In vivo assessment of DNA damage induced in oral mucosa cells by fixed and removable metal prosthodontic appliances

Given long-term effect on oral tissues due to contact with dental appliances, the biocompatibility studies of casting alloys are of great importance. It has been previously documented that metal dental appliances, due to corrosion, might induce genotoxic and mutagenic effects in cells. Therefore, the aim of presented study was to examine the genotoxicity of two dental casting alloys (Co-Cr-Mo and Ni-Cr) commonly used in fixed and removable prosthodontic appliances that are in contact with the oral epithelium for 5 years or more. For that purpose, 55 age-matched subjects were included in the study; 30 wearers of prosthodontic appliances and 25 controls. Buccal cells of oral mucosa were collected and processed for further analysis. The cell viability has been assessed by trypan blue exclusion test, while genotoxic effect of metal ions on DNA in oral mucosa cells was studied by use of alkaline comet assay. Results have shown significantly higher comet assay parameters (tail length and percentage DNA in the tail) in the group wearing metal appliances. Both subjects with Co-Cr-Mo alloy and Ni-Cr alloy showed significantly higher comet assay parameters when compared with controls. It has been confirmed that metal ions released by the two base metal dental casting alloys examined in this study, might be responsible for DNA damage of oral mucosa cells. Therefore, the results of this study emphasize the importance of the in vivo evaluation of dental materials with respect to their genotoxicity, which is of major importance to ensure long-term biocompatibility.     

Morphometric analysis of renal arteries in patients with renal cell carcinoma

The aim of this study was to analyze morphometric parameters of renal arteries (longest diameter and tunica media thickness) in patients with renal cell carcinoma (RCC), to look into their relationship to tumor necrosis and to compare them with morphometric parameters recorded in a control group. We analyzed archival cases of RCC diagnosed in 2003 that also contained routinely sampled specimens of distal segments of renal artery. The control group consisted of specimens from both renal arteries obtained from 16 patients at routine autopsy during 2004-2005. Autopsy, as well as further histological analysis, did not disclose any malignant disease in the control group. Morphometric analysis of diameter and thickness of the renal artery tunica media was performed using Issa 3.1 software (Vamstek 2002, Zagreb, Croatia). The comparison of tunica media thickness showed that renal arteries from RCC cases were significantly thicker compared to distal parts of renal arteries in the control group (p=0.0002). Although renal artery samples from cases with necrotic tumor areas were thicker than those without tumor necrosis, the difference was not statistically significant. It is concluded that significantly thicker tunica media characterizes renal arteries in the group of patients with RCC when compared with the control group.     

Contribution of forensic anthropology to identification process in Croatia: examples of victims recovered in wells

Aim

To describe the contribution of forensic anthropology to the recovery, analysis, and identification of victims from the 1991-1995 war in Croatia recovered in wells.

Methods

From 1996 to the present, human remains of a total of 61 individuals have been recovered from 13 wells. Six wells contained the remains of a single individual, one well contained the remains of 2 individuals, and 6 wells contained the remains 3 or more individuals. The majority of wells, containing 90.2% (55/61) of recovered individuals, were located within a 4 km radius of the Croatian-Serbian border.

Results

Forensic anthropologists re-individualized 26/61 (42.6%) individuals out of skeletonized and commingled remains, provided basic biological data on sex, age-at-death, and stature in all identifications (n = 37), as well as established positive identification by recognizing unique skeletal features (antemortem fractures and skeletal evidence of antemortem surgical interventions) in 3/37 (8.1%) cases. Trauma analyses carried out by forensic anthropologists contributed to the determination of the cause of death in 38/61 (62.3%) individuals and to the probable cause of death in an additional 18/61 (29.5%) individuals. The most frequent (27/38, 71.0%) type of trauma causing death in individuals recovered from wells was a single gunshot wound.

