phospholipid PIP2 and the small GTPase Cdc42 to N-WASP results in its activation.
PIP2 Cdc42 GTPase
Phosphatidylinositol 4,5-bisphosphate or PtdIns(4,5)P2, also known simply as PIP2, is a minor phospholipid component of cell membranes. PtdIns(4,5)P2 is enriched at the plasma membrane where it is an important substrate for a number of important signaling proteins.
Regulation of Cdc42 GTPase Activity in the Formation of Hyphae in Candida albicansFormula
The human fungal pathogen Candida albicans can switch between yeast, pseudohyphal, and hyphal morphologies. To investigate whether the distinctive characteristics of hyphae are due to increased activity of the Cdc42 GTPase, strains lacking negative regulators
......."induction of p53, p16Ink4a, p21Cip1, and senescence-associated β-galactosidase expressions. Furthermore, Cdc42 activation is sufficient to promote a premature cellular senescence phenotype that depends on p53. These results suggest a role of Cdc42 activity in regulating mammalian genomic stability and aging-related physiology.
We have now gotten down to at least one of the culprits.
The results suggest that BCH domains of BNIP-2 and Cdc42GAP represent a novel protein-protein interaction domain that could potentially determine and/or modify the physiological roles of these molecules.
BCH domains of BNIP-2 and Cdc42GAP seems like it is being examined................. ================
............."Current work is aimed at addressing the detailed intracellular localization of BNIP-2.
In conclusion, our present work has shown that BCH domains of BNIP-2 and Cdc42GAP define a novel class of protein-protein interaction domain that includes various uncharacterized proteins. It may represent another example of proteins that form dimers as a functional necessity such as: various receptor tyrosine kinases (40), STAT transcription factors (41), c-Raf (42, 43), and various members of the Bcl family (44, 45). Our work also highlights the fact that although several structural studies have used the catalytic domain of Cdc42GAP to define a precise interaction with Cdc42 (and Rho) (18, 46) there is still much to be understood about how this protein is targeted and activated and indeed what its actual physiological role is. A better understanding of the structure and functional roles of the BCH domains of BNIP-2, Cdc42GAP, and of other proteins will answer these questions. "................
Formic acid (systematically called methanoic acid) is the simplest carboxylic acid. Its formula is HCOOH or HCO2H. It is an important intermediate in chemical synthesis and occurs naturally, most notably in the venom of bee and ant stings. In fact, its name comes from the Latin word for ant, formica, referring to its early isolation by the distillation of ant bodies. Esters, salts and the anion derived from formic acid are referred to as formate.
Changing from O2 to HCOOH or HCO2H. chemical synthesis esters salts anion formate
make the formins.
Formins (formin homology proteins) are a group of proteins that are involved in the polymerisation of actin and associate with the fast-growing end (barbed end) of actin filaments . The structure of formins is mainly conserved in the C-terminal Formin Homology (FH) domains namely FH1, FH2 and the more loosely defined FH3 domain . The actin nucleation-promoting activity of S. cerevisiae formins has been localized to the FH1-FH2 domains. Other commonly shared domains are a GTPase binding domain (GBD) that bind for example Cdc42 at the N-terminus, and a DAD or Dia-autoregulatory domain, toward the C-terminus.
"The formins: active scaffolds that remodel the cytoskeleton"
Unleashing formins to remodel the actin and microtubule cytoskeletons
Formins are highly conserved proteins that have essential roles in remodelling the actin and microtubule cytoskeletons to influence eukaryotic cell shape and behaviour. Recent work has identified numerous cellular factors that locally recruit, activate or inactivate formins to bridle and unleash their potent effects on actin nucleation and elongation. The effects of formins on microtubules have also begun to be described, which places formins in a prime position to coordinate actin and microtubule dynamics. The emerging complexity in the mechanisms governing formins mirrors the wide range of essential functions that they perform in cell motility, cell division and cell and tissue morphogenesis.
RhoA-Dependent Vascular Smooth Muscle Cell–Specific Transcription Adding Diaphanous Formins to the Puzzle
RhoA-dependent vascular smooth muscle–specific transcription. Many microenvironmental cues, including growth factors, cell–cell contacts, and extracellular matrix, induce activation of RhoA. ROK, PKN, and Dia1/2 are the RhoA effector molecules that are operating in SMCs to regulate cytoskeletal reorganization and nuclear translocation of MRTF-A/B. These transcriptional factors then facilitate the binding of SRF to one or more CArG sequences in the promoter of many SMC-specific genes. CArG indicates CC(AT)6GG sequences; Dia, diaphanous formin; MRTF, myocardin-related transcriptional factor; PKN, protein kinase N; ROK, RhoA-binding kinase; SRF, serum-response factor.
Diaphanous 1 and 2 Regulate Smooth Muscle Cell Differentiation by Activating the Myocardin-Related Transcription Factors
Objective— We have previously shown that smooth muscle cell (SMC) differentiation marker gene expression is regulated by the small GTPase, RhoA. The objective of the present study was to determine the contributions of the RhoA effectors, diaphanous 1 and 2 (mDia1 and mDia2), to this regulatory mechanism.
BIOLOGICAL OVERVIEW The diaphanous (dia) locus plays a critical role during cytokinesis: various combinations of dia mutations result in multinucleate spermatids, polyploid larval neuroblasts and adult follicle cells, and pupal lethality (Castrillon and Wasserman, 1994)
Dual roles of myocardin-related transcription factors in epithelial–mesenchymal transition via slug induction and actin remodeling
MRTFs also increase the expression levels of actin cytoskeletal proteins via serum response factor, thereby triggering reorganization of the actin cytoskeleton. Thus, MRTFs are important mediators of TGF-β1–induced EMT.
;..........."MRTFs form complexes with Smad3. Via Smad3, the MRTF–Smad3 complexes bind to a newly identified cis-element GCCG-like motif in the promoter region of Canis familiaris and the human slug gene, which activates slug transcription and thereby dissociation of cell–cell contacts''''''''''''
Abbreviations used in this paper: α-SMA, α–smooth muscle actin; CA, constitutive active; ChIP, chromatin immunoprecipitation; DN, dominant negative; EMT, epithelial–mesenchymal transition; HMGA2, high mobility group A2; LMB, leptomycin B; MRTF, myocardin-related transcription factor; SBE, Smad-binding element; SRF, serum response factor.
