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Background: Optimized and efficient library preparation workflow is one of the most important prerequisites for obtaining high quality and quantity of results in massively parallel sequencing (MPS). Our aim was to assess and optimize different steps of Illumina® Nextera® XT assay for analysis of whole mitochondrial genomes.Methods and Results: Among the three long-range high-fidelity DNA polymerases tested here, PrimeSTAR® GXL performed best in aspects of specificity and yield for mitochondrial DNA (mtDNA) enrichment. Furthermore, library quantification combined with individual library-by-library dilution outperformed bead-based normalization in terms of more equal distribution of reads per library, reduced hands-on time and simplified workflow. Increasing the number of amplification cycles in the index-adapters-adding PCR step had no adverse effect on the level of sequencing noise, which remained low both in negative controls and in samples.Conclusions: Optimizations described herein provide beneficial insights for laboratories aiming at implementation and/or advancement of similar MPS workflows (e.g. small genomes, PCR amplicons and plasmids).

The aim of the study was to assess forensic pertinence of 12 short tandem repeats (STRs) on X-chromosome in south Croatia population. Investigator® Argus X-12 kit was used to co-amplify 12 STR loci belonging to four linkage groups (LGs) on X-chromosome in 99 male and 98 female DNA samples of unrelated donors. PCR products were analyzed by capillary electrophoresis. Population genetic and forensic parameters were calculated by the Arlequin and POPTREE2 software, and an on-line tool available at ChrX-STR.org. Hardy–Weinberg equilibrium was confirmed for all X-STR markers in female samples. Biallelic patterns at DXS10079 locus were detected in four male samples. Polymorphism information content for the most (DXS10135) and the least (DXS8378) informative markers was 0.9212 and 0.6347, respectively. In both male and female samples, combined power of discrimination exceeded 0.999999999. As confirmed by linkage disequilibrium test, significant association of marker pair DXS10074-DXS10079 (P = 0.0004) within LG2 and marker pair DXS10101-DXS10103 (P = 0.0003) within LG3 was found only in male samples. Number of observed haplotypes in our sample pool amounted 3.01, 7.53, 5 and 3.25% of the number of possible haplotypes for LG1, LG2, LG3 and LG4, respectively. According to haplotype diversity value of 0.9981, LG1 was the most informative. In comparison of south Croatia with 26 world populations, pair-wise $${{\text{F}}_{\text{ST}}}^{\text{*}}$$ values increase in parallel with geographical distance. Overall statistical assessment confirmed suitability of Investigator® Argus X-12 kit for forensic casework in both identification and familial testing in the population of south Croatia.

The most recent data on the incidence of brucellosis in Southeast Europe prove the persistence of this zoonosis in the area, regardless of constant efforts at controlling it as one of the most dangerous zoonoses. Forty-three Brucella melitensis strains were collected from cattle, sheep, goats and humans from Croatia as well as Bosnia and Herzegovina between 2009 and 2015. The strains were identified and genotyped in order to determine their epidemiological background. Standard biotyping methods and Bruce-ladder were used to identify the strains. Genotyping was done using multilocus variable number tandem repeats analysis (MLVA) on 16 and multilocus sequence typing analysis (MLST) on nine loci. Results were compared to each other and to internationally available data. Twenty- five novel genotypes and two sequence types were identified. All tested strains, apart from vaccine and reference strains, showed very close phylogenetic and geographic relationships. The genotyping results indicate the endemicity of brucellosis in this region. MLST showed no variation, confirming the stability of housekeeping genes. The results confirm already established routes of disease spread in this area, showing that a more detailed and vigorous control of this zoonosis is necessary.

