Effect of SARS-CoV-2 antibody screening on participants' attitudes and behaviour: a study of industry workers in Split, Croatia.

OBJECTIVES: To investigate the changes in personal attitudes and behaviour before and after negative serological test results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies. STUDY DESIGN: Cross-sectional questionnaire survey. METHODS: A survey questionnaire was conducted with 200 industry workers (68% males and 32% females) who had previously tested negative for SARS-CoV-2 antibodies. The survey examined participants' self-reported general attitudes towards coronavirus disease 2019 (COVID-19), their sense of fear, as well as their behaviour towards protective measures before and after the testing. RESULTS: Participants perceived the disease as a severe health threat and acknowledged that the protective measures were appropriate. Respondents reported a high level of adherence to measures and low level of fear, both before and after the testing. Although these indicators were statistically significantly reduced after the test (P < 0.004), they did not result in irresponsible non-adherence behaviours. Almost all respondents attributed their application of personal protection measures to factors other than the results of serological screening. CONCLUSIONS: Serological tests do not contribute to irresponsible non-adherence behaviours in an environment where protective measures are efficient. However, they may help reduce fear within society and working environments.

Expression of Ki-67 and p53 in meningiomas.

UNLABELLED: Meningiomas account for about 30% of all primary brain tumors. It is difficult to predict the behaviour of meningiomas, and identification of protein markers responsible for the regulation of cell proliferation can be very helpful. The aim of this study was to evaluate immunohistochemical expression of Ki-67 and p53 in 170 meningiomas.A total number of 170 meningioma samples were classified according to WHO, immunohistochemically stained for Ki-67 and p53 and analysed using light microscope. Of 170 meningiomas analysed, 142 were grade I, 17 grade II and 11 grade III. Female to male ratio was 1.42:1. Statistically significant correlation was found between tumor grade and Ki-67 (p<0.001). There was significant correlation between Ki-67 levels and tumor subtypes (p=0.009). The optimal cut-off value for Ki-67 was 3.195. Tumors with Ki-67 ≤3.195 were 2 cm smaller than tumors with Ki-67 >3.195. Statistically significant correlation was found regarding p53 expression and tumor size (p=0.034). No correlation was established between Ki-67 or p53 and location of the tumor.According to positive correlation between tumor grade and subtype with Ki-67 levels, as well as positive correlation between Ki-67 and p53 with tumor size, indicate that Ki-67 and p53 might have influence on meningioma development and progression. KEYWORDS: meningioma, Ki-67, p53, immunohistochemistry.

Anterior ischemic optic neuropathy comorbid with Factor V Leiden and PAI-1 4G/5G mutation.

Acute anterior ischemic optic neuropathy (AION) is a disabling disease which impairs visual functions. AION is characterized by ischemic injury of the optic nerve caused by hypoperfusion in the short posterior ciliary arteries supplying the optic nerve head. AION is a complex multi-factorial disease and is difficult to diagnose based on clinical symptoms and signs alone. We describe the case of a 54-year-old patient affected by AION, with only known risk factors were PAI-1 4G/5G and Factor V Leiden mutation with hyperlipidaemia (Ref. 36). Full Text (Free, PDF) www.bmj.sk.

DNA typing from skeletal remains: evaluation of multiplex and megaplex STR systems on DNA isolated from bone and teeth samples.

AIM: To evaluate the performance of three multiplex short tandem repeat (STR) systems (AmpflSTR Profiler, AmpflSTR Profiler Plus, and AmpflSTR COfiler), and a megaplex STR system (PowerPlex 16) on DNA extracted from the skeletal remains. By performing a microbial DNA challenge study, we also evaluated the influence of microbial DNA on human DNA typing. METHODS: A subset of 86 DNA extracts isolated from 8-50 years old bone and teeth samples, corresponding to 20 identification cases from mass graves in Croatia and Bosnia and Herzegovina, and to 4 paternity cases involving deceased parents in Spain, were analyzed by the above systems. RESULTS: Bone samples with no detectable human DNA (tested with Quantiblot), as well as teeth samples with detectable human DNA, were successfully amplified. Surprisingly, even in highly degraded samples, PowerPlex 16 offered very robust amplification for the both Penta E and Penta D markers. We observed a few non-specific extra peaks of 202 and 308 base pairs, which appeared to match 16S rRNA of the Pseudomonas halodenitrificans. CONCLUSION: AmpflSTR Profiler Kit, AmpflSTR Profiler Plus Kit, the AmpflSTR COfiler Kit, and the PowerPlex 16 system are very sensitive multiplex STR amplification systems, which can be successfully used to obtain a multilocus STR profile from old teeth and bone samples with minimal amounts (pg) of human DNA or even with no detectable human DNA.

