Role of Regulatory T Cells in Tubercular Uveitis

<p><i>Purpose</i>: To study the role of regulatory T cells (Tregs) in patients with tubercular uveitis.</p> <p><i>Methods</i>: Frequencies of peripheral Tregs, Th1, Th17 cells, and intracellular cytokines were determined in 17 tubercular uveitis patients and 18 disease controls. Function of Tregs, Th1, and Th17 cells was assessed <i>in vitro</i>. Simultaneously, ocular levels of IFN-γ, IL-17A, IL-4, and IL-10 were also measured.</p> <p><i>Results</i>: Frequencies of peripheral Tregs in tubercular uveitis subjects were significantly lower compared with disease controls. Furthermore, expression of TGF-β and IL-2Rα, but not CTLA4, was reduced in Tregs of the tubercular uveitis group. The tubercular uveitis group demonstrated heightened Th1, Th17 responses following <i>in vitro</i> stimulation with phorbol myristate acetate (PMA)/ionomycin. Interestingly, Treg suppression assay did not show a significant difference between the two groups. Ocular levels of IFN-γ, IL-17A, and IL-10 were also elevated in tubercular uveitis group.</p> <p><i>Conclusions</i>: Low Treg frequency and hyporesponsive function contribute to proinflammatory responses manifesting at ocular level in tubercular uveitis.</p

Assessment of Effluent Contaminants from Three Facilities Discharging Marcellus Shale Wastewater to Surface Waters in Pennsylvania

Unconventional natural gas development in Pennsylania has created a new wastewater stream. In an effort to stop the discharge of Marcellus Shale unconventional natural gas development wastewaters into surface waters, on May 19, 2011 the Pennsylvania Department of Environmental Protection (PADEP) requested drilling companies stop disposing their wastewater through wastewater treatment plants (WWTPs). This research includes a chemical analysis of effluents discharged from three WWTPs before and after the aforementioned request. The WWTPs sampled included two municipal, publicly owned treatment works and a commercially operated industrial wastewater treatment plant. Analyte concentrations were quanitified and then compared to water quality criteria, including U.S. Environmental Protection Agency MCLs and “human health criteria.” Certain analytes including barium, strontium, bromides, chlorides, total dissolved solids, and benzene were measured in the effluent at concentrations above criteria. Analyte concentrations measured in effluent samples before and after the PADEP’s request were compared for each facility. Analyte concentrations in the effluents decreased in the majority of samples after the PADEP’s request (<i>p</i> < .05). This research provides preliminary evidence that these and similar WWTPs may not be able to provide sufficient treatment for this wastewater stream, and more thorough monitoring is recommended

Kaplan-Meier survival curves.

<p>Data are presented for all genotypes for all mice (upper left), all mice segregated by sex (lower left), all male mice (upper right), and all female mice (lower right). In the lower left panel, female mice are designated by solid lines, and male mice by broken lines. <i>P</i><0.001 for all comparisons except <i>Tnnt2</i>^R141W/+ females versus wildtype females and wildtype males versus wildtype females. <i>n</i> = 84 wildtype males, 67 wildtype females, 79 <i>Tnnt2</i>^R141W/+ males, 100 <i>Tnnt2</i>^R141W/+ females.</p

Echocardiography at age 10 weeks in mice crossed with transgenic overexpressors of <i>Tnnt2</i> mRNA (TG^WT) or phospholamban knockout (<i>Pln</i>^-/-) mice.

<p>Echocardiography at age 10 weeks in mice crossed with transgenic overexpressors of <i>Tnnt2</i> mRNA (TG^WT) or phospholamban knockout (<i>Pln</i>^-/-) mice.</p

Representative APs recorded from wildtype and <i>Tnnt2</i>^R141W/+ hearts.

