Responses to phosphate deprivation in white clover (Trifolium repens L.) : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy in Plant Biology at Massey University

Four breeding lines of white clover (Trifolium repens L.) were obtained from AgResearch Grasslands, Palmerston North, New Zealand, that had been shown previously to differ in terms of specific growth responses to added phosphate (P) in the field. These were designated Breeding Line (BL) 43 (low performer on low P; low performer on high P), BL 45 (low performer on low P; high performer on high P), BL 47 (high performer on low P; high performer on high P), and BL 49 (high performer on low P; low performer on high P). These breeding lines and five selected genotypes that were propagated from each line (designated 43-7, 43-8. 45-14. 45-4 and 47-9) were rooted in half-strength Hoagland solution in vermiculite for two weeks and then transferred to half-strength Hoagland liquid media for five weeks prior to the initiation of the experiments. For the breeding line screening, plants were acclimatized in a constant temperature environment for one week prior to treatments, while for the genotypic screening, plants were maintained in a temperature-controlled glasshouse. These lines and genotypes were characterized in relation to P uptake and utilization efficiency by growing in P-sufficient media (+P; 0.5 mM KH2PO4) and P-deficient media (-P; 0 mM KH2PO4) for 3, 5, 7 and 14 days (for the breeding line screening) and 7, 14 and 21 days (for the genotype screening). Over the time course, inorganic phosphate (Pi) content in leaves, non-specific acid phosphatase (APase) activity in intact roots (both as a total soluble activity and a cell-wall-associated activity), isoenzyme analyses, shoot dry weight (DW) and fresh weight (FW), leaf area, weight of an individual leaf (designated as the weight of the first fully expanded leaf), root FW, and the root:shoot (R:S) ratio were determined. Pi deprivation enhanced the induction of one major low mobility cell wall acidic isoform, two minor high mobility cell wall acidic isoforms and one major low mobility cell wall basic isoform in all genotypes. Furthermore, the activity of one major low mobility cell wall basic isoform was more higher in genotype 45-14 and one minor high mobility cell wall basic isoform was induced only in genotype 45-14 in response to Pi deprivation. In terms of individual BLs and genotypes, the screening results showed that BL 49 and genotype 45-14 displayed a constant Pi content and a slow induction of APase activity in the -P media, and had the highest total biomass FW in both +P and -P media. Overall (in both treatments) BL 49 and genotype 45-14 are the most efficient at utilizing available P as they produced the largest biomass FW, produced more roots in P- deprived media when compared with the other BLs and genotypes, and were more efficient in utilizing the P for the synthesis of biomass. BLs 43 and 45 and genotypes 43-7 and 43-8 are less efficient at utilizing available P, while under P deprivation, BL 45 and genotype 45-14 are the most efficient at utilizing P compared to the other BLs and genotypes. The study also showed that the Pi content in leaves and APase activity in roots was found to be the plant parameter most sensitive to Pi deprivation, and the results suggest that the selection of white clover germplasm for satisfactory performance under low P availability can be carried out using these two parameters as criteria

Effects of a functional rehabilitation program designed by the arronches care unit for a patient with stroke: a case report

This study describes the effect of a Functional Rehabilitation Program (FR) designed for the recovery of stroke patients. A new FR approach for stroke patients was experienced obeying the internment protocol used in the Continuing Care Units Mean Duration and Rehabilitation. According to the law (DC 101/2006), the protocol includes the patient initial assessment and the design of an individual FR program of treatment developed until 90 days; moreover, suggests evaluations after 30 days of hospitalization and before day 90 (Girão & Guerreiro, 2011). Case Description: Patient aged 74 yrs, diagnosed with left capsular hematoma striatum due to ischemic stroke. Background: hypertension and alcoholism. In the initial evaluation, the patient was oriented in time and space, he showed left hemiparesis, hearing and visual acuity impairment, motor programming deficit, balance at setting position compromised, inability to walk, and high dependence in basic and instrumentals activities of daily living (BADLs and IADLs). Evolution prognosis: reserved. Intervention: The FR program (5 days/week) incorporated three areas: Rehabilitation Nursing (RN), Physiotherapy (P), and Psychomotor Rehabilitation (PR).The RN performed BADLs relearning with a nurse; the P used the Bobath method; and the PR, as innovation, combined the traditional Brunnstrom method with sensorimotor stimulation targeting ontogenetic reliving. Evaluations were performed in the pre-intervention (PI), after 30 intervention days, and at the end of the program - before day 90. BADLs were accessed using the Barthel Index, and the IADLs using the Lawton Index. Global mobility was measured using Mobility Scale for the Elderly (EMI) and the six minutes walking test. The patient improved BADLs (Barthel, score: PI = 14; 30 days = 37; 90 days = 62) and IADLs (Lawton score: PI = 0; 30 days = 5; 90 day = 7), as well as mobility (EMI score: PI = 0; 30 days = 6; 90 day = 11), having reacquired the walking ability (walking test: PI = 0 m; 30 days = 38 m; 90 day = 50 m). The performed interdisciplinary intervention showed positive effects on the recovery of the stroke patient; specifically induced increases in his mobility and ability to perform BADLs and IADLs. These results suggest that interdisciplinary RF programs including ontogenetic reliving would be effective for the recovery of stroke patients with poor prognosis of evolution

