Dietary carotenoids in normal and pathological tissues of corpus uteri.

Carotenoids and retinyl esters are the source of vitamin A in the human body and its natural derivatives takes part in the regulation of cell replication and differentiation in the human endometrium, may induce the leiomyoma growth and has a role in differentiation of endometrial adenocarcinoma. The aim of the study was to demonstrate the presence of carotenoids in tissues from the normal uterus and from various tumors of the uterine corpus, as well as to compare the total content, major carotenoids and % of carotenoids belonging to the provitamin A group between the tissues examined. Using three independent methods of chromatography (CC, TLC, HPLC) we analysed 140 human samples. We identified 13 carotenoids belonging to the eg. provitamin A group and epoxy carotenoids. In all the samples beta-carotene, beta-cryptoxanthin, lutein, neoxanthin, violaxanthin and mutatoxanthin were isolated. In normal tissues, the mean carotenoid content was the highest in the follicular phase endometrium (9.9 microg/g), while the highest percentage of carotenoids belonging to provitamin A group was found in the luteal phase (18.2%). In the pathological group, the highest mean values were demonstrated for epithelial lesions (8.0 microg/g), and within this group - in endometrioid adenocarcinoma (10.8 microg/g). In both groups, violaxanthin, beta-cryptoxanthin, lutein epoxide and mutatoxanthin were the predominant carotenoids. We have demonstrated that all uterine tissues show a concentration of beta-carotene and beta-cryptoxanthin, being the source of vitamin A. The highest total values of carotenoids obtained in the group of endometrioid adenocarcinoma seem to confirm certain enzymatic defects in carotenoid metabolism in the course of the neoplastic process or some metabolic modifications. The finding of astaxanthin - the major antioxidant among carotenoids - in 63% of tissues examined is also significant

Is the repair of articular cartilage lesion by costal chondrocyte transplantation donor age-dependent? An experimental study in rabbits.

The repair of chondral injuries is a very important problem and a subject of many experimental and clinical studies. Different techniques to induce articular cartilage repair are under investigation. In the present study, we have investigated whether the repair of articular cartilage folowing costal chondrocyte transplantation is donor age-dependent. Transplantation of costal chondrocytes from 4- and 24-week old donors, with artificially induced femoral cartilage lesion, was performed on fourteen 20-week-old New Zealand White male rabbits. In the control group, the lesion was left without chondrocyte transplantation. The evaluation of the cartilage repair was performed after 12 weeks of transplantation. We analyzed the macroscopic and histological appearance of the newly formed tissue. Immunohistochemistry was also performed using monoclonal antibodies against rabbit collagen type II. The newly formed tissue had a hyaline-like appearance in most of the lesions after chondrocyte transplantation. Positive immunohistochemical reaction for collagen II was also observed in both groups with transplanted chondrocytes. Cartilage from adult donors required longer isolation time and induced slightly poorer repair. However, hyaline-like cartilage was observed in most specimens from this group, in contrast to the control group, where fibrous connective tissue filled the lesions. Rabbit costal chondrocytes seem to be a potentially useful material for inducing articular cartilage repair and, even more important, they can also be derived from adult, sexually mature animals

Combined therapy with disintegrin and melphalan as a new strategy in inhibition of endometrial cancer cell line (Ishikawa) growth.

Endometrial cancer is one of the most frequently diagnosed cancer in females with prevalence of 22 in 100,000 women. The etiology of the cancer remains unclear. Despite significant progress towards understanding the patho-mechanism of the disease, effective treatment is still lacking. The results of the study suggest that combined treatment of Ishikawa cells for 24 h with disintegrin and then for 24 h with melphalan severely inhibits important biological functions of the cells. We showed that such strategy have a potent cytotoxic effect. The mechanism of process undergoes probably through inhibition of integrin - dependent signaling. In this study we shown down regulation of Shc and FAK proteins in cells treated with echistatin and melphalan. It suggests that signaling pathways that involve Shc and FAK participation may represent target for antineoplastic strategy. The functional significance of the combined treatment of Ishikwa cells with echistatin and melphalan was found at the level of collagen biosynthesis. Decreased biosynthesis of collagen in extracellular matrix may suppress cell growth and induce apoptosis. The treatment with echistatin and melphalan also showed decreased expression of IGF receptor in comparison to the cells treated with both compounds separately. The data presented suggest that combined therapy with disintegrin - echistatin and alkyalting drug - mephalan may represent a new approach to more effective and safe cancer therapy

