In vivo and in vitro evaluation of cell-mediated immunity in patients with paracoccidioidomycosis

The cellular immune response to specific and nonspecific agents was investigated. both in vivo and in vitro, in 19 patients with paracoceidioidomycosis. In addition, the immunologic study of an investigator aceidentally inoculated with P. brasiliensis was included in this study. Nearly half of the patients showed depressed cell-mediated immune responses, as evaluated by intradermal tests with an antigenic preparation from P. brasiliensis (P.b.Ag.), ubiquitous antigens, and by the ability to develop sensitization to 2,4-dinitrochlorobenzene. A similar proportion of impaired responses was observed when the patients' lymphocytes were cultured with phytohemagglutinin (PHA). C'. albicans antigen and P.h.Ag. A factor was detected in the plasma of some patients which reduced the ability of patients' and normal lymphocytes to undergo blastic transformation. A positive correlation was found between the ability to develop delayed cutaneous hypersensitivity reactions to P.b.Ag. and other ubiquitous antigens, normal in vitro responsiveness to PHA and the absence of humoral blastogenic inhibitory factor. The inhibition of leukocyte migration, but not lymphocyte transformation, correlated positively with delayed hypersensitivity. The percentage of T lymphocytes was significantly reduced in the group of patients, being the absolute number and percentage of B cells bearing receptors tor complement normal. Two polar immunological patterns emerged. One characterized by positiveness in the skin test to P.b.Ag. and lack of significant abnormalities in cellular immunity, and another anergic to P.b.Ag., with cell mediated immunity severely depressed. Between the two polar groups, there were patients with intermediary patterns of immune response. This paper also includes the results obtained with the administration of transfer factor and levamisole to some of the patients.     

Eveningness and poor sleep quality contribute to depressive residual symptoms and behavioral inhibition in patients with bipolar disorder

ABSTRACT

Eveningness and sleep disturbances are considered as markers of Bipolar Disorder (BD) and influence mood and emotional or behavioral states. This study investigates the associations between circadian markers and sleep quality on residual depressive symptoms and inhibition/activation dimensions during the euthymic phase. A sample of 89 euthymic adult individuals with BD was assessed for circadian preference and typology using the Composite Scale of Morningness (CSM) and the Circadian Type Inventory (CTI) and for sleep quality using the Pittsburgh Sleep Quality Index (PSQI). The Montgomery and Asberg Depression Rating Scale (MADRS) and the Multidimensional Assessment of Thymic States (MAThyS) were used to measure residual depressive symptoms and the inhibition/activation dimensions. We examined any associations between these parameters using correlations and path analyses. We identified significant associations between eveningness and poorer sleep quality that correlated to higher depressive residual symptoms and a global inhibition. The use of path analyses led us to conclude that poor sleep quality mediated the relationship between eveningness and either residual mood symptoms or behavioral inhibition (motivation, sensory perception, interpersonal interaction, and cognition). These factors should be considered in the clinical evaluation of individuals with BD, with a specific attention during the euthymic phase, in order to achieve the best functional outcome possible.     

Functional analyses of AGAMOUS family members in Nicotiana benthamiana clarify the evolution of early and late roles of C-function genes in eudicots

The C-function, according to the ABC model of floral organ identity, is required for stamen and carpel development and to provide floral meristem determinacy. Members of the AG lineage of the large MADS box gene family specify the C-function in a broadly conserved manner in angiosperms. In core eudicots, two sub-lineages co-exist, euAG and PLE, which have been extensively characterized in Antirrhinum majus and Arabidopsis thaliana, where strong sub-functionalization has led to highly divergent contributions of the respective paralogs to the C-function. Various scenarios have been proposed to reconstruct the evolutionary history of the euAG and PLE lineages in eudicots, but detailed functional analyses of the roles of these genes in additional representative species to validate evolutionary hypotheses are scarce. Here, we report functional characterization of euAG- and PLE-like genes in Nicotiana benthamiana through expression analyses and phenotypic characterization of the defects caused by their specific down-regulation. We show that both paralogs redundantly contribute to the C-function in this species, providing insights on the likely evolution of these gene lineages following divergence of the major groups within the eudicots (rosids and asterids). Moreover, we have demonstrated a conserved role for the PLE-like genes in controlling fruit dehiscence, which strongly supports the ancestral role of PLE-like genes in late fruit development and suggests a common evolutionary origin of late developmental processes in dry (dehiscent) and fleshy (ripening) fruits.     

