Progenitor cells are responsible for formation of human prostate epithelium primary cultures

To analyze cell viability and morphology of primary cell cultures from CD133 immunolabeled and sorted cells from epithelium of patients suffering from benign prostate hyperplasia (BPH). Methods: Cells obtained from 5 patients were divided in two fractions. First fraction (CD133+/CD133–) was cultivated in DMEM with 10% FBS. Second fraction was mixed with CD133 microbeads and immunomagnetically divided into CD133+ and CD133– fractions. These cells were cultivated and followed-up for 2 weeks. Cells were stained for Annexin V FITC/propidium iodide. Results: Seventy CD133+/CD133– cultures, thirty-one of CD133+ and thirty-one of CD133– cells were established. There were 5-fold and 3-fold increase of CD133+/CD133- and CD133+ cell number after 2 weeks, respectively. CD133+/CD133– and CD133+ monolayers displayed epithelial-like morphology and cytokeratine expression. CD133– cultures collapsed. Cell viability within CD133+ and CD133– populations was 90.1 ± 6.3% and 24.3 ± 6.2%, respectively. Apoptotic index was 9.0 ± 6.1% and 28.5 ± 23.8% within CD133+ and CD133– cultures, respectively. Conclusions: CD133 separated human primary epithelial cell cultures displayed differences in morphology, viability and apoptosis occurrence. Immunomagnetic sorting can be recommended in each in vitro experiments with primary cell cultures in order to provide more objective results.Цель: оценить жизнеспособность и морфологию клеток первичных клеточных культур, полученных из меченных по CD133 и полученных с помощью клеточной сортировки клеток эпителия пациентов с доброкачественной гиперплазией предстательной железы (BPH). Методы: клетки, полученные от 5 пациентов, были разделены на 2 фракции. Первую фракцию (CD133+/CD133–) выращивали в DMEM с 10% FBS али с CD133 магнитными гранулами и с помощью магнита разделили клетки на CD133+- и CD133–-фракции. Далее клетки культивировали в течении 2 нед. Клетки окрашивали аннексиномV FITC/пропидий йодидом. Результаты: получено 70 CD133+/CD133- -культур клеток, 31 CD133+ и 31 CD133–. Через 2 нед культивирования отмечали 5-кратное и 3-кратное увеличение количества CD133+/CD133– и CD133+ клеток соответственно. CD133+/CD133- -и CD133+-клетки росли в монослое и имели морфологию эпителиальных клеток, экспрессировали цитокератин. CD133–-клетки не выжили. Выживаемость клеток в популяциях CD133+ и CD133– была 90,1 ± 6,3% и 24,3 ± 6,2% соответственно. Показатель апоптического индекса для культур CD133+ и CD133– был 9,0 ± 6,1% и 28,5 ± 23,8% соответственно. Выводы: показаны различия в морфологии, выживаемости клеток и частоте апоптоза для эпителиальных клеток, разделенных в зависимости от экспрессии CD133. Сортировка клеток с помощью иммуномагнитного разделения рекомендована для каждого in vitro эксперимента с использованием первичных клеточных культур для получения более объективных результатов

Urological procedures in Central Europe and the current reality based on the national registries of Czech Republic, Hungary, and Poland (2012 status)

INTRODUCTION: In recent years, the laparoscopic approach in oncologic urology seems more attractable to the surgeons. It is considered to have the same oncologic quality as open surgery, but is less invasive in patients. It is used widely in all of Europe, but with various frequency. The aim of the study was to present a various amount of oncourological procedures from three neighbouring countries - Poland, Czech Republic and Hungary. Prostatectomy, cystectomy, nephrectomy and tumorectomy (Nephron Sparing Procedures - NSS) were presented as a list of procedures prepared from the national registry. MATERIAL AND METHODS: The total amount of procedures was presented, as well as the LO (Lap to Open procedures) index, P/P (procedures/population) index, ratio of cystectomy/population, and cystectomy/TURBT. RESULTS: In the Czech Republic, the most complex procedures are performed (laparoscopic/robotic prostatectomy, NSS LAP, LAP nephrectomy) in the majority when analysing the country's population. In Hungary and Czech Republic, there are more laparoscopic/robotic radical prostatectomies performed, than open ones. In Poland the largest number of cystectomies is performed when analysing the country's population, but it is difficult to explain the much higher ratio of 6.57 TUR/one cystectomy. In the Czech Republic this procedure is performed in almost one quarter of the patients (23.36%). Interestingly, in Hungary the cystectomy with pouch creation is performed in about 67.65% cases. The highest reimbursement for surgical procedure is present in the Czech Republic with approximately 20-40% more than when compared to Poland or Hungary. CONCLUSIONS: The definitive leader in Central Europe (based on the national registry) is the Czech Republic, where the most complex procedures are performed (laparoscopic/robotic prostatectomy, NSS LAP, LAP nephrectomy) in biggest amounts when analysing the country's population. Explanation of such circumstances, can be the higher reimbursement rate for surgical procedure in this country

Influence of crude oil and detergents on the concentration of some ions in hemolymph of the shrimp Cragon cragon L. and the crab Rhithropanopeus harrisi Gould

