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Although, liver transplantation serves as the only curative treatment for patients with end- stage liver diseases, it is burdened with complications, which affect survival rates. In addition to clinical risk factors, contribution of recipient and donor genetic prognostic markers has been extensively studied in order to reduce the burden and improve the outcomes. Determination of single nucleotide polymorphisms (SNPs) is one of the most important tools in development of personalized transplant approach. To provide a better insight in recent developments, we review the studies published in the last three years that investigated an association of recipient or donor SNPs with most common issues in liver transplantation: Acute cellular rejection, development of new-onset diabetes mellitus and non-alcoholic fatty liver disease, hepatocellular carcinoma recurrence, and tacrolimus concentration variability. Reviewed studies confirmed previously established SNP prognostic factors, such as PNPLA3 rs738409 for non-alcoholic fatty liver disease development, or the role of CYP3A5 rs776746 in tacrolimus concentration variability. They also identified several novel SNPs, with a reasonably strong association, which have the potential to become useful predictors of post-transplant complications. However, as the studies were typically conducted in one center on relatively low-to-moderate number of patients, verification of the results in other centers is warranted to resolve these limitations. Furthermore, of 29 reviewed studies, 28 used gene candidate approach and only one implemented a genome wide association approach. Genome wide association multicentric studies are needed to facilitate the development of personalized transplant medicine.

Ferritin is the main intracellular storage of iron. Animal studies show that female liver and kidney express more ferritin and accumulate more iron than male. However, no study so far has investigated sex and age differences in light (FtL) and heavy (FtH) ferritin chain expression. To address this, we relied on specific antibodies and immunochemical methods to analyse the expression of both ferritin chains in the liver and kidney of 3-month and 2-year-old male and female Wistar rats. To see how sex hormones may affect expression we also studied adult animals gonadectomised at the age of 10 weeks. FtL and FtH were more expressed in both organs of female rats, while gonadectomy increased the expression in males and decreased it in females, which suggests that it is stimulated by female and inhibited by male steroid hormones. Normal kidney ferritin distribution and change with aging warrant more attention in studies of (patho) physiological and toxicological processes., Feritin je glavni skladišni protein unutarstaničnoga željeza. U korelaciji s ekspresijom feritina, istraživanja na životinjama pokazuju da jetra i bubrezi u ženki nakupljaju više željeza nego u mužjaka. Međutim, dosad se nijedno istraživanje nije bavilo spolnim i dobnim razlikama u ekspresiji lakog (FtL) i teškog (FtH) lanca koji čine feritinski nanokavez. Kako bismo to riješili, oslonili smo se na specifična protutijela i imunokemijske metode za analizu ekspresije obaju feritinskih lanaca u jetri i bubrezima odraslih i starih mužjaka i ženki štakora soja Wistar. Da bismo vidjeli kako spolni hormoni mogu utjecati na ekspresiju feritina, proučavali smo i gonadektomirane odrasle životinje. FtL i FtH bili su izraženiji u odraslih i starih životinja u obama organima u ženki štakora; gonadektomija je povećala ekspresiju u mužjaka, a smanjila je u ženki, što upućuje na to da feritine stimuliraju ženski, a inhibiraju muški steroidni hormoni. Normalna raspodjela feritina u bubrezima i promjena sa starenjem zahtijevaju više pažnje u proučavanju (pato)fizioloških i toksikoloških procesa.

International audience; Background: Silver nanoparticles (AgNP) are one of the most commercialized types of nanomaterials, with a wide range of applications owing to their antimicrobial activity. They are particularly important in hospitals and other healthcare settings, where they are used to maintain sterility of surfaces, textiles, catheters, medical implants, and more. However, AgNP can not only harm bacteria, but also damage mammalian cells and tissue. While the potential toxicity of AgNP is an understood risk, there is a lack of data on their toxicity in combination with polymeric materials, especially plastic nanoparticles such as polystyrene nanoparticles (PSNP) that can be released from surfaces of polystyrene devices during their medical use. Aim: This study aimed to investigate combined effect of AgNP and nanoplastics on human immune response. Methods: Cells were treated with a range of PSNP and AgNP concentrations, either applied alone or in combination. Cytotoxicity, induction of apoptosis, generation of oxidative stress, uptake efficiency, intracellular localization and nanomechanical cell properties were selected as exposure biomarkers. Results: Collected experimental data showed that nanomixture induced oxidative stress, apoptosis and mortality of Jurkat cells stronger than its individual components. Cell treatment with AgNP/PSNP mixture also significantly changed cell mechanical properties, evidenced by reduction of cells' Young Modulus. Conclusion: AgNP and PSNP showed additive toxic effects on immortalized human lymphocytes, evidenced by increase in cellular oxidative stress, induction of apoptosis, and reduction of cell stiffness. These results have important implications for using AgNP and PSNP in medical contexts, particularly for long-term medical implants.

The current “gold standard” therapy is levodopa (L-DOPA) due to its effectiveness and ability to permeate the blood-brain barrier (BBB). However, long-term treatment with L-DOPA can lead to serious side effects and increase oxidative stress, thereby worsening the disease [2]. The oxidative stress caused by either L-dopa or dopamine could be lessened by the combination of antioxidants – one that promises is selenium (Se) – an essential trace element that can protect from oxidative damage through selenoproteins [3]. Direct use of Se as an antioxidant supplement is toxic due to a narrow therapeutic index, so Se should be applied in other therapeutical forms. The use of Se in the form of nanoparticles (SeNPs) may elicit the same beneficial effects with reduced toxicity [4]. This study tested the safety of SeNPs as L-DOPA carrier using differentiated neuroblastoma (SH- SY5Y) cells as in vitro model of Parkinson`s disease [5]. SeNPs were prepared via reduction of sodium selenite by L-ascorbic acid and functionalization by polysorbate 20 (SeTWEEN) and poly(vinylpyrrolidone) (SePVP). The shape, size, and surface charge of SeNPs were determined with transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta potential measurements, respectively. Cell viability and apoptosis induction test were performed using flow cytometry, while oxidative stress was determined by measuring the concentration od reactive oxygen species, glutathione (GSH) and mitochondrial membrane potential. In all experiments L-DOPA loaded SeNPs were compared with SeNPs and L-DOPA alone. L- DOPA loaded SeNPs (1ppm) were non-toxic to differentiated SH-SY5Y cells after 24-hour treatment, while L-DOPA reduced cell viability at the 100 μM, but not at 50 μM concentration. L-DOPA induced oxidative stress at the concentration of 100 µM which was effectively reduced after combining with SeNPs (1 ppm). Results suggest that SeNPs could be a promising approach for the reduction of toxicity caused by L-DOPA.

