Bacterial diversity across four drinking water distribution systems in Croatia: impacts of water management practices and disinfection by-products.

Several factors may impact bacterial diversity in drinking water distribution systems (DWDSs) including the origin of the raw water, the water treatment technologies, and the disinfection practices applied. 16S rRNA metabarcoding was used for the in-depth characterization of bacterial communities in the four studied Croatian DWDSs (A, B, C, D) two of which had residual disinfectant (A, B) and two were without (C, D), while only B utilized the conventional water treatment technology. Significantly higher diversity and species richness were evidenced in non-disinfected DWDSs (p<0.05) compared to disinfected DWDSs. The phylum Proteobacteria was the most abundant in all the DWDSs, being proportionately higher in non-disinfected systems (p<0.05). The most abundant genera in DWDS-A Mycobacterium and Sphingomonas both positively correlated, whereas Lactobacillus negatively correlated with the concentration of disinfection by-products (DBPs) as a sum of haloacetic acids (HAAs). Conversely, the genus Ralstonia positively correlated with the individual DBP dichloroacetic acid. These results indicate that genera Sphingomonas, Mycobacterium, Lactobacillus and Ralstonia could have an effect on promoting the formation of DBPs, in a similar manner to how negatively correlated taxa may influence their degradation.

In vitro and in vivo correlation of skin and cellular responses to nucleic acid delivery.

Skin, the largest organ in the body, provides a passive physical barrier against infection and contains elements of the innate and adaptive immune systems. Skin consists of various cells, including keratinocytes, fibroblasts, endothelial cells and immune cells. This diversity of cell types could be important to gene therapies because DNA transfection could elicit different responses in different cell types. Previously, we observed the upregulation and activation of cytosolic DNA sensing pathways in several non-tumor and tumor cell types as well in tumors after the electroporation (electrotransfer) of plasmid DNA (pDNA). Based on this research and the innate immunogenicity of skin, we correlated the effects of pDNA electrotransfer to fibroblasts and keratinocytes to mouse skin using reverse transcription real-time PCR (RT-qPCR) and several types of protein quantification. After pDNA electrotransfer, the mRNAs of the putative DNA sensors DEAD (AspGlu-Ala-Asp) box polypeptide 60 (Ddx60), absent in melanoma 2 (Aim2), Z-DNA binding protein 1 (Zbp1), interferon activated gene 202 (Ifi202), and interferon-inducible protein 204 (Ifi204) were upregulated in keratinocytes, while Ddx60, Zbp1 and Ifi204 were upregulated in fibroblasts. Increased levels of the mRNAs and proteins of several cytokines and chemokines were detected and varied based on cell type. Mouse skin experiments in vivo confirmed our in vitro results with increased expression of putative DNA sensor mRNAs and of the mRNAs and proteins of several cytokines and chemokines. Finally, with immunofluorescent staining, we demonstrated that skin keratinocytes, fibroblasts and macrophages contribute to the immune response observed after pDNA electrotransfer.

Mixing of arsenic-rich groundwater and surface water in drinking water distribution systems: Implications for contaminants, disinfection byproducts and organic components.

The utilization of groundwaters containing high levels of arsenic (As) for drinking water purposes presents major health and economic challenges for water utilities. One low-cost approach is to mix arsenic-rich groundwater (GW) with arsenic-free surface waters (SW) to achieve acceptable As levels. In this study we investigated the effect of different mixing ratios on water quality in an eastern Croatian water distribution system (WDS). To investigate the effects of mixing on drinking water quality, we measured the organic matter (OM) composition, disinfection byproduct (DBP) and metal concentrations in differently mixed ratios of GW and SW within the WDS. Fluorescence analysis revealed that the GW and SW had similar OM composition, with an almost equal ratio of humic- and protein-like OM throughout the WDS despite fluorescence indices revealing slightly different OM sources between the two water types. The tyrosine-like OM component was more variable, increasing during warmer months and towards the end of the WDS, most likely due to enhanced biofilm formation. Arsenic concentrations decreased to below 10 µg/L in the second half of the sampling campaign. Acceptable water quality was achieved after a period of destabilization and solubilization of loose deposits within the WDS resulting in their mobilization caused by water quality changes. Principal component and classification analysis, regression models and Spearman correlation coefficients revealed an association between As, OM and DBP concentrations with these correlations suggestive of their role in As mobilization in the WDS. Changing source waters, with different OM content and characteristics, corresponded to variable As release within the WDS.

