[Inhibition of Histone Deacetylases Reverses Epithelial-Mesenchymal Transition in Triple-Negative Breast Cancer Cells through a Slug Mediated Mechanism].

High metastatic ability and poor clinical outcome are the most known clinical features of the triple-negative breast tumors. Given that the tumor cells undergoing epithelial-mesenchymal transition (EMT) often gain malignant and invasive features, we have investigated the possibility of EMT reversal in triple-negative breast cancer cells by targeting the epigenetic-modifying enzymatic complexes named histone deacetylases (HDACs) and examined the possible mechanism underlying the HDACs-based inversion in model MDA-MB-231 cells. Cells were treated with a maximal tolerable 200 nM concentrations of classical HDACs inhibitor Trichostatin A (TSA) for 48 h and afterwards the invasiveness and immigration of the cells were evaluated in TransWell Invasion Scratch Wound Healing assays. Then, in treated and control cells, quantitative real time-PCR reacions were performed for assessing the gene expression of EMT biomarkers E-cadherin, Vimentin and transcriptional factor Slug. After TSA treatment, the invasion and migration properties MDA-MB-231 cells significantly decreased, gene expression of E-cadherin was significantly up-regulated, while the levels of Slug and Vimentin encoding mRNAs were suppressed. We conclude that inhibition of HDACs in triple-negative breast cancer cells may lead to inversion of EMT and the decrease of invasiveness by down-regulating the gene expression of Slug. Since EMT is known as a pre-metastatic process, triple-negative breast tumors, the EMT reversal effects of HDACs inhibition may reduce tumor cell metastasis.

TERT promoter mutations in gliomas, genetic associations and clinico-pathological correlations.

BACKGROUND: The role of telomerase reverse transcriptase (TERT) in gliomagenesis has been recently further strengthened by the frequent occurrence of TERT promoter mutations (TERTp-mut) in gliomas and evidence that the TERT SNP genetic rs2736100 influences glioma risk. TERTp-mut creates a binding site for Ets/TCF transcription factors, whereas the common rs2853669 polymorphism disrupts another Ets/TCF site on TERT promoter. METHODS: We sequenced for TERTp-mut in 807 glioma DNAs and in 235 blood DNAs and analysed TERT expression by RT-PCR in 151 samples. TERTp-mut status and TERTp polymorphism rs2853669 were correlated with histology, genomic profile, TERT mRNA expression, clinical outcome and rs2736100 genotype. RESULTS: TERTp-mut identified in 60.8% of gliomas (491 out of 807) was globally associated with poorer outcome (Hazard ratio (HR)=1.50). We defined, based on TERTp-mut and IDH mutation status, four prognostic groups: (1) TERTp-mut and IDH-mut associated with 1p19q codeletion, overall survival (OS)>17 years; (2) TERTp-wt and IDH-mut, associated with TP53 mutation, OS=97.5 months; (3) TERTp-wt and IDH-wt, with no specific association, OS=31.6 months; (4) TERTp-mut and IDH-wt, associated with EGFR amplification, OS=15.4 months. TERTp-mut was associated with higher TERT mRNA expression, whereas the rs2853669 variant was associated with lower TERT mRNA expression. The mutation of CIC (a repressor of ETV1-5 belonging to the Ets/TCF family) was also associated with TERT mRNA upregulation. CONCLUSIONS: In addition to IDH mutation status, defining the TERTp-mut status of glial tumours should afford enhanced prognostic stratification of patients with glioma. We also show that TERTp-mut, rs2853669 variant and CIC mutation influence Tert expression. This effect could be mediated by Ets/TCF transcription factors.

Probiotic inhibits the cytopathic effect induced by Escherichia coli O157:H7 in Vero cell line model.

AIMS: The effectiveness of four strains of Bifidobacteria against enterohemorrhagic Escherichia coli O157:H7 infection was studied using a Vero cell model. METHODS AND RESULTS: E. coli O157 was inoculated on the Vero cell line before and after treatment with probiotic. The cytopathic effect (CPE) was evaluated during 24 h of incubation. The results indicated that Shiga toxin activity was inhibited by the probiotic. To prevent a Stx2 CPE, the probiotic needs one log more than the Stx1. CONCLUSION: The Vero cell assay, in particular, is a good model to evaluate the effect of Bifidobacteria inhibiting bacterial attachment because of soluble substances and the competitive aspect and could be used in a variety of foods like milk and yoghurt to protect pathogen bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Probiotics could control pathogenic bacteria and Vero cell introduce as a model for evaluation of probiotics against pathogen bacteria.

