Lysophosphatidic acid and sphingosine 1-phosphate metabolic pathways and their receptors are differentially regulated during decidualization of human endometrial stromal cells.

In the luteal phase, human endometrial stromal cells (HESCs) undergo proliferation, migration and differentiation during the decidualization process under the control of the ovarian steroids progesterone and estrogen. Proper decidualization of stromal cells is required for blastocyst implantation and the development of pregnancy. The proliferation, migration and differentiation of HESCs in decidualization do not require the presence of a blastocyst but are greatly accelerated during implantation. Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are potent bioactive lysophospholipids that have critical roles in various physiological and pathophysiological processes, including inflammation, angiogenesis and cancer. The expression of the enzymes involved in LPA and S1P turnover and their receptors in HESCs during decidualization has not been characterized yet. We found that the LPAR1 and LPAR6 and S1PR3 receptors are highly expressed in HESCs. LPAR1, autotaxin (ATX), an LPA producing enzyme and lipid phosphate phosphatase 3 were up-regulated during decidualization. Interestingly, the expression of all S1P receptor subtypes and LPA receptors (LPAR2-6) mRNA was down-regulated after decidualization. We found that SPHK1 is highly expressed in HESCs, and is up-regulated during decidualization. S1P phosphatase SGPP1 and S1P lyase SGPL1 are highly expressed in HESCs. SGPP1 mRNA expression was significantly up-regulated in decidualized HESCs. In conclusion, this study shows the first time that specific LPA and S1P receptors and their metabolizing enzymes are highly regulated in HESCs during decidualization. Furthermore, we suggest that LPAR1 receptor-mediated signaling in HESCs may be crucial in decidualization process. SPHK1 activity and high turnover of S1P and LPA might be essential for precise regulation of their signaling during decidualization of human endometrium and implantation.

The effect of prolactin and estrogen cross-talk on prolidase- dependent signaling in MCF-7 cells.

Estrogen and prolactin play important role in mammary carcinogenesis. The present study was undertaken to evaluate the effect of prolactin and estrogen cross-talk on HIF-1 α level and expression of some HIF-1 α- dependent signaling proteins. Since up-regulation of prolidase activity inhibits HIF-1 α degradation, the enzyme was considered as an interface of estrogen/prolactin signaling. The experiments were performed on MCF-7 cells cultured with prolactin in the presence or absence of estradiol. It was found that in the presence of estradiol, prolactin inhibits prolidase activity and its down-stream signaling proteins: HIF-1α, mTOR, AKT and MAPK p-38, while in the absence of estradiol, an opposite effect was observed. These results suggest that prolactin/estrogen cross-talk exert beneficial effect on prolidase-dependent down regulation of HIF-1α. It suggests that dual action of prolactin and estrogen may be considered as a strategy in therapy of breast cancer.

Evidence that FGFR1 loss-of-function mutations may cause variable skeletal malformations in patients with Kallmann syndrome.

PURPOSE: Loss-of-function mutations in FGFR1 have been identified in approximately 10% of the Kallmann syndrome (KS) patients. Previous reports have focused mainly on olfactory, reproductive, and some other features such as cleft lip/palate and dental agenesis. Given the ubiquitous expression of FGFR1 during development, other abnormal phenotypes might, however, have been overlooked in these patients. Here, we demonstrate skeletal phenotypic characterization of patients presented with KS and FGFR1 mutations. MATERIAL AND METHODS: Using the Sanger DNA sequencing technique a cohort of 29 KS patients was screened. RESULTS: Here, we report on 5 KS patients who carry FGFR1 mutations (Gly270Asp, Gly97Ser, Met161Thr, Ser685Phe and Ala167Ser/Ala167Ser). Three patients presented with skeletal abnormalities, i.e. spine (hemivertebra and butterfly vertebra) and limb (oligodactyly of the feet, fusion of the 4th and 5th metacarpal bones) malformations in two patients and one patient, respectively. The hand phenotype found in the patient cannot be thought of as a counter-type of the hand phenotype resulting from FGFR1 gain-of-function mutations. The skeletal anomalies identified in the 3 KS patients are close to those observed in Fgfr1 conditional knockout mice. CONCLUSIONS: This study demonstrates that FGFR1 loss-of-function mutations can be associated with skeletal abnormalities also in humans. Further investigations in KS patients who carry FGFR1 mutations are needed to evaluate the prevalence of skeletal defects in this genetic form of KS. Conversely, the presence of bone malformations in a KS patient should direct the geneticist towards a search for mutations in FGFR1.

