Friend erythroleukemia cells induce angiogenesis in chick embryo chorioallantoic membrane and in human umbilical vein endothelial cells

The effects of Friend erythroleukemia cells on angiogenesis were studied in chick embryo chorioallantoic membrane assay and in human umbilical vein endothelial cells. In chorioallantoic membrane assay, the conditioned medium of Friend cells stimulated in vivo angiogenesis to an extent comparable to that observed with Prostaglandin El, used as positive control. Prostaglandin El added to conditioned medium of Friend cells did not further increase angiogenesis. Conditioned medium of Friend erythroleukemia cells also stimulated proliferation of human umbilical vein endothelial cells to an extent comparable to that observed with fetal bovine serum, used as positive control. Conditioned medium and fetal bovine serum together did not affect human umbilical vein endothelial cells proliferation, as compared to that observed when tested separately. These results seem to indicate that Friend erythroleukemia cells produce and secrete factors stimulating angiogenesis. These findings extend and confirm the hypothesis that successful angiogenesis is necessary for development of leukemia

Effects of vitamin D-binding protein-derived macrophage-activating factor on human breast cancer cells

BACKGROUND: Searching for additional therapeutic tools to fight breast cancer, we investigated the effects of vitamin D-binding protein-derived macrophage activating factor (DBP-MAF, also known as GcMAF) on a human breast cancer cell line (MCF-7). MATERIALS AND METHODS: The effects of DBP-MAF on proliferation, morphology, vimentin expression and angiogenesis were studied by cell proliferation assay, phase-contrast microscopy, immunohistochemistry and western blotting, and chorioallantoic membrane (CAM) assay. RESULTS: DBP-MAF inhibited human breast cancer cell proliferation and cancer cell-stimulated angiogenesis. MCF-7 cells treated with DBP-MAF predominantly grew in monolayer and appeared to be well adherent to each other and to the well surface. Exposure to DBP-MAF significantly reduced vimentin expression, indicating a reversal of the epithelial/mesenchymal transition, a hallmark of human breast cancer progression. CONCLUSION: These results are consistent with the hypothesis that the known anticancer efficacy of DBP-MAF can be ascribed to different biological properties of the molecule that include inhibition of tumour-induced angiogenesis and direct inhibition of cancer cell proliferation, migration and metastatic potential

Role of angiotensin-converting enzyme and vitamin D receptor gene polymorphisms in cancer anorexia-cachexia syndrome

The ubiquitin-proteasome pathway is a crucial connection between aberrant immune system activation, systemic inflammation and Cancer Anorexia-Cachexia Syndrome (CACS), a syndrome that culminates in hyper-activation of the ubiquitin-proteasome pathway. Angiotensin directly up-regulates this pathway, while vitamin D down-regulates it indirectly through the insulin-like growth factor-1 pathway. We investigated the genetic predisposition towards CACS in a cancer population, examining Insertion/Deletion (I/D) polymorphism of angiotensin-converting enzyme gene and FokI and BsmI polymorphisms of vitamin D receptor gene. Sixty-two cancer patients were recruited and divided into three groups: primary cachectic (C1, n = 14; dysmetabolic body weight loss ≥5% in 6 months); secondary cachectic (C2, n = 34; similar weight loss, mechanic or iatrogenic origin); and non-cachectic (NC, n = 16). C2+NC were merged in the control group. The three groups showed significant differences in average prognostic inflammatory nutritional index (C1: 26.4±23.4; C2: 5.4±5.6; NC: 0.37±0.5), C-reactive protein serum levels (C1: 6.6±2.1; C2: 2.4±2.2; NC: 1.0±2.0 mg/dL), albumin serum levels (C1: 3.1±0.6; C2: 3.5±0.4; NC 3.7±0.6 g/dL), weight loss (C1: 22±8; C2: 15±6.7; NC 5±6%) and life expectancy (C1: 6.4±3.3; C2: 25±28; NC: 45±25 months). However, none of the chosen polymorphisms showed any statistically significant correlation with CACS. The complexity of the changes of the immune system in the chronic inflammation state associated with CACS is far greater than expected and further studies are required to identify genetic independent markers of progression toward CACS.

The pharmacological perturbation of brain zinc impairs BDNF-related signaling and the cognitive performances of young mice.