In order to implement X-chromosome short tandem repeat (X-STR) typing into routine forensic practice, reference database of a given population should be established. Therefore we extended already published data with additional 397 blood samples from unrelated Croatian citizens, and analyzed the total of 995 samples (549 male and 446 female) typed by Investigator® Argus X-12 Kit. To test genetic homogeneity of consecutively processed five historic-cultural regions covering the entire national territory, we calculated pairwise Fst genetic distances between regions based on allele and full haplotype frequencies. Since the comparison did not yield any statistically significant difference, we integrated STR profile information from all regions and used the whole data set to calculate forensic parameters. The most informative marker is DXS10135 (polymorphism information content (PIC = 0.929) and the most informative linkage group (LG) is LG1 (PIC = 0.996). We confirmed linkage disequilibrium (LD) for seven marker pairs belonging to LG2, LG3 and LG4. By including LD information, we calculated cumulative power of discrimination that amounted to 0.999999999997 in females and 0.999999005 in males. We also compared Croatia with 13 European populations based on haplotype frequencies and detected no statistically significant Fst values after Bonferroni correction in any LG. Multi- dimensional scaling plot revealed tight grouping of four Croatian regions amongst populations of southern, central and northern Europe, with the exception of northern Croatia. In this study we gave the first extensive overview of aberrant profiles encountered during Investigator® Argus X-12 typing. We found ten profiles consistent with single locus duplication followed by tetranucleotide tract length polymorphism. Locus DXS10079 is by far the most frequently affected one, presumably mutated in eight samples. We also found four profiles consistent with X-chromosome aneuploidy (three profiles with XXX pattern and one profile with XXY pattern). In conclusion, we established integral forensic Croatian X- chromosome database, proved forensic pertinence of Investigator® Argus X-12 Kit for the entire Croatian population and identified locus DXS10079 as a potential duplication hotspot.

Although Q fever affects humans and animals in Croatia, we are unaware of genotyping studies of Croatian strains of the causative pathogen Coxiella burnetii, which would greatly assist monitoring and control efforts. Here 3261 human and animal samples were screened for C. burnetii DNA by conventional PCR, and 335 (10.3%) were positive. Of these positive samples, 82 were genotyped at 17 loci using the relatively new method of multi-locus variable number tandem repeat analysis (MLVA). We identified 13 C. burnetii genotypes not previously reported anywhere in the world. Two of these 13 genotypes are typical of the continental part of Croatia and share more similarity with genotypes outside Croatia than with genotypes within the country. The remaining 11 novel genotypes are typical of the coastal part of Croatia and show more similarity to one another than to genotypes outside the country. Our findings shed new light on the phylogeny of C. burnetii strains and may help establish MLVA as a standard technique for Coxiella genotyping.

Summary Anthrax is a serious disease caused by Bacillus anthracis. Humans can become infected by handling products from infected animals, by breathing spores and rarely by eating undercooked meat from infected animals. The genome of B. anthracis is highly monomorphic and thus shows very low DNA sequence variation. We analysed the molecular characteristics of 12 B. anthracis isolates from outbreaks in Croatia and Bosnia and Herzegovina, which have occurred during the past 10 years along with two vaccine strains. Genotyping system based on variable-number tandem repeat analysis at six loci revealed that six isolates belong to genotype from the A1.a cluster whilst six isolates relate to the B2 cluster, compared to 89 previously described genotypes. The distribution of two evolutionarily distant clusters suggests an introduction of B. anthracis to this area in at least two separate events.

Bacillus anthracis can infect both livestock and humans. The importance of the treatment of the disease in humans has been underscored by the bioterrorism events of 2001 in the United States. The presence of PA and B/C genes (pX01 and pX02 plasmids) as well as susceptibility to several antimicrobial substances was determined in 11 strains of Bacillus anthracis isolated during two recent epizooties of anthrax which occurred in Croatia in 2002 among sheep and in 2006/2007 in cattle. The pX01 plasmid was observed in all of the examined strains, including vaccinal Sterne strains. However, the pX02 plasmid was detected in only eight out of eleven examined field strains of anthracis while in vaccinal strains it was not detected at. Determination of MIC's revealed susceptibility to amoxicillin, amoxicillin with clavulanic acid, ciprofloxacin, gentamicin and tetracycline. All strains were resistant to sulfamethoxazole with trimethoprim and cefotaxime.

