Studies on healthy and neoplastic tissues using positron annihilation lifetime spectroscopy and focused histopathological imaging.

Samples of healthy and neoplastic myometrial tissues were investigated using Positron Annihilation Lifetime Spectroscopy (PALS). Meaningful differences between normal and diseased tissues were observed for each patient. The differences were also clearly visible for various kinds of lesions in each patient. The set of lifetimes and intensities obtained from PALS was correlated with the histopathological examinations of the same fragments of tissues. Strong coincidence between PALS parameters and histopathological findings was observed only in the case of a very precise correlation of the investigated area in both techniques. Measurements and discussion presented here were carried out to develop a method for measuring the sub-nanometric structure of human tissues. This kind of investigation, using positron probe, creates an opportunity of a new application in Positron Emission Tomography (PET).

Evaluation of ALK gene rearrangement in central nervous system metastases of non-small-cell lung cancer using two-step RT-PCR technique

Purpose. RT-PCR technique has showed a promising value as pre-screening method for detection of mRNA containing abnormal ALK sequences, but its sensitivity and specificity is still discussable. Previously, we determined the incidence of ALK rearrangement in CNS metastases of NSCLC using IHC and FISH methods. Materials. We evaluated ALK gene rearrangement using two-step RT-PCR method with EML4-ALK Fusion Gene Detection Kit (Entrogen, USA). The studied group included 145 patients (45 females, 100 males) with CNS metastases of NSCLC and was heterogeneous in terms of histology and smoking status. Results. 21% of CNS metastases of NSCLC (30/145) showed presence of mRNA containing abnormal ALK sequences. FISH and IHC tests confirmed the presence of ALK gene rearrangement and expression of ALK abnormal protein in seven patients with positive result of RT-PCR analysis (4.8% of all patients, 20% of RT-PCR positive patients). RT-PCR method compared to FISH analysis achieved 100% of sensitivity and only 82.7% of specificity. IHC method compared to FISH method indicated 100% of sensitivity and 97.8% of specificity. In comparison to IHC, RT-PCR showed identical sensitivity with high number of false positive results. Conclusion. Utility of RT-PCR technique in screening of ALK abnormalities and in qualification patients for molecularly targeted therapies needs further validation (AU)

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Evaluation of ALK gene rearrangement in central nervous system metastases of non-small-cell lung cancer using two-step RT-PCR technique.

PURPOSE: RT-PCR technique has showed a promising value as pre-screening method for detection of mRNA containing abnormal ALK sequences, but its sensitivity and specificity is still discussable. Previously, we determined the incidence of ALK rearrangement in CNS metastases of NSCLC using IHC and FISH methods. MATERIALS: We evaluated ALK gene rearrangement using two-step RT-PCR method with EML4-ALK Fusion Gene Detection Kit (Entrogen, USA). The studied group included 145 patients (45 females, 100 males) with CNS metastases of NSCLC and was heterogeneous in terms of histology and smoking status. RESULTS: 21% of CNS metastases of NSCLC (30/145) showed presence of mRNA containing abnormal ALK sequences. FISH and IHC tests confirmed the presence of ALK gene rearrangement and expression of ALK abnormal protein in seven patients with positive result of RT-PCR analysis (4.8% of all patients, 20% of RT-PCR positive patients). RT-PCR method compared to FISH analysis achieved 100% of sensitivity and only 82.7% of specificity. IHC method compared to FISH method indicated 100% of sensitivity and 97.8% of specificity. In comparison to IHC, RT-PCR showed identical sensitivity with high number of false positive results. CONCLUSION: Utility of RT-PCR technique in screening of ALK abnormalities and in qualification patients for molecularly targeted therapies needs further validation.