Protein Kinase N (PKN) and PKN-Related Protein Rhophilin as Targets of Small GTPase Rho
The Rho guanosine 5'-triphosphatase (GTPase) cycles between the active guanosine triphosphate (GTP)-bound form and the inactive guanosine diphosphate-bound form and regulates cell adhesion and cytokinesis, but how it exerts these actions is unknown. The yeast two-hybrid system was used to clone a complementary DNA for a protein (designated Rhophilin) that specifically bound to GTP-Rho. The Rho-binding domain of this protein has 40 percent identity with a putative regulatory domain of a protein kinase, PKN. PKN itself bound to GTP-Rho and was activated by this binding both in vitro and in vivo. This study indicates that a serine-threonine protein kinase is a Rho effector and presents an amino acid sequence motif for binding to GTP-Rho that may be shared by a family of Rho target proteins.
"The RhoA-binding protein, Rhophilin-2, Regulates Actin Cytoskeleton Organization*
From the work presented here, we propose that the two human Rhophilin members, Rhophilin-1 and Rhophilin-2, have different activities. Although the exact normal function of Rhophilin-2 is not clear, one function of Rhophilin-2 may be to limit stress fiber formation in the absence of high levels of activated RhoA. Alternatively, Rhophilin-2 may normally be activated by RhoA and promote actin stress fiber disassembly, which may be requiredin vivo for biological processes such as cell motility and cytokinesis and whose activity may be normally masked by the action of other effector proteins (e.g. ROCK and Dia) that promote stress fiber assembly. Future studies are aimed at identifying additional protein targets in the cell responsible for mediating Rhophilin-2-induced cytoskeletal changes to clarify its normal function.....
---------- Phosphatidylinositol 4-phosphate 5-kinase (PIP-5kin) regulates actin cytoskeletal reorganization through its product phosphatidylinositol 4,5-bisphosphate.
Rho-associated kinases termed ROCK, Rho-kinase, and ROK mediate Rho-induced actin cytoskeletal reorganization (13-15) and subsequent repulsive responses of neurites including growth cone collapse and neurite retraction. Evidence for this includes the findings that overexpression of ROCK as well as the active mutant of RhoA, RhoAV14, inhibits neurite formation in N1E-115 cells (16, 17). In addition, pharmacological inactivation of endogenous ROCK, similar to inhibition of Rho activity, interferes with LPA-induced neurite retraction in N1E-115 cells (16, 17) and Ephrin-A5-induced growth cone collapse in retinal neurons (12).
Exchange of a Single Amino Acid Switches the Substrate Properties of RhoA and RhoD toward Glucosylating and Transglutaminating Toxins*
Rho GTPases are the preferred targets of various bacterial cytotoxins, including Clostridium difficile toxins A and B, Clostridium sordellii lethal toxin, the cytotoxic necrotizing factors (CNF1) from Escherichia coli, and the dermonecrotizing toxin (DNT) from Bordetella species. The toxins inactivate or activate specific sets of Rho GTPases by mono-O-glucosylation and deamidation/transglutamination, respectively. Here we studied the structural basis of the recognition of RhoA, which is modified by toxin B, CNF1, and DNT, in comparison with RhoD, which is solely a substrate for lethal toxin. We found that a single amino acid residue in RhoA and RhoD defines the substrate specificity for toxin B and lethal toxin. Change of serine 73 to phenylalanine in RhoA turned RhoA into a substrate for lethal toxin. Accordingly, change of the equivalently positioned phenylalanine 85 in RhoD with serine allowed glucosylation by toxin B. Comparable results were achieved with the Rho-activating and transglutaminating enzymes CNF1 and DNT. Here, amino acid glutamate 64 of RhoA and the equivalent aspartate 76 of RhoD define substrate specificity for CNF1 and DNT, respectively. These data indicate that single amino acid residues located in the switch II region of Rho proteins determine enzyme specificity for diverse bacterial toxins.
transglutaminating enzymes CNF1 and DNT.
Is TC 10 the link to the genes in the toxin of BTV (T2 nano virus)?
Some bacterial protein toxins alter the activity of Rho/Ras GTPases in a highly specific manner. Clostridial glycosylating toxins like toxin B of Clostridium difficile and Clostridium sordellii lethal toxin mono-glucosylate low molecular mass GTPases and inhibit their function (11); other toxins like Escherichia coli cytotoxic necrotizing factor 1 (CNF1)2 and Bordetella dermonecrotic toxin (DNT) activate Rho GTPases (12) (see Fig. 1). The glucosylating toxins modify a conserved threonine residue within the switch I region (position 37 in RhoA) that is essential for nucleotide binding (13-15). Toxin B modifies the Rho family members RhoA, -B, -C, Rac1, Cdc42, RhoG, and TC10 (14, 16).
...."Constructing RhoA-Rac1 chimeras, Müller and coworkers (34) found that amino acids 22-27 in Rac1 were of major impact for recognition by lethal toxin of C. sordellii strain 1522 and toxin B from C. difficile strains 1470 and 8864."...........