Aim To present the surveillance data on Brucella melitensis, B. suis, and B. ovis infection in cattle, sheep, goats, and swine in Croatia obtained in 2008 by serological, bacteriological, and molecular methods for diagnostics of brucellosis in domestic animals. Methods We serologically tested 42 785 cattle serums, 22 686 sheep and goat serums, and 28 520 swine serums using the Rose Bengal test, complement fixation test, and various immunosorbent assays. We also tested 10 173 ram blood samples for B. ovis infection using the complement fixation test. Bacteriological examination was conducted on 214 samples collected from 34 serologically positive animals. Different molecular methods were employed in the identification and typing of 20 isolates from the samples. Results B. melitensis biovar (bv.) 3 was confirmed with different identification methods in 2 flocks in 2 Croatian counties and B. suis bv. 2 in 3 flocks in 3 counties. B. melitensis in cows was confirmed for the first time in Croatia. Infection with B. ovis was serologically confirmed in 202 rams in 12 counties. Conclusions In 2008, the size of the brucellosis-affected area in Croatia and the efficiency of detection and prevention of brucellosis in sheep, goats, and swine were satisfactory. Infection with B. melitensis in cattle was confirmed for the first time and possible links for infection in humans were detected. More efficient measures for suppression and control of ovine epididymitis are required and a new strategy may be necessary for complete eradication of this disease.

Mycobacterial spindle cell ‘pseudotumour’ has been described only once in cats. This unique proliferation of spindle-shaped histiocytes containing Mycobacterium avium is associated with extensive subcutaneous lesions. We report mycobacterial pseudotumour with invasion of muscular and subcutaneous tissues in a 1-year-old female domestic longhair cat. Lesions involved the facial muscles and nasal cavity, making surgical excision impossible. Necropsy examination revealed additional nodules in the subcutis and muscles of the trunk and submandibular lymph nodes. Genotyping of organisms within these lesions revealed simultaneous infection with Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis. The microscopical appearance of the granulomas was identical, regardless of the strain of bacterium or anatomical location.

The association of interferon (IFN) therapy with haemolytic uraemic syndrome in patients with chronic myeloid leukaemia (CML) has been reported infrequently. The pathogenesis of the renal lesion in such cases remains unclear. We report the case of a patient with chronic myeloid leukaemia who developed nephrotic syndrome and renal failure while being treated with hydroxyurea and IFN-alpha. Renal biopsy showed features of chronic thrombotic microangiopathy. The discontinuation of IFN-alpha, and a prompt institution of plasmapheresis and steroids resulted in improvement of the nephrotic syndrome and renal function. These findings suggest that long-term IFN-alpha therapy can induce thrombotic microangiopathy and haemolytic uraemic syndrome in patients with chronic myeloid leukaemia.

In the period from 2009 to 2013, bacteriological testing was conducted on 336 sheep, goat and cattle samples. Using classical bacteriological and molecular procedures, B. melitensis was confirmed in 14 (4.2 %) samples. Brucella was isolated in the Karlovac, Lika-Senj and Split-Dalmatia Counties. Brucella isolates were genotyped using the MLVA method and compared with isolates from neighbouring Bosnia and Herzegovina (BH). A total of 14 isolates (strains) originating from Croatia and 25 from BH were analysed. Complete matches between Croatian and BH isolates were found in two genotype groups. Overall, the MLVA analysis indicated that the Croatian and BH genotypes of B. melitensis from animals were highly homogenous. The Hunter Gaston diversity index (HGDI) showed that diversity was found among the 16 tested loci for 5 loci of panel 2B (Bruce 04, 07, 09, 16, 30). The discriminating loci are optimal for the use in epidemiological investigation of B. melitensis infections in these two countries., U razdoblju od 2009. do 2013. u Hrvatskoj je izvršena bakteriološka pretraga uzoraka ukupno 336 ovaca, koza i goveda. Klasičnim bakteriološkim i molekularnim postupcima B. melitensis je potvrđena u 14 (4,2%) izolata. Brucele su izdvojene u Karlovačkoj, Ličko-senjskoj i Splitsko-dalmatinskoj županiji. Analiza broja uzastopnih ponavljanja na više lokusa (engl. multiple locus variable number of tandem repeat analysis, MLVA) primijenjena je radi genotipizacije i usporedbe s izolatima iz susjedne Bosne i Hercegovine. Ukupno je analizirano 14 izolata podrijetlom iz Hrvatske i 25 iz susjedne Bosne i Hercegovine. U dva slučaja genotipovi izolata iz Hrvatske u potpunosti su se poklapali s genotipovima u Bosni i Hercegovini. Korištenjem metode MLVA utvrđen je visok stupanj homogenosti izolata u ove dvije zemlje. Od korištenih 16 MLVA lokusa raznolikost je utvrđena samo na 5 lokusa panela 2B (Bruce 04, 07, 09, 16, 30) što je potvrđeno računanjem HGDI indeksa. Korištenje ovih 5 razlikovnih lokusa optimalno je za provođenje epidemioloških istraživanja u Hrvatskoj i Bosni i Hercegovini.