Dietary carotenoids in normal and pathological tissues of corpus uteri.

Carotenoids and retinyl esters are the source of vitamin A in the human body and its natural derivatives takes part in the regulation of cell replication and differentiation in the human endometrium, may induce the leiomyoma growth and has a role in differentiation of endometrial adenocarcinoma. The aim of the study was to demonstrate the presence of carotenoids in tissues from the normal uterus and from various tumors of the uterine corpus, as well as to compare the total content, major carotenoids and % of carotenoids belonging to the provitamin A group between the tissues examined. Using three independent methods of chromatography (CC, TLC, HPLC) we analysed 140 human samples. We identified 13 carotenoids belonging to the eg. provitamin A group and epoxy carotenoids. In all the samples beta-carotene, beta-cryptoxanthin, lutein, neoxanthin, violaxanthin and mutatoxanthin were isolated. In normal tissues, the mean carotenoid content was the highest in the follicular phase endometrium (9.9 microg/g), while the highest percentage of carotenoids belonging to provitamin A group was found in the luteal phase (18.2%). In the pathological group, the highest mean values were demonstrated for epithelial lesions (8.0 microg/g), and within this group - in endometrioid adenocarcinoma (10.8 microg/g). In both groups, violaxanthin, beta-cryptoxanthin, lutein epoxide and mutatoxanthin were the predominant carotenoids. We have demonstrated that all uterine tissues show a concentration of beta-carotene and beta-cryptoxanthin, being the source of vitamin A. The highest total values of carotenoids obtained in the group of endometrioid adenocarcinoma seem to confirm certain enzymatic defects in carotenoid metabolism in the course of the neoplastic process or some metabolic modifications. The finding of astaxanthin - the major antioxidant among carotenoids - in 63% of tissues examined is also significant

Zdrowie dzieci z ciąż po leczeniu technikami rozrodu wspomaganego medycznie

Since the birth of Louise Brown in July 1978 and the birth of the first intracytoplasmic sperm injection (ICSI) child in January 1992 many couples with female-factor or male-factor infertility can be helped to overcome their infertility resulting in a delivery and birth of a child. Over a million children have been born from assisted conception worldwide. Newer techniques being introduced appear less and less ‘natural’, such as intracytoplasmic sperm injection (ICSI), but there is little information on these children beyond the neonatal period. This risk varied according to the patient’s age, the type of ART procedure performed, the number of embryos transferred, and embryo availability. ART is associated with low increase risk of congenital malformations, major birth defects and genetic and imprinting disorders.Szacuje się, że od urodzenia w 1978 roku Louise Brown i dziecka po ICSI bardzo dużo niepłodnych par zarówno z czynnikiem męskim jak i żeńskim może pokonać niepłodność i urodzić własne dziecko. Na całym świecie urodziło się ponad milion dzieci dzięki zastosowaniu technik rozrodu wspomaganego medycznie. Najnowsze techniki stają się coraz mniej naturalne, tak jak ICSI, ale wciąż mało jest informacji o dzieciach po ART. Wydaje się, że ryzyko urodzenia dziecka z problemami zdrowotnymi zależy od wieku przyszłych rodziców, rodzaju stosowanej procedury ART, liczby i jakości zarodków. ART są związane z nieznacznie zwiększonym ryzykiem wystąpienia wad wrodzonych i dużych defektów zaburzeń genetycznych i epigenetycznych