we identified the histone 3 lysine 9 (H3K9)-specific demethylase, jmjd1a. GST pull-down assays demonstrated that jmjd1a bound all 3 myocardin family members, and further mapping studies showed that the jumonjiC domain of jmjd1a was sufficient to mediate this interaction. Overexpression of jmjd1a in multipotential 10T1/2 cells decreased global levels of di-methyl H3K9, stimulated the SM alpha-actin and SM22 promoters, and synergistically enhanced MRTF-A- and myocardin-dependent transactivation
Jmjd1a and Jmjd2c histone H3 Lys 9 demethylases regulate self-renewal in embryonic stem cells
Embryonic stem (ES) cells are pluripotent cells with the ability to self-renew indefinitely. These unique properties are controlled by genetic factors and chromatin structure. The exit from the self-renewing state is accompanied by changes in epigenetic chromatin modifications such as an induction in the silencing-associated histone H3 Lys 9 dimethylation and trimethylation (H3K9Me2/Me3) marks. Here, we show that the H3K9Me2 and H3K9Me3 demethylase genes, Jmjd1a and Jmjd2c, are positively regulated by the ES cell transcription factor Oct4. Interestingly, Jmjd1a or Jmjd2c depletion leads to ES cell differentiation, which is accompanied by a reduction in the expression of ES cell-specific genes and an induction of lineage marker genes. Jmjd1a demethylates H3K9Me2 at the promoter regions of Tcl1, Tcfcp2l1, and Zfp57 and positively regulates the expression of these pluripotency-associated genes. Jmjd2c acts as a positive regulator for Nanog, which encodes for a key transcription factor for self-renewal in ES cells. We further demonstrate that Jmjd2c is required to reverse the H3K9Me3 marks at the Nanog promoter region and consequently prevents transcriptional repressors HP1 and KAP1 from binding. Our results connect the ES cell transcription circuitry to chromatin modulation through H3K9 demethylation in pluripotent cells. Keywords: Histone demethylase, embryonic stem cell, chromatin immunoprecipitation, self-renewal, pluripotency, Jumonji
"RhoA effectors, diaphanous 1 and 2 (mDia1 and mDia2)"
We now report that POPX2 interacts with the formin protein mDia1 (DIAPH1). This interaction is enhanced when mDia1 is activated by RhoA. The binding of POPX2 to mDia1 or to an mDia-containing complex greatly decreases the ability of mDia1 to activate transcription from the SRE. We propose that the interaction between mDia1 and POPX2 (PPM1F) serves to regulate both the actin cytoskeleton and SRF-mediated transcription, and to link the CDC42/RAC1 pathways with those of RhoA.
"n conclusion, POPX2 can be considered not only as a phosphatase in the context of targeting phospho-proteins, but also as a scaffolding protein via its N-terminal non-catalytic domain. Because of its link to both CDC42/RAC1 pathways [via PIX-PAK (Koh et al., 2002Go) and RhoA pathways (via mDia1, this report)], POPX2 represents an important player in cross-talk between the various Rho GTPase signalling pathways that underlie diverse activities; including morphological changes, transcription and the cell cycle.""
Zinc finger protein SNAI2 is a protein that in humans is encoded by the SNAI2 gene.
This gene encodes a member of the Snail family of C2H2-type zinc finger transcription factors. The encoded protein acts as a transcriptional repressor that binds to E-box motifs and is also likely to repress E-cadherin transcription in breast carcinoma. This protein is involved in epithelial-mesenchymal transitions and has antiapoptotic activity. Mutations in this gene may be associated with sporadic cases of neural tube defects.".........
Note defect in melanocyte development; one of the first genetic disorders for which a pedigree was presented in 1786 Inheritance autosomal dominant; frequency is about 2.5/105 newborns # Phenotype and clinics congenital patches of white skin and white hair, principally located on the scalp, forehead, chest and abdomen and on the limbs; several patients report lifelong severe constipation; a hierarchical correlation has been elaborated between severe or mild phenotypic traits and the associated KIT mutations; in a few patients with interstitial deletions mental retardation and congenital anomalies have been also described # etiology :defective melanoblasts proliferation, survival and migration from the neural crest during development and defective migration of enteric-plexus ganglion cells from the neural crest to the gut # pathology : white spotting in human piebaldism results from the absence of melanocytes from the nonpigmented patches of skin and from hairbulbs in the white patches of hair; occasionally, individuals lack ganglion cells of the intestinal enteric neural plexus, which like melanoblasts, are derived from the neural crest Neoplastic risk an increased risk of epithelioma has been reported Prognosis in contrast to vitiligo, piebaldism is both congenital and non-progressive Inborn conditions a few patients with interstitial deletions of chromosome 4q12-q21.1 have been identified; they are charaterized by multiple congenital anomalies, short stature and mental retardation.
KIT Location in 4q12 DNA/RNA Description 21 exons Protein Description transmembrane SCF/MGF receptor with tyrosine kinase activity; binding of ligand (SCF) induces receptor dimerization, autophosphorylation and signal transduction via molecules containing SH2- domains Mutations Note see diagram: Loss-of-function mutations # Germinal loss of function mutations resulting in haploinsufficiency of the receptor; different kinds of point mutations have been identified (diagram): missense substitutions (Glu583Lys; Phe584Leu; Ala621Thr; His650Pro; Gly664Arg; Gly791Arg; Arg796Gly; Val812Gly; Glu861Ala) and small deletions (641del2; 892 del12) in the intracellular tyrosine kinase domain; correlate with severe piebald phenotypes, because of dominant-negative inhibition of the KIT receptor via formation of impaired receptor heterodimers between a normal and a mutant KIT monomer, and a 75% decrease of KIT- dependent signal transduction. # proximal frameshifts (84del1; 249del4); Trp557Term; and missense mutations (Cys136Arg; Ala178Thr; Met318Gly) associated with a mild piebald phenotype, the result of pure haploinsufficiency due to a 50% decrease of KIT-dependent signal transduction # distal frameshifts: 630insA; and splice junction mutations (IVS1+4G-A; IVS12+1G- A), located near the intracellular TK domain associated with variable phenotypes, as the truncated polypeptides via incorporation into nonfunctional receptor heterodimers would decrease KIT-dependent signal transduction by 50-75%, depending on their stability # complete deletions of the entire KIT gene ("null" mutations) result in a mild- intermediate phenotype.
Gene Name PDGFRA Location 4q12 Note is also deleted in patients with interstitial cytogenetic deletions (contiguous gene syndrome)
Gene Name SCF/MGF Location 12q22 Note no alteration of this gene has been so far identified in typical patients; at difference with the mouse system, where "steel" mice bearing SCF mutations show the "white spotting" phenotype likewise W mice bearing kit mutations; however, as mutations of KIT could not be detected in a consistent fraction of these patients, involvement of SCF is still an open question.