Increased serum level of membrane type 1-matrix metalloproteinase (MT1-MMP/MMP-14) in patients with breast cancer.

UNLABELLED: Different types of matrix metalloproteinases, including membrane type 1 matrix metalloproteinase (MT1-MMP/MMP-14) can be easily detected in biological fluids and therefore may be contemplated as putative tumor markers. Although increased activity of MT1-MMP/MMP-14 have already been found in breast cancer, little is known about its circulating levels. The aim of the present study was therefore to evaluate serum levels of active form of membrane type 1 matrix metalloproteinase (MT1-MMP/MMP-14). A novel type of activity enzyme-linked immunosorbent assay was used to detect serum levels of MT1-MMP/MMP-14 in 18 patients with invasive ductal breast cancer and 11 healthy controls. In the breast cancer group of patients MT1-MMP/MMP-14 mean (+/-SD) concentration was 16.91+/-5.87 ng/ml which was significantly higher (p<0.0001) than the mean values obtained for the control i.e. 8.55+/-1.66 ng/ml. CONCLUSIONS: Higher levels of soluble form of MT1-MMP/MMP-14 could play a role in invasiveness and metastasis of breast cancer. Whether or not it has a potential as biochemical marker remains to be determined.

Chlamydia trachomatis urogenital infection in women with infertility.

PURPOSE: The study objective was to evaluate the prevalence of urogenital Chlamydia trachomatis (C.tr.) infection in women with diagnosed infertility. MATERIAL AND METHODS: The study involved patients from the Department of Gynecological Endocrinology and from the Center for Reproductive Medicine "Kriobank" in Bialystok. Female patients (n=71), aged 23-41, were divided into two groups according to the main diagnosis: A--tubal infertility (23) and B--infertility of another origin (48). For direct testing, PCR method was used to detect C.tr. infection in cervical samples (Roche, Molecular Systems, N.J., USA). Specific IgA and IgG anti-chlamydial antibodies in the serum were determined by immunoenzymatic assay (medac, Hamburg, Germany). Diagnostic procedures were performed at the Centre for STD Research and Diagnostics in Bialystok. RESULTS: In group A, C.tr. infection was detected in: 8.7% patients, in group B--8.3%. Specific anti-C.tr. antibodies IgA were detected in: 13.0% in group A and 6.3% in group B, IgG respectively in 39.1% and in 10.4%. CONCLUSIONS: 1. C.tr. infection is very important etiological factor of female infertility. 2. The detection of specific antichlamydial antibodies is a valuable, noninvasive diagnostic procedure. 3. Infertile women should be routinely tested for C.tr. infection.

The FTO gene modifies weight, fat mass and insulin sensitivity in women with polycystic ovary syndrome, where its role may be larger than in other phenotypes.

