Towards understanding leydigioma: do G protein-coupled estrogen receptor and peroxisome proliferator-activated receptor regulate lipid metabolism and steroidogenesis in Leydig cell tumors?

Leydig cell tumors (LCT) are the most common type of testicular stromal tumor. Herein, we investigate the G protein-coupled estrogen receptor (GPER) and peroxisome proliferator-activated receptor (PPAR) implication in regulation of lipid homeostasis including the expression of steroidogenesis-controlling molecules in clinical specimens of LCTs and tumor Leydig cells (MA-10). We showed the general structure and morphology of LCTs by scanning electron and light microscopy. In LCTs, mRNA and protein analyses revealed increased expression of GPER and decreased expression of PPARα, ß, and γ. Concomitantly, changes in expression pattern of the lutropin receptor (LHR), protein kinase A (PKA), perilipin (PLIN), hormone sensitive lipase (HSL), steroidogenic acute regulatory protein (StAR), translocator protein (TSPO), HMG-CoA synthase, and reductase (HMGCS, HMGCR) were observed. Using MA-10 cells treated with GPER and PPAR antagonists (alone and in combination), we demonstrated GPER-PPAR-mediated control of estradiol secretion via GPER-PPARα and cyclic guanosine monophosphate (cGMP) concentration via GPER-PPARγ. It is assumed that GPER and PPAR can crosstalk, and this can be altered in LCT, resulting in a perturbed lipid balance and steroidogenesis. In LCTs, the phosphatidylinositol-3-kinase (PI3K)-Akt-mTOR pathway was disturbed. Thus, PI3K-Akt-mTOR with cGMP can play a role in LCT outcome and biology including lipid metabolism.

Leydig cell tumorigenesis - implication of G-protein coupled membrane estrogen receptor, peroxisome proliferator-activated receptor and xenoestrogen exposure. In vivo and in vitro appraisal.

The etiology and molecular characteristics of Leydig cell tumor (LCT) are scarcely known. From the research data stems that estrogen can be implicated in LCT induction and development, however it is not investigated in detail. Considering the above, herein we analyzed the relation between G-protein coupled membrane estrogen receptor, peroxisome proliferator-activated receptor and insulin-like family peptides (insulin-like 3 peptide; INSL3 and relaxin; RLN) expressions as well as estrogen level with impact of xenoestrogen (bisphenol A; BPA, tetrabromobisphenol A; TBBPA, and tetrachlorobisphenol A; TCBPA). While in our previous studies altered GPER-PPAR partnership was found in human LCT being a possible cause and/or additionally effecting on LCT development, here mouse testes with experimentally induced LCT and mouse tumor Leydig cell (MA-10) treated with BPA chemicals were examined. We revealed either diverse changes in expression or co-expression of GPER and PPAR in mouse LCT as well as in MA-10 cells after BPA analogues when compared to human LCT. Relationships between expression of INSL3, RLN, including co-expression, and estrogen level in human LCT, mouse LCT and MA-10 cells xenoestrogen-treated were found. Moreover, involvement of PI3K-Akt-mTOR pathway or only mTOR in the interactions of examined receptors and hormones was showed. Taken together, species, cell of origin, experimental system used and type of used chemical differences may result in diverse molecular characteristics of LCT. Estrogen/xenoestrogen may play a role in tumor Leydig cell proliferation and biochemical nature but this issue requires further studies. Experimentally-induced LCT in mouse testis and MA-10 cells after BPA exposure seem to be additional models for understanding some aspects of human LCT biology.

Less advanced testicular dysgenesis is associated by a higher prevalence of germ cell neoplasia.