Low-grade squamous intraepithelial lesion (SIL) is a cytologic diagnosis etiologically related to human papilloma virus (HPV) infection that leads to the release of inflammation mediators, the formation of reactive oxygen species (ROS) and decreased levels of antioxidants in tissues, which is why antioxidants might be considered effective against SIL progression. This randomized double-blind placebo-controlled study aimed to investigate the effectiveness of alpha-lipoic acid (ALA) supplementation (600 mg/day) on the regression of low-grade SIL in 100 patients. Low-grade SIL was determined after the cytological screening, colposcopic examination and targeted biopsy and histological confirmation of cytological–colposcopic diagnosis. Inflammation parameters and the presence of HPV were determined by standard laboratory methods. Dietary and lifestyle habits were investigated using a standardized and validated semi-quantitative food questionnaire (FFQ). ALA supplementation significantly reduced the proportion of patients with low-grade cytological abnormalities, in comparison to placebo. Given the obtained level of significance (p < 0.001), the presented results indicate that short-term ALA supplementation shows a clinically significant effect on cervical cytology. Future studies should focus on the use of innovative formulations of ALA that might induce bioavailability and therapeutic efficiency against HPV infection and the investigation of synergistic effects of concurrent dietary/lifestyle modification and ALA supplementation in both low-grade and high-grade SIL.

Increasing use of nano-enabled products provides many benefits in various industrial processes and medical applications, but it also raises concern about release of nanoparticles (NPs) into the environment and subsequent human exposure. While potential toxicity of individual NPs types has been well described in scientific literature, exposure and health-related effects of nanomixtures has been poorly described. This study aimed to evaluate the combined effect of silver (AgNP) and polystyrene NPs (PSNP) on the human macrophages. AgNP are one of the most commercialized NPs due to efficient antimicrobial activity, while PSNP are ubiquitous in terrestrial and aquatic environments due to plastic pollution and degradation of polystyrene-based products. Differentiated monocytic cell line THP-1 were used as an in vitro model of human macrophages. Multiple aspects of cellular response to AgNP-PSNP nanomixture were analyzed including cell death, induction of apoptosis, oxidative stress response, expression of pro- and anti-inflammatory cytokines, and nanomechanical properties of cells. NPs uptake was visualized by confocal microscopy and quantified using flow cytometry. Results show that nanomixture increased apoptosis and cell death, expression of IL-6, IL-8 and TNFa, oxidative stress and mitochondrial dysfunction in cells compared to AgNP and PSNP applied as single treatments, indicating mixture additive action. Anti-inflammatory cytokines IL1b, IL-4 and IL-10 were not affected by combined exposure compared to single NPs. Visualization of NPs uptake and internalization showed that AgNP and PSNP were localized mostly in cytoplasm, with small fraction of AgNP translocated into cell nuclei, which explain increased number of double-stranded DNA breaks following exposure of cells to AgNPs alone or in the mixture. Study outcomes represent clear warnings on the human co-exposure to AgNP and PSNP that needs to be implemented in risk assessment approaches towards toxic-free environment.

Background: olive pomace extract (OPE) is a rich source of health promoting polyphenols (hydroxytyrosol (HTS) and tyrosol (TS)) and can be used as a nutraceutical ingredient of dietary supplements and functional foods. Its adequate bioavailability is a prerequisite for excreting biological activity and can be significantly and specifically affected by different food matrices. Methods: in order to investigate food effects on polyphenol bioaccessibility, OPE was co-digested with different foods according to internationally harmonized in vitro digestibility method. Impact of particular nutrients on HTS and TS permeability was assessed on Caco-2 cell monolayer. Results: HTS and TS bioaccessibility and transepithelial permeability can be significantly affected by foods (nutrients), especially by casein and certain types of dietary fiber. Those effects are polyphenol-and nutrient-specific and are achieved either through complexation in gastrointestinal lumen and/or through direct effects of nutrients on intestinal monolayer. Conclusions: obtained results emphasize the significance and complexity of polyphenol interactions within the food matrix and the necessity of individual investigational approaches with respect to particular food/nutrient and interacting phenolic compounds.

Aim To evaluate the impact of pharmacotherapeutic education on 30-day post-discharge medication adherence and adverse outcomes in patients with type 2 diabetes mellitus (T2DM). Methods The prospective, randomized, single-center study was conducted at the Medical Department of University Hospital Dubrava, Zagreb, between April and June 2018. One hundred and thirty adult patients with T2DM who were discharged to the community were randomly assigned to either the intervention or the control group. Both groups during the hospital stay received the usual diabetes education. The intervention group received additional individual pre-discharge pharmacotherapeutic education about the discharge prescriptions. Medication adherence and occurrence of adverse outcomes (adverse drug reactions, readmission, emergency department visits, and death) were assessed at the follow-up visit, 30 days after discharge. Results The number of adherent patients was significantly higher in the intervention group (57/64 [89.9%] vs 41/61 [67.2%]; χ2 test, P = 0.003]. There was no significant difference between the groups in the number of patients who experienced adverse outcomes (31/64 [48.4%] vs 36/61 [59.0%]; χ2 test, P = 0.236). However, higher frequencies of all adverse outcomes were consistently observed in the control group. Conclusion Pharmacotherapeutic education of patients with T2DM can significantly improve 30-day post-discharge medication adherence, without a significant reduction in adverse clinical outcomes. ClinicalTrial.gov identification number: NCT03438162

Background Despite numerous investigations, we still do not have a specific marker for pancreatic ductal adenocarcinoma. Only guideline-recommended biomarker for pancreatic ductal adenocarcinoma is the CA19-9, but it is also present in other gastrointestinal diseases. IMP3 is a new potential biomarker that is over-expressed in some cancers. The aims of our study were (1) to assess IMP3 in benign pancreatic lesions and pancreatic cancer, and (2) to estimate its concentrations in localized and advanced pancreatic cancer. Patients and methods Seventy-five patients with solid pancreatic lesions who underwent EUS-FNA were included. Patients were divided into three groups: benign lesions, cancer localized only on the pancreas, and patients with advanced pancreatic cancer (locally advanced or with distal metastases). Immunoreactivity of IMP3 was assessed on cytological smears sampled by endoscopic ultrasound. Results IMP3 was expressed in 89% of the patients with pancreatic cancer and not in benign lesions. Stronger expression of IMP3 protein and stage of the pancreatic cancer was statistically significant. IMP3 was expressed in all localized cancers and in 85% of patients with advanced pancreatic cancer. In the subgroup with locally advanced cancer, IMP3 was expressed in 88%, and in 83% of patients in the subgroup with distal metastasis (P = 0.007). In the present study, sensitivity was 89%, specificity 100%, with positive predictive value of 100% and negative predictive value of 63%. Conclusion There is a positive correlation between IMP3 expression and TNM stages of the pancreatic cancer. Higher expression of IMP3 on EUS-FNA specimens can suggest poorer prognosis.

