The Effect of Low-Intensity Pulsed Ultrasound on Bone Regeneration and the Expression of Osterix and Cyclooxygenase-2 during Critical-Size Bone Defect Repair.

Low-intensity pulsed ultrasound (LIPUS) is a form of ultrasound that utilizes low-intensity pulsed waves. Its effect on bones that heal by intramembranous ossification has not been sufficiently investigated. In this study, we examined LIPUS and the autologous bone, to determine their effect on the healing of the critical-size bone defect (CSBD) of the rat calvaria. The bone samples underwent histological, histomorphometric and immunohistochemical analyses. Both LIPUS and autologous bone promoted osteogenesis, leading to almost complete closure of the bone defect. On day 30, the bone volume was the highest in the autologous bone group (20.35%), followed by the LIPUS group (19.12%), and the lowest value was in the control group (5.11%). The autologous bone group exhibited the highest intensities of COX-2 (167.7 ± 1.1) and Osx (177.1 ± 0.9) expression on day 30. In the LIPUS group, the highest intensity of COX-2 expression was found on day 7 (169.7 ±1.6) and day 15 (92.7 ± 2.2), while the highest Osx expression was on day 7 (131.9 ± 0.9). In conclusion, this study suggests that LIPUS could represent a viable alternative to autologous bone grafts in repairing bone defects that are ossified by intramembranous ossification.

Trends in Professional Radiation Exposures of Medical Staff Covered by Personal Dose Monitoring at a Rijeka Clinical Hospital Centre (2000 to 2015).

ABSTRACT: The increase in the number of radiological procedures observed in recent years also means greater exposure to ionizing radiation for the medical staff performing these procedures. This is most pronounced in the teams that perform interventional and invasive radiological procedures. The aim of this study was to examine the impact of increasing numbers of radiological procedures on the effective dose received by medical staff and to determine which professions received the highest exposure to ionizing radiation. Data for effective doses of 326 employees of Rijeka Clinical Hospital Centre covered by personal dosimetry in the period from 2000 to 2015 have been analyzed. Employees were divided by sex, by departments in which they were employed, and according to their professions. The analysis has shown that the exposure level of workers working in the ionizing radiation zone is typically well below the dose limits. During 2015, most employees (over 96.3%) received an annual effective dose of less than 0.1 mSv. Only three persons received an annual dose higher than 0.5 mSv, and one person received an annual dose of 6.9 mSv. Comparison of the radiation exposure doses of medical workers of different professions has shown that the highest dose of radiation is received by cardiologists involved in interventional cardiology procedures. Therefore, the additional need is to take care of their protection, check the manner of their work, and ensure more even workload of cardiologists participating in procedures that involve higher exposure to ionizing radiation.

Matrix metalloproteinases MMP-1, MMP-2, and MMP-13 are overexpressed in primary nodular melanoma.

BACKGROUND: The spread and invasion of malignant melanoma cells involve degradation and reorganization of the extracellular matrix by the activation of several matrix metalloproteinases (MMPs). This study analyzed the expression of MMP-1, MMP-2, and MMP-13 proteins in primary nodular melanoma (NM) and dysplastic nevi (DN) as a significant risk factor for melanoma development. The secondary goal was to analyze the correlation of MMPs protein expression in NM with tumor invasion, BRAF V600 mutation status, and overall survival. METHODS: Immunohistochemistry for MMP-1, MMP-2, and MMP-13 was performed on nodular melanoma (n = 52) and dysplastic nevi (n = 28) on tissue microarray (TMA). BRAF V600 mutation analysis on NM samples was performed by the Sanger sequencing method. RESULTS: A high level of MMPs expression in NM samples (>30%) compared with DN (<8%) was statistically significant (P < 0.001). BRAF V600 mutations were detected in 15 of 39 (38.5%) NM samples. This study revealed an interesting finding that MMP-1 and MMP-13 protein expression in the BRAF V600 mutated melanomas were significantly lower than in the BRAF V600 wild type (P < 0.05). CONCLUSION: Cox analysis revealed that Clark categories, Breslow thickness, and MMP-1 high protein expression are predictive factors for shorter overall survival (P < 0.05).

Diagnostic value of tumour markers in pleural effusions.

INTRODUCTION: We investigated whether tumour markers carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), cancer antigen 125 (CA-125), and cytokeratin 19 fragment (CYFRA 21-1) in pleural effusions and serum can be used to distinguish pleural effusion aetiology. MATERIALS AND METHODS: During the first thoracentesis, we measured pleural fluid and serum tumour marker concentrations and calculated the pleural fluid/serum ratio for patients diagnosed with pleural effusion, using electrochemiluminescence immunoassays. Receiver operating characteristic (ROC) analysis was carried out and the Hanley and McNeil method was used to test the significance of the difference between the areas under ROC curves (AUCs). In order to detect which tumour marker best discriminates between malignant and non-malignant pleural effusions and to establish the predictive value of those markers, discriminant function analysis (DFA) and logistic regression analysis were utilized. RESULTS: Serum tumour markers CYFRA 21-1 and NSE as well as pleural NSE were good predictors of pleural effusion malignancy and their combined model was found statistically significant (Chi-square = 28.415, P < 0.001). Respective ROC analysis showed significant discrimination value of the combination of these three markers (AUC = 0.79). CONCLUSIONS: Serum markers showed superiority to pleural fluid markers in determining pleural fluid aetiology. Serum CYFRA 21-1 and NSE concentrations as well as pleural fluid NSE values had the highest clinical value in differentiating between malignant and non-malignant pleural effusions. The combination of these three markers produced a significant model to resolve pleural effusion aetiology.