There is a theory that the more evident clinical signs of testicular dysgenesis, the more frequent the neoplastic lesions are. The aim of this study was to relate the incidence of testicular germ cell neoplastic lesions (overt germ cell tumours--GCT or testicular carcinoma in situ) to the intensity of testicular organogenesis disturbances (dysgenesis). Biopsies were taken from 154 testes of the following patients: 23 patients with GCT in the contralateral gonad (CGCT), 41 patients with undescended testes operated in childhood (UDT), 90 with azoo-/oligozoospermia (A/O) diagnosed because of infertility. Assessment of seminiferous epithelium, number of Leydig cells, areal fraction of intertubular space (IS), morphometric analysis of seminiferous tubules diameter and thickness of tubular wall were performed. Monoclonal antibodies against placental like alkaline phosphatase and cytokeratin 18 were applied. Germ cell neoplastic lesions were detected in 7.1% of testes and were associated with disturbed spermatogenesis. Among testes with disturbed spermatogenesis they were found the most frequently in CGCT (22.2% vs. 11.1% in UDT and 3.8% in A/O), where spermatogenesis had the highest score (5.7 +/- 3.8 points vs. 4.2 +/- 2.7 in UDT and 4.6 +/- 2.9 in A/O). In CGCT, signs of testicular dysgenesis were less advanced: the highest tubular diameter was 164.4 +/- 32.3 microm vs. 163.5 +/- 28.6 in UDT and 161.4 +/- 31.5 in A/O, the lowest thickness of tubular wall was 8.9 +/- 3.2 microm vs. 10.2 +/- 3.6 in UDT and 10.2 +/- 3.2 in A/O, lowest IS was 36.9 +/- 14.9% vs. 47.9 +/- 18.0 in UDT and 46.5 +/- 18.5 in A/O, and the lowest percentage of tubules with immature Sertoli cells was 0.1 +/- 0.4% vs. 4.9 +/- 7.0 in UDT and 5.2 +/- 9.7 in A/O. Results indicate that neoplastic lesions appear only in testes with disturbed spermatogenesis. Worse condition of spermatogenesis is associated by the presence of other dysgenetic features, but neoplastic lesions appear more frequently in testes with the less advanced features of testicular dysgenesis.

[Identification of mutation and polymorphic changes in the CFTR gene of patients with obstructive azoospermia]. / Identyfikacja mutacji i zmian polimorficznych w genie CFTR u pacjentów z azoospermia obstrukcyjna.

Obstructive azoospermia is one of the symptoms of congenital bilateral absence of the vas deferens (CBAVD)--disease which is suggested as primarily genital form of cystic fibrosis. CBAVD is a result of mutations and polymorphisms in CFTR gene. We studied 9 the most common mutations and identified 3 mutations (delta F508, R117H, G542X). The study showed that: 37.1% of CBAVD patients (13 out of 35 examined patients) carried mutations in a single or both copies of the gene. We also found the increased frequency of IVS8-5T allele in this group. No increased frequency of mutations in CFTR gene was found in group of men with incorrect spermatogenesis.

Further delineation of the classical Smith-Lemli-Opitz syndrome phenotype at different patient ages: clinical and biochemical studies.

We present here extensive clinical and biochemical data on thirteen SLOS (type I) patients with proven defect in cholesterol biosynthesis for further delineation of the classical SLOS phenotype at different patient ages.

[Congenital malformations of the genitourinary tract in children with Smith-Lemli-Opitz syndrome]. / Wady wrodzone ukladu moczowo-plciowego u dzieci z zespolem Smitha-Lemlego-Opitza.

Smith-Lemli-Opitz (SLO) syndrome is an autosomal, recessive condition. The essence of SLO is abnormal metabolism of cholesterol which is characterized by decreased level of cholesterol and increased level of 7-dihydrocholesterol in serum. The main clinical features are mental retardation and congenital malformations of various organs and systems--microcephaly, syndactyly, craniofacial anomalies, cleft palate and genitourinary system, mainly external genitalia. We analyzed the group of 12 children (2 girls and 10 boys) treated due to SLO in The Children's Memorial Health Institute since 1982 to 1997. In boys we diagnosed: penile curvature; hypospadia without hypospadias; glandular, penile and scrotal hypospadia, undescended; hydrocele testis; mild unilateral hydronephrosis and in girls we reported moderate hypertrophy of clitoral preputium and stenosis of external urethral meatus. The surgical treatment was planned.

[Needle testicular biopsy in the diagnosis of male infertility]. / Iglowa biopsja jader w diagnostyce nieplodnosci meskiej.