Short-Term effect of COX-2 selective inhibitor as an adjunct for the treatment of periodontal disease: a clinical double-blind study in humans

Adjunctive therapeutic strategies that modulate the inflammatory mediators can play a significant role in periodontal therapy. In this double-blind, placebo-controlled study, 60 subjects diagnosed as periodontitis patients were evaluated for 28 days after periodontal treatment combined with selective cyclooxygenase-2 (COX-2) inhibitor. The experimental group received scaling and root planning (SRP) combined with the Loxoprofen antiinflammatory drug (SRP+Loxoprofen). The control group received SRP combined with placebo (SRP+placebo). Plaque index (PI), probing pocket depth (PD) and bleeding on probing (BOP) were monitored with an electronic probe at baseline and after 14 and 28 days. Both groups displayed clinical improvement in PD, PI and BOP. They also showed statistically similar values (p>0.05) of PD reduction on day 14 (0.4 mm) and on day 28 (0.6 mm). At the baseline, few deeper sites (>7 mm) from SRP+Loxoprofen group were responsible and most PD reduction was observed after 14 days (p<0.05). The percentage of remaining deep pockets (>7 mm) after 14 days in the SRP+Loxoprofen group was significantly lower (p<0.05) than in the SRP+placebo group. Loxoprofen presents potential effect as an adjunct of periodontal disease treatment, but long-term clinical trials are necessary to confirm its efficacy.Estratégias terapêuticas adjuvantes que modulam os mediadores inflamatórios podem ter função significante na terapia periodontal. Neste estudo duplo-cego controlado com placebo, 60 indivíduos diagnosticados com periodontite foram avaliados por 28 dias após tratamento periodontal combinado com inibidor seletivo de COX-2. O grupo experimental foi tratado com raspagem e alisamento radicular combinado com medicação anti-inflamatória Loxoprofeno (RAR+Loxoprofen). O grupo controle foi tratado com raspagem e alisamento radicular combinado com medicação placebo (Raspagem e alisamento radicular - RAR+placebo). Presença de placa (PI), profundidade de sondagem (PS) e sangramento à sondagem (SS) foram monitoradas com auxílio de uma sonda computadorizada no início do estudo e após 14 e 28 dias. Os dois grupos demonstraram melhora clínica em relação a PS, PI e SS. Também foi observado valores semelhantes (p>0,05) de redução da PS nos períodos de 14 dias (0,4 mm) e 28 dias (0,6 mm). No início do estudo, alguns sítios profundos (>7 mm) do grupo RAR+Loxoprofen foram os responsáveis pela maior redução da PS depois de 14 dias (p<0,05). A porcentagem de bolsas periodontais profundas >7 mm após 14 dias no grupo RAR+Loxoprofen foi significativamente inferior do que o grupo RAR+placebo (p<0.05). A medicação Loxoprofen apresenta potencial efeito adjuvante à terapia periodontal, mas estudos de longo prazo são necessários para confirmar sua eficácia.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)FAPESPSankyo Pharma Ltda

Lamivudine plus a boosted-protease inhibitor as simplification strategy in HIV-infected patients with toxicity to nucleoside analogues

