Activation of Salmonella Typhi-Specific Regulatory T Cells in Typhoid Disease in a Wild-Type S. Typhi Challenge Model

Salmonella Typhi (S. Typhi), the causative agent of typhoid fever, causes significant morbidity and mortality worldwide. Currently available vaccines are moderately efficacious, and identification of immunological responses associated with protection or disease will facilitate the development of improved vaccines. We investigated S. Typhi-specific modulation of activation and homing potential of circulating regulatory T cells (T_reg) by flow and mass cytometry using specimens obtained from a human challenge study. Peripheral blood mononuclear cells were obtained from volunteers pre- and at multiple time-points post-challenge with wild-type S. Typhi. We identified differing patterns of S. Typhi-specific modulation of the homing potential of circulating T_reg between volunteers diagnosed with typhoid (TD) and those who were not (No TD). TD volunteers demonstrated up-regulation of the gut homing molecule integrin α4ß7 pre-challenge, followed by a significant down-regulation post-challenge consistent with T_reg homing to the gut. Additionally, S. Typhi-specific T_reg from TD volunteers exhibited up-regulation of activation molecules post-challenge (e.g., HLA-DR, LFA-1). We further demonstrate that depletion of T_reg results in increased S. Typhi-specific cytokine production by CD8+ T_EM in vitro. These results suggest that the tissue distribution of activated T_reg, their characteristics and activation status may play a pivotal role in typhoid fever, possibly through suppression of S. Typhi-specific effector T cell responses. These studies provide important novel insights into the regulation of immune responses that are likely to be critical in protection against typhoid and other enteric infectious diseases.     

Trade-off between plant growth and defense? A comparison of sagebrush populations

We used ecotypic variation in big sagebrush (Artemisia tridentata) to examine potential trade-offs between inherent growth rate and tolerance or resistance to herbivory. Seeds were obtained from seven geographic populations, and 1,120 seedlings were established in a common garden. In one set of plots, plants were subjected to five treatments: control, regular insecticide spray, moderate browsing, severe browsing, or moderate browsing plus insecticide. Plants in a second set of plots were all untreated, and were used to estimate ambient growth, flower production, and susceptibility to herbivorous insects. In the first growing season, population differences in relative growth rate produced approximately seven-fold variation in mean biomass. Two populations of basin big sagebrush (A. tridentata tridentata) and one population of mountain big sagebrush (A. tridentata vaseyana) grew fastest; those of Wyoming big sagebrush (A. tridentata wyomingensis) showed the slowest growth. Bi-weekly application of insecticide for two growing seasons had no effect on the growth of either browsed or unbrowsed plants. All populations showed compensatory growth (but not overcompensation) in response to browsing, but the degree of compensation was unrelated to inherent growth rate. Similarly, there was no consistent relationship between plant growth rate and flower production in the second growing season. Some insects colonized fast-growing populations more frequently than slow-growing ones, but patterns of insect colonization were species-specific. At the level of geographic populations and subspecies, we found little evidence of a built-in trade-off between inherent growth rate and the ability to tolerate or resist herbivory. Because population ranks for growth rate changed substantially between seasons, attempts to correlate growth and defense characters need to account for differences in the growth trajectories of perennial plants.     

Unique butyric acid incorporation patterns for salinosporamides A and B reveal distinct biosynthetic origins

Feeding sodium butyrate (0.25–1 mg/ml) to cultures of Salinispora tropica NPS21184 enhanced the production of salinosporamide B (NPI-0047) by 319% while inhibiting the production of salinosporamide A (NPI-0052) by 26%. Liquid chromatography mass spectrometry analysis of the crude extract from the strain NPS21184 fed with 0.5 mg/ml sodium [U-¹³C₄]butyrate indicated that butyrate was incorporated as a contiguous four-carbon unit into NPI-0047 but not into NPI-0052. Nuclear magnetic resonance analysis of NPI-0047 and NPI-0052 purified from the sodium [U-¹³C₄]butyrate-supplemented culture extract confirmed this incorporation pattern. The above finding is the first direct evidence to demonstrate that the biosynthesis of NPI-0047 is different from NPI-0052, and NPI-0047 is not a precursor of NPI-0052.     

Visual documentation of ovine pituitary gland development with magnetic resonance imaging following zeranol treatment

The objective of the current study was to determine whether magnetic resonance imaging (MRI) could be successfully utilized to document the effect of an oestrogenic anabolic agent on pituitary gland growth. The experimental animals consisted of two 1/2 sibling Suffolk wethers (castrated rams), which received either no implant (control, n = 1) or a 24 mg zeranol implant at day 0 and day 42 (zeranol; n = 1). Animals were anaesthetized with propofol and supported with oxygen during the MRI procedure. A mobile MRI unit with a 0.5 tesla (T), superconducting magnet was used to obtain 3 mm thick, non-contrast enhanced, T1-weighted (TR 500-600, TE25) sagittal, transverse and dorsal images of the pituitary gland. Sagittal images were recorded only when the mesencephalic aqueduct and infundibulum were distinctly visible in the same image. Pituitary glands were imaged at 14-day intervals for 70 days to determine if and when the anabolic effects of zeranol on pituitary gland growth could be visualized using MRI techniques. Three separate measurements of the pituitary gland dimensions made with the on-screen cursor were averaged to calculate pituitary gland dimensions and volume. A computer-assisted image analysis system and laser film images were used to determine pituitary gland area. Increases in pituitary gland volume for control and zeranol-treated animals were evident within 14 days, and by the end of the 70-day study, the increase in pituitary volume for the zeranol-treated animal was three times greater than that of the control animal. Overall, our results indicate that MRI technology can be successfully used to document the development of the pituitary gland in vivo. Application of knowledge gained from this novel approach to study the growth, development and function of endocrine glands over time, and within the same animal, will enhance human and animal endocrine diagnostic procedures.     