Conclusion

Forensic anthropologists, collaborating closely with forensic pathologists, forensic odontologists, forensic radiologists, criminologists, and molecular biologists contributed significantly to trauma analysis and identification of war victims recovered from wells.To address the need for identification of a large number of war victims in the 1991-1995 war in Croatia (1-8), in 1991 the Croatian Government formed a multidisciplinary team of forensic experts with a purpose of recovering, analyzing, and identifying human remains from individual and mass graves. This report describes the contribution of forensic anthropologists to this endeavor, specifically to the recovery, analysis, and identification of 61 individuals recovered from 13 wells in Croatia after the 1991-1995 war. The recovery and analysis of these individuals was, for a number of reasons, considerably more complicated than the recovery and analysis of individuals from other types of mass graves and individual graves (1).For obvious reasons, the recovery of human remains deposited in wells was more hazardous than the recovery of remains from other types of settings. Standard risks associated with this type of work are difficult access to the grave sites through mine fields, possible explosive devices hidden among the remains, and poorly preserved human remains. Besides this, recovery from wells was complicated by varying depth of the wells, problems pertaining to the draining off the wells, and – in the case of more dilapidated wells – the risk of collapse of the well walls. Trauma analyses and identification of the recovered individuals was hampered by problems such as poor preservation and commingling of the remains which were significantly more present in wells than in other types of settings (1).The multidisciplinary team assembled by the Croatian government to recover and identify missing individuals included forensic pathologists, forensic anthropologists, forensic odontologists, forensic radiologists, criminologists, and molecular biologists. Forensic anthropologists contributed to this team effort by: 1) helping in locating and recovering human remains; 2) separating and re-individualizing commingled remains; 3) interpreting skeletal evidence of trauma – identifying the presence of trauma, differentiating between antemortem, postmortem, and perimortem trauma, as well as establishing the sequence of perimortem traumas, when possible; and 4) adopting an osteo-biographical approach in order to supply not only basic data on the age-at-death, sex, and stature, but also on other biological characteristics potentially useful in the identification process. To facilitate this, a joint US-Croatian forensic anthropology project was developed. In this project, forensic anthropologists from the Croatian Academy of Sciences and Arts and forensic specialists from the Department of Forensic Medicine at the School of Medicine, University of Zagreb, together with forensic anthropologists from the Smithsonian Institution, Washington DC, USA and the University of Tennessee in Knoxville developed a forensic anthropology database. Data on sex, age-at-death, stature, cranial and postcranial metric characteristics, and osteological and dental features were collected, including the evidence on pathology, ante-mortem fractures, disease, and surgical interventions, as well as perimortem trauma and possible cause of death. The purpose of this database was 3-fold: 1) to identify a basic and standard set of measurements, observations, and definitions to ensure that data are comparable; 2) to store the data in a computer so that particular subsets can be quickly accessed, analyzed, and compared; and 3) to provide up-to-date discriminant formulae for determining sex, stature, and other traits useful for comparative research and forensic analysis. The data gathered so far have already been proved useful for calculating discriminant functions for determining sex from fragmentary and complete Croatian femora (3), estimating male stature from the maximum lengths of the humerus, femur, and tibia (9), as well as determining craniometric relationships between Americans (Caucasians), Croatians, and Bosnians (10).     

Antithrombotics in heart failure

Heart failure is a common clinical condition associated with high morbidity and mortality rate despite significant improvements in pharmacotherapy and implementation of medical procedures. Patients with heart failure are at an increased risk of developing arterial and venous thrombosis, which contribute to the high rate of adverse events and fatal outcomes. Many heart failure patients routinely receive antithrombotic therapy due to the presence of a specific indication for its use, like ischemic heart disease or atrial fibrillation. However, there is no solid evidence to support the routine use of antithrombotic agents in all heart failure patients. This article reviews the evidence for using antithrombotic therapy in heart failure patients.Heart failure (HF) is a complex clinical syndrome caused by a number of different disorders that impair the heart muscle’s ability to maintain adequate blood flow to meet the body''s metabolic needs. Research and advances in managing HF so far have been mainly focused on improving the neurohormonal imbalance and assisting the myocardium to increase the cardiac output with different types of devices. Over the last couple of decades there have been considerable improvements in pharmacotherapy and medical procedures for treating HF, such as the introduction of angiotensin-converting enzyme inhibitors (ACEIs), angiotensin receptor blockers (ARBs), beta blockers, aldosterone antagonists, I_f channel blocker ivabradine, cardiac resynchronization therapy (CRT), and ventricular assist devices (VADs). However, HF is still a major public health issue associated with increased morbidity and mortality (1). Because of the broad spectrum of its clinical presentations, aging of the population, and different comorbidities and treatment options, HF treatment represents a considerable challenge. Patients with HF are at an increased risk of developing arterial and venous thrombosis, which contribute to the high rate of adverse events and fatal outcomes (2-5). Further research should clarify whether routine use of antithrombotic agents brings clinical benefit to all HF patients.     

New Drugs on the Block—Emerging Treatments for Nonalcoholic Steatohepatitis

Patients with nonalcoholic steatohepatitis (NASH) are at higher risk of progression to advanced stages of fibrosis, cirrhosis, hepatocellular carcinoma and othe...     