Gene electrotransfer of IL-2 and IL-12 plasmids effectively eradicated murine B16.F10 melanoma.

Gene therapy has become an important approach for treating cancer, and electroporation represents a technology for introducing therapeutic genes into a cell. An example of cancer gene therapy relying on gene electrotransfer is the use of immunomodulatory cytokines, such as interleukin 2 (IL-2) and 12 (IL-12), which directly stimulate immune cells at the tumour site. The aim of our study was to determine the effects of gene electrotransfer with two plasmids encoding IL-2 and IL-12 in vitro and in vivo. Two different pulse protocols, known as EP1 (600 V/cm, 5 ms, 1 Hz, 8 pulses) and EP2 (1300 V/cm, 100 µs, 1 Hz, 8 pulses), were assessed in vitro for application in subsequent in vivo experiments. In the in vivo experiment, gene electrotransfer of pIL-2 and pIL-12 using the EP1 protocol was performed in B16.F10 murine melanoma. Combined treatment of tumours using pIL2 and pIL12 induced significant tumour growth delay and 71% complete tumour regression. Furthermore, in tumours coexpressing IL-2 and IL-12, increased accumulation of dendritic cells and M1 macrophages was obtained along with the activation of proinflammatory signals, resulting in CD4 + and CD8 + T-lymphocyte recruitment and immune memory development in the mice. In conclusion, we demonstrated high antitumour efficacy of combined IL-2 and IL-12 gene electrotransfer protocols in low-immunogenicity murine B16.F10 melanoma.

Effects of changing supply water quality on drinking water distribution networks: Changes in NOM optical properties, disinfection byproduct formation, and Mn deposition and release.

Field studies were conducted in a Croatian city supplied by two distinct groundwater sources (referred to as A and B) to investigate both the effects of changing water source on the water quality in the drinking water supply system, as well as to further understand discoloration events that occurred in city locations that switched water from source A to B. The water treatment processes at site A were found to alter organic matter (OM) characteristics, removing humic substances while enhancing protein-derived (tryptophan) content. Although the humic-like component predominated in raw waters, microbially/protein-derived components were found to increase throughout the distribution networks of both systems. Disinfection byproducts (DBPs) such as total trihalomethane (TTHM) and total haloacetic acid (THAA) were prevalent in water distribution system (WDS)-A, which correlated with elevated OM content as well as re-chlorination with hypochlorite (NaOCl). Our field study revealed that THMs were more readily formed than HAAs during ClO2 treatment. Unsurprisingly, chlorite concentrations were generally higher than chlorate concentrations during ClO2 treatment, whereas (secondary) NaOCl disinfection contributed to higher chlorate production. Principal component analysis indicated that variable pH values and humic-like OM could affect Mn, As and Al concentrations at the consumer's tap. Our results suggested that although Mn concentrations complied with regulations at WDS-B and were below 50 µg/L after disinfection, Mn was oxidized and formed particulate Mn oxides capable of causing discoloration events depending on prevailing network physico-chemical and hydraulic conditions. Aluminium also appears to be released during hydraulic disturbances from extensive deposits within the network. Thermodynamic calculations showed that Mn-oxidation was strongly dependent upon the ORP, and to lesser extent the pH value. Collectively, our results confirm that ensuring the provision of safe drinking waters to consumers requires an understanding of water quality across entire distribution networks in addition to any routine post-treatment monitoring.