AIM: Genome-wide association studies have shown that variation in the FTO gene predisposes to obesity and related traits that are common features of polycystic ovary syndrome (PCOS). The aim of the present study was to assess the effect of FTO variation on obesity, insulin sensitivity, and metabolic and hormonal profiles in PCOS. METHODS: We examined 136 PCOS women (mean body mass index [BMI]: 28.28+/-6.95kg/m(2), mean age: 25.36+/-5.48 years). Anthropometric measurement, euglycaemic-hyperinsulinaemic clamp and oral glucose tolerance tests and sex hormone assessments were performed. The study group was genotyped for the FTO rs9939609 polymorphism. RESULTS: BMI (29.0+/-6.9kg/m(2) vs 26.1+/-6.8kg/m(2); P=0.023), body weight (80.1+/-20.7kg vs 72.6+/-20.2kg; P=0.048), fat mass (29.7+/-1 6.6kg vs 24.6+/-17.7kg; P=0.045) and waist circumference (89.8+/-16.7cm vs 83.2+/-17.1cm; P=0.028) were higher in carriers of at least one copy of the A allele. Differences in these parameters were more significant when comparing AA and TT homozygotes. Women with the AA genotype also had decreased insulin sensitivity (P=0.025) and follicle-stimulating hormone (P=0.036). In logistic-regression analyses, the association of the FTO gene polymorphism with insulin sensitivity was no longer significant when BMI was included in the model. CONCLUSION: Variation in the FTO gene modifies weight, adiposity and other measures of obesity and insulin sensitivity in PCOS. The examined FTO gene variant appears to have a greater impact on obesity and related traits in PCOS than in other phenotypes. The effect on insulin sensitivity appears to be secondary to its influence on obesity and body fat.

Estrogen-dependent regulation of PPAR-gamma signaling on collagen biosynthesis in adenocarcinoma endometrial cells.

The link between estrogen and metabolic developmental factors of endometrial carcinoma is well established. PPAR- gamma, (an important modulator of metabolism) and estrogen receptor belong to a family of nuclear hormone receptors that were shown to interact with each other. The interaction may affect transcriptional activity of these transcription factors. The anti-diabetic troglitazone (TGZ) is well known PPAR- gamma ligand. The effect of troglitazone-induced PPAR- gamma activation on estrogen-dependent stimulation of collagen biosynthesis was studied in the Ishikawa endometrial adenocarcinoma cell line. We have found that the presence of estrogen activity in growth medium (1nM) augmented collagen biosynthesis in the cells. An addition of PPAR- gamma agonists, as troglitazone or clofibrat to the growth medium induced inhibition of collagen biosynthesis. The inhibition was effective only when estrogen receptor was stimulated, since removal of estrogen receptor by ICI 182- 780-dependent degradation did not affect collagen biosynthesis. The mechanism of the inhibition was found at the level of NF-kB (known inhibitor of collagen gene expression) and MAPK signaling. PPAR- gamma ligands stimulated expression of NF-kB, while they inhibited expression of p-38 but not ERK1/ERK2. The data document for the first time that inhibitory effect of PPAR- gamma ligands on collagen biosynthesis in endometrial adenocarcinoma cells requires functional estrogen receptor.

Estradiol and tamoxifen differently affects the inhibitory effects of vitamin A and their metabolites on the proliferation and expression of alpha2beta1 integrins in MCF-7 breast cancer cells.

PURPOSE: Retinoids are well known inhibitors of estrogen-dependent breast cancer cell growth and differentiation. alpha2beta1 integrins are involved in the normal growth and differentiation of breast cells, they also take part in many pathological processes including malignancies. The aim of the study was to evaluate the effect of estradiol and tamoxifen on the inhibitory action of retinoids on the proliferation of MCF-7 breast cancer cells and alpha2beta1 integrin expression. MATERIALS AND METHODS: Evaluation was based on [3H]thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. Expression of alpha2beta1 was assessed through immunocytochemical analysis. RESULTS: Treatment of cancer cells with the examined compounds and tamoxifen (10 microM) revealed that only 13-cis retinoic acid (13-cis RA) and all-trans retinoic acid (ATRA) (10(-5) M) decreased cells proliferation compared to the tamoxifen group (30.84%+/-3.32, p<0.01 and 31.05%+/-4.67, p<0.01, respectively). The lowest fraction of PCNA positive cells was also observed after the simultaneous addition ATRA (10(-5) M) and tamoxifen (10 microM) (30.75%+/-0.95, p<0.01, compared to the tamoxifen group). Our results showed that the decrease of alpha2beta1 integrin expression by 13-cis RA (10(-5) M, 49.6+/-3.25%) and ATRA (10-9 M, 15.0%+/-5.0) was augmented by tamoxifen and to a lesser extent by estradiol, particularly in the case of ATRA at 10(-7) or 10(-9) M. CONCLUSIONS: This data suggest that tamoxifen augments the inhibitory effect of retinoids on proliferation and alpha2beta1 integrin expression in MCF-7 cells.