OBJECTIVES: Evaluation of testicular tissue is very important diagnostic procedures in cryptozoospermia and azoospermia. It is verified the patient for treatment (stimulation spermatogenesis), for micromanipulation ICSI or for insemination donor semen procedure. Surgical biopsy is very popular between andrologist and urologist, but needle biopsy seems to be easier and safer. DESIGN: The authors present their experience with needle testicular biopsy during the diagnosis patients with azoospermia or cryptozoospermia and the men with paraplegia after trauma and without ejaculation. MATERIAL AND METHODS: The study group included 63 biopsies in 58 men with azoospermia and severe oligoasthenozoospermia (cryptozoospermia) and 2 with paraplegia. The specimens were taken from 125 testes under general anesthesia (i.v-Diprivan, Propofol, Zeneca and Fentanyl) using biopsy needle from Hepafix B. Braun Melsungen, Germany. All procedures were performed as a day case. RESULTS: In 95% specimens were adequate for histopathological investigations and for planning the treatment. Only one complication (0, 8%) -small haematoma testis was observed. CONCLUSIONS: The needle biopsy of testicular tissue is sufficiency in histopathological examination, safe for patients and easy for urologists. The total cost is much more lower than cost of surgical biopsy.

[Percutaneous sperm retrieval for ICSI procedures in men with obstructive azoospermia: ICSI-PESA and ICSI-TESE micromanipulation: our experience]. / Wlasne doswiadczenia w przezskórnym pozyskiwaniu plemników do zaplodnienia pozaustrojowego ICSI-PESA oraz ICSI-TESE u mezczyzn z azoospermia obstrukcyjna.

OBJECTIVES: The sperm retrieval for ICSI procedures is possibly using following procedures: ejaculate sperm, epididymal sperm obtained by microsurgical aspiration or percutaneous puncture and testicular sperm that are obtained by surgical excision or percutaneous biopsy. Percutaneous techniques seem to be rather simple and effective procedures. DESIGN: The authors present their own experiences with percutaneous sperm retrieval for micromanipulation ICSI from the epididymis (ICSI-PESA) and from the testicular tissue (ICSI-TESE) in men with obstructive azoospermia and with reactive impotence. First time in Poland ICSI-PESA was done on April 11, 1996 in Private Infertility Center "Novum", Warsaw. MATERIAL AND METHODS: From April 1996 to the end of January 1998, 10 ICSI-PESA procedures (in 9 couples) and 8 ICSI-TESE (in 6 couples) were performed. In one case ICSI-PESA was performed in man with psychological inability of masturbation during his wife's IVF protocol. All procedures were performed as the day case urology, under general anesthesia. The fine needles No 6 in PESA or biopsy needle from Hepafix Set B. Braun in TESE were used. The therapy of antibiotic and common analgesic drug was used routinely after puncture. RESULTS: The effectiveness of obtaining sperm for micromanipulation were 100% ICSI-PESA and 75% ICSI-TESE. The pregnancy rate in PESA was 50% and 5 healthy children were born. In TESE only 1 woman (17%) was pregnant, but early spontaneous miscarriage was reported. No surgical and anesthesiological complications were noticed. CONCLUSIONS: Obtaining sperm for micromanipulation ICSI using percutaneous epididymal puncture or testicular tissue needle biopsy seems to be effective and safe for patients with obstructive azoospermia or reactive impotence.

["May we not waste their deaths..." Memories of a participant in the "Sonderaktion Krakau"]. / "Bysmy lch smierci nie zmarnowali..." Ze wspomnien uczestnika "Sonderaktion Krakau".

The Author analyses the "Sonderaktion Krakau" from historical and cultural point of view. Striking Polish science and national intellectual elite with an insidious blow, the invader reached historical experiences. In the second part the Author reconstructs fate of imprisoned professors and quotes many personal observations from the period of his stay in the Sachsenhausen and Dachau Nazi camps.

Kabuki (Niikawa-Kuroki) syndrome associated with immunodeficiency.

We report a case of a 19-year-old male with the cardinal features of the Kabuki syndrome (KS) and, in addition, with severe immunodeficiency. Finding immune deficiency in a KS patient, prompted us to determine whether this association was related to a deletion within the DiGeorge chromosomal region. Fluorescence in situ hybridization (FISH) with the Oncor probe N25(D22S75) revealed no deletion of 22q11.2 in the patient.