Exploiting thermochromic materials for the rapid quality assurance of physiotherapy ultrasound treatment heads.

Significant nonuniformities in the acoustic intensity distribution generated by physiotherapy ultrasound treatment heads are not uncommon, potentially leading to significant localised temperature rises and tissue damage. An acoustic absorber tile containing a thermochromic pigment has been developed to provide rapid quality assurance of physiotherapy ultrasound treatment heads by virtue of a thermochromic colour change, indicating the time-averaged intensity distributions generated by these devices. As a bench-top device, the use of the tile is designed to mimic the nature of the physiotherapeutic application, requiring minimal training. Two designs where thermochromic pigments are added to the various polymeric layers of the tile are presented. Testing has been conducted with two physiotherapy treatment heads of differing performance, one of them notably exhibiting a strong "hot-spot" in localised acoustic time-averaged intensity. Findings show good qualitative agreement with classical hydrophone scans. Techniques are explored for the correction of nonlinearities in the thermochromic relationship, to enhance the accuracy of quantitative assessment.

Dosimetric verification of compensated beams using radiographic film.

INTRODUCTION: External photon beam modulation using compensators in order to achieve a desired dose distribution when brachytherapy treatment is followed by external beam radiation is a well-established technique. A compensator modulates the central part of the beam, and the dose beneath the thickest part of the compensator is delivered mostly by scattered, low energy photons. A two-dimensional detector with a good spatial resolution is needed for the verification of those beams. In this work, the influence of different types of detectors on the measured modulated dose distributions was examined. MATERIALS AND METHODS: Dosimetric verification was performed using X-Omat V, Eastman Kodak radiographic films at different depths in a solid water phantom. The film measurements were compared with those made by ionization chambers. Photon beams were also modelled using EGSnrc Monte Carlo algorithm to explain the measured results. RESULTS: Monte Carlo calculated over-response of the film under the thickest part of the compensator was over 15%, which was confirmed by measurements. The magnitude of over-response could be associated with changes in the spectra of photon energy in the beam. CONCLUSIONS: The radiographic film can be used for the dosimetry of compensated high energy photon beams, with limitations in volumes where photon spectra are hardly degraded.

Radiation therapy photon beams dose conformation according to dose distribution around intracavitary-applied brachytherapy sources.

Intracavitary application of brachytherapy sources followed by external beam radiation is essential for the local treatment of carcinoma of the cervix. Due to very high doses to the central portion of the target volume delivered by brachytherapy sources, this part of the target volume must be shielded while being irradiated by photon beams. Several shielding techniques are available, from rectangular block and standard cervix wedge to more precise, customized step wedge filters. Because the calculation of a step wedge filter's shape was usually based on effective attenuation coefficient, an approach that accounts, in a more precise way, for the scattered radiation, is suggested. The method was verified under simulated clinical conditions using film dosimetry. Measured data for various compensators were compared to the numerically determined sum of the dose distribution around brachytherapy sources and one of compensated beam. Improvements in total dose distribution are demonstrated, using our method. Agreement between calculation and measurements were within 3%. Sensitivity of the method on sources displacement during treatment has also been investigated.

Expression of Bcl-2 family proteins in psoriasis.

AIM: To elucidate the mechanisms involved in apoptosis of psoriatic keratinocytes by examining the expression of pro-apoptotic (Bak, Bax) and anti-apoptotic (Bcl-2, Bcl-X) Bcl-2 family of proteins, as well as the expression of p53 and Ki-67 proteins in normal skin, and uninvolved and involved psoriatic skin. METHODS: A total of 90 skin samples (30 cases of involved and uninvolved psoriatic skin and normal skin) were examined immunohistochemically to determine the protein expression of p53, Ki-67, Bcl-2, Bcl-X, Bax, and Bak. The results were quantified and expressed as a percentage of positive keratinocytes. RESULTS: There was a significant increase in Ki-67 (17.05 vs 3.65; P<0.001), Bcl-X (40.21 vs 13.97; P<0.001), Bak (89.46 vs 73.36; P<0.001), and Bax (50.00 vs 29.25; P<0.001) expression and a decrease in Bcl-2 (3.23 vs 6.25; P=0.008) expression in involved psoriatic skin, as well as an increase in Bcl-X (25.13 vs 13.97; P<0.001) expression in uninvolved psoriatic skin, when compared to normal skin. Samples with higher percentage of Ki-67 positive cells showed a higher percentage of p53 positive cells (correlation coefficient r=0.75 in involved psoriatic samples, P<0.001; r=0.88 in uninvolved psoriatic samples, P<0.001; and r=0.85 in normal skin samples, P<0.001). Samples with higher percentage of p53 positive cells expressed pro-apoptotic Bak and Bax in higher percentage of cells; the correlation coefficients were r=0.74 and r=0.68 in involved psoriatic samples (P<0.001 for both), r=0.75 and r=0.69 in uninvolved psoriatic samples (P<0.001, for both), and r=0.87 and r=0.70 in normal skin samples (P<0.001, for both). CONCLUSION: Increased expression of Bcl-X protein was associated with psoriatic epidermal hyperplasia. Strong Bax and Bak expression in involved psoriatic skin are probably inhibitory mechanisms counteracting intensive proliferation.