Sensitive methods for screening of the MEK1 gene mutations in patients with central nervous system metastases of non-small cell lung cancer

Background: The mitogen-activated protein kinases 1 and 2 (MEK1, MEK2) are fundamental partners in the RAS-RAF-MEK-ERK pathway that is involved in regulation of cell proliferation, differentiation and survival. Downregulation of the MEK cascades has been implicated in acquiring of the malignant phenotype in various cancers. Somatic mutations in MEK1 gene (substitutions K57N, Q56P, D67N) were described in < 1 % of non-small cell lung cancer (NSCLC) and they were more commonly reported in adenocarcinoma patients with current or former smoking status. Materials and methods: In the following study, we assessed the MEK1 gene mutations in 145 FFPE tissue samples from central nervous system (CNS) metastases of NSCLC using HRM-PCR and ASP-qPCR techniques. The studied group was heterogeneous in terms of histopathology and smoking status. The prevalence of the MEK1 gene mutation was correlated with the occurrence of mutations in KRAS, EGFR, DDR2, PIK3CA, NRAS, HER2, AKT1 and PTEN genes. Results: Using HRM and ASP-qPCR methods we identified one (0.7 %; 1/145) MEK1 substitution (Q56P) in CNS metastases of NSCLC. The mutation was identified in a single, 50-year-old, current smoking men with adenocarcinoma (1.25 %; 1/80 of all adenocarcinomas). Conclusions: According to the current knowledge, the incidence of MEK1 gene mutation in CNS metastatic lesion of NSCLC is the first such report worldwide. The analysis of gene profile in cancer patients may extend the scope of molecularly targeted therapies used both in patients with primary and metastatic tumors of NSCLC (AU)

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Sensitive methods for screening of the MEK1 gene mutations in patients with central nervous system metastases of non-small cell lung cancer.

BACKGROUND: The mitogen-activated protein kinases 1 and 2 (MEK1, MEK2) are fundamental partners in the RAS-RAF-MEK-ERK pathway that is involved in regulation of cell proliferation, differentiation and survival. Downregulation of the MEK cascades has been implicated in acquiring of the malignant phenotype in various cancers. Somatic mutations in MEK1 gene (substitutions K57N, Q56P, D67N) were described in <1 % of non-small cell lung cancer (NSCLC) and they were more commonly reported in adenocarcinoma patients with current or former smoking status. MATERIALS AND METHODS: In the following study, we assessed the MEK1 gene mutations in 145 FFPE tissue samples from central nervous system (CNS) metastases of NSCLC using HRM-PCR and ASP-qPCR techniques. The studied group was heterogeneous in terms of histopathology and smoking status. The prevalence of the MEK1 gene mutation was correlated with the occurrence of mutations in KRAS, EGFR, DDR2, PIK3CA, NRAS, HER2, AKT1 and PTEN genes. RESULTS: Using HRM and ASP-qPCR methods we identified one (0.7 %; 1/145) MEK1 substitution (Q56P) in CNS metastases of NSCLC. The mutation was identified in a single, 50-year-old, current smoking men with adenocarcinoma (1.25 %; 1/80 of all adenocarcinomas). CONCLUSIONS: According to the current knowledge, the incidence of MEK1 gene mutation in CNS metastatic lesion of NSCLC is the first such report worldwide. The analysis of gene profile in cancer patients may extend the scope of molecularly targeted therapies used both in patients with primary and metastatic tumors of NSCLC.

Subdural infusion of dexamethasone inhibits leukomyelitis after acute spinal cord injury in a rat model.

Trauma in spinal cord injury often results in massive damage to the white matter and in damage to myelin that results in a severe phagocyte-rich infiltration apparently directed at removing immunologically toxic myelin debris. In the epidural balloon crush injury to the rat cranial thoracic spinal cord, the dorsal column was crushed, which at one week post-op resulted in its obliteration by a severe infiltration by a virtually pure population of macrophages that internalized all damaged myelin. A week-long subdural infusion of dexamethasone, a stable synthetic corticosteroid, resulted in remarkable inhibition of the macrophage infiltration of the crush cavity and in the lack of removal of myelin debris by phagocytosis. In this study we demonstrated that spinal cord injury results in a severe inflammatory response directed at massively damaged myelin, and we inhibited this response with a subdural infusion of a powerful anti-inflammatory drug, dexamethasone.