Medullary thyroid cancer is associated with high serum vitamin D level and polymorphism of vitamin D receptors.

BACKGROUND: Thyroid cancer is the most common endocrine malignancy. Studies have observed an anti-cancer effect for vitamin D and found that polymorphisms of vitamin D receptors can influence the prevalence of various cancers. The present study investigated the serum level of vitamin D and FokI, BsmI and Tru9I polymorphisms of vitamin D receptors. METHODS: Forty patients with medullary thyroid cancer and 40 healthy controls were investigated. The genomic DNA of the subjects was extracted using saturated salt/proteinase K and investigated by PCR sequencing. Serum levels of vitamin D were evaluated by ELISA. The results were analyzed in SPSS and GraphPad Prism 5 software. RESULTS: The genotypic and allelic frequencies of FokI and BsmI polymorphisms showed no significant differences between test and control groups. For Tru9I polymorphism, Tt genotype and t allelic frequency in the test group were significantly different from those of the control group. Also, we found Tt genotype and t allelic frequency to be significantly associated with medullary thyroid cancer (MTC) type and the agressiveness of the disease. The average serum vitamin D level was 23.32 ng/mL and 18.95 ng/mL for patients and controls, respectively, and the difference between the two groups was statistically significant. Moreover, we found high serum vitamin D level to be associated with t allelic frequency. CONCLUSIONS: Unexpectedly, the mean serum vitamin D level of the test group was significantly higher than that of the control group. Tru9I polymorphism was found to be significantly correlated with the prevalence of medullary thyroid carcinoma.

Experimental Neospora caninum infection in domestic bird's embryonated eggs.

To date, there are no reports regarding comparison between different bird species in Neospora. caninum infection. In the present study 70 embryonated eggs from quail, partridge, broiler and egg laying chickens were divided into 7 groups equally. Six groups in each species were inoculated with different dilutions (10, 10(2), 10(3), 10(4), 10(5), and 10(6)) of tachyzoites/embryonated egg in the chorioallantoic membrane and the seventh group was considered as control. The mortality rates and clinical signs were studied. All the egg laying chickens and some of the broiler chickens and quails showed neurologic signs like. The results revealed that the mortality rate was dose dependent in broiler chicken embryonated eggs. But mortality rate was dose independent in egg laying chickens and quail. Partridge revealed 100 % mortality rate in all doses. The LD50 in broiler chicken embryonated was 10(2.3). In conclusion, LD50 in the broiler chickens is the lowest between different animal models which shows that the broiler chicken embryonated egg is the best animal model for experimental inducing of neosporosis. Partridge is the most susceptible bird to N. caninum infection. These results reinforce that there is genetic susceptibility to N. caninum in chickens like mice and provide new insights to reach an inexpensive and available animal model for N. caninum infection.

First isolation and establishment of in vitro culture of Theileria lestoquardi from a naturally infected cow.

Theileria infected cell line was isolated from the prescapular lymph node of an adult crossbred cow. Molecular study confirmed this cell line of bovine lymphocyte has been transformed by the Theileria lestoquardi. This strain of T. lestoquardi designated Ka-6 and sheep were inoculated with this strain didn't show any clinical signs of theileriosis which shows the significance of this cell line to develop a tissue-culture vaccine against malignant ovine theileriosis. Contrary to accepted belief that the T. lestoquardi not capable of causing disease in cattle, the present study describes the first isolation and establishment of in vitro culture of T. lestoquardi-infected cell line from a naturally infected cow with typical singes of acute theileriosis.

A randomized trial comparing standard outcomes in two treatment models for substance users with tuberculosis.

