Evaluation of Integrated Community Case Management in Eight Districts of Central Uganda

Objective

Evidence is limited on whether Integrated Community Case Management (iCCM) improves treatment coverage of the top causes of childhood mortality (acute respiratory illnesses (ARI), diarrhoea and malaria). The coverage impact of iCCM in Central Uganda was evaluated.

Methods

Between July 2010 and December 2012 a pre-post quasi-experimental study in eight districts with iCCM was conducted; 3 districts without iCCM served as controls. A two-stage household cluster survey at baseline (n = 1036 and 1042) and end line (n = 3890 and 3844) was done in the intervention and comparison groups respectively. Changes in treatment coverage and timeliness were assessed using difference in differences analysis (DID). Mortality impact was modelled using the Lives Saved Tool.

Findings

5,586 Village Health Team members delivered 1,907,746 treatments to children under age five. Use of oral rehydration solution (ORS) and zinc treatment of diarrhoea increased in the intervention area, while there was a decrease in the comparison area (DID = 22.9, p = 0.001). Due to national stock-outs of amoxicillin, there was a decrease in antibiotic treatment for ARI in both areas; however, the decrease was significantly greater in the comparison area (DID = 5.18; p<0.001). There was a greater increase in Artemisinin Combination Therapy treatment for fever in the intervention areas than in the comparison area but this was not significant (DID = 1.57, p = 0.105). In the intervention area, timeliness of treatments for fever and ARI increased significantly higher in the intervention area than in the comparison area (DID = 2.12, p = 0.029 and 7.95, p<0.001, respectively). An estimated 106 lives were saved in the intervention area while 611 lives were lost in the comparison area.

Conclusion

iCCM significantly increased treatment coverage for diarrhoea and fever, mitigated the effect of national stock outs of amoxicillin on ARI treatment, improved timeliness of treatments for fever and ARI and saved lives.     

Transient gene expression mediated by integrase-defective retroviral vectors

Nonintegrating retroviral vectors were produced from a Moloney murine leukemia virus (MoMLV)-based retroviral vector system by introducing a point mutation into the integrase (IN) gene of the packaging plasmid. The efficacy of IN-defective retroviral vectors was measured through the transient expression of ZsGreen or luciferase in human cell lines. The IN-defective retroviral vectors could transduce target cells efficiently, but their gene expression was transient and lower than that seen with the integrating vectors. IN-defective retroviral vector gene expression decreased to background levels in fewer than 10 days. Southern blot analysis of transduced K562 cells confirmed the loss of a detectable vector sequence by 15 days. The residual integration activity of the IN-defective vector was 1000- to 10,000-fold lower than that of the integrating vector. These results demonstrate that the IN-defective retroviral vectors can provide a useful tool for efficient transient gene expression targeting of primary hematopoietic stem cells and lymphoid cells.     

Flavonoid compounds related to seed coat color of wheat

In red wheat, reddish-brown pigments accumulate in testa of mature seeds. Half-cut wheat seeds were immersed in p-dimethylaminocinnamaldehyde (DMACA) reagent that stains flavanol structures blue. Testa of 10–40 days after flowering (DAF) in red wheat (“Norin 61” and “Satonosora”) seeds were stained blue and the reagent color changed to blue with 10–25 DAF seeds. No blue staining was observed in white wheat (“Tamaizumi”) seeds during maturation. “Norin 61” seed coats at 10 DAF contained dihydroquercetin, dihydromyricetin, (+)-catechin, procyanidin B3, and prodelphinidin B3, which were identified by HPLC-diode array detector and LC-MS/MS analyses. These five components began accumulating 7 DAF, reached maxima at 10 or 15 DAF, and then decreased in red wheat seeds, but were not detected in white wheat seeds. These results suggest that flavanol and proanthocyanidins are possible precursors of the reddish-brown pigments of red wheat seeds, and are converted to insoluble compounds as the seeds mature.     

Mapping QTLs for grain dormancy on wheat chromosome 3A and the group 4 chromosomes,and their combined effect