Population variation of human mitochondrial DNA hypervariable regions I and II in 105 Croatian individuals demonstrated by immobilized sequence-specific oligonucleotide probe analysis.

AIM: To detect sequence variation in 105 Croatian individuals by the use of duplex polymerase chain reaction amplification of full-length hypervariable region I and II (HVI/HVII) products and subsequent hybridization to a linear array of 27 immobilized sequence-specific oligonucleotide (SSO) probes, which targets six regions within HVI and HVII, and two additional sites, 189 and 16093. METHODS: Chelex-extracted bloodstains were used for amplification of HV regions. In all cases, a single robust amplification was sufficient for immobilized SSO probe typing and subsequent direct sequence analysis for both HVI and HVII. This method, suitable for a range of forensic samples (including shaft portions of single hairs), was also applied to the analysis of 18 skeletal elements recovered from a mass grave. Using a panel of immobilized SSO probes, we have developed a rapid screening approach to mitochondrial DNA (mtDNA) haplotyping before direct sequence analysis. RESULTS: We established a reference sequence database of mtDNA haplotypes for 105 randomly selected Croatian individuals. Fifty different mitotypes were observed (33 unique). The most frequent mitotypes occurred 18 times or approximately 17.1% [111111 189 (A) 16093 (T)] and 11 times or approximately 10.5% [131111 189 (A) 16093 (T)]; all other mitotypes occurred 5% or less. The corresponding genetic diversity value for this database was approximately 0.952. The usefulness of establishing an mtDNA reference database with immobilized SSO probe testing has been demonstrated by determining the strength of a match comparison obtained for one skeletal element and a corresponding maternal reference from 18 specimens recovered from a mass grave. CONCLUSION: The sequence variation detected by the panel of immobilized SSO probes is sufficiently diverse to be used for identification of human skeletal remains from mass graves. The immobilized SSO typing strip targets polymorphic regions within HVI and HVII and is a useful identification tool for mass grave and mass disaster analysis, as well as for criminal casework testing.

Expression of erbB-3 protein in colorectal adenocarcinoma: correlation with poor survival.

BACKGROUND/AIMS: The family of erbB receptors includes four transmembrane glycoproteins with tyrosine kinase activity. These receptors are widely expressed in normal tissues, but they also have been implicated in the development of several human adenocarcinomas. c-erbB-3/HER-3 has been detected to a greater or lesser extent in many tissues from the digestive, urinary, reproductive and respiratory tracts. The overexpression of c-erbB-3/HER-3 protein has also been shown in 53%-88% of colorectal adenocarcinomas. In this study we investigated the expression of the c-erbB-3/ HER-3 gene product in colorectal tumour samples, and compared the results obtained with several clinicopathological parameters, including the survival of patients. METHODS: Paraffin-embedded tissue sections were analysed immunohistochemically, using monoclonal antibody RTJ1 to human erbB-3 protein. Antibody RTJ1 specificity was confirmed by immunoprecipitation followed by Western blotting analysis. Amplification of the erbB-3 oncogene was tested by dot-blot hybridization. RESULTS: Adenocarcinomas of the colon were positive for erbB-3 protein in 78% of samples examined. Dot-blot analysis showed no amplification of the erbB-3 gene in colon adenocarcinomas. Statistical analysis showed that patients with tumours that could not be stained for erbB-3 protein survived significantly longer (P<0.05) than patients with tumours staining positive for the erbB-3 protein. A Cox proportional-hazards model with stepwise variable selection identified age, sex and erbB-3 expression as important prognostic factors. CONCLUSION: These findings demonstrate that erbB-3 protein expression could serve as a prognostic factor in colorectal malignancies.

[Incidence of stroke in central Dalmatia during the war in the Republic of Croatia (1991-1995)]. / Incidencia de ictus en la región de la Dalmacia media durante la guerra en la República de Croacia (1991-1995).

INTRODUCTION AND METHOD: A retrospective study was made of the incidence of stroke in the Dalmatian region (Split) during the war in the Republic of Croatia (1991-1995), to determine whether the war had affected the incidence of stroke in the population at risk. The data obtained was compared with data regarding the incidence of strokes in the pre-war period (1986-1990). RESULTS AND CONCLUSIONS: There was a small but significant increase in the incidence of strokes during the war period. There was also a considerable increase in patients with primary intracerebral hemorrhage during this period. The possible causes of this increase in patients with primary intracerebral hemorrhage are discussed.

