Efficacy and safety of hormonal therapy with androgens (androgel) in men with erectile dysfunction, partial androgen deficiency of aging male and cardiovascular diseases

Partial androgen deficiency of aging male (PADAM) manifests with sexual dysfunction and is associated with many diseases, primarily, cardiovascular. After the age of 30-40 a testosterone level falls 1-2% a year. The number of men with testosterone deficiency grows from 8% in 40-60-year-olds to 85% at the age over 80 years. Low testosterone correlates with such risk factors of cardiovascular diseases as dyslipidemia, atherosclerosis, low fibrinolysis, insulin resistance and abdominal obesity. Correction of androgenic deficiency can be conducted with the drug androgel which represents a new system of transdermal testosterone delivery. In contrast to vasoactive drugs, androgel affects pathogenetic mechanisms of erectile dysfunction and thus attenuates factors of cardiovascular risk. Androgel is used externally and is more effective than intramuscular and oral analogues. Also, the drug improves lipid spectrum. By activating lipolysis, testosterone reduces the amount of visceral fat thus lowering insulin resistance. A vasodilating effect of androgel positively influences cardiovascular system and penile vessels. The drug acts fast, is effective and safe. Therefore, it can be recommended for correction of erectile dysfunction in patients with old age androgen deficiency and concurrent cardiovascular diseases.     

Hepatic erythropoietin (Ep) production following double partial hepatectomy in the rat

Double partial hepatectomy (hepx) evokes an elevation in serum erythropoietin (Ep) levels in anephric hypoxic animals when compared to non-hypoxic or sham hepx controls. But this Ep response is significantly lower than that found in singly hepx, anephric hypoxic rats. Double hepx also induces numerous cytological changes in the liver. Extravascular accumulation of fat, fibrous scarring, localized necroses, and multiple abscesses, as well as decreased vascularity, occur following the second hepx. A humoral factor was detected in the serum of these animals that is capable of inducing hepatic Ep production when injected into normal rats 18 hours before nephrectomy and hypoxia. This factor, termed hepatopoietin (Hp), was previously demonstrated in the venous serum of singly hepx rats. The serum from animals subjected to double partial hepx is not as potent in inducing Ep production as the serum from singly hepx animals. The discrepancies noted between the single and double hepx groups is attributed to the necrotic cytological changes described above.     

Significance of eosinophil counting in tumor associated tissue eosinophilia (TATE)

Eosinophils are present in large numbers in some squamous cell carcinomas of the oral cavity. Whilst it is proposed that they have an 'immuno-protective' effect, this remains unproven. The contradictory reports may be due to inconsistencies in eosinophil counting. Eighty-one cases of squamous cell carcinoma (SSC) of oral tongue were examined. Two methods of eosinophil counting were performed. In the first method (classical), the eosinophils were counted per 10 HPF. In the second method (our so-called density method), the highest eosinophil density per surface area was counted for each case. The two methods were correlated. Using the classical method a number of fields in cases ranked low, contained more than 10 eosinophils. Likewise, some moderate cases contained more than 100 eosinophils. There is poor correlation between the classical and density counts. Nevertheless, good correlation between the two methods could be achieved if the boundaries of the classical method are modified. Eosinophils invariably appear in clusters. We feel that an assessment of density may well be better than classical counting, and have more relationship with function.     

Early detection of tumor cells by innate immune cells leads to T(reg) recruitment through CCL22 production by tumor cells

In breast carcinomas, patient survival seems to be negatively affected by the recruitment of regulatory T cells (T(reg)) within lymphoid aggregates by CCL22. However, the mechanisms underpinning this process, which may be of broader significance in solid tumors, have yet to be described. In this study, we determined how CCL22 production is controlled in tumor cells. In human breast carcinoma cell lines, CCL22 was secreted at low basal levels that were strongly increased in response to inflammatory signals [TNF-α, IFN-γ, and interleukin (IL)-1β], contrasting with CCL17. Primary breast tumors and CD45(+) infiltrating immune cells appeared to cooperate in driving CCL22 secretion, as shown clearly in cocultures of breast tumor cell lines and peripheral blood mononuclear cells (PBMC) or their supernatants. We determined that monocyte-derived IL-1β and TNF-α are key players as monocyte depletion or neutralization of these cytokines attenuated secretion of CCL22. However, when purified monocytes were used, exogenous human IFN-γ was also required to generate this response suggesting a role for IFN-γ-producing cells within PBMCs. In this setting, we found that human IFN-γ could be replaced by the addition of (i) IL-2 or K562-activated natural killer (NK) cells or (ii) resting NK cells in the presence of anti-MHC class I antibody. Taken together, our results show a dialogue between NK and tumor cells leading to IFN-γ secretion, which in turn associates with monocyte-derived IL-1β and TNF-α to drive production of CCL22 by tumor cells and subsequent recruitment of T(reg). As one validation of this conclusion in primary breast tumors, we showed that NK cells and macrophages tend to colocalize within tumors. In summary, our findings suggest that at early times during tumorigenesis, the detection of tumor cells by innate effectors (monocytes and NK cells) imposes a selection for CCL22 secretion that recruits T(reg) to evade this early antitumor immune response.     

