NOX4-driven ROS formation mediates PTP inactivation and cell transformation in FLT3ITD-positive AML cells.

Activating mutations of FMS-like tyrosine kinase 3 (FLT3), notably internal tandem duplications (ITDs), are associated with a grave prognosis in acute myeloid leukemia (AML). Transforming FLT3ITD signal transduction causes formation of reactive oxygen species (ROS) and inactivation of the protein-tyrosine phosphatase (PTP) DEP-1/PTPRJ, a negative regulator of FLT3 signaling. Here we addressed the underlying mechanisms and biological consequences. NADPH oxidase 4 (NOX4) messenger RNA and protein expression was found to be elevated in FLT3ITD-positive cells and to depend on FLT3ITD signaling and STAT5-mediated activation of the NOX4 promoter. NOX4 knockdown reduced ROS levels, restored DEP-1 PTP activity and attenuated FLT3ITD-driven transformation. Moreover, Nox4 knockout (Nox4(-/-)) murine hematopoietic progenitor cells were refractory to FLT3ITD-mediated transformation in vitro. Development of a myeloproliferative-like disease (MPD) caused by FLT3ITD-transformed 32D cells in C3H/HeJ mice, and of a leukemia-like disease in mice transplanted with MLL-AF9/ FLT3ITD-transformed murine hematopoietic stem cells were strongly attenuated by NOX4 downregulation. NOX4-targeting compounds were found to counteract proliferation of FLT3ITD-positive AML blasts and MPD development in mice. These findings reveal a previously unrecognized mechanism of oncoprotein-driven PTP oxidation, and suggest that interference with FLT3ITD-STAT5-NOX4-mediated overproduction of ROS and PTP inactivation may have therapeutic potential in a subset of AML.

Non-infected carotid artery pseudoaneurysm 29 years after endarterectomy, endovascular management with covered stent.

Pseudoaneurysm formation is a rare complication following carotid endarterectomy (CEA). Arterial pseudoaneurysms lack all three layers of the arterial wall that include the intima, media and adventitia. Pseudoaneurysms are most commonly seen after injuries to the artery in the form of blunt trauma and puncture, and are less common after surgeries such as carotid endarterectomy. These lesions present most frequently as enlarging, pulsatile, expandable masses associated with swelling and pain. Management of this complication is challenging. Traditionally, open surgical repair has been the preferred treatment. Recently, endovascular techniques using stent graft implantation alone or combined graft and coil embolization have offered a less invasive approach for the management of this lesion. Pseudoaneurysm development has been described within hours to several years after initial arterial injury, normally presenting within 5 years. To our knowledge, this is the first case report of pseudoaneurysm formation in a patient presenting 29 years after a carotid endarterectomy; during that time the patient remained completely asymptomatic until 2 months prior to his admission. The patient is an 84-year-old male with a history of stroke which prompted a left carotid endarterectomy in 1981. Twenty-nine years post procedure it was noted that the patient had a lump that was progressively enlarging on the left side of his neck, zone 1. It was pulsatile on examination. MRI/A imaging suggested a left carotid bulb aneurysm. The consulting vascular surgeon felt the patient would not be a good surgical candidate and so stenting was considered. Carotid and cerebral angiogram demonstrated a large 6 cm left carotid pseudoaneurysm off the carotid bulb. The diagnostic procedure was followed by a successful placement of an 8 x 10 cm Viabahn covered stent from the left common carotid artery to the left internal carotid artery. Following the procedure, the carotid artery was patent and there was minimal to no further residual filling of the pseudoaneurysm. Poststenting, the patient remained at his neurological baseline. This case demonstrates that pseudoaneurysm formation can occur as a long term complication after carotid endarterectomy. It may present as a rapidly expandable, pulsatile, vascular lesion in the absence of clinical and sub-clinical infection. Placement of an endovascular stent graft may be a safe and effective option for treatment of infected and non-infected carotid pseudoaneurysm.

Transarterial embolization of a cervical dural arteriovenous fistula. Presenting with subarachnoid hemorrhage.