Purpose of the study: Dual therapy with lamivudine plus a PI boosted with ritonavir (PI/r) could be an alternative to standard triple therapy or PI/r monotherapy as a simplification strategy in patients with toxicity to nucleoside analogues (NA). Methods: Retrospective cohort study of 44 HIV-infected patients on suppressive HAART, with no chronic HBV, who simplified to this dual therapy since 2008. Virological and immunological outcome, lipids and renal changes were evaluated. Summary of results: Mean age was 50 years (38-70), 66% were male, and the median time of HIV infection was 18.6 years. The median nadir CD4+ count was 150 cells/ml (2&#x2013;407). At inclusion, patients were receiving therapy with lamivudine plus atazanavir/r in 5 cases, lopinavir/r in 12, and darunavir/r in 27, and they had an HIV RNA level&#60;50 copies/ml for a median time of 794 days (129&#x2013;2344, 90% &#x003E;6 months). The NA discontinued was tenofovir (27), didanosine (12), AZT (3), and d4T (2). The reasons for changing were toxicity in 76% of cases, especially renal impairment. They had received a mean of 8 regimens before (2&#x2013;20), and 55% were in CDC-stage C. In 11 cases, history of resistance was available (to NA in 7 cases, including the 210W mutation in four). The mutations 184V was not observed, but four patients (9%) had a previous failure to therapy including 3tC. Mutations in the protease gene were observed in 8 patients (2 to 7 mutations, the most frequent 77I and 93L), without resistance to the current PI/r. During 62.8 patient-years of follow-up (median, 802 days), only 2 patients failed (4.5%), due to incomplete adherence, at 27 and 141 days. Of note, these two patients had no previous failed with 3tC or PI. Overall, CD4+ count increased for 55 cells/ml. No new adverse events were observed, but total cholesterol (from 180 to 246 mg/dl, p=0.007) and triglycerides (from 166 to 195, p=0.01) increased during the first 24 weeks with improvement at 48 weeks. On the other hand, estimated glomerular filtration rate improved during follow up (from 74.2 ml/min to 83.08 ml/min after 48 weeks, p=0.1). Conclusions: Dual therapy with lamivudine plus a boosted PI is safe and effective as simplification strategy in patients with toxicity to NA. This combination could be an alternative to mono or triple therapy in hard to treat patients, although an initial increase in lipid parameters could be observed

Atmospheric Charged K/πK/\pi Ratio and Measurement of Muon Annual Modulation with a Liquid Scintillation Detector at Soudan

We report a measurement of muon annual modulation in a 12-liter liquid scintillation detector with a live-time of more than 4 years at the Soudan Underground Laboratory. Muon minimum ionization in the detector is identified by its observed pulse shape and large energy deposition. The measured muon rate in the detector is 28.69$\pm$2.09 muons per day with a modulation amplitude of 2.66$\pm$ 1.0\% and a phase at Jul 22 $\pm$ 36.2 days. This annual modulation is correlated with the variation of the effective atmospheric temperature in the stratosphere. The correlation coefficient, $\alpha_{T}$, is determined to be $0.898 \pm 0.025$. This can be interpreted as a measurement of the atmospheric charged kaon to pion ($K$/$\pi$) ratio of 0.094$^{+0.044}_{-0.061}$ for $E_{p} >$ 7 TeV, consistent with the measurement from the MINOS far detector. To further constrain the value of $K$/$\pi$ ratio, a Geant4 simulation of the primary cosmic-ray protons with energy up to 100 TeV is implemented to study the correlation of $K$/$\pi$ ratio and the muon annual modulation for muon energy greater than 0.5 TeV. We find out that a charged $K$/$\pi$ ratio of 0.1598, greater than the upper bound (0.138) from this work at the production point 30 km above the Earth surface in the stratosphere cannot induce muon annual modulation at the depth of Soudan.Comment: 6 pages and 11 figure