Porcine brucellosis is a common bacterial zoonosis which can cause significant financial losses. Its diverse and often complicated factors have hampered efforts to control disease spread. The aim of the study was to assess the epidemiological situation of porcine brucellosis primarily in Croatia and its relationship to genotypes present in other, mostly European countries. One hundred and seven Brucella suis strains isolated from swine, hares, cattle, humans, wild hares, a wild boar and a mare originating mainly from Croatia (112), but also a few from Slovenia, Bosnia and Herzegovina, Serbia and Macedonia (15) were tested using classical microbiological testing, Bruce-ladder, RFLP, Multiplex-suis and genotyped using multi-locus variable-number tandem repeat analysis (MLVA). We determined 43 Brucella suis genotypes. Strains were grouped according to phylogenetic and geographic relationships, revealing both regional specificity and uniqueness and suggesting possible sources and modes of spread among animals. Our study also confirmed problems with Bruce19 locus that may hinder comparisons of new types with those in the international database. Forty-one novel genotypes were identified and deposited into the international database. Our study supports the idea of wild animals as a source of disease in domestic animals and also gives evidence to hypothesis of cross-border animal trafficking between former Yugoslavian countries. It also highlights the need to expand such research across more of southeast Europe, especially to countries with poorer social and economical situation in order to prevent a realistic outbreak and for better understanding of the biology of this pathogen.

The aim of this study was to determine the presence of enterotoxin genes in coagulase - negative staphylococci from traditional Croatian fermented sausages. A total of 39 isolates were collected from the dry sausage “kulen” and Slavonian homemade sausages as presumptive CNS, and were subjected to PCR identification to the genus level. All isolates belonged to the Staphylococcus genus. Strains were tested for the presence of 13 enterotoxin genes; seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN and seO by the PCR method. All strains were negative to all enterotoxin genes. The observed results indicate the absence of risk related to enterotoxogenic potential of coagulase - negative staphylococci as indigenous microbiota from autochthonous Croatian traditional sausages., Cilj ovog rada bio je utvrditi prisutnost gena za enterotoksine u koagulaza - negativnih stafilokoka iz tradicionalnih hrvatskih fermentiranih kobasica. Ukupno je prikupljeno 39 izolata iz kulena i domaće slavonske kobasice te identificirano PCR - om do razine roda. Svi izolati pripadali su rodu Staphylococcus. PCR - om je određivana prisutnost 13 gena za enterotoksine: seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN i seO. Ni u jednom izolatu nije utvrđena prisutnost gena za enterotoksine. Dobiveni rezultati upućuju na odsutnost rizika povezanog s toksogenim potencijalom koagulaza-negativnih stafilokoka kao dijela autohtone mikroflore hrvatskih fermentiranih kobasica.

Introduction: Skin granulomas of cats are rarely caused by saprophytic mycobacteria from Mycobacterium avium complex (MAC) which are cultivable. MAC skin infection has unspecific cutaneous nodular presentation and three histomorphologic forms: granulomatous, pyogranulomatous and spindle cell. We represent four cases of MAC induced skin granulomas in cats originating from seaport of island Hvar. Materials and Methods: In three cases submitted as skin neoplasia in cats we diagnosed granulomatous inflammation and presumed feline lepromatous leprosy based on granuloma architecture. In fourth case feline leprosy was suspected based on impression smear finding of negatively stained intracellular bacilli taken from ulcerated nodule. Mycobacterial culture was performed from granuloma tissue of two cats. DNA was extracted from formalin fixed paraffin embedded tissue and from fresh tissue. Partial fragment of 16S rRNA was amplified and subsequently sequenced and compared with sequences in GenBank. Routine histology and Ziehl-Neelsen staining were performed in fourth case. Results: Mycobacterium organisms were not detected by culture after 12 weeks of cultivation. In first three cases granuloma architecture was rich in foamy macrophages and multinuclear giant cells harboring high amount of slender acid-fast bacilli. Organisms were numerous within spindle cells in fourth case of typical MAC spindle cell proliferation. All sequences were identical. They revealed 100% identity with sequences of MAC members. Conclusions: Mycobacteria growth performances on synthetic media, as well as histological pattern of skin granulomas are not sufficient to distinguish etiology highlighting molecular identification as golden standard.