Contributions of Fe(III) to UV–Vis absorbance in river water: a case study on the Connecticut River and argument for the systematic tandem measurement of Fe(III) and CDOM

Biogeochemistry - Dissolved organic matter (DOM) impacts the structure and function of aquatic ecosystems. DOM absorbs light in the UV and visible (UV–Vis) wavelengths, thus impacting light...     

Differential distribution of leaf chemistry in eucalypt seedlings due to variation in whole-plant nutrient availability

We investigated the effects of whole-plant nutrient-availability on the degree of distribution of some plant primary and secondary (nitrogen, fibre, flavonols, gallotannins and cineole) chemicals across young, mature and old leaves of seedlings of Eucalyptus nitens. Four treatments that ranged from low to high nutrient-application rates resulted in mean whole-plant foliar concentrations of 0.63%, 0.85%, 1.11% and 1.82% nitrogen dry matter (N%DM) for treatments A, B, C and D, respectively. Within-plant distribution (across the leaf age profile of young, mature and old leaves within a eucalypt seedling) of N%DM ranged from zero in treatment A to a wide range of distribution in treatment D (low N%DM concentrations in old leaves to high N%DM concentrations in young leaves). Similarly, the distribution of fibre ranged from zero in treatment A to a wide range of distribution in treatment D, but with high concentrations in old leaves and low concentrations in young leaves. In contrast, flavonols (weakly) and gallotannins had a wide range of distribution in treatment A (low concentrations in old leaves to high concentrations in young leaves) but were little or not distributed in the other treatments. Again in contrast, cineole was strongly distributed between old and young leaves (low concentrations in old leaves to high concentrations in young leaves) across all treatments while concentrations in mature leaves reflected one or other leaf age depending on treatment. Protein precipitable phenols in treatments A, B and C were high in young, and low in old leaves; whereas in treatment D they were low and similar between leaves of different ages.     

Novel racemosin B derivatives as new therapeutic agents for aggressive breast cancer

Carbazole derivatives show anti-cancer activity and are of great interest for drug development. In this study, we synthesized and analyzed several new alkylamide derivatives of racemocin B, a natural indolo[3,2-a]carbazole molecule originally isolated from the green alga Caulerpa racemose. Several alkylamide derivatives were found to exhibit moderate to strong growth inhibition against human breast cancer cell lines. They induced G2/M cell cycle arrest and apoptosis in the aggressive triple-negative breast cancer cell line MDA-MB-231. Among these derivatives, compound 25 with the lowest IC₅₀ induced cell death by suppressing autophagy. This was accompanied by inhibition of autophagic flux and accumulation of autophagy protein 1 light chain 3, LC3II, and p62. The novel alkylamide derivative offers a potential new treatment for human breast cancer.     

CDC25B Overexpression Stabilises Centrin 2 and Promotes the Formation of Excess Centriolar Foci

CDK-cyclin complexes regulate centriole duplication and microtubule nucleation at specific cell cycle stages, although their exact roles in these processes remain unclear. As the activities of CDK-cyclins are themselves positively regulated by CDC25 phosphatases, we investigated the role of centrosomal CDC25B during interphase. We report that overexpression of CDC25B, as is commonly found in human cancer, results in a significant increase in centrin 2 at the centrosomes of interphase cells. Conversely, CDC25B depletion causes a loss of centrin 2 from the centrosome, which can be rescued by treatment with the proteasome inhibitor MG132. CDC25B overexpression also promotes the formation of excess centrin 2 “foci”. These foci can accumulate other centrosome proteins, including γ-tubulin and PCM-1, and can function as microtubule organising centres, indicating that these represent functional centrosomes. Formation of centrin 2 foci can be blocked by specific inhibition of CDK2 but not CDK1. CDK2-mediated phosphorylation of Monopolar spindle 1 (Mps1) at the G1/S transition is essential for the initiation of centrosome duplication, and Mps1 is reported to phosphorylate centrin 2. Overexpression of wild-type or non-degradable Mps1 exacerbated the formation of excess centrin 2 foci induced by CDC25B overexpression, while kinase-dead Mps1 has a protective effect. Together, our data suggest that CDC25B, through activation of a centrosomal pool of CDK2, stabilises the local pool of Mps1 which in turn regulates the level of centrin 2 at the centrosome. Overexpression of CDC25B may therefore contribute to tumourigenesis by perturbing the natural turnover of centrosome proteins such as Mps1 and centrin 2, thus resulting in the de novo assembly of extra-numerary centrosomes and potentiating chromosome instability.