OMIM 1472800 Orphanet Piebaldism HGMD 120117
Novel mutations and deletions of the KIT (steel factor receptor) gene in human piebaldism. Ezoe K, Holmes SA, Ho L, Bennett CP, Bolognia JL, Brueton L, Burn J, Falabella R, Gatto EM, Ishii N American journal of human genetics. 1995 ; 56 (1) : 58-66. PMID 7529964 A 12-bp deletion (7818del12) in the c-kit protooncogene in a large Italian kindred with piebaldism. Riva P, Milani N, Gandolfi P, Larizza L Human mutation. 1995 ; 6 (4) : 343-345. PMID 8680409
Mutations in the ligand-binding domain of the kit receptor: an uncommon site in human piebaldism. Fleischman RA, Gallardo T, Mi X The Journal of investigative dermatology. 1996 ; 107 (5) : 703-706. PMID 8875953 Piebaldism with deafness: molecular evidence for an expanded syndrome. Spritz RA, Beighton P American journal of medical genetics. 1998 ; 75 (1) : 101-103. PMID 9450866
REVIEW articles automatic search in PubMed Last year publications automatic search in PubMed
Written 09-1998 Lidia Larizza and Alessandro Beghini
Updated 06-2000 Lidia Larizza and Alessandro Beghini
Piebaldism is due to an absence of melanocytes in affected skin and hair follicles as a result of mutations of the KIT proto-oncogene.1 As of a 2001 review by Richards et al, 14 point mutations, 9 deletions, 2 nucleotide splice mutations, and 3 insertions of the KIT gene were believed to be mutations causing piebaldism.2 The severity of phenotypic expression in piebaldism correlates with the site of the mutation within the KIT gene. The most severe mutations seem to be dominant negative missense mutations of the intracellular tyrosine kinase domain, whereas mild piebaldism appears related to mutations occurring in the amino terminal extracellular ligand-binding domain with resultant haplo insufficiency.
14 point mutations 9 deletions 2 nucleotide splice mutations, and 3 insertions of the KIT gene were believed to be mutations causing piebaldism
The most severe mutations seem to be dominant negative missense mutations of the intracellular tyrosine kinase domain, whereas mild piebaldism appears related to mutations occurring in the amino terminal extracellular ligand-binding domain with resultant haplo insufficiency.
...". Intriguingly, contactin can be isolated in a complex with the intracellular src family tyrosine kinase fyn ( Zisch et al. 1995). The mechanism of this association is unknown, but likely requires an intermediate membrane-spanning component."............
..."the association of fyn with contactin may be central to contactin signaling. Several protein tyrosine phosphatases (PTPs) can activate src family kinases in various systems, suggesting a PTP to be a candidate component of a contactin signaling complex.".......
...."PTPα also associates with, dephosphorylates, and activates brain fyn ..., a src family kinase that plays a role in axonal growth, myelination, spatial learning and memory "......
...."The pXJ41 vectors expressing PTPα, VSVG-tagged PTPα, fyn, and CD45 have been described "...........
I just read where childhood sarcomas are rising. Lipomas are what was labeled on me about 10 years ago, the soft tissue under the skin, when it was removed.
What do we know about the formation or sarcomas, lipomas, myomas, fibromas.
The all have mas(s) in common.
A sarcoma (from the Greek 'sarx' meaning "flesh") is a general term describing a malignant neoplasm, or cancer, that arises from transformed connective tissue cells, such as bone, cartilage, and fat cells, which originate from embryonic mesoderm.
This is in contrast to carcinomas, which are of epithelial origin (breast, colon, pancreas, and others).....
........."The term soft tissue sarcoma is used to describe tumors of soft tissue, which includes elements that are in connective tissue, but not derived from it (such as muscles and blood vessels).".......
Sarcomas are given a number of different names, based on the type of tissue from which they arise. For example, osteosarcoma arises from bone, chondrosarcoma arises from cartilage, and leiomyosarcoma arises from smooth muscle.".............
This was the kind that children are getting:
Sarcoma botryoides or botryoid sarcoma or botryoid rhabdomyosarcoma is a subtype of embryonal rhabdomyosarcoma, that can be observed in the walls of hollow, mucosa lined structures such as the nasopharynx, common bile duct, urinary bladder of infants and young children or the v spot in females, typically younger than age 8. The name comes from the gross appearance of "grape bunches" (botryoid in Greek).
Under the microscope one can see rhabdomyoblasts that may contain cross-striations. Tumor cells are crowded in a distinct layer beneath the v spotl epithelium ( cambium layer)."....
Definition By Mayo Clinic staff CLICK TO ENLARGE Image of a lipoma Lipoma
A lipoma is a slow-growing, fatty lump that's most often situated between your skin and the underlying muscle layer. Often a lipoma is easy to identify because it moves readily with slight finger pressure. It's doughy to touch and usually not tender. You may have more than one lipoma. Lipomas can occur at any age, but they're most often detected during middle age.
A lipoma isn't cancer and is usually harmless. Treatment generally isn't necessary, but if the lipoma is in a location that bothers you, is painful or is growing, you may want to have it removed.
A lipoma is a growth of fat cells in a thin, fibrous capsule usually found just below the skin. Lipomas are found most often on the torso, neck, upper thighs, upper arms, and armpits, but they can occur almost anywhere in the body. One or more lipomas may be present at the same time. Lipomas are the most common noncancerous soft tissue growth.
What causes a lipoma?
The cause of lipomas is not completely understood, but the tendency to develop them is inherited. A minor injury may trigger the growth. Being overweight does not cause lipomas.
A leiomyoma (plural is 'leiomyomata') is a benign smooth muscle neoplasm that is not premalignant. They can occur in any organ, but the most common forms occur in the uterus, small bowel and the esophagus.
* Greek: o leios = smooth o muV = (myo) mouse or muscle o oma = tumor * Latin: o Fibra = fiber
Fibromas (or fibroid tumors or fibroids) are benign tumors that are composed of fibrous or connective tissue. They can grow in all organs, arising from mesenchyme tissue. The term "fibroblastic" or "fibromatous" is used to describe tumors of the fibrous connective tissue. When the term fibroma is used without modifier, it is usually considered benign, with the term fibrosarcoma reserved for malignant tumors.[/b]
We report the clinical and histologic features of 2 cutaneous tumors demonstrating many of the usual features of sclerotic fibroma, an entity that is well described in the literature. Our cases differed from the usual form of sclerotic fibroma in that they demonstrated marked focal cellular pleomorphism without increased mitotic activity. We suggest the term pleomorphic sclerotic fibroma for these lesions.