Peripheral nerve war injuries.

OBJECTIVE: The purpose of this study is to evaluate peripheral nerve war injuries sustained during the war in southern Croatia and Bosnia and Herzegovina. PATIENTS AND METHODS: During the war in Croatia, 713 patients (99% male and 1% female) with wounds inflicted by firearms were examined at the Laboratory of Neurophysiology, University Hospital, Split. The patients, soldiers and civilians alike, ranged in age from 6 to 73 years (average, 28 years). All patients with firearm nerve war injuries underwent detection by electromyography and plurisegmental examination of the damaged peripheral nerve (neurography). The patients were examined and controlled on three occasions: within 2 months after wounding; up to 6 months after wounding; and more than 6 months after wounding. RESULTS: Single peripheral nerve lesions were present in 80% of the patients, and multiple peripheral nerve or plexus lesions were present in 20% of the patients. Peroneal and ulnar nerves were most often involved (20.9% and 19.8%, respectively). Associated massive injuries to the muscles, large blood vessels, or vital organs were present in 45% of the patients. Wounds were inflicted by shell fragments in 80% of the patients and by projectiles in 20% of the patients. CONCLUSION: According to our results, better recovery was achieved with conservative treatment and when physical therapy was initiated early with maximal patient cooperation. Electromyoneurographic findings were the most valid in the prognostic classification of war-inflicted peripheral nerve injuries.

Genetic analysis of the glucose-6-phosphate dehydrogenase deficiency in a southern Croatia.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. G6PD Mediterranean is caused by a C-->T transition at nucleotide 563, is characterized with less than 10% of normal enzyme activity and is classified as severe G6PD deficiency. Nineteen unrelated males from Southern Croatia with severe G6PD deficiency were tested, by enzyme digestion, for the presence of the Mediterranean mutation. Individuals with G6PD Mediterranean were further screened for the silent C-->T transition at nucleotide 1311. Four of the nineteen individuals were positive for the Mediterranean mutation (21%) and all four had the silent mutation.

Two new cases of liver angiosarcoma: history and perspectives of liver angiosarcoma among plastic industry workers.

In this report of two new cases of liver angiosarcoma (ASL) among plastic industry workers, the authors present the history and perspectives of this problem. The first cases of ASL have been registered since 1974, and in 1984, the European register of angiosarcoma was founded. In this register, 11 cases of ASL and one case of haemangiopericytoma have been registered from Croatia, all from a single plastics plant near Split. Two new cases of ASL (in retired autoclave cleaners, who were exposed to a concentration of 500-1000 ppm vinyl chloride monomer (VCM) during the working process) in the same plant are represented. They were detected with combined techniques of ASL detection, and both are still alive. The diagnoses have been histologically confirmed: one of them was surgically treated with segmental liver resection. The appearance of new cases of ASL confirms the perspective presented in the last report by the same authors.

The expression of p185(HER-2/neu) correlates with the stage of disease and survival in colorectal cancer.

BACKGROUND & AIMS: HER-2/neu oncogene encodes a transmembrane tyrosine kinase receptor that is amplified and/or overexpressed predominantly in adenocarcinomas. This phenomenon has been most intensively studied in breast carcinoma where its amplification and overexpression correlate with the overall course of disease and poor prognosis. This study was designed to investigate HER-2/neu gene expression in benign and malignant colorectal lesions and to evaluate its prognostic importance in colorectal cancer. METHODS: Two hundred twenty-one samples of normal colon, benign lesions, and colorectal adenocarcinomas were studied for expression of HER-2/neu oncoprotein. Immunohistochemical staining of formalin-fixed, paraffin-embedded tissue sections of primary tumor and lymph nodes was performed. Immunoprecipitation followed by Western blotting of freshly frozen samples of the same tumors were also performed. RESULTS: Normal colon mucosa, benign lesions, and adenocarcinomas clearly differed in the expression levels and histological distribution of p185(HER-2/neu). Normal mucosa was mostly negative, but significant number of benign lesions and adenocarcinomas overexpressed HER-2/neu protein. Adenocarcinomas were significantly more positive than benign lesions. The results show significant correlation with the epithelial abnormality degree and clinical parameters including Dukes' classification and relapse-free and postoperative survival period. CONCLUSIONS: The p185(HER-2/neu) rate expression could serve as an independent prognostic factor in patients with p185(HER-2/neu)-positive colorectal malignancies.

Nucleolar organizer region in transitional cell carcinoma of the bladder.