Disinfection by-products in Croatian drinking water supplies with special emphasis on the water supply network in the city of Zagreb.

The occurrence of disinfection by-products (DBPs) was investigated in 48 drinking water systems across Croatia. Eleven DBPs were studied: chlorite (ClO2-), chlorate (ClO3-), four trihalomethanes (THMs), and five haloacetic acids (HAAs). Furthermore, an intensive sampling program was conducted in the distribution system in the city of Zagreb where, aside from DBP analyses, natural organic matter (NOM) was characterized using fluorescence spectroscopy. In the drinking waters examined across Croatia, DBP values were found in the range from 0.7 µg/L to 32.8 µg/L for THMs, below LOQ to 17.2 µg/L for HAAs (primarily di- and trichloroacetic acids), below LOQ to 720 µg/L for ClO2- and below LOQ to 431 µg/L for ClO3-. The results obtained showed higher chlorite concentrations in the systems treated with hypochlorite compared to systems treated with chlorine dioxide. DBPs in the Zagreb distribution network were generally low (the average values were below 6 µg/L and 2 µg/L for total THM and total HAA respectively). In contrast to our observations throughout Croatia, dibromoacetic acid (DBAA) was found to be the predominant HAA within Zagreb, most likely due to the degradation of chlorinated carboxylates (di-/tri-chloroacetic) in the network. Characterization of NOM by Parallel Factor Analysis (PARAFAC) fluorescence spectroscopy across the Zagreb network showed distinct temporal variations arising from groundwater inputs, as evident from variable humic-, tyrosine-, and tryptophan-like peaks. Statistical correlations between fluorescence data and DBPs highlight its potential for monitoring the presence of DBPs in distribution networks.

Pregabalin versus sertraline in generalized anxiety disorder. An open label study.

OBJECTIVE: Generalized Anxiety Disorder (GAD) is a chronic mental illness with a prevalence of 5-7% in the general population. t GAD is characterized by extreme persistent worry, mostly about minor problems, involving pathological fear with high occurrences of vegetative disturbance. GAD leads to functional impairment and a significantly reduced patient's quality of life. According to the guidelines of the World Federation of Societies of Biological Psychiatry (WFSBP), the first-line treatments for GAD are Serotonin selective reuptake inhibitors (SSRIs), Selective serotonin- and norepinephrine reuptake inhibitors (SNRIs) and pregabalin, an atypical anxiolytic. In this study, both efficacy and tolerability of pregabalin were evaluated and compared with efficacy and tolerability of sertraline, an SSRI antidepressant. PATIENTS AND METHODS: 107 patients both male and female, aged 20-60 were included in the study. All patients were hospitalized outward at the Psychiatric Clinic. Patients fulfilled criteria for GAD, according to ICD-X and DSM-IV. Each patient was randomly assigned to 4 weeks of treatment with pregabalin (n=47) or sertraline (n=60). Patients treated with sertraline were previously treated with SSRIs and SNRIs without remission, according to the latest National Clinical Guideline issued by the National Institute of Health and Clinical Excellence for treating GAD (NICE). The primary analysis was the change in the Hamilton Rating Scale for Anxiety (HAMA), a total score from baseline to endpoint. The second indicator of efficacy was the change in the HAMA psychic (emotional) and somatic (physical) scores, weekly, till endpoint. Global clinical assessment was conducted by using the Clinical Global Impression change rating (CGI). RESULTS: Both pregabalin and sertraline showed good results in treating symptoms of Generalized Anxious Disorder. The onset of action was shorter in treatment with pregabalin compared to the treatment with sertraline. In the patients treated with sertraline, the anxiolytic effect was detectable after at least 14 days while pregabalin showed initial good results during the first week of treatment. Adverse effects were reported in 28% patients treated with pregabalin and 27% of patients treated with sertraline, without significant differences. There were no drop-out patients in neither group. Beside pharmacotherapy, each patient received 8 weeks of cognitive/behavior therapy. In concomitant therapy benzodiazepine was used (klonazepam, in low doses). In all patients adverse events were short-lasting withmild intensity and there were no withdrawal events during this study. CONCLUSIONS: Efficacy and tolerability of pregabalin were high. Compared to sertraline, pregabalin showed more rapid onset of action and equal efficacy. Adverse reactions are short-lasting and the dose depends. Our investigation showed that pregabalin, an atypic anxiolytic is efficient and well tolerable in treatment of GAD.