Brak korelacji stężeń amoniaku i żeńskich hormonów płciowych w ludzkim przedowulacyjnym płynie pęcherzykowym

Objective: The ammonia gradient from the human preovulatory follicular fluid (FF) to blood has been documented. The purpose of the present study was to substantiate whether, following controlled ovarian hyperstimulation, female sex hormones are related to this phenomenon. Material and Methods: Blood was taken from the antecubital veins of 32 randomly selected patients undergoing an in vitro fertilization program prior to oocyte retrieval and FF collection. Ammonia concentrations in blood and FF were determined by the indophenol method, and 17β-estradiol (E2) and progesterone (PGS) concentrations In plasma and FF by radioimmunoassay. Results: The mean ammonia concentration was 39.15 +/-3.25 μM for FF, and 20.15 +/-1.20 μM for blond (pCel: Udokumentowano istnienie gradientu stężeń amoniaku między ludzkim przedowulacyjnym płynem pęcherzykowym (FF) a krwią. Celem niniejszej pracy było ustalenie czy – po kontrolowanej hiperstymulacji jajników – żeńskie hormony płciowe mają związek z tym zjawiskiem. Materiał i metoda: Krew żylną ze zgięcia łokciowego pobrano bezpośrednio przed pozyskaniem oocytu i FF od 32 losowo wybranych pacjentek poddanych procedurze zapłodnienia pozaustrojowego. Stężenia amoniaku we krwi pełnej i w FF zmierzono metodą indofenolową, a stężenia 17β-estradiolu (E2) i progesteronu (PGS) w osoczu i w FF – metodą radioimmunologiczną. Wyniki: Średnie stężenie amoniaku wyniosło 39,15 +/- 3,25 μM w FF, a 20,15 +/-1,20 μM we krwi (p < 0,001). U wszystkich badanych stosunek stężenia amoniaku w FF do stężenia amoniaku we krwi wyniósł powyżej 1,0, co potwierdza produkcję amoniaku przez pęcherzyk przedowulacyjny. Nie stwierdzono istnienia korelacji między stężeniami amoniaku i E2 w FF (współczynnik korelacji rang Spearmana r = 0,2546; p = 0,160), ani między stężeniem amoniaku w FF i różnicą stężeń E2 między FF a osoczem (r = 0,2416; p = 0,183). Podobnie nie wykryto korelacji między stężeniami amoniaku i PGS w FF (r = -0,1089; p = 0,553), co podkreślił brak korelacji między stężeniem amoniaku w FF i różnicą stężeń PGS między FF a osoczem. Wnioski: Produkcja amoniaku nie jest bezpośrednio powiązana z wewnątrzpęcherzykowymi stężeniami żeńskich hormonów płciowych. Natomiast wewnątrzpęcherzykowa akumulacja amoniaku może być odpowiedzialna za zasadowe pH ludzkiego płynu pęcherzykowego

Rabbit articular cartilage defects treated with cultured costal chondrocytes (preliminary report)

An attempt to repair articular cartilage defects by costal chondrocytes transplantation was made. A full-thickness defect in the rabbit's femoral patellar groove was artificially made. Cultured costal cartilage chondrocytes were then transplanted into the defects and covered with periosteal flaps. Empty defects were used as the control group. Animals were divided into two groups (five rabbits each). They were examined after four and twelve weeks from the day of transplantation, respectively. The reparative tissue was evaluated by macroscopic and histological examinations. The reparative tissues in defects with transplanted chondrocytes had an hyaline-like cartilage appearance and were firmly attached to the surrounding normal cartilage. No trace of newly formed bone was detected. The reparative tissues found in defects that were left empty had a fibrous character. They were loosely connected to the surrounding cartilage and were more compliant than tissues from transplanted defects. Considering these initial findings, the ease of surgical procedures during the harvesting of the costal cartilage and few interventions into the joint make the costal cartilage a promising source of chondrocytes for transplantation. However, this needs to be confirmed on a larger scale over a longer period of time

The effect of doxorubicin and retinoids on proliferation, necrosis and apoptosis in MCF-7 breast cancer cells.