Taking into account local practices and indigenous knowledge in an emergency conservation context in Madagascar

At the 5th World Parks Congress, held in Durban, South Africa in 2003, the President of Madagascar committed his government to tripling the country’s protected zones over the next 5 years. The announcement reflected a desire to combine rapid conservation efforts with sustainable development. Conservationists in Madagascar focused their attention on the endemic baobab tree, Adansonia grandidieri. This paper aims to identify the contradictions between the political emergency of the biodiversity conservation effort and local development needs. Eighty-three semi-structured interviews were conducted in two villages near the protected area of “Baobab Alley” in the Menabe region. Malagasy conservationists believed the area’s protected status would benefit the local economy through eco-tourism. However, the conservation actions undertaken there display limited understanding of local dynamics and conflict with farmers’ needs. To protect the baobabs, the government has prohibited rice cultivation without providing compensation. We show that the multifunctional baobab tree is integrated into an agroforestry system and protected by farmers. Based on these results, we address the issue of how to combine conservation and local development objectives through the involvement of farmers and the recognition of local knowledge in tree management. We also demonstrate that an emergency approach to conservation is not conducive to the successful integration of conservation and development.     

Gene electrotransfer: from biophysical mechanisms to in vivo applications

Gene electrotransfer can be obtained not just on single cells in diluted suspension. For more than 10 years, this is a quasi routine strategy in tissue on the living animal and a few clinical trials have now been approved. New problems have been brought by the close contacts of cells in tissue both on the local field distribution and on the access of DNA to target cells. They need to be solved to provide a further improvement in the efficacy and safety of protein expression. There is a competition between gene transfer and cell destruction. Nevertheless, present results are indicative that electrotransfer is a promising approach for gene therapy. High level and long-lived expression of proteins can be obtained in muscles. This is used for a successful method of electrovaccination.     

How β-Lactam Antibiotics Enter Bacteria: A Dialogue with the Porins

Background

Multi-drug resistant (MDR) infections have become a major concern in hospitals worldwide. This study investigates membrane translocation, which is the first step required for drug action on internal bacterial targets. β-lactams, a major antibiotic class, use porins to pass through the outer membrane barrier of Gram-negative bacteria. Clinical reports have linked the MDR phenotype to altered membrane permeability including porin modification and efflux pump expression.

Methodology/Principal Findings

Here influx of β-lactams through the major Enterobacter aerogenes porin Omp36 is characterized. Conductance measurements through a single Omp36 trimer reconstituted into a planar lipid bilayer allowed us to count the passage of single β-lactam molecules. Statistical analysis of each transport event yielded the kinetic parameters of antibiotic travel through Omp36 and distinguishable translocation properties of β-lactams were quantified for ertapenem and cefepime. Expression of Omp36 in an otherwise porin-null bacterial strain is shown to confer increases in the killing rate of these antibiotics and in the corresponding bacterial susceptibility.

Conclusions/Significance

We propose the idea of a molecular “passport” that allows rapid transport of substrates through porins. Deciphering antibiotic translocation provides new insights for the design of novel drugs that may be highly effective at passing through the porin constriction zone. Such data may hold the key for the next generation of antibiotics capable of rapid intracellular accumulation to circumvent the further development MDR infections.     