Prenatal evaluation of a fetus with trisomy 18 and additional balanced de novo Rob(13;14).

The main aim of this work is to present unusual case with full trisomy 18 and additional robertsonian translocation- Rob (13;14) detected through abnormalities found in prenatal ultrasound examination. A 26 years-old pregnant women with no family history of any reproductive failure underwent level II ultrasound screening in 19 weeks of gestation. Polyhydramnios, intrauterine growth retardation, hydrocephalus, enlarged lateral ventricles, club foot and cardiac defect were found. Amniocentesis was indicated considering the high likelihood of a chromosomal aberration. Abnormal karyotype was detected 46, XY, der(13;14)(q10;q10), +18. Karyotypes of parents were normal, what confirmed de novo origin of this aberration. Pregnancy was terminated. In postnatal examination fetus demonstrated intrauterine groth retardation and a lot of dysmorphic features characteristic for trisomy 18: microcephaly, prominent occiput, very low set and posteriorly rotated ears, hypertelorism, small mouth, small recessed mandible, a high narrow palate, broad nasal bridge, low-set ears, preauricilar skin appendage, clenched fingers clinodactyly of Vth fingers and club foot. In conclusion it is worth to say that our described fetus demonstrated rather typical for trisomy 18 ultrasonographic features. Balanced Rob (13;14) gives no phenotypic expression. Possible interchromosomal effect in complex chromosomal aberration formation such as Rob (13;14) with trisomy 18 was discussed.

Does variability in carotenoid composition and concentration in tissues of the breast and reproductive tract in women depend on type of lesion?

PURPOSE: Vitamin A takes part in many physiological and pathological processes in women's reproductive organs. The study objective was to compare the carotenoid content in benign and malignant lesions of the breast, ovary and uterus, and to demonstrate quantitative and qualitative similarities or differences between the study groups. MATERIALS AND METHODS: Materials for analysis were physiological and pathological tissues of breast, ovary and uterus. The carotenoid pigments were isolated using column chromatography (CC), thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). RESULTS: Sixteen carotenoids were identified in the study material, including those belonging to the provitamin A group. The most common were: beta-carotene, beta-cryptoxanthin, lutein, mutatoxanthin, violaxanthin, lutein epoxide and zeaxanthin. All the tissues subjected to analysis contained beta-carotene, 98% of the tissues had beta-cryptoxanthin, whereas alpha-carotene was detected in about 50% of breast tissue. No differences in carotenoid concentration were found between benign and malignant lesions in the examined tissues, apart from hydroxyechinenone, canthaxanthin, astaxanthin, lutein epoxide, antheraxanthin and neoxanthin. Similarly, no differences in concentration of the provitamin A carotenoids (alpha-carotene, beta-carotene, beta-cryptoxanthin, and echinenone) were found between benign and malignant lesions except hydroxyechinenone. The highest total content of carotenoids and the biggest spectrum of predominant carotenoids were found in the breast. Only in tissues of malignant lesions of the uterus, we observed statistically higher total content of carotenoids compared to remaining samples from the uterus (p<0.001) and more frequent isolation of some carotenoids (compared to benign lesions). CONCLUSION: The results of our study confirmed the presence of a high diversity of carotenoids in the physiologic, benign and malignant tissues of the breast and the reproductive tract in women. The differences observed among the frequency of isolation of some carotenoids do not allow to make straightforward conclusions. The frequent isolation of provitamin A carotenoids in the examined material and the lack of their occurrence as major carotenoids may be connected with using them in the cellular biological processes. However, this requires further investigation.

Apelin, a novel adipocytokine, in relation to endothelial function and inflammation in kidney allograft recipients.