<p><b>(A)</b> Spontaneously beating <i>Tnnt2</i>^R141W/+ hearts had an intrinsically lower heart rate. <b>(B)</b> <i>Tnnt2</i>^R141W/+ hearts exhibited relatively small increases in heart rate at 1 and 10 μM concentrations.</p

Pacing kinetics.

<p>AP durations were shorter in <i>Tnnt2</i>^R141W/+ relative to wildtype hearts at all pacing cycle lengths. <i>n</i> = 4 for WT, <i>n</i> = 3 for all <i>Tnnt2</i>^R141W/+ because not all <i>Tnnt2</i>^R141W/+ hearts could be paced at faster rates or shorter cycle lengths. *, <i>P</i><0.001 vs. wildtype.</p

Normalized force-pCa relationships in wildtype and <i>Tnnt2</i>^R141W/+ skinned papillary muscle fibers.

<p>Normalized force (i.e., ratio of force at a given pCa and maximally activated force at pCa = 4.33) developed at a range of Ca²⁺ concentrations was assessed at sarcomere lengths 1.9 μm (A: wildtype ○, <i>Tnnt2</i>^R141W/+ ∇) and 2.3 μm (B: wildtype ▲, <i>Tnnt2</i>^R141W/+ ◆). There was a rightward shift of the force-pCa curve in <i>Tnnt2</i>^R141W/+ muscle, indicating Ca²⁺ desensitization. Values are mean±SE (<i>n</i> = 8 for wildtype and <i>n</i> = 3 for <i>Tnnt2</i>^R141W/+).</p

Representative Ca²⁺ transients from wildtype (WT) and <i>Tnnt2</i>^R141W/+ hearts.

<p><b>(A)</b> Langendorff perfused hearts were loaded with Rhod-2/AM and imaged on a 16x16 element photodiode array to map Ca²⁺ transients (CaT). Spontaneously beating, <i>Tnnt2</i>^R141W/+ hearts exhibited a markedly slower intrinsic sinus rhythm heart rate (71±12 bpm, top right traces) compared to wildtype hearts (343±52 bpm top left traces; <i>n</i> = 6 each, <i>P</i><0.03). Calibration of Rhod-2 through measurements of F_max (maximum Rhod-2 fluorescence when saturated with Ca²⁺) and F_min (minimum Rhod-2 fluorescence when all Ca²⁺ is washed out and chelated with EGTA) detected increases in diastolic and peak systolic cytosolic Ca²⁺ in <i>Tnnt2</i>^R141W/+ compared to wildtype hearts (lower left panel). <i>Tnnt2</i>^R141W/+ hearts had a longer rise time of [Ca²⁺]_i, (WT = 14.3±2.09 ms, <i>n</i> = 4 and <i>Tnnt2</i>^R141W/+ = 17.20±0.67, <i>n</i> = 6; <i>P</i><0.05), and a longer time to recover to diastolic[Ca²⁺]_i. Lower right traces show the superposition of normalized CaTs from a wildtype and a <i>Tnnt2</i>^R141W/+ heart recorded from the center of the left ventricles, when both hearts were paced at the same cycle length (350 ms). The superposition of CaTs exposes marked differences in Ca²⁺ dynamics associated with Ca²⁺ desensitization in <i>Tnnt2</i>^R141W/+ hearts. <b>(B)</b> <i>Tnnt2</i>^R141W/+ hearts exhibited lower intrinsic heart rates and lower peak heart rates in response to isoproterenol at 1 and 10 μM concentrations, compared to wildtype, <i>P</i><0.001.</p

Troponin I expression.

<p>Immunoblot and densitometry of phosphorylated troponin I (P-TnI) and total troponin I (TnI) in 8-week-old wildtype (WT) and <i>Tnnt2</i>^R141W/+ hearts. <i>n</i> = 3 for wildtype and <i>n</i> = 3 for <i>Tnnt2</i>^R141W/+.</p

Primer sequences used for real-time quantitative PCR (QPCR).

<p>Primer sequences used for real-time quantitative PCR (QPCR).</p