5,5'-Dibromo-bis(3'-indolyl)methane induces Kruppel-like factor 4 and p21 in colon cancer cells

Bis(3'-indolyl)methane (DIM) is a metabolite of the phytochemical indole-3-carbinol, and both compounds exhibit a broad spectrum of anticancer activities. We have developed a series of synthetic symmetrical ring-substituted DIM analogues, including 5,5'-dibromoDIM, which are more potent than DIM as inhibitors of cancer cell and tumor growth. In colon cancer cells, 5,5'-dibromoDIM decreased cell proliferation and inhibited G(0)-G(1)- to S-phase progression, and this was accompanied by induction of the cyclin-dependent kinase inhibitor p21 in HT-29 and RKO colon cancer cells. Mechanistic studies showed that induction of p21 in both RKO (p53 wild-type) and HT-29 (p53 mutant) cells by 5,5'-dibromoDIM was Krüppel-like factor 4 (KLF4) dependent, and induction of p53 in RKO cells was also KLF4 dependent. Analysis of the p21 promoter in p53-dependent RKO cells showed that 5,5'-dibromoDIM activated p21 gene expression through the proximal GC-rich sites 1 and 2, and chromatin immunoprecipitation assays showed that KLF4 and p53 bound to this region of the promoter, whereas in HT-29 cells unidentified upstream cis-elements were required for induction of p21. 5,5'-DibromoDIM (30 mg/kg/d) also inhibited tumor growth and induced p21 in athymic nude mice bearing RKO cells as xenografts, showing that ring-substituted DIM such as 5,5'-dibromoDIM represent a novel class of mechanism-based drugs for clinical treatment of colon cancer.     

Anthocyanin fraction from potato extracts is cytotoxic to prostate cancer cells through activation of caspase-dependent and caspase-independent pathways

Polyphenols from fruits and vegetables exhibit anticancer properties both in vitro and in vivo and specialty potatoes are an excellent source of dietary polyphenols, including phenolic acids and anthocyanins. This study investigated the effects of specialty potato phenolics and their fractions on LNCaP (androgen dependent) and PC-3 (androgen independent) prostate cancer cells. Phenolic extracts from four specialty potato cultivars CO112F2-2, PATX99P32-2, ATTX98462-3 and ATTX98491-3 and organic acid, phenolic acid and anthocyanin fractions (AF) were used in this study. CO112F2-2 cultivar extracts and their AF at 5 mug chlorogenic acid eq/ml were more active and inhibited cell proliferation and increased the cyclin-dependent kinase inhibitor p27 levels in both LNCaP and PC-3 cells. Potato extract and AF induced apoptosis in both the cells and, however, the effects were cell context dependent. Cell death pathways induced by potato extract and AF were associated with mitogen-activated protein kinase and c-jun N-terminal kinase activation and these kinases activated caspase-independent apoptosis through nuclear translocation of endonuclease G (Endo G) and apoptosis-inducing factor in both cell lines. Induction of caspase-dependent apoptosis was also kinase dependent but was observed only in LNCaP cells. Kinase inhibitors reversed this nuclear translocation of endonuclease G and apoptosis-inducing factor. This is the first report showing that the cytotoxic activities of potato extract/AF in cancer cells were due to activation of caspase-independent apoptosis. Current studies are focused on identifying individual components of the AF responsible for the induction of cell death pathways in prostate and other cancer cell lines and developing potato cultivars that overexpress these active compounds.     

A Progressive Mouse Model of Parkinson’s Disease: The Thy1-aSyn (“Line 61”) Mice

Identification of mutations that cause rare familial forms of Parkinson’s disease (PD) and subsequent studies of genetic risk factors for sporadic PD have led to an improved understanding of the pathological mechanisms that may cause nonfamilial PD. In particular, genetic and pathological studies strongly suggest that alpha-synuclein, albeit very rarely mutated in PD patients, plays a critical role in the vast majority of individuals with the sporadic form of the disease. We have extensively characterized a mouse model over-expressing full-length, human, wild-type alpha-synuclein under the Thy-1 promoter. We have also shown that this model reproduces many features of sporadic PD, including progressive changes in dopamine release and striatal content, alpha-synuclein pathology, deficits in motor and nonmotor functions that are affected in pre-manifest and manifest phases of PD, inflammation, and biochemical and molecular changes similar to those observed in PD. Preclinical studies have already demonstrated improvement with promising new drugs in this model, which provides an opportunity to test novel neuroprotective strategies during different phases of the disorder using endpoint measures with high power to detect drug effects.     