Novel dipyrazolopyridine derivatives as deep blue emitters for polymer based organic light emitting diodes.

Paper reports synthesis and spectroscopic properties of four newly synthesized dipyrazolo[3,4-b; 3',4'-e]pyridine (DPP) derivatives. The spectroscopic studies are supplemented by quantum-chemical calculations using DFT/TDDFT/PCM method at the B3LYP/6-31+G(d,p) level of theory. The optical absorption and fluorescence emission processes appear to be weakly dependent on attached side phenyl and/or methyl groups exhibiting in cyclohexane solution the first absorption band (00' transition) in the region of 386-401 nm and the fluorescence band (0'0 transition) in the range of 412-425 nm. The electroluminescence devices (OLEDs) with an active PVK layer doped by DPP dyes have been designed. All the devices exhibit deep blue electroluminescence with the emission maximum being rather weakly dependent on the type of the fluorescent dopant. The obtained results demonstrate that a series of newly synthesized DPP dyes may be considered as perspective blue fluorescent emitters for electroluminescent applications.

Photoluminescence of 1-phenyl,3-methyl pyrazoloquinoline derivatives.

Paper presents the absorption and photoluminescence of 7-TFM, 6-F, 6-CN, 6-TBu and 6-COOEt derivatives of 1-phenyl-3-methyl-1H-Pyrazolo[3,4-b]quinoline (MPPQ). The measured spectra are compared with the results of the quantum chemical calculations performed by means of the semi-empirical methods (AM1 or PM3) that have been applied either to the equilibrium molecular conformations in vacuo (T=0K) or combined with the MD simulations (T=300 K). The photoluminescent spectra of MPPQ dyes are highly solvatochromic. The emission bands broaden and shift to the red with the increasing of solvent polarity, indicating thus a substantial dipole moment of the excited states. According to the quantum chemical analysis the reason for the strong solvatochromism of MPPQ dyes is related with intramolecular charge transfer (ICT) state. Due to the large dipole moment in the twisted geometry the ICT state is believed to become the lowest excited state in a strongly polar environment. This would explain a considerable solvatochromic shift in the highly polar solvents observed for all MPPQ dyes in the experiment. Such hypothesis is supported by the semi-empirical quantum chemical evaluations.

Optical absorption of methoxy and carboethoxy derivatives of 1,3-diphenyl-1H-pyrazolo[3,4-b]quinoline.

Paper deals with experimental investigations and quantum chemical calculations of the optical absorption spectra of methoxy and carboethoxy 1,3-diphenyl derivatives of the pyrazoloquinoline ([PQ]): 6-methoxy-1,3-dyphenil-[PQ], 6-methoxy-1,3-(p-methoxyphenyl)-[PQ], 6-methoxy-1-(p-methoxyphenyl)-[PQ] and 6-carboethoxy-1,3-diphenyl-[PQ]. The quantum chemical calculations are performed by means of the semiempirical quantum chemical methods (AM1 or PM3) applied to: (a) the equilibrium molecular conformation in vacuo (T=0 K); (b) the molecular dynamic (MD) trajectory (T=300 K) which includes the dynamics of a certain molecular fragment (moiety) only (fragmental MD simulations); or (c) the MD trajectory obtained for most general case within the total MD simulations at T=300 K. The results of these calculations are compared with the measured spectra of the optical absorption. The quantum chemical simulations show that the dynamics of the methoxy or carboethoxy groups practically does not influence the absorption spectrum whereas the strongest its modification (300

Optical poling effect and optical absorption of cyan, ethylcarboxyl and tert-buthyl derivatives of 1H-pyrazolo[3,4-b]quinoline: experiment and quantum-chemical simulations.