Targeted delivery of RNA-cleaving DNA enzyme (DNAzyme) to tumor tissue by transferrin-modified, cyclodextrin-based particles

Short nucleic acid sequences specific to oncogene targets such as bcl-2, bcr-abl, and c-myc have been shown to exhibit specific anti-cancer activity in vitro through antigene or antisense activity. Efficient in vivo delivery of oligonucleotides remains a major limitation for the therapeutic application of these molecules. We report herein on the preparation of transferrin-modified nanoparticles containing DNAzymes (short catalytic single-stranded DNA molecules) for tumor targeting as well as their biodistribution using various methods of administration in the mouse. Linear, beta-cyclodextrin-based polymers are complexed with DNAyzme molecules to form sub-50 nm particles termed "polyplexes". The surface properties of the cyclodextrin-containing polyplexes are modified by exploiting the ability of the beta-cyclodextrin substructure and adamantane to form inclusion complexes. Accordingly, conjugates of adamantane with poly(ethylene glycol) (PEG) are prepared and combined with the polyplexes. The adamantane form inclusion complexes with the surface cyclodextrins of the polyplexes to provide a sterically stabilizing layer of PEG. The stabilized polyplexes are also modified with transferrin for increasing targeting to tumor cells expressing transferrin receptors. The preparation, characterization, and in vitro application of these nanoparticles are discussed. The transferrin-polyplexes containing fluorescently-labeled DNAzyme molecules are administered to tumor-bearing nude mice and their biodistribution and clearance kinetics are monitored using a fluorescence imaging system. Four methods of administration are studied: intraperitoneal bolus and infusion, intravenous bolus, and subcutaneous injection. DNAzymes packaged in polyplex formulations are concentrated and retained in tumor tissue and other organs, whereas unformulated DNAzyme is eliminated from the body within 24 hours post-injection. Intravenous and intraperitoneal bolus injections result in the highest fluorescent signal (DNAzyme) at the tumor site. Tumor cell uptake is observed with intravenous bolus injection only, and intracellular delivery requires transferrin targeting.     

Regulation of vascular endothelial growth factor (VEGF) production and angiogenesis by tissue Factor (TF) in SGC-7901 gastric cancer cells

Tissue factor (TF), an initiator of the extrinsic coagulation cascade, is expressed in a wide range of cancer cells and plays important roles in cancer progression and metastasis. We demonstrated between TF and vascular endothelial growth factor (VEGF) production differences in four human gastric cell lines. One of these cell lines, SGC-7901, a high TF and VEGF producer, was grown subcutaneously in severe combined immuno-deficient (SCID) mice. The SCID mice generated solid tumors characterized by intense vascularity. In contrast, SGC-7901 cells transfected with antisense TF cDNA generated relatively avascular tumors in SCID mice, as determined by immunohistochemical staining of tumor vascular endothelial cells with anti-VIII factor antibody. To investigate the structure-function relationship between TF and VEGF, the SGC-7901 cell line was transfected with antisense a full-length TF cDNA, a cytoplasmic deletion mutant lacking the distal three serine residues (potential substrates for protein kinase C), or an extracellular domain mutant, which has markedly diminished function for activation of factor X. Cells transfected with the full-length antisense TF sequence produced decreased levels of both TF and VEGF. Transfectants with the extracellular domain mutant produced high levels of VEGF mRNA. However, cells transfected with the cytoplasmic deletion mutant construct produced increased levels of TF, but little or no VEGF. Thus, the cytoplasmic tail of TF may signal VEGF expression in some tumor cells.     

Testosterone regulates cell proliferation in aggressive fibromatosis (desmoid tumour)

Background:

Aggressive fibromatosis (desmoid tumour) is a locally invasive tumour caused by mutations resulting in β-catenin protein stabilisation. Apc1638N mice are predisposed to developing aggressive fibromatosis tumours, and male mice develop greater numbers of tumours than female mice, suggesting a role for androgens in this tumour type.

Methods:

Human aggressive fibromatosis tumours were examined for the expression of the androgen receptor, and primary human tumour cell cultures were treated with testosterone. Orchidectomised Apc1638N mice were investigated for the development of tumours, and were treated with testosterone to study the effect of tumour formation and the level of β-catenin.

Results:

Androgen receptors are universally expressed in human aggressive fibromatosis tumours. Testosterone increased the proliferation rate and β-catenin protein level in a dose-dependent manner in human aggressive fibromatosis tumours. Orchiectomy reduced the number and size of tumours that formed in male Apc1638N mice to a similar level as observed in female mice. Testosterone treatment increased the number of tumours that formed in orchidectomised male mice, and resulted in a marked increase in β-catenin protein levels.