Doxorubicin (Adriamycin) is the most active drug in the treatment of breast cancer. The aim of this study was to investigate the interaction of doxorubicin and retinoids in the inhibition of proliferation of hormone sensitive (ER+) human breast cancer cell line MCF-7 and to find out whether this combination can result in the enhancement of its therapeutic effect. As a comparison we also used estradiol and tamoxifen. We also made an attempt to elucidate the effect of these compounds on the stimulation of the apoptotic pathway in breast cancer cells. Cell proliferation in a 24-hour culture was assessed by [3H] thymidine incorporation into cancer cells and by immunocytochemical analysis of cellular cycle-related PCNA and Ki-67 antigens expression, after the incubation of the cell culture with 10, 20 and 50 nM doxorubicin (DOX), 2 nM estradiol (E2), 10 microM tamoxifen (TAM) and 1 nM, 0.01, 0.1, 1 and 10 microM of all-trans retinoid acid (ATRA). The assessment of cell viability and analysis of apoptotic and necrotic cells were performed after the 72-hour incubation of the culture with the examined substances and following apoptosis induction using acridine orange and ethidine bromide. Of the doxorubicin concentrations used in the study, 20 nM inhibited thymidine incorporation to 84.83 +/- 10.00% (control=100%). In the same culture conditions, 2 nM E2 stimulated cancer cells to 157.09 +/- 8.84%. Concentrations of 10 microM TAM and 10 microM ATRA inhibited the proliferation to 63.16 +/- 7.85% and 52.19 +/- 3.21%, respectively. A statistically significant reduction of these values was observed when 20 nM DOX was added to medium with E2 - 39.24 +/- 7.6%, TAM - 48.34 +/- 2.05% and ATRA - 21.98 +/- 1.69%, respectively; the percentage of PCNA- and Ki-67-positive cells was also reduced. Despite high antiproliferative efficacy of 20 nM DOX and 10 microM ATRA combination, the percentage of apoptotic cells was only 25 +/- 0.81%, being similar to that obtained in the culture with 20 nM DOX. The concentrations of 10, 20 and 50 nM DOX that were used to inhibit the proliferation of MCF-7 cell line were not particulary effective. The inhibitory effect was obtained when 20 nM of DOX and E2, TAM or ATRA were used simultaneously. The use of E2 caused a two-fold decrease in the percentage of proliferating cells. It was also shown that the effectiveness of DOX in combination with ATRA is significantly higher than that of DOX combined with TAM, which might suggest a valuable approach to the treatment of breast cancer

Altered Peroxisome-Proliferator Activated Receptors Expression in Human Endometrial Cancer

Peroxisome proliferator-activated receptors (PPARs) belong to a family of nuclear hormone receptors acting as transcriptional factors, recently involved also in carcinogenesis. Present study was undertaken to evaluate the presence and subcellular localization of different PPAR isoforms (α, β, γ) in healthy endometrial tissue (n = 10) and endometrial carcinoma (FIGO I, endometrioides type, G1, n = 35). We sought to analyze PPARs mRNA content as well as protein immunohistochemical expression that was further quantified by Western Blot technique. For both PPARα and PPARβ, protein expression was significantly higher in endometrial cancers compared to normal endometrial mucosa. In opposite, PPARγ protein expression was lower in endometrial cancer cells. In each case, immunohistochemical reaction was confined to the perinuclear and/or nuclear region. At the transcriptional level, the content of mRNA of all PPAR subunits did not follow the protein pattern of changes. These results provide evidence for altered PPAR's protein expression and disregulation of posttranslational processes in endometrial cancers