Apelin, a newly discovered adipocytokine produced by white adipose tissue, is also expressed in kidney and heart. It has been reported that apelin is related to echocardiographic features in hemodialyzed patients. Cardiovascular disease is a major contributor to the mortality and morbidity among patients with chronic renal failure as well as kidney allograft recipients. The aim of this study was to assess the association between apelin and coronary artery disease (CAD) among kidney allograft recipients. We investigated plasma apelin levels in 100 clinically stable, kidney allograft recipients with versus without CAD. We also assessed markers of endothelial cell injury-von Villebrand factor (vWF), thrombomodulin, intracellular adhesion molecule (ICAM), and CD146; markers of inflammation-high-sensitivity-reactive protein (hsCRP); other hemostatic parameters-tissue plasminogen activator (tPA) and its inhibitor (PAI-1); as well as other adipocytokines-adiponectin and resistin-using commercially available kits. Markers of endothelial dysfunction and inflammation were significantly elevated among patients with CAD levels, as well as with CAD or diabetes, compared with those without CAD. Apelin was significantly lower among patients with CAD, but higher in diabetic patients. Apelin content was similar in hypertensive versus normotensive kidney allograft recipients. We observed significant correlations between apelin and ICAM, resistin, adiponectin, calcium, phosphate, alanine and aspartate aminotransferase levels, with CAD or diabetes. Upon multiple regression analysis as well as CAD, adiponectin, and ICAM were predictors of apelin. Apelin was significantly reduced in kidney allograft recipients with CAD; its level was predicted by the presence of CAD, endothelial damage, or inflammation. Apelin and other adipocytokines may be associated with inflammation and its clinical consequences.

Hyaluronic acid counteracts interleukin-1-induced inhibition of collagen biosynthesis in cultured human chondrocytes.

Although, hyaluronic acid (HA) is used in the treatment of osteoarthritis for 30 years, the mechanism of its protective action on collagen metabolism disturbances in tissues during inflammation is not known. Therefore, the present study was undertaken to evaluate the mechanism of IL-1beta action (inductor of experimental inflammation) on deregulation of collagen biosynthesis in cultured human chondrocytes and the effect of HA on the process. It has been found that IL-1beta strongly induced inhibition of collagen biosynthesis, while HA counteracted the process. The mechanism of this phenomenon was found at both transcriptional and post-transcriptional level. IL-1 was found to down regulate the expression of mRNA for type II collagen and to inhibit prolidase activity, an enzyme that plays an important role in collagen biosynthesis at post-translational level. HA was shown to counteract the IL-1beta-dependent inhibition of both processes. During experimental inflammation of chondrocytes cultured in 0.1% FBS there was no differences in the expression of beta(1)-integrin independently of cell number and the presence of HA in growth medium. In chondrocytes cultured in 5% FBS, IL-1beta up-regulated the expression of beta(1)-integrin receptor while HA abolished the effect. The data suggest that HA-dependent up-regulation of collagen biosynthesis in IL-1beta-treated chondrocytes may involve stimulation of prolidase activity in serum "starved" cells and may also originate at the transcriptional level in the cells cultured in standard conditions.

Tartrate-resistant acid phosphatase 5b and its correlations with other markers of bone metabolism in kidney transplant recipients and dialyzed patients.

PURPOSE: Renal osteodystrophy is a common complication of chronic renal failure and renal replacement therapy. Successful kidney transplantation reverses many of these abnormalities, however, the improvement is often incomplete. The osteoclast specific 5b isoform of tartrate-resistant acid phosphatase (TRAP) 5b has recently been proposed a specific and sensitive marker of bone resorption. The aim of the study was to assess correlations of TRAP 5b with markers of bone resorption and formation in kidney transplant recipients, hemodialyzed and peritoneally dialyzed patients and healthy volunteers. MATERIAL AND METHODS: We assessed PTH, markers of bone formation-alkaline phosphatase and its bone isoform, osteocalcin, markers of bone resorption--procollagen type I carboxy-terminal extension peptide, procollagen type I cross-linked carboxy-terminal telopeptide, serum CrossLaps-Ctx, beta2-microglobulin and urinary deoxypyridynoline (DPD), expressed as DPD/creatinine ratio. (BMD) bone mineral density measurements were determined for femoral neck and lumbar spine (L2-L4) using DEXA. RESULTS: In dialyzed patients markers of bone formation and resorption were significantly higher than in healthy volunteers, whereas in kidney transplant recipients these disturbances were less pronounced. TRAP 5b correlated positively with age and mainly with markers of bone resorption in kidney transplant recipients, dialyzed patients and healthy volunteers. TRAP 5b did not correlate with BMD in any groups studied. CONCLUSION: Since TRAP 5b correlated mainly with markers of bone resorption, it may serve as a new additional marker of bone resorption in the assessment of renal osteodystrophy.

