Evaluation of native Trichoderma spp. against pathogens infecting small cardamom

A survey was carried out during 2008-2009 period in the high ranges of Idukki district for isolation and in vitro screening of native Trichoderma spp. against Fusarium oxysporum and Colletotrichum gloeosporioides, pathogens of small cardamom. All the 29 isolates were plated against the two selected plant pathogens in order to test their antagonistic potential and were found to be inhibitive to the growth of F. oxysporum and C. gloeosporioides. In dual cultures, out of the 29 isolates, on seventh day the isolates CT-4, CT-5, CT-10, CT-15, CT-22 and CT-23 showed above 85 per cent reduction on the growth of Fusarium and on tenth day the isolates CT-13, CT-18 and CT-23 showed above 34 per cent reduction on the growth of Colletotrichum. The biomass, spore production, the production potential of siderophore, indole acetic acid (IAA) and hydrogen cyanide (HCN) of these isolates were also assessed. Some of the isolates showed better HCN, IAA and siderophores production. This information can be employed for the exploitation of these biocontrol agents in the high ranges of Idukki district of Kerala for management of the two potential fungal pathogens of small cardamom

Mapping genomic and transcriptomic alterations spatially in epithelial cells adjacent to human breast carcinoma.

Almost all genomic studies of breast cancer have focused on well-established tumours because it is technically challenging to study the earliest mutational events occurring in human breast epithelial cells. To address this we created a unique dataset of epithelial samples ductoscopically obtained from ducts leading to breast carcinomas and matched samples from ducts on the opposite side of the nipple. Here, we demonstrate that perturbations in mRNA abundance, with increasing proximity to tumour, cannot be explained by copy number aberrations. Rather, we find a possibility of field cancerization surrounding the primary tumour by constructing a classifier that evaluates where epithelial samples were obtained relative to a tumour (cross-validated micro-averaged AUC = 0.74). We implement a spectral co-clustering algorithm to define biclusters. Relating to over-represented bicluster pathways, we further validate two genes with tissue microarrays and in vitro experiments. We highlight evidence suggesting that bicluster perturbation occurs early in tumour development

Heterogeneity of circulating tumour cell-associated genomic gains in breast cancer and its association with the host immune response.

Tumor cells that preferentially enter circulation include the precursors of metastatic cancer. Previously, we characterized circulating tumor cells (CTC) from patients with breast cancer and identified a signature of genomic regions with recurrent copy-number gains. Through FISH, we now show that these CTC-associated regions are detected within the matched untreated primary tumors of these patients (21% to 69%, median 55.5%, n = 19). Furthermore, they are more prevalent in the metastases of patients who died from breast cancer after multiple rounds of treatment (70% to 100%, median 93%, samples n = 41). Diversity indices revealed that higher spatial heterogeneity for these regions within primary tumors is associated with increased dissemination and metastasis. An identified subclone with multiple regions gained (MRG clone) was enriched in a posttreatment primary breast carcinoma as well as multiple metastatic tumors and local breast recurrences obtained at autopsy, indicative of a distinct early subclone with the capability to resist multiple lines of treatment and eventually cause death. In addition, multiplex immunofluorescence revealed that tumor heterogeneity is significantly associated with the degree of infiltration of B lymphocytes in triple-negative breast cancer, a subtype with a large immune component. Collectively, these data reveal the functional potential of genetic subclones that comprise heterogeneous primary breast carcinomas and are selected for in CTCs and posttreatment breast cancer metastases. In addition, they uncover a relationship between tumor heterogeneity and host immune response in the tumor microenvironment. SIGNIFICANCE: As breast cancers progress, they become more heterogeneous for multiple regions amplified in circulating tumor cells, and intratumoral spatial heterogeneity is associated with the immune landscape

Engraftment of Mouse Embryonic Stem Cells Differentiated by Default Leads to Neuroprotection, Behaviour Revival and Astrogliosis in Parkinsonian Rats