Zinc (Zn2+) is a pleiotropic modulator of the neuronal and brain activity. The disruption of intraneuronal Zn2+ levels triggers neurotoxic processes and affects neuronal functioning. In this study, we investigated how the pharmacological modulation of brain Zn2+ affects synaptic plasticity and cognition in wild-type mice. To manipulate brain Zn2+ levels, we employed the Zn2+ (and copper) chelator 5-chloro-7-iodo-8-hydroxyquinoline (clioquinol, CQ). CQ was administered for two weeks to 2.5-month-old (m.o.) mice, and effects studied on BDNF-related signaling, metalloproteinase activity as well as learning and memory performances. CQ treatment was found to negatively affect short- and long-term memory performances. The CQ-driven perturbation of brain Zn2+ was found to reduce levels of BDNF, synaptic plasticity-related proteins and dendritic spine density in vivo. Our study highlights the importance of choosing "when", "where", and "how much" in the modulation of brain Zn2+ levels. Our findings confirm the importance of targeting Zn2+ as a therapeutic approach against neurodegenerative conditions but, at the same time, underscore the potential drawbacks of reducing brain Zn2+ availability upon the early stages of development

Effects of Cadmium and vitamin D binding protein-derived macrophage activating factor (DBP-MAF) in human breast cancer cells

We previously demonstrated that chronic exposure of the human breast cancer cell line MCF-7 to non-cytotoxic concentrations of Cadmium reduced viability and angiogenic potential of this cell line. In order to better understand these effects, cells, after Cadmium exposure, were treated with vitamin D binding protein-derived macrophage activating factor (DBP-MAF). DBP-MAF is a potent macrophage-activating factor derived from vitamin D binding protein, a polymorphic serum glycoprotein with multiple functions also known as a group specific component or Gc protein. Besides stimulating macrophages, DBP-MAF has anti-tumour properties. Our data demonstrate that the decrease of MCF-7 cell viability following Cadmium treatment was completely reversed when DBP-MAF was present in the cell medium. Following this observation, we further investigated the role of DBP-MAF in modulating angiogenesis, morphology and cytoskeleton structure of MCF-7 cell line. As shown by chorioallantoic membrane assay, DBP-MAF inhibited MCF-7 cancer cell-stimulated angiogenesis. Concerning cell morphology (studied by contrast phase light microscopy and after Papanicolaou staining), following DBP-MAF treatment, cell shape and growth pattern were significantly modified. Vimentin expression (studied by immunohistochemistry and Western blot analysis), considered a hallmark of human breast cancer progression, after DBP-MAF treatment, significantly varied. Intermediate filament status changes, consisting in a shift from a keratin-rich to a vimentin-rich network (epithelial-mesenchymal transition), were observed. In conclusion, we demonstrate that the anti-cancer effects of DBP-MAF can be attributed to multiple actions independent of macrophage stimulation such as reversal of Cadmium effects on cell viability, reversal of morphological malignant phenotype and inhibition of cancer cell-stimulated angiogenesis. For these reasons, DBP-MAF might represent an useful tool to control progression and differentiation of human breast cancer

Macrophages of the mucosa-associated lymphoid tissue (MALT) as key elements of the immune response to vitamin D binding protein-macrophage activating factor

Macrophages are key elements of the immune response and vitamin D binding protein-macrophage activating factor (DBP-MAF, also known as GcMAF) has been successfully used in treatment of immunodeficiency (J Med Virol 81:16-26, 2009). Here we report the effects of DBP-MAF on the immune system of HIV/AIDS patients as well as the effects of an original probiotic preparation, putatively containing DBP-MAF. Eight HIV/AIDS patients were treated with 100 ng/week DBP-MAF (from www.gcmaf.eu) i.v. for 15 weeks. During treatment, patients did not assume antiretroviral drugs. Blood monocyte count rose in six patients, indicating a response to DBP-MAF consistent with the effects of DBP-MAF described in Immunol Cell Biol 76:237-44, 1998. Individual response appeared to be associated with vitamin D receptor (VDR) gene polymorphisms (BsmI and FokI). Within the time frame of administration, however, no significant increase in CD4 cell count or decrease in viral load was observed. Therefore, searching for an alternative approach, we tested an original milk-derivative (MAF 3 14®) that contains microorganisms introduced in order to maximize natural DBP-MAF production. We hypothesized that natural DBP-MAF, once ingested, activated the Mucosa-Associated Lymphoid Tissue (MALT) widely diffused in the walls of the entire gastrointestinal tract. In fact, enzymes of certain strains of microorganisms contained in yogurt and kefir are able to convert milk Gc-protein into active DBP-MAF and it is known that kefir modulates the immune response in mice, increasing the phagocytic activity (i.e. activating) of peritoneal and pulmonary macrophages (Immunobiology 211:149-56, 2006). It is also known that probiotic yogurt consumption is associated with an increase of CD4 count among people living with HIV/AIDS (J Clin Gastroenterol 44:e201-5, 2010). Thus, members of the research team consumed 125 ml/day of MAF 3 14® for three weeks. Participants did not assume any drug or supplement and did not modify their usual diet and lifestyle. Blood analyses were performed two weeks before beginning consumption, and after three week consumption. After three week consumption, CD4 count dramatically increased in those of us who started with low CD4 count (subject # 1, before consumption CD4: 372; CD8: 206. After consumption: CD4: 609; CD8: 448), or abnormal CD4/CD8 ratio (subject # 2, before consumption: CD4: 857; CD8: 794. After consumption: CD4: 1279; CD8: 640). Also these effects appeared to be associated with VDR gene polymorphisms