Cytochrome P450 3A4 is the most significant enzyme in metabolism of medications. Flavonoids are common secondary plant metabolites found in fruits and vegetables. Some flavonoids can interact with other drugs by inhibiting cytochrome P450 enzymes. Thus, the objective of this study was to determine inhibition kinetics of cytochrome P450 3A4 by flavonoids: acacetin, apigenin, chrysin and pinocembrin. For this purpose, testosterone was used as marker substrate, and generation of the 6β-hydroxy metabolite was monitored by high performance liquid chromatography coupled with diode array detector. IC50 values, inhibition constants, and rates of inhibition were determined. IC50 values ranged between 0.6 and 11.4 µM. The strongest inhibitor was chrysin (IC50 0.6 µM, inhibition constant 0.6 µM, inhibition rate constant 0.065 min–1, inhibition efficacy 0.108 min–1 µM–1). Compared to other flavonoids analyzed, chrysin’s inhibitory effect can be attributed to the hydrophobic nonsubstituted B ring, as well as rigidity of the structure. When foods rich in chrysin are consumed, e.g. honey and propolis, chrysin can cause food-drug interactions. Further in vitro studies are needed to determine the reactive intermediate responsible for inactivation of cytochrome P450 3A4 enzyme, as well as in vivo studies to determine possible clinical significance of this inhibition. This work is licensed under a Creative Commons Attribution 4.0 International License.

An adequate level of low molecular weight thiols (LMW-SH, especially glutathione (GSH)) protects cellular macromolecules against toxic agents, and is used as a sensitive biomarker of exposure to toxic compounds. During sample collection, storage and preparation, non-enzymatic and enzymatic oxidation of LMW-SH can occur leading to analytical inaccuracy. The aim of this study was to optimize a fast and reliable screening method for the determination of LMW-SH, mainly GSH, in blood and plasma samples as well as to investigate the impact of storage conditions on the LMW-SH stability. Based on our results, the described spectrophotometric method allows fast and reliable determination of LMW-SH in blood and plasma samples. Results on incubation of samples at 37 °C imply that synthesis of LMW-SH (probably GSH) as well as dynamic interexchange among various thiols forms can be induced in blood cells in in vitro conditions. Importantly, the level of LMW-SH in blood and plasma stored at −20 °C was constant, indicating that they can be stored at −20 °C for at least 30 days. Therefore, the method is suitable for assessment of LMW-SH in long-term human biomonitoring as well as environmental field studies, especially those involving a large number of samples such as epidemiological studies.

The purpose of this paper was to outline the development of short peptide targeting of the human prostate specific antigen (hPSA), and to evaluate its effectiveness in staining PSA in human prostate cancer tissue. The targeting of the hPSA antigen by means of antisense peptide AVRDKVG was designed according to a three-step method involving: 1. The selection of the molecular target (hPSA epitope), 2. the modeling of an antisense peptide (paratope) based on the epitope sequence, and 3. the spectroscopic evaluation of sense&ndash, antisense peptide binding. We then modified standard hPSA immunohistochemical staining practice by using a biotinylated antisense peptide instead of the standard monoclonal antibody and compared the results of both procedures. Immunochemical testing on human tissue showed the applicability of the antisense peptide technology to human molecular targets. This methodology represents a new approach to deriving peptide ligands and potential lead compounds for the development of novel diagnostic substances, biopharmaceuticals and vaccines.

25-Hydroxyvitamin D (25-OHD) may have a prognostic value in colorectal cancer (CRC) patients. However, as 25-OHD concentration is strongly impacted by surgery, it is uncertain what is the most reliable time-point for 25-OHD assessment, pre- or post-operative. Therefore, we examined 515 CRC patients (AJCC I-III) who underwent surgery. Blood samples were collected either pre- operatively (n = 286 ; median = 1 day before surgery) or post-operatively (n = 229 ; median = 8 days). Serum 25-OHD concentration was determined by liquid chromatography-tandem mass spectrometry. Association between 25-OHD and survival was tested in the whole cohort, followed by stratified analyses in pre- and post-operatively sampled. Median 25-OHD in the cohort was 36.7 nmol/L and median follow-up time was 5.9 years. There were no differences between pre- and post-operative cohort in age, sex, 25- OHD, AJCC stage, or localization of tumor. After adjustment, higher 25-OHD (>50 nmol/L) was associated with better overall survival only in post-operative (HR = 0.53 ; 95% CI: 0.33-0.84 ; P = 0.006), but not in pre- operative cohort (HR = 1.13 ; 95% CI: 0.77-1.65 ; P = 0.53). In conclusion, higher post-operative 25-OHD levels were associated with better survival outcome in CRC patients, while no such association was found for pre-operative levels. Time-point of blood collection should be addressed carefully in future research as it might affect the prognostic value of 25-OHD in CRC.

Many personal care products on the market contain endocrine disrupting chemicals, including parabens. Parabens are well known chemical additives used as preservatives. They have been found in mammary glands and breast cancer tissues. At the same time, the general public is increasingly exposed to plastic micro- and nanoparticles generated during plastic production and waste disposal. Exposure to chemical cocktails is a realistic scenario of high public health interest, in which many types of compounds such as these two may exhibit synergistic or additive adverse effects. This study evaluated the effects of plastic nanoparticles, parabens, and their mixture on the viability and proliferation of two human breast cancer cell lines: MDA-MB 231, which lacks oestrogen receptors, and MCF-7, which expresses these receptors. Parabens increased proliferation of oestrogen-sensitive breast cancer cells, and this effect became synergistic in the presence of plastic nanoparticles. The mechanism behind synergy may be related to the translocation and adsorption properties of nanoplastics, which served as a Trojan horse to expose cells to parabens more efficiently. These preliminary findings support growing evidence warning about the urgent problem of human exposure to combinations of plastic waste and contingent chemicals., Većina proizvoda za osobnu njegu na tržištu sadrži kemikalije koje remete rad endokrinoga sustava kao što su parabeni. Primjena takvih proizvoda povećava izloženost opće populacije takvim kemikalijama. Parabeni, dobro poznati aditivi koji se koriste kao konzervansi, otkriveni su u mliječnim žlijezdama i tkivima raka dojke. Osim takvih kemikalija, izloženost plastičnim mikro- i nanočesticama koje nastaju tijekom proizvodnje, uporabe i odlaganja plastičnog otpada velik su zdravstveni i okolišni problem koji treba hitno rješavati. Za pouzdanu procjenu rizika za ljudsko zdravlje, nužno je objasniti ne samo učinke pojedinačnih kemijskih spojeva nego i njihovih mješavina. Istraživanje izloženosti kemijskim koktelima može pružiti podatke o mogućem sinergističkom ili aditivnom učinku kemikalija u takvim koktelima. Cilj ovoga istraživanja bio je procijeniti učinak plastičnih nanočestica, parabena i njihovih koktela na vijabilnost i proliferaciju dviju različitih stanica karcinoma dojke: stanične linije MDA-MB 231 kojoj nedostaju estrogeni receptori i MCF-7 stanica koje izražavaju te receptore. Dobiveni rezultati pokazali su da ni parabeni ni plastične nanočestice, primijenjeni posebno, u ispitivanim koncentracijskim rasponima nisu značajnije promijenili vijabilnost i proliferacijsku aktivnost u stanicama karcinoma dojke. Međutim, parabeni su pojačali proliferacijsku aktivnost stanica osjetljivih na estrogene koja je sinergistički potaknuta prisutnošću plastičnih nanočestica. Mehanizam takva djelovanja može biti povezan s translokacijskim i adsorpcijskim svojstvima nanočestica, koje su poslužile kao vektor za učinkovitije izlaganje stanica parabenima. Protumorska aktivnost na karcinomu dojke može se očekivati kao konačni ishod izloženosti takvom kemijskom koktelu. Ovi preliminarni podatci dragocjeni su dodatak postojećim ranim upozorenjima na novi problem s povećanjem izloženosti ljudi plastičnom otpadu i kemijskim kemikalijama.