Solutions of detergent or crude oil of 10 ppm, 50 ppm, and 100 ppm after 48-96 hours of exposure induce significant changes in the ionic composition of the hemolymph of the shrimp Crangon crangon and the crab Rhithropanopeus harrisi. The concentrations of sodium, calcium, and chloride ions decrease, whereas the concentration of potassium increases in both species exposed to both pollutants. - The magnesium ion concentration decreases in the hemolymph of the shrimp incubated in solutions of detergent and in the crab incubated in the water containing crude oil. - lt is suggested that changes of the ionic composition of the hemolymph of the studied shrimps and crabs under influence of both pollutants are possibly brought about by alteration of cell membrane permeability and action on some enzyme activities

Prostate cancer which affects an elderly man is a feature of senescence (cellular) — a biology phenomenon

The aim of this paper was to present late-life form of the prostate cancer, which differs from its aggressive counterpart that affects men between 55–65 years old and younger. The differences can be found in carcinogenesis risk factors, cancer biology and finally patients’ survival. The most important is that these two clinical (age-related) forms of the prostate cancer are still undistinguishable in clinico-pathology reports and patients bearing different diseases are offered the same treatment. Potential mechanisms leading to development of the late-life clinically indolent prostate cancer are discussed. It seems that the key abnormalities are proteins involved in control of regenerative potential and cell senescence

Purification and properties of soluble arylsulphatases isolated from hepatopancreas of the shrimp Crangon crangon L.

Arylsulphatase A and B were isolated from the hepatopancreas of the shrimp Crangon crangon L. after 7 days incubation in 50 ppm of detergent "SOLO". Arylsulphatase A, B-1 and B-2 were isolated from the hepatopancreas of the shrimp C. crangon L. incubated in pure brackish water (salinity 7‰). - Heavy and light fuel oil added to the enzyme in vitro in conc. of 2.0% inhibit the activity of arylsulphatase A in greater degree than arylsulphatase B (14.4% and 5.5% respectively). - Detergent "SOLO" (mixture of nonionic and anionic detergents) in the conc. of 0.5% inhibits for 58% arylsulphatase A and for 91% arylsulphatase B, whereas in conc. of 2.0% it inhibits arylsulphatase A for 91.7% and arylsulphatase B for 100%

Influence of two pt(iv) complexes on viability, apoptosis and cell cycle of B16 mouse melanoma tumors

Several platinum(IV) complexes are showing considerable promise in initial trials, producing reactive intermediates that then interact with DNA. Aim: To perform in vitro study of two new platinum(IV) complexes cytotoxic effect on B16 mouse melanoma cells. Methods: PtCl₄ (dbtp)₂ and PtCl₂ (6mp)₂ complexes were prepared. PtCl₄ (dbtp)₂ was created as modification of PtCl₄ (dmtp) test previously.Apoptosis and necrosis were examined using flow cytometry, upon Annexin V/PI staining. Results: LC₁₀,LC₅₀ andLC₉₀ parameters established for PtCl₄ (dbtp)₂ were as following: 2.6, 17.0, 58.0 μmol/L. However LC₁₀ andLC₅₀ established for PtCl₂ (6mp)₂ were 1.2 and 14.0μmol/l respectively. The both complexes induced apoptosis. PtCl₂ (6mp)₂ induced cell cycle arrest in G0/G1, while PtCl₄ (dbtp)₂ — in S-phase. Conclusions: PtCl₄ (dbtp)₂ appeared to be more cytotoxic against B16 cells than PtCl₂ (6mp)₂ . Apoptosis was the main mechanism of cell loss in cultures incubated with both tested complexes

Cancer Biomarker Discovery: The Entropic Hallmark

Background: It is a commonly accepted belief that cancer cells modify their transcriptional state during the progression of the disease. We propose that the progression of cancer cells towards malignant phenotypes can be efficiently tracked using high-throughput technologies that follow the gradual changes observed in the gene expression profiles by employing Shannon's mathematical theory of communication. Methods based on Information Theory can then quantify the divergence of cancer cells' transcriptional profiles from those of normally appearing cells of the originating tissues. The relevance of the proposed methods can be evaluated using microarray datasets available in the public domain but the method is in principle applicable to other high-throughput methods. Methodology/Principal Findings: Using melanoma and prostate cancer datasets we illustrate how it is possible to employ Shannon Entropy and the Jensen-Shannon divergence to trace the transcriptional changes progression of the disease. We establish how the variations of these two measures correlate with established biomarkers of cancer progression. The Information Theory measures allow us to identify novel biomarkers for both progressive and relatively more sudden transcriptional changes leading to malignant phenotypes. At the same time, the methodology was able to validate a large number of genes and processes that seem to be implicated in the progression of melanoma and prostate cancer. Conclusions/Significance: We thus present a quantitative guiding rule, a new unifying hallmark of cancer: the cancer cell's transcriptome changes lead to measurable observed transitions of Normalized Shannon Entropy values (as measured by high-throughput technologies). At the same time, tumor cells increment their divergence from the normal tissue profile increasing their disorder via creation of states that we might not directly measure. This unifying hallmark allows, via the the Jensen-Shannon divergence, to identify the arrow of time of the processes from the gene expression profiles, and helps to map the phenotypical and molecular hallmarks of specific cancer subtypes. The deep mathematical basis of the approach allows us to suggest that this principle is, hopefully, of general applicability for other diseases