Expression of cell cycle and apoptosis regulatory proteins in keratoacanthoma and squamous cell carcinoma.

Some authors view keratoacanthoma (KA) as a variant of squamous cell carcinoma (SCC), while others consider it a separate entity that must be distinguished from SCC. Involution displayed by KA is an important difference between these two entities. It has been suggested that apoptosis plays a role in the involution process of KA, although the exact trigger for it remains unclear. A hundred and fifty specimens were included in this study, 30 cases for each of the following groups: normal skin (NS), proliferative keratoacanthoma (pKA), regressing keratoacanthoma (rKA), well-differentiated squamous cell carcinoma (wdSCC), and poorly differentiated squamous cell carcinoma (pdSCC). They were immunohistochemically examined for the expression of p53, Ki-67, bak, and bcl-2. Significantly higher p53 and Ki-67 expressions were observed in all tumor lesions examined as compared with NS. There was higher bak expression in KAs compared to NS and a significant reduction of bak expression in pdSCC together with a significant reduction of bak expression in SCCs compared to pKA. Bcl-2 expression was similar in NS and SCCs, but was lower in rKA. We found a significant positive correlation between p53 and Ki-67, p53 and Bak in NS and examined skin tumors. Lower bcl-2 expression in conjunction with higher bak expression in rKA suggests a possible role of these apoptosis-regulating proteins in tumor regression. In contrast to this finding, a steady level of bcl-2 expression in pdSCC combined with lower bak expression levels and a high proliferation rate could contribute to progression and aggressiveness in these tumors. Bak and p53 expression is a sun-related and age-dependent process in NS and skin tumors.

The role of perforin-mediated apoptosis in lichen planus lesions.

Lichen planus is recognized as a T-cell-mediated disease. Histologically, it is characterized by the formation of colloid bodies representing apoptotic keratinocytes. The apoptotic process mediated by CD8(+) cytotoxic T lymphocytes (CTLs) and NK cells mainly involves two distinct pathways: the perforin/granzyme pathway and the Fas/FasL pathway. So far, little is known regarding the role of perforin-mediated apoptosis in lichen planus. In the present study, the expression and distribution of perforin, T and NK cell subsets in the epidermis and dermis of lesional and nonlesional lichen planus skin were studied. Skin biopsy specimens from lesional and nonlesional skin of ten patients with lichen planus and eight healthy persons were analysed by immunohistochemistry. Significant accumulation of T cells, particularly of CD4(+) and CD8(+) subsets, was found in both epidermis and dermis of lichen planus lesions compared with nonlesional and healthy skin. There were no significant differences in the incidence of NK cells (CD16(+) and CD56(+)) between lesional, nonlesional and healthy skin. Perforin expression was significantly upregulated in the epidermis of lichen planus lesions. In conclusion, accumulation of perforin(+) cells in the epidermis of lichen planus lesions suggest a potential role of perforin in the apoptosis of basal keratinocytes.

Inflamed urachal cyst containing calculi in an adult.

The urachus is an embryonic structure that persists after birth in some individuals and can cause various problems. We report a case of an inflamed urachal cyst filled with a thick yellow fluid and several calculi in a woman with a 1-month history of dysuria. Physical examination revealed a fist-sized tumor located infraumbically in the midline. The patient's erythrocyte sedimentation rate was elevated; the results of all other routine laboratory studies were normal. Sonography showed a regularly shaped, ovoid, hypoechoic cystic area in the abdominal wall measuring 8 x 4 x 3 cm and containing several hyperechoic masses associated with acoustic shadowing. The wall of the cyst was inhomogeneous, and a thin hypoechoic linear tract linked the superior aspect of the mass to the umbilicus. The results of excretory urography, voiding cystography, and cystoscopy excluded an abnormality of the urinary system. A urachal cyst was diagnosed, and the mass was surgically removed. The surgical specimen was sent for histopathologic analysis, which confirmed the diagnosis.