraging results in treatment of both diseases recently culminated from studies combining biopolymer chitosan with immunotherapy, and chitosan with antibiotics for treatment of bladder cancer and cystitis, respectably. In both pathways of research, the discoveries involving chitosan reached a successful long-lasting cure. The property of chitosan that boosted the effectivity of illness-specific drugs is its ability to enhance the accessibility of these drugs to the very sources of both pathologies that individual treatments without chitosan failed to achieve. Chitosan can thus be recognised as a very promising co-player in treatment of bladder cancer and bacterial cystitis

The use of polymer chitosan in intravesical treatment of urinary bladder cancer and infections

The most frequent diseases of the urinary bladder are bacterial infections and bladder cancers. For both diseases, very high recurrence rates are characteristic: 50-80% for bladder cancer and more than 50% for bladder infections, causing loss of millions of dollars per year for medical treatment and sick leave. Despite years of searching for better treatment, the prevalence of bladder infections and bladder cancer remains unchanged and is even increasing in recent years. Very encouraging results in treatment of both diseases recently culminated from studies combining biopolymer chitosan with immunotherapy, and chitosan with antibiotics for treatment of bladder cancer and cystitis, respectably. In both pathways of research, the discoveries involving chitosan reached a successful long-lasting cure. The property of chitosan that boosted the effectivity of illness-specific drugs is its ability to enhance the accessibility of these drugs to the very sources of both pathologies that individual treatments without chitosan failed to achieve. Chitosan can thus be recognised as a very promising co-player in treatment of bladder cancer and bacterial cystitis

Does the urothelium of old mice regenerate after chitosan injury as quickly as the urothelium of young mice?

The aging of organisms leads to a decreased ability of tissue to regenerate after injury. The regeneration of the bladder urothelium after induced desquamation with biopolymer chitosan has been studied in young mice but not in old mice. Chitosan is a suitable inducer of urothelial desquamation because it is known to be non-toxic. We used chitosan for desquamation of urothelial cells in order to compare the dynamics of urothelial regeneration after injury between young and old mice. Our aim was to determine whether the urothelial function and structure of old mice is restored as fast as in young mice, and to evaluate the inflammatory response due to chitosan treatment. We discovered that the urothelial function restored comparably fast in both age groups and that the urothelium of young and old mice recovered within 5 days after injury, although the onset of proliferation and differentiation appeared later in old mice. Acute inflammation markers showed some differences in the inflammatory response in young versus old mice, but in both age groups, chitosan caused short-term acute inflammation. In conclusion, the restoration of urothelial function is not impaired in old mice, but the regeneration of the urothelial structure in old mice slightly lags behind the regeneration in young mice

Poly-L-lysine as an Effective and Safe Desquamation Inducer of Urinary Bladder Epithelium

Induced desquamation of urinary bladder epithelial cells, also called urothelial cells, is frequently used in studies of bladder epithelial regeneration and also in treating recurrent bacterial cystitis. Positively charged polymer chitosan is known to cause large-scale desquamation of terminally differentiated urothelial cells called umbrella cells. Aiming to compare the desquamation ability of another polycation poly-L-lysine, we studied the effect of this polymer on the functional and structural integrity of the urothelium in ex vivo and in vivo experiments. The urothelium was analyzed by measuring transepithelial electrical resistance, and the structural changes of its luminal surface were analyzed with scanning electron microscopy. The results revealed a selective and concentration-dependent desquamation effect of poly-L-lysine on superficial urothelial cells followed by quick regeneration of the urothelium, which functionally and structurally recovers in 2 to 3 h after poly-L-lysine–induced injury. Poly-L-lysine was thus proven to be a promising polymer to be used when desquamation of urothelial cells is required in basic and potentially clinical studies

Lectins as Biomarkers of IC/BPS Disease: A Comparative Study of Glycosylation Patterns in Human Pathologic Urothelium and IC/BPS Experimental Models

Pathophysiology of interstitial cystitis/bladder pain syndrome (IC/BPS) remains poorly understood, as well as its effective diagnosis and therapy. Studying changes in tissue glycosylation patterns under pathological conditions is a promising way of discovering novel biomarkers and therapeutic targets. The glycobiology of IC/BPS is largely understudied, therefore we compared glycosylation patterns of normal human urothelium with the urothelium of IC/BPS patients using a selection of 10 plant-based lectins with different monosaccharide preferences. We also compared lectin binding to human urothelium with the two most cited experimental models of IC/BPS, specifically, TNFα-treated human urothelial cell line RT4 and cyclophosphamide-induced chronic cystitis in C57BL6/J mice. Furthermore, binding of four of the selected lectins (ConA, DSL, Jacalin and WGA) was evaluated qualitatively by means of fluorescence microscopy, and quantitatively by fluorescence intensity (F.I.) measurements. Our results reveal a significant reduction in F.I. of Jacalin, as well as a prominent change in the WGA labeling pattern in the urothelium of IC/BPS patients, suggesting their potential use as promising additional biomarkers for histopathological diagnosis of IC/BPS. We have also shown that urothelial glycosylation patterns between selected experimental models and patients with IC/BPS are similar enough to offer an adequate platform for preclinical study of IC/BPS glycobiology