Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance

Leukemogenesis occurs under hypoxic conditions within the bone marrow (BM). Knockdown of key mediators of cellular responses to hypoxia with shRNA, namely hypoxia-inducible factor-1α (HIF-1α) or HIF-2α, in human acute myeloid leukemia (AML) samples results in their apoptosis and inability to engraft, implicating HIF-1α or HIF-2α as therapeutic targets. However, genetic deletion of Hif-1α has no effect on mouse AML maintenance and may accelerate disease development. Here, we report the impact of conditional genetic deletion of Hif-2α or both Hif-1α and Hif-2α at different stages of leukemogenesis in mice. Deletion of Hif-2α accelerates development of leukemic stem cells (LSCs) and shortens AML latency initiated by Mll-AF9 and its downstream effectors Meis1 and Hoxa9. Notably, the accelerated initiation of AML caused by Hif-2α deletion is further potentiated by Hif-1α codeletion. However, established LSCs lacking Hif-2α or both Hif-1α and Hif-2α propagate AML with the same latency as wild-type LSCs. Furthermore, pharmacological inhibition of the HIF pathway or HIF-2α knockout using the lentiviral CRISPR-Cas9 system in human established leukemic cells with MLL-AF9 translocation have no impact on their functions. We therefore conclude that although Hif-1α and Hif-2α synergize to suppress the development of AML, they are not required for LSC maintenance.Oxygen measurement in the BM indicated that normal and malignant hematopoiesis occur under hypoxic conditions (Spencer et al., 2014). Hif-1 and Hif-2 are key mediators of cellular responses to hypoxia and regulate gene expression to facilitate adaptation to low oxygen tension (Semenza, 2014). Oxygen-regulated α-subunits of Hif-1 and Hif-2, namely Hif-1α and Hif-2α, are paralogs that have common and also distinct functions during responses to hypoxia. Several studies investigated the role of Hif-1α (Wang et al., 2011; Velasco-Hernandez et al., 2014) and Hif-2α (Rouault-Pierre et al., 2013) in acute myeloid leukemia (AML; Gezer et al., 2014; Vyas, 2014). HIF-1α or HIF-2α knockdown in AML patient samples compromised their ability to reconstitute AML upon transplantation into recipient mice (Wang et al., 2011; Rouault-Pierre et al., 2013). Although the subtype and molecular classification of AML samples used in these studies were not specified, the authors implied that HIF-1 and HIF-2 are independently required for the maintenance of AML leukemic stem cells (LSCs), suggesting that HIF-1 and HIF-2 are potential therapeutic targets for AML (Wang et al., 2011; Rouault-Pierre et al., 2013).Considering the caveats of shRNA-mediated gene knockdown approaches, a recent study used a conditional Hif-1α knockout and reported that, surprisingly, conditional Hif-1α deletion does not compromise the development and maintenance of mouse LSCs generated by the MLL-ENL fusion, its downstream effectors Meis1 and Hoxa9, and Aml1-Eto9a (Velasco-Hernandez et al., 2014). In fact, loss of Hif-1α accelerated the development of Meis1/Hoxa9-induced AML or enhanced propagation of disease induced by Aml1-Eto9a (Velasco-Hernandez et al., 2014). This study concluded that Hif-1α can suppress LSC development or propagation and is dispensable for AML LSC maintenance, sparking a debate over the therapeutic benefit of targeting HIF-1 function (Vyas, 2014).To date, the impact of conditional deletion of Hif-2α or loss of both Hif-1α and Hif-2α on leukemic transformation has not been examined. Therefore, in this study, we set out to investigate the requirement for Hif-2α or both Hif-1α and Hif-2α in the development and maintenance of AML LSCs.     

Histone deacetylase inhibitor-mediated sensitization to TRAIL-induced apoptosis in childhood malignancies is not associated with upregulation of TRAIL receptor expression, but with potentiated caspase-8 activation

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has great potential for the treatment of cancer because it targets tumor cells while sparing normal cells. Several cancers, however, fail to respond to TRAIL's antineoplastic effects. These resistant tumors require cotreatment with sensitizing agents in order for TRAIL to exert anticancer activity. Histone deacetylase inhibitors (HDACi) have been recognized as potent TRAIL sensitizers. In searching for the determinants of TRAIL responsiveness, HDACi-mediated TRAIL sensitization has been predominantly attributed to TRAIL receptor upregulation. This explanation, however, has been challenged by a few studies. The aim of the present study was to explore the relevance of TRAIL receptor expression for HDACi-mediated TRAIL sensitization in childhood tumors, i.e., in medulloblastoma, Ewing's sarcoma and osteosarcoma. In previous studies, we had shown that TRAIL and HDACi were synergistic in inducing apoptosis in medulloblastoma and Ewing's sarcoma. In the present study, we demonstrate that HDACi cooperated with TRAIL in eliciting cell death in osteosarcoma. However, HDACi treatment did not alter or even reduced cell surface expression of TRAIL receptors in the three childhood tumors. In gaining insight into the apoptotic pathway involved in TRAIL sensitization, HDACi were found to potentiate TRAIL-induced caspase-8 activation. Taken together, our findings suggest that HDACi-mediated TRAIL sensitization is not the result of TRAIL receptor upregulation, but the result of a receptor-proximal event in childhood tumor cells.     

