Deubiquitylating enzyme, USP9X, regulates proliferation of cells of head and neck cancer lines.

OBJECTIVES: Truncating mutations in USP9X have been identified in oral squamous cell carcinoma patients. The aim of this study was to determine USP9X's functional role, if any, in head and neck cancer cells. MATERIALS AND METHODS: USP9X was depleted/overexpressed in head and neck cancer cell line: SCC15 (tongue), CAL27 (tongue), FaDu (pharynx) and Detroit 562 (pharynx). Cell proliferation was monitored using the CyQUANT assay, and cell cycle distribution was determined by flow cytometry. Immunoblot assays were conducted to assess protein levels. RT-qPCR was performed to determine Notch and Wnt pathway target gene expression. RESULTS: Our data showed a direct correlation between USP9X protein levels and proliferation, as well as Notch pathway activity in head and neck cancer cells. However, at least in FaDu, USP9X did not appear to regulate proliferation through the Notch pathway. Immunoblotting revealed a dramatic reduction in downstream targets of mTOR complex 1, namely total ribosomal protein (S6) and its phosphorylated form (pS6), when USP9X was depleted in FaDu cells. In contrast, in immortalized but non-tumorigenic HaCaT keratinocytes, USP9X depletion led to increase in cell proliferation, maintaining direct regulation of Notch activity. CONCLUSIONS: The functional role of USP9X was found to be context dependent. USP9X possibly promotes head and neck cancer cell proliferation through the mTOR pathway.

Rickettsioses in the central hills of Sri Lanka: serological evidence of increasing burden of spotted fever group.

OBJECTIVES: To study the epidemiology, clinical features, and changing pattern of rickettsial infections on the western slopes of the hilly Central Province of Sri Lanka over 6 years. METHODS: All patients with rickettsial infections who presented to the Teaching Hospital, Peradeniya were studied prospectively from January 2002 to December 2007. An immunofluorescent antibody assay (IFA) was used to confirm the diagnosis. RESULTS: Of the 371 clinical cases of rickettsial infection, 122 underwent IFA to confirm the diagnosis. Species-specific IgG antibodies were positive in 105/122 (86.1%) cases: 43/105 (40.9%) to Rickettsia conorii and 6/105 (5.7%) to Orientia tsutsugamushi, with mixed antibody reactivity to more than one antigen in 56/105 (53.3%) cases, including Rickettsia typhi in 27/105 (25.7%). Among those with mixed IgG reactivity, IgM assays were used to detect pathogens responsible for acute infections. Finally, a total of 55 spotted fever group (SFG) infections, seven scrub typhus infections, and one case of murine typhus were identified. Of the 105 positive cases, 53 (50.5%) were male and 52 (49.5%) were female, and the mean age was 40 years (range 11-83 years). In the SFG patients, 13/55 (24%) had severe vasculitis with fern leaf type skin necrosis and 17/55 (31%) had arthritis. Three patients (5%) had an altered level of consciousness. A patient with scrub typhus had transient deafness. None of the 105 patients had an eschar. CONCLUSIONS: It appears that SFG rickettsioses are on the rise in the hilly Central Province of Sri Lanka, whilst murine typhus and scrub typhus are present at a lower rate.

Evidence of acute rickettsioses among patients presumed to have chikungunya fever during the chikungunya outbreak in Sri Lanka.

BACKGROUND: Chikungunya fever (CGF) and rickettsioses are known to cause acute onset febrile illnesses associated with severe arthritis. Rickettsial arthritis is curable with the use of appropriate anti-rickettsial antibiotics, however the arthritis of CGF tends to have a prolonged course leading to protracted disability. The aim of this study was to investigate the contribution of CGF and rickettsioses to cases of fever and arthritis during a presumed CGF outbreak in Sri Lanka. METHODS: Fifty-eight consecutive patients with presumed CGF were further investigated to determine the occurrence of rickettsioses among them, and to identify differences in clinical, hematological, and biochemical parameters between the two diseases. RESULTS: Nearly a third of the patients had serological evidence of rickettsioses accounting for their illness. The presence of a late onset major joint arthropathy sparing the small joints of the hands and feet, and the occurrence of a late onset discrete maculopapular rash over the trunk and extremities, suggested rickettsioses over CGF. White blood cell count, erythrocyte sedimentation rate, C-reactive protein, and liver function tests were not helpful in differentiating rickettsioses from CGF. Patients with rickettsioses and arthritis who received an empirical course of doxycycline recovered faster than those who did not receive specific treatment. CONCLUSIONS: The establishment of rapid diagnostic methods able to differentiate the etiological agents of fever and arthritis, such as CGF and rickettsioses, would be beneficial in endemic settings.

Contribution of rickettsioses in Sri Lankan patients with fever who responded to empirical doxycycline treatment.

Twenty-eight febrile Sri Lankan patients with undiagnosed fever for 7 days after hospital admission, who responded to empirical treatment with doxycycline, were retrospectively investigated using microimmunofluorescence assay to verify whether they had rickettsial infection. Eleven (39%) patients were confirmed as having spotted fever group rickettsioses and 10 (36%) as having Orientia tsutsugamushi. Seven were negative for all tests. This suggests that greater use of doxycycline appears justified for patients with undiagnosed fever in settings where rickettsial diseases are endemic or re-emerging with inadequate diagnostic facilities.