SETTING: Chicago Department of Public Health (CDPH), TB Control Program. OBJECTIVES: To compare anti-tuberculosis treatment outcomes using two different types of directly observed therapy (DOT) outreach workers. METHODS: Substance users diagnosed with TB from October 1996 to July 2000 were randomized to DOT administered by either 1) CDPH personnel (standard arm) or 2) previous substance-using human immunodeficiency virus/acquired immune-deficiency syndrome outreach workers (enhanced arm). Treatment completion was physician-determined, and adherence was estimated based on risk of missed DOT appointments. RESULTS: Of 94 patients, 46 were randomized to the standard and 48 to the enhanced arm. The standard arm had a significantly higher risk of non-completion of treatment (39% vs. 15%, RR 2.7, 95%CI 1.2-5.8), and a significantly higher risk of missing DOT appointments (RR 2.6, 95%CI 1.4-4.8). For both outcomes, housing instability was a significant predictor in multivariate analyses. CONCLUSIONS: TB treatment completion and adherence among substance users was improved by the enhanced intervention; the familiarity of enhanced-arm DOT workers with the patients' social norms due to their own previous substance use may have made them more effective. Successful DOT in hard-to-reach populations may require strategies that directly address the population's circumstances and utilize DOT workers who are intimately familiar with patients' life situations.

The onset of drug injection among sex partners of injection drug users.

HIV prevention interventions targeting noninjecting sex partners of drug injectors usually focus on sexual behaviors. This strategy may underestimate the likelihood that sex partners will begin injecting drug and thereby greatly increase their exposure to HIV. This 4-year prospective study assesses the incidence of drug injection among 62 street-recruited, heterosexual, HIV negative, baseline noninjection sex partners, 97% of whom were reinterviewed at least once. Sixteen (26%) of the sex partners reported injecting after baseline. Thirty-nine percent of those with no history of illicit drug use beyond marijuana began injecting, compared with 19% of those who had used drugs but had never injected and 11% of those with injection histories. There were two HIV seroconversions, both of which took place soon after the onset of injection and appear attributable to parenteral transmission. These results suggest that interventions targeting sex partners should include strategies for preventing injection and risky injection practices.

Effects of HIV/AIDS education and training on patient care and provider practices: a cross-cutting evaluation.

Initial and continuing training in HIV/AIDS service provision is a critical way to enable the nation's health providers to use state-of-the-art developments and perspectives. Typically, the efficacy of HIV/AIDS training programs is evaluated using assessments administered to trainees immediately following the training. This study reports cross-cutting findings from telephone interviews conducted with 218 trainees an average of 8 months after training. Long-term training effects are examined in three domains: (a) general perspectives on HIV/AIDS; (b) health care provider service provision; and (c) changes in procedures and operations at the health care system level. The findings show the different ways that the training experience had long-term positive and observable effects in these three domains. In some cases, background characteristics and job positions predicted the specific type of reported training effects. The pattern of results suggests ways in which training methods can be targeted to specific audiences.

Systems change resulting from HIV/AIDS education and training. A cross-cutting evaluation of nine innovative projects.

An evaluation of nine diverse HIV/AIDS training programs assessed the degree to which the programs produced changes in the ways that health care systems deliver HIV/AIDS care. Participants were interviewed an average of 8 months following completion of training and asked for specific examples of a resulting change in their health care system. More than half of the trainees gave at least one example of a systems change. The examples included the way patient referrals are made, the manner in which agency collaborations are organized, and the way care is delivered.

A model of homeless migration: homeless men in Skid Row, Los Angeles.

"In this paper a conceptual model of homeless migration is developed that links migration behavior with the coping status of homeless individuals. The model is evaluated by using data drawn from a recent random probability sample of men surveyed in Skid Row, Los Angeles, [California]. Results indicate that homeless migrants tended to be young, never married, white, mentally disabled, and either newly or cyclically homeless individuals. Long-term residents, in contrast, were apt to be older, physically disabled or suffering from a health-related problem, and had been homeless for some time. The dominant reason given for moving was to find a job...." Coping strategies and migration behaviors are also described.

Suspension culture of Neospora caninum by Theileria annulata-infected cell line.

There are some limiting aspects of scaling up the Neospora caninum tachyzoites in continuous cell lines, particularly as related to the absence of surface attachment. In this study, suspension cell culture of Theileria annulata-infected lymphoblastoid (TIL) was used as a host cell for the continous production of N. caninum tachyzoites. The numbers of free tachyzoites in the medium supernatant were showed regularly increased up to the day 6 post-cultivation. Transmission electron microscopy demonstrated that N. caninum tachyzoites invaded the TIL cells and multiplied intracellularly. This showed that the tachyzoites were successfully proliferated in TIL cells and were released in complete Dulbecco's modified Eagle's medium. This is a successful report of in vitro cultivation of N. caninum tachyzoites achieved by using suspension host cell culture.

Evaluation of Cross Immunity and Histopathological Findings in Experimentally Infected BALB/c Mice with Neospora caninum and Besnoitia caprae.