Methicillin resistant Staphylococcus aureus (MRSA) have emerged worldwide and have became resistant to a variety of antibiotics. MRSA colonisation in pigs was first reported in the Netherlands at 2005, where pigs were implicated as a source of human MRSA infections. Dust samples were collected from six large pig farms in Croatia from breeding pig facilities. On each farm, between 5 and 6 samples were taken by swabs. Of the total 32 swabs, isolates from 8 swabs from four of the six examined farms grew pink colonies on the MRSA select agar. The mecA gene was detected in all 8 isolates growing pink colonies on the MRSA agar. All isolates are resistant to penicillin, ampicilin, oxacillin, oxitetracycline and streptomycin. All isolates were susceptible to vancomycin, ciprofloxacin, florfenicol and sulfametoxazole/trimethoprim., Meticilin-rezistentni sojevi bakterije Staphylococcus aureus (MRSA) izdvajaju se u cijelom svijetu i otporni su na različite antimikrobne lijekove. Infekcije svinja sojevima MRSA-e prvi su put opisane u Nizozemskoj 2005. s pretpostavkom da su svinje izvor infekcije MRSA-om za ljude. Uzorke prašine prikupljali smo sa šest velikih svinjogojskih farmi u Hrvatskoj iz objekata u kojima su smještene rasplodne životinje. Na svakoj farmi uzeli smo pet ili šest brisova. Od ukupno uzorkovana 32 brisa, iz osam brisova s četiri farme izdvojili smo ružičaste kolonije na MRSA selektivnom agaru. Kod svih izolata koji su rasli na MRSA selektivnom agaru kao ružičaste kolonije izdvojen je gen mecA. Svi izolati bili su otporni na penicilin, ampicilin, oksacilin, oksitetraciklin i streptomicin, a osjetljivi na vankomicin, ciprofloksacin, florfenikol i sulfametoksazol s trimetoprimom.

During the research period (2001-2005), the lymph nodes of 123 wild boars from seven locations in the Republic of Croatia were tested for the presence of mycobacteria. Mycobacteria were isolated from the lymph nodes of 15 (12.2%) wild boars from fifi ve locations. Specific hybridization showed that 11 (8.9%) isolates belonged to M. avium, 2 (1.6%) isolates were typed as M. fortuitum, and 2 (1.6%) isolates belonged to M. gordonae. All 11 isolates belonging to M. avium sp. were tested by means of the specififi c primers P1 FR300 and P2 FR300. Product size in all isolates was 300 bp, which confirms that M. avium subsp. hominissuis was present in all of them. As in swine from intensive farming, M. avium subsp. hominissuis is a dominant mycrobacteria type in wild boar in Croatia. The results lead to the conclusion that the most important source for wild boar is an environment suitable for the development and survival of M. avium subsp. hominissuis, as confirmed in earlier research studies by other authors., Tijekom istraživanog razdoblja (2001. do 2005.) radi pretrage na prisutnost mikobakterija obrađeni su limfni čvorovi 123 divlje svinje iz sedam lokaliteta u Republici Hrvatskoj. Mikobakterije su izdvojene iz limfnih čvorova 15 (12,2%) divljih svinja iz pet lokaliteta. Specifičnom hibridizacijom dokazano je da 11 (8,9%) izolata pripada vrsti M. avium sp., 2 (1,6%) izolata tipizirana su kao M. fortuitum, a 2 (1,6%) izolata pripadali su vrsti M. gordonae. Svih 11 izolata koji su pripadali M. avium sp. testirani su pomoću specifičnih početnica P1 FR300 i P2 FR300. U svih izolata veličina proizvoda iznosila je 300 parova baza što potvrđuje da se u svih radi o vrsti M. avium subsp. hominissuis. Kao i u svinja iz intenzivnog uzgoja, M. avium subsp. hominissuis predstavlja dominantnu vrstu mikobakterija u divljih svinja u Hrvatskoj. Na temelju dobivenih rezultata može se zaključiti da je po svemu sudeći najvažniji izvor za divlje svinje okoliš koji pogoduje razvoju i opstanku M. Avium subsp. hominissuis, što potvrđuju i ranija istraživanja drugih autora.