Gene electrotransfer of plasmid AMEP, an integrin-targeted therapy, has antitumor and antiangiogenic action in murine B16 melanoma.

Gene therapy with Plasmid AMEP (antiangiogenic metargidin peptide) has recently been studied as a potential targeted therapy for melanoma. This plasmid is designed to downregulate α5ß1 and αvß3 integrins. In our study, electroporation was used as a nonviral delivery system. We investigated the antiangiogenic and direct antitumor effectiveness of this gene therapy on low and highly metastatic B16 melanoma variants. In vitro, the antiangiogenic effectiveness as determined by tube formation assay on endothelial cells was predominantly dependent on AMEP expression levels. In vivo, antitumor effectiveness was mediated by the inhibition of proliferation, migration and invasion of melanoma cells and correlated with the expression of integrins on tumor cells after intratumor delivery. In addition, reduced metastatic potential was shown. Intramuscular gene electrotransfer of Plasmid AMEP, for AMEP systemic distribution, had no antitumor effect with this specific preventive treatment protocol, confirming that direct tumor delivery was more effective. This study confirms our previous in vitro data that the expression levels of integrins on melanoma cells could be used as a biomarker for antitumor effectiveness in integrin-targeted therapies, whereas the expression levels of AMEP peptide could be a predictive factor for antiangiogenic effectiveness of Plasmid AMEP in the treatment of melanoma.

Percutaneous closure of large femoral artery access with Prostar XL in thoracic endovascular aortic repair.

OBJECTIVES: To investigate the technical success rate of Prostar XL for closure of large (≥20F) femoral vascular access sites in thoracic endovascular aortic repair (TEVAR) procedures. METHODS: This was a single-center consecutive case series. All TEVAR procedures at Uppsala University Hospital 2006-2010 were registered prospectively. Reoperations and cases with open closure technique were excluded. Primary (early) technical failure was defined as closure failure requiring immediate (on-table) open surgical repair; late access-related complication occurred thereafter. The medical records, pre- and postoperative computed tomography images were reviewed retrospectively. RESULTS: A total of 164 TEVAR procedures were identified, of which 118 (71%) had a median 22F (range 20-26F) access site sealed with tandem Prostar XL. The indications for TEVAR were dissection (47%), aneurysm (42%), trauma (8%), and miscellaneous (3%). Median follow-up time was 10 months (range 1-62). Primary technical failure occurred in 10 of 118 (8%). These cases were converted to cut-downs and surgical repair (n = 7), femoral fascia suturing (n = 2), and external compression with the Femo-Stop device (n = 1). Hypertension was associated with primary failure (p = .005), and a trend was observed for high age (p = .078) and increased groin subcutaneous fat layer (p = .077). Late access-related complications included pseudo-aneurysms (n = 12), small hematomas (n = 7), superficial groin infections (n = 2), and deep venous thrombosis (n = 1). None of the late complications required surgical treatment. CONCLUSIONS: The access closure technique with tandem Prostar XL for large access sites during TEVAR is safe, in experienced hands. Few technical failures and few late complications occur, and they are usually benign.

Hydrochemical characterization of arsenic contaminated alluvial aquifers in Eastern Croatia using multivariate statistical techniques and arsenic risk assessment.