ACE/VDR gene polymorphisms and bioelectrical impedance analysis in predicting athletic performances of Italian Young soccer players.

We evaluated the association between 2 genetic polymorphisms known to be involved in fitness and performance, and anthropometric features, body composition, and athletic performances in young male soccer players with the goal of identifying genetic profiles that can be used to achieve maximal results from training. One hundred twenty-five medium-high-level male soccer players were genotyped for angiotensin-converting enzyme (ACE) I/D, and vitamin D receptor (VDR) FokI gene polymorphisms and scored for anthropometric measurements, body composition, and athletic performance. Body mass index, fat mass, fat-free mass, resistance, reactance, impedance, phase angle (PA), and body cell mass were measured. Athletic performance was evaluated by squat jump, countermovement jump (CMJ), 2-kg medicine ball throw, 10- and 20-m sprint time. We observed that the homozygous ff genotype of the VDR gene was significantly more represented in young soccer players than in a matched sedentary population. Values of reactance and PA were differently distributed in ACE and VDR genotypes with high mean values in subjects with DD (ACE) and FF (VDR) genotypes. No correlation was observed between ACE or VDR genotypes and 2-kg medicine ball throw, 10- and 20-m sprint times. The ID genotype of ACE was associated with the best performances in squat jump and CMJ. Our results suggest that determination of ACE and VDR genotypes might help select those young athletes harboring the most favorable genetic potential to succeed in soccer

Association between vitamin D receptor gene polymorphisms and iron indices in HIV-negative patients with malnutrition-inflammation-cachexia syndrome

Malnutrition-inflammation-cachexia syndrome (MICS) is a frequent complication of end-stage AIDS. Malnutrition, with its associated adverse effects on immunocompetence contributes to the progression of AIDS independently of HIV. Iron indices are considered reliable prognostic factors because of their association with inflammation and malnutrition. Since the active form of vitamin D has immunomodulatory effects, and considering that allelic variants of the vitamin D receptor (VDR) are associated with the rates of progression to AIDS in HIV-positive patients (J Steroid Biochem Mol Biol. 2004 89-90:199-207), here we evaluated the association between VDR polymorphisms and iron indices in HIV-negative patients with MICS. 38 HIV-negative patients treated at the Unit of Clinical and Artificial Nutrition, Misericordia e Dolce Hospital, Prato (Italy) for at least 12 months were studied. The mean age of the patients (22 men and 16 women) included in this study was 65 ± 10 years. Genomic DNA was extracted from peripheral blood leucocytes and amplified by polymerase chain reaction (PCR). PCR products were digested with the respective restriction enzymes in order to identify VDR polymorphisms. Absence or presence of the BsmI, ApaI, TaqI, and FokI restriction sites were denominated B and b, A and a, T and t, F and f respectively. Serum transferrin levels showed significant association with BsmI, ApaI and TaqI polymorphisms, i.e. those polymorphisms that are located in a regulatory site at the 3’ end of the VDR gene and are in linkage disequilibrium. Patients harbouring the BB, AA and tt genotypes showed significantly higher levels of serum transferrin compared with bb, Aa, aa, TT and Tt respectively. These results are consistent with those previously obtained in HIV-positive patients (J Steroid Biochem Mol Biol. 2004 89-90:199-207; J Infect Dis. 2008 197:405-10) and highlight an inverse correlation between vitamin D signalling and AIDS progression to MISC. These results also provide a link between VDR alleles and nutritional markers which are highly predictive variables of MICS. Since MICS is one of the leading causes of mortality in AIDS patients, the determination of VDR polymorphisms could help identifying those AIDS patients with a greater risk of developing MICS, a syndrome that appears to be independent of HIV serostatus or viral load