We present here results of experimental studies and quantum-chemical simulations of optical absorption and optical poling effects performed on a new synthesized cyan, ethylcarboxyl and tert-buthyl derivatives of 1H-pyrazolo[3,4-b]quinoline incorporated into polymer matrix or dissolved in organic solutions. The efficiency of second-order optical susceptibility d vs photoinduced power density I(p) clearly saturates to certain magnitude d(eff) at sufficient power densities (I(p) > or = 1.3 GW cm(-2)). Comparing experimental data and results of semiempirical quantum-chemical simulations one can conclude that there exists generally a good correlation between the magnitude of saturated susceptibilities d(eff) and macroscopic hyperpolarizabilities for all compounds except the chromophore 1,3-dimethyl-6-cyano-[PQ] only. The discrepancy for this compound may reflect a specific contribution of surrounding polymer matrix. According to the quantum chemical analysis the methyl-containing cyan and ethylocarcoxyl derivatives reveal four/five strong absorption bands in the spectral range 200-500 nm. A substitution of the methyl groups by the phenyl group causes the substantial changes of the absorption spectra mainly in the spectral range 240-370 nm. Measured and calculated absorption spectra manifest rather good agreement mainly in the part regarding the spectral positions of the first oscillator (absorption threshold). The quantum-chemical PM3 method shows the best agreement with experiment. At the same time a considerable broadening almost of all absorption bands appears as a characteristic feature of all measured spectra. The discrepancies between the calculated and the measured spectra are attributed to electron-vibronic coupling as well as to a specific rotational dynamics of phenyl rings.

[Expression of bcl-2 gene in glia-derived tumors of the brain]. / Ekspresja genu bcl-2 w glejopochodnych guzach mózgu.

Bcl-2 gene is an important factor in regulation of apoptosis as an inhibitor of this process. Its increased expression is observed in numerous neoplasms, what points out on importance of this gene in the process of oncogenesis. The aim of this paper was to assess the expression of bcl-2 gene in the cells of the glia-derived tumours of the brain. The method of Chomczynski and Sacchi with guanidine tiocyanate was applied to isolate a whole cellular RNA. Expression of bcl-2 gene was measured with a reverse transcription method by a synthesis of cDNA and amplification of gene fragment with specific oligonucleotides in polymerase chain reaction (RT-PCR). The expression of bcl-2 gene was found in cells of all examined glial tumours at the level of transcript. We carried out the discussion of the results and an attempt to explain the importance of bcl-2 gene expression of these neoplasms.

Online Internet-based robotic telepathology in the diagnosis of neuro-oncology cases: a teleneuropathology feasibility study.

This feasibility study examined the diagnostic accuracy of Internet-based dynamic-robotic telepathology using neuropathology cases. Randomly, 83 cases were selected from the routine diagnostic workload of the Neurosurgical Pathology Laboratory in Poznan, Poland. Telepathology diagnoses were compared with conventional paraffin section diagnosis. The neuropathologists, operating a robotically controlled motorized microscope over the Internet from 3 different Polish cities, individually reviewed the cases using computer workstations. Viewing times ranged from 2 minutes 54 seconds to 32 minutes 12 seconds per case. The mean diagnostic accuracy for telepathology diagnosis was 95%, with 2 of 3 observers achieving 100% diagnostic accuracy. Image quality was judged to be sufficient for correct evaluation, and the viewing times required to establish a final diagnosis by remote video microscopy were acceptable. Generally, user acceptance of robotic telepathology was high.

3-D temperature distribution in ultrasound hyperthermia with interstitial waveguide applicator.

Knowledge of temperature distribution in thermal treatment of cancerous tissue is of primary importance for the therapy success. We discuss here finite element analysis approach to obtain 3-D temperature distribution in heating with a four-applicator array. A comparison of phantom and simulation temperatures indicated that inclusion of shear component in heating with ultrasound interstitial applicators improves significantly the simulation quality factor. We included this component in simulations of 3-D temperature pattern when heating brain tissue with the array. The simulations for the tissue show that the perfusion remains a primary factor in defining the pattern.

Ultrasonic waveguide applicator arrays for interstitial heating: a model study.