Psoriasis is a common chronic inflammatory skin disease which affects 0.5–1 % of children and 2–3 % of the adult population. In Croatia, 1.6 % of the population suffer from psoriasis. Distribution of the disease is bimodal, with the first peak at the age of 20–30, and the second at the age of 50–60. The etiopathogenesis of the disease is multifactorial, the key factors being genetic predisposition combined with immunological disorders, environmental factors and skin barrier damage. There are several clinical variants of the disease. The main signalling pathways in psoriasis include TNF-α, IL-23 and IL-17. Topical agents are used for the treatment of the mild form, and the systemic conventional therapy is used for the treatment of moderate to severe forms of the disease. In cases where’s no response, or intolerance or contraindications are present, new targeted medications are to be administered. Development in the field of immunogenetics of psoriasis leads to personalized medicine.

Metallothioneins are peculiar cysteine rich, heat resistant, small cellular plasma proteins expressed through almost all life forms. The currently established biological functions of metallothioneins are the homeostasis of essential metals and protection against toxic transitional metals (TM) alongside defence from oxidative stress by direct scavenging of reactive oxygen and nitrogen species (ROS and RNS). In mammals, among the four main evolutionary conserved forms, only the ubiquitously expressed metallothionein 1 and 2 (here abbreviated as MT) are inducible by TM, oxidative stress, glucocorticoids and starvation among various other stimuli. However, more than sixty years after being discovered, metallothioneins still bear unresolved issues about their possible physiological function and regulation. The biological function of MTs has still not been associated with the in vitro-demonstrated capacity of MT interaction with cellular molecules glutathione (GSH) or adenosine triphosphate (ATP), or with the possibility of direct iron-MT binding in the reducing intracellular environment of some organelles, e.g. lysosomes. Iron as the most abundant cellular TM is also one of the main physiological sources of ROS. Moreover, iron exhibits strain, sex and age differences that reflected ROS generation and MT induction in (patho)physiology and toxicology studies. A recent study showed that iron sex differences follows expression of both ferritin and MT leading to wide implications from essential TM interconnectivity to aging. This review places emphasis on biochemically proven but physiologically ignored interactions of MT with iron to stimulate advanced research for establishing a wide frame of the biological roles of MTs important for health and longevity., Metalotioneini su mali proteini stanične plazme specifični po zastupljenosti cisteina i otpornosti na toplinu, a izraženi su kroz gotovo sve oblike života. U sisavaca, među četirima glavnim evolucijski očuvanim oblicima, samo se sveprisutno izraženi metalotioneini 1 i 2 (u daljnjem tekstu MT) induciraju s različitim prijelaznim metalima (TM), oksidacijskim stresom, glukokortikoidima, gladovanjem između ostalih stimulansa. Trenutna uspostavljena uloga MT-ova u stanici je homeostaza esencijalnih i zaštita od toksičnih TM-ova, zajedno s obranom od oksidacijskoga stresa izravnim uklanjanjem reaktivnih kisikovih i dušikovih vrsta (ROS/RNS). Međutim, čak i više od šezdeset godina nakon otkrića, oko metalotioneina i dalje postoje neka neriješena pitanja o njihovoj funkciji i regulaciji. Naime, funkcija MT-ova još uvijek nije povezana s in vitro dokazanom sposobnošću vezanja MT-ova sa staničnim molekulama GSH-a ili ATP-a, zajedno s važnim procesom vezanja željeza MT-a u reduktivnom okolišu npr. lizosoma. Željezo kao najzastupljeniji stanični TM također je i glavni izvor ROS-a. Ujedno željezo pokazuje razlike između vrsta i sojeva te među spolovima i u starenju, koje mogu odražavati nastajanje ROS-ova i izražaj MT-ova u (pato)fiziološkim i toksikološkim istraživanjima. Promjene MT-ova koje prate razlike u željezu nedavno su dokazane i imaju široke implikacije od međusobne povezanosti esencijalnih TM-ova do starenja. Ovaj pregledni članak stavlja naglasak na biokemijski dokazane ali većinom fiziološki ignorirane interakcije in vitro MT-ova i željeza za poticaj naprednih istraživanja koja bi izgradila širu mrežu MT-ove biološke uloge važne za zdravlje i dugovječnost.

Melanocortins, i.e., melanocyte stimulating hormones (MSH) are peptides with strong antiinflammatory effects. The most investigated aspects of γ2-MSH are related to cardiovascular effects and natriuresis, with limited research available about its anti-inflammatory and cytoprotective effects. The aims of this study were: 1) to examine the effects of γ2-MSH and its derivative [D-Trp(8)]-γ2-MSH on the acetaminophen model of liver damage in CBA mice; 2) to evaluate the modulation of γ2-MSH hepatoprotection by melanocortin subtypes 3 and 4 receptor antagonists SHU 9119 and HS 024; 3) to define the importance of central MSH pharmacophore region (HFRW) by using antisense peptides LVKAT and VKAT. In this study, specific antagonists and antisense peptides were used to target central pharmacophore region of γ2-MSH and [D-Trp(8)]-γ2-MSH, enabling the evaluation of hepatoprotection from the standpoint of the receptor and pharmacophore blockade. The criteria for monitoring the effects of the hormones on the liver damage were alanine transaminase, aspartate transaminase activities (U/L), and pathohistological scoring of liver necrosis (scale 0-5). γ2-MSH (0.24 mg/kg) indicated hepatoprotective effects in comparison to control (p < 0.001). In contrast, [D-Trp(8)]-γ2-MSH did not show any hepatoprotective effects. Application of antagonists SHU 9119 and HS 024, and antisense peptides LVKAT and VKAT, also did not show any hepatoprotective effects. In fact, when combined with γ2-MSH, it annulled its hepatoprotective effect. The results provide evidence for hepatoprotective and antiinflammatory effects of the γ2-MSH in the liver.