Multivariate statistical analyses, cluster, factor and discriminant analyses, were used to estimate spatial variations in groundwater chemistry in Eastern Croatia and to identify the main geochemical processes responsible for high arsenic (As) concentrations in the groundwater analyzed. Cluster analysis revealed five different groups of sampling sites linked with groundwater evolution, hydrochemical characteristics and different As content in the groundwater. Two-model factor analysis explained around 50% of total variance of the data sets and enabled identification of the different geochemical processes responsible for higher As concentrations, i.e. decoupled Fe and As reduction and desorption. Using discriminant analysis, a three-parameter discriminant function was derived: electrical conductivity, nitrate and bromide, which yielded highly accurate classification of the samples according to the concentration of As as As-safe (<10µg/L) and As contaminated (>10µg/L). A health risk assessment model was applied to calculate cumulative exposure to As as well as toxic and carcinogenic risks resulting from drinking raw groundwater contaminated by As in Eastern Croatia. Although the results obtained indicate that adverse health effects could be observed among the residents of the villages in which raw groundwater with higher As concentrations has been used, there are no reported cases of arsenicosis.

[Depression in older persons]. / Depresije u osoba starije zivotne dobi.

INTRODUCTION: Depression is a common problem in old age, especially among those with various medical illnesses. Aging brings about numerous losses that might lead to depression: loss of health, loved ones and social roles. Some of older persons develop depression. Depression in old age is a serious illness. The suicide rate among older persons is very high. Approximately 90% of elderly people who commit suicide suffer from depression. Adequate treatment of depression in late life includes reducing and resolving depressive symptoms, preventing its relapse and reoccurrence, improving quality of life, and reducing mortality and healthcare costs. MATERIAL AND METHODS: 32 female inpatients were examined. All patients suffered from depressive disorder (middle, according to International Classification of Diseases (ICD-X). Only patients without cognitive impairment were included in the study, according to Mini-Mental State Examination. Hamilton Rating Scale for Depression was used as primary measure of efficacy. Patients were evaluated for efficacy weekly. For statistical analysis t test was used. RESULTS: Depressive disorders are more severe among older patients, with greater suicidal risk, anxiety and psychomotor agitation. Patients treated with moclobemide show quicker remission of depressive symptoms compared to patients who were treated with mianserin. CONCLUSION: Depressive disorders among older patient are very serious illnesses, but with adequate therapy, symptoms can disappear, and there is no difference between remission in depression among older and younger patients.

[Comparison of the efficacy of traditional antidepressive agents and the new generation of antidepressives in the treatment of depressive disorders]. / Komparacija efikasnosti tradicionalnih antidepresiva i antidepresiva nove generacije u lecenju depresivnih poremecaja.

INTRODUCTION: Considerable efforts have been made throughout the last 20 years to develop drugs that can replace tricyclic antidepressants as the primary treatment for depression. These efforts are well justified since tricyclic antidepressants, although therapeutically quite efficient, cause several problems, such as side effects and toxicity at therapeutic doses, causing particular problems in the elderly and in patients with congestive heart disease, and giving severe toxicity when taken in overdose. A number of compounds that are pharmacodynamically different from the tricyclics have been developed, and Selective Serotonin Reuptake Inhibitors are such. This group of antidepressives has been developed and widely marked as antidepressant therapy which offers safety and tolerability advantages over tricyclics. Tricyclic antidepressants have been the standard drug treatment for depressive illness, against which the efficacy of new compounds is compared in this investigation. MATERIAL AND METHODS: 30 female inpatients were examined. All patients had to fulfill the inclusion criteria for moderate depressive disorders, according to International Classification of Diseases. The primary efficacy parameters in the study were the changes from baseline Hamilton Rating Scale for Depression with 19 items, total score at endpoint. Patients were evaluated weekly for efficacy and adverse events. Clinicians also rated patients on the Hamilton Rating Scale for Anxiety. In statistical analysis X and T test were used. RESULTS: There was no significant difference between two groups of patients in response to tricyclic antidepressants and selective serotonin reuptake inhibitors. Selective Serotonin Reuptake Inhibitors show less adverse effects compared to tricyclics, so they appeared to be better tolerated. CONCLUSION: The traditional and new antidepressive drugs appeared to be equally effective in the treatment of moderate depressive disorders, but new drugs: Selective Serotonin Reuptake Inhibitors show less adverse effects, compared to the traditional, tricyclic antidepressants.