Radiological Impact of An Active Quarry in The Papuk Nature Park,Croatia

Papuk Nature Park, unlike most similar parks and preserves in the world, contains active quarries. Quarries dig stone from the ground, creating dust and exposing deeper, potentially more radioactive layers. Since the forest trails in the Park lead right up to the quarries, we believed it was important to determine the radiological impact of the quarries on the Park environment. We measured ambient dose rate equivalent H*(10) and sampled moss at 26 Park locations along two of four quarries, along the road between them, and near Lake Orahovac, a very popular tourist destination close to the quarries. Moss is a standard bioindicator of exposure to heavy metals, including radionuclides. Using-gamma ray spectrometry we determined the activity concentration of ¹³⁷Cs and of representative naturally occurring radionuclides – ²³⁸U, ²²⁶Ra, ²¹⁰Pb, ²³²Th, and ⁴⁰K – in sampled moss. H*(10) at selected locations was similar to the background H*(10) measured continuously all over Croatia. The ranges of measured activity concentrations of ¹³⁷Cs and naturally occurring radionuclides in moss did not differ significantly from other parts of Croatia and nearby countries.Key words: ¹³⁷Cs, ambient dose rate equivalent H*(10), gamma ray spectrometry, moss, naturally occurring radionuclides     

The lack of influence of IVS5-91 G>A polymorphism of the SCN1A gene on efficacy of lamotrigine in patients with focal epilepsy

ABSTRACT

Background: IVS5-91G>A (rs3812718) polymorphism of the sodium voltage-gated channel alpha subunit 1 (SCN1A) gene has been associated with inadequate responsiveness to common antiepileptic drugs which act as sodium channel blockers. This study was performed to investigate the effect of IVS5-91G>A (rs3812718) polymorphism on lamotrigine (LTG) efficacy in a cohort of patients with non-lesional focal epilepsy taking LTG as monotherapy.

Methods: A total of 100 of patients with non-lesional focal epilepsy on LTG monotherapy was included in this prospective interventional study. After reaching a stable dose of LTG patients were followed-up for 12 consecutive months. LTG responsiveness was defined as a 75% or more reduction in seizure frequency on a stable dose of LTG. Genotyping was performed at the end of the study using standard procedures and data were correlated with clinical data.

Results: There were no significant differences in the prevalence of responsiveness to LTG between carriers of different genotypes. Average maintenance LTG doses in the responder group differed by genotype in the order AA>GA>GG, but these differences did not reach statistical significance.

Conclusion: Our data suggest lack of association between SCN1A IVS5-91G>A (rs3812718) polymorphism and response to LTG.     

Influence of hyperthermal regimes on experimental teratoma development in vitro

We screened for the impact of hyperthermal regimes varying in the cumulative equivalent minutes at 43°C (CEM43°C) and media composition on tumour development using an original teratoma in vitro model. Rat embryos (three germ layers) were microsurgically isolated and cultivated at the air‐liquid interface. During a two week period, ectodermal, mesodermal and endodermal derivatives developed within trilaminar teratomas. Controls were grown at 37°C. Overall growth was measured, and teratoma survival and differentiation were histologically assessed. Cell proliferation was stereologically quantified by the volume density of Proliferating Cell Nuclear Antigen. Hyperthermia of 42°C, applied for 15 minutes after plating (CEM43°C 3.75 minutes), diminished cell proliferation (P ? .0001) and enhanced differentiation of both myotubes (P ? .01) and cylindrical epithelium (P ? .05). Hyperthermia of 43°C applied each day for 30 minutes during the first week (CEM43°C 210 minutes) impaired overall growth (P ? .01) and diminished cell proliferation (P ? .0001). Long‐term hyperthermia of 40.5°C applied for two weeks (CEM43°C 630 minutes) significantly impaired survival (P ? .005). Long‐term hyperthermia of 40.5°C applied from the second day when differentiation of tissues begins (CEM43°C 585 minutes) impaired survival (P ? .0001), overall growth (P ? .01) and cartilage differentiation (P ? .05). No teratomas survived extreme regimes: 43°C for 24 hours (CEM43°C 1440 minutes), hyperthermia in the scant serum‐free medium (CEM43°C 630 minutes) or treatment with an anti‐HSP70 antibody before long‐term hyperthermia 40.5°C from the second day (CEM43°C 585 minutes). This in vitro research provided novel insights into the impact of hyperthermia on the development of experimental teratomas from their undifferentiated sources and are thus of potential interest for future therapeutic strategies in corresponding in vivo models.