Methicillin-resistantStaphylococcus aureus(MRSA) have emerged worldwide and have become resistant to a variety of antibiotics. MRSA colonisation in pigs was first reported from the Netherlands in 2005, where pigs were implicated as a source of human MRSA infections (Voss et al., 2005). This paper presents the first report on the presence of MRSA on large pig breeding farms in Croatia, together with the determination of themecAgene, the results ofspatyping and susceptibility to commonly used antimicrobials. Dust samples (7–11 per farm) were collected from eight large pig farms in Croatia. Of the total 68 swabs, themecA gene was detected in 24 isolates growing on the MRSA agar. All isolates were resistant to oxacillin, tetracycline and streptomycin, and susceptible only to vancomycin, while 92% of the strains were susceptible to ciprofloxacin. Genotyping of the MRSA strains was performed byspatyping, and revealed t011 (n = 17), t034 (n = 5) and t1451 (n = 2). The results presented here predict that MRSA is present on a large number of pig farms in Croatia.

Udruženost dijabetične nefropatije s primarnim glomerulonefritisom rijetka je kod šećerne bolesti tip 1. Nekoliko izvješća pokazuje da se primarni glomerulonefritis može razviti na osnovi dijabetične nefropatije. Ovi glomerulonefritisi obuhvaćaju idiopatski membranski glomerulonefritis, IgA glomerulonefritis, Henoch-Schönleinov nefritis, membranoproliferacijski glomerulonefritis, lupus nefritis, bolest minimalne promjene, postinfekcijski glomerulonefritis i brzo progredirajući glomerulonefritis. Kako neke od ovih bolesti mogu promijeniti liječenje i prognozu bubrežne bolesti kod dijabetičnih bolesnika, patološki nalaz u mokraći ili pogoršanje bubrežne funkcije koji nisu u skladu s naravom dijabetične nefropatije ukazuju na mogućnost nedijabetične bubrežne bolesti i zahtijevaju podrobniju procjenu. Opisuje se bolesnik sa šećernom bolešću tip 1 kod kojega je napravljena biopsija bubrega zbog teške proteinurije i nefrotskog sindroma. Biopsija bubrega je pokazala dijabetičnu nefropatiju, ali i fokalnu segmentnu glomerulosklerozu. Prema našim saznanjima, ovo je prvi objavljeni slučaj primarne žarišne segmentne glomeruloskleroze udružene s dijabetičnom nefropatijom i dijabetičnom retinopatijom, s izvrsnim odgovorom na terapiju., The association of diabetic nephropathy with primary glomerulonephritis in type 1 diabetes is rare. Several reports have shown that primary glomerulonephritis can be superimposed on diabetic nephropathy. These glomerulonephritides include idiopathic membranous glomerulonephritis, IgA glomerulonephritis, Henoch-Schönlein nephritis, membranoproliferative glomerulonephritis, lupus nephritis, minimal change disease, postinfectious glomerulonephritis and rapidly progressive glomerulonephritis. Because some of these disorders can alter the management and prognosis of renal disease in diabetic patients, the appearance of urinary abnormalities or deterioration in renal function inconsistent with the natural history of diabetic nephropathy raises the possibility of a nondiabetic renal disease and should lead to a more detailed evaluation. We report on a patient with type 1 diabetes that underwent renal biopsy because of heavy proteinuria and nephrotic syndrome. Renal biopsy showed diabetic nephropathy coexistent with focal segmental glomerulosclerosis. To the best of our knowledge, this is the first reported case of primary focal segmental glomerulosclerosis associated with diabetic nephropathy and diabetic retinopathy, with excellent therapeutic response.