Hemangiopericytoma was first described by Stout and Murray as a less organoid form of glomus tumour arising from pericytes.(Hemangiopericytoma: a vascular tumour featuring Zimmermann's pericytes. Ann Surg 1942: 116: 26-33).
Enzinger and Smith (Hemangiopericytoma. An analysis of 106 cases.Hum Pathol. 1976 Jan;7(1):61-82.) divided this tumour into two groups: I. Adult II. Infantile or congenital hemangiopericytoma.
The concept of 'Hemangiopericytoma' has been questioned my many authors due to absence of reproducible diagnostic criteria.
(Hemangiopericytoma: a dying breed? Reappraisal of an ‘entity’ and its variants: a hypothesis. Curr Diagn Pathol 1994;1:19–23.)
Many soft tissue tumours focally display 'hemangiopericytoma- like' pattern.
Usually present at birth or noted soon after. This tumour has a benign course.
Site: Mostly solitary lesion, located on the head and neck area, extremities or the trunk.
Microscopic features: The multilobulated tumour displays polymorphic population of cells consisting of spindle cells with myofibroblastic features and a more primitive round cells (both are smooth muscle actin positive).
There are distinct intravascular and perivascular satellite nodules outside the main tumour mass. There is evidence of intravascular endothelial cell proliferation.
Mitotic figures are noted together with focal necrosis and vascular invasion (this is not related to bad prognosis).
It has been suggested that infantile myofibromatosis and infantile hemangiopericytoma represent different stages of maturation of the same lesion.
Infantile hemangiopericytoma versus infantile myofibromatosis. Study of a series suggesting a continuous spectrum of infantile myofibroblastic lesions. Am J Surg Pathol. 1994;18(9):922-30.
Hemangiopericytoma in pediatric ages: a report from the Italian and German Soft Tissue Sarcoma Cooperative Group.Cancer. 2001;92(10):2692-8.
Hemangiopericytoma in children and infants. Cancer 2000 ;88(1):198-204
Myopericytoma include a range of lesions displaying features ranging from myofibromatosis to those resembling glomus tumour with hemangiopericytoma-like blood vessels.
Microscopic features: The tumour exhibits biphasic pattern characterized by primitive spindle cell with hemangiopericytoma-like vascular pattern and fascicles of spindle cells with eosinophilic cytoplasm, resembling smooth muscle.
Glomangiopericytoma is characterized by branching blood vessels lined by endothelial cells surrounded by epithelioid cells (glomoid appearance).
-Malignant myopericytoma: expanding the spectrum of tumours with myopericytic differentiation. Histopathology. 2002;41(5):450-60.
=================== (WO/2007/107380) USE OF POL III PROMOTERS FOR CONTROLLED EXPRESSION OF THERAPEUTIC PROTEINS
Genes, which are carried on chromosomes, are the basic physical and functional units of heredity. Genes are specific sequences of bases that encode instructions on how to make proteins. Although genes get a lot of attention, it's the proteins that perform most life functions and even make up the majority of cellular structures. When genes are altered so that the encoded proteins are unable to carry out their normal functions, genetic disorders can result.
A normal gene may be inserted into a nonspecific location within the genome to replace a nonfunctional gene. This approach is most common.
- An abnormal gene could be swapped for a normal gene through homologous recombination. - The abnormal gene could be repaired through selective reverse mutation, which returns the gene to its normal function. - The regulation (the degree to which a gene is turned on or off) of a particular gene could be altered.
In most gene therapy studies, a "normal" gene is inserted into the genome to replace an "abnormal," disease-causing gene. A carrier molecule called a vector must be used to deliver the therapeutic gene to the patient's target cells. Currently, the most common vector is a virus that has been genetically altered to carry normal human DNA. Viruses have evolved a way of encapsulating and delivering their genes to human cells in a pathogenic manner. Scientists have tried to take advantage of this capabil- ity and manipulate the virus genome to remove disease- causing genes and insert therapeutic genes.
Target cells such as the patient's liver, bone marrow, T stem- or lung cells are infected with the viral vector. The vector then unloads its genetic material containing the therapeutic human gene into the target cell. The generation of a functional protein product from the therapeutic gene restores the target cell to a normal state.
Some of the different types of viruses used as gene therapy vectors:
- Retroviruses - A class of viruses that can create dou- ble-stranded DNA copies of their RNA genomes. These copies of its genome can be integrated into the chromosomes of host cells. Human immunodeficiency virus (HIV) is a retrovirus .
- Adenoviruses - A class of viruses with double-stranded DNA genomes that cause respiratory, intestinal, and eye infections in humans. The virus that causes the common cold is an adenovirus.
- Adeno-associated viruses - A class of small, single- stranded DNA viruses that can insert their genetic material at a specific site on chromosome 19.
- Herpes simplex viruses - A class of double-stranded DNA viruses that infect a particular cell type, neurons. Her¬ pes simplex virus type 1 is a common human pathogen that causes cold sores.
Besides virus-mediated gene-delivery systems, there are several nonviral options for gene delivery. The simplest method is the direct introduction of therapeutic DNA into target cells. This approach is limited in its application because it can be used only with certain tissues and requires large amounts of DNA.
Another nonviral approach involves the creation of an artificial lipid sphere with an aqueous core. This liposome, which carries the therapeutic DNA, is capable of passing the DNA through the target cell's membrane.
Therapeutic DNA also can get inside target cells by chemically linking the DNA to a molecule that will bind to special cell receptors. Once bound to these receptors, the therapeutic DNA constructs are engulfed by the cell membrane and passed into the interior of the target cell. This delivery system tends to be less effective than other options.
