Surface colonization by Azospirillum brasilense SM in the indole-3-acetic acid dependent growth improvement of sorghum

The key to improving plant productivity is successful bacterial-plant interaction in the rhizosphere that can be maintained in the environment. The results presented here confirm Azospirillum brasilense strain SM as a competent plant growth promoting bacterium over mid- and long-term associations with sorghum. This study establishes that plant growth can be directly correlated with the associated bacterium's indole-3-acetic acid (IAA) production capability as IAA over-expressing variants, SMp30 and SMΔi3-6 fared better than the wild type strain. The auxin antagonist, p-chlorophenoxy isobutyric acid confirmed the role of bacterial IAA in plant growth promotion and verified the presence of larger amount of IAA available to the seeds on inoculation with IAA over-expressing mutants. Microscopic analysis identified the bacterial association at root tips, root-shoot junction and elongation zone and their surface colonizing nature. Scanning electron microscopy identified larger number of root hairs and extensive exopolysaccharide covering in comparison to untreated ones. In addition, vibroid-shaped Azospirilla attached by means of fibrillar material were dispersed along the elongation zone. The notable difference with IAA over-expressing variants was enhanced number of root hairs. Thus, the variant strains may be more efficient surface colonizers of the sorghum root and used as superior bio-inoculants for improving plant productivity.     

Breast milk as a source for acquisition of cytomegalovirus (HCMV) in a premature infant with sepsis syndrome: detection by real-time PCR.

Symptomatic disease due to human cytomegalovirus (HCMV) has been increasingly recognized in low birth weight (LBW) premature infants. Breast milk has been identified as a potential source for these infections. At approximately 60 days of life a LBW, premature infant was diagnosed with HCMV sepsis syndrome, prompting further investigation of the source for HCMV acquisition. The infant had received a combination of both fresh and frozen breast milk from his mother throughout his first 2 months of life. We utilized a real-time PCR assay for HCMV DNA to retrospectively examine viral genome copy number in serial samples of stored, frozen breast milk. This analysis indicated an increase in breast milk viral load over the first 45 days of life, heralding the HCMV sepsis syndrome that was observed clinically. Real-time PCR may be a useful tool in the evaluation of HCMV viral load in breast milk.     

Nonporous polyurethane membranes as islet immunoisolation matrices--biocompatibility studies

Novel elastomeric nonporous polyurethane membranes were synthesised with differing hard segment contents for evaluation as possible islet encapsulation matrices. Physico-chemical properties of these membranes were reported earlier by authors and have been found suitable for immunoisolation. In the present study, membranes were evaluated for their in vitro biocompatibility. Membranes T1, T4, T5 and T6 did not show toxicity in direct cell contact study towards L929 fibroblasts. However, T2 and T3 were found cytotoxic and were excluded from further testing. NIH3T3 cells when exposed to leach out products of T4, T5 and T6 showed no cytotoxicity, while T1 decreased cellular viability as confirmed by MTT assay. T4 and T5 alone were seen to be compatible with mouse islets while T6 was incompatible to the mouse islets. Digital image analysis (DIA) studies showed intact morphology of islets cultured on the T4 and T5 with viability (88.4 and 91% respectively) comparable to islets on tissue culture polystyrene (TCPS) control. Islets on T4 and T5 also retained their functionality, as judged by insulin secretion in response to in vitro glucose challenge (16.0 mM). These studies point out the crucial role of surface free energy and hydrophilicity in deciding compatibility of polyurethane membranes with islets of Langerhans. Studies indicate that polyurethane membranes T4 and T5 could be potential candidates for islet immunoisolation.     