Sense and antisense peptides, i.e. peptides specified by complementary DNA and RNA sequences, interact with increased probability. Biro, Blalock, Mekler, Root-Bernstein and Siemion investigated the recognition rules of peptide—peptide interaction based on the complementary coding of DNA and RNA sequences in 3′ → 5′ and 5′ → 3′ directions. After more than three decades of theoretical and experimental investigations, the efficiency of this approach to predict peptide—peptide binding has been experimentally verified for more than 50 ligand—receptor systems, and represents a promising field of research. The natural genetic coding algorithm for sense and antisense peptide interactions combines following elements: of amino acid physico- chemical properties, stereochemical interaction, and bidirectional transcription. The interplay of these factors influences the specificity of sense—antisense peptide interactions, and affects the selection and evolution of peptide ligand—receptor systems. Complementary mRNA codon—tRNA anticodon complexes, and recently discovered Carter- Wolfenden tRNA acceptor-stem code, provide the basis for the rational modeling of peptide interactions based on their hydrophobic and lipophilic amino acid physico-chemical properties. It is shown that the interactions of complementary amino acid pairs according to the hydrophobic and lipophilic properties strongly depend on the central (second) purine base of the mRNA codon and its pyrimidine complement of the tRNA anticodon. This enables the development of new algorithms for the analysis of structure, function and evolution of protein and nucleotide sequences that take into account the residue's tendency to leave water and enter a nonpolar condensed phase considering its mass, size and accessible surface area. The practical applications of the sense—antisense peptide modeling are illustrated using different interaction assay types based on: microscale thermophoresis (MST), tryptophan fluorescence spectroscopy (TFS), nuclear magnetic resonance spectroscopy (NMR), and magnetic particles enzyme immunoassay (MPEIA). Various binding events and circumstances were considered, e.g., in situations with—short antisense peptide ligand (MST), L- and D-enantiomer acceptors (TFS), in low affinity conditions (NMR), and with more than one antisense peptide targeting hormone (MPEIA).

Antisense peptide technology is a valuable tool for deriving new biologically active molecules and performing peptide–receptor modulation. It is based on the fact that peptides specified by the complementary (antisense) nucleotide sequences often bind to each other with a higher specificity and efficacy. We tested the validity of this concept on the example of human erythropoietin, a well-characterized and pharmacologically relevant hematopoietic growth factor. The purpose of the work was to present and test simple and efficient three-step procedure for the design of an antisense peptide targeting receptor-binding site of human erythropoietin. Firstly, we selected the carboxyl-terminal receptor binding region of the molecule (epitope) as a template for the antisense peptide modeling, Secondly, we designed an antisense peptide using mRNA transcription of the epitope sequence in the 3'→5' direction and computational screening of potential paratope structures with BLAST, Thirdly, we evaluated sense–antisense (epitope–paratope) peptide binding and affinity by means of fluorescence spectroscopy and microscale thermophoresis. Both methods showed similar Kd values of 850 and 816 µM, respectively. The advantages of the methods were: fast screening with a small quantity of the sample needed, and measurements done within the range of physicochemical parameters resembling physiological conditions. Antisense peptides targeting specific erythropoietin region(s) could be used for the development of new immunochemical methods. Selected antisense peptides with optimal affinity are potential lead compounds for the development of novel diagnostic substances, biopharmaceuticals and vaccines.

Nealkoholna masna bolest jetre (NAFLD; engl. non-alcoholic fatty liver disease) predstavlja jednu od najčešćih kroničnih upalnih bolesti jetre koja može prijeći u steatohepatitis, koji u oko 20 % slučajeva progredira do ciroze ili karcinoma jetre. Radi se o multifaktorijalnoj bolesti za koju trenutno nema zadovoljavajuće terapije, a farmakološke metode se baziraju uglavnom na liječenju metaboličkih poremećaja koji su povezani s bolešću. Koriste se antidijabetici koji povećavaju osjetljivost na inzulin (tiazolidini i bigvanidi), antioksidansi (vitamin E), antilipemici (statini), blokatori angiotenzinskih receptora i drugi, međutim niti jedan od njih nije pokazao značajnu učinkovitost i dugotrajnu sigurnost u primjeni. Zbog svega navedenog, posljednjeg desetljeća, biljni preparati sve više privlače pažnju zbog široke dostupnosti, relativno malo nuspojava, širokog mehanizma djelovanja, te posljedične koristi. Dosad objavljene in vitro i in vivo studije pokazuju da biljni preparati kroz različite mehanizme djelovanja smanjuju oštećenje hepatocita povezano s NAFLD. Neki od navedenih mehanizama su: smanjenje lipogeneze; povećanje oksidacije β-masnih kiselina; povećanje osjetljivosti na inzulin; smanjenje oksidativnog stresa i inhibicija aktivacije upalnih putova. Biljni preparati se mogu koristiti kao biljni ekstrakti (korijen sladića, zeleni čaj, sjeme sikavice, crveno grožđe),kemijske tvari (flavonoidi, polifenoli, saponini, alkaloidi) i biljne mješavine (kineska, japanska). Iz invitro studija i in vivo životinjskih modela, možemo zaključiti da biljni lijekovi mogu biti obećavajuća terapijska sredstva, no vrlo je malo randomiziranih kliničkih studija koje bi potvrdile njihovu učinkovitost i sigurnost u ljudi.Veliki problem predstavlja i standardizacija uzorka, formulacija te doziranje., Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic inflammatory diseases of the liver, which can be transmitted to steatohepatitis, which in about 20% progresses to cirrhosis or liver cancer. It is a multifactorial disease for which there is currently no satisfactory therapy, and pharmacological methods are based mainly on the treatment of metabolic disorders associated with the disease. Antidiabetic agents that increase insulin sensitivity (thiazolidines and biguanides), antioxidants (vitamin E), antiliphemics (statins), angiotensin receptor blockers and others are used, however none of them showed significant efficacy and long-term safety in use. Because of all this, for the last decade, herbal preparations are increasingly attracting attention due to their wide availability, relatively few side effects, wide mechanism of action, and consequent benefits. So far published in vitro and in vivo 2 Sažeci radova / Abstracts studies have shown that herbal preparations through various mechanisms of action reduce hepatocellular damage associated with NAFLD. Some of the mechanisms mentioned are: reduction of lipogenesis; increased oxidation of β-fatty acids; insulin sensitivity increase; reduction of oxidative stress and activation of inflammatory pathways. Herbal preparations can be used as herbal extracts (herb root, green tea, glitter seeds, red grapes), chemical substances (flavonoids, polyphenols, saponins, alkaloids) and herbal mixtures (chinese, japanese). From in vitro studies and in vivo animal models, we can conclude that herbal remedies can be promising therapeutic agents, but there are very few randomized clinical studies that would confirm their effectiveness and safety in humans. A major problem is the standardization of the sample, formulation and dosage.