A major QTL for grain dormancy, QPhs.ocs-3A.1, derived from the highly dormant wheat Zenkoujikomugi (Zen), has been identified in a study made under a controlled environment. Further investigations were needed to dissect the precise position and expression of QPhs.ocs-3A.1 under different field conditions because the ability to detect genetic loci for grain dormancy traits is compromised by environmental effects and genotype/environment interactions. Group 4 chromosomes have also been shown to be possible sites of QTLs for grain dormancy. The objectives of this study were (1) to locate additional molecular markers in the QPhs.ocs-3A.1 region, (2) to identify QTLs on the group 4 chromosomes and (3) to elucidate their combined effects. We examined the recombinant inbred lines (RILs) from a cross between Chinese Spring (CS) and Zen over a 3-year period in one location and 1 year in a different location. In an interval mapping study QPhs.ocs-3A.1 was mapped to within the 4.6 cM region flanked by Xbarc310 and Xbcd907 at the proximal end of the short arm of chromosome 3A. QPhs.ocs-3A.1 was confirmed to be the predominant dormancy QTL since it explained a large portion (11.6–44.8%) of the phenotypic variation, and was strongly displayed under dormancy-breaking conditions or at low germination temperatures. For QPhs.ocs-4A.1, identified on the long arm of chromosome 4A, and QPhs.ocs-4B.1, on the centromeric region of the long arm of Chr 4B, the LOD peak positions and the desirable allele were consistent between the trials, while the LOD scores and contribution to the phenotypic variation varied. Transgressive segregants were observed among the 125 RILs and most of them had a combination of the three alleles conferring a higher dormancy: the Zen alleles at QPhs.ocs-3A.1 and QPhs.ocs-4A.1 and the CS allele at QPhs.ocs-4B1. This demonstrated a combined effect of the desirable alleles on accelerating grain dormancy, with their total effect being superior to that of Zen.     

Alteration of PHYA expression change circadian rhythms and timing of bud set in Populus

In many temperate woody species, dormancy is induced by short photoperiods. Earlier studies have shown that the photoreceptor phytochrome A (phyA) promotes growth. Specifically, Populus plants that over-express the oat PHYA gene (oatPHYAox) show daylength-independent growth and do not become dormant. However, we show that oatPHYAox plants could be induced to set bud and become cold hardy by exposure to a shorter, non-24 h diurnal cycle that significantly alters the relative position between endogenous rhythms and perceived light/dark cycles. Furthermore, we describe studies in which the expression of endogenous Populus tremula × P. tremuloides PHYTOCHROME A (PttPHYA) was reduced in Populus trees by antisense inhibition. The antisense plants showed altered photoperiodic requirements, resulting in earlier growth cessation and bud formation in response to daylength shortening, an effect that was explained by an altered innate period that leads to phase changes of clock-associated genes such as PttCO2. Moreover, gene expression studies following far-red light pulses show a phyA-mediated repression of PttLHY1 and an induction of PttFKF1 and PttFT. We conclude that the level of PttPHYA expression strongly influences seasonally regulated growth in Populus and is central to co-ordination between internal clock-regulated rhythms and external light/dark cycles through its dual effect on the pace of clock rhythms and in light signaling.     

Optimization of lentiviral vector transduction into peripheral blood mononuclear cells in combination with the fibronectin fragment CH-296 stimulation

Large scale T-cell expansion and efficient gene transduction are required for adoptive T-cell gene therapy. Based on our previous observations, human peripheral blood mononuclear cells (PBMCs) can be expanded efficiently while conserving a na?ve phenotype by stimulating with both recombinant human fibronectin fragment (CH-296) and anti-CD3 monoclonal antibodies. In this article, we explored the possibility of using this co-stimulation method to generate engineered T cells using lentiviral vector. Human PBMCs were stimulated with anti-CD3 together with immobilized CH-296 or anti-CD28 antibody as well as anti-CD3/anti-CD28 conjugated beads and transduced with lentiviral vector simultaneously. Co-stimulation with CH-296 gave superior transduction efficiency than with anti-CD28. Next, PBMCs were stimulated and transduced with anti-CD3/CH-296 or with anti-CD3/CD28 beads. T-cell expansion, gene transfer efficiencies and immunophenotypes were analysed. Stimulation with anti-CD3/CH-296 resulted in more than 10-times higher cell expansion and higher gene transfer efficiency with conservation of the na?ve phenotype compared with anti-CD3/CD28 stimulation method. Thus, lentiviral transduction with anti-CD3/CH-296 co-stimulation is an efficient way to generate large numbers of genetically modified T cells and may be suitable for many gene therapy protocols that use adoptive T-cell transfer therapy.     

Expression pattern of the CsPK3 auxin-responsive protein kinase gene

We have previously cloned a cDNA of a putative serine/threonine protein kinase gene named CsPK3 from cucumber, the mRNA level of which was up-regulated by auxin and down-regulated by light irradiation. To examine the CsPK3 gene expression in detail, we cloned a genomic DNA of CsPK3 gene and made transgenic tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) plants containing the fused CsPK3 promoter-beta-glucuronidase gene. The beta-glucuronidase expression was detected in the shoot apex, vascular tissues, and the outermost layer of cortex. The histological distribution of CsPK3 mRNA in cucumber seedlings was supported by in situ hybridization, where the positive signals were observed in similar tissues as those observed by beta-glucuronidase staining. The responsiveness of the CsPK3 gene to auxin and light was also confirmed for beta-glucuronidase activity. The pattern of beta-glucuronidase staining changed during the development of the tobacco seedlings. The results of our experiment showed that CsPK3 was expressed in a wide variety of tissues and cells in which the developmental and growth controls by auxin are suggested.