BACKGROUND: Caprine besnoitiosis is an economically important disease of goats. Neospora caninum, another coccidian parasite of worldwide distribution, infects several animal species and is a major cause of abortion in cattle. Combined infections of N. caninum and Besnoitia caprae can occur in geographical areas endemic for both species of parasite in goats. This experiment was conducted to investigate the possible cross-immunity between these two infections in experimentally infected BALB/c mice. METHODS: Forty BALB/c mice were divided into four equal groups. The mice of Groups 1 and 4 were inoculated with 1×10(6) live virulent tachyzoites of N. caninum (NC-1), while animals of Groups 2 and 3 were inoculated with sterile tissue culture medium. Each mouse in Groups 1 and 2 was challenged 28 days later with 1×10(6) live virulent bradyzoites of B. Caprae (BC-1). RESULTS: Following the challenge, the mice in Groups 1 and 2 showed 100% morbidity and 100% mortality within 9 days post infection, while all the animals of Groups 3 and 4 remained alive. The dead animals were necropsied. The survivors (mice in Group 3 and 4) were euthanized 9 days after inoculation and the gross and histopathological lesions in different organs were investigated. CONCLUSION: Immunization and challenge experiments with lethal dose of B. caprae in the highly susceptible BALB/c mice showed no cross-protection between N. caninum and B. caprae.

Megakaryocytic differentiation of CD133+ hematopoietic stem cells by down-regulation of microRNA-10a.

MicroRNAs (miRNA), as a class of non-coding RNAs, play a crucial role in normal hematopoiesis. Recent studies on miRNA profiling during hematopoiesis showed miR-10a down-regulation in megakaryocytic differentiation. Here we investigated whether decrease in miR-10a can differentiate umbilical cord blood CD133+ cells to megakaryocytic series. We ectopically induced miR-10a down-regulation by locked nucleic acid anti-miR-10a transfection of CD133+ cells. The megakaryocytic differentiation was evaluated by CD42/CD61 and CD41 surface expression and colonogenic capacity in Megacult media. In addition, real-time polymerase chain reaction was done for detection of miR-10a and its target HoxA1 mRNA. HOXA1 protein expression was detected by flow cytometry as mean fluorescent intensity that shows the antibody reaction proteins. Our findings showed megakaryocytic differentiation of about 28% of umbilical cord blood CD133+ stem cells with bright expression of CD42/CD61 and CD41 in parallel with increase in HoxA1 mRNA and protein level. Colony forming of CD61+ cells in Megacult demonstrated the colonogenic capacity of differentiated progenitor cells. In conclusion, MiR-10a has a role in megakaryocyte differentiation of stem cells via HOXA1 transcription factor targeting.

Seroprevalence of Neospora spp. in Horses in North East of Iran.

BACKGROUND: Neospora caninum, an obligate intracellular protozoan parasite, is recognized as a major cause of abortion in cattle, while limited information is presently available on the seroprevalence of Neospora antibodies in horses' worldwide. The aim of the present study was to determine serologic prevalence of Neospora infection in horses in Iran. METHODS: Sera from 150 horses from Mashhad suburb in Razavi Khorasan Province, northeast Iran were examined for antibodies to Neospora spp. using Neospora modified direct agglutination test (N-MAT). RESULTS: Antibodies to this parasite were detected in 45 (30%) of the examined serum samples. Thirty four percent of the samples had titer of 1:40 while then reduced to 30% when 1:80 serum dilution was applied as significant cut off titer. CONCLUSION: This study is the first investigation carried out on the Neospora in horses in Iran and indicates that horses in Iran are exposed to this parasite.

Seroprevalence of Neospora spp. in horses in South of Iran.

Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. However, limited information is presently available on the seroprevalence of Neospora antibodies in horses worldwide. The aim of the present study is to determine serological prevalence of Neospora infection in horses in Iran. Blood samples were obtained from 200 horses and tested for serum antibodies against Neospora spp. by the Neospora modified direct agglutination test (N-MAT). Antibodies were found in 64 (32%) horses being tested with titers of 1:80. This is the first serological survey for Neospora antibodies performed on horses in Iran.

Bypassing the maturation arrest in myeloid cell line U937 by over-expression of microRNA-424.

