Anti-spermatogenic activities of Taraxacum officinale whole plant and leaves aqueous extracts.

Taraxacum officinale has been used in Jordan folk medicine to treat male infertility. A recent study has proved a contradictory effect of the whole plant aqueous extract. The aim of the current study was to determine if the leaves of T. officinale have similar anti-fertility activities, and whether this effect is mediated through the regulation of spermatogonial stem cells (SSCs). Fifty adult male rats were divided into five groups. Two groups were gavaged with 1/10 of LD50 of T. officinale whole plant (1.06 g kg(-1) body weight) or leaves (2.30 g kg(-1) body weight) aqueous extract; while two groups were gavaged with 1/20 of LD50 of T. officinale whole plant (2.13 g kg(-1)) or leaves (4.60 g kg(-1)) extract. The control group received distilled water. Oral administration of T. officinale (whole plant and leaves aqueous extract) caused a significant decrease in testis and seminal vesicle weight, a reduction in serum testosterone concentration, impaired sperm parameters, and a decrease in pregnancy parameters. Testicular histology of treated rats showed structural changes such as hypoplasia of germ cells, reduction in the thickness of germinal epithelium, arrest of spermatogenesis at spermatid stage (late maturation arrest) and reduction in the number of Leydig cells. Gene expression levels of two SSCs markers (GFRα1 and CSF1) responsible for self-renewal were relatively counter-balanced. In conclusion, T. officinale whole plant and leaves aqueous extracts changed the gene expression of two SSCs markers leading to the imbalance between spermatogonia self-renewal and differentiation causing late maturation arrest.

2-deoxy-D-Glucose Synergizes with Doxorubicin or L-Buthionine Sulfoximine to Reduce Adhesion and Migration of Breast Cancer Cells.

BACKGROUND: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. MATERIALS AND METHODS: T47D human breast cancer cells were treated with 2-deoxy- D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescent- phalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. RESULTS: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. CONCLUSIONS: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.

Feasibility of collecting umbilical cord blood in jordan and the effect of maternal and neonatal factors on hematopoietic stem cell content.

BACKGROUND: Cord blood transplant is an accepted treatment for many malignant and non-malignant diseases. We sought to determine the feasibility of collecting cord blood in Jordan and the effect of maternal and fetal factors on the quality of the cord blood units. METHODS: A total of 124 cord blood units were collected, and 75 (60%) cord blood units were included in this analysis. Cord blood volume, total nucleated cell (TNC) count, cell viability and CD34(+) content were measured, and clonogenic assay was performed. RESULTS: The mean volume of the collected units was 68.9 ml (range 40-115) with mean nucleated cell count of 6.5 x 10(8) (range 1-23.0). Our results showed a positive correlation between the volume of cord blood and TNC count (p=0.008), cell viability (p=0.001), CD34(+) content (p=0.034) and the length of the umbilical cord (p=0.011). In addition, our results showed an inverse relation between the Colony Forming Unit-Granulocyte Macrophage (CFU-GM) concentration and the gestation duration (p=0.038). CONCLUSION: We conclude that it is feasible to collect cord blood units in Jordan with excellent TNC and CD34(+) cell content. The volume of cord blood collected was associated with higher TNC count and CD34(+) count. Efforts toward establishing public cord blood banks in our area are warranted.

Investigation of Helicobacter pylori infection in Jordanian patients using six enzyme immunoassays for immunoglobulin G (IGG) and IGA testing.

Helicobacter pylori (H. pylori) is the etiologic agent of a variety of gastrointestinal disorders. The rationale of the current study is to evaluate six enzyme immunoassays for detection of anti-H. pylori immunoglobulin G (IgG) and IgA in Jordanian patients. Biopsy specimens and blood samples were obtained from patients underwent the endoscopy unit at Al-Bashir hospital in Jordan. The serum samples were investigated for the presence of anti-H. pylori IgG and IgA antibodies in patients with positive H. pylori biopsy samples. The results showed that IgG utilizing kits are more sensitive than of IgA kits and the IgA kits are more specific than of that IgG kits. Moreover, the biopsy is seemingly the gold standard for diagnosis of H. pylori is followed by H. pylori culture on brucella agar medium. An imperfect relation between the presence of H. pylori infection and the antibody response was existed that could be explained either because of the unsatisfactory sensitivities and specificities of the commercial kits used or because of weak immunological response in our patients to H. pylori antigens. Collectively, the H. pylori diagnosis that depends on the detection of anti-H. pylori antibodies in the hospital setting and in the screening programs should consider another test for confirmation the initial diagnosis.

Modeling the effects of prenatal exposure to aspirin on the postnatal development of rat brain.