Effect of Baking Soda-Peroxide Dentifrice on Post-Surgical Wound Healing

Purpose: To investigate the effect of a baking soda-hydrogen peroxide (0.75%) dentifrice on wound healing, plaque formation, gingival inflammation, wound healing, patient comfort, probing depth, and clinical attachment level following gingival flap surgery. Materials and Methods: A randomized, double-blind crossover study involving 25 patients requiring bilateral maxillary gingival flap surgery was completed. The effects of twice daily brushing with a baking soda-hydrogen peroxide dentifrice (Mentadent) or a placebo dentifrice (Crest) were observed over a 28-day post-surgical period. Gingival index (GI); plaque index (Pl) , probing depth (PD), clinical attachment level (CAL) and gingival bleeding index (Bl) were recorded pre-surgically and at day 28 for each surgical sextant. A.t days 7 and 14, soft tissue appearance/wound healing (STA) was assessed based on color and edema, PI\u27s were determined and patient comfort was ascertained by report. Results: Post-surgical wound healing was statistically significantly improved at day 7 with the trend continuing to day 14 when Mentadent dentifrice was used as compared to Crest dentifrice. However, there was no statistical difference in the PI values between the test and control dentifrice throughout the study. Use of Mentadent may be an effective aid in the early phase of healing following gingival flap surgery

Relationship between circulating levels of cortisol at slaughter and changes of some parameters of the camel meat during ageing

Relationship between serum levels of cortisol at slaughter and changes of some parameters of meat was investigated in 50 male camels, during ageing for 10 days at 4±1°C. Blood was sampled at slaughter to determine serum levels of cortisol and oblique abdominal external muscle was collected after slaughter to analyze pH, electrical conductivity (EC), drip loss (DL), cooked loss (CL), moisture, solids, ashes, protein total, calcium (Ca), phosphorus (Pi), magnesium (Mg), sodium (Na), potassium (K), catalase activiry (CATa), malondialdehyde (MDA), shrinkage, R-value and total haem pigment (THP) at days 0 (four hours postmortem), 5 and 10 of refrigerated storage. The camels were divided into 3 groups (Gr) with different serum levels of cortisol at slaughter (low, high and very high levels): Gr1 (13.07-67.9 ng/mL, n= 24), Gr2 (80.29-107.21 ng/mL, n= 7) and Gr3 (133.7-198.04 ng/mL, n= 19). Compared to Gr1, Gr2 and Gr3 had a significant (P<0.05) increase of pH, EC, DL, CL, THP, dimensional shrinkage and MDA, and a significant (P<0.05) decrease of CATa at the 5th and 10th postmortem days. However, the bleeding efficiency, moisture, ashes, R-value and levels of Ca, Pi, Na, K and total protein showed no significant differences between twoo cortisol groups. The antemortem circulating levels of cortisol were positively correlated with postmortem pH, EC, DL, CL, THP, dimensional shrinkage and MDA, and negatively correlated with CATa. The analysis of circulating cortisol at slaughtering may predict the quality and oxidant stress status of camel meat during postmortem cold storage

Immunomodulatory effects of seagrass Halophila ovalis polysaccharide mixed feed in adult black tiger shrimp Penaeus monodon and its protective efficacy against white spot syndrome virus infection

As white spot syndrome virus (WSSV) can be highly pathogenic in penaeid shrimp, various feed supplements have been tested to help to protect farmed shrimp against WSSV disease. Here a polysaccharide extract from Halophila ovalis (HO) seagrass was added to feeds at concentrations of 0.25, 0.5, and 1.0 g/kg to assess its ability to protect Black Tiger shrimp (Penaeus monodon) against WSSV challenge. Following feeding on these diets for 25 days, P. monodon were challenged by muscle injection and monitored for 21 days. On Day 0 and on Days 7 and 21 post-injection (pi), total haemocyte counts (THC), total protein concentrations, prophenoloxidase activity and respiratory burst activity were compared using haemolymph collected from 10 shrimp. All shrimp fed the basal diet died by Day 7 pi but survival times were extended among shrimp fed diets containing HO polysaccharide (HOP), and significantly at concentrations of 0.5 or 1 gkg^-1. Concomitantly with improved survival, all haemolymph immune parameters examined were enhanced significantly (p<0.05) among shrimp fed diets containing higher amounts of HOP. WSSV infection loads determined by real-time PCR were also lowered. The data suggest that if shrimp growth performance is not affected, inclusion of 0.5-1 gkg-1 HOP in commercial feeds might increase resilience of pond stocks of P. monodon against WSSV disease and when disease occurs, provide farmers with a longer management window to minimize economic losses