Within the framework of gene therapy, viral vectors are well known as being efficient vehicles for the stable introduction of genes into human cells. The integration of the gene which is to be introduced occurs at points of the genome chosen at random and by means of well defined insertion sequences (so-called LTRs - "long terminal repeat") . The insertion cassettes carry the gene which is to be introduced and which is expressed under the control of RNAPoI II promoters, according to the biologic systems. Clinical studies with SCID-Xl patients (sever combined immunodeficiency type Xl) showed that the random insertion of the vectors into the genome can lead to an unwished activation of adjacent genes including protoon- cogenes, which results in malign diseases, in the present case in leukemia. A great number of different - although to date unsuccessful - efforts have been made ever since to optimize such viral vectors.
The expression of proteins in eucaryotes occurs through a complex system of RNA and protein factors which catalyze in a very strictly ordered way both the transcription of genomic DNA to mRNA and the translation of mRNA to proteins. Moreover, the single steps of the gene expression take place in different compartments of the cell.
The cell is equipped with separate expression systems for the expression of RNAs which are not ( ! ) translated to proteins. Especially responsible for this are the RNA polymerases (RNAPoI) I and III. The RNAPoI II transcribes the mRNA which is fundamental for protein expression.
The functional distribution to different RNA polymerases is generally very strictly regulated.
The RNA polymerase III (pol III) promoter is one found in DNA encoding 5S, U6, adenovirus VAl, Vault, telomerase RNA, tRNA genes, etc., and is transcribed by RNA polymerase III (for a review see Geiduschek and Tocchini- Valentini, 1988 Annu. Rev. Biochem. 57, 873-914; Willis, 1993 Eur. J. Biochem. 212, 1-11) . There are three major types of pol III promoters: types 1, 2 and 3 (Geiduschek and Tocchini-Valentini, 1988 supra; Willis, 1993 supra) . Type 1 pol III promoter consists of three cis-acting sequence elements downstream of the transcriptional start site a) 51 sequence element; b) an intermediate sequence element; c) 31 sequence element. 5S ribosomal RNA genes are transcribed using the type 1 pol III promoter (Specht et al., 1991 Nucleic Acids Res. 19, 2189-2191).
Eukaryotic nuclear gene expression is performed by three molecular complexes. These complexes are referred to as RNA polymerases (pol) I, II and III. Pol I transcribes the rRNA genes. Pol II transcribes the protein -encoding genes and many small nuclear RNA genes. Pol III, which is subdivided into three types, transcribes many untranslated gene products that are involved in RNA processing including 5S rRNA, tRNA, U6 RNA and Hl RNA molecules. Type I and II pol III promoters reguire intragenic sequence elements (downstream of +1) to transcribe 5S rRNA and tRNA respectively.
Type III pol III promoters requires no intragenic sequence elements for transcribing their genes. Pol III promoters transcribe short RNA that rarely exceed 300 nucleotides (nt) in length all of these transcripts terminates tightly once a run of 5 or more thymidines (T) is present. Type III Pol III promoters are now used for expressing short inhibitory RNA (siRNA) for the purpose of RNA interference. Variations of this type of construct are available in which siRNA molecules are expressed from plasmid, adenoviral or retroviral backbones.
"Variations of this type of construct are available in which siRNA molecules are expressed from plasmid, adenoviral or retroviral backbones.".............
A universal plasmid library encoding all permutations of small interfering RNA
Small interfering RNA (siRNA) is normally designed to silence preselected known genes. Such selections are inevitably prone to bias as a result of limited knowledge about the biological process, transcript identity, and functions. A library that contains all permutations of siRNA could avoid such problems. In this paper, it is shown that 5 × 107 siRNA-encoding plasmids can be constructed in a single tube by using vectors with two mutated RNA polymerase III promoters arranged in a convergent manner. Such a library was used to carry out genomewide screening of functional genes in a phenotype-driven manner. Multiple siRNAs that induce a significant increase of cell proliferation speed were identified.
Keywords: random targeting, screening, vector, dual promoters
"To overcome these limitations, the concept of siRNA libraries has been explored. Here we present the construction of siRNA libraries that contain all permutations of siRNA sequences. For this purpose we created a plasmid vector system that contains two convergent RNA polymerase III (Pol III) promoters to drive the expression of both strands of the siRNA from a single randomized region in living cells. We further demonstrated the usefulness of this type of siRNA library for phenotype-driven genomewide screening in a cell proliferation model."
DNA Polymerase I (or Pol I) is an enzyme that participates in the process of DNA replication in prokaryotes. It is composed of 928 amino acids, and is an example of a processive enzyme - it can sequentially catalyze multiple polymerisations. Discovered by Arthur Kornberg in 1956, it was the first known DNA polymerase (and, indeed, the first known of any kind of polymerase). It was initially characterized in E. coli, although it is ubiquitous in prokaryotes. In E. coli and many other bacteria, the gene which encodes Pol I is known as polA.
Pol I DNA Polymerase I (or Pol I) is an enzyme that participates in the process of DNA replication in prokaryotes. It is composed of 928 amino acids, and is an example of a processive enzyme - it can sequentially catalyze multiple polymerisations. Discovered by Arthur Kornberg in 1956, it was the first known DNA polymerase (and, indeed, the first known of any kind of polymerase). It was initially characterized in E. coli, although it is ubiquitous in prokaryotes. In E. coli and many other bacteria, the gene which encodes Pol I is known as polA.
Pol I possesses three enzymatic activities:
1. A 5' -> 3' (forward) DNA polymerase activity, requiring a 3' primer site and a template strand 2. A 3' -> 5' (reverse) exonuclease activity that mediates proofreading 3. A 5' -> 3' (forward) exonuclease activity mediating nick translation during DNA repair.
In the replication process, DNA Polymerase I removes the RNA primer (created by Primase) from the lagging strand and fills in the necessary nucleotides of the Okazaki fragments (see DNA replication) in 5' -> 3' direction, proofreading for mistakes as it goes. It is a template-dependent enzyme - it only adds nucleotides that correctly base pair with an existing DNA strand acting as a template. Ligase then joins the various fragments together into a continuous strand of DNA.