Recently discovered anti-inflammatory and immunomodulatory properties of melanocortin peptides led to the conclusion that they might serve as new anti-inflammatory therapeutics. The purpose of this work was to examine the effectiveness of β-melanocortin (β-MSH) in two experimental models: ethanol-induced gastric lesions and TNBS (2, 4, 6- trinitrobenzenesulfonic acid)-induced colitis in male Wistar rats. Three progressive doses of β-MSH were used: 0.125, 0.250 and 0.500 mg/kg. Our results suggest that β-MSH acts as a protective substance in the gastric lesions model, which can be seen as a statistically significant reduction of hemorrhagic lesions at all three doses, compared to the control group. The most efficient dose was 0.250 mg/kg. Statistically significant reduction in mucosal surface affected by necrosis and the reduction of overall degree of inflammation in the colitis model indicates an anti-inflammatory effect of β-MSH at a dose of 0.250 mg/kg. The results justify further research on β-MSH peptide and its derivates in the inflammatory gastrointestinal diseases, and point out the possibility of using β-MSH in studies of digestive system pharmacology.

Proteins and peptides in mammals are based exclusively on L-amino acids. Recent investigations show that D-amino acids exhibit physiological effects in vivo, despite of their very small quantities. We have investigated the hepatoprotective effects of the Land D-enantiomers of alpha-melanocortin peptide (alpha-MSH). The results showed that peptide-enantiomerism is related to the protective effects of melanocortin peptides in vivo. L-alpha-MSH exhibited potent hepatoprotective effect in the experimental model of acetaminophen induced hepatotoxicity in male CBA mice, while its D-mirror image was inefficient. Furthermore, the antibody to the L-peptide did not recognize the D-structure. The results indicate that the opposite peptide configuration may be used to modulate its function and metabolism in vivo and in vitro.

Recent histopathological investigations in patients with hepatitis suggested possible involvement of Met-enkephalin and its receptors in the pathophysiology of hepatitis. Consequently, we evaluated the potential hepatoprotective effects of this endogenous opioid pentapeptide in the experimental model of acetaminophen induced hepatotoxicity in male CBA mice. Met-enkephalin exhibited strong hepatoprotective effects in a dose of 7.5 mg/kg, which corresponds to the protective dose reported for several different animal disease models. In this group plasma alanine aminotransferase and aspartate aminotransferase enzyme activities, as well as liver necrosis score were significantly reduced in comparison to control animals treated with physiological saline (p &gt, 0.01). The specificity of the peptide hepatoprotection was investigated from the standpoint of the receptor and peptide blockade. It was concluded that Met-enkephalin effects on the liver were mediated via δ and ζ opioid receptors. Genotoxic testing of Met-enkephalin confirmed the safety of the peptide.

Background: Metal allergies can be clinical problem, especially in atopic individuals. This study is unique and contributes with new knowledge in everyday life skin care of irritant and atopic dermatitis patients. Objectives: The aim of the study was to determine the frequency of positive patch test reactions to metal contact allergens (potassium dichromate, cobalt chloride, nickel sulphate, white mercury precipitate) in patients diagnosed with allergic contact dermatitis, irritant contact dermatitis, and atopic dermatitis. Patients and Methods: Between 2007 and 2011, patch testing was performed in 2185 patients according to International Contact Dermatitis Research Group technique. Results: Study results showed statistically significant differences in patch test responses to 2 allergens, nickel sulphate (χ2 = 24.22; p < 0.001) and cobalt chloride (χ2 = 22.72; p < 0.001). Nickel sulphate was the most common allergen in allergic contact dermatitis and atopic dermatitis, while for irritant contact dermatitis the most common allergen was cobalt chloride. Among the 4 tested metal allergens the most common and relevant was nickel sulphate (χ2 = 17.25; p < 0.004), found in almost all examinees. Conclusion: The increased awareness of allergens and their potential sources may help to limit the use of these chemicals in consumer product manufacturing.

Background: L-alpha-Melanocortin is a strong inhibitor of inflammation.It is a promising new anti-inflammatory and hepatoprotective peptide. Consequently, its melanocortin receptors (MC1, MC3, MC4 and MC5) could be possible targets for the development of new antiinflammatory drugs for chronic inflammatory liver disease. For a long ime it has been believed that only the L-enantiomers of amino acids are present in higher animals, but recent investigations show that D-amino acids also exhibit physiological effects in vivo, despite their very small quantities. The aim of this study was to compare hepatoprotective effects of L-alphamelanocortin and D-alpha-melanocortin using the acetaminophen model of chemical liver damage in male CBA mice. Methods: Tested substances were applied intraperitoneally 60 minutes prior to the intragastric application of acetaminophen (150 mg/kg). Animals were sacrificed 24 hours after the administration of acetaminophen. The criteria for monitoring hepatoprotective effects of the tested substances were biochemical parameters (AST and ALT) and histopathological analysis. Results: The results obtained by the histopathological analysis and biochemical findings show potent hepatoprotective and anti-inflammatory effects of L-alpha-melanocortin in the liver, and suggest the possibility of modulating liver inflammation by means of melanocortin molecules and related receptors. D-alpha-melanocortin did not show any hepatoprotective effects in vivo. Conclusions: Our results show that peptide enantiomerism influences the protective effects of alpha-melanocortin peptides in vivo. This concept may be used to modulate peptide function in vivo and antibody binding assay in vitro. Acknowledgements: The support of the Croatian Ministry of Science, Education and Sports is gratefully acknowledged (grant no. 098-0982929-2524).

The genetic code defines nucleotide patterns that code for individual amino acids and their complementary, i.e., antisense, pairs. Peptides specified by the complementary mRNAs often bind to each other with a higher specificity and efficacy. Applications of this genetic code property in biomedicine are related to the modulation of peptide and hormone biological function, selective immunomodulation, modeling of discontinuous and linear epitopes, modeling of mimotopes, paratopes and antibody mimetics, peptide vaccine development, peptidomimetic and drug design. We have investigated sense-antisense peptide interactions and related modulation of the peptide function by modulating the effects of a-MSH on hepatoprotection with its antisense peptide LVKAT. First, transcription of complementary mRNA sequence of a-MSH in 3’→5’ direction was used to design antisense peptide to the central motif that serves as a-MSH pharmacophore for melanocortin receptors. Second, tryptophan spectrofluorometric titration was applied to evaluate the binding of a-MSH and its central pharmacophore motif to the antisense peptide, and it was concluded that this procedure represents a simple and efficient method to evaluate sense-antisense peptide interaction in vitro. Third, we showed that antisense peptide LVKAT abolished potent hepatoprotective effects of a-MSH in vivo.