<div><p>We report here protection against rotenone-induced behavioural dysfunction, striatal dopamine depletion and nigral neuronal loss, following intra-striatal transplantation of neurons differentiated from murine embryonic stem cells (mES). mES maintained in serum free medium exhibited increase in neuronal, and decrease in stem cell markers by 7th and 10th days as revealed by RT-PCR and immunoblot analyses. Tyrosine hydroxylase, NURR1, PITX3, LMX1b and c-RET mRNA showed a significant higher expression in differentiated cells than in mES. Dopamine level was increased by 3-fold on 10th day as compared to 7 days differentiated cells. Severity of rotenone-induced striatal dopamine loss was attenuated, and amphetamine-induced unilateral rotations were significantly reduced in animals transplanted with 7 days differentiated cells, but not in animals that received undifferentiated ES transplant. However, the ratio of contralateral to ipsilateral swings in elevated body swing test was significantly reduced in both the transplanted groups, as compared to control. Striatal grafts exhibited the presence of tyrosine hydroxylase positive cells, and the percentage of dopaminergic neurons in the substantia nigra was also found to be higher in the ipsilateral side of 7 days and mES grafted animals. Increased expression of CD11b and IBA-1, suggested a significant contribution of these microglia-derived factors in controlling the limited survival of the grafted cells. Astrocytosis in the grafted striatum, and significant increase in the levels of glial cell line derived neurotrophic factor may have contributed to the recovery observed in the hemiparkinsonian rats following transplantation.</p></div

Changes in mRNA expression levels in cells following differentiation.

<p>Undifferentiated and differentiated embryonic stem cells (ES), and 7 days and 10 days differentiated (7 d; 10 d) cells were analysed for the mRNA expression of stem cells (NANOG; OCT3/4), neuronal (NESTIN; MAP2) and dopaminergic markers (TH; Pitx3; c-RET; NURR1; LMX1B; ENG). (A) Equal amount of cDNA was amplified for each gene and representative image of agarose gel bands obtained following electrophoresis. (B) Band intensities as normalised by hypoxanthine-guanine phosphoribosyl transferase (HGPRT; as housekeeping gene), showing relative gene expression. Results are Mean ± S.E.M, n = 3–4. *<i>p</i>≤0.05 vs. ES group, #<i>p</i>≤0.05 vs. 7 d group; Student's t-test.</p

Characterization of the grafted cells in the striatum.

<p>Striatal sections of the rotenone-infused control (ROT), embryonic stem cells transplanted animal (ES) and 7 days differentiated cells transplanted animal (7 d) were processed for presence of dopaminergic cells positive for tyrosine hydroxylase (TH) immunoreactivity. Sections were stained with cresyl violet (A, E and I) for localization of neurons (Scale bars for both inset as well as the main frame are 100 µm), and with DAPI for nuclei (B,F,J). Sham transplanted (ROT; B–D); undifferentiated embryonic stem cells transplanted (ES; F–H) and 7 days differentiated cells grafted (7 d; J–L) striatal images are shown. Scale bar is 50 µm for these figures. Scale bar for the magnified images showing TH positive cells, in the right hand corners of the lower panel is 10 µm.</p

mRNA expression of certain glial markers in the striatum.

<p>(A) Gels showing amplified cDNA bands for different primers. The cDNA was prepared from the mRNA isolated from the control and treated sides of the striatum of the embryonic stem cells, undifferentiated (ES) and 7 days differentiated (7 d) cell transplanted groups. Ratios of the band intensities for GFAP (B), GDNF (C), IBA-1 (D), CD11b-A (E) and CD11b-B (F) as normalised with hypoxanthine-guanine phosphoribosyl transferase (HGPRT) and fold change of band intensities of the treated side compared to the control side was calculated for each animal. Results are presented as Mean ± SEM. *<i>p</i>≤0.05 vs control side of the same group, ^#<i>p</i>≤0.05 vs treated side of the ES transplanted group, n = 3.</p

Co-localization of GFAP and murine specific thymocyte antigen 1.

<p>Striatal sections of 20 µm thickness of embryonic stem cells, undifferentiated (ES; A–H) and 7 days differentiated cells (7 d; I–P) transplanted group were co-stained for glial fibrillary acidic protein (GFAP; B,F,J,N) and murine specific thymocyte antigen 1 (THY1; C,G,K,O) to examine the origin of these glial cells. Figures A–D and I–L are images from locations within the graft, whereas E–H and M–P are locations along the margin of the graft, showing the host tissue and graft. The dotted lines demarcate the margin. GFAP staining is seen outside the graft margin, whereas THY1 positive cells are seen only inside the graft. Scale bar 50 µm.</p