DNA polymerase I obtained from E. coli is used extensively for molecular biology research. However, the 5' -> 3' exonuclease activity makes it unsuitable for many applications. Fortunately this undesirable enzymatic activity can be simply removed from the holoenzyme to leave a useful molecule called the Klenow fragment, widely used in molecular biology. Exposure of DNA polymerase I to the protease subtilisin cleaves the molecule into a smaller fragment, which retains only the DNA polymerase and proofreading activities."............
---------- Exposure of DNA polymerase I to the protease subtilisin
Some enzymes do not need any additional components to show full activity. However, others require non-protein molecules called cofactors to be bound for activity. Cofactors can be either inorganic (e.g., metal ions and iron-sulfur clusters) or organic compounds (e.g., flavin and heme).Organic cofactors can be either prosthetic groups, which are tightly bound to an enzyme, or coenzymes, which are released from the enzyme's active site during the reaction.Coenzymes include NADH, NADPH and adenosine triphosphate. These molecules transfer chemical groups between enzymes.
An example of an enzyme that contains a cofactor is carbonic anhydrase, and is shown in the ribbon diagram above with a zinc cofactor bound as part of its active site. These tightly bound molecules are usually found in the active site and are involved in catalysis. For example, flavin and heme cofactors are often involved in redox reactions.
Enzymes that require a cofactor but do not have one bound are called apoenzymes or apoproteins. An apoenzyme together with its cofactor(s) is called a holoenzyme (this is the active form). Most cofactors are not covalently attached to an enzyme, but are very tightly bound. However, organic prosthetic groups can be covalently bound (e.g., thiamine pyrophosphate in the enzyme pyruvate dehydrogenase). The term "holoenzyme" can also be applied to enzymes that contain multiple protein subunits, such as the DNA polymerases; here the holoenzyme is the complete complex containing all the subunits needed for activity.
Coenzymes Space-filling model of the coenzyme NADH
Coenzymes are small organic molecules that transport chemical groups from one enzyme to another. Some of these chemicals such as riboflavin, thiamine and folic acid are vitamins (compounds which cannot be synthesized by the body and must be acquired from the diet). The chemical groups carried include the hydride ion (H-) carried by NAD or NADP+, the phosphate group carried by adenosine triphosphate, the acetyl group carried by coenzyme A, formyl, methenyl or methyl groups carried by folic acid and the methyl group carried by S-adenosylmethionine.
Since coenzymes are chemically changed as a consequence of enzyme action, it is useful to consider coenzymes to be a special class of substrates, or second substrates, which are common to many different enzymes. For example, about 700 enzymes are known to use the coenzyme NADH.
Coenzymes are usually continuously regenerated and their concentrations maintained at a steady level inside the cell: for example, NADPH is regenerated through the pentose phosphate pathway and S-adenosylmethionine by methionine adenosyltransferase. This continuous regeneration means that even small amounts of coenzymes are used very intensively. For example, the human body turns over its own weight in ATP each day.
An artificial enzyme is a synthetic, organic molecule prepared to recreate the active site of an enzyme.
Enzyme catalysis of chemical reactions occur with high selectivity and rate in a small part of the enzyme macromolecule known as the active site. There, the binding of a substrate close to functional groups in the enzyme causes catalysis by so-called proximity effects. It is therefore possible to create similar catalysts from small molecule mimics of enzyme active sites by combining, in a small molecule, the ability to bind substrate with catalytic functional groups. Since the artificial enzymes need to bind molecules, they are made based on a host-molecule such as a cyclodextrin, crown ethers or calixarene etc.[/u]
A number of artificial enzymes have been reported catalysing various reactions with rate increases up to 103; this is nevertheless substantially lower than natural enzymes that typically causes rate increases above 106. One of the pioneers in artificial enzyme research is chemist Ronald Breslow.[/u]
New approaches based on amino acids or peptides as characteristic molecular moieties have led to a significant expansion of the field of artificial enzymes or enzyme mimics. For instance, recent results by the group of Rob Liskamp have shown that scaffolded histidine residues can be used as mimics of certain metalloproteins and -enzymes. Especially the structural mimicry of certain copper proteins (e.g. hemocyanin, tyrosinase and catechol oxidase), containing so-called type-3 copper binding sites, has been shown. This is a significant improvement since the use of scaffolded histidine residues is one step closer to the mimicry of enzymes by biological relevant species such as amino acids and peptides[
Exposure of DNA polymerase I to the protease subtilisin
Chemical Modification of the Fibrinolytic Serine Protease Subtilisin CIR and its Mutant Enzymes by Polyethylene Glycol.
Abstract; A potent fibrinolytic serine protease (Subtilisin CIR) of Bacillus subtilis CIR110 isolated from fermented food has been purified by hydrophobic column chromatography in high yields. The enzyme was proved to be a subtilisin-like serine protease with total 275 amino acid residues, containing 8 lysine residues, and it demonstrated high fibrinolytic activity irrespective of the presence of plasminogen. Furthermore we prepared a variety of Subtilisin CIR mutant enzymes having additional lysine residues, containing 9-11 lysine residues, introduced by using site-directed mutagenesis for polymer modification. For the application to antithrombogenic biomaterials, Subtilisin CIR and its mutants were modified by activated PEG2(2,4-bis(O-methoxypolyethylene glycol)-6-chloro-s-triazine, MW 5000*2), and both the native and PEG-modified enzymes were studied by activity assays. The PEG-modified enzymes showed high fibrinolytic activity, better storage stability, reduced antigenicity, and highly increased resistance to plasma protease inhibitors such as .ALPHA.2-macroglobulin, compared to unmodified enzymes. It is expected that the PEG-modified enzymes would find potential applications in antithrombogenic biomaterials.
Looking at this article, I see a similarity to what has already been created inside of Morgellons Disease... this is OLD news below. They are talking about using the same principles that are mostly likely closely involved in our disease:
First Live Organism with Synthetic Genome Created
20 May 2010
"Researchers then transplanted the artificially assembled genome into a M. capricolum cell that had been emptied of its own genome."