[Clinical characteristics of unipolar and bipolar depression]. / Klinicke karakteristike unipolarnih i bipolarnih depresija.

INTRODUCTION: In the last decades affective disorders were divided into unipolar and bipolar and this division has been generally accepted. The bipolar type is manifested by mania or by both mania and depression. On the other hand, unipolar affective disorders are manifested only by depression. In numerous investigations authors have noticed that there are very distinctive differences between these two types of depressive disorders such as: course of illness, personality disorders, sex, family history etc. Nevertheless, in practice it is often very difficult to make the right diagnosis. The bipolar type often starts with a few pure depressive episodes and sometimes mania occurs a few years later so only at that point the psychiatrist can make the right diagnosis and treat the patient correctly. MATERIAL AND METHODS: This investigation comprised 50 patients hospitalized at the Psychiatric Clinic in Novi Sad during 1992-1995. The experimental group consisted of 20 patients with a bipolar affective disorder (according to ICD-X), while the control group consisted of 30 patients with clinical diagnosis of unipolar depression (intensive, without psychiatric features). Both groups of patients were weekly evaluated by Hamilton Depression Rating Scale (HDRS), whereas the initial score for all patients had to be higher than 16. RESULTS: Patients suffering from unipolar depression were older than patients with bipolar depression and there were more females in this group. There were no differences in demographic characteristics (level of education, migration, etc.), but the experimental group had a greater genetic loading for affective disorders. Unipolar depressive patients had more agitation and they were more anxious than patients with bipolar depression. DISCUSSION AND CONCLUSION: The fact that unipolar depressive patients were older than bipolar is similar to most of the results gained in this kind of investigation. On the other hand, we did not find statistical differences in the intensity of disorders, and in the literature these results are contraindicating. Numerous investigators report that bipolar depressives had a stronger genetic loading for affective disorders and our study confirms the same. All these results can help us to make the right diagnosis of unipolar and bipolar affective disorders.

[Clinical characteristics of depressive disorders]. / Klinicke karakteristike depresivnih poremecaja.

In this article authors investigated the clinical distinction between endogenous and neurotic depression. The endogenous depression was equivalent to severe depressive episode without psychotic symptoms, according to ICD-X criteria, and neurotic depression was equivalent to dysthymia. The results showed that endogenous depressive patients were much older than neurotic depressives, and that they had more severe symptoms of depression. On the other hand endogenous depressive patients responded better to antidepressive therapy, and consequently they had better prognosis than neurotic depressive patients.

Duplex scanner versus conventional arteriography for carotid artery disease evaluation.

In order to evaluate extracranial circulation, Duplex scanning and conventional angiography were performed. The group of 41 patients (mean age 53.8 +/- 11.8, 71% male) were examined. Results were analyzed for extracranial disease presence such as presence of vessel stenosis (mild, moderate, severe), occlusion and presence of plaques. Data from Duplex scanning and carotid arteriography were correlated within the patients in the term of sensitivity and specificity of the methods used. The sensitivity of Duplex scanning for disease presence was 93% and specificity was 83%. For the presence of stenosis, sensitivity was 79% and specificity was 62%. For mild and moderate stenosis (less than 50% of lumen narrowing) sensitivity was 63%, whereas for severe, high-grade stenosis (greater than 50%) and occlusion the sensitivity was 100%. For the plaque presence the sensitivity was 52% and specificity was 73%. A good accordance of plaque findings with both methods was found only in 24% of patients. By Duplex scanner the plaques were discovered in 92% of patients while by arteriography they were found only in 32%. Duplex scanning and conventional arteriography are both accurate methods for determination of the reduction in diameter of carotid arteries (especially in stenosis greater than 50%), whereas Duplex scanning is more accurate for plaque detection and plaque characterisation.