Relationship between insulin-like growth factors (IGF-I and IGF-II), IGF-binding proteins (IGFBP-3, IGFBP-2), leptin and anthropometric parameters (height, body mass index) during antileukaemic treatment in children.

PURPOSE: The aim of the study was to estimate the anthropometric parameters and their relationship to serum levels of IGF-I, IGF-II, IGFBP-3, IGFBP-2 and leptin before and during intensive antineoplastic treatment for acute lymphoblastic leukaemia in children. MATERIAL AND METHODS: In 46 children in median age 6.6 years (range from 1.6 to 16) we evaluated at the time of diagnosis, after protocol I and after intensive treatment, height, body mass index (BMI) and IGF-I, IGF-II, IGFBP-3, IGFBP-2 and leptin. RESULTS: Height SDS lowered in successive points of analysis whereas BMI SDS rose after protocol II. IGF-I SDS was low and similar at each point, IGF-II SDS and IGFBP-3 SDS values augmented progressively and IGFBP-2 SDS was significantly elevated before treatment and lowered (but not normalized) during the therapy. Leptin SDS was elevated, especially after protocol I. CONCLUSION: Leukaemia and its treatment affect directly growth factors, its binding proteins and leptin production leading to growth retardation and overweight.

Correlations between leptin, body composition, bone mineral density, and bone metabolism in kidney transplant recipients.

INTRODUCTION: In kidney transplant recipients leptin levels are often elevated and bone mineral density (BMD) decreased. However, to date there are no about correlations between leptin and BMD in this population. It has been suggested that leptin is a predictor of BMD in postmenopausal women. Moreover, leptin acts as a marker of fat stores. We examined the relationships between leptinemia, some markers of nutritional status, BMD, and bone metabolism in kidney transplant recipients. We also assessed whether leptin was a significant and independent predictor of BMD in this population. METHODS: BMD and fat content (global, percentage, trunk) were measured using dual-energy X-ray absorptiometry in 27 kidney allograft recipients. Markers of bone turnover and leptin were studied using commercially available kits. RESULTS: Leptin correlated with the percentage of body fat, trunk fat, lean body mass, serum creatinine, and urea. Insulin growth factor binding protein 1 was negatively related to waist-hip ratio and global and trunk fat, whereas BMD of the lumbar spine was correlated with the daily dose of prednisone, azathioprine, cyclosporine trough levels, serum calcium, as well as osteoprotegerin level. CONCLUSIONS: Leptin levels are associated with graft function and body fat in kidney allograft recipients. Leptin is not related to nutritional status, BMD, or bone metabolism in kidney allograft recipients, but is associated with the current dosage of immunosuppressants and the serum calcium.

Expression of glucose transporter GLUT-1 and estrogen receptors ER-alpha and ER-beta in human breast cancer.

We attempted to describe a GLUT-1 expression in breast cancer and characterize correlation between GLUT-1 and ERs alpha and beta expression as well as correlate this with clinicopathologic features. Sixty-nine patients were involved in the study. GLUT-1, ER-alpha and ER-beta immunocytochemistry was performed using the streptavidin- biotin method. Thirty-seven (53.6%) out of total 69 were GLUT-1 positive. Of GLUT- 1 positive 45.3% were ER-alpha-positive, whereas 81.3% of ER-alpha-negative were GLUT-1 positive. Statistically significant correlation was observed between GLUT-1 and ER-alpha expression status but neither between GLUT-1 and ER-beta nor with clinicopathologic features. No statistically significant correlation was found between expression level (expressed as immunocytoreactive score) of GLUT-1, ER-alpha and ER-beta. Since most of ER-alpha-negative (81.3%) were GLUT-1 positive and significant correlation exists between the two receptors it is reasonable to assume that some functional relation might exists between the expression of two receptors.