Cilj: Cilj istraživanja bio je usporediti rezultate toalete nosnih šupljina proizvodom za nazalnu higijenu Sterimar u odnosu na nosnu toaletu fiziološkom otopinom ili izostanak uporabe ikakve lokalne toalete u bolesnika nakon operacije tumora selarne regije. Bolesnici i metode: Na temelju subjektivne ocjene bolesnika provedeno je istraživanje osam parametara na 90 bolesnika nakon transnazalne transsfenoidalne operacije hipofize u trajanju od 30 dana. Bolesnici su bili podijeljeni u 3 skupine: skupina koja je toaletu nosa provodila fiziološkom otopinom, skupina koja je upotrebljavala izotoniènu morsku vodu (Sterimar sprej) i treæa skupina bolesnika koji nisu provodili nikakvu toaletu nosnica. Rezultati: U periodu izmeðu 10. i 15. dana nakon detamponade u bolesnika koji su toaletu nosa provodili sprejem Sterimar zapaženo je statistièki znaèajno smanjena kolièina prednjeg i stražnjeg sluzavog iscjetka iz nosa, što je olakšalo disanje na nos i popravilo kvalitetu života. Rasprava i zakljuèak: Kod najveæeg broja bolesnika došlo je do poboljšanja analiziranog svojstva tijekom razdoblja ispitivanja u trajanju od 30 dana. To poboljšanje osobito se manifestiralo u skupini bolesnika koji su toaletu nosnih šupljina obavljali Sterimarom sa statistièki znaèajnim smanjenjem prednjeg i stražnjeg iscjetka iz nosa., Aim: In this research we are comparing the results of nasal packing with Sterimar solution to the results obtained with saline lavage or no local packing after transsphenoidal pituitary surgery. Patients and methods: This investigation was based on subjective assessment among 90 patients operated for transnasal transsphenoidal pituitary surgery. The patients were divided into 3 groups according to type of postoperative nasal care and follow up was 30 days. The first group used saline solution, the second group used isotonic sea water (Sterimar spray) and the third group did not use and nasal irrigation postoperatively. Results: Statistically significant improvement in decreased anterior and posterior nasal secretion was observed in Sterimar group of patients between day 10. And 15. Resulting in improved nasal breathing and quality of life in that period. Discussion and conclusion: Most patients had an improvement of the monitored feature during the period od 30 days. This improvement was most pronounced in the group of patients which have used Sterimar solution who had a statistically significant decrease of anterior and posterior nasal secretion.ijih st

Hitna stanja u alergologiji i kliničkoj imunologiji sve su češći medicinski javno zdravstveni problem u nas i u svijetu. Česte su alergijske reakcije od blagih simptoma kao što je svrbež do generaliziranih kožnih, gastrointestinalnih, respiratornih simptoma do anafilaksije i zatajenja kardiovaskularnog sustava. Pri tom su moguće reakcije na različite udisajne tvari, hranu i aditive u hrani, strane tvari (krv, lateks i drugo) te brojne poznate antigene i lijekove koji putem imunskih kompleksa i reakcija posredovanih oslobađanjem komplementa ili direktnim oslobađanjem histamina (opijati ili dekstran) te zbog modulatora metabolizma arahidonske kiseline (npr. NSAID) i drugo. NSAID često uzrokuju astmu, urtikarijski osip, angioedem i još 35 različitih dermatoloških reakcija, posebice kod primjene lijekova koji zahtijevaju hitan terapijski postupak. Reakcije od uboda insekata pojavljuju se ne samo s bolnim infiltratima, nego i diseminiranim osipima i edemom. Suvremene dijagnostičke metode, a posebice hitni terapijski postupci, sukladno internacionalnom k oncenzusu (npr. European Allergy White Paper) ukazuju na potrebu ujednačavanja stavova u kliničkoj praksi. Velik broj bolesnika se liječi zbog alergijskih kožnih bolesti neadekvatno, upućuje na testiranja često u fazi aktivne bolesti i kad su pod terapijom, a liječe se kortikosteroidima i antihistaminicima duže no što je potrebno. Suvremeno liječenje hitnih stanja zahtijeva izobraženu osobu, no svaki liječnik treba odgovarati za liječenje akutnih simptoma u svojoj ordinaciji i poznavati postupak za hitno liječenje točno određenim lijekovima. Anafilaktične reakcije su rijetke, ali svaki liječnik treba biti pripravan na najteže situacije i poznavati liječenje u hitnom postupku. U radu navodimo najčešće bolesti za koje je potreban hitan postupak liječenja, a u fazi regresije bolesti dijagnostički postupci., Emergencies in allergology and clinical immunology are quite common and pose a public health problem in Croatia and worldwide. Allergic reactions ranging from mild symptoms such as pruritus to generalized skin eruptions, gastrointestinal and respiratory symptoms to anaphylaxis with cardiovascular emergencies are very common. Reaction develops to inhalant substances, food and food ingredients, various foreign substances (blood, latex, etc.) and many other well known antigens and drugs which are frequent causes of anaphylactic reactions, asthma, generalized urticaria, angioedema. There are 35 different skin reactions induced by drugs through immune complexes, complement mediated reactions and direct histamine liberation (by opiates or dextran) and modulators of arachidonic acid metabolism (such as NSAID). Along with painful infiltrates, insect venom allergy manifests with disseminated exanthema and angioedema. Current diagnostic methods, especially therapeutic procedures for emergencies in concordance with the international consensus of the European Allergy White Paper, require the same procedure in clinical practice. Many patients with different allergic skin diseases are treated inappropriately, some are tested in active phase of the disease, during anti-allergic therapy, and some are administered corticosteroids and antihistamines longer than necessary. Sometimes, the treatment of emergencies requires a person experienced in intensive care medicine. However, any physician might be responsible for the treatment of acute symptoms in his office, and thus has to know the modalities and relevant drugs and treatment essentials. Anaphylactic reactions are rare, but every physician should be prepared for the worst. Due knowledge of the correct management should be a must for all of us. The most common emergencies require emergency treatment and diagnostic procedures upon disease regression.

Research over the past decade has indicated that melanocortin peptides are potent inhibitors of inflammation and a promising source of new anti-inflammatory and cytoprotective therapies. The purpose of the present paper is to compare protective effects of alpha-, beta-, and gamma-melanocyte stimulating hormone on acetaminophen induced liver lesions in male CBA mice. Acetaminophen was applied intragastrically in a dose of 150 mg/kg, and tested substances were applied intraperitoneally 1 hour before acetaminophen. Mice were sacrificed after 24 hours and intensity of liver injury was estimated by measurement of plasma transaminase activity (AST and ALT) and histopathological grading of lesions. It was found that alpha-, beta-, and gamma-MSH decrease intensity of lesions by both criteria in a dose-dependent manner.