SUMMARY: We describe a case of a 75-year-old man who presented with acute onset of headache and subarachnoid hemorrhage and initial cerebral angiography was deemed "negative". In retrospect, a faint contrast collection was present adjacent to the right vertebral artery at the C1 level suspicious for a small dural arteriovenous fistula (dAVF). Follow-up angiography with selective microcatheter injections of the right vertebral artery and C1 radicular artery confirmed a complex dAVF with characteristically specific venous drainage patterns associated with a subarachnoid hemorrhage presentation. Subsequently, the cervical dAVF was treated with superselective glue embolization resulting in complete occlusion. Cervical dAVFs are extremely rare vascular causes of subarachnoid hemorrhage. Both diagnostic angiography and endovascular treatment of these lesions can be challenging, especially in an emergent setting, requiring selective evaluation of bilateral vertebral arteries and careful attention to their cervical segments. Although only a single prior case of a cervical dAVF presenting with subarachnoid hemorrhage has been successfully treated with embolization, modern selective transarterial techniques may allow easier detection and treatment of subtle pathologic arteriovenous connections.

Physical mapping of the serotonin 5-HT(7) receptor gene (HTR7) to chromosome 10 and pseudogene (HTR7P) to chromosome 12, and testing of linkage disequilibrium between HTR7 and autistic disorder.

The gene encoding the serotonin 5-HT(7) receptor (HTR7) has been considered as a candidate locus in several neuropsychiatric disorders, based on pharmacological evidence and ligand-binding studies. After determining over 3 kb of previously unpublished sequence from introns 1 and 2 of HTR7, a single base (C/T) polymorphism in the second intron of HTR7 was found. Allele-specific PCR was used to genotype the HTR7 marker in 53 trios consisting of subjects with autistic disorder and both parents. Using the transmission disequilibrium test (TDT), no evidence of preferential transmission of either allele was found (TDT chi(2) = 0.252, p = 0.602). Sequence data obtained from both intron 1 and intron 2 of HTR7, and from the 5-HT(7) pseudogene (HTR7P), was used to confirm localization of HTR7 to 10q23 and HTR7P to 12p13 using radiation hybrid analyses.

Inhibition of cellobiohydrolase I from Trichoderma reesei by palladium.

Cellulase from Trichoderma reesei is a multienzyme mixture that hydrolyzes cellulose to glucose. Two enzymes in this mixture, cellobiohydrolase (CBH) and endoglucanase (EG), possess a common structure comprising a distinct cellulose-binding domain (CBD) and catalytic domain. Inhibition of the catalytic domain of cellulases without affecting their CBD function might be useful for structure/function studies of these enzymes. Complexes of the platinum group metals were tested for their ability to inhibit the major cellulase enzyme from T. reesei, cellobiohydrolase I (CBH I). Only palladium complexes inhibited CBH I, inhibition being dependent upon the molar ratio of palladium to CBH I with 1 microM CBH I retaining only 10% of its activity in the presence of 100 microM ammonium hexachloropalladate(IV) and after the incorporation of 28 mol Pd/mol CBH I. Inhibition was irreversible and could be completely prevented by including histidine, cysteine, and cystine in the assay mixture. Although the primary mechanism of inhibition of CBH I by palladium remains to be elucidated, it could involve the binding of palladium to sulfur or cystine residues resulting in their degradation. This is based on the findings that (i) palladium-inhibited CBH I was less thermally stable than native CBH I; (ii) CBH I, chemically modified by the attachment of pentaammine ruthenium(III) to the imidazole-N of either H206 or H228, showed greater sensitivity to inhibition by palladium compared to native CBH I; and (iii) ammonium hexachloropalladate cleaved 5,5'-dithiobis(2-nitrobenzoic acid)--Ellman's reagent. Binding of CBH I to crystalline cotton linters was not affected by palladium.

Palladium--a new inhibitor of cellulase activities.

Palladium complexes have been shown to strongly inhibit cellobiohydrolase I (CBH I) and endoglucanase II (EG II), two cellulases produced by Trichoderma reesei. Also inhibited were total cellulase (Avicelase) and beta-glucosidase (cellobiase) activities. The catalytic domain of CBH II, the second most abundant component of this cellulase, appeared less susceptible to inhibition by palladium. The inhibition was irreversible and could be prevented if histidine, cysteine or cystine was added to the enzyme reaction mixture simultaneously with the inhibitor. The binding of CBH I to microcrystalline cellulose (Avicel) was unaffected by palladium.