The silver staining technique that identifies NORs (nucleolar organizer regions) associated proteins was used for examining changes in nucleolar organizer region numbers in transitional cell carcinoma of the bladder. This method reveals NORs as black dots in the cell nuclei, and the number of NORs per cell (NORs/cell) has been taught to be related to cellular activation and to be a possible predictor of clinical outcome. A hundred specimens of transitional cell carcinoma of the bladder divided into four histologic grades (Ash classification) with 25 specimens each, and 25 specimens of normal bladder mucosa were analyzed. It has been demonstrated that the amount of protein synthesized by carcinoma varies according to its histologic grade, i.e. the higher histologic grade, the greater the NORs/cell number for each histologic grade. The characteristic mean NORs/cell SD was determined (GI 4.85 +/- 0.82, GII 5.94 +/- 1.42, GIII 8.54 +/- 1.01 and GIV 8.72 +/- 1.42), among which statistically significant differences were found. For similar histologic grades the characteristic mean NORs/cell +/- SD showed no statistically sex and age related differences. In some cases the mean NORs/cell for the examined carcinoma did not match the characteristic mean NPRs/cell +/- SD for the respective histologic grade.

Early intracranial infections after brain missile injuries--the role of computer tomography in diagnosis and treatment.

During the war in Croatia, from August 1991 until December 1994, 138 soldiers were treated at the Split University Hospital for different brain injuries inflicted by missiles. Nine of these 138 patients developed intracranial infection. This retrospective study reports on the outcome of these 9 intracranial infections caused by penetrating missile head injuries. In case of clinically suspected infection, computerized tomography scan was obtained at two time points during the course of infection in the same patient. Scans were obtained with and without contrast media, 7 to 14 days after the injury and the 4 weeks later. The role of computerized tomography in the detection and follow-up of various intracranial infections and long-term consequences were evaluated.

Ciliated metaplasia in a patient of Mediterranean origin with gastric adenoma.

We report here the first case of ciliated gastric metaplasia in a Croatian patient. This is also the first case of ciliated metaplasia reported in a patient of Mediterranean descent. Cilia were found in slightly cystically dilated gastric glands underneath a gastric adenoma with severe dysplasia. They were visualized by desmin immunohistochemical stain. Cells that presented with cilia were columnar cells, some of them with vacuolization of the cytoplasm. This case report shows that ciliated metaplasia occurs in patients of Southern European origin.

Identification of war victims from mass graves in Croatia, Bosnia, and Herzegovina by use of standard forensic methods and DNA typing.

The postmortem remains of sixty-one war victims were excavated from 6 mass graves in Bosnia and Herzegovina one and a half years after interment Using standard identification methods, including the matching of medical and dental records, the recognition of distinguishing characteristics such as the use of clothing and belongings, and video superimposition, 35 persons were identified. For the remaining 26 persons identification efforts continue. DNA typing was performed at the HLA DQA1 locus and five PM system loci. Results from DNA typing were confirmed by other methods. DNA profiles of family members of 150 missing persons are now being developed using the 6 loci. These DNA profiles will then be compared with those generated from the bone and teeth remains of the unidentified victims.

Northern and southern Croatian population data on seven PCR-based loci.

Northern and southern Croatian sample populations were typed at seven PCR-based loci -LDLR, GYPA, HBGG, D7S8, Gc, HLA-DQA1 and D1S80. The results show that all loci meet Hardy-Weinberg expectations and that there is little evidence for association of alleles between loci. Allelic frequency distributions at all loci, except HLA-DQA1, show no differences between the northern and southern Croatian sample populations. Moreover, the population data for Croatians are similar to U.S. Caucasians; only HLA-DQA1 for southern Croatians was statistically different compared with U.S. Caucasians. A Croatian population database(s) has been created and can be used for forensic analyses to estimate the frequency of a multiple locus DNA profile.

Vinylchloride monomer exposure by the plastic industry workers basic condition for liver angiosarcoma appearance.

The retrospective study of vinylchloridemonomer (VCM) exposure at 37 workers in the period from 1969 till 1987 was done, average time of exposition was 1.843 +/- 1.105 h, average VCM concentration was 543 +/- 228 ppm and average total exposure amount ("cumulative concentration of VCM") was 1.103811 +/- 1.050 766 ppmh at the time of autoclave cleaning 0.4 +/- 0.1 h. According updata investigations, high VCM concentration is essential for liver angiosarcoma (ASL) occurrence although the length of exposure is of concern. The possibility of new cases of angiosarcoma of liver is warned about.