U radu je prikazana 31-godišnja bolesnica s hipokalijemijskom kvadriparezom. Dijagnosticirana je distalna renalna tubularna acidoza u sklopu primarnog Sjögrenovog sindroma. Rijetko je hipokalijemijska paraliza početna manifestacija primarnog Sjögrenovog sindroma. U naše bolesnice prethodila je simptomima kserostomije i kseroftalmije. Na dijagnozu primarnog Sjögrenovog sindroma treba pomišljati u mlađih žena sa simptomima progresivne slabosti, hipokalijemije i metaboličke acidoze., We report a case of hypokalaemic quadriparesis in 31-year old woman in whom the discovery of distal renal tubular acidosis led to the diagnosis of primary Sjögren’s syndrome. Hypokalaemic paralysis as initial manifestation of primary Sjögren’s syndrome is rare, but when it occurs it may precede symptoms of xerostomia and xerophthalmia. The diagnosis of primary Sjögren’s syndrome should be considered in young women who present with progressive weakness, hypokalaemia and metabolic acidosis.

The incidence of certain types of glomerular diseases is different in the elderly as compared with younger patients. Many different histologic appearances can be identified; however, membranous nephropathy is the most common type. Underlying malignancy has been thought to be responsible for 5 to 10 percent of cases of membranous nephropathy in adults, with the highest risk in patients over age 60. A solid tumor such as carcinoma of the lung or colon is most frequently involved. It is presumed that tumor antigens are being deposited in the glomeruli, which is followed by antibody deposition and complement activation, leading to epithelial cell and basement membrane injury and proteinuria due to the associated increase in glomerular permeability. In the present study, 33 patients aged over 60 with nephrotic syndrome were analyzed. Fourteen (42%) patients had membranous nephropathy, three of them with underlying malignancy., Incidencija pojedinih tipova glomerulonefritisa u starijih osoba razlikuje se u usporedbi s mlađim bolesnicima. U starijih osoba mogu se dijagnosticirati različiti oblici glomerulonefritisa, no najčešća je membranska nefropatija. Ovaj oblik glomerulonefritisa je u 5% do 10% bolesnika povezan s malignim tumorom. Najčešći oblici malignih tumora udruženih s membranskom nefropatijom su karcinomi pluća i kolona. Pretpostavlja se da se tumorski antigen odlaže u glomerulima, što izaziva taloženje protutijela i aktiviranje komplementa. To pak izaziva oštećenje epitelnih stanica i bazalne membrane glomerula te proteinuriju. U radu je analizirano 33 bolesnika s nefrotskim sindromom. Svi su bolesnici bili stariji od 60 godina. Membranska nefropatija je dijagnosticirana u 14 (42%) bolesnika, a kod troje bolesnika je ovaj oblik glomerulonefritisa bio udružen s malignom bolešću.

Renal side-effects of anti-inflammatory drugs (NSAID) can be diveded in five clinical syndromes: acute renal failure, acute intersticial nephritis with nephrotic syndrome, electrolyte and fluid disorders, hypertension and analgetic nephropathy.We described one unusual combination of acute tubular necrosis (ATN) and minimal change disease (MCD). Reports of ATN and MCD with nephrotic syndrome caused by NSAID exposure are very rare. This is the first reported case of MCD associated with ATN by Diclofenac. A 53-year-old woman with acute renal failure (creatinine was 716 μ mol/L) and nephrotic syndrome (she was in generalizied oedema and protein excretion was 6.0g/day) was admitted to our hospital. She also had eosinophilia. Her medical history was unremarkable except for nontreated hypertension for five years. Only medication she has been taking was Diclofenac for chronic muscular pain and knee arthropathy. NSAID drugs exert their toxic effect on the kidney by ihibition of prostaglandin synthesis and causing an acute allergic interstitial nephritis. The pathogenesis of NSAID-associated MCD is unclear. NSAID drugs cause inhibiton of cyclooxigenase and shift arachidonic acid toward lipooxygenase pathway, which may result in enhanced production of proinflammatory leukotriens. In addition, lypooxigenase products increase vascular permeability and may contribute by altering glomerular-cappilary barrier. The patient was treated with steroides. The response was excellent with remission of nephrotic syndrome and the normalization of renal excretory function within 6 weeks.