Comparison of cartilage self repairs and repairs with costal and articular chondrocyte transplantation in treatment of cartilage defects in rats.

In our experiment, we tried to assess the potential of repair of full-thickness defects in articular cartilages of rabbit femurs. An artificially made, full-thickness defect in the rabbit's femoral patellar groove was created. The defects were divided into six groups. The reparative tissue was evaluated by macroscopic, histological, and immunohistochemical examinations. The reparative tissues in defects with transplanted chondrocytes, had mostly a hyaline-like cartilage appearance and were firmly attached to the surrounding normal cartilage. Only in the control group with periosteal flap and broken subchondral plate, there were signs of partial repair. Self repair of rabbit articular cartilage is very limited. Transplantation of chondrocytes, costal and articular, without differences between groups, is a very potential treatment, producing hyaline-like repair tissue with good histological results.

Association of collagen type I alpha1 gene polymorphism with bone density in survivors of childhood cancer--preliminary report.

A candidate gene, involved in the regulation of bone mass is the COLIA1 gene encoding type I collagen, the major protein of bone matrix. The disease per se, the age of its onset and treatment options might exert an impact on bone mineralization in survivors of childhood malignancy. We examined possible allelic influences of COLIA1 gene polymorphism on BMI, BMD spine and total body in 41 survivors (15 girls) of childhood cancer (the mean age 8.9 years). Genotype distribution was 33 (80.5%) SS and 8 (19.5%) Ss. There were no differences in SDS BMD and SDS BMI between patients with SS and Ss genotype. A tendency towards lower SDS values of BMD spine and BMI was observed (not significant). In conclusion, our preliminary observations suggest that COLIA1 genotype may affect bone accrual in a population treated for childhood cancer. Further investigations in a greater population are needed.

Male gonadal function before and after chemotherapy in prepubertal boys.

UNLABELLED: We evaluated gonadal function in twenty prepubertal boys (6.87 +/- 3.84 years old) at diagnosis and 2.5 +/- 1.6 years after treatment for acute lymphoblastic leukaemia (ALL). We measured serum levels of inhibin B (RIA method), testosterone, FSH, LH (immunoenzymatic methods) and compared the results with controls (31 healthy boys in prepubertal stage). RESULTS: Serum inhibin B levels were lower at diagnosis (55.81 ng/ml +/- 33.74), comparing to respective values in the controls (105.89 ng/ml +/- 46.64), p<0.0002. The values of inhibin B slightly augmented after 2.5 +/- 1.6 years from chemotherapy (79.47 ng/ml +/- 47.39), p<0.06 but remained lower than those in the controls (p<0.07). We did not find any differences in FSH, LH or testosterone values, before and after chemotherapy, comparing to respective values in the controls. In conclusion, haematological malignancy and its treatment influence gonadal function before puberty with a possibility of recovery. During this period, inhibin B could be used as a sensitive indicator of testicular function.

Effects of cathepsin D inhibitor from Vicia sativa L. seed hulls on human skin fibroblasts and breast cancer cells (in vitro studies).

Cathepsin D is a lysosomal protease which plays an important role in cancer invasion and metastasis. There are known inhibitors of that enzyme, such as pepstatin and potato inhibitor. In this study, we examined effects of the cathepsin D inhibitor from Vicia sativa L. seed hulls on cell cultures of human skin fibroblasts and breast cancer cells. There is no effect of the D-cathepsin inhibitor from Vicia sativa L. seed hulls on the proliferative activity of either human skin fibroblasts or breast cancer cells, measured by the [3H] thymidine incorporation assay.