NRP/Optineurin Cooperates with TAX1BP1 to Potentiate the Activation of NF-κB by Human T-Lymphotropic Virus Type 1 Tax Protein

Nuclear factor (NF)-κB is a major survival pathway engaged by the Human T-Lymphotropic Virus type 1 (HTLV-1) Tax protein. Tax1 activation of NF-κB occurs predominantly in the cytoplasm, where Tax1 binds NF-κB Essential Modulator (NEMO/IKKγ) and triggers the activation of IκB kinases. Several independent studies have shown that Tax1-mediated NF-κB activation is dependent on Tax1 ubiquitination. Here, we identify by co-immunoprecipitation assays NEMO-Related Protein (NRP/Optineurin) as a binding partner for Tax1 in HTLV-1 infected and Tax1/NRP co-expressing cells. Immunofluorescence studies reveal that Tax1, NRP and NEMO colocalize in Golgi-associated structures. The interaction between Tax1 and NRP requires the ubiquitin-binding activity of NRP and the ubiquitination sites of Tax1. In addition, we observe that NRP increases the ubiquitination of Tax1 along with Tax1-dependent NF-κB signaling. Surprisingly, we find that in addition to Tax1, NRP interacts cooperatively with the Tax1 binding protein TAX1BP1, and that NRP and TAX1BP1 cooperate to modulate Tax1 ubiquitination and NF-κB activation. Our data strongly suggest for the first time that NRP is a critical adaptor that regulates the assembly of TAX1BP1 and post-translationally modified forms of Tax1, leading to sustained NF-κB activation.     

Consequence of omitting or adding a meal in man on body composition, food intake, and metabolism

Objective: To investigate in man the consequence on body composition and related biological and metabolic parameters of omitting or adding a meal. Research Methods and Procedures: Twenty‐four young normal‐weight male subjects were recruited, 12 usual four‐meal and 12 usual three‐meal eaters, differing only in the consumption of an afternoon meal. They omitted or added a fourth meal during a 28‐day habituation period and were asked to report their intake on three 3‐day occasions. Before and after this habituation period, subjects participated in a session with a time‐blinded procedure, and blood was collected continuously from lunch to the spontaneously requested dinner. Body composition, respiratory quotient, and biochemical parameters were measured in the late evening preceding each session. Results: Omitting a meal was followed by increases in fat mass (360 ± 115 grams, p < 0.05), late evening leptin concentration (20.7 ± 11.0%, p < 0.05), and respiratory quotient (3.7 ± 1.4%, p < 0.05). Increase in the percentage of dietary fat during the habituation period (+4.1 ± 2.0%, p < 0.05) was correlated with fat mass (r = 0.66, p < 0.05). Adding a meal had no effect, but, in both groups, the change in energy content at this fourth eating occasion was correlated with the change in adiposity. Discussion: Our results suggest that adiposity may increase when young lean male subjects switch from a four‐ to a three‐meal pattern by removing their usual afternoon meal. This effect could be partly mediated by a change in the macronutrient composition of the diet.     

Comparative water balance profiles of Orchidaceae seeds for epiphytic and terrestrial taxa endemic to North America

The Orchidaceae have dust-like seeds that use wind currents for long-distance dispersal. Lacking endosperm, orchid embryos consume free-living, mycorrhizal fungi as a carbon source (mycotrophy) after settling on a substrate. Few studies have investigated orchid seed morphology as it relates to ecology, but conceivably variations in seed size and testa characteristics could be linked to water loss rates aimed at maximizing germination in a particular habitat. Seeds of 2 epiphytic, 1 aquatic, and 7 terrestrial orchids native to North America were compared with respect to water balance profiles: Cleistes bifaria, Encyclia tampensis, Epidendrum nocturnum, Habenaria repens, Isotria medeoloides, Liparis elata, L. hawaiensis, Platanthera holochila, P. integrilabia, and P. leucophaea. Water content, water loss rate, activation energy, and equilibrium humidity were assessed for each species. Seeds of epiphytic orchids were smaller, lighter, more porous, and had higher water loss rates compared to terrestrials. No active mechanism for water absorption exists in seeds of either group. Water loss appears to be a species-specific phenomenon that may be linked to the ecological niches these species occupy.