Micro RNAs are a class of small non-coding RNAs which has been recently shown to play a crucial role in major cellular processes such as development and differentiation through post-transcriptional regulation. The role of these epigenetic elements has also been demonstrated in hematopoietic lineage differentiation and there is a large body of evidence that miR-424 is responsible for monocyte differentiation. Our goal was to examine the effect of miR-424 over-expression on defeating the maturation blockage in monoblastic cell line U937. The permanent over-expression of miR-424 was established using a retroviral vector construct containing the precursor of miR-424 sequence. Induction of differentiation process was monitored by assaying changes in cell morphology, and expression of cell surface markers using light microscopy, quantitative RT-PCR, and flow cytometry for monocyte markers such as CD11b and CD14. The cells showed monocytic characteristics 14 days after transduction, and CD11b and CD14 expression were significantly increased, confirmed by flow cytometry QRT-PCR and RT-PCR results. In conclusion, miR-424 over-expression is an effective factor in maturation of the monoblastic U937 cells and it has the ability of directing them into cells, expressing monocyte/macrophage characteristics.

Seroprevalence of Neospora spp. in horses in South of Iran

Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. However, limited information is presently available on the seroprevalence of Neospora antibodies in horses worldwide. The aim of the present study is to determine serological prevalence of Neospora infection in horses in Iran. Blood samples were obtained from 200 horses and tested for serum antibodies against Neospora spp. by the Neospora modified direct agglutination test (N-MAT). Antibodies were found in 64 (32%) horses being tested with titers of 1:80. This is the first serological survey for Neospora antibodies performed on horses in Iran.

Stories of AIDS outreach and case management: context and activities.

Research has shown that injection drug users (IDUs) are now at greater risk for contracting HIV infection. Studies also show that seroincidence has begun to decrease among IDUs in a number of cities due to risk-reduction interventions. One important intervention is the use of indigenous outreach workers, shown to be an effective method in reducing HIV risk behavior and promoting preventive actions among IDUs in various settings. This study explores continuities and changes in the activities of outreach workers and in their changing role as case managers in the long-standing Community Outreach Intervention Project in Chicago. It examines their efforts to change risk behaviors and improve the health and living conditions of IDUs. This research is based on outreach and case-worker perspectives and related background data gathered from 10 outreach workers and the four ethnographers that supervise the project. Outreach and case workers describe the diverse populations and contexts in which they operate and the growing complexity and depth of the issues they face, especially in working with HIV and AIDS-afflicted clients. These descriptions demonstrate the important role "indigenous" outreach and case workers play in engaging out-of-treatment IDUs, supporting meaningful changes in their lives, and responding to their particular and emerging needs.

The maternal and child health sites' practices regarding HIV education, counseling, and testing of women of reproductive age in Chicago: barriers to universal implementation.

OBJECTIVES: Women of reproductive age are increasingly at risk for human immunodeficiency virus (HIV) infection. Recent advances in reducing perinatal transmission have resulted in official guidelines on universal HIV education, counseling, and voluntary testing of women of reproductive age, especially pregnant women. This study assesses to what extent the maternal child health (MCH) sites are implementing these guidelines with their female patient population (including pregnant women) and examines the barriers that prevent them from implementing these guidelines. METHOD: The study uses survey data from 92 hospitals and community health centers offering MCH services in Chicago regarding their providers' practices on HIV education, counseling, and testing, implementation of zidovudine (ZDV) therapy to reduce perinatal transmission, and the barriers to implementing these services. In addition, 20 taped in-depth interviews were conducted with experts to examine the barriers to universal implementation. RESULTS: Almost half (45% of perinatal care and 50% of family planning providers) of the institutions are not consistently offering HIV testing. One-third of those institutions that offer testing are not offering pretest counseling. Thirty-nine percent of the perinatal care providers in these institutions are not providing posttest counseling to HIV-negative women. Over one-third (35%) of these institutions reported that they are not set up to implement ZDV therapy during labor and delivery. Almost half (49%) had no protocols for ZDV therapy in place. Barriers to implementation included lack of provider training, limited staff time, physician resistance, unavailability or avoidance to seek perinatal care by high-risk women, cost, absence of a statewide and hospital-specific plan, lack of reproductive choice focus in posttest counseling, lack of provider knowledge about the administration of ZDV or its availability during labor, and lack of consumer education on perinatal risk reduction. CONCLUSIONS: MCH sites and their providers need assistance to overcome many barriers they face to implement universal HIV education, counseling, and testing of women of reproductive age.