Conclusion:

Testosterone regulates β-catenin protein level and proliferation rate in this mesenchymal tumour. This work identifies the therapeutic use of testosterone blockade in aggressive fibromatosis as an area for further investigation.     

Partial androgen deficiency in aging male (Padam syndrome): terminology and current approaches to choice of replacement hormonal therapy

The authors propose a novel approach to treatment of PADAM syndrome based on stimulation of synthesis of endogenic testosterone. A total of 150 patients with PADAM were examined in the Endocrinological Research Center. The examination included physical and andrological examinations with digital rectal investigation of the prostate, filling-in questionnaires, hormonal and biochemical tests. In view of positive test with chorionic gonadotropin (CG), therapy of choice was stimulating therapy with CG. The dose was adjusted individually - from 1000 to 3000 units once in 4 days (1000 units dose was given to 20% patients, 2000 - to 70% and 3000 - to 10% patients). Efficacy did not reduce in long-term treatment. The authors make the following conclusion: choice of PADAM therapy should be individual and based on the age of the patient, body mass index, the necessity to retain spermatogenesis, packed cell volume, associated diseases. Secondary nature of PADAM and positive test for CG allows usage of CG as a perspective therapy in prevention and treatment of age-related lowering of testosterone.     

Paclitaxel (TAXOL(R)) concentrations in brain tumor tissue

BACKGROUND: Paclitaxel (Taxol^®) is a novel chemotherapeutic agent, activeagainst a variety of tumors. It is not known whether the drugpenetrates brain tumor tissue. PATIENTS AND METHODS: Three patients with a recurrent glioma received paclitaxel (175mg/m²) in a 3-hour i.v. infusion prior to surgery. Paclitaxelconcentrations were measured in the tumor tissue, cerebrospinalfluid, cyst fluid, plasma and, in one patient, normal braintissue. RESULTS: Tumor tissue concentrations were in the thera- peutic rangein all three patients. Brain tissue concentration, however,was below the detection limit of the trial. CONCLUSIONS: These findings suggest that paclitaxel may have a place in braintumor therapy. The low concentration in normal brain tissue,as observed in one patient, may suggest, however, that the drugdoes not cross the intact blood-brain barrier. brain tumor, chemotherapy, glioma, paclitaxel, tissue concentrations     

Sheep bronchoalveolar carcinoma: tissue associated protein complex (TAPC) in normal lung tissue and in the tumor differed quantitatively

Sheep lungs experimentally and naturally affected by bronchoalveolar carcinoma were washed out exhaustively of soluble components by phosphate-buffered saline, pH 7.4 (PBS), followed by glycine buffer, pH 2.8 (GB), and then again by 1M KCl followed by PBS. The tissue matrix (TM) of the tumor-free region and the tumor-affected tissue were analysed separately by sodium dodecyl sulfate (SDS) polyacrylamide electrophoresis. Normal lung tissues obtained from normal sheep served as controls. Several protein fractions and fragments, identified in both the normal and the tumorous lung, have the molecular weight (MW) of 130,000-228,000, as compared with the major soluble tissue associated protein having MW of 70,000. Coomassie blue staining used in the SDS polyacrylamide system and alkaline phosphatase immunoreaction used in the Enzyme Linked Immunosorbant Assay (ELISA) showed tenfold increased concentration of the TAPC in the TM of the tumor tissue and in the blood of tumor-affected animals, respectively. Total concentration of the TAPC in the serum of tumor-affected animals was higher than in the normal. Immunofluorescent antibody test (IAT) detected the TAPC in the cytoplasm of tumor as well as in normal lung cells, and the study suggested that the TAPC reaches the peripheral blood during tissue destruction occurring at the tumor site, as observed by light and electron microscopy (LM and EM). The concentration of each of the TAPC fractions was higher in the tumor-affected sheep lung as compared with normal sheep lung. Antibodies prepared against the TAPC fractions were toxic to sheep lung cells in tissue culture. Tumor cells were more susceptible.     

Cultured oncogenic osteomalacia tumor cells produce a factor(s) that inhibits osteocalcin production by osteoblastic cells

A 34-year-old patient was diagnosed with oncogenic osteomalacia associated with hypophosphatemia, low levels of serum 1,25-dihydroxyviamin D [1,25(OH)(2)D], and osteocalcin (OC). Resection of the tumor normalized these blood abnormalities. While such tumors produce a humoral factor(s) that affects phosphate reabsorption by the proximal renal tubules, the direct action of such factor(s) on osteoblast function has not been examined previously. We investigated the effect of conditioned medium of cultured osteomalacia tumor cells on OC production by human osteoblastic cell line, MG-63. The conditiond medium inhibited OC production induced by 1,25(OH)(2)D-3. Our results indicate that the humoral factor(s) produced by the tumor has direct effect on osteoblasts and may contribute to development of the characteristic syndrome.