Genital malodor in women: a modern reappraisal

Genital malodor is a common distressing complaint that brings a woman to her physician's office. Vaginal infections, primarily bacterial vaginosis and trichomoniasis, still remain the commonest causes and are relatively easy to diagnose and treat. However, in approximately one third of women who present with malodor, no cause is identified. Although data on the management of vaginal discharge are extensive, the management of genital odor beyond common vaginal infections remains poorly studied. This presents a frustrating situation for both the patient and her physician. Often, patients resort to home remedies and over-the-counter preparations, which, while providing short-term relief for some women, almost never address the cause and, in some cases, can exacerbate symptoms. In this review, we have attempted to consolidate the known and documented causes of genital malodor including the nonvaginal causes and provide case studies that will help clinicians understand the possible settings for the various causes. We also provide an algorithm for the management of this symptom beyond vaginal infections.     

To Assess the Effect of Trauma on the Temporomandibular Joint in Postoperative Cases of Zygomaticomaxillary Complex Fractures

Aim and ObjectivesThe study aims to assess the incidence and features of temporomandibular joint(TMJ) dysfunction in post-surgical treatment of unilateral zygomaticomaxillary complex(ZMC) fractures.The objectives are:

1. To assess severity of TMJ dysfunction in postoperative cases of ZMC fractures.
2. To create awareness of the same among clinicians.

MethodsPatients presenting with zygomaticomaxillary complex fractures were evaluated prospectively. Evaluation of TMJ dysfunction was done by different parameters via questionnaire, clinical and radiographic examination preoperatively and a follow-up period of 1 week, 3 months and 6 months. The parameters were, clicking of joint, pain on opening /closing, pain on biting, deviation of mandible, pain in the preauricular region, ringing sound and mouth opening. Statistical analysis was done by the Friedman test and Post Hoc analysis.ResultsOn presentation, 69.1% patients diagnosed with ZMC fractures presented with symptoms related to TMJ dysfunction. Post-surgery 1 week majority findings persisted, with 21 patients complained of pain on opening or closing and 2 patients with a persistent opening click. These symptoms, however, decreased over the 3 month and 6 month follow up period. 5 patients presented with decreased mouth opening which was attributed to lack of adequate physiotherapy.ConclusionPatients presented with mild symptoms of TMJ dysfunction until 6 months post-surgery, however these symptoms weren’t significant as the pain score assessed was found to decrease in the following post-operative periods. And the symptoms present were’nt exclusive to conclude a TMJ dysfunction. Early treatment and a close follow up are key to prevent progression of symptoms.     

A bovine parainfluenza virus type 3 vaccine is safe and immunogenic in early infancy

BACKGROUND: A phase 2 trial was conducted to assess in young infants the safety, tolerability, infectivity, and immunogenicity of multiple doses of an intranasal vaccine using bovine parainfluenza virus type 3 (bPIV3). METHODS: One hundred ninety-two healthy 2-month-old infants were randomized 1 : 1 : 1 to receive 1x10(5) median tissue culture infective dose (TCID(50)) bPIV3 vaccine, 1x10(6) TCID(50) bPIV3 vaccine, or placebo at 2, 4, 6, and 12-15 months of age. Safety information was collected by use of diary sheets and telephone interviews. Nasal wash and serum specimens were collected for assessment of infectivity and immunogenicity. RESULTS: The safety profiles of both dosages of bPIV3 were similar to that of placebo, with the exception of fever with temperature of >/=38.1 degrees C after dose 2 only, occurring in 34% of the 1x10(5) TCID(50) group, 35% of the 1x10(6) TCID(50) group, and 12% of the placebo group (P<.01). No vaccine-related serious adverse events were reported. The cumulative vaccine infectivity (isolation of bPIV3 and/or bPIV3 seroconversion) after dose 3 was similar in the 2 vaccine groups (87% in the 1x10(5) TCID(50) group and 77% in the 1x10(6) TCID(50) group) (P=.46). Seroconversion rates after dose 3, assessed by means of hemagglutination inhibition assay, after adjustment for decrease in maternal antibody titers, were 67% in the 1x10(5) TCID(50) group, 57% in the 1x10(6) TCID(50) group, and 12% in the placebo group (P<.01). Isolation of bPIV3 was common after dose 1, dose 2, or dose 3, but only 1 of 51 participants in the vaccine groups had bPIV3 isolated after dose 4. CONCLUSIONS: Multiple doses of bPIV3 vaccine were well tolerated and immunogenic in young infants.     