If we look at M. capricolum, we see this illustration which is, IMO - the same principle behind what we're seeing in the 'mite/louse' inside what I'm calling a baculoviral sphere:
"In contrast, the new study involves the transfer of pure DNA from simpler “prokaryotic” organisms, without any accompanying proteins or cellular structures. If the experiment failed, it would have suggested that DNA by itself is insufficient to take complete control over the cellular machinery of another cell.
The researchers transferred the genome of the goat pathogen Mycoplasma mycoides—genetically engineered to be antibiotic resistant—into the cell of a closely related species, M. capricolum.
Initially, the M. capricolum bacterium contained two genomes: its own and that of M. mycoides. But after the cell divided, one daughter cell inherited one genome and the other got the foreign genome. Both populations were then exposed to antibiotics, leaving only the hybrid bacteria alive.
The researchers are unsure whether their results can be replicated with eukaryotic cells.
Venter’s team aims to create a synthetic organism with the minimum set of genes necessary for life. This could help answer the question of what life is..."
This semen organism above (mite or louse-like) is coming from the sphere acting as a baculoviral capsid, this is not an external organism that happened to get caught up in the photographing. I have two other photographs that substantiate this observation that clearly show this - 'crab-like', 'lobster-like' organism coming out of the gel womb from inside what I previously called a sphere acting as a baculovirus, from a human arm lesion at 300x:
I wrote one of the prestigious professors at one of the major universities yesterday and asked them how can Venter be claiming to have newly invented this process, that this has already been done in science and sent them the photo above of the arm lesion (of course, he probably won't respond back).
We can also suspect from this fellow's 'louse/mite' in his semem that this 'technology' is at least 5 years old from his symptoms that he says came about immediately? That photograph that I put out of my friend's arm lesion is at least 1 year old. It makes one wonder if someone had prior knowledge of this technology or that what has happened to some of us - is a coincidential accident.
As you know, I am looking toward what is happening in nature for clues as to what has happened to us. We see that with the bees, their mites are out of order and killing them now. We are suspecting the miticide insecticides that have targeted the mites/lice in nature have thrown the balance of the entire 'food chain' out of whack. I am also suspecting that our own human demodex mites are somehow involved in our disease.
I have described our sphere/s as a 'Pac Man' dot eater that incorporates whatever dna that it comes into contact with - I can see this in its physical characteristics. These above 'mites' are now GM organisms and one person of knowledge said that they believed it is a pre-insect pupa, the only insect to have hatched from the arm lesion petri dish was a fungus gnat, no adult mites were seen.
I am seeing where one of the Borrelia pathogens can be transmitted by the louse:
"Borrelia is a genus of bacteria of the spirochete phylum. It causes borreliosis, a zoonotic, vector-borne disease transmitted primarily by ticks and some by lice, depending on the species.
Lice that feed on infected humans acquire the Borrelia organisms that then multiply in the gut of the louse. When an infected louse feeds on an uninfected human, the organism gains access when the victim crushes the louse or scratches the area where the louse is feeding. B. recurrentis infects the person via mucous membranes and then invades the bloodstream."
I'm thinking that both of these people in the above photographs with the 'mite/louse' most likely have had contact with the louse pathogenic form of Lyme Disease and should be tested and treated for this.
Hi, I am new here but not to Morg symptoms. Can you expand upon fungus gnats.? What do they look like? I am just about over my a second ground of infection and both times noticed a great many odd little flies around my house. At least I thought they were flies but they act differently. At times I think they are hatching out of my head!!!
Hi, I am new here but not to Morg symptoms. Can you expand upon fungus gnats.? What do they look like? I am just about over my a second ground of infection and both times noticed a great many odd little flies around my house. At least I thought they were flies but they act differently. At times I think they are hatching out of my head!!!
Any input would be appreciated, thanks Sharon
Hi Sharon! Try to relax with it, sweetie... I don't how to say this but to just lay it out here...
It seems that some of us have a transgenic 'organism' involved in our disease, some of us have a variety, we're not sure yet - we're still in discovery.
I have microscopically 'caught' the fungus gnat in action - appearing from what I call a baculoviral expression system coming out of the Morgellons crystals.
In what I observed, they originated from a crystal-shaped capsid (capsule)... I'm not sure what the 'critter' is coming from the spheres, to me - it looks like a mite or a louse.
I've tried to explain what I believe is happening in the Youtube movies - "The Life Cycle of Morgellons"... I think Aqt posted those on another thread or search Youtube under that name, there's a Part 1 and 2.
I believe the fungus gnat aspect has a yearly life cycle, that it appears as an adult in the end of July/August. Jo, who is the gnat/fly 'expert' of our group, tends to think that it can be seen earlier in the Spring, we're not sure yet - we're trying to reproduce experiments that show this - I'm waiting on 90+ degree temperatures and that time of year.
What do we know for sure - we have one Morgie that is producing live, adult insects from her sinuses, looks like it's in the beetle family, it is not a fungus gnat. I know of another who has them in her nose, which shows two different small flies - not necessarily fungus gnats. It has taken this sinus person many years to get to this stage of production, 10 years, I think. I'm not sure about the second, nose person - I need to ask them over how much time lapsed before they started seeing live insects?
For some - that if the life cycle is allowed to continue, without disturbing it with medications, herbs, bioset, rife, far infrared light etc., in a site in our bodies, we can produce an adult insect.
This sounds disturbing, and it is, however - to calm you - once again, these are transgenic genetically modified insects. They are not 'alive' until they are allowed to be. We have to believe that certain methods of treating ourselves prevents this aspect from happening. A lot of research has gone into how to deter them from growing, living, etc. which is posted on various sites and their threads, basically under 'how to threat lesions', 'what to put on the skin'...
Katinka and I kid each other about them hatching out of her head and out of my ear (I have a bad ear lesion which has produced live larvae in the past)... we are not just kidding but have to put it in a humorous way to cope, I hope you understand?
tash: Hi skizit, I have watched all your videos on youtube and cant thank you enough for all you have educated me on. I cry for you alot and a bit for me. I was wanting to send you photos of what is raining down everyday here in Australia in hope you can tell me
Dec 11, 2019 23:28:22 GMT -5
tash: not sure where to send them as hush mail and rocket mail bounced back
Dec 11, 2019 23:30:03 GMT -5