Backscattered Dose Perturbation Effects at Metallic Interfaces Irradiated by High-Energy X- and Gamma-Ray Therapeutic Beams

PURPOSE: To analyze backscattered dose enhancements near different metallic interfaces for cobalt-60 ((60)Co) gamma rays and 6- and 18-MV photon beams. MATERIAL AND METHODS: Measurements were carried out with a PTW thin-window, parallel-plate ionization chamber and an RDM-1F electrometer. Thin sheets of aluminum, mild steel, copper, cadmium and lead were used as inhomogeneities. The chamber was positioned below the inhomogenities with the gantry maintained under the couch. RESULTS: It can be noticed that the backscatter dose factor (BSDF) reaches the saturation value within few millimeters of all inhomogeneities and the thickness at which the saturation value is reached depends on the atomic number of the inhomogeneity. The amount of backscattered radiation was noticed to be greater with lesser-energy photons ((60)Co) compared to the high erenergy photons. The BSDF varies across the beam when the inhomogeneity is present due to the change in beam quality. The backscattered electrons from lead inhomogeneity have a range in the order of 5-7 mm. CONCLUSION: Higher atomic number inhomogeneities result in an increase in BSDF, as they have higher scattering cross section for the secondary electrons. The increase in dose was noticed for few millimeters upstream from the metallic inhomogeneity, which suggests that the range of backscattered electrons is very small. Since the factors affecting the BSDF at the interface are energy dependent, it is expected that the variation in BSDF will also be sensitive to the beam energy.     

Reactivation in post-traumatic chronic osteomyelitis: ultrasonographic findings.

OBJECTIVE: To describe the ultrasound (US) features of reactivation in post-traumatic chronic osteomyelitis in adults. METHODS: Twelve patients with clinical suspicion of reactivation of chronic osteomyelitis, secondary to trauma, surgery, and who were investigated with US were selected for the study. The following US features were assessed: periosseous fluid collection, bone changes (periosteal reactions, cortical irregularity, callus, sequestrum and cloaca) and soft-tissue changes (cellulitis and sinus tracts). US findings were correlated with plain radiography (n = 11), computed tomography (n = 3) and magnetic resonance imaging (n = 2). RESULTS: US detected fluid collections in all patients (12 periosseous and 3 in soft tissues), bone changes in 10 and sinus tracts and cellulitis in 5 patients each. Bone changes detected were cortical irregularity (n = 10), discontinuity of cortex (n = 7), sequestrum (n = 2), callus (n = 2), periosteal reaction (n = 1) and cloaca (n = 1). Cellulitis was seen in 5 patients and sinus tracts in 5. Reactivation was confirmed at surgery in all patients. CONCLUSION: US is a reliable noninvasive imaging modality for the diagnosis of reactivation of post-traumatic chronic osteomyelitis in adults.     

Mcm10 and And-1/CTF4 recruit DNA polymerase alpha to chromatin for initiation of DNA replication

The MCM2-7 helicase complex is loaded on DNA replication origins during the G1 phase of the cell cycle to license the origins for replication in S phase. How the initiator primase-polymerase complex, DNA polymerase alpha (pol alpha), is brought to the origins is still unclear. We show that And-1/Ctf4 (Chromosome transmission fidelity 4) interacts with Mcm10, which associates with MCM2-7, and with the p180 subunit of DNA pol alpha. And-1 is essential for DNA synthesis and the stability of p180 in mammalian cells. In Xenopus egg extracts And-1 is loaded on the chromatin after Mcm10, concurrently with DNA pol alpha, and is required for efficient DNA synthesis. Mcm10 is required for chromatin loading of And-1 and an antibody that disrupts the Mcm10-And-1 interaction interferes with the loading of And-1 and of pol alpha, inhibiting DNA synthesis. And-1/Ctf4 is therefore a new replication initiation factor that brings together the MCM2-7 helicase and the DNA pol alpha-primase complex, analogous to the linker between helicase and primase or helicase and polymerase that is seen in the bacterial replication machinery. The discovery also adds to the connection between replication initiation and sister chromatid cohesion.