In this paper we describe an ultrasonic waveguide multiapplicator array for interstitial heating. We first discuss the heat generation term common for this type of applicator and show that the radius of the applicator is the limiting factor in the pattern of heat deposition. We carry out finite element analysis simulations of temperature profiles for three- and four-applicator array, and we test the simulations by measurements in a large volume tissue phantom. With the positive result of this test, we use the simulations to evaluate the size of the heated volume for several applicators (three to six) and for various geometries of their positioning. We do the simulations for a range of the effective thermal conductivity and for two applicator diameters. The volume of the medium with temperatures above 42 degrees C was in the 25 to 73 cm(5 ) range. This volume increased linearly with the diameter of the boundary at the basal temperature. Power required to produce preselected temperature elevation increased monotonically with the effective thermal conductivity. With the 24 mm between the applicators, the array could elevate the temperature to the required value up to the 0.030 W/cm/K effective thermal conductivity.

[Epidemiologic and clinical tests of patients suffering from laryngeal cancer in the Lublin macro-region in the years 1991-1995]. / Badania epidemiologiczne i kliniczne chorujacych na raka krtani w latach 1991-1995 w makroregionie lubelskim.

The study of registered morbidity of patients suffering from laryngeal cancer in the Lublin macro-region in the years 1991-95 was carried out. Morbidity rates in the population in consecutive years of investigated 5-year age groups were examined. In clinical investigations the conditions of clinical advancement of the disease in particular organs was determined and environmental factors and occupation were also revealed.

Feasibility of ultrasound hyperthermia with waveguide interstitial applicator.

Interstitial and intracavitary ultrasonic hyperthermia applicators facilitate well-controlled power deposition in tissues. In this paper, analysis of temperature elevation and experimental results in tissue phantom, animal tissue in vivo and animal tissue in vitro are presented for a waveguide applicator intended for treatment of brain tumors. It consisted of a G18 hypodermic needle attached via a conical velocity transformer to a 12.7-mm-diameter piezoelectric disk operated at 1.0 MHz. The axial acoustic pressure distribution had a standing-wave pattern with the four cycles/cm spatial periodicity. This periodicity was absent in the temperature distribution in tissue phantoms. The simulations based on a solution to the effective heat conductivity equation indicated that the hyperthermic range can be reached within a 4- and a 10-mm radius around the applicator for a 21- and a 60-mm sample diameters, respectively, with reasonable input power. The first number corresponded closely to the 5-mm radius observed in porcine brain in vivo and the second one came close to the 9-mm radius in porcine brain in vitro. The presented results demonstrate the potential of the ultrasound waveguide interstitial applicator for hyperthermia of small volume tumors.

Variability of modes of division of the superior lobar bronchi in the right and left lungs.

The studies were carried out on 100 right lungs and 100 left lungs taken from cadavers of humans of both sexes, aged 18 to 80 years. After swilling, the bronchi were injected with 65% duracryl and after 24 hours digested in sulphuric acid. The obtained casts were measured and the modes of division of superior lobar bronchi were determined. The results of observation and measurements were presented in the form of schematic drawings and documented with photographs. On the basis of the modes of branching of superior lobar bronchi three types and eight variants of division for the right lobar bronchus and two types and five variants for the left lobar bronchus were identified. The divergence between the frequency of some modes of division of the studied bronchi, especially according to authors from other countries and very similar results of investigations relating to atypical modes of division of some bronchi, performed in Poland and neighbouring countries, were noted.

[Weight (mass) of the thyroid in assessment of endemic goiter]. / Ciezar (masa) tarczycy w ocenie endemii wola.

The weight of thyroids from cadavers in the Province of Wroclaw in the year 1991 was not lower and, on the contrary it was frequently higher than in the year 1957. One of the causes of this may be incorrectly conducted iodine prophylaxis. Other causes cannot be ruled out including the influence of pollution after Chernobyl reactor disaster. The data from the Province of Lublin indicate that mass of thyroids in this region is even higher than that in the Province of Wroclaw considered to be the area of goitre endemic.