SPME-GC determination of potential volatile organic leachables in aqueous-based pharmaceutical formulations packaged in overwrapped LDPE vials

A direct liquid immersion solid-phase microextraction-gas chromatographic (SPME-GC) method was developed and validated for the determination of 11 potential volatile organic compounds that may leach from preprinted foil laminate overwrap into aqueous pharmaceutical formulations filled in low-density polyethylene (LDPE) vials. The target compounds namely, ethanol, acetone, isopropyl alcohol, ethyl acetate, 2-butanone, n-heptane, isopropyl acetate, n-propyl acetate, toluene, diacetone alcohol and 1-propoxy-2-propanol, were suitably extracted from aqueous sample solutions by SPME using a 100-microm PDMS fiber, desorbed inside the GC inlet port, and analyzed using a J&W Scientific DB-1701 (86% polydimethylsiloxane/14% cyanopropylphenyl, 30 m x 0.53 mm i.d., 1.5-microm film thickness) capillary column with FID detection. The variables affecting the SPME absorption and desorption conditions were optimized and discussed. The average recoveries for all analytes varied from about 97.9 to 116.7% with the exception of n-heptane and toluene where the mean recoveries ranged from about 73.6 to 100.0% presumably due to their poor solubility in the aqueous sample matrix. The standard curves for all compounds were linear over the concentration range investigated with coefficient of correlations, r(2)> or =0.98. The detection and quantitation limits ranged from approximately 0.6 ng/ml to 1.7 microg/ml and 5 ng/ml to 4.2 microg/ml, respectively, and the intra- and inter-day precision was considered adequate (R.S.D.< or =16%) for low-level determination of the target analytes in the sample matrix. The method was successfully applied for determination of the target compounds from preprinted foil laminate overwrap in selected aqueous-based pharmaceutical formulations.     

Effectiveness of myofascial release in the management of lateral epicondylitis in computer professionals

Ajimsha MS, Chithra S, Thulasyammal RP. Effectiveness of myofascial release in the management of lateral epicondylitis in computer professionals.ObjectiveTo investigate whether myofascial release (MFR) reduces the pain and functional disability of lateral epicondylitis (LE) in comparison with a control group receiving sham ultrasound therapy in computer professionals.DesignRandomized, controlled, single blinded trial.SettingNonprofit research foundation clinic in Kerala, India.ParticipantsComputer professionals (N=68) with LE.InterventionsMFR group or control group. The techniques were administered by certified MFR practitioners and consisted of 12 sessions per client over 4 weeks.Main Outcome MeasureThe Patient-Rated Tennis Elbow Evaluation (PRTEE) scale was used to assess pain severity and functional disability. The primary outcome measure was the difference in PRTEE scale scores between week 1 (pretest score), week 4 (posttest score), and follow-up at week 12 after randomization.ResultsThe simple main effects analysis showed that the MFR group performed better than the control group in weeks 4 and 12 (P<.005). Patients in the MFR and control groups reported a 78.7% and 6.8% reduction, respectively, in their pain and functional disability in week 4 compared with that in week 1, which persisted as 63.1% in the follow-up at week 12 in the MFR group.ConclusionsThis study provides evidence that MFR is more effective than a control intervention for LE in computer professionals.     

An Erupted Silent Tumour

Odontomas are the most common odontogenic tumors of the oral cavity which are nonaggressive, hamartomatous in nature consisting of enamel, dentin and cementum. They are called as composite because they contain more than one type of tissue. They are generally asymptomatic, hence recognised on routine radiologic examination. The compound odontoma is composed of multiple small tooth like structures, whereas the complex odontoma consists of a conglomerate mass of enamel and dentine, which bears no anatomical similarity to the tooth. The eruption and infection of odontoma are uncommon, only few cases of erupted complex odontoma are reported in the literature. We report a case of silent erupting complex odontoma.     

GLUT9 is differentially expressed and targeted in the preimplantation embryo

During preimplantation development in the mouse, it is crucial that glucose metabolism not be compromised. Any decrease in glucose uptake at this stage in development can compromise the developing embryo. We have cloned another member of the glucose transporter family, GLUT9, which is expressed embryonically. Three different isoforms were identified. We have shown that two of the mouse GLUT9 isoforms transport glucose at a rate significantly greater than controls. Expression analysis of the preimplantation blastocyst identifies only the presence of the shorter GLUT9 isoform, RT-PCR and Western immunoblot confirmed this finding. A differential pattern of expression was seen with GLUT9 present at the plasma membrane in one- and two-cell zygotes and in an intracellular compartment in trophectoderm cells at a blastocyst stage. Although blocking GLUT9 expression during preimplantation development had no effect on glucose transport or apoptosis, transfer of these embryos into pseudopregnant mice resulted in increased pregnancy loss, suggesting that GLUT9 is critical for early preimplantation development.