“Breastfeeding” without baby: A longitudinal,qualitative investigation of how mothers perceive,feel about,and practice human milk expression

Most American mothers who produce human milk (HM) now pump in place of some or all feeding at the breast, and most American infants are now fed pumped HM. We aimed to investigate mothers' perceptions of, attitudes toward, and practices for pumping and providing pumped HM. Results related to pumping are reported here. We conducted in‐depth, semi‐structured interviews among a diverse sample of 20 mothers who pumped, following each from pregnancy through infant HM‐feeding cessation up to 1 year postpartum. Data were analyzed using thematic analysis with Atlas.ti. Mothers' reasons for pumping changed over time and reflected their needs and desires (e.g., latch difficulty, return to work, and increasing their milk supply). Mothers reported that pump type and quality were important to pumping success and that pumping was time‐consuming, costly, and unpleasant compared to feeding at the breast. Regardless of how often mothers pumped, most felt pumping was necessary to meet their infant HM‐feeding goals and was a welcome means of sharing with other caregivers the bonding opportunity and tasks they associated with feeding infants. Mothers interpreted output from pumping sessions to understand their ability to provide enough milk to meet their infants' needs. Mothers' reasons for pumping may signal constraints to infant HM feeding that may be addressed with policy changes. Mothers' attitudes and perceptions toward pumping indicate that, although pumping fills important and welcome roles for many mothers, the reality of its practice may make it an unacceptable or infeasible substitute for some.     

Differences in both inositol 1,4,5-trisphosphate mass and inositol 1,4,5-trisphosphate receptors between normal and dystrophic skeletal muscle cell lines

Human normal (RCMH) and Duchenne muscular dystrophy (RCDMD) cell lines, as well as newly developed normal and dystrophic murine cell lines, were used for the study of both changes in inositol 1,4,5-trisphosphate (IP₃) mass and IP₃ binding to receptors. Basal levels of IP₃ were increased two- to threefold in dystrophic human and murine cell lines compared to normal cell lines. Potassium depolarization induced a time-dependent IP₃ rise in normal human cells and cells of the myogenic mouse cell line (129CB3), which returned to their basal levels after 60 s. However, in the human dystrophic cell line (RCDMD), IP₃ levels remained high up to 200 s after potassium depolarization. Expression of IP₃ receptors was studied measuring specific binding of ³H-IP₃ in the murine cell lines (normal 129CB3 and dystrophic mdx XLT 4-2). All the cell lines bind ³H-IP₃ with relatively high affinity (K_d: between 40 and 100 nmol/L). IP₃ receptors are concentrated in the nuclear fraction, and their density is significantly higher in dystrophic cells compared to normal. These findings together with high basal levels of IP₃ mass suggest a possible role for this system in the deficiency of intracellular calcium regulation in Duchenne muscular dystrophy. © 1998 John Wiley & Sons, Inc. Muscle Nerve 21:902–909, 1998.     

Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

Plasmacytoid dendritic cells (PDC) are potent producers of alpha interferon (IFN-α) in response to enveloped viruses and provide a critical link between the innate and adaptive immune responses. Although the loss of peripheral blood PDC function and numbers has been linked to human immunodeficiency virus (HIV) progression in humans, a suitable animal model is needed to study the effects of immunodeficiency virus infection on PDC function. The rhesus macaque SIV model closely mimics human HIV infection, and recent studies have identified macaque PDC, potentially making the macaque a good model to study PDC regulation. In this study, we demonstrate that peripheral blood PDC from healthy macaques are both phenotypically and functionally similar to human PDC and that reagents used for human studies can be used to study macaque PDC. Both human and macaque PBMC expressed IFN-α in response to herpes simplex virus (HSV), the prototypical activator of PDC, as measured by using an IFN bioassay and IFN-α-specific enzyme-linked immunospot assays. Similar to human PDC, macaque PDC were identified by using flow cytometry as CD123⁺ HLA-DR⁺ lineage⁻ cells. In addition, like human PDC, macaque PDC expressed intracellular IFN-α, tumor necrosis factor alpha, macrophage inflammatory protein 1β/CCL4, and IFN-inducible protein 10/CXCL10 upon stimulation with HSV, all as determined by intracellular flow cytometry. We found that IFN regulatory factor 7, which is required for the expression of IFN-α genes, was, similar to human PDC, expressed at high levels in macaque PDC compared to monocytes and CD8⁺ T cells. These findings establish the phenotypic and functional similarity of human and macaque PDC and confirm the utility of tools developed for studying human PDC in this animal model.