Cilj ovog prospektivnog ispitivanja bio je ispitati djelotvornost profilaktičke primjene probiotika Acidosalusa® u žena s rekurentnim cistitisom. Acidosalus® je jedinstvena kombinacija Lactobacillus acidophilus, Lactobacillus rhamnosus i bifidobakterija u visokim terapijskim koncentracijama živih bakterija spremnih za razmnožavanje. Sadrži nadalje vitamine B skupine, folnu kiselinu, lako topive laktate, kalij, željezo, fosfor, te ima mogućnost stvaranja antimikrobnih supstanci - mliječne i octene kiseline, diacetila, laktocidina, acidofilucina, acidocina i acidofilina. Ispitivanje je provedeno u Ambulanti za urogenitalne infekcije Klinike za infektivne bolesti „Dr. Fran Mihaljević“ u Zagrebu u razdoblju od 01.01.2006. do 31.12.2009. godine. Ispitivano je 117 bolesnica u dobi od 18 do 65 godina s dijagnozom rekurentnog cistitisa - s više od dvije epizode cistitisa u posljednjih 6, odnosno više od 3 epizode u posljednjih 12 mjeseci. Uvjeti za uključivanje su bili: prisutnost simptoma infekcije donjeg urotrakta (dizurija, polakizurija, urgencija), identičan mikrobiološki nalaz obriska rodnice i urinokulture, leukociturija, te ultrazvukom isključena abnormalnost mokraćnog sustava. Sve su bolesnice kroz 7 dana liječene adekvatnom antimikrobnom terapijom, a zatim su nasumce podijeljene u dvije skupine. U jednih je odmah po prestanku antimikrobne terapije primijenjena profilaksa s probiotikom Acidosalus® te su kroz najmanje tri mjeseca kontinuirano uzimale peroralno Acidosalus® uz istovremenu vaginalnu primjenu Acidosalus® vaginaleta kroz najmanje 7 dana. Planirana primjena profilakse bila je 3 mjeseca. Na kontroli 1-2 tjedna nakon završene sedmodnevne antimikrobne terapije, sve su bolesnice bile klinički i bakteriološki izliječene. Do kontrole 3 mjeseca nakon završene sedmodnevne antimikrobne terapije u skupini od ukupno 56 žena koje su primale profilaksu probiotikom rekurentne epizode cistitisa imalo je njih 3 (5,4%), a u skupini od ukupno 61 žene koje nisu primale profilaksu broj rekurentnih epizoda bio je 11 (18,3%). Tri mjeseca nakon završene 7-dnevne antimikrobne terapije uropatogene bakterije su dokazane češće u rodnici žena koje nisu primale profilaksu probiotikom (u 44 od ukupno praćenih 50 bolesnica - 88%) nego u žena koje su primale profilaksu probiotikom (u 18 od ukupno 53 bolesnica - 34%). Rezultati ove studije pokazuju da je profilaktička primjena mješovite kulture probiotika sadržane u medicinskom proizvodu Acidosalus® u žena s rekurentnim uroinfekcijama djelotvorna i sigurna. Ističemo važnost istovremene oralne i vaginalne primjene, čime se uspostavlja obnavljanje i poboljšanje funkcije autohtone mikroflore crijeva i rodnice, smanjuje rezervoar uropatogenih bakterija u crijevu i jača imunitet općenito., The aim of this prospective research was to investigate the efficacy of prophylactic use of probiotic Acidosalus® in women with recurrent cystitis. Acidosalus® is a unique combination of Lactobacillus acidophilus, Lactobacillus rhamnosus and bifidobacteria in high therapeutic concentrations of live bacteria ready for multiplication. It consists of group B vitamins, folic acid, highly soluble lactates, potassium, iron, phosphorus, and has the possibility to produce antimicrobial substances - lactic and acetic acid, diacetyl, lactocidine, acidophilucin, acidocin and acidophylin. The research was conducted at the Outpatient Department for Urogenital Infections of the University Hospital for Infectious Diseases „Dr. Fran Mihaljević“ in Zagreb in the period between 01.01.2006. until 31.12.2009. We investigated 117 patients aged between 18 and 65 years with diagnosed recurrent cystitis – more than two episodes of cystitis in the past 6, i.e. more than three episodes in the past 12 months. The inclusion criteria were as follows: presence of symptoms of lower urinary tract infection (dysuria, polakisuria, urgency), identical microbiological finding of cervical swab and urinary culture, leukocyturia, ultrasound excluded urinary tract abnormality. All patients received adequate antimicrobial therapy for 7 days and then were randomly divided into two groups. One group received Acidosalus® probiotic as a prophylaxis immediately after the end of antimicrobial therapy – and at least three months continuously received Acidosalus® orally with simultaneous vaginal administration of Acidosalus® vaginalettes for at least 7 days. Planned duration of prophylaxis was three months. Follow up visit performed 1-2 weeks after completion of seven day course of antimicrobial therapy showed that all patients were clinically and bacteriologically cured. Until the follow up visit three months after completed 7-day antimicrobial therapy, recurrent episodes of cystitis were recorded in 3 (5,4%) out of a total of 56 women who received probiotic prophylaxis and in 11 (18,3%) out of a total of 61 women who did not receive probiotic prophylaxis. Three months after completed 7-day course of antimicrobial therapy, uropathogen bacteria were more often detected in vagina of women who did not receive probiotic prophylaxis (in 44 out of a total of 50 monitored patients - 88%) than in women who did not receive probiotic prophylaxis (in 18 out of a total of 53 patients - 34%). The results of this study have shown that prophylactic use of mixed probiotic culture present in medicinal product Acidosalus® in women with recurrent urinary tract infections is efficient and safe. We would like to stress the importance of simultaneous oral and vaginal application, by which we achieve regeneration and improved function of autochthonous intestinal and vaginal microflora, reduce a reservoir of uropathogen bacteria in intestines and boost immunity in general.

In the female body there are a vast number of skin changes that occur during pregnancy. Some of them are quite distressing to many women ; hereby is a discussion about performing treatment for physiological skin changes during pregnancy: antiinfective agents, glucocorticosteroids, topical immunomodulators, retinoids, minoxidil, etc. Drug administration during pregnancy must be reasonable.

The role of Ureaplasma urealyticum in bacterial prostatitis has not been completely clarified and there are no widely accepted criteria for defining prostatitis caused by these or other infrequently isolated pathogens. However, Ureaplasma urealyticum and other unusual pathogens found in expressed prostatic secretion (EPS)/urine voided immediately after prostatic massage (VB3) of some patients cannot be ignored. Also, Ureaplasma urealyticum found in EPS and VB3 has also been brought into connection to genesis of urolithiasis, infertility and Reiter's syndrome. The aim of this prospective study was to investigate the role of Ureaplasma urealyticum in chronic prostatitis.