Three growth models were used to examine the effects of prenatal exposure to aspirin on the postnatal development of brain parts. A total of 60 pregnant rats which were divided into three experimental groups and a control group were exposed to aspirin doses of 12.5, 25, 37.5 mg/kg, and distilled water, respectively. The brain parts of 200 rat pups starting from the first week after birth until the fifth week were weighted and the length and width of the cerebrum and cerebellum were measured to determine the parameters of the growth models. The results indicated that the three models successfully predicted the growth of the different brain parts and that aspirin decreased the total brain weight, cerebrum length and width, and decreased the cerebellum length and width at aspirin dose of 37.5 mg/kg. Further analysis is needed to investigate if aspirin effects were carried out through its role in inhibiting prostaglandin production and consequently affecting the activity of the hypothalamus-pituitary axis.

T helper cell polarization and the generation of varied antigen-dependent immune responses: a Markov chain model.

BACKGROUND: Adaptive immune responses are deterministically classified into humoral or cell-mediated depending on the pattern of Th cell polarization into Th1 or Th2. Evidence suggests that the process of Th polarization is stochastic, however, the presence of some deterministic components has not been ruled out. Here, a Markov chain model that accounts for Th-mediated immune responses was developed based on the assumption that Th polarization and consequent transition events are stochastic. RESULTS: Using assumed probability values, model analysis suggests that there is a rapid convergence to produce an immune response once the Th cell is stimulated by an antigen which is amplified as the number of transitions increases. The expected number of visits between Th and itself, B and itself and Tc and itself is about one whereas it is zero, less than one or degrees in the rest of the transition events depending on the interacting states. CONCLUSIONS: Based on model analysis and validation, modeling Th-mediated immune responses as a Markov chain process seems to be plausible. The large degree of flexibility inherent in such a view of adaptive immunity can be helpful in addressing questions pertinent to Th function and behavior.

Modeling acquired immunity as an outcome of the interaction between host-related factors and potential antigen repertoires.

In an attempt to understand why different organisms defend against potential antigens differently, the influence of possible interactions between host-related factors and respective antigen repertoires on the complexity of host defense mechanisms was investigated. A compartmental model coupling these two variables was developed and tested. Data analysis suggests that the more complex the organism, the larger the size of its antigen repertoire. The two variables seem to advance in a parallel fashion suggesting that they could reach a state of equilibrium. Therefore, host-related factors may play a role in determining the size of the antigen repertoire on the one hand; on the other hand, increased antigen repertoire size may dictate the evolution of more complex mechanisms of immunity. Although the interplay between the two variables maintains some common themes in different groups of organisms, it results in clear differences pertinent to immunologic specificity, diversity, memory and self nonself discrimination.

African Jordanian population genetic database on fifteen short tandem repeat genetic loci.

AIM: To establish a genetic database of the African-Jordanian population for forensic and paternity testing purposes. METHOD: Allelic distribution at fifteen short tandem repeat (STR) loci was determined for 95 healthy unrelated African-Jordanians. The 15 autosomal STR loci, included within the GenePrint PowerPlex 16 system, were amplified from the subset of the 95 DNA extracts isolated from the population sample. Electrophoresis for each polymerase chain reaction (PCR) product was carried out using the ABI Prism 310 Genetic Analyzer and the length of the amplified DNA fragments was determined using the Genotype 2.0 and PowerTyper 16 Macro softwares. Calculations of allelic frequencies, forensic efficiency parameters, Hardy-Weinberg departure, and quantitative analysis of the allele frequencies in various populations were determined. RESULTS: DNA extracts were successfully amplified and the genetic database was compiled. All tested loci showed no significant statistical deviation from Hardy-Weinberg expectations. Furthermore, no significant difference was observed between the sample population under investigation and other population genetic databases. CONCLUSION: The loci investigated here proved to be sufficiently polymorphic for forensic purposes, since the forensic efficiency values suggest that they are very discriminating in the African-Jordanian subpopulation.

Isolation and characterization of Candida spp. in Jordanian cancer patients: prevalence, pathogenic determinants, and antifungal sensitivity.

The presence of Candida spp. in the oral cavity was evaluated in 95 cancer patients (57 in-patients and 38 out-patients) and in 65 healthcare workers in Amman, Jordan. Candida carriage occurred in 72.6% of cancer patients and 33.8% of healthcare workers, with Candida albicans being the species most commonly recovered, followed by C. glabrata. In-patients were found to harbor Candida spp. at significantly higher levels than out-patients (P = 0.0044). The number of adhered C. albicans cells and the secretion of extracellular proteinase was significantly higher in the in-patient group than in the out-patient group (P = 0.0016 and 0.00007, respectively); this significant difference was not observed regarding phospholipase secretion. Antifungal sensitivity testing data suggest that isolates were most sensitive to amphotericin B and nystatin, and least sensitive to miconazole and